Degrader drug conjugates and methods of use thereof

Abstract

Conjugation reagents for making, and targeted degrader conjugates made therefrom, comprising a Janus family kinase binder and a CD127 binding agent, are described. In particular, the conjugation reagents are compounds of Formula (I) where TM is a JANUS family kinase domain-targeting binder; L1 is a first linker; L2 is a second linker; and E3L is a E3 ligase ligand; wherein, TM, L2 and E3L are covalently linked to L1, and L2 comprises a reactive moiety for reacting with a binding molecule. (I)

Description

DEGRADER DRUG CONJUGATES AND METHODS OF USE THEREOFCROSS-REFERENCE TO RELATED APPLICATION

[0001] The present application claims the benefit of priority of co-pending U.S.Provisional Patent Application No. 63 / 672,109, which was filed July 16, 2024, the content of which is incorporated herein by reference in its entirety.FIELD

[0002] The present disclosure relates to degrader payloads, degrader-drug conjugates and methods and uses thereof.INTRODUCTION

[0003] Degrader-drug conjugates and, more specifically, degrader antibody conjugates (DACs), are multifunctional molecules that comprise an antibody (delivery) component and a targeted protein degradation or proximity-induced protein degradation drug (payload) component, such as, for example, a heterobifunctional molecule comprising a target of interest binding component and an E3 ligase binding component. The drug targets a protein of interest by bringing together an E3 ligase binding component in close proximity to induce ubiquitination that tags the protein of interest for degradation (also known as a “kiss of death”) by hijacking of the cell’s ubiquitin proteasome degradation pathway to degrade the protein of interest. Certain types of molecular glues, that bring two molecules together that do not normally interact, could also be used as a payload for proximity-induced protein degradation.

[0004] There is interest in developing precision immunology treatments for immunological disorders to selectively target cell types and pathways in those cell types that initiate or propagate disease, while sparing certain cell types or avoiding activating these pathways in other cells, thus maximizing therapeutic benefit while minimizing side-effects and toxicities. The JAK / STAT signaling pathways play a multitude of roles in immunology in the transduction of key cytokines signals in a variety of cells. T argeting and inhibiting the activity of Janus family of kinases (JAK1 / JAK2 / JAK3 / TYK2) is an approach being assessed for inhibiting a variety of autoimmune disorders. While a number of pan-JAK inhibitors and inhibitors with increased specificity for one or more of the kinases in this family are approved or in clinical development for the treatment of various autoimmune disorders, the broad acting small molecules are acting on a variety of cells and this results in a broad range of toxicities limiting dosing or their use in broader indications and earlier lines of therapy, including black box FDA warnings on the label of JAK1-3-targeting molecules and increased risks of SAEs such as malignancies, MACEs, thromboembolism, serious infections and all-cause mortality. [PMID: 38435420] While mechanisms for the above serious adverse effects are not fully understood,it is clear that the activity of JAK-family modulating compounds on this pathway in certain cell types is driving the unwanted side effects. For example, occurrence of serious thromboembolic events is likely a result of JAK-family inhibitors acting on prothrombotic pathways and mechanisms, such as endothelial activation, platelet aggregation, neutrophils and NETs, and possibly expression of tissue factor and adhesion molecules [PM ID: 38435420], More selective inhibition of the JAKs, for example, in certain T cells in immune disorders where these cells are known to drive disease progression, would avoid inhibiting JAKs in cells that may drive the occurrence of JAK-family inhibitor-associated adverse effects.

[0005] The mechanisms of autoimmune disease are complex and incompletely understood.

[0006] US Patent 9,315,494B2 is directed to alkyl-amide-substituted pyridyl compounds useful as modulators of IL-12, IL-23 and / or IFNa responses via Tyk2. US Patent 11 ,053,219 B2 is directed to substituted pyridine TYK2 inhibitors.

[0007] W02022100710A1 discloses Tyrosine kinase 2 (TYK2) degradation compounds and methods of use. WO2023076161A1 is directed to Tyk2 degraders and uses thereof and WO 2024 / 020221 A1 is directed to modulators of Tyk2 proteolysis and associated methods of use. WO 2023 / 054549 A1 is directed to a degradation inducer and WO 2021 / 141662 A1 is directed to proteolysis targeting chimeric molecules (PROTACs) with functional handles and uses thereof. WO 2022 / 093742A1 is directed to compounds for targeted protein degradation of kinases.

[0008] Small molecule inhibitors, modulators and degraders of Tyk2 (and other Jak- family members) can lack cell type- or subtype-specific context, i.e. they are untargeted and will modulate the JAK / STAT pathways to varying degrees in a broad variety of cell types in the body, where these pathways all drive different functions. Aside from the side-effects associated with this untargeted approach, one may also not fully realize the efficacy potential of the inhibition of this pathway to treat immune disorders because of dosing limitations.

[0009] A need for additional modalities to treat disorders such as autoimmune diseases (IBD, UC, CD, PsO, PsA, AD, SLE, T1 DM and others) and disorders with an autoimmune component (such as T2DM) remains.SUMMARY

[0010] The present disclosure provides in an aspect, heterobifunctional compounds including conjugation reagents and degrader-antibody conjugates (DACs) for targeted delivery of a degrader payload. Such compounds can for example be used for the treatment of immunological disorders, such as autoimmune disorders. The mechanisms of autoimmune disease are complex and incompletely understood, yet certain subsets of cells are known toplay an increased or particular role in the initiation or progression of various autoimmune disorders. T cells play an important role in a number of autoimmune conditions, and targeting the abnormal activity of specific subsets of T cells and not others may present an opportunity for a more precise strategy for amelioration of these conditions, while minimizing treatment- induced side effects. Disclosed herein are specific proximity-induced protein degraders of JAK1 and / or TYK2 kinases (via the targeting of for example their JH2 pseudokinase domains) targeted via CD127 (as, for example, an antibody-conjugated payload) to Th (T helper), T effector and central memory cells, and Tfh (T follicular helper) cells, while optionally also sparing the beneficial Treg (T regulatory) cells. More specifically, disclosed are specific proximity-induced single and dual protein degraders of TYK2 and TYK2 / JAK1, respectively, targeted to CD127 (as, for example, an antibody-drug conjugate).

[0011] These and other features and advantages of the present disclosure will become apparent from the following detailed description taken together with the accompanying drawings. It should be understood, however, that the detailed description and specific examples, while indicating preferred implementations of the present disclosure, are given by way of illustration only, since various changes and modifications within the spirit and scope of the disclosure will become apparent to those of skill in the art from this detailed description.BRIEF DESCRIPTION OF THE DRAWINGS

[0012] For a better understanding of the various embodiments described herein, and to show more clearly how these various embodiments may be carried into effect, reference will be made, by way of example, to the accompanying drawings which show at least one example embodiment, and which are now described. The drawings are not intended to limit the scope of the teachings described herein.

[0013] Fig. 1 Ato 1 E are a series of experiments showing CD127 expression of various in vitro cell models, where Fig. 1A is a flow cytometry density plot of Peripheral Blood Mononuclear Cells (PMBCs) stained with (right plot) anti-CD3-BV605 and anti-CD127-APC antibodies or control (left plot); Fig. 1 B is a flow cytometry histogram plot of Peripheral Blood Mononuclear Cells (PMBCs) stained with anti-CD127-APC antibody; Fig. 1C is a flow cytometry histogram plot of various in vitro cell models (PMBCs, Molt-4, Jurkat E6.1) stained with anti- CD127-APC antibody, Fig. 1D is a graph of CD127 expression of various in vitro cell models (Jurkat E6.1 ); and Fig. 1 E is a graph of CD127 expression of various in vitro cell models (Molt- 4).

[0014] Fig. 2 is a table of set-up of pHrodo dye-labelled anti-CD127 antibody internalization IncuCyte experiment in Jurkat, Molt4 and PBMC cells (100,000 cells per well).

[0015] Fig. 3A to 3F are a series of images showing internalization of Zenon-red- labelled (pHrodo iFL Red for human IgG labelling) anti-CD127 antibodies by PBMCs, where arrows indicate bright red fluorescence, where in Fig. 3A anti-CD127 antibodies are GNP, in Fig. 3B anti-CD127 antibodies are GDS, in Fig. 3C anti-CD127 antibodies are PHP, in Fig. 3D anti-CD127 antibodies are PHS, in Fig. 3E anti-CD127 antibodies are MOP, in Fig. 3F anti- CD127 antibodies are MPH.

[0016] Fig. 4 is a graph of Zenon red labelled anti-CD127 antibody internalization kinetics in PBMCs. Counts represent cells internalizing significant amounts of antibody to reach pH-dependent fluorescence signal threshold.

[0017] Fig. 5A to 5F are a series of images showing internalization of pHrodo-green direct labelled anti-CD127 antibodies by PBMCs, where arrows indicate bright green fluorescence, where in Fig. 5A anti-CD127 antibodies are GNP, in Fig. 5B anti-CD127 antibodies are GDS, in Fig. 5C anti-CD127 antibodies are PHP, in Fig. 5D anti-CD127 antibodies are PHS, in Fig. 5E anti-CD127 antibodies are MCP, in Fig. 5F anti-CD127 antibodies are MPH.

[0018] Fig. 6 is an image of Western blot analysis of the degradation of Jak1 , Tyk2 and phosphorylation of downstream Statl and Stat3 in Jurkat cells treated with compounds I- 5, I-6, I-7, I-8, I-9 and 1-10 (with quenched sulfhydryl-reactive moieties).

[0019] Fig. 7 is an image of Western blot analysis of the degradation of Jak1 and Tyk2 and phosphorylation of downstream Statl and Stat 3 in PBMCs treated with PHP anti-CD127 Ab DACs with compounds I-5, I-6, I-7, I-8, I-9 and 1-10.

[0020] Fig. 8 is an image showing an example of a model of a degrader payload cleaved off an antibody and complexed to induce the “Kiss of Death” between Cereblon E3 ligase and Tyk2.

[0021] Fig. 9 is an image of Western blot showing expression and activation levels ofJAK1 , TYK2, STAT1 and STAT3 in lysates of human CD127+ T cells sorted from human PBMCs and treated with various DACs (analyzed at 72 hours post-treatment). (GH = GNP anti- CD127 Ab DAC with compound 1-10; PH = PHP anti-CD127 Ab DAC with compound 1-10, PF = PHP anti-CD127 Ab DAC with compound I-6, PG = PHP anti-CD127 Ab DAC with compound I-9, GE = GNP anti-CD127 Ab DAC with compound I-5).

[0022] Fig. 10 is a graph of % weight changes in T cell transfer colitis animal model of IBD, with groups showing healthy controls (no donor cells infused), vehicle treated animals (PBS control) and DAC treated animals (GDS anti-CD127 Ab DAC with compound 1-21 ; early treatment). Approx. 0.5x106Donor CD4+ CD45RBhiahT cells from C57BL / 6-ll7rTM1(IL7R) ECDtransgenic mice were transferred into RAG1 KO recipients to generate a model of human IBD in mice, which were monitored for changes in weight and fecal lipocalin-2. Animals were treated as indicated, and additional markers of inflammation and cells were analyzed on Day 70.

[0023] Fig. 11 is a graph of changes in fecal lipocalin-2 in T cell transfer colitis animal model of IBD, with groups showing healthy controls (no donor cells infused), vehicle treated animals (PBS control) and DAC treated animals (early treatment). Approx. 0.5x10® Donor CD4+ CD45RBh'9hT cells from C57BL / 6-ll7rTM1(IL7R) ECD transgenic mice were transferred into RAG1 KO recipients to generate a model of human IBD in mice, which were monitored for changes in weight and fecal lipocalin-2. Animals were treated as indicated, and additional markers of inflammation and cells were analyzed on Day 70.

[0024] Fig. 12 is a graph of % weight changes in T cell transfer colitis animal model of IBD, with groups showing healthy controls (no donor cells infused), vehicle treated animals (PBS control), DAC treated animals (GDS anti-CD127 Ab DAC with compound 1-21 ; late treatment of severe disease). Approx. 0.5x10® Donor CD4+ CD45RBh'9hT cells from C57BL / 6- H7rTM1 (IL7R) ECD transgenic mice were transferred into RAG1 KO recipients to generate a model of human IBD in mice, which were monitored for changes in weight and fecal lipocalin- 2. Animals were treated as indicated, and additional markers of inflammation and cells were analyzed on Day 70.

[0025] Fig. 13 is a graph of % weight changes in T cell transfer colitis animal model of IBD, with groups showing healthy controls (no donor cells infused), vehicle treated animals (PBS control) and upadacitinib treated animals (JAK inhibitor control). Approx. 0.5x10® Donor CD4+ CD45RBh'9hT cells from C57BL / 6-ll7rTM1(IL7R) ECD transgenic mice were transferred into RAG1 KO recipients to generate a model of human IBD in mice, which were monitored for changes in weight and fecal lipocalin-2. Animals were treated as indicated, and additional markers of inflammation and cells were analyzed on Day 70.

[0026] Fig. 14 is a graph of changes in fecal lipocalin-2 in T cell transfer colitis animal model of IBD, with groups showing healthy controls (no donor cells infused), vehicle treated animals (PBS control), DAC treated animals (late treatment of severe disease). Approx. 0.5x10® Donor CD4+ CD45RBh'9hT cells from C57BL / 6-ll7rTM1(IL7R) ECD transgenic mice were transferred into RAG1 KO recipients to generate a model of human IBD in mice, which were monitored for changes in weight and fecal lipocalin-2. Animals were treated as indicated, and additional markers of inflammation and cells were analyzed on Day 70.

[0027] Fig. 15 is a graph of fold changes in fecal lipocalin-2 post-treatment in T cell transfer colitis animal model of IBD, with groups showing vehicle treated animals (PBS control), DAC treated animals (late treatment of severe disease) and upadacitinib treated animals (JAKinhibitor control). Approx. 0.5x10® Donor CD4+ CD45RBhiahT cells from C57BL / 6-ll7rTM1(IL7R) ECD transgenic mice were transferred into RAG1 KO recipients to generate a model of human IBD in mice, which were monitored for changes in weight and fecal lipocalin-2. Animals were treated as indicated, and additional markers of inflammation and cells were analyzed on Day 70.

[0028] Fig. 16 is a representative flow cytometry analysis of the CD127+ donor- derived CD4 T cell populations in T cell transfer colitis animal model of IBD, with groups showing vehicle treated animals (PBS control) and DAC treated animals (late treatment of severe disease). Approx. 0.5x10® Donor CD4+ CD45RBh'9hT cells from C57BL / 6-ll7rTM1(IL7R) ECD transgenic mice were transferred into RAG1 KO recipients to generate a model of human IBD in mice, which were monitored for changes in weight and fecal lipocalin-2. Animals were treated as indicated, and additional markers of inflammation and cells were analyzed on Day 70.

[0029] Fig. 17 is a summary of flow cytometry analysis of changes in the CD127+ donor-derived T cell colonic and splenic populations in T cell transfer colitis animal model of IBD, with groups showing vehicle treated animals (PBS control), DAC treated animals (late treatment of severe disease) and upadacitinib treated animals (JAK inhibitor control). Approx. 0.5x10® Donor CD4+ CD45RBhigh T cells from C57BL / 6-ll7rTM1(IL7R) ECD transgenic mice were transferred into RAG1 KO recipients to generate a model of human IBD in mice, which were monitored for changes in weight and fecal lipocalin-2. Animals were treated as indicated, and additional markers of inflammation and cells were analyzed on Day 70.

[0030] Fig. 18 is an image of representative colon histological sections capturing elements of disease progression and inflammation in T cell transfer colitis animal model of IBD, with groups showing healthy animals (no donor cells infusion), vehicle (PBS) treated animals, DAC treated animals and upadacitinib treated animals. Approx. 0.5x10® Donor CD4+ CD45RBhigh T cells from C57BL / 6-ll7rTM1(IL7R) ECD transgenic mice were transferred into RAG1 KO recipients to generate a model of human IBD in mice. Animals were treated as indicated, and sections of colon were obtained on Day 70 and processed for histology. H&E sections of representative colon sections are shown at 10x magnification.DESCRIPTION OF VARIOUS EMBODIMENTS

[0031] Unless otherwise indicated, the definitions and embodiments described in this and other sections are intended to be applicable to all embodiments and aspects of the present disclosure herein described for which they are suitable as would be understood by a person skilled in the art.

[0032] All features disclosed in the specification, including the claims, abstract, and drawings, and all the steps in any method or process disclosed, may be combined in any combination, except combinations where at least some of such features and / or steps are mutually exclusive. Each feature disclosed in the specification, including the claims, abstract, and drawings, can be replaced by alternative features serving the same, equivalent, or similar purpose, unless expressly stated otherwise.I. Definitions

[0033] In understanding the scope of the present disclosure, the term "comprising" and its derivatives, as used herein, are intended to be open ended terms that specify the presence of the stated features, elements, components, groups, integers, and / or steps, but do not exclude the presence of other unstated features, elements, components, groups, integers and / or steps. The foregoing also applies to words having similar meanings such as the terms, "including", "having" and their derivatives.

[0034] The term “consisting” and its derivatives, as used herein, are intended to be closed ended terms that specify the presence of stated features, elements, components, groups, integers, and / or steps, and also exclude the presence of other unstated features, elements, components, groups, integers and / or steps. The term “consisting essentially of”, as used herein, is intended to specify the presence of the stated features, elements, components, groups, integers, and / or steps as well as those that do not materially affect the basic and novel characteristic(s) of features, elements, components, groups, integers, and / or steps.

[0035] Further, terms of degree such as "substantially", "about" and "approximately" as used herein mean a reasonable amount of deviation of the modified term such that the end result is not significantly changed. These terms of degree should be construed as including a deviation of at least ±5% of the modified term if this deviation would not negate the meaning of the word it modifies.

[0036] The terms "about", “substantially” and “approximately” as used herein mean a reasonable amount of deviation of the modified term such that the end result is not significantly changed. These terms of degree should be construed as including a deviation of at least ±5% of the modified term if this deviation would not negate the meaning of the word it modifies or unless the context suggests otherwise to a person skilled in the art.

[0037] As used in this specification and the appended claims, the singular forms “a”,“an” and “the” include plural references unless the content clearly dictates otherwise. Thus, for example, a composition containing “a compound” includes a mixture of two or more compounds.

[0038] The phrase "and / or," as used herein in the specification and in the claims, should be understood to mean "either or both" of the elements so conjoined, i.e. , elements that are conjunctively present in some cases and disjunctively present in other cases. Multiple elements listed with "and / or" should be construed in the same fashion, i.e., "one or more" of the elements so conjoined. Other elements may optionally be present other than the elements specifically identified by the "and / or" clause, whether related or unrelated to those elements specifically identified.

[0039] As used herein, "or" should be understood to have the same meaning as"and / or" as defined above. For example, when separating items in a list, "or" or "and / or" shall be interpreted as being inclusive, i.e., the inclusion of at least one, but also including more than one, of a number or list of elements, and, optionally, additional unlisted items. Only terms clearly indicated to the contrary, such as "only one of or "exactly one of' or, when used in the claims, "consisting of' will refer to the inclusion of exactly one element of a number or list of elements. In general, the term "or" as used herein shall only be interpreted as indicating exclusive alternatives (i.e., "one or the other but not both") when preceded by terms of exclusivity, such as "either," "one of," "only one of," or "exactly one of."

[0040] As used herein, the phrase "at least one," in reference to a list of one or more elements, should be understood to mean at least one element selected from anyone or more of the elements in the list of elements, but not necessarily including at least one of each and every element specifically listed within the list of elements and not excluding any combinations of elements in the list of elements. This definition also allows that elements may optionally be present other than the elements specifically identified within the list of elements to which the phrase "at least one" refers, whether related or unrelated to those elements specifically identified.

[0041] The recitation of numerical ranges by endpoints herein includes all numbers and fractions subsumed within that range (e.g., 1 to 5 includes 1 , 1.5, 2, 2.75, 3, 3.90, 4, and 5). It is also to be understood that all numbers and fractions thereof are presumed to be modified by the term "about."

[0042] The term “target moiety (TM)” and its derivatives, as used herein, refers to an agent that specifically interacts with (e.g. binds with) a target such as Tyk2 / Jak1 and which may also directly reduce, decrease, or otherwise block expression or activity of the target, and includes any substance that is capable of inhibiting the expression or activity of the target and includes, without limitation, small molecules, aptamers, proteins, peptide mimetics, antibodies (and fragments thereof), and other substances that specifically bind the target. For example,the TM can be a JH2 pseudokinase domain-targeting binder, optionally an inhibitor, targeting TYK2 and / or JAK1.

[0043] The term “cell” as used herein refers to a single cell or a plurality of cells and includes a cell either in a cell culture or in a subject.

[0044] The term “amino acid” includes all of the naturally occurring amino acids as well as modified L- and D- amino acids. The atoms of the amino acid can for example include different isotopes. For example, the amino acids can comprise deuterium substituted for hydrogen, nitrogen-15 substituted for nitrogen-14, and carbon-13 substituted for carbon-12 and other similar changes.

[0045] The term "antibody" as used herein is intended to include monoclonal antibodies, polyclonal antibodies, single chain antibodies, chimeric and humanized antibodies and binding fragments thereof, including for example a single chain Fab fragment, Fab’2 fragment, or single chain Fv fragment as well as multispecific (e.g. bispecific) antibodies. The antibody may be from recombinant sources and / or produced in transgenic animals. Also included are human antibodies or binding fragments thereof that can be produced in transgenic animals or using biochemical techniques, or can be isolated from a library such as a phage display library or a Fab expression library. Humanized or other chimeric antibodies may include sequences from one or more than one isotype, class, or species. Antibodies may be any class of immunoglobulins including: IgG, IgM, IgD, IgA, or IgE; and any isotype thereof, including lgG1 , lgG2 (e.g. lgG2a, lgG2b), lgG3 and lgG4. Further, these antibodies are typically produced as antigen binding fragments such as Fab, Fab' F(ab')2, Fd, Fv and single domain antibody fragments, or as single chain antibodies in which the heavy and light chains are linked by a spacer. The antibodies may include sequences from any suitable species including human. Also, the antibodies may exist in monomeric or polymeric form.

[0046] The term "antibody fragment" or “binding fragment” as used herein is intended to include without limitations Fab, Fab', F(ab')2, scFab, scFv, dsFv, ds-scFv, Fc-fusion proteins, dimers, minibodies, diabodies, and multimers thereof, multispecific antibody fragments and Domain Antibodies. Antibodies can be fragmented using conventional techniques. For example, F(ab')2 fragments can be generated by treating the antibody with pepsin. The resulting F(ab')2 fragment can be treated to reduce disulfide bridges to produce Fab' fragments. Papain digestion can lead to the formation of Fab fragments. Fab, Fab' and F(ab')2, scFv, dsFv, ds-scFv, Fc-fusion proteins, dimers, minibodies, diabodies, bispecific antibody fragments and other fragments can also be synthesized by recombinant techniques.

[0047] The term “complementarity determining region” or “CDR” as used herein refers to particular hypervariable regions of antibodies that are involved in epitope binding. Thegeneral structure and properties of CDRs of antibodies have been described in the art. Briefly, in an antibody scaffold, the CDRs are embedded within a framework in the heavy and light chain variable region where they constitute the regions largely responsible for antigen binding and recognition. A variable region typically comprises at least three heavy or light chain CDRs (Kabat et al., 1991, Sequences of Proteins of Immunological Interest, Public Health Service N.I.H., Bethesda, Md.; see also Chothia and Lesk, 1987, J. Mol. Biol. 196:901 -917; Chothia et al., 1989, Nature 342: 877-883), within a framework region (designated framework regions 1- 4, FR1, FR2, FR3, and FR4, by Kabat et al., 1991 ; see also Chothia and Lesk, 1987, supra). CDRs can be annotated in various ways including the method according to Kabat, AbM, or IMGT. Accordingly, the CDRs of the same antibody can comprise different sequences, depending on which method was used to annotate the CDR sequences. Computational methods can be used for identifying CDR sequences include Kabat, Chothia, IgBlast and IMGT. The CDRs listed in the present disclosure are identified using the Kabat system. A person skilled in the art having regard to the sequences comprised herein would also be able to identify CDR sequences based on IMGT, Kabat and Chothia etc.

[0048] The term “affinity", as used herein, refers to the strength of the sum total of noncovalent interactions between a single binding site of a molecule and its binding partner. Unless indicated otherwise, as used herein, "binding affinity" refers to intrinsic binding affinity which reflects a 1 :1 interaction between members of a binding pair. The affinity of a molecule X for its partner Y can generally be represented by the equilibrium dissociation constant (KD). Affinity can be measured by common methods known in the art.

[0049] Generally, reference to a "peptide” includes reference to a peptide, polypeptide or protein or parts thereof. The peptide may be glycosylated or non-glycosylated, and / or may contain a range of other molecules fused, linked, bound or otherwise associated to the protein such as amino acids, lipids, carbohydrates or other peptides, polypeptides or proteins. Reference hereinafter to a "peptide" includes a peptide comprising a sequence of amino acids as well as a peptide associated with other molecules such as amino acids, lipids, carbohydrates or other peptides, polypeptides or proteins.

[0050] The term “Janus family kinase” or “JAKs”, as used herein refer to a family of non-receptor tyrosine kinases, including Jak1 , Jak2, Jak3 and tyrosine kinase 2 (Tyk2). JAKs facilitate signal transduction by phosphorylating the relevant receptor

[0051] The term “Ubiquitin ligase ligand” refers to a compound that binds ubiquitin ligase, such as an E3 ligase (“E3L”) which includes E3 ligase and E3 ligase complex components. In heterobifunctional molecules comprising such a ligand, and a moiety that binds a target protein, binding of ubiquitin and the target results via induced proximity, in ubiquitinationand degradation of the target protein or molecule. Examples include without limitation, E3 ligase ligands, such as von Hippel-Landau (VHL) ligands or cereblon (CRBN) ligands including for example, thalidomide, lenalidomide, pomalidomide and derivatives with an N-alkylated glutarimide ring and derivatives thereof that bind E3 ligase with a minimum binding affinity of at least 100 nM, for example about 100 nM to 200 nM, or more than 200 nM.

[0052] The term “CD127” also referred to as “IL-7Ra”, “IL-7RA” and “CD127 / IL-7Ra”, as known in the art is a 75-80 kDa glycoprotein that can form a heterodimer with the common- y-chain receptor. CD127 is expressed in various T cell compartments and can be upregulated on activated T-helper (Th) cells, T follicular helper (Tfh) cells, activated T cells, T effector memory cells (Tern), T central memory cells (Tern), and downregulated on T regulatory (Treg) cells. CD127 includes known sequences such as the nucleotide and amino acid sequences of CD127, including CD127 isoforms, and particularly human CD127, the sequence of which can be found, for example at UniProt ID: P16871 , or GenBank Accession No: NP_002176.

[0053] The term "subject" as used herein includes all members of the animal kingdom including mammals, and suitably refers to humans. Optionally, the term “subject” includes mammals that have been diagnosed with an autoimmune disease. In one embodiment, the term “subject” refers to a human having, or suspected of having, an autoimmune disease.

[0054] The term "subject in need thereof' refers to a subject that could benefit from the method(s) or treatment(s) described herein, and optionally refers to a subject with an autoimmune disease, or optionally a subject with increased risk of an autoimmune disease, such as a subject with a strong genetic disposition.

[0055] The term “autoimmune disease” as used to herein includes diseases which have an autoimmune component. Autoimmune diseases include rheumatoid arthritis (RA), systemic lupus erythematosus (SLE), celiac disease (CD), inflammatory bowel disease (IBD), ulcerative colitis (UC), psoriasis (PsO), psoriatic arthritis (PsA), atopic dermatitis (AD), diabetes, including T1 D, T2D, immune checkpoint inhibitor-induced type 1 diabetes. The autoimmune diseases can include upregulation of CD127 in one or more T cell compartments.

[0056] The term “CD127 mediated disorder” includes diseases where cells that express CD127 (CD127-positive) are involved in the mechanism of disease, such as autoimmune diseases, and either initiate, drive, exacerbate, and / or maintain disease.

[0057] The term "treating" or “treatment” as used herein and as is well understood in the art, means an approach for obtaining beneficial or desired results, including clinical results. Beneficial or desired clinical results can include, but are not limited to, alleviation or amelioration of one or more symptoms or conditions, diminishment of extent of disease, stabilized (i.e. , not worsening) state of disease, delay or slowing of disease progression, amelioration of thedisease state, diminishment of the reoccurrence of disease, and remission (whether partial or total).

[0058] The term "administered" or “administering” as used herein means administration of a therapeutically effective amount of a compound or composition of the disclosure to a subject or a cell for example in cell culture or in a patient.

[0059] As used herein, the phrase "effective amount" or "therapeutically effective amount" means an amount effective, at dosages and for periods of time necessary to achieve the desired result. For example, in the context of treating an autoimmune disease, an effective amount is an amount that for example induces remission, reduces disease severity, and / or slows disease progression compared to the response obtained without administration of the compound.

[0060] The term “pharmaceutically acceptable” means compatible with the treatment of subjects, for example humans.

[0061] The term “pharmaceutically acceptable carrier” means a non-toxic solvent, dispersant, excipient, adjuvant or other material which is mixed with the active ingredient in order to permit the formation of a pharmaceutical composition, i.e., a dosage form capable of administration to a subject.

[0062] The term “parenteral” as used herein includes percutaneous, subcutaneous, intravascular (e.g., intravenous), or intramuscular, injection or infusion techniques and the like.

[0063] The term “protecting group” or “PG” and the like as used herein refers to a chemical moiety which protects or masks a reactive portion of a molecule to prevent side reactions in those reactive portions of the molecule, while manipulating or reacting a different portion of the molecule. After the manipulation or reaction is complete, the protecting group is removed under conditions that do not degrade or decompose the remaining portions of the molecule. The selection of a suitable protecting group can be made by a person skilled in the art. Many conventional protecting groups are known in the art, for example as described in “Protective Groups in Organic Chemistry” McOmie, J.F.W. Ed., Plenum Press, 1973, in Greene, T.W. and Wuts, P.G.M., “Protective Groups in Organic Synthesis”, John Wiley & Sons, 3rdEdition, 1999 and in Kocienski, P. Protecting Groups, 3rd Edition, 2003, Georg Thieme Verlag (The Americas).

[0064] The term “inert organic solvent” as used herein refers to a solvent that is generally considered as non-reactive with the functional groups that are present in the compounds to be combined together in any given reaction so that it does not interfere with orinhibit the desired synthetic transformation. Organic solvents are typically non-polar and dissolve compounds that are non-soluble in aqueous solutions.

[0065] The term “alkyl” as used herein, whether it is used alone or as part of another group, means straight or branched chain, saturated alkyl groups. The number of carbon atoms that are possible in the referenced alkyl group are indicated by the prefix “Cni-n2”. For example, the term Ci-ioalkyl means an alkyl group having 1 , 2, 3, 4, 5, 6, 7, 8, 9 or 10 carbon atoms.

[0066] The terms “halo” or “halogen” as used herein, whether it is used alone or as part of another group, refers to a halogen atom and includes fluoro, chloro, bromo and iodo.

[0067] The term “cross-coupling” as used herein refers to chemical reactions in which two different starting materials, each of which is usually endowed with an activating group, are reacted together with the aid of a metal catalyst. The result is the loss of the two activating groups and the formation of a new covalent bond between the remaining fragments.

[0068] The term “solvate” as used herein means a compound, or a salt and / or prodrug of a compound, wherein molecules of a suitable solvent are incorporated in the crystal lattice. A suitable solvent is physiologically tolerable at the dosage administered.

[0069] The term “prodrug” as used herein means a compound, or salt and / or solvate of a compound, that, after administration, is converted into an active drug.

[0070] The term “linker” as used herein refers to any molecular structure that joins two or more other molecular structures together.

[0071] The term “reacts with” as used herein generally means that there is a flow of electrons or a transfer of electrostatic charge resulting in the formation of a covalent bond.

[0072] The term “conjugating” as used herein means to bind two molecules together via a covalent bond.

[0073] The present description refers to a number of chemical terms and abbreviations used by those skilled in the art. Nevertheless, definitions of selected terms are provided for clarity and consistency.

[0074] The term “Me” as used herein refers to methyl.

[0075] The term “Et” as used herein refers to ethyl.

[0076] The term “Pr” as used herein refers to propyl.

[0077] The term “Bu” as used herein refers to butyl.

[0078] The term “Ac” as used herein refers to acetyl.

[0079] The term “Ph” as used herein refers to phenyl.

[0080] The term “Boo” as used herein refers to tert-butyloxycarbonyl.

[0081] The term “Fmoc” as used herein refers to fluorenylmethoxy carbonyl

[0082] The term “Bpin” as used herein refers to pinacol boronate

[0083] The terms “SPDP” as used herein refer to N-Succinimidyl 4-(2- pyridyldithio)butanoate

[0084] The term “sulfo” as used herein refer to sulfonic acid group.

[0085] The terms “DM AC” as used herein refer to diacetamide group.

[0086] The term “SPP” as used herein refer to N-succinimidyl 4-(2- pyridyldithio)pentanoate.

[0087] The term “SPDMV” as used herein refer to 2,5-Dioxopy rrolidin-1 -yl 4-methyl-4-(pyridin-2-yldisulfaneyl)pentanoate.

[0088] The term “Py” as used herein refer to pyridyl group.

[0089] The term “ds” or “SS” as used herein refer to disulfanyl group.

[0090] The term “Pfp” as used herein refer to pentafluorophenyl group.

[0091] The term “PEG” as used herein refer to polyethylene glycol.

[0092] The term “NHS” as used herein refer to N-hydroxysuccinimide.

[0093] The term “PAB” as used herein refer to para-aminobenzyl alcohol.

[0094] The term “PNP” as used herein refer to para-nitrophenyl.

[0095] The term “Mai” as used herein refer to maleimide.

[0096] The term “MP” as used herein refer to maleimidopropionyl.

[0097] The term “MC” as used herein refer to maleimidocaproyl.

[0098] The term “Ab” as used herein refer to antibody.

[0099] It should also be understood that, in certain methods described herein that include more than one step or act, the order of the steps or acts of the method is not necessarily limited to the order in which the steps or acts of the method are recited unless the context indicates otherwise.

[0100] Further, the definitions and embodiments described in particular sections are intended to be applicable to other embodiments herein described for which they are suitable aswould be understood by a person skilled in the art. For example, in the following passages, different aspects are defined in more detail. Each aspect so defined may be combined with any other aspect or aspects unless clearly indicated to the contrary. In particular, any feature indicated as being preferred or advantageous may be combined with any other feature or features indicated as being preferred or advantageous.

[0101] Although any methods and materials similar or equivalent to those described herein can also be used in the practice or testing of the present disclosure, examples of methods and materials are now described.II. Conjugation reagents

[0102] In one aspect of the disclosure, there is provided a conjugation reagent according to the formula (I):wherein TM is a Janus family kinase binder; L1is a linker; L2is a binding molecule linker; and E3L is an E3 ligase ligand; wherein, ITM, L2and E3L are covalently linked to L1and L2has a reactive moiety for reacting with a binding molecule.

[0103] In some embodiments, the TM is a Janus family kinase binder that targets the JH2 pseudokinase domain (e.g. with or without inhibiting kinase function). In other embodiments, the TM targets JH1 catalytic domain. The TM in addition to being a Janus family kinase binder, can also be a Janus family kinase inhibitor that inhibits one or more members of the Janus family.

[0104] In some embodiments, the TM is a Janus family kinase inhibitor that targets the JH2 pseudokinase domain. In additional embodiments, the TM is a Janus family kinase inhibitor that targets the JH1 catalytic domain.

[0105] In some embodiments, the Janus family kinase binder is a JAK1 , JAK2, JAK3 and / or TYK2 binder. In a particular embodiment, the Janus family kinase binder is a TYK2 binder. In a particular embodiment, the Janus family kinase binder is a dual binder of TYK2 and JAK1.

[0106] Examples of Tyk2 selective binders / inhibitors include deucravacitinib (BMS- 986165) (MedChemExpress, HY-117287), BMS-986202 (MedChemExpress, HY-131968), lomedeucitinib (BMS-986322) (MedChemExpress, HY-160144S), ropsacitinib (Selleckchem, S9676), GLPG3667 (MedChemExpress, HY-156961), zasocitinib (TAK-297)(MedChemExpress, HY-150096) and ABBV-712 (MedChemExpress, HY-156409) and derivatives thereof.

[0107] Examples of Tyk2 Jak1 dual binders include SAR-20347 (MedChemExpress, HY-100895), brepocitinib (PF-06700841) (MedChemExpress, HY-112708), and derivatives thereof.

[0108] In some embodiments, the TM is a compound of formula (IIA) or (IIB):(HA) (IIB) wherein R1and R2are each independently H, Ci-4alkyl, X is CH or N;Y is CH or N;Rbis Ci-ealkylene optionally interrupted with one or more heteroatoms selected from O and NR6where R6is H, or Ci-4alkyl;and5represents the point of attachment to L1.

[0109] In some embodiments, the TYK2 / JAK1 binder is a compound of formula (IIC):(IIC) wherein R1and R2are each independently H, Ci-4alkyl, and represents the point of attachment to L1.

[0110] In some embodiments, R1is H, methyl or ethyl.

[0111] The compound of formula (ii) can be a Tyk2 binder or a dual Tyk2 Jak1 binder.

[0112] In some embodiments, the E3L comprises a piperazine group, optionally a phenyl piperazine group or derivatives thereof. In some embodiments, E3L is a cereblon E3 ligase ligand, for example, thalidomide, lenalidomide, pomalidomide or a derivative with an N- alkylated glutarimide ring.

[0113] In some embodiments, the E3L a E3 ligase ligand for VHL, DCAF11, DDB1 or CUL4. Examples include ligands described by Liu et al. (PMID: 37845222).

[0114] In some embodiments, the linker is a peptidic linker comprising at least one amino acid. In some embodiments, the linker L1is an amino acid that results in an amide linkage between TM and E3L. Without wishing to be bound to theory, such a linker may more readily break up upon catabolism in the liver by microsomal amidases and various CYPs / UGT, which may reduce any potential for liver-associated toxicity.

[0115] In some embodiments, the linker is a self-immolative linker.

[0116] The linker, particularly L1, can comprise amino acids L-Lysine, L-Ornithine (L-Orn), and / or L-diaminobutyric acid (L-Dab), or D-amino acid analogues of the above.

[0117] In some embodiments, L1is prepared using a compound selected from the group consisting of Fmoc-Lys(Boc)-OPfp, Fmoc-Orn(Boc)-Opfp, and Fmoc-Dab(Boc)-Opfp. For example, Fmoc-Phe-Lys(Boc)-PAB-PNP is a cleavable linker compound and can be used in making the compounds described herein.

[0118] In some embodiments, wherein L1is represented by Formula (III A):wherein R3is absent, NR6or NH;R4is absent, Ci-ealkylene optionally interrupted with one or more heteroatoms selected from O and NR6, piperazinyl, piperidinyl, phenylpiperazinyl or phenylpiperid inyl;R5is Ci-salkylene, optionally interrupted with one or more heteroatoms selected from O andNR6;R6is H or Ci-4alkyl; and represent points of attachment to the remainder of the compound of Formula (I) where * is the attachment to TM, ** is the attachment to E3L and *** is the attachment to L2.

[0119] In some embodiments, L1is represented by Formula (IIIB):wherein R3is absent, NR6or NH; R7is CH or N;R5is Ci-salkylene, optionally interrupted with one or more heteroatoms selected from O and NR6;R6is H or Ci-4alkyl; and represent points of attachment to the remainder of the compound of Formula (I) where * is the attachment to ITM, ** is the attachment to E3L and *** is the attachment to L2.

[0120] In some embodiments, L1is represented by Formula (IIIC):wherein R3is absent, NR6or NH;R4is absent, Ci-ealkylene optionally interrupted with one or more heteroatoms selected from O and NR6, piperazinyl, piperidinyl, phenylpiperazinyl or phenylpiperidinyl;R5is Ci-salkylene, optionally interrupted with one or more heteroatoms selected from O and NR6;R6and R8are independently H or Chalky I; and represent points of attachment to the remainder of the compound of Formula (I) where * is the attachment to TM, ** is the attachment to E3L and *** is the attachment to L2.

[0121] In some embodiments, L2is stable to cleavage following subcutaneous injection. Such linkers can be stable to cleavage by proteases and stable to cleavage in the lymphatics system. Proteases and elastases can be released in the skin following and as aresult of subcutaneous injection. In some embodiments, L2is a disulfide-based linker, including hindered disulfide linkers. Disulfide based linkers can be reduced for example by intracellular or extracellular glutathione. In other embodiments, such linkers include non-cleavable linkers, such as PEG linkers. For example, PEG- and disulfide-based linkers are not cleavable by proteases or elastases. PEG-based linkers can release their payload following for example lysosomal degradation of the linked antibody.

[0122] In some embodiments, L2is a peptidic (also referred to as peptide-based) linker. Peptidic or peptide based linkers for example can undergo proteolysis in the lysosome releasing their payload.

[0123] In some embodiments, L2is selected from the group consisting of SPDP,SPDB, Sulfo-SPDP, SPDB-sulfo, Sulfo-SPDB, DMAC-SPDB, SMAC-SPDB-sulfo, SPP, DMAC-SPP, Sulfo-SPP, SPDMV, SPDMV-sulfo, Py-ds-dmBut-OPfp, Mal-PEG1-NHS, PySS- PEG2-NHS, Mal-Ala-Ala-PAB-PNP, MP-Ala-Ala-PAB-PNP, MC-Ala-Ala-PAB-PNP, Mal-Val- Ala-PAB-PNP, MP-Val-Ala-PAB-PNP, and MC-Val-Ala-PAB-PNP. In some embodiments, L2is selected fromwherein n is an integer from 1 to 10.

[0124] In some embodiments, L2is selected fromwherein Rxand Ry are independently selected from L-Ala, D-Ala, Gly, D-Glu or L-Glu. The foregoing are examples of self-immolating linkers.

[0125] In some embodiments, L2comprises a reactive moiety for reacting with a binding molecule, which can be selected from maleimide derivatives, disulfide derivatives, 10 pyridylthiol derivatives, thiol derivatives, vinyl sulfone derivatives, aziridine derivatives or haloacetamide derivatives.

[0126] In some embodiments, the conjugation reagent is selected from:

[0127] In some embodiments, the conjugation reagent is selected from:a stereoisomer, a pharmaceutically acceptable salt, and / or solvate thereof.

[0128] In some embodiments, the conjugation reagent is selected from:

[0129] In some embodiments, the conjugation reagent is selected from compounds I- 5, I-6, I-7, I-8, I-9, 1-10, and 1-21.

[0130] In some embodiments, TM has a binding affinity of at least 10 nM, for example about 10 nM to about 500 nM.

[0131] In some embodiments, TM has a binding affinity of less than 1 nM, for example about 0 nM to about 1 nM.

[0132] In some embodiments, TM has a binding affinity of at least 1 nM, for example about 1 nM to about 10 nM.

[0133] In some embodiments, TM has a binding affinity of at least 100 nM, for example about 100 nM to about 200 nM.

[0134] In some embodiments, TM has a binding affinity of at least 200 nM, for example about 200 nM to about 300 nM.

[0135] In some embodiments, TM has a binding affinity of at least 300 nM, for example about 300 nM to about 400 nM.

[0136] In some embodiments, TM has a binding affinity of at least 400 nM, for example about 400 nM to about 500 nM or more than about 500 nM.

[0137] Also provided in another aspect is a targeted degrader conjugate (also referred to as a DAC) comprising, the conjugation reagent conjugated to a CD127 binding agent (BA), wherein the CD127 binding agent is covalently attached to L2.

[0138] In some embodiments, the conjugation reagent is conjugated to a CD127 binding agent, wherein the CD127 binding agent is covalently attached to L2by reacting with the reactive moiety of L2.

[0139] In some embodiments, the targeted degrader conjugate is represented by Formula (IV):TM - L1- E3LL I2B IA(IV) wherein TM is a Janus family kinase domain-targeting binder; L1is a linker; L2is a binding molecule linker; E3L is a E3 ligase ligand; and BA is a CD127 binding agent; wherein, TM, L2and E3L are covalently linked to L1and L2is covalently linked to the CD127 binding agent.

[0140] In some embodiments, the CD127 binding agent is an antibody, a peptide, or a peptide mimetic.

[0141] In an embodiment, the CD127 binding agent is an antibody or binding fragment thereof. In one embodiment, the antibody is a bispecific antibody that binds CD127 and a non- CD127 antigen.

[0142] In some embodiments, the binding agents such as antibodies, do not induce antibody-dependent cell-mediated cytotoxicity (ADCC) and / or complement-dependent cytotoxicity or complement activation. The antibodies described herein can be in some embodiments lgG4 isotype and can in some embodiments have a human lgG4 framework. The lgG4 antibody constant regions can be human native sequences (without stabilizing mutations to prevent half-body exchange). Alternatively, they can comprise one or more mutations. For example, the lgG4 half-body exchange can be prevented by a number of constant region mutations. For example, the S228P mutation of the lgG4 constant region can prevent half-body exchange. Other mutations of the lgG4 that may be useful include E233P, F234V, F234A, L235A, D265A, R409K, K196Q, F296Y, E356K, H435R, L445P, G446> del, and / or K447> del. An example lgG4 isotype backbone that can be used is that of the gemtuzumab antibody (which is useful as an ADC) and harbors the S228P mutation. The IgG 1 isotype with mutations that remove ADCC and complement activation activities (Fc-disabled or F-silenced) can also be used, for example, the N297A mutation. Other examples include one or more of the following mutations: L234F, L235E, L235Q, M252Y, S254T, T256E, K322Q, P331S.

[0143] The CD127 binding agent (BA) may permit for targeting of specific T cell subset or sub-populations, such as T-helper and / or T -follicular helper cells, Tern (T effector memory cells), Tern (T central memory cells). In some embodiments, TB binds some T cells but not others, for example, effector T cells but not regulatory T cells (Treg).

[0144] The CD127 binding agent can be an anti-CD127 specific antibody (i.e. CD127 antibody). The CD127 antibody can interfere with ligand binding (IL7 and / or TSLP binding) be specific for an epitope that comprises one or more ligand binding loops (L1-L6), specifically, (defined in PMID: 22308406) in the mature protein (with signal sequence cleaved), L1 (aa31- 34), L2 (aa56-61), L3 (aa77-84), L4 (aa101-106), L5 (aa137-141), L6 (aa190-193). The CD127 antibody can be specific for an epitope outside the ligand binding site (of IL7 and / or TSLP ligands) selectively on the outside of or opposite side to the known ligand binding site formedby residues in the D1 and D2 domains of the extracellular portion of CD127. Optionally, an epitope that does not include all or any amino acids from the L1 through L6 elbow loops that can be involved in ligand binding. Optionally, an epitope that excludes one or more of the following amino acids: Ser51, His53, Ala76, Leu77, Val78, Glu79, Val80, Lys97, Lys98, Phe99, Leu100, Leu101 , lle102, Gly103, Asp122, Thr124, Arg140, Gly142, Ala143, Asp145, Lys158, Tyr159, Hist 80, Val181 , Asn182, Leu183, Ser184, Ser185, Thr186, Lys187, Leu188, Thr189, Leu190, Leu191 , Arg193, His211, Tyr212, Phe213. Optionally, an epitope that includes one or more of the following amino acids: Ser3, Phe4, Thr81 , Lys83, Lys84, Met128, Ile172, Pro173, Asp174, Hist 75, Phe177, Lys178, Trp181 , Ser225, Tyr227, Leu237, Tyr240, Gly241, Ser282, Val289, Val290, Ser334, Tyr335, Ala336, Trp350, Tyr362, Try363, Ala364, Asp365, Lys368, Lys401, Phe408, Asp409. The CD127 antibody can also be specific for an epitope outside the homodimer or heterodimer interface of CD127 formed with one or more interacting receptor chains (CD132, TSLPR). For example, the CD127 can be a non-agonistic and non-antagonistic internalizing anti-CD127 antibody. For example, antibodies that are not antagonistic or agonistic have been described in the literature. Anti-CD127 antibodies have been described that bind outside and on the opposite side of the D1 and D2 domains of the extracellular region of the IL-7RA chain that bind the IL-7 ligand, and which are capable of efficiently internalizing into cells (see for example, PMID: 30850736). The anti-CD127 antibody can also be an antagonistic internalizing antibody, targeting CD127, optionally targeting the CD127 extracellular membrane-proximal domain (approx, residues 130-230). In an embodiment, the anti-CD127 antibody does not antagonize or agonize the thymic stromal lymphopoietin receptor (TSLPR), of which the IL-7RA chain is also a co-receptor.

[0145] In an embodiment, the calculated hydrophilicity / hydrophobicity of the conjugation reagent or the payload fragment (expressed by ClogP) is less than 4, preferably less than 2, or preferably less than 1. ClogP can be determined for example using algorithms such as the one provided by BioByte.

[0146] In another embodiment, the hydrophilicity / hydrophobicity of the payload fragment released from the targeted degrader conjugate when processed inside the cell (wherein the BA (e.g. targeting antibody) is cleaved off and / or degraded) is less than 1, preferably less than 0. Payload fragments released in a cell can be cysteine or lysine complexed or complexed with another amino acid. For example, the payload fragment derived from targeted degrader conjugate comprising compound 1-17 may be released from the antibody, via proteolytic degradation of the antibody component, as a cysteine-complexed payload or compound of the following formula with ClogP of -1 .72:

[0147] The hydrophilicity of the conjugated payload resulting from the conjugation reagent can be estimated by ClogP (calculated logP value) as developed by BioByte. For compounds 1-1 through I-20 (non-cysteine complexed), the following table estimates the relative ClogP values:

[0148] For payloads resulting from compounds 1-21 to I-29, additionally to ClogP, tPSA and logS, the following table also depicts PAMPA permeability estimates at pH7.4 (physiological) and pH5.0 (endosomal / lysosomal), with moderate to high values of PAMPA at pH5.0 relative to pH7.4 indicating compounds that are expected to more readily escape the endosomal compartment, an important attribute for greater activity for an antibody- delivered payload. Additionally, a relative ranking of the estimated cytosolic stability to human liver microsomal stability is shown. Values include calculations by ADME@NCATS tools.III. Antibodies and degrader antibody conjugates

[0149] Various anti-CD127 antibodies are described.

[0150] In some embodiments, the CD127 specific binding agent is an anti-CD127 antibody (e.g. antibody that specifically binds CD127).

[0151] In some embodiments, the antibody is an a CD127 / IL-7Ra specific antibody, optionally an agonistic antibody or an antagonist antibody. Anti-CD127 antibodies are known in the art. For example, anti-CD127 antibodies described in WO2022220625A1 , titled Protein degrader conjugates and use thereof, can be used.

[0152] They may also be prepared. For example, a CD127 specific antibody may be generated by immunizing a transgenic animal with CD127 extracellular domain or a portion thereof, optionally wherein the portion is amino acid residues 130-230 of UniProt ID: P16871. In a particular embodiment, the antibody specifically binds the CD127 extracellular membrane- proximal domain (e.g., residues 130-230 of the amino acid sequence UniProt ID: P16871 ). Methods of engineering antibodies are known to the person skilled in the art, and include for example, commercial antibody engineering, phage display methods, and immunization of transgenic mice.

[0153] Various procedures known in the art can be used for the production of antibodies against the target epitope of interest. Such antibodies include, but are not limited to, monoclonal antibodies, chimeric antibodies, single chain antibodies, Fab fragments, and fragments produced by Fab expression libraries.

[0154] For the production of antibodies, various host animals (including but not limited to rabbits, mice, rats, chicken, llama, camelid, etc.) are immunized by injection with the target protein of interest. Various adjuvants (depending on host species, Freund (complete and incomplete), mineral gels such as aluminum hydroxide, surfactants such as lysolecithin, pluronic polyols, polyanions, peptides, suspensions Immune response using oil, keyhole limpet hemocyanin, dinitrophenol, and potentially useful human adjuvants such as, but not limited to, BCG (bacille Calmette-Guerin) and Corynebacterium parvum).

[0155] Monoclonal antibodies against the target of interest can be prepared using any technique provided for the production of antibody molecules by continuous cell lines in culture.This technique includes the hybridoma technology first described by Kohler and Milstein (1975, Nature 256: 495-497), the human B cell hybridoma technology (Kosbor et al., 1983, Immunology Today 4:72; Cote et al., 1983, Proc. Natl. Acad. Sci. USA 80: 2026-2030) and EBV hybridoma technology (Cole et al., 1985, Monoclonal Antibodies and Cancer Therapy, Alan R. Liss, Inc. 77-96). It is not limited. Furthermore, a technology developed for the production of “chimeric antibodies” by splicing genes from appropriate antigen-specific mouse antibody molecules together with genes from appropriate biologically active human antibody molecules (Morrison et al., 1984, Proc. Natl. Acad. Sci. USA 81 : 6851-6855; Neuberger et al., 1984, Nature 312: 604-608; Takeda et al., 1985, Nature 314: 452-454). Alternatively, techniques described for the production of single chain antibodies (US Pat. No. 4,946,778) can be applied to the production of single chain antibodies with the desired specificity.

[0156] Antibody fragments that contain specific binding sites for the target protein of interest can be generated by known techniques. For example, such fragments include F (ab') 2 fragments that can be generated by pepsin digestion of antibody molecules, and Fab fragments that can be generated by reduction of disulfide bridges of F (ab') 2 fragments. It is not limited to these. Alternatively, Fab expression libraries can be constructed (Huse et al., 1989, Science 246: 1275-1281 ), allowing rapid and easy identification of monoclonal Fab fragments with the desired specificity for the target protein of interest.

[0157] In some embodiments, the antibodies described herein can be humanized. The humanization of antibodies from non-human species (for example from mouse or rabbit) has been well described in the literature. See for example EP-B1 0239400 and Carter P & Merchant AM, 1997, herein incorporated by reference in their entirety. Humanized antibodies are also readily obtained commercially (e.g., Scotgen Limited, 2 Holly Road, Twickenham, Middlesex, Great Britain).

[0158] Humanized forms of rodent antibodies are readily generated by CDR grafting (Riechmann L et al, 1998). In this approach the six CDR loops comprising the antigen binding site of the rodent monoclonal antibody are linked to corresponding human framework regions. CDR grafting often yields antibodies with reduced affinity as the amino acids of the framework regions may influence antigen recognition (Foote J & Winter G, 1992). To maintain the affinity of the antibody, it is often necessary to replace certain framework residues by site directed mutagenesis or other recombinant techniques and may be aided by computer modeling of the antigen binding site (Co MS et al., 1994).

[0159] Humanized forms of antibodies are optionally obtained by resurfacing (Pedersen JT et al., 1994). In this approach only the surface residues of a rodent antibody are humanized.

[0160] In some embodiments, the CD127-specific antibody is selected from antibodies designated 9B7, 6C5, 6A3, R34.34, GR34 and 1A11 or a binding fragment of any thereof, and associated humanized or chimeric thereof, analogs thereof, and antigen-binding fragments thereof, as described in W02010017468A1 , and the associated humanized antibodies described in WO2011094259A2. Sections related to the aforementioned antibodies and binding fragments thereof, including sequences, are incorporated herein by reference. For example,Humanized 1A11 clone CDRs:CDRH1 : GYTMN (SEQ ID NO: 1 )CDRH2: LINPYNGVTSYNQKFK (SEQ ID NO: 2)CDRH3: GDGNYWYFDV (SEQ ID NO: 3)CDRL1:SASSSVTYMH (SEQ ID NO: 4)CDRL2: EISKLAS (SEQ ID NO: 5)CDRL3: QEWNYPYT (SEQ ID NO: 6) using the Kabat numbering system.

[0161] In an embodiment, the CD127-specific antibody is a humanized antibody comprising for example, CDRs (SEQ ID NO: 1-6) grafted onto a human lgG4 / kappa framework, optionally in a nivolumab framework. In some embodiments, the CD127 specific antibody has the heavy chain variable domain shown in SEQ ID NO: 8, the heavy chain shown in SEQ ID NO: 7, the light chain variable domain shown in SEQ ID NO: 10 or the light chain shown in SEQ ID NO: 9.QVQLVESGGGVVQPGRSLRLDCKASGITFSGYTMNWVRQAPGKGLEWVALINPYNGVTSY NQKFKGRFTISRDNSKNTLFLQMNSLRAEDTAVYYCATGDGNYWYFDVWGQGTLVTVSSAS TKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYS LSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPK PKDTLMISRTPEVTC VVVD VSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNS TYR VVS VLT VLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLV KGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHE ALHNHYTQKSLSLSLGK (SEQ ID NO: 7)QVQLVQSGAEVKKPGASVKVSCKASGYTFTGYTMNWVRQAPGQGLEWMGLINPYNGVTS YNQKFKGRVTLTRDTSISTAYMELSRLRSDDTAVYYCARGDGNYWYFDVWGQGTTVTVSS (SEQ ID NO: 8)EIVLTQSPATLSLSPGERATLSCSASSSVTYMHWYQQKPGQAPRLLIYEISKLASGIPARFSGS GSGTDFTL.TISSLEPEDFAVYYCQEWNYPYTFGQGTKVEIKR T AAPS 'F / FPPSDEQZ-KSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHK VYACEVTHQGLSSPVTKSFNRGEC (SEQ ID NO: 9)EIVLTQSPATLSLSPGERATLSCSASSSVTYMHWYQQKPGQAPRPLIYEISKLASGIPARFSG SGSGTDFTLTISSLEPEDFAVYYCQEWNYPYTFGGGTKVEIK ( SEQ ID NO: 10)

[0162] In some embodiments, the CD127 specific antibody is a variant of clone 1 A11, optionally BPC4398 (1A11.H3L4 HCDR3.N98D, i.e. HCDR3 is GDGDYWYFDV (SEQ ID NO: 11 ). Said variant is reported to have an improved affinity overthe parental molecule 1 A11 H3L4.

[0163] In some embodiments, the CD127 specific antibody is a single chain antibody.In some embodiments, the single chain antibody is a scFV.

[0164] In some embodiments, the CD127 specific antibody has the CDRs:CDRH1 SYAMS ( SEQ ID NO 12)CDRH2 AISGSGGSTYYADSVKG ( SEQ ID NO 13)CDRH3 WVSLPTFDY ( SEQ ID NO 14)CDRL1 QGDSLRSYYAS ( SEQ ID NO 15)CDRL2 GKNNRPS ( SEQ ID NO 16)CDRL3 NSSDVHMPYW ( SEQ ID NO 17) using the Kabat numbering system.

[0165] In some embodiments, the CD127 specific antibody comprises CDRs of SEQ ID Nos: 12-17 in a human lgG4 / kappa framework.

[0166] In some embodiments, the CD127 specific antibody has heavy chain shown in SEQ ID NO: 18, and the light chain shown in SEQ ID NO: 19 or 20.QVQLVESGGGVVQPGRSLRLDCKASGITFSSYAMSWVRQAPGKGLEWVAAISGSGGSTYY ADSVKGRFTISRDNSKNTLFLQMNSLRAEDTAVYYCATWVSLPTFDYWGQGTLVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGL YSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFP PKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSV LTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTC LVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVM HEALHNHYTQKSLSLSLGK (SEQ ID NO: 18)SSELTQDPAVSVALGQTVRITCQGDSLRSYYASWYQQKPGQAPVLVIYGKNNRPSGIPDRFSGSSSGNTASLTITGAQAEDEADYYCNSSDVHMPYVVFGGGTKLTVLGQPKANPTVTLFPPSSEELQANKATLVCLISDFYPGAVTVAWKADGSPVKAGVETTKPSKQSNNKYAASSYLSLTPEQWKSHRSYSCQVTHEGSTVEKTVAPTECS (SEQ ID NO: 19)QSALTQPASVSGSPGQSITISCQGDSLRSYYASWYQQHPGKAPKLMIYGKNNRPSGVSNRFSGSKSGNTASLTISGLQAEDEADYYCNSSDVHMPYVVFGTGTKVTVLGQPKA / VPTVTLFPPSSEELQANKATLVCLISDFYPGAVTVAWKADGSPVKAGVETTKPSKQSNNKYAASSYLSLTPE QWKSHRSYSCQVTHEGSTVEKTVAPTECS (SEQ ID NO: 20)

[0167] In some other embodiments, the CD127 specific antibody has the CDRs:CDRH1 DYYMH (SEQ ID NO 21 )CDRH2 YIYPSNGGNGYNQKVKG (SEQ ID NO 22)CDRH3 GTYYDGSYFDV (SEQ ID NO 23)CDRL1 KASQDVSTTLA (SEQ ID NO 24)CDRL2 SASYRYT (SEQ ID NO 25)CDRL3 QQHYSIPRT (SEQ ID NO 26) using the Kabat numbering system.

[0168] In some embodiments, the CD127 specific antibody comprises CDRs 21-26 in a human lgG4 / kappa framework.

[0169] In some embodiments, the CD127 specific antibody has heavy chain shown in SEQ ID NO: 27 or its variable domain, and the light chain shown in SEQ ID NO: 28 or its variable domain.QVQLVESGGGVVQPGRSLRLDCKASGITFSDYYMHWVRQAPGKGLEWVAYIYPSNGGNGY NQKVKGRFTISRDNSKNTLFLQMNSLRAEDTAVYYCATGTYYDGSYFDVWGQGTLVTVSS ASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGL YSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSV LTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTC LVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVM HEALHNHYTQKSLSLSLGK fSEQ ID NO: 27)EIVLTQSPATLSLSPGERATLSCKASQDVSTTLAWYQQKPGQAPRLLIYSASYRYTGIPARFS GSGSGTDFTLTISSLEPEDFAVYYCQQHYSIPRTFGQGTKVEIKR7VAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDS TYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC fSEQ ID NO: 28)

[0170] In the foregoing sequences and sequences described herein, the variable domain is shown without italics and the italicized portion is the remainder of the chain. CDR sequences may be underlined and bolded.

[0171] For example, the CD127 specific antibody is an antibody that targets activated T cells, effector memory T cells (Tern) and central memory T cells (Tern) and does not bind regulatory ? cells (Tregs).

[0172] In some other embodiments, the CD127 specific antibody has the CDRs:CDR-H1: GYTMN (SEQ ID NO: 29)CDR-H2: LINPYNGVTSYNQKFKG (SEQ ID NO: 30)CDR-H3: GDGNYWYFDV (SEQ ID NO: 31)CDR-L1 : SASSSVTYMH (SEQ ID NO: 32)CDR-L2: EISKLAS (SEQ ID NO: 33) andCDR-L3: QEWNYPYT (SEQ ID NO: 34) using the Kabat numbering system.

[0173] In some embodiments, the CD127 specific antibody comprises CDRs 29-34 in a human lgG4 / kappa framework.

[0174] In some embodiments, the CD127 specific antibody (GNP) has heavy chain variable shown in SEQ ID NO: 43, and the light chain variable domain shown in SEQ ID NO: 44. In some embodiments, the light chain variable domain can in addition to SEQ ID NO: 44 can include RTV.

[0175] In some embodiments, the CD127 specific antibody (GNP) has heavy chain shown in SEQ ID NO: 75, and the light chain shown in SEQ ID NO: 76.QVQLVQSGAEVKKPGASVKVSCKASGYTFTGYTMNWVRQAPGQGLEWMGLINPYNGVTS YNQKFKGRVTLTRDTSISTAYMELSRLRSDDTAVYYCARGDGNYWYFDVWGQGTTVTVSSA STKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLY SLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVL TVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCL VKGFYPSDIA VEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCS VMH EALHNHYTQKSLSLSLGK (SEQ ID NO: 75)EIVLTQSPATLSLSPGERATLSCSASSSVTYMHWYQQKPGQAPRPLIYEISKLASGIPARFSG SGSGTDFTLTISSLEPEDFAVYYCQEWNYPYTFGGGTKVEIKRT AAPSV,F / FPPSDEQ / _KSG TASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKH KVYACEVTHQGLSSPVTKSFNRGEC (SEQ ID NO: 76)

[0176] In some other embodiments, the CD127 specific antibody (GDS) has the CDRs:CDR-H1: GYTMN (SEQ ID NO: 37)CDR-H2: LINPYNGVTSYNQKFKG (SEQ ID NO: 38)CDR-H3: GDGDYWYFDV (SEQ ID NO: 39)CDR-L1 : SASSSVTYMH (SEQ ID NO: 40)CDR-L2: EISKLAS (SEQ ID NO: 41) andCDR-L3: QEWNYPYT (SEQ ID NO: 42) using the Kabat numbering system.

[0177] In some embodiments, the CD127 specific antibody comprises CDRs 37-42 in a human lgG4 / kappa framework.

[0178] In some embodiments, the CD127 specific antibody (GDS) has heavy chain variable domain shown in SEQ ID NO: 51 , and the light chain variable domain shown in SEQ ID NO: 52. In some embodiments, the light chain variable domain can in addition to SEQ ID NO: 52 include RTV. In some embodiments, the CD127 specific antibody (GDS) has heavy chain shown in SEQ ID NO: 77, and the light chain shown in SEQ ID NO: 78.QVQLVQSGAEVKKPGASVKVSCKASGYTFTGYTMNWVRQAPGQGLEWMGLINPYNGVTS YNQKFKGRVTLTRDTSISTAYMELSRLRSDDTAVYYCARGDGDYWYFDVWGQGTTVTVSSA STKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLY SLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPSCPAPEFLGGPSVFLFPP KPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVL TVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCL VKGFYPSDIA VEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCS VMH EALHNHYTQKSLSLSLGK (SEQ ID NO: 77)EIVLTQSPATLSLSPGERATLSCSASSSVTYMHWYQQKPGQAPRPLIYEISKLASGIPARFSG SGSGTDFTLTISSLEPEDFAVYYOQEWNYPYTFGGGTKVEIKRT AAPSV,F / FPPSDEQ / _ / <SG TASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC (SEQ ID NO: 78)

[0179] In some other embodiments, the CD127 specific antibody (PHP) has theCDRs:CDR-H1: SYAMS (SEQ ID NO: 45)CDR-H2: AISGSGGSTYYADSVKG (SEQ ID NO: 46)CDR-H3: WVSLPTFDY (SEQ ID NO: 47)CDR-L1 : QGDSLRSYYAS (SEQ ID NO: 48)CDR-L2: GKNNRPS (SEQ ID NO: 49) andCDR-L3: NSSDVHMPYVV (SEQ ID NO: 50) using the Kabat numbering system.

[0180] In some embodiments, the CD127 specific antibody comprises CDRs 45-50 in a human lgG4 / lambda framework.

[0181] In some embodiments, the CD127 specific antibody (PHP) has heavy chain variable domain shown in SEQ ID NO: 59, and the light chain variable domain shown in SEQ ID NO: 6O._ln some embodiments, the light chain variable domain can in addition to SEQ ID NO: 60 include GQP.

[0182] In some embodiments, the CD127 specific antibody (PHP) has heavy chain shown in SEQ ID NO: 79, and the light chain shown in SEQ ID NO: 80.EVQLLESGGGLVQPGGSLRLSCAASGFTFSSYAMSWVRQAPGKGLEWVSAISGSGGSTYY ADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAKWVSLPTFDYWGQGTLVTVSSAST KG PS VFPLAPCSRSTSES TAALGCL VKD YFPEPVTVSWNSGALTSGVHTFPA VLQSSGL YSL SSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKP KDTLMISRTPEVTC VVVD VSQEDPEVQFNWYVDG VEVHNAKTKPREEQFNS TYR VVS VL TV LHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVK GFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGK (SEQ ID NO: 79)SSELTQDPAVSVALGQTVRITCQGDSLRSYYASWYQQKPGQAPVLVIYGKNNRPSGIPDRFSGSSSGNTASLTITGAQAEDEADYYCNSSDVHMPYVVFGGGTKLTVLGQPKA / VPTVTLFPPSSEELQANKA TL VCLISDFYPGA VTVA WKADGSP VKA G VETTKPSKQSNNKYAASS YLSL TPEQWKSHRSYSCQVTHEGSTVEKTVAPTECS (SEQ ID NO: 80)

[0183] In some other embodiments, the CD127 specific antibody (PHS) has theCDRs:CDR-H1: SYAMS (SEQ ID NO: 53)CDR-H2: AISGSGGSTYYADSVKG (SEQ ID NO: 54)CDR-H3: WVSLPTFDY (SEQ ID NO: 55)CDR-L1 : QGDSLRSYYAS (SEQ ID NO: 56)CDR-L2: GKNNRPS (SEQ ID NO: 57) andCDR-L3: NSSDVHMPYVV (SEQ ID NO: 58) using the Kabat numbering system.

[0184] In some embodiments, the CD127 specific antibody comprises CDRs 53-58 in a human lgG4 / lambda framework.

[0185] In some embodiments, the CD127 specific antibody (PHS) has heavy chain variable domain shown in SEQ ID NO: 67, and the light chain variable domain shown in SEQ ID NO: 68. In some embodiments, the light chain variable domain can in addition to SEQ ID NO: 68 include GQP.

[0186] In some embodiments, the CD127 specific antibody (PHS) has heavy chain shown in SEQ ID NO: 81, and the light chain shown in SEQ ID NO: 82.EVQLLESGGGLVQPGGSLRLSCAASGFTFSSYAMSWVRQAPGKGLEWVSAISGSGGSTYY ADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAKWVSLPTFDYWGQGTLVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGL YSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPSCPAPEFLGGPSVFLFP PKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSV LTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVM HEALHNHYTQKSLSLSLGK (SEQ ID NO: 81)SSELTQDPAVSVALGQTVRITCQGDSLRSYYASWYQQKPGQAPVLVIYGKNNRPSGIPDRFSGSSSGNTASLTITGAQAEDEADYYCNSSDVHMPYVVFGGGTKLTVLGQPKA / VPT TLFPPSSEELQANKA TL VCLISDFYPGA VTVA WKADGSP VKA G VETTKPSKQSNNKYAASS YLSL TP EQWKSHRSYSCQVTHEGSTVEKTVAPTECS (SEQ ID NO: 82)

[0187] In some other embodiments, the CD127 specific antibody (MOP) has theCDRs:CDR-H1: DYYMH (SEQ ID NO: 61 )CDR-H2: YIYPDNGGNGYNQKFKG (SEQ ID NO: 62)CDR-H3: GTYYDGSYFDY (SEQ ID NO: 63)CDR-L1 : KASQDVSTTVA (SEQ ID NO: 64)CDR-L2: SASYRYT (SEQ ID NO: 65) andCDR-L3: QQHYSIPRT (SEQ ID NO: 66) using the Kabat numbering system.

[0188] In some embodiments, the CD127 specific antibody comprises CDRs 61-66 in a human lgG4 / kappa framework.

[0189] In some embodiments, the CD127 specific antibody (MCP) has heavy chain variable domain shown in SEQ ID NO: 75, and the light chain variable domain shown in SEQ ID NO: 76. In some embodiments, the light chain variable domain can in addition to SEQ ID NO: 76 include RTV.

[0190] In some embodiments, the CD127 specific antibody (MOP) has heavy chain shown in SEQ ID NO: 83, and the light chain shown in SEQ ID NO: 84.EVQLQQSGPELVKPGASVKMSCKASGYTFSDYYMHWVKQSHGKSLEWIGYIYPDNGGNGYNQKFKGKATLTVDKSSSTVYMELRSLTSEDSALYYCARGTYYDGSYFDYWGQGTTLTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLY SLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVL TVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCL VKGFYPSDIA VEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCS VMHEALHNHYTQKSLSLSLGK (SEQ ID NO: 83)DIVMTQSHKFMSTLVGDRVSITCKASQDVSTTVAWYQQKPGQSPKLLIYSASYRYTGVPDRFTGSGSGTDFTFTISSVQAEDLAVYYCQQHYSIPRTFGGGTKLEIKRT AAPSV'F / FPPSDEQ / .KSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYE KHKVYACEVTHQGLSSPVTKSFNRGEC (SEQ ID NO: 84)

[0191] In some other embodiments, the CD127 specific antibody (MHP) has the CDRsCDR-H1: SYAMS (SEQ ID NO: 69)CDR-H2: AISGSGGSTYYADSVKG (SEQ ID NO: 70)CDR-H3: WVSLPTFDY (SEQ ID NO: 71 )CDR-L1 : KASQDVSTTLA (SEQ ID NO: 72)CDR-L2: SASYRYT (SEQ ID NO: 73); andCDR-L3: QQHYSIPRT (SEQ ID NO: 74) using the Kabat numbering system.

[0192] In some embodiments, the CD127 specific antibody comprises CDRs 69-74 in a human lgG4 / kappa framework.

[0193] In some embodiments, the CD127 specific antibody (MHP) has heavy chain variable domain shown in SEQ ID NO: 35, and the light chain variable domain shown in SEQ ID NO: 36. In some embodiments, the light chain variable domain can in addition to SEQ ID NO: 36 include RTV.

[0194] In some embodiments, the CD127 specific antibody has heavy chain shown inSEQ ID NO: 85, and the light chain shown in SEQ ID NO: 86.QVQLVQSGAEVKKPGASVKVSCKASGYSFTDYYMHWVRQAPGQRLEWMGYIYPSNGGNG YNQKVKGRVTITRDTSASTAYMELSSLRSEDTAVYYCVRGTYYDGSYFDVWGQGTLVTVSS ASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFP PKPKDTLMISRTPEVTCVVVD VSQEDPEVQFNWYVDG VEVHNAKTKPREEQFNS TYR VVS V LTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTC LVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVM HEALHNHYTQKSLSLSLGK (SEQ ID NO: 85)DIQMTQSPSSLSASVGDRVTITCKASQDVSTTLAWYQQKPGKAPKLLIYSASYRYTGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQHYSIPRTFGPGTKVEIKRT AAPSV'F / FPPSDEQ / .KSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC (SEQ ID NO: 86)

[0195] In some embodiments, the sequences can comprise S228P (shown in italics, in red and underline in the heavy chain sequences herein or serine at this position.

[0196] In an embodiment, the antibody comprises a heavy chain variable region comprising: i) an amino acid sequence as set forth in SEQ ID NO: 35, ii) an amino acid sequence with at least 80%, at least 90%, or at least 95% sequence identity to SEQ ID NO: 35, wherein the CDR sequences are as set forth in SEQ ID NOs: 29-31 , and / or wherein the antibody comprises a light chain variable region comprising an amino acid sequence as setforth in SEQ ID NO: 36, ii) an amino acid sequence with at least 80%, at least 90%, or at least 95% sequence identity to SEQ ID NO:36, wherein the CDR sequences are as set forth in SEQ ID NOs: 32-34.

[0197] In an embodiment, the antibody comprises a heavy chain variable region comprising: i) an amino acid sequence as set forth in SEQ ID NO: 43, ii) an amino acid sequence with at least 80%, at least 90%, or at least 95% sequence identity to SEQ ID NO: 43, wherein the CDR sequences are as set forth in SEQ ID NOs:37-39, and / or wherein the antibody comprises a light chain variable region comprising an amino acid sequence as set forth in SEQ ID NO: 44, ii) an amino acid sequence with at least 80%, at least 90%, or at least 95% sequence identity to SEQ ID NO:44, wherein the CDR sequences are as set forth in SEQ ID NOs: 40-42.

[0198] In an embodiment, the antibody comprises a heavy chain variable region comprising: i) an amino acid sequence as set forth in SEQ ID NO: 51 , ii) an amino acid sequence with at least 80%, at least 90%, or at least 95% sequence identity to SEQ ID NO: 51, wherein the CDR sequences are as set forth in SEQ ID NOs: 45-47, and / or wherein the antibody comprises a light chain variable region comprising an amino acid sequence as set forth in SEQ ID NO: 52, ii) an amino acid sequence with at least 80%, at least 90%, or at least 95% sequence identity to SEQ ID NO: 52, wherein the CDR sequences are as set forth in SEQ ID NOs: 48-50.

[0199] In an embodiment, the antibody comprises a heavy chain variable region comprising: i) an amino acid sequence as set forth in SEQ ID NO: 59, ii) an amino acid sequence with at least 80%, at least 90%, or at least 95% sequence identity to SEQ ID NO: 59, wherein the CDR sequences are as set forth in SEQ ID NOs:53-55, and / or wherein the antibody comprises a light chain variable region comprising an amino acid sequence as set forth in SEQ ID NO: 60, ii) an amino acid sequence with at least 80%, at least 90%, or at least 95% sequence identity to SEQ ID NO:60, wherein the CDR sequences are as set forth in SEQ ID NOs: 56-58.

[0200] In an embodiment, the antibody comprises a heavy chain variable region comprising: i) an amino acid sequence as set forth in SEQ ID NO: 67, ii) an amino acid sequence with at least 80%, at least 90%, or at least 95% sequence identity to SEQ ID NO: 67, wherein the CDR sequences are as set forth in SEQ ID NOs:61-63, and / orwherein the antibody comprises a light chain variable region comprising an amino acid sequence as set forth in SEQ ID NO: 68, ii) an amino acid sequence with at least 80%, at least 90%, or at least 95% sequence identity to SEQ ID NO:68, wherein the CDR sequences are as set forth in SEQ ID NOs: 64-66.

[0201] In an embodiment, the antibody comprises a heavy chain variable region comprising: i) an amino acid sequence as set forth in SEQ ID NO: 75, ii) an amino acid sequence with at least 80%, at least 90%, or at least 95% sequence identity to SEQ ID NO: 75,wherein the CDR sequences are as set forth in SEQ ID NOs: 69-71 , and / or wherein the antibody comprises a light chain variable region comprising an amino acid sequence as set forth in SEQ ID NO: 76, ii) an amino acid sequence with at least 80%, at least 90%, or at least 95% sequence identity to SEQ ID NO:76, wherein the CDR sequences are as set forth in SEQ ID NOs: 72-74.

[0202] Other CD127 specific antibodies that can be used include, for example: PF- 06342674 / RN168 (as disclosed in US8298535B2), clone A7R34 (PMID: 8415665), OSE-703 (Effi-3) I lusvertikimab, ADX-914 (BMS-986265 I bempikibart) sections incorporated herein by reference as they relate to antibodies the recited antibodies, including binding fragments and sequences thereof.

[0203] In some embodiments, the CD127 specific antibody is clone 2B8 or clone 4A10 or a binding fragment of either thereof, as described in US11111306B2 and Hixon JA, et al. New anti-IL-7Ra monoclonal antibodies show efficacy against T cell acute lymphoblastic leukemia in pre-clinical models. Leukemia. 2020 Jan;34(1 ):35-49. doi: 10.1038 / s41375-019- 0531-8. Epub 2019 Aug 22. PMID: 31439943; PMCID: PMC8132108, sections incorporated herein by reference as they relate to antibodies 2B8 and 4A10, including binding fragments and sequences thereof.

[0204] In some embodiments, the CD127 specific antibody is clone N13B2-G4 or a binding fragment thereof, as described in US11440964B2, hereby incorporated by reference as it relates to the aforementioned antibodies, including sequences thereof.

[0205] In some embodiments, the CD127 specific antibody is clone MD707-1 (deposited at the CNCM under NO 1-4531 ), MD707-3 (deposited at the CNCM under No. I- 4532) or MD707-13 (deposited at the CNCM under No. I-4533), or a binding fragment of any thereof, as described in WQ2013 / 056984, incorporated by reference as it relates to the aforementioned antibodies.

[0206] In some embodiments, the CD127 antibody is clone B12 (sequence as provided in Figure S1 ) or a binding fragment thereof as described in Akkapeddi, P., Fragoso, R., Hixon, J. A. et a / . A fully human anti-IL-7Ra antibody promotes antitumor activity against T- cell acute lymphoblastic leukemia. Leukemia 33, 2155-2168 (2019). https: / / doi.Org / 10.1038 / s41375-019-0434-8, hereby incorporated by reference as it relates to antibody B12 and its binding fragments, including its sequence.

[0207] The antibodies and other binding agents described herein can be used to measure the number of CD127+ T cells and / or soluble CD127 level in a sample.

[0208] Accordingly also provided are methods of identifying if a subject is afflicted with a CD127 mediated disorder, the method comprising measuring the level of CD127+, optionally CD127+CD44+ T cells and / or soluble CD127 using a binding agent, optionally an antibody described herein.

[0209] In some embodiments, the CD127-specific binding agent is an IL-7 mimetic, optionally a peptide mimetic.

[0210] In some embodiments, the CD127-specific binding agent is an IL-7 mimetic or IL-7 receptora (IL-7Ra) ligand. The IL-7Ra ligand may for example be a molecule that binds the IL-7Ra chain as disclosed in US11254729 For example, the IL-7Ra ligand can be the molecules identified as MDK-703, and MDK-1472. These molecules are engineered peptide mimetics that are unrelated in structure tolL-7. MDK-703 comprises MDK-1472 fused to an lgG2 Fc-domain, as described in Dower WJ, Park Al, Bakker AV, Cwirla SE, Pongtornpipat P, Williams BM, et al. (2023) A mechanistically novel peptide agonist of the IL-7 receptor that addresses limitations of IL-7 cytokine therapy, PLoS ONE 18(10): e0286834. https: / / doi.orci / 10.1371 / iournal.pone.0286834. incorporated herein by reference, insofar as they relate to the IL-7 mimetic, or IL-7Ra ligand sequence. The IL-7Ra ligand may comprise a sequence as described in Dower et al., and US11254729B2, incorporated herein by reference insofar as it relates to the IL-7Ra ligand.

[0211] Other binding proteins can also be used. For example, the binding protein may be an Anti-CD45RA antibody (a marker of CD4 TEMRA cells; and having low expression on subsets of active Treg cells; broad expression on other cells - monocytes and granulocytes).

[0212] In some embodiments, the binding protein is an anti-CCR7 antibody. CCR7 is expressed on most peripheral T cells and exhibits low expression on a subset of effector / memory-like Treg cells that infiltrate inflamed sites and tumors).

[0213] Also provided in another aspect are compositions comprising the conjugation reagent or targeted degrader conjugates described herein. The composition may comprise a pharmaceutically acceptable carrier.

[0214] Pharmaceutical compositions can include, without limitation, lyophilized powders or aqueous or non-aqueous sterile injectable solutions or suspensions, which may further contain antioxidants, buffers, bacteriostats and solutes that render the combinations or compositions substantially compatible with the tissues or the blood of an intended recipient. Other components that may be present in such combinations or compositions include water, surfactants (such as Tween™), alcohols, polyols, glycerin and vegetable oils, for example. Extemporaneous injection solutions and suspensions may be prepared from sterile powders, granules, tablets, or concentrated solutions or suspensions. The combination or compositionmay be supplied, for example but not by way of limitation, as a lyophilized powder which is reconstituted with sterile water or saline or other pharmaceutically acceptable diluent prior to administration to the patient.

[0215] Pharmaceutical compositions may comprise a pharmaceutically acceptable carrier. Suitable pharmaceutically acceptable carriers include essentially chemically inert and nontoxic compositions that do not interfere with the effectiveness of the biological activity of the pharmaceutical composition. Examples of suitable pharmaceutical carriers include, but are not limited to, water, saline solutions, glycerol solutions, ethanol, N-(1(2,3-dioleyloxy)propyl)N,N,N- trimethylammonium chloride (DOTMA), diolesylphosphotidyl-ethanolamine (DOPE), and liposomes. Such combinations or compositions should contain a therapeutically effective amount of the compound, together with a suitable amount of carrier so as to provide the form for direct administration to the patient.

[0216] In addition, there is provided a pharmaceutical formulation comprising the conjugation reagent or the targeted degrader conjugate, or a derivative, analog, pharmaceutically acceptable salt thereof or a mixture of any of the foregoing and optionally a pharmaceutically acceptable carrier. One or more molecules of the disclosure can be present in association with one or more non-toxic pharmaceutically acceptable carriers and / or diluents and if desired other active ingredients. The pharmaceutical compositions of the disclosure can be in a form suitable for parenteral or systemic use.

[0217] Suitable diluents for polypeptides, including antibodies and / or conjugates comprising antibodies include but are not limited to saline solutions, pH buffered solutions and glycerol solutions or other solutions suitable for freezing polypeptides and / or cells.

[0218] The combinations or compositions described herein can be prepared by known methods for the preparation of pharmaceutically acceptable compositions that can be administered to subjects such that an effective quantity of the active substance is combined in a mixture with a pharmaceutically acceptable vehicle.

[0219] The term “degrader payload” as used herein refers to a fragment of the targeted degrader conjugate where the BA has been cleaved or degraded.

[0220] In some embodiments, the targeted degrader conjugate’s metabolites upon cleavage of the binding agent are selected from:

[0221] In some embodiments, the targeted degrader conjugate’s metabolites upon cleavage of the binding agent are selected from:Methods of Making

[0222] Another aspect provides a method of making compounds described herein. In general, conjugation reagents may be prepared and isolated by synthetic methods known to those skilled in the art for analogous compounds and by methods described in detail in the Examples and Schemes herein. One skilled in the art would appreciate that, where a particular protecting group, leaving group, reactive group or transformation condition is used, other protecting groups, leaving groups, reactive groups and transformation conditions may also be suitable and contemplated.

[0223] In certain embodiments, compounds of the present invention are generally prepared synthetically by coupling their respective components, E3L, TM, L1and L2, as follows:

[0224] Above, E3L can be coupled with L1using any or a combination or a sequence of Schemes I, II and III, or a portion of Schemes XIII and XV. TM can be synthesized using any or a combination or a sequence of Schemes IV, V, IX, X, XI, XII, or a portion of Schemes XVIII. E3L-L1can be coupled with TM using any or a combination or a sequence of Schemes VI, or a portion of Schemes XVI or XVIII. E3L-U-TM (degrader conjugate reagent) can be coupled via L1to L2using Scheme VII, or a portion of Schemes XVI, XVIII, XIX, XX, or XXI. E3L-L1(L2)-(TM) can be coupled to Antibody via L2using Scheme VIII. The above compounds can also be prepared by following Schemes XIII, XV, XIV, XVI, XVII, XVIII, XIX, XX, and XXI.

[0225] Degrader conjugation reagent (E3L-L1(L2)-(TM) with a maleimide or pyridyl disulfide sulfhydryl reactive group as part of L2can be coupled to an antibody via intrachain disulfides using the following method: Antibody is first reduced with TCEP (for example, using 4:1 molar equivalents of TCEP to antibody in phosphate or another compatible buffer at neutral pH without divalent cations and incubation over 4 hours at 37°C) in the presence of EDTA or DTPA. The antibody is then purified by desalting. The intrachain disulfide-reduced antibody is then reacted with a degrader conjugation reagent in a 1-1 Ox molar excess in DMA at a final concentration of 10-20% DMA for 1-2 hours at room temperature. Maleimide-based conjugation reaction can be quenched with 10x molar excess of N-acetyl cysteine. Conjugate is purified using a desalting column, for example PD10 column with Sephadex™ G-25 resin, or ultrafiltration / diafiltration (UF / DF). Conjugation can be evaluated by measuring UV / Vis absorbance at 280nm and 330nm.

[0226] Degrader conjugation reagent (E3L-L1-TM) with a maleimide or pyridyl disulfide sulfhydryl reactive group can also be coupled to an antibody to random primary amines with the use of 2-iminothionate (Traut’s reagent). The antibody is first modified with 10x molar equivalents of freshly-made T raut’s reagent in a phosphate, borate or another compatible buffer at neutral pH and incubated for 1-2 hours at room temperature protected from light. The reaction can be quenched with lysine. The antibody is then cleaned up by desalting. The sulfhydryl modified antibody is then reacted with a degrader conjugation reagent in a 1 -1 Ox molar excess in DMA at a final concentration of 10-20% DMA for 1-2 hours at room temperature. Maleimide- based conjugation reaction can be quenched with 10x molar excess of N-acetyl cysteine. Conjugate is purified using a desalting column, for example PD10 column with Sephadex G-25 resin, or ultrafiltration / diafiltration (UF / DF). Conjugation can be evaluated by measuring UV / Vis absorbance at 280nm and 330nm.Methods of Use

[0227] Also provided are methods of using and uses of the degrader compounds described herein.

[0228] Accordingly, an aspect includes a method of treating an autoimmune disorder, the method comprising administering an effective amount of the targeted degrader construct described herein or a composition comprising the targeted degrader construct to a subject in need thereof.

[0229] Another aspect includes a method of treating a CD127 mediated disorder, the method comprising administering an effective amount of the targeted degrader construct described herein or a composition comprising the targeted degrader construct to a subject in need thereof.

[0230] In some embodiments, the autoimmune disorder or the CD127 mediated disorder is rheumatoid arthritis (RA), systemic lupus erythematosus (SLE), celiac disease (CD), inflammatory bowel disease (IBD), ulcerative colitis (UC), psoriasis (PsO), psoriatic arthritis (PsA), diabetes, including T1 D, T2D, immune checkpoint inhibitor-induced type 1 diabetes or multiple sclerosis (MS). In an embodiment, the autoimmune disorder or the CD127 mediated disorder is atopic dermatitis (AD). In some embodiments, the autoimmune disorder or the CD127 mediated disorder is rheumatoid arthritis (RA). In some embodiments, the autoimmune disorder or the CD127 mediated disorder is systemic lupus erythematosus (SLE). In some embodiments, the autoimmune disorder or the CD127 mediated disorder is celiac disease (CD). In some embodiments, the autoimmune disorder or the CD127 mediated disorder is inflammatory bowel disease (IBD). In some embodiments, the autoimmune disorder or the CD127 mediated disorder is ulcerative colitis (UC). In some embodiments, the autoimmune disorder or the CD127 mediated disorder is psoriasis (PsO). In some embodiments, the autoimmune disorder or the CD127 mediated disorder is psoriatic arthritis (PsA).

[0231] In some embodiments, the autoimmune disorder or the CD127 mediated disorder is atopic dermatitis (AD). In some embodiments, the autoimmune disorder or the CD127 mediated disorder is diabetes, including T1 D, T2D, immune checkpoint inhibitor- induced type 1 diabetes. In some embodiments, the autoimmune disorder or the CD127 mediated disorder is multiple sclerosis (MS).

[0232] In some embodiments, the subject is determined to have an increased number of CD127+ T cells, optionally CD127+CD44+ T cells and / or an increased level of soluble CD127. In some embodiments a sample from the subject eg. a blood sample is measured for numbers of CD127+ T cells, optionally CD127+CD44+ T cells and / or level of soluble CD127. Subject with an increased number of CD127+ T cells, optionally CD127+CD44+ T cells and / or an increased level of soluble CD127 can be treated with a targeted degrader construct, described herein. The sample is for example a blood sample, a plasma sample or a serum sample. The sample can for example be contacted with a CD127 binding agent (e.g. antibodydescribed herein) for example using flow cytometry, FACS, ELISA or other similar type method to measure the number of CD127+ T cells, optionally CD127+CD44+ T cells and / soluble CD127. Such subjects may experience greater benefits from treatment with the targeted degrader constructs described. The levels can also be used to monitor treatment. For example a sample can be taken after initiating treatment (or before and after initiating treatment) or at any time during treatment.

[0233] As demonstrated herein, the targeted degrader constructs are able to dramatically reduce fecal Iipocalin2 levels, TYK2 levels and JAK1 levels in a human IBD animal model. The human IBD animal model comprises donor CD4+ CD45RBh'9hT cells from C57BL / 6-I I7r™1<IL7R) ECD transgenic female mice transferred into a RAG1 KO recipients (on a C57BL / 6 background).

[0234] JAK1 and TYK2 signaling are important in diseases such as TA, SLE, CD, UC, PsO and PsA as well as diabetes. For example, TYK-2 inhibition delays type 1 diabetes onset in mice (DOI: 10.1016 / j.ebiom.2025.105734) and Jak inhibitor, baricitinib, are effective disease modifying treatments for several autoimmune diseases (N Engl J Med 2023;389:2140-2150 DOI: 10.1056 / NEJMoa2306691).

[0235] Follicular helper T cells (Tfh), which are CD127+ are also known to play a role in diabetes (Xuan Du, Yan Zhu, et al, Regulation of the Function of T Follicular Helper Cells and B Cells in Type 1 Diabetes Mellitus by the CX40 / CX40L Axis, The Journal of Clinical Endocrinology & Metabolism, Volume 109, November 2024, Pages 2823- 2830, https: / / doi.org / 10. 1210 / clinem / dqae248Y

[0236] Immune checkpoint inhibitor induces type 1 diabetes in1 -2% of checkpoint inhibitor treated patients. Tfh and JAK1 signaling have been implicated (JCI Insight. 2025; 10(13): e188843. https: / / doi.Org / 10.1172 / jci. insight.188843). JAK inhibitors also demonstrate efficacy in T2DM, potentially attributable to an autoimmune component of the inflammatory response (Front Endocrinol (Lausanne). 2019 Jul 4;10:451. doi: 10.3389 / fendo.2019.00451).

[0237] In some embodiments, the targeted degrader construct or the composition is formulated for parental administration, optionally intravenous, intramuscular, intraperitoneal or subcutaneous administration, optionally subcutaneous administration. In other embodiments, it is formulated for oral administration, for example when the CD127 binding agent is not an antibody.

[0238] In some embodiments, the targeted degrader construct or the composition is administered parentally. For example, in some embodiments, the administration is intravenous. In some embodiments the administration is sub-cutaneous.

[0239] The above disclosure generally describes the present application. A more complete understanding can be obtained by reference to the following specific examples. These examples are described solely for the purpose of illustration and are not intended to limit the scope of the application. Changes in form and substitution of equivalents are contemplated as circumstances might suggest or render expedient. Although specific terms have been employed herein, such terms are intended in a descriptive sense and not for purposes of limitation.ExamplesExample 1

[0240] Certain subsets of cells have been suggested to play an increased or particular role in the initiation or progression of various autoimmune disorders. The described drug degrader compounds are directed to targeting specific subsets of T cells as a more precise strategy for amelioration of these conditions, while minimizing treatment-induced side effects

[0241] CD127 (IL7RA) receptor chain (UniProtKB / Swiss-Prot ID P16871). CD127 is largely expressed on the T- and natural killer (NK)- cell compartments and is upregulated on activated T-helper (Th) cells and is downregulated on T-regulatory (Treg) cells. CD127 is known to internalize, which internalization could be increased by IL7 signaling, and is known to be degraded both in the lysosomes and by the 20S proteasomal machinery. Targeting the JAK1 / TYK2 signaling via the CD127 receptor may avoid the undesirable inhibition of these pathways in myeloid cells (DCs and macrophages) and Treg cells, as well as other cells that are implicated in the mechanisms of side-effects observed with untargeted JAK1 / TYK2 inhibitors.

[0242] The degrader payload is designed for specific release upon the internalization of the CD127-targeting antibody into the endosomal / lysosomal compartment, and is further designed to facilitate lysosomal escape (avoid endosomal entrapment), allowing the active payload to be released into the cytoplasm to facilitate the recruitment of the Cereblon E3 ligase (CRBN) to ubiquitinate JAK1 / TYK2 molecules and target them for degradation by the proteasomal machinery, in a self-catalyzing manner. The heterobifunctional degrader payload is designed to bind JAK1 / TYK2 JH2 domains with a target affinity range of 10-500nM with one end and CRBN with an affinity of greater than 100-200nM on the other end (with slow dissociation rates for CRBN preferred). Affinity can be measured using SPR or BLI methods.Exemplary synthetic schemesScheme I

[0243] tert-Butyl 4-(2-(2,6-dioxopiperidin-3-yl)-1 -oxoisoindolin-4-yl)piperazine-1 -carboxylate (1-1)

[0244] 3-(4-Bromo-1-oxoisoindolin-2-yl)piperidine-2, 6-dione was reacted with tertbutyl piperazine-1-carboxylate to form intermediate tert-butyl 4-(2-(2,6-dioxopiperidin-3-y l)-1 - oxoisoindolin-4-yl)piperazine-1-carboxylate (1-1 ).

[0245] 3-(1-Oxo-4-(piperazin-1-yl)isoindolin-2-yl)piperidine-2, 6-dione (1-2)

[0246] tert-Butyl 4-(2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)piperazine-1- carboxylate (1-1 ) is deprotected to form intermediate 3-(1-oxo-4-(piperazin-1-yl)isoindolin-2- yl)piperidine-2, 6-dione (1-2).Scheme II

[0247] (9H-Fluoren-9-yl)methyl tert-butyl (6-(4-(2-(2,6-dioxopiperidin-3-yl)-1- oxoisoindolin-4-yl)piperazin-1 -yl)-6-oxohexane-1 ,5-diyl)dicarbamate (1-3)

[0248] 3-(1-Oxo-4-(piperazin-1-yl)isoindolin-2-yl)piperidine-2, 6-dione (1-2) was reacted with perfluorophenyl N2-(((9H-fluoren-9-yl)methoxy)carbonyl)-N6-(tert-butoxycarbonyl)lysinate to form intermediate (9H-fluoren-9-yl)methyl tert-butyl (6-(4-(2-(2,6- dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)piperazin-1-yl)-6-oxohexane-1 ,5-diyl)dicarbamate (1-Scheme III

[0249] tert-Butyl (5-amino-6-(4-(2-(2,6-dioxopiperidin-3-yl)-1 -oxoisoindolin-4- yl)piperazin-1 -yl)-6-oxohexyl)carbamate (1 -4)

[0250] (9H-Fluoren-9-yl)methyl tert-butyl (6-(4-(2-(2,6-dioxopiperidin-3-yl)-1- oxoisoindolin-4-yl)piperazin-1-yl)-6-oxohexane-1,5-diyl)dicarbamate (1-3) is deprotected to form intermediate tert-butyl (5-amino-6-(4-(2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4- yl)piperazin-1-yl)-6-oxohexyl)carbamate (1-4).Bpin 1'51-6Scheme IV

[0251] 2-(2-Methoxy-3-nitrophenyl)-4,4,5,5-tetramethyl-1 ,3,2-dioxaborolane (1 -5)

[0252] 1-Bromo-2-methoxy-3-nitrobenzene was reacted with 4, 4, 4', 4', 5, 5, 5', 5'- octamethyl-2,2'-bi(1 ,3,2-dioxaborolane) to form intermediate 2-(2-methoxy-3-nitrophenyl)- 4,4,5,5-tetramethyl-1 ,3,2-dioxaborolane (1-5).

[0253] 3-(2-Methoxy-3-(4,4,5,5-tetramethyl-1 ,3,2-dioxaborolan-2-yl)phenyl)-1 - methyl-1H-1,2,4-triazole (1-6)

[0254] 2-(2-Methoxy-3-nitrophenyl)-4,4,5,5-tetramethyl-1 ,3,2-dioxaborolane (1-5) was reacted with 3-bromo-1-methyl-1 H-1 ,2,4-triazole to form intermediate 3-(2-methoxy-3- (4,4,5,5-tetramethyl-1 ,3,2-dioxaborolan-2-yl)phenyl)-1-methyl-1H-1,2,4-triazole (1-6).Scheme V

[0255] 6-Chloro-4-(2-methoxy-3-(1 -methyl-1 H-1 ,2,4-triazol-3-yl)phenyl)-N- methylnicotinamide (1-7)

[0256] 3-(2-Methoxy-3-(4,4,5,5-tetramethyl-1 ,3,2-dioxaborolan-2-yl)phenyl)-1- methyl-1H-1 ,2,4-triazole (1-6) was reacted with 4,6-dichloro-N-methylnicotinamide to form intermediate 6-chloro-4-(2-methoxy-3-(1-methyl-1 H-1 , 2,4-triazol-3-y I ) pheny l)-N- methylnicotinamide (1-7).

[0257] 6-((4-(2-Methoxy-3-(1 -methyl-1 H-1 ,2,4-triazol-3-yl)phenyl)-5-(methylcarbamoyl)pyridin-2-yl)amino)nicotinic acid (1-8)

[0258] 6-Chloro-4-(2-methoxy-3-(1-methyl-1 H-1 ,2,4-triazol-3-yl)phenyl)-N- methylnicotinamide (1-7) was reacted with 6-aminonicotinic acid to form intermediate 6-((4-(2- methoxy-3-(1-methyl-1H-1 ,2,4-triazol-3-yl)phenyl)-5-(methylcarbamoyl)pyridin-2- yl)amino)nicotinic acid (1-8).Scheme VI

[0259] tert-Butyl (6-(4-(2-(2,6-dioxopiperidin-3-yl)-1 -oxoisoindolin-4- yl)piperazin-1 -yl)-5-(6-((4-(2-methoxy-3-(1 -methyl-1 H-1 ,2,4-triazol-3-yl)phenyl)-5-(methylcarbamoyl)pyridin-2-yl)amino)nicotinamido)-6-oxohexyl)carbamate (1 -9)

[0260] 6-((4-(2-Methoxy-3-(1-methyl-1 H-1 ,2,4-triazol-3-yl)phenyl)-5-(methylcarbamoyl)pyridin-2-yl)amino)nicotinic acid (1-8) was reacted with tert-butyl (5-amino- 6-(4-(2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)piperazin-1-yl)-6-oxohexyl)carbamate (1- 4) to form intermediate tert-butyl (6-(4-(2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4- yl)piperazin-1-yl)-5-(6-((4-(2-methoxy-3-(1-methyl-1 H-1 ,2,4-triazol-3-yl)phenyl)-5- (methylcarbamoyl)pyridin-2-yl)amino)nicotinamido)-6-oxohexyl)carbamate (1-9).Scheme VII

[0261] 6-((5-((6-Amino-1-(4-(2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4- yl)piperazin-1-yl)-1-oxohexan-2-yl)carbamoyl)pyridin-2-yl)amino)-4-(2-methoxy-3-(1- methyl-1H-1,2,4-triazol-3-yl)phenyl)-N-methylnicotinamide (1 -10)

[0262] tert-Butyl (6-(4-(2-(2,6-dioxopiperidin-3-yl)-1 -oxoisoindolin-4-yl)piperazin-1 -yl)- 5-(6-((4-(2-methoxy-3-(1-methyl-1 H-1 ,2,4-triazol-3-yl)phenyl)-5-(methylcarbamoyl)pyridin-2- yl)amino)nicotinamido)-6-oxohexyl)carbamate (1-9) is deprotected to form intermediate 6-((5- ((6-amino-1-(4-(2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4-yl)piperazin-1-yl)-1-oxohexan-2-yl)carbamoyl)pyridin-2-yl)amino)-4-(2-methoxy-3-(1-methyl-1 H-1 ,2,4-triazol-3-yl)phenyl)-N- methylnicotinamide (1-10).

[0263] 4-((S)-2-((S)-2-(6-(2,5-Dioxo-2,5-dihydro-1 H-pyrrol-1 - yl)hexanamido)propanamido)propanamido)benzyl (6-(4-(2-(2,6-dioxopiperidin-3-yl)-1- oxoisoindolin-4-yl)piperazin-1 -yl)-5-(6-((4-(2-methoxy-3-(1 -methyl-1 H-1 ,2,4-triazol-3- yl)phenyl)-5-(methylcarbamoyl)pyridin-2-yl)amino)nicotinamido)-6-oxohexyl)carbamate (1-1)

[0264] 6-((5-((6-Amino-1-(4-(2-(2,6-dioxopiperidin-3-yl)-1-oxoisoindolin-4- yl)pi perazin-1 -yl)-1 -oxohexan-2-yl)carbamoyl)pyridin-2-yl)amino)-4-(2-methoxy-3-(1 -methyl- 1 H-1 ,2,4-triazol-3-yl)phenyl)-N-methylnicotinamide (1-10) was reacted with 4-((S)-2-((S)-2-(6- (2, 5-dioxo-2,5-dihydro-1 H-pyrrol-1 -yl)hexanamido)propanamido)propanamido)benzyl (4- nitrophenyl) carbonate (1-11 ) to form intermediate 4-((S)-2-((S)-2-(6-(2,5-dioxo-2,5-dihydro- 1 H-pyrrol-1 -yl)hexanamido)propanamido)propanamido)benzyl (6-(4-(2-(2,6-dioxopiperidin-3- yl)-1 -oxoisoindolin-4-yl)piperazin-1-yl)-5-(6-((4-(2-methoxy-3-(1 -methyl-1 H-1 , 2, 4-triazol-3- yl)phenyl)-5-(methylcarbamoyl)pyridin-2-yl)amino)nicotinamido)-6-oxohexyl)carbamate (1-1).Scheme VIII

[0265] 4-((2S)-2-((2S)-2-(6-(3-Disulfaneyl-2,5-dioxopyrrolidin-1- yl)hexanamido)propanamido)propanamido)benzyl (6-(4-(2-(2,6-dioxopiperidin-3-yl)-1- oxoisoindolin-4-yl)piperazin-1 -yl)-5-(6-((4-(2-methoxy-3-(1 -methyl-1 H-1 ,2,4-triazol-3- yl)phenyl)-5-(methylcarbamoyl)pyridin-2-yl)amino)nicotinamido)-6-oxohexyl)carbamate antibody conjugate (Abc-1)

[0266] 4-((S)-2-((S)-2-(6-(2,5-Dioxo-2,5-dihydro-1H-pyrrol-1- yl)hexanamido)propanamido)propanamido)benzyl (6-(4-(2-(2,6-dioxopiperidin-3-yl)-1- oxoisoindolin-4-yl)piperazin-1-yl)-5-(6-((4-(2-methoxy-3-(1-methyl-1H-1 ,2,4-triazol-3- yl)phenyl)-5-(methylcarbamoyl)pyridin-2-yl)amino)nicotinamido)-6-oxohexyl)carbamate (1-1) was reacted with an antibody to form 4-((2S)-2-((2S)-2-(6-(3-disulfaneyl-2,5-dioxopyrrolidin-1- yl)hexanamido)propanamido)propanamido)benzyl (6-(4-(2-(2,6-dioxopiperidin-3-yl)-1- oxoisoindolin-4-yl)piperazin-1-yl)-5-(6-((4-(2-methoxy-3-(1-methyl-1H-1 ,2,4-triazol-3- yl)phenyl)-5-(methylcarbamoyl)pyridin-2-yl)amino)nicotinamido)-6-oxohexyl)carbamate antibody conjugate (Abc-1 ).

[0267] The following are several alternative synthesis routes for Intermediate 2-7:Scheme IXScheme XScheme XII

[0268] The following specific compounds are given, synthesized from Intermediate 2-7 as a starting point:

[0269] Compound 1-7:Scheme XIII

[0270] Compound 1-6Scheme XV

[0271] Compound I-5:Scheme XIV

[0272] Compound 1-9Scheme XVI5

[0273] Alternate synthesis of compound 1-5, through synthesis of Intermediate 4-3:5 Scheme XVIII

[0274] Alternate synthesis of compound 1-6:Scheme XIX

[0275] Synthesis of compounds 1-7 and 1-8:Scheme XXScheme XXI

[0277] Synthesis of Compound 1-21 :Scheme XXII

[0278] Synthesis of Compound 1-28:Example 3

[0279] Experiments carried out to assess binding: In vitro binding of CD127 DAC candidates and corresponding monoclonal antibodies to several sub-types of human and mouse T cells (including Teff, Tern, Treg), and monocytes is assessed by flow cytometry. Mouse T cells can be derived from human ecdCD127 transgenic mice (available fromGemPharmatech and Biocytogen).

[0280] Experiments carried out to assess inhibition capacity of the CD127 DAC candidates: In vitro assays using IFNalpha (JAK1 , TYK2) and IFNgamma (JAK1, JAK2) stimulation are conducted for evaluating JAK / TYK2 inhibitors, with treatment with CD127 DAC candidates and corresponding monoclonal antibodies in human and mouse T cells (CD127+ve CD4 T cells and CD127-ive Treg), as well as control cells, monocytes and / or B cells. Analysis by Western blot for phosphorylated STATs (STAT1 / 1, STAT1 / 2), as well for JAK1 and TYK2 proteins is used to confirm degradation. Candidates are tested in vivo in SLE disease models. SLE disease models in humanized mice (NCG.huCD34) or in NZB / W or MRL-lpr strains of mice are treated with the DAC candidates. To induce SLE, either human SLE patient T cells areadoptively transferred or T cells from human ecdCD127 transgenic mouse (GemPharmatech or Biocytogen) are transferred, or chemically induced by IP injection of pristane (2,6,10, 14- tetramethylpentadecane / TMPD). Treatment comparatively evaluates CD127 DAC candidates, antibody controls, and small molecule inhibitor (JAKi) controls. SLE is induced in these models by the transfer of T cells. Spontaneous mouse models are also used, such as the BXSB mouse for mouse cross-reactive DACs or with the aid of an adoptive transfer from human ecdCD127 transgenic mice with a humanized ectodomain of CD127 (GemPharmatech or Biocytogen). Outcome measures of CD127 DACs include evaluating control of SLE manifestations and any unwanted side effects or toxicities.

[0281] Candidates are evaluated in vivo in IBD, UC and CD mouse models. In vivo treatment of IBD / UC / CD disease models is performed in humanized mice (NCG.huCD34) or in SCID or RAG(KO) strains of mice. To induce IBD, either human patient T cells are adoptively transferred or T cells from human ecdCD127 transgenic mouse (GemPharmatech or Biocytogen) are transferred, or chemically induced by IP injection of dinitrobenzene sulfonic acid (DNBS), dextran sulfate sodium (DSS), oxazolone, or trinitrobenzene sulfonic acid (TNBS). Treatment comparatively evaluates CD127 DAC candidates, antibody controls, and JAKi controls. IBD is induced in these models by the transfer of T cells. Outcome measures of CD127 DACs include evaluating improved control of IBD manifestations, while minimizing unwanted side effects or toxicities.

[0282] Candidates are evaluated in vivo in psoriasis (PsO) mouse models. Imiquimod (IMQ) cream is evenly applied to the shaved back skin of ecdCD127 mice (GemPharmatech or Biocytogen) once a day to induce psoriasis-like lesions. Treatment with CD12 DAC is administered and mice are monitored daily for body weight and clinical scores (erythema and scales and thickness). At the endpoint, mouse back skin is preserved in formalin to assess dorsal skin thickness and histology scores. Part of back skin is preserved to assess in situ proinflammatory cytokines. Other outcome measures at the endpoint include flow cytometry to test for T cell populations in the skin and blood, and their cytokine production.Example 4

[0283] Recombinant CD127 specific antibodies were made. The recombinant antibodies were expressed in a mammalian host cells (CHO cells). They were purified via AmMag™ Ultra AT Protein A MagBeads. The purity was measured at >80% purity via SDS- PAGE under non-reducing condition and >79% purity via SEC-HPLC. The amino acid sequences of the variable domain regions are shown in Table 1 with CDR1, CDR2 and CDR3 regions of heavy and light chains shown as underlined and bolded.Table 1 : Amino acid sequences of variable domain regionsExample 5

[0284] The antibodies described in Example 4 were used to measure CD127 expression in various in vitro cell models. Clones GNP, GDS, PHP, PHS, MCP, MHP, and a commercially available reagent antibody clone eBioRDR5 (Invitrogen, eBiosciences) were assessed. PMBCs, Molt-4 and Jurkat E6.1 cells were tested. The results are shown in Fig. 1 A to 1E. Fig. 1 A and 1 B show that a large proportion of CD3 positive PMBCs are CD127 positive. Fig. 1C to 1 E show that Molt-4 stained negative for CD127, with Jurkat E6.1 have moderate expression level. [Fig. 1 a shows cell models that are available with varying levels of expression, including low / negative.

[0285] The antibodies described in Example 4 were used to measure pHrodo dye- labelled anti-CD127 antibody internalization in Jurkat, Molt4 and PBMCs. The experiment set up is shown in Fig. 2. Black wall, clear bottom 96-well was coated with 0.1 mg / ml poly-D-lysine overnight at 4°C. The next day, the wells were washed with PBS 2 times, seeded with 0.1 M cells per well in Fluorobrite DMEM complete media, letting cells attach to the bottom for 3 hours in a 37°C incubator. Then 10ug / ml pHrodo dye labelled anti-CD127 antibody were added in Fluorobrite DMEM complete media and the plate was put in the Incucyte S3. Green and red fluorescence were monitored every 2 hours over 2 days. Each well has 4 scans to better capture the whole well.

[0286] The antibodies described in Example 4 were used to measure the internalization of Zenon-red (pHrodo™ iFL Red for human IgG labelling) labelled anti-CD127 antibodies using the set up in Fig. 2. The results are shown in Fig. 3A to 3F. Fig. 3A shows internalization of GNP 44 hours after adding 10ug / ml Zenon-red labelled anti-CD127 Ab to PMBCs, Fig. 3B shows internalization of GDS 44 hours after adding 10ug / ml Zenon-red labelled anti-CD127 Ab to PMBCs, Fig. 3C shows internalization of PHP 44 hours after adding 10ug / ml Zenon-red labelled anti-CD127 Ab to PMBCs, Fig. 3D shows internalization of PHS 44 hours after adding 10ug / ml Zenon-red labelled anti-CD127 Ab to PMBCs, Fig. 3E shows internalization of MCP 44 hours after adding 10ug / ml Zenon-red labelled anti-CD127 Ab to PMBCs, and Fig. 3F shows internalization of MHP 44 hours after adding 10ug / ml Zenon-red labelled anti-CD127 Ab to PMBCs.

[0287] The antibodies described in Example 4 were used to measure the internalization of Zenon-red labelled anti-CD127 antibodies using the set up in Fig. 2. PMBCswere tested. The results are shown in Fig. 4. Fig. 4 shows that GDS and PHP antibodies have better internalization than other antibodies tested, but all internalize over time.

[0288] The antibodies described in Example 4 were used to measure the internalization of pHrodo-green direct labelled anti-CD127 antibodies using the set up in Fig. 2. The results are shown in Fig. 5A to 5F. Fig. 5A shows internalization of GNP 44 hours after adding 10ug / ml pHrodo green labelled anti-CD127 Ab to PMBCs, Fig. 5B shows internalization of GDS 44 hours after adding 10ug / ml pHrodo green labelled anti-CD127 Ab to PMBCs, Fig. 5C shows internalization of PHP 44 hours after adding 10ug / ml pHrodo green labelled anti- CD127 Ab to PMBCs, Fig. 5D shows internalization of PHS 44 hours after adding 10ug / ml pHrodo green labelled anti-CD127 Ab to PMBCs, Fig. 5E shows internalization of MCP 44 hours after adding 10ug / ml pHrodo green labelled anti-CD127 Ab to PMBCs, and Fig. 5F shows internalization of MHP 44 hours after adding 10ug / ml pHrodo green labelled anti-CD127 Ab to PMBCs.

[0289] Antibodies made in Example 4 were used to prepare degrader constructs using conjugation compounds described in Example 2. Below table summarizes some examples of targeted degrader conjugates made:

[0290] The degrader constructs were used to measure the degradation of Jak1 and Tyk2 in Jurkat cells with small molecule compounds I-5, I-6, I-7, I-8, I-9 and 1-10 derived by quenching their sulfhydryl-reactive moieties. The results were shown in Fig. 6. Fig. 6 shows that Jak1 degradation was seen in I-5, I-6, I-8, I-9 and 1-10 degrader-treated Jurkat cells, andthat great Tyk2 degradation was seen in I-5, I-6, I-9 and 1-10 degrader-treated Jurkat cells, that pStatl signaling was also reduced by all 6 degraders, and that pStat3 signaling cannot be detected in resting state Jurkat cells.

[0291] The degrader antibody conjugates were used to measure the degradation of Jak1 , Tyk2 in PBMCs treated with PHP anti-CD127 Ab DACs (with drug-to-antibody ratio (DAR) of approximately 1 ). Results were shown in Fig. 7. Fig. 7 shows that degrader conjugates with payloads I-5, I-6, I-7, I-8, I-9 and 1-10 were effective in degrading Tyk2. Fig. 7 shows that degrader conjugates with payloads I-5, I-6, I-7, I-8, l-9 and 1-10 show some degradation of Jak1. Fig. 7 shows that degrader conjugates with payloads I-5, I-6, I-7, I-8, I-9 and 1-10 were effective in reducing pStatl and pStat3 signaling.Example 6

[0292] As mentioned in Example 5, the antibodies described in Example 4 were used to make CD127 DAC degrader constructs.

[0293] These constructs have desirable properties.

[0294] These CD127 antibodies may target Teff cells and Tern cells, while sparing regulatory T cells (T regs). In some embodiments, the DAC construct included a Cereblon binder described herein along with a linker. The linker can be stable to proteases and the environment of the lymphatic system such that it is suitable for subcutaneous administration. Also for example, the linkers described herein can be more susceptible to microsomal catabolism relative to its cytosolic stability, accounting for the liver-mediated absorption, distribution, metabolism and excretion (ADME) of the conjugate and enabling the liver to quickly catabolize the degrader payload when clearing the conjugate, minimizing potential for any liver toxicities. In some embodiments, the compounds have one or more properties similar or with 10% range of the properties described herein, e.g. PAMPA calculated for example as described. The degrader structures describe herein for example have an integrated chemical moiety or set of moieties that may permit increased endosomal escape. The dual JAK / TYK family binder used in some embodiments herein is potent at inhibiting the JAK / STAT pathway.Further, the molecules described herein may have been engineered for a desirable hydrophilicity. For example the molecules may have a ClogP value as claimed with biophysical properties that minimize tox / bystander activity.Example 7

[0295] The degrader described in Example 1 was used to model a degrader payload model of the induced “Kiss of Death” (ternary complex) between Cereblon E3 ligase and Tyk2.The model is shown in Fig. 8. Fig. 8 shows an antibody with a Tyk2 binding via the JH2 domain, a Cereblon binding, and a linker to the antibody.Example 8

[0296] Human CD127+ T cells sorted from human PBMCs were treated with various DACs (analyzed at 72 hours post-treatment), and expression and activation levels of JAK1 , TYK2, STAT 1 and STAT3 were assayed by Western blot.

[0297] Fig. 9 shows that DAC treatment reduced levels of TYK2 and JAK1 as well as reduced the activation (phosphorylation) of downstream STAT-3 and STAT-1.Example 9IBP model - early administration

[0298] Approx. 0.5x106Donor CD4+ CD45RBhiahT cells from C57BL / 6-ll7r™1<IL7R) ECD transgenic mice were transferred into RAG1 KO recipients to generate a model of human IBD in mice, which were monitored for changes in weight and fecal lipocalin-2. Animals were treated as indicated (IV administration twice weekly for approx. 7 weeks), and additional markers of inflammation and cells were analyzed on Day 70.

[0299] As shown in Fig. 10, DAC treatment dramatically decreased the % weight change seen with vehicle control in the human IBD model. Unlike vehicle control and similar to healthy controls, DAC treated animals showed weight gain over the test period.

[0300] Fecal levels of lipocalin-2 are shown in Fig. 11. DAC-treated animals, relative to vehicle treated controls, showed a reduction and a substantially lower levels of the fecal inflammatory marker, lipocail-2 (shown on a logarithmic scale).Example 10IBD model - late administration to animals with severe disease

[0301] Another experiment was conducted where the DAC was first administered at 5 weeks post-transfer for approx. 3 weeks. As previously, approx. 0.5x10® Donor CD4+ CD45RBh'9hT cells from C57BL / 6-I I7r™1(|L7R> ECD transgenic mice were transferred into RAG1 KO recipients to generate a model of human IBD in mice, which were monitored for changes in weight and fecal lipocalin-2. Animals were treated as indicated in Fig.12, and additional markers of inflammation and cells were analyzed on Day 70.

[0302] Fig. 12 shows that DAC treatment started after severe disease onset where vehicle treated mice show a precipitous decrease in weight, is able to stabilize the recipient’s weight.

[0303] Fig. 13 shows a similar experiment using upadacitinib (brand name Rinvoq) which is a JAK inhibitor administered orally 3x per week at 10 mg / kg (estimated HED of 30 mg QD) starting at week 5 for approx. 3 weeks, was somewhat slower and less effective and maintaining animal weight than DAC administration.

[0304] Fecal lipocalin levels which are shown in Fig. 14 demonstrate that DAC treatment started even 5 weeks after cell transfer, is able to prevent severe colitis.

[0305] Fig. 15 shows that DAC treatment and Upadacitinib both reduced lipocalin-2 levels post treatment and that DAC reduction may be better sustained.

[0306] Surprisingly at 70 days post transfer, DAC treated animals showed negligible levels of CD127+ cells (mainly dominated by CD44+ memory cells), whereas vehicle treated animals had greater than 50% of donor cells with the CD44+ memory phenotype. Representative flow cytometry analyses shown with the respective animals also losing 13% of their weight in vehicle treated control whereas DAC treated subject shown gained 12.5% (Fig. 16).

[0307] Importantly as shown in Fig. 17, both early and late DAC treatment greatly reduced the percentage of donor T cells at day 70 in the colon and spleen of treated animals. DAC efficacy in reducing the donor cell numbers in colon and spleen was much greater than that seen with Upadacitinib treatment.

[0308] Fig. 18 shows representative colon histological sections capturing elements of disease progression and inflammation in T cell transfer colitis animal model of IBD. Animals were treated as indicated, and sections of colon were obtained on Day 70 and processed for histology. H&E sections of representative colon sections are shown at 10x magnification. Vehicle treated control animals are showing significant signs of severe disease, including chronic inflammation, substantial cell infiltrates including neutrophil infiltration, loss of goblet cells, tissue damage / ulceration. In contrast, DAC treated animals show substantial recovery from inflammation and reduction in histopathological markers of disease on average similar to healthy subjects, and upadacitinib treated animals showing a degree of recovery and somewhat reduced inflammation as well.

[0309] While the applicant's teachings described herein are in conjunction with various embodiments for illustrative purposes, it is not intended that the applicant's teachings be limited to such embodiments as the embodiments described herein are intended to be examples. On the contrary, the applicant's teachings described and illustrated herein encompass various alternatives, modifications, and equivalents, without departing from the embodiments described herein, the general scope of which is defined in the appended claims.

Claims

CLAIMS:

1. A conjugation reagent according to the formula (I), or a stereoisomer, a pharmaceutically acceptable salt, and / or solvate thereof:TM - L1- E3LL I2(I) wherein TM is a JANUS family kinase domain-targeting binder; L1is a first linker; L2is a second linker; and E3L is a E3 ligase ligand; wherein, TM, L2and E3L are covalently linked to L1, and L2comprises a reactive moiety for reacting with a binding molecule.

2. The conjugation reagent of claim 1 , wherein TM is a JAK1 , JAK2, JAK2 and / or TYK2 binder, optionally a JH2 pseudokinase domain targeting binder or a catalytic domain binder, optionally a dual TYK2 / JAK1 JH2 pseudokinase domain-targeting binder or a JAK1 JH1 catalytic domain binder.

3. The conjugation reagent of claim 1 or 2, wherein the TM is a compound of formula (IIA) or (IIB):(HA) (IIB) wherein R1and R2are each independently H, Ci-4alkyl,X is CH or N;Y is CH or N;Rbis Ci-ealkylene optionally interrupted with one or more heteroatoms selected from O and NR6where R6is H, or Ci-4alkyl;A is selected fromand ’ represents the point of attachment to L1.

4. The conjugation reagent of claim 1 or 2, wherein the TM is a compound of formula (IIC):wherein R1and R2are each independently H, or Ci-4alkyl, and represents the point of attachment to L1.

5. The conjugation reagent of claim 3 or 4, wherein R1is H, methyl or ethyl.

6. The conjugation reagent of any one of claims 1 to 5, wherein L1comprises a peptidic linker comprising at least one amino acid.

7. The conjugation reagent of claim 6, wherein the peptidic linker comprises at least one L-Lysine, L-Ornithine (L-Orn), L-diaminobutyric acid (L-Dab), D-Lysine, D-Ornithine, or D- diaminobutyric acid.

8. The conjugation reagent of any one of claims 1 to 7, wherein L1is represented by Formula (IIIA):wherein R3is absent, NR6or NH;R4is absent, Ci-ealkylene optionally interrupted with one or more heteroatoms selected from O and NR6, piperazinyl, piperidinyl, phenylpiperazinyl or phenylpiperidinyl;R5is Ci-salkylene, optionally interrupted with one or more heteroatoms selected from O andNR6;R6is H or Ci-4alkyl; and represent points of attachment to the remainder of the compound of Formula (I) where * is the attachment to TM, ** is the attachment to E3L and *** is the attachment to L2.

9. The conjugation reagent of any one of claims 1 to 7, wherein L1is represented byFormula (I I IB)(IIIB) wherein R3is absent, NR6or NH;R7is CH or N;R5is Ci-salkylene, optionally interrupted with one or more heteroatoms selected from O and NR6;R6is H or Ci-4alkyl; and represent points of attachment to the remainder of the compound of Formula (I) where * is the attachment to TM, ** is the attachment to E3L and *** is the attachment to L2.

10. The conjugation reagent of any one of claims 1 to 7, wherein L1is represented by Formula (IIIC):wherein R3is absent, NR6or NH;R4is absent, Ci-ealkylene optionally interrupted with one or more heteroatoms selected from O and NR6, piperazinyl, piperidinyl, phenylpiperazinyl or phenylpiperidinyl;R5is Ci-salkylene, optionally interrupted with one or more heteroatoms selected from O and NR6;R6and R8are independently H or Chalky I; and represent points of attachment to the remainder of the compound of Formula (I) where * is the attachment to TM, ** is the attachment to E3L and *** is the attachment to L2.

11. The conjugation reagent of any one of claims 1 to 10, wherein L2is a non-cleavable linker, a disulfide-based linker, or a peptide-based linker, optionally a self-immolative linker,optionally comprising succinimidyl 3-(2-pyridyldithio)propionate (SPDP), N-succinimidyl 4-(2- pyridyldithio)butanoate (SPDB), Sulfo-SPDP, SPDB-sulfo, Sulfo-SPDB, dimethylacetamide- SPDB (DMAC-SPDB), DMAC-SPDB-sulfo, N-succinimidyl 4-(2-pyridyldithio)pentanoate (SPP), DMAC-SPP, Sulfo-SPP, 2,5-dioxopyrrolidin-1-yl 4-methyl-4-(pyridin-2- yldisulfaneyl)pentanoate (SPDMV), SPDMV-sulfo, perfluorophenyl 4-methyl-4-(pyridin-2- yldisulfanyl)pentanoate (Py-ds-dmBut-OPfp), 2,5-Dioxopyrrolidin-1-yl 3-(2-(2,5-dioxo-2,5- dihydro-1 H-pyrrol-1-yl)ethoxy)propanoate (Mal-PEG1-NHS), 2, 5-dioxopyrrolidin-1 -yl-3-(2-(2- (pyridin-2-yldisulfaneyl)ethoxy)ethoxy) propanoate (PySS-PEG2-NHS), maleimide-Ala-Ala- para-aminobenzyl alcohol-para-nitrophenyl (Mal-Ala-Ala-PAB-PNP), maleimidopropionyl-Ala- Ala-PAB-PNP (MP-Ala-Ala-PAB-PNP), maleimidocaproyl-Ala-Ala-PAB-PNP (MC-Ala-Ala- PAB-PNP), Mal-Val-Ala-PAB-PNP, MP-Val-Ala-PAB-PNP, or MC-Val-Ala-PAB-PNP.

12. The conjugation reagent of any one of claims 1 to 11 , wherein L2is selected from:’ represents the point of attachment to L1.

13. The conjugation reagent of any one of claims 1 to 11 , wherein L2is selected from:wherein Rxand Ry are independently selected from L-Ala, D-Ala, Gly, D-Glu or L-Glu, and represents the point of attachment to L1.5 14. The conjugation reagent of any one of claims 1 to 13, wherein E3L iswherein5represents the point of attachment to L1.

15. The conjugation reagent of claim 1 , selected from:a stereoisomer, a pharmaceutically acceptable salt, and / or solvate thereof.

16. The conjugation reagent of claim 1 , selected from:

17. The conjugation reagent of any one of claims 1 to 16, wherein TM has a binding affinity of at least 10 nM, optionally about 10 nM to 500 nM.

18. The conjugation reagent of any one of claims 1 to 17, wherein E3L has a binding affinity 5 of at least 100 nM, optionally about 100 nM to 200 nM, or more than 200 nM.

19. A targeted degrader conjugate or a stereoisomer, a pharmaceutically acceptable salt, and / or solvate thereof, the conjugate comprising the conjugation reagent of any one of claims 1 to 18, conjugated to BA, wherein BA is a CD127 binding agent, wherein the CD127 binding agent is covalently attached to L2by reacting with the reactive moiety. 0 20. The targeted degrader conjugate of claim 19, having the Formula (IV):or a stereoisomer, a pharmaceutically acceptable salt, and / or solvate thereof.

21. The targeted degrader conjugate of claim 19 or 20, wherein the CD127 binding agent is an antibody, a peptide, or a peptide mimetic.

22. The targeted degrader conjugate of any one of claims 19 to 21 , wherein the CD127 binding agent is an antibody, or a binding fragment thereof.

23. The targeted degrader conjugate of claim 22, wherein the CD127-specific binding agent is a CD127 agonist or a CD127 antagonist.

24. The targeted degrader conjugate of claim 22, wherein the CD127-specific binding agent is an IL-7 peptide mimetic.

25. The targeted degrader conjugate of claim 22, wherein the CD127-specific binding agent is an antibody that comprises a heavy chain variable region and a light chain variable region, the heavy chain variable region comprising complimentary determining regions CDR- H1, CDR-H2 and CDR-H3, the light chain variable region comprising complementarity determining regions CDR-L1 , CDR-L2 and CDR-L3 and with the amino acid sequences of said CDRs comprising the sequences:CDR-H1: GYTMN (SEQ ID NO: 29)CDR-H2: LINPYNGVTSYNQKFKG (SEQ ID NO: 30)CDR-H3: GDGNYWYFDV (SEQ ID NO: 31)CDR-L1 : SASSSVTYMH (SEQ ID NO: 32)CDR-L2: EISKLAS (SEQ ID NO: 33); andCDR-L3: QEWNYPYT (SEQ ID NO: 34)CDR-H1: GYTMN (SEQ ID NO: 37)CDR-H2: LINPYNGVTSYNQKFKG (SEQ ID NO: 38)CDR-H3: GDGDYWYFDV (SEQ ID NO: 39)CDR-L1 : SASSSVTYMH (SEQ ID NO: 40)CDR-L2: EISKLAS (SEQ ID NO: 41)CDR-L3: QEWNYPYT (SEQ ID NO: 42)CDR-H1: SYAMS (SEQ ID NO: 45)CDR-H2: AISGSGGSTYYADSVKG (SEQ ID NO: 46)CDR-H3: WVSLPTFDY (SEQ ID NO: 47)CDR-L1 : QGDSLRSYYAS (SEQ ID NO: 48)CDR-L2: GKNNRPS (SEQ ID NO: 49)CDR-L3: NSSDVHMPYVV (SEQ ID NO: 50)CDR-H1: SYAMS (SEQ ID NO: 53)CDR-H2: AISGSGGSTYYADSVKG (SEQ ID NO: 54)CDR-H3: WVSLPTFDY (SEQ ID NO: 55)CDR-L1 : QGDSLRSYYAS (SEQ ID NO: 56)CDR-L2: GKNNRPS (SEQ ID NO: 57) andCDR-L3: NSSDVHMPYVV (SEQ ID NO: 58);CDR-H1: DYYMH (SEQ ID NO: 61 )CDR-H2: YIYPDNGGNGYNQKFKG (SEQ ID NO: 62)CDR-H3: GTYYDGSYFDY (SEQ ID NO: 63)CDR-L1 : KASQDVSTTVA (SEQ ID NO: 64)CDR-L2: SASYRYT (SEQ ID NO: 65) andCDR-L3: QQHYSIPRT (SEQ ID NO: 66); orCDR-H1: SYAMS (SEQ ID NO: 69)CDR-H2: AISGSGGSTYYADSVKG (SEQ ID NO: 70)CDR-H3: WVSLPTFDY (SEQ ID NO: 71)CDR-L1 : KASQDVSTTLA (SEQ ID NO: 72)CDR-L2: SASYRYT (SEQ ID NO: 73) andCDR-L3: QQHYSIPRT (SEQ ID NO: 74).

26. The targeted degrader conjugate of claim 25, wherein the antibody comprises a heavy chain variable region comprising: i) an amino acid sequence as set forth in SEQ ID NO: 35, ii) an amino acid sequence with at least 80%, at least 90%, or at least 95% sequence identity to SEQ ID NO: 35, wherein the CDR sequences are as set forth in SEQ ID NOs: 29-31, and / or wherein the antibody comprises a light chain variable region comprising an amino acid sequence as set forth in SEQ ID NO: 36, ii) an amino acid sequence with at least 80%, at least 90%, or at least 95% sequence identity to SEQ ID NO:36, wherein the CDR sequences are as set forth in SEQ ID NOs: 32-34; the antibody comprises a heavy chain variable region comprising: i) an amino acid sequence as set forth in SEQ ID NO: 43, ii) an amino acid sequence with at least 80%, at least 90%, or at least 95% sequence identity to SEQ ID NO: 43, wherein the CDR sequences are as set forth in SEQ ID NOs:37-39, and / or wherein the antibody comprises a light chain variable region comprising an amino acid sequence as set forth in SEQ ID NO: 44, ii) an amino acid sequence with at least 80%, at least 90%, or at least 95% sequence identity to SEQ ID NO:44, wherein the CDR sequences are as set forth in SEQ ID NOs: 40-42; the antibody comprises a heavy chain variable region comprising: i) an amino acid sequence as set forth in SEQ ID NO: 51, ii) an amino acid sequence with at least 80%, at least 90%, or at least 95% sequence identity to SEQ ID NO: 51 , wherein the CDR sequences are as set forth in SEQ ID NOs: 45-47, and / or wherein the antibody comprises a light chain variable region comprising an amino acid sequence as set forth in SEQ ID NO: 52, ii) an amino acid sequence with at least 80%, at least 90%, or at least 95% sequence identity to SEQ ID NO: 52, wherein the CDR sequences are as set forth in SEQ ID NOs: 48-50; the antibody comprises a heavy chain variable region comprising: i) an amino acid sequence as set forth in SEQ ID NO: 59, ii) an amino acid sequence with at least 80%, at least 90%, or at least 95% sequence identity to SEQ ID NO: 59, wherein the CDR sequences are as set forth in SEQ ID NOs:53-55, and / or wherein the antibody comprises a light chain variable region comprising an amino acid sequence as set forth in SEQ ID NO: 60, ii) an amino acid sequence with at least 80%, at least 90%, or at least 95% sequence identity to SEQ ID NO:60, wherein the CDR sequences are as set forth in SEQ ID NOs: 56-58;the antibody comprises a heavy chain variable region comprising: i) an amino acid sequence as set forth in SEQ ID NO: 67, ii) an amino acid sequence with at least 80%, at least 90%, or at least 95% sequence identity to SEQ ID NO: 67, wherein the CDR sequences are as set forth in SEQ ID NOs:61-63, and / or wherein the antibody comprises a light chain variable region comprising an amino acid sequence as set forth in SEQ ID NO: 68, ii) an amino acid sequence with at least 80%, at least 90%, or at least 95% sequence identity to SEQ ID NO:68, wherein the CDR sequences are as set forth in SEQ ID NOs: 64-66; or the antibody comprises a heavy chain variable region comprising: i) an amino acid sequence as set forth in SEQ ID NO: 75, ii) an amino acid sequence with at least 80%, at least 90%, or at least 95% sequence identity to SEQ ID NO: 75, wherein the CDR sequences are as set forth in SEQ ID NOs: 69-71 , and / or wherein the antibody comprises a light chain variable region comprising an amino acid sequence as set forth in SEQ ID NO: 76, ii) an amino acid sequence with at least 80%, at least 90%, or at least 95% sequence identity to SEQ ID NO:76, wherein the CDR sequences are as set forth in SEQ ID NOs: 72-74; optionally wherein the heavy chain and the light chain have the sequences of: SEQ ID NO: 75 and 76, 77 and 78, 79 and 80, 81 and 82, 83 and 84 or 85 and 86.

27. A composition comprising an effective amount of the conjugation reagent of any one of claims 1 to 18 or a stereoisomer, a pharmaceutically acceptable salt, and / or solvate thereof, or the targeted degrader conjugate of any one of claims 19 to 26 or a stereoisomer, a pharmaceutically acceptable salt, and / or solvate thereof, and a pharmaceutically acceptable carrier and / or diluent.

28. A method of treating an autoimmune disorder, the method comprising administering an effective amount of the targeted degrader construct of any one of claims 19 to 26 or a composition comprising the targeted degrader construct to a subject in need thereof.

29. The method of claim 28, wherein the autoimmune disorder is rheumatoid arthritis (RA), systemic lupus erythematosus (SLE), celiac disease (CD), inflammatory bowel disease (I BD), atopic dermatitis (AD) ulcerative colitis (UC), psoriasis (PsO), psoriatic arthritis (PsA), multiple sclerosis (MS), or diabetes, including T1 D, T2D, immune checkpoint inhibitor-induced type 1 diabetes.

30. The method of claim 28 or 29, wherein the composition is formulated for parental administration, optionally intravenous, intramuscular, intraperitoneal or subcutaneous administration, optionally subcutaneous administration.

31. An antibody comprising a light chain variable region and a heavy chain variable region, the heavy chain variable region comprising complimentary determining regions CDR- H1, CDR-H2 and CDR-H3, the light chain variable region comprising complementarity determining regions CDR-L1, CDR-L2 and CDR-L3 and with the amino acid sequences of said CDRs comprising the sequences:CDR-H1: SYAMS (SEQ ID NO: 69)CDR-H2: AISGSGGSTYYADSVKG (SEQ ID NO: 70)CDR-H3: WVSLPTFDY (SEQ ID NO: 71)CDR-L1 : KASQDVSTTLA (SEQ ID NO: 72)CDR-L2: SASYRYT (SEQ ID NO: 73) andCDR-L3: QQHYSIPRT (SEQ ID NO: 74).

32. An antibody comprising a light chain variable region and a heavy chain variable region, the heavy chain variable region comprising complimentary determining regions CDR- H1, CDR-H2 and CDR-H3, the light chain variable region comprising complementarity determining regions CDR-L1, CDR-L2 and CDR-L3, wherein the antibody comprises a heavy chain variable region comprising: i) an amino acid sequence as set forth in SEQ ID NO: 35, ii) an amino acid sequence with at least 80%, at least 90%, or at least 95% sequence identity to SEQ ID NO: 35, wherein the CDR sequences are as set forth in SEQ ID NOs: 29-31 , and / or wherein the antibody comprises a light chain variable region comprising an amino acid sequence as set forth in SEQ ID NO: 36, ii) an amino acid sequence with at least 80%, at least 90%, or at least 95% sequence identity to SEQ ID NO:36, wherein the CDR sequences are as set forth in SEQ ID NOs: 32-34.

33. An antibody comprising a light chain variable region and a heavy chain variable region, the heavy chain variable region comprising complimentary determining regions CDR-H1 , CDR- H2 and CDR-H3, the light chain variable region comprising complementarity determining regions CDR-L1, CDR-L2 and CDR-L3, wherein the antibody comprises a heavy chain variable region comprising: i) an amino acid sequence as set forth in SEQ ID NO: 43, ii) an amino acid sequence with at least 80%, at least 90%, or at least 95% sequence identity to SEQ ID NO: 43, wherein the CDR sequences are as set forth in SEQ ID NOs:37-39, and / or wherein the antibody comprises a light chain variable region comprising an amino acid sequence as set forth in SEQ ID NO: 44, ii) an amino acid sequence with at least 80%, at least 90%, or at least 95% sequence identity to SEQ ID NO:44, wherein the CDR sequences are as set forth in SEQ ID NOs: 40-42.

34. An antibody comprising a light chain variable region and a heavy chain variable region, the heavy chain variable region comprising complimentary determining regions CDR-H1 , CDR- H2 and CDR-H3, the light chain variable region comprising complementarity determining regions CDR-L1, CDR-L2 and CDR-L3, wherein the antibody comprises a heavy chain variable region comprising: i) an amino acid sequence as set forth in SEQ ID NO: 51, ii) an amino acid sequence with at least 80%, at least 90%, or at least 95% sequence identity to SEQ ID NO: 51, wherein the CDR sequences are as set forth in SEQ ID NOs:45-47, and / or wherein the antibody comprises a light chain variable region comprising an amino acid sequence as set forth in SEQ ID NO: 52, ii) an amino acid sequence with at least 80%, at least 90%, or at least 95% sequence identity to SEQ ID NO: 52, wherein the CDR sequences are as set forth in SEQ ID NOs: 48- 50.

35. An antibody comprising a light chain variable region and a heavy chain variable region, the heavy chain variable region comprising complimentary determining regions CDR-H1 , CDR- H2 and CDR-H3, the light chain variable region comprising complementarity determining regions CDR-L1 , CDR-L2 and CDR-L3, wherein, the antibody comprises a heavy chain variable region comprising: i) an amino acid sequence as set forth in SEQ ID NO: 59, ii) an amino acid sequence with at least 80%, at least 90%, or at least 95% sequence identity to SEQ ID NO: 59, wherein the CDR sequences are as set forth in SEQ ID NOs:53-55, and / or wherein the antibody comprises a light chain variable region comprising an amino acid sequence as set forth in SEQ ID NO:60, ii) an amino acid sequence with at least 80%, at least 90%, or at least 95% sequence identity to SEQ ID NO:60, wherein the CDR sequences are as set forth in SEQ ID NOs: 56-58.

36. An antibody comprising a light chain variable region and a heavy chain variable region, the heavy chain variable region comprising complimentary determining regions CDR-H1 , CDR- H2 and CDR-H3, the light chain variable region comprising complementarity determining regions CDR-L1, CDR-L2 and CDR-L3, wherein the antibody comprises a heavy chain variable region comprising: i) an amino acid sequence as set forth in SEQ ID NO: 67, ii) an amino acid sequence with at least 80%, at least 90%, or at least 95% sequence identity to SEQ ID NO: 67, wherein the CDR sequences are as set forth in SEQ ID NOs:61-63, and / or wherein the antibody comprises a light chain variable region comprising an amino acid sequence as set forth in SEQ ID NO: 68, ii) an amino acid sequence with at least 80%, at least 90%, or at least 95% sequence identity to SEQ ID NO:68, wherein the CDR sequences are as set forth in SEQ ID NOs: 64-66.

37. The antibody of claim 31 , wherein the antibody comprises a heavy chain variable region comprising: i) an amino acid sequence as set forth in SEQ ID NO: 75, ii) an amino acid sequence with at least 80%, at least 90%, or at least 95% sequence identity to SEQ ID NO: 75, wherein the CDR sequences are as set forth in SEQ ID NOs: 69-71 , and / or wherein theantibody comprises a light chain variable region comprising an amino acid sequence as set forth in SEQ ID NO: 76, ii) an amino acid sequence with at least 80%, at least 90%, or at least 95% sequence identity to SEQ ID NO:76, wherein the CDR sequences are as set forth in SEQ ID NOs: 72-74.

38. The antibody of claim 31 , wherein the heavy chain and the light chain have the sequences:Isotype SEQ ID NOs:Heavy 77 79 81 83 85 88 orLight 78 80 82 84 86 8939. The antibody of any one of claims 31 to 38, wherein the antibody is a single chain antibody or a binding fragment thereof.

40. The antibody of any one of claims 31 to 39, wherein the antibody is for use as or for the preparation of, a medicament.41 . A composition comprising the antibody of any one of claims 31 to 40.

42. The composition of claim 41 wherein the composition further comprises a pharmaceutically acceptable carrier.

43. The composition of claim 41 or 42, wherein the composition is formulated for IV or for subcutaneous administration.

Images