Anti-CB1 antibody compositions and therapeutic conjugates thereof

Abstract

The invention relates to therapeutic polypeptides and conjugates combining therapeutic molecules with anti-cannabinoid receptor 1 (CB1) antibodies and antigen-binding fragments thereof for the treatment, prevention, and / or amelioration of obesity and diabetes.

Description

MBHB Ref. No.: 24-0691 -WOANTI-CB1 ANTIBODY COMPOSITIONS AND THERAPEUTIC CONJUGATES THEREOFREFERENCE TO AN ELECTRONIC SEQUENCE LISTING

[0001] The instant application contains a Sequence Listing that has been submitted electronically and is hereby incorporated by reference in its entirety. The Sequence Listing was created on December 15, 2025, is named "24-0691-WO_SequenceListing.xml", and is 427,576 bytes in size.BACKGROUNDField of the Disclosure

[0002] The disclosure relates to therapeutic polypeptides and conjugates combining therapeutic molecules with anti-cannabinoid receptor 1 (CB1) antibodies and antigen-binding fragments thereof.Description of Related Art

[0003] Cannabinoid receptor 1 (CB1) is a G protein-coupled receptor (GPCR) expressed in the central nervous system (CNS), lungs, liver, adipose tissue and kidneys. It has been implicated in a range of human diseases, including obesity, diabetes, fibrosis, liver diseases, cardiovascular disease, cancer, pain, multiple sclerosis (MS) spasticity, and glaucoma. Specifically, CB1 activation has been shown to exert detrimental effects in conditions such as obesity, diabetes, fibrosis, liver diseases, cardiovascular disease and cancer, whereas it can exhibit beneficial activity in others, including pain, MS spasticity and glaucoma. Although several CB1-targeting compounds, including small molecules, and anti-CB1 antibodies, have been described, they are often associated with undesirable side effects. Thus, there remains a need in the art for strategies that mitigate side effects while enhancing therapeutic efficacy.SUMMARY

[0004] It is against the above background that the present invention provides certain advantages and advancements over prior art.MBHB Ref. No.: 24-0691 -WO

[0005] Although this invention as described herein is not limited to specific advantages or functionalities (such for example, therapeutic polypeptides comprising at least one non-proteinogenic residue and further comprising a prosthetic group, which can be conjugated to a proteinogenic or non-proteinogenic residue by a linker or spacer, and conjugates combining therapeutic polypeptides with anti-CB1 antibodies and antigen-binding fragments thereof), the invention provides an isolated peptide, comprising the amino acid sequence:

[0006] Xaa 1 -Xaa2-Xaa3-Xaa4-Xaa5-Xaa6-Xaa7-Xaa8-Xaa9-Xaa 10-Xaa 11 -Xaa 12-Xaa 13-Xaa14-Xaa15-Xaa16-Xaa17-Xaa18-Xaa19-Xaa20-Xaa21 -Xaa22-Xaa23-Xaa24-Xaa25-Xaa26-Xaa27-Xaa28-Xaa29-Xaa30-Xaa31-Xaa32-Xaa33-Xaa34-Xaa35-Xaa36-Xaa37-Xaa38-Xaa39-Xaa40 (SEQ ID NO: 1);wherein:Xaa1 is His, 3-Mal-PEGo-2o-Lys, 3-Mal-Lys, Tyr, pyroGlu, or 3-Mal-PEGo-2o;Xaa2 is Aib, Gly, Cys, Ser, Arg, or hArg;Xaa3 is Glu, Asn, Gin, Pro, or D-Arg;Xaa4 is Gly, Thr, Arg, or Gin;Xaa5 is Thr, Ala, Leu, or hArg;Xaa6 is Phe, Thr, Ser, or Pro;Xaa7 is Thr, Cys, His, or D-Arg;Xaa8 is Ser, Ala, £-3-Mal-PEGo-2o-Lys, or NMe-Leu;Xaa9 is Asp, Thr, Gly, or Ser;Xaa10 is Vai, Glu, Tyr, E-3-Mal-PEGo-2o-Lys, Pro, or His;Xaa11 is Ser, Arg, Met, or Lys;Xaa12 is Ser, Leu, lie, Glu, Lys, Pro, or Gly;Xaa13 is Tyr, Ala, Aib, Phe, or Oic;Xaa14 is Leu, Glu, NH, or pl-Phe;Xaa15 is Glu, Phe, Asp, Pro, or is absent;Xaa16 is Aib, Gly, Glu, Leu, Lys, Ser, D-Bip, or is absent;Xaa17 is Gin, Arg, lie, Glu, OH, or is absent;MBHB Ref. No.: 24-0691 -WOXaa18 is Ala, His, Arg, or is absent;Xaa19 is Ala, Ser, Gin, or is absent;Xaa20 is Lys, Ser, £-3-Mal-PEGo-2o-Lys, Arg, or is absent;Xaa21 is Glu, Asn, Ala, Asp, or is absent;Xaa22 is Phe, Asn, or is absent;Xaa23 is lie, Phe, Vai, or is absent;Xaa24 is Ala, Gly, Gin, Ala, Glu, or is absent;Xaa25 is Trp, Pro, Thr, or is absent;Xaa26 is Leu, lie, or is absent;Xaa27 is Vai, Leu, lie, Glu, or is absent;Xaa28 is Lys, Pro, Ala, Asn, or is absent;Xaa29 is Gly, Thr, Pro, or is absent;Xaa30 is Gly, Arg, Asn, Thr, £-BrAc-PEGo-2o-Lys, PEGo-2o-[£-3-Mal-D-Lys], or is absent;Xaa31 is Gly, (Sar)2.2o-[£-3-Mal-Lys], Vai, Asn, Pro, OH, NH2, or is absent;Xaa32 is (Gly-Gly-Gly-Gly-Ser)3-[£-BrAc-Lys], PEGo-2o-(£-3-Mal-Lys), NH2, (Sar)2.2o-[£-3-Mal-Lys], Gly, Vai, Ser, or is absent;Xaa33 is NH2, Ser, Gly, or is absent;Xaa34 is Asn, Ser, Gly, or is absent;Xaa35 is Thr, Asn, Ala, or is absent;Xaa36 is Pro, Thr, or is absent;Xaa37 is NH2, Pro, or is absent;Xaa38 is NH2, Pro, or is absent;Xaa39 is Ser or is absent; andXaa40 is NH2or is absent.

[0007] In some aspects of the peptides disclosed herein, Xaa1 is His, Xaa2 is Aib, Xaa3 is Glu, Xaa4 is Gly, Xaa5 is Thr, Xaa6 is Phe, Xaa7 is Thr, Xaa8 is Ser, Xaa9 is Asp, Xaa10 is Vai, Xaa11 is Ser, Xaa12 is Ser, Xaa13 is Tyr, Xaa14 is Leu, Xaa15 is Glu, Xaa16 is Aib, Xaa17 isMBHB Ref. No.: 24-0691 -WO

[0008] In some aspects of the peptides disclosed herein, Xaa1 is His, Xaa2 is Aib, Xaa3 is Glu, Xaa4 is Gly, Xaa5 is Thr, Xaa6 is Phe, Xaa7 is Thr, Xaa8 is Ser, Xaa9 is Asp, Xaa10 is Vai, Xaa11 is Ser, Xaa12 is Ser, Xaa13 is Tyr, Xaa14 is Leu, Xaa15 is Glu, Xaa16 is Aib, Xaa17 is Gin, Xaa18 is Ala, Xaa19 is Ala, Xaa20 is Lys, Xaa21 is Glu, Xaa22 is Phe, Xaa23 is He, Xaa24 is Ala, Xaa25 is Trp, Xaa26 is Leu, Xaa27 is Vai, Xaa28 is Lys, Xaa29 is Gly, Xaa30 is Gly, Xaa31 is Gly, Xaa32 is PEGi6-(£-3-Mal-Lys), Xaa33 is NH2, and Xaa34-Xaa40 are absent (SEQ ID NO: 3). In some aspects of the peptides disclosed herein, Xaa1 is His, Xaa2 is Gly, Xaa3 is Glu, Xaa4 is Gly, Xaa5 is Thr, Xaa6 is Phe, Xaa7 is Thr, Xaa8 is Ser, Xaa9 is Asp, Xaa10 is Vai, Xaa11 is Ser, Xaa12 is Ser, Xaa13 is Tyr, Xaa14 is Leu, Xaa15 is Glu, Xaa16 is Gly, Xaa17 is Gin, Xaa18 is Ala, Xaa19 is Ala, Xaa20 is Lys, Xaa21 is Glu, Xaa22 is Phe, Xaa23 is lie, Xaa24 is Ala, Xaa25 is Trp, Xaa26 is Leu, Xaa27 is Vai, Xaa28 is Lys, Xaa29 is Gly, Xaa30 is Arg, Xaa31 is (Sar)i6-[£-3-Mal-Lys], Xaa32 is NH2, and Xaa33-Xaa40 are absent (SEQ ID NO: 4).

[0009] In some aspects of the peptides disclosed herein, Xaa1 is His, Xaa2 is Gly, Xaa3 is Glu, Xaa4 is Gly, Xaa5 is Thr, Xaa6 is Phe, Xaa7 is Thr, Xaa8 is Ser, Xaa9 is Asp, Xaa10 is Vai, Xaa11 is Ser, Xaa12 is Ser, Xaa13 is Tyr, Xaa14 is Leu, Xaa15 is Glu, Xaa16 is Glu, Xaa17 is Gin, Xaa18 is Ala, Xaa19 is Ala, Xaa20 is Lys, Xaa21 is Glu, Xaa22 is Phe, Xaa23 is lie, Xaa24 is Ala, Xaa25 is Trp, Xaa26 is Leu, Xaa27 is Vai, Xaa28 is Lys, Xaa29 is Gly, Xaa30 is Gly, Xaa31 is Gly, Xaa32 is (Sar)i6-[£-3-Mal-Lys], Xaa33 is NH2, and Xaa34-Xaa40 are absent (SEQ ID NO: 5).

[0010] In some aspects of the peptides disclosed herein, Xaa1 is 3-Mal-PEGs-Lys, Xaa2 is Cys, Xaa3 is Asn, Xaa4 is Thr, Xaa5 is Ala, Xaa6 is Thr, Xaa7 is Cys, Xaa8 is Ala, Xaa9 is Thr, Xaa10 is Gin, Xaa11 is Arg, Xaa12 is Leu, Xaa13 is Ala, Xaa14 is Glu, Xaa15 is Phe, Xaa16 is Leu, Xaa17 is Arg, Xaa18 is His, Xaa19 is Ser, Xaa20 is Ser, Xaa21 is Asn, Xaa22 is Asn, Xaa23 is Phe, Xaa24 is Gly, Xaa25 is Pro, Xaa26 is lie, Xaa27 is Leu, Xaa28 is Pro, Xaa29 is Pro, Xaa30 is Thr, Xaa31 is Asn, Xaa32 is Vai, Xaa33 is Gly, Xaa34 is Ser, Xaa35 is Asn, Xaa36 is Thr, Xaa37 is Pro, Xaa38 is NH2, Xaa39-Xaa40 are absent, wherein the peptide further contains a disulfide bridge, Cys2-Cys7 (SEQ ID NO: 6).

[0011] In some aspects of the peptides disclosed herein, Xaa1 is 6-Mal-Lys, Xaa2 is Cys, Xaa3 is Asn, Xaa4 is Thr, Xaa5 is Ala, Xaa6 is Thr, Xaa7 is Cys, Xaa8 is Ala, Xaa9 is Thr, Xaa10MBHB Ref. No.: 24-0691 -WOis Gin, Xaa11 is Arg, Xaa12 is Leu, Xaa13 is Ala, Xaa14 is Glu, Xaa15 is Phe, Xaa16 is Leu, Xaa17 is Arg, Xaa18 is His, Xaa19 is Ser, Xaa20 is Ser, Xaa21 is Asn, Xaa22 is Asn, Xaa23 is Phe, Xaa24 is Gly, Xaa25 is Pro, Xaa26 is lie, Xaa27 is Leu, Xaa28 is Pro, Xaa29 is Pro, Xaa30 is Thr, Xaa31 is Asn, Xaa32 is Vai, Xaa33 is Gly, Xaa34 is Ser, Xaa35 is Asn, Xaa36 is Thr, Xaa37 is Pro, Xaa38 is NH2, Xaa39-Xaa40 are absent, wherein the peptide further contains a disulfide bridge, Cys2-Cys7 (SEQ ID NO: 7).

[0012] In some aspects of the peptides disclosed herein, Xaa1 is Tyr, Xaa2 is Aib, Xaa3 is Glu, Xaa4 is Gly, Xaa5 is Thr, Xaa6 is Phe, Xaa7 is Thr, Xaa8 is Ser, Xaa9 is Asp, Xaa10 is Tyr, Xaa11 is Ser, Xaa12 is lie, Xaa13 is Aib, Xaa14 is Leu, Xaa15 is Asp, Xaa16 is Lys, Xaa17 is lie, Xaa18 is Ala, Xaa19 is Gin, Xaa20 is e-3-Mal-PEGs-Lys, Xaa21 is Ala, Xaa22 is Phe, Xaa23 is Vai, Xaa24 is Gin, Xaa25 is Trp, Xaa26 is Leu, Xaa27 is lie, Xaa28 is Ala, Xaa29 is Gly, Xaa30 is Gly, Xaa31 is Pro, Xaa32 is Ser, Xaa33 is Ser, Xaa34 is Gly, Xaa35 is Ala, Xaa36 is Pro, Xaa37 is Pro, Xaa38 is Pro, Xaa39 is Ser, Xaa40 is NH2(SEQ ID NO: 8).

[0013] In some aspects of the peptides disclosed herein, Xaa1 is His, Xaa2 is Ser, Xaa3 is Gin, Xaa4 is Gly, Xaa5 is Thr, Xaa6 is Phe, Xaa7 is Thr, Xaa8 is Ser, Xaa9 is Asp, Xaa10 is £-3-Mal-PEGs-Lys, Xaa11 is Ser, Xaa12 is Glu, Xaa13 is Tyr, Xaa14 is Leu, Xaa15 is Asp, Xaa16 is Ser, Xaa17 is Glu, Xaa18 is Arg, Xaa19 is Ala, Xaa20 is Arg, Xaa21 is Asp, Xaa22 is Phe, Xaa23 is Vai, Xaa24 is Ala, Xaa25 is Trp, Xaa26 is Leu, Xaa27 is Glu, Xaa28 is Ala, Xaa29 is Gly, Xaa30 is Gly, Xaa31 is OH, and Xaa32-Xaa40 are absent (SEQ ID NO: 9).

[0014] In some aspects of the peptides disclosed herein, Xaa1 is His, Xaa2 is Aib, Xaa3 is Gin, Xaa4 is Gly, Xaa5 is Thr, Xaa6 is Phe, Xaa7 is Thr, Xaa8 is Ser, Xaa9 is Asp, Xaa10 is Tyr, Xaa11 is Ser, Xaa12 is Lys, Xaa13 is Tyr, Xaa14 is Leu, Xaa15 is Asp, Xaa16 is Glu, Xaa17 is Arg, Xaa18 is Arg, Xaa19 is Ala, Xaa20 is Arg, Xaa21 is Asp, Xaa22 is Phe, Xaa23 is Vai, Xaa24 is Glu, Xaa25 is Trp, Xaa26 is Leu, Xaa27 is Leu, Xaa28 is Asn, Xaa29 is Thr, Xaa30 is £-BrAc-PEGi2-Lys, Xaa31 is NH2, and Xaa32-Xaa40 are absent (SEQ ID NO: 10).

[0015] In some aspects of the peptides disclosed herein, Xaa1 is His, Xaa2 is Aib, Xaa3 is Gin, Xaa4 is Gly, Xaa5 is Thr, Xaa6 is Phe, Xaa7 is Thr, Xaa8 is Ser, Xaa9 is Asp, Xaa10 is Tyr, Xaa11 is Ser, Xaa12 is Lys, Xaa13 is Tyr, Xaa14 is Leu, Xaa15 is Asp, Xaa16 is Glu, Xaa17 is Arg, Xaa18 is Arg, Xaa19 is Ala, Xaa20 is Arg, Xaa21 is Asp, Xaa22 is Phe, Xaa23 is Vai, Xaa24 is Glu, Xaa25 is Trp, Xaa26 is Leu, Xaa27 is Leu, Xaa28 is Asn, Xaa29 is Thr, Xaa30 is PEGs-[£-3-Mal-D-Lys], Xaa31 is NH2, and Xaa32-Xaa40 are absent (SEQ ID NO: 11).MBHB Ref. No.: 24-0691 -WO

[0016] In some aspects of the peptides disclosed herein, Xaa1 is pyroGlu, Xaa2 is Arg, Xaa3 is Pro, Xaa4 is Arg, Xaa5 is Leu, Xaa6 is Ser, Xaa7 is His, Xaa8 is e-3-Mal-PEGs-Lys, Xaa9 is Gly, Xaa10 is Pro, Xaa11 is Met, Xaa12 is Pro, Xaa13 is Phe, Xaa14 NH, and Xaa15-Xaa40 are absent (SEQ ID NO: 12).

[0017] In some aspects of the peptides disclosed herein, Xaa1 is 3-Mal-PEG8, Xaa2 is hArg, Xaa3 is D-Arg, Xaa4 is Gin, Xaa5 is hArg, Xaa6 is Pro, Xaa7 is D-Arg, Xaa8 is NMe-Leu, Xaa9 is Ser, Xaa10 is His, Xaa11 is Lys, Xaa12 is Gly, Xaa13 is Oic, Xaa14 is pl-Phe, Xaa15 is Pro, Xaa16 is D-Bip, Xaa17 is OH, and Xaa18-Xaa40 are absent (SEQ ID NO: 13).

[0018] In some aspects of the peptides disclosed herein, the peptide has the amino acid sequence:(a) His-Aib-Glu-Gly-Thr-Phe-Thr-Ser-Asp-Val-Ser-Ser-Tyr-Leu-Glu-Aib-GIn-Ala-Ala-Lys- Glu-Phe-lle-Ala-Trp-Leu-Val-Lys-Gly-Gly-Gly-(Gly-Gly-Gly-Gly-Ser)3-[£-BrAc-Lys]-NH2(SEQ ID NO: 2), wherein [e-BrAc-Lys]-NH2has the following structure(b) His-Aib-Glu-Gly-Thr-Phe-Thr-Ser-Asp-Val-Ser-Ser-Tyr-Leu-Glu-Aib-GIn-Ala-Ala-Lys- Glu-Phe-lle-Ala-Trp-Leu-Val-Lys-Gly-Gly-Gly-[PEGi6-(£-3-Mal-Lys)]-NH2(SEQ ID NO: 3), wherein [PEGi6-(£-3-Mal-Lys)]-NH2has the following structure:(c) His-Gly-Glu-Gly-Thr-Phe-Thr-Ser-Asp-Val-Ser-Ser-Tyr-Leu-Glu-Gly-GIn-Ala-Ala-Lys- Glu-Phe-lle-Ala-Trp-Leu-Val-Lys-Gly-Arg-(Sar)i6-[£-3-Mal-Lys]-NH2(SEQ ID NO: 4), wherein -[£- 3-Mal-Lys]-NH2has the following structure:MBHB Ref. No.: 24-0691 -WOArg-His-Ser-Ser-Asn-Asn-Phe-Gly-Pro-lle-Leu-Pro-Pro-Thr-Asn-Val-Gly-Ser-Asn-Thr-Pro-NH2(disulfide bridge, Cys2-Cys7) (SEQ ID NO: 6), wherein [3-Mal-PEGs-Lys] has the following structure:-Ala-Glu-Phe-Leu-Arg- His-Ser-Ser-Asn-Asn-Phe-Gly-Pro-lle-Leu-Pro-Pro-Thr-Asn-Val-Gly-Ser-Asn-Thr-Pro-NH2(disulfide bridge, Cys2-Cys7) (SEQ ID NO: 7), wherein [6-Mal-Lys]- has the following structure:MBHB Ref. No.: 24-0691 -WOMBHB Ref. No.: 24-0691 -WO(k) [pyroGlu]-Arg-Pro-Arg-Leu-Ser-His-[£-3-Mal-PEG8-Lys]-Gly-Pro-Met-Pro-Phe-NH2(SEQ ID NO: 12), wherein [3-Mal-PEG8] has the following structure:wherein the complete structural formula is as follows:

[0019] The invention also provides a pharmaceutical composition, comprising the peptides disclosed herein and a pharmaceutically acceptable excipient, carrier, or diluent.MBHB Ref. No.: 24-0691 -WO

[0020] The invention also provides a method for treating obesity and / or type 2 diabetes in a subject, the method comprising administering to the subject in need of such treatment an effective amount of the pharmaceutical composition disclosed herein.

[0021] The invention also provides a kit, comprising the peptides or the pharmaceutical compositions disclosed herein.

[0022] The invention also provides a conjugate, comprising:(a) at least one of the peptides disclosed herein; and(b) an isolated antibody or an antigen binding fragment thereof that specifically binds to human cannabinoid receptor 1 (CB1).In some embodiments, the isolated antibody or antigen binding fragment thereof comprises: (i) a heavy chain variable (VH) region comprising the following complementarity determining regions (CDRs):HCDR1 having the amino acid sequence of SEQ ID NO: 26;HCDR2 having the amino acid sequence of SEQ ID NO: 27;HCDR3 having the amino acid sequence of SEQ ID NO: 28; and(ii) a light chain variable (VL) region comprising the following CDRs:LCDR1 having the amino acid sequence of SEQ ID NO: 29;LCDR2 having the amino acid sequence of SEQ ID NO: 30;LCDR3 having the amino acid sequence of SEQ ID NO: 31; oror a pharmaceutically acceptable salt, solvate, stereoisomer, or tautomer thereof.

[0023] In some aspects of the conjugates disclosed herein, the VH region of the isolated antibody or the antigen binding fragment thereof comprises an amino acid sequence that is 90% identical to SEQ ID NO: 32.

[0024] In some aspects of the conjugates disclosed herein, the VL region of the isolated antibody or the antigen binding fragment thereof comprises an amino acid sequence that is 90% identical to SEQ ID NO: 33.

[0025] In some aspects of the conjugates disclosed herein, the isolated antibody or the antigen binding fragment thereof comprises:MBHB Ref. No.: 24-0691 -WO(a) a heavy chain comprising an amino acid sequence at least 90% identical to SEQ ID NO: 32; and(b) a light chain variable region comprising an amino acid sequence that is at least 90% identical to SEQ ID NO: 33.

[0026] In some aspects of the conjugates disclosed herein, the conjugate has a drug-antibody ratio of the at least one of the peptides of any one of claims 1-4 to the isolated antibody or the antigen binding fragment thereof. In some embodiments, the drug-antibody ratio is greater than 1. In some embodiments, the drug-antibody ratio is about 2. In some embodiments, the drugantibody ratio is about 4.

[0027] In some aspects of the conjugates disclosed herein, the at least one of the peptides disclosed herein is linked to the isolated antibody or the antigen binding fragment thereof via cysteine-based bioconjugation.

[0028] In some aspects of the conjugates disclosed herein, the at least one of the peptides disclosed herein is linked site-specifically to the isolated antibody or the antigen binding fragment thereof.

[0029] In some aspects of the conjugates disclosed herein, the at least one of the peptides disclosed herein is conjugated to the N-terminus of the isolated antibody or the antigen binding fragment.

[0030] In some aspects of the conjugates disclosed herein, the at least one of the peptides disclosed herein is conjugated to the C-terminus of the isolated antibody or the antigen binding fragment.

[0031] In some aspects of the conjugates disclosed herein, the at least one of the peptides disclosed herein is conjugated to the Fc region of the isolated antibody or the antigen binding fragment.

[0032] The invention also provides a pharmaceutical composition, comprising the conjugates disclosed herein and a pharmaceutically acceptable excipient, carrier, or diluent.

[0033] The invention also provides a method for treating obesity and / or type 2 diabetes in a subject, the method comprising administering to the subject in need of such treatment an effective amount the pharmaceutical composition disclosed herein.

[0034] The invention also provides a kit, comprising the conjugates or the pharmaceutical composition disclosed herein.MBHB Ref. No.: 24-0691 -WO

[0035] These and other features and advantages of the present invention will be more fully understood from the following detailed description taken together with the accompanying claims. It is noted that the scope of the claims is defined by the recitations therein and not by the specific discussion of features and advantages set forth in the present descriptionBRIEF DESCRIPTION OF THE DRAWINGS

[0036] The following detailed description of the embodiments of the present invention can be best understood when read in conjunction with the following drawings, where like structure is indicated with like reference numerals and in which:

[0037] FIG. 1 shows disulfide bonds in a full IgG scaffold.

[0038] FIG. 2 shows a scheme of cysteine-based bioconjugation of a monoclonal antibody with SBI-407, a maleimide-containing peptide disclosed herein. It should be understood that the DAR value and linkage sites may vary, as disclosed herein.

[0039] FIG. 3 shows a scheme of cysteine-based bioconjugation of a monoclonal antibody with SBI-401, an c-BrAc-Lys containing peptide disclosed herein. It should be understood that the DAR value and linkage sites may vary, as disclosed herein.

[0040] FIG. 4 shows potential isomers of DAR2 (E2A, E2B), DAR4 (E4A, E4B, E4C) and DAR6 (E6A, E6B). The locations of conjugation are indicated by stars and intact disulfide bonds are shown as bars (Sun et al., 2005, Bioconjugate Chem. 16(5): 1282-1290).

[0041] FIGS. 5A-5L show HPLC run information and chromatograms for each peptide. FIG.5A depicts information for SBI-401 , FIG. 5B depicts information for SBI-402, FIG. 5C depicts information for SBI-403, FIG. 5D depicts information for SBI-404, FIG. 5E depicts information for SBI-405, FIG. 5F depicts information for SBI-406, FIG. 5G depicts information for SBI-407, FIG.5H depicts information for SBI-408, FIG. 5I depicts information for SBI-409, FIG. 5J depicts information forSBI-410, FIG. 5K depicts information for SBI-411, and FIG.5L depicts information for SBI-412.

[0042] FIGS. 6A-6L show mass spectra and identified adducts for each peptide. FIG. 6A depicts information for SBI-401, FIG. 6B depicts information for SBI-402, FIG. 6C depicts information for SBI-403, FIG. 6D depicts information for SBI-404, FIG. 6E depicts information for SBI-405, FIG. 6F depicts information for SBI-406, FIG. 6G depicts information for SBI-407, FIG.6H depicts information for SBI-408, FIG. 6I depicts information for SBI-409, FIG. 6J depictsMBHB Ref. No.: 24-0691 -WOinformation for SBI-410, FIG. 6K depicts information for SBI-411, and FIG. 6L depicts information forSBI-412.

[0043] FIGS. 7A-7J show CB1 inhibition assays for P1C4-peptide conjugates with DAR of 2 (“D2”) and 4 (“D4”) compared to nimacimab. RLU, relative light units; EC5o, half-maximal effective concentration. FIG. 7A shows P1C4-401-D2 and P1C4-401-D4. FIG. 7B shows P1C4-403-D2 and P1C4-403-D4. FIG. 7C shows P1C4-404-D2 and P1C4-404-D4. FIG. 7D shows P1C4-405-D2 and P1C4-405-D4. FIG. 7E shows P1C4-406-D2 and P1C4-406-D4. FIG. 7F shows P1C4-407-D2 and P1C4-407-D4. FIG. 7G shows P1C4-408-D2 and P1C4-408-D4. FIG. 7H shows P1C4-409-D2 and P1C4-409-D4. FIG. 7I shows P1C4-410-D2 and P1C4-410-D4. FIG. 7J shows P1C4-411-D2 and P1C4-411-D4.

[0044] FIGS. 8A-8G show GLP-1 receptor activation assays for selected P1C4-peptide conjugates compared to nimacimab, corresponding peptide alone (SBI-401, SBI-403, SBI-404, SBI-407, SBI-408, SBI-409, and SBI-410), and GLP-1, residues 7-36. RLU, relative light units; ECso, half-maximal effective concentration. RLU are normalized. FIG. 8A shows P1C4-401-D2 and P1C4-401-D4. FIG. 8B shows P1C4-403-D2 and P1C4-403-D4. FIG. 8C shows P1C4-404-D2 and P1C4-404-D4. FIG. 8D shows P1C4-407-D2 and P1C4-407-D4 with an additional tirzepatide control. FIG. 8E shows P1C4-408-D2 and P1C4-408-D4 with an additional tirzepatide control. FIG. 8F shows P1C4-409-D2 and P1C4-409-D4 with an additional tirzepatide control.FIG. 8G shows P1C4-410-D2 and P1C4-410-D4 with an additional tirzepatide control.

[0045] FIGS. 9A-9G show GLP-1 receptor activation assays for selected P1C4-peptide conjugates compared to nimacimab, corresponding peptide alone (SBI-401 , SBI-403, SBI-404, SBI-407, SBI-408, SBI-409, and SBI-410), tirzepatide, and GLP-1. RLU, relative light units; EC50, half-maximal effective concentration. RLU are not normalized. FIG. 9A shows P1C4-401-D2 and P1C4-401-D4. FIG. 9B shows P1C4-403-D2 and P1C4-403-D4. FIG. 9C shows P1C4-404-D2 and P1C4-404-D4. FIG. 9D shows P1C4-407-D2 and P1C4-407-D4. FIG. 9E shows P1C4-408-D2 and P1C4-408-D4. FIG. 9F shows P1C4-409-D2 and P1C4-409-D4. FIG. 9G shows P1C4-410-D2 and P1C4-410-D4.

[0046] FIGS. 10A-10C show GLP-1 receptor activation assays of all GLP-1 R targeting moieties, accompanied by tirzepatide and GLP-1 controls. RLU, relative light units. FIG. 10A shows data from the peptides alone. FIG. 10B shows data from the conjugates with a DAR of 2.FIG. 10C shows data from the conjugates with a DAR of 4.

[0047] FIGS. 11A-11C show the data from FIGS. 10A-10C with RLU normalized.MBHB Ref. No.: 24-0691 -WO

[0048] FIGS. 12A-12B show amylin receptor activation assays for selected P1C4-peptide conjugates alongside amylin control and corresponding peptide alone (SBI-405 and SBI-406).FIG. 12A shows P1C4-405-D2 and P1C4-405-D4. FIG. 12B shows P1C4-406-D2 and P1C4-406-D4.

[0049] FIG. 13 shows a GIP receptor activation assay for selected P1C4-peptide conjugates alongside nimacimab, corresponding peptide alone (SBI-407), GIP and tirzepatide. P1C4-407-D2, conjugate with DAR 2; P1C4-407-D4, conjugate with DAR 4. RLU are normalized.

[0050] FIG. 14 shows GCGR (glucagon receptor) activation assays for corresponding peptide alone (SBI-408, SBI-409, SBI-410) alongside glucagon control.

[0051] FIGS. 15A-15C show GCGR activation assays for selected P1C4-peptide conjugates compared to nimacimab, corresponding peptide alone (SBI-408, SBI-409, and SBI-410), glucagon, and retatrutide. FIG. 15A shows P1C4-408-D2 and P1C4-408-D4. FIG. 15B shows P1C4-409-D2 and P1C4-409-D4. FIG. 15C shows P1C4-410-D2 and P1C4-410-D4.

[0052] FIG. 16 shows an apelin receptor activation assay for the SBI-411 series peptide and conjugates alongside apelin-13 control. SBI-411, peptide alone; P1C4-411-D2, conjugate with DAR 2; P1C4-411-D4, conjugate with DAR4.

[0053] Skilled artisans will appreciate that elements in the drawings are illustrated for simplicity and clarity and have not necessarily been drawn to scale. For example, the dimensions of some of the elements in the Figures can be exaggerated relative to other elements to help improve understanding of the embodiment(s) of the present invention.DETAILED DESCRIPTION OF THE INVENTION

[0054] It is to be understood that the particular aspects of the specification are described herein are not limited to specific embodiments presented and can vary. It also will be understood that the terminology used herein is for the purpose of describing particular aspects only and, unless specifically defined herein, is not intended to be limiting. Moreover, particular embodiments disclosed herein can be combined with other embodiments disclosed herein, as would be recognized by a skilled person, without limitation.

[0055] All publications, including but not limited to journal articles, patents, and patent applications, cited in this specification are herein incorporated by reference as though set forth in their entirety in the present application.MBHB Ref. No.: 24-0691 -WODefinitions

[0056] Before describing the methods and compositions of the disclosure in detail, a number of terms will be defined. As used herein, the singular forms “a,” “an,” and “the” include plural referents unless the context clearly dictates otherwise. For example, reference to “a peptide” means one or more peptide molecules.

[0057] Throughout this specification, unless the context specifically indicates otherwise, the terms “comprise” and “include” and variations thereof (e.g., “comprises,” “comprising,” “includes,” and “including”) will be understood to indicate the inclusion of a stated component, feature, element, or step or group of components, features, elements or steps but not the exclusion of any other component, feature, element, or step or group of components, features, elements, or steps. Any of the terms “comprising,” “consisting essentially of,” and “consisting of’ may be replaced with either of the other two terms, while retaining their ordinary meanings.

[0058] In some embodiments, percentages disclosed herein can vary in amount by ±10, 20, or 30% from values disclosed and remain within the scope of the contemplated disclosure.

[0059] Unless otherwise indicated or otherwise evident from the context and understanding of one of ordinary skill in the art, values herein that are expressed as ranges can assume any specific value or sub-range within the stated ranges in different embodiments of the disclosure, to the tenth of the unit of the lower limit of the range, unless the context clearly dictates otherwise.

[0060] As used herein, ranges and amounts can be expressed as “about” a particular value or range. “About” also includes the exact amount. For example, “about 5%” means “about 5%” and also “5%”. The term “about” can also refer to ± 10% of a given value or range of values, or to a value on a similar order of magnitude. Therefore, about 5% also means 4.5% - 5.5%, for example, or 1-10%.

[0061] As used herein, the terms “or” and “and / or” are utilized to describe multiple components in combination or exclusive of one another. For example, “x, y, and / or z” can refer to “x” alone, “y” alone, “z” alone, “x, y, and z,” “(x and y) or z,” “x or (y and z) ,” or “x or y or z.”

[0062] It is noted that terms like “preferably,” “commonly,” and “typically” are not utilized herein to limit the scope of the methods and compositions as described herein or to imply that certain features are critical, essential, or even important to the structure or function of the subject matter recited in the claims.MBHB Ref. No.: 24-0691 -WO

[0063] As utilized in accordance with the present disclosure, unless otherwise indicated, all technical and scientific terms shall be understood to have the meaning commonly understood by one of ordinary skill in the art.

[0064] Abbreviations used herein include:MBHB Ref. No.: 24-0691 -WO

[0065] The term “polypeptide” and “peptide” as used herein means a compound containing at least two constituent amino acids connected by peptide bonds. As used herein, a peptide is defined as a molecule that contains two or more amino acids. Peptides that contain many amino acids are called polypeptides or proteins. Thus, as used herein, the terms “peptide” and “protein” are interchangeable. The choice of term makes no assertion regarding any particular properties of the molecule, including but not limited to length, molecular weight, or secondary structure.

[0066] Each constituent amino acid can be known as a “residue” of the peptide. Residues can be numbered, generally from the N-terminus of the peptide toward the C-terminus. The constituent amino acids may be from the group of the amino acids encoded by the genetic code and they may be natural amino acids which are not encoded by the genetic code, as well as synthetic amino acids. Natural amino acids which are not encoded by the genetic code are, for example, y-carboxyglutamate, ornithine, phosphoserine, D-alanine and D-glutamine. Synthetic amino acids comprise amino acids manufactured by chemical synthesis, i.e. D-isomers of the amino acids encoded by the genetic code such as D-alanine and D-leucine, Aib (a-aminoisobutyric acid), Abu (a-aminobutyric acid), Tie (tert-butylglycine), p-alanine, 3-aminomethyl benzoic acid, anthranilic acid, and others.

[0067] Amino acids encoded by the genetic code are also called “proteinogenic” amino acids. The 22 proteinogenic amino acids (and their one- and three-letter codes) are: alanine (A / Ala), arginine (R / Arg), asparagine (N / Asn), aspartic acid / aspartate (D / Asp), cysteine (C / Cys), cystine, glutamine (Q / GIn), glutamic acid / glutamate (E / Glu), glycine (G / Gly), histidine (H / His), hydroxyproline, isoleucine (l / lle), leucine (L / Leu), lysine (K / Lys), methionine (M / Met), phenylalanine (F / Phe), proline (P / Pro), serine (S / Ser), threonine (T / Thr), tryptophan (W / Trp), tyrosine (Y / Tr), and valine (V / Val).

[0068] A non-proteinogenic amino acid is a moiety which can be incorporated into a peptide via peptide bonds but is not a proteinogenic amino acid. Non-proteinogenic amino acids (NPAAs) are also known as “unnatural” amino acids or “non-naturally occurring” amino acids. Examples are y-carboxyglutamate, ornithine, phosphoserine, synthetic non-proteinogenic amino acids comprising amino acids manufactured by chemical synthesis, D-isomers of the amino acids encoded by the genetic code such as D-alanine and D-leucine, Aib (a-aminoisobutyric acid), Abu (a-aminobutyric acid), Tie (tert-butylglycine), 3-aminomethyl benzoic acid, anthranilic acid, desamino-histidine, the p-analogs of a-amino acids such as p-alanine, p-hydroxy-histidine, homohistidine, Na-acetyl-histidine, a-fluoromethyl-histidine, a-methyl-histidine, 3-pyridylalanine, 2-pyridylalanine or 4-pyridylalanine, (1 -aminocyclopropyl) carboxylic acid, (1-aminocyclobutyl)MBHB Ref. No.: 24-0691 -WOcarboxylic acid, (1 -aminocyclopentyl) carboxylic acid, (1 -aminocyclohexyl) carboxylic acid, (1-aminocycloheptyl) carboxylic acid, (1-aminocyclooctyl) carboxylic acid, £-(palmitoyl-y-Glu)-Lys, e-BrAc-PEGo-20-Lys, e-BrAc-Lys, PEGO-2o-(E-3-Mal-Lys), e-3-Mal-Lys, 3-Mal-PEGO-2o-Lys, 6-Mal-Lys, e-3-Mal-D-Lys, pyroglutamic acid (pyroGlu), L-homoarginine (hArg), D-arginine (D-Arg), / V-methyl-L-leucine (NMe-Leu), octahydroindole-2-carboxylic acid (Oic), 4-iodo-L-phenylalanine (pl-Phe), sarcosine (Sar), or (R)-3-([1,1'-biphenyl]-4-yl)-2-aminopropanoic acid (D-Bip).

[0069] The term “analog” as used herein referring to a polypeptide means a modified peptide wherein one or more amino acid residues of the peptide have been substituted by other amino acid residues, including non-proteinogenic amino acid residues, and / or wherein one or more amino acid residues have been deleted from the peptide and / or wherein one or more amino acid residues have been deleted from the peptide and / or wherein one or more amino acid residues have been added to the peptide. Such addition or deletion of amino acid residues can take place at the N-terminal of the peptide and / or at the C-terminal of the peptide. A simple system is often used to describe analogs; for example, [Arg34]GLP-1(7-37)Lys designates a GLP-1(7-37) analog wherein the naturally occurring lysine at position 34 has been substituted with arginine and wherein a lysine has been added to the terminal amino acid residue, i.e., to the Gly37. All amino acids for which the optical isomer is not stated is to be understood to mean the L-isomer.

[0070] The term “derivative” as used herein in relation to a peptide means a chemically modified peptide or an analog thereof, wherein at least one substituent is not present in the unmodified peptide or an analog thereof, i.e., a peptide where at least one residue has been covalently modified. Typical modifications are amides, carbohydrates, glycosyl groups, alkyl groups, acyl groups, esters, fatty acids, and the like. The process of making a peptide or amino acid derivative is called “derivatization.”Glucagon-like peptide 1 (GLP-1) and analogs

[0071] In this disclosure, peptide sequences are presented from N-terminus to C-terminus. When a peptide ends in “NH2”, this indicates that the peptide has been modified to end in an amide rather than the usual carboxylate. When a peptide as written ends in an explicit “OH”, this has an identical meaning to not explicitly including the “OH” — both indicate the usual carboxylate C-terminus.

[0072] Many glucagon-like peptide 1 (GLP-1) analogs have been developed for use as antiobesity and anti-diabetes drugs. These analogs are similar to portions of the native GLP-1 peptideMBHB Ref. No.: 24-0691 -WObut contain minor sequence variations and residue modifications. Some analogs consist of one or more copies of native GLP-1 conjugated to another native protein, such as human albumin. Such analogs include dulaglutide, AMG133 (Veniant et al., 2023, Nat. Metal. 6: 290-303), GLP-1 P1 (Lu et al., 2021, Cell Rep. Med. 2(5): 100263), semaglutide, albiglutide, exenatide, liraglutide, lixisenatide, and tirzepatide. These molecules exert agonist activity on GLP-1 receptors and thereby activate those receptors.

[0073] The sequence of residues 7-37 of the native GLP-1 peptide can be found below.

[0074] GLP-1 (7-37) (SEQ ID NO: 14)His-Ala-Glu-Gly-Thr-Phe-Thr-Ser-Asp-Val-Ser-Ser-Tyr-Leu-Glu-Gly-GIn-Ala-Ala-Lys-Glu-Phe-lle-Ala-Trp-Leu-Val-Lys-Gly-Arg-Gly-OH

[0075] Portions or subsets of the sequence of native GLP-1 can be modified to end in an amino group, as below.

[0076] GLP-1 (7-36) amide (SEQ ID NO: 15)His-Ala-Glu-Gly-Thr-Phe-Thr-Ser-Asp-Val-Ser-Ser-Tyr-Leu-Glu-Gly-GIn-Ala-Ala-Lys-Glu-Phe-lle-Ala-Trp-Leu-Val-Lys-Gly-Arg-NH2

[0077] Amino acid sequences of some examples of GLP-1 receptor agonists, which are analogs of GLP-1, can be found below.

[0078] Dulaglutide (GLP-1 (7-37)) (SEQ ID NO: 16)His-Gly-Glu-Gly-Thr-Phe-Thr-Ser-Asp-Val-Ser-Ser-Tyr-Leu-Glu-Glu-GIn-Ala-Ala-Lys-Glu-Phe-lle-Ala-Trp-Leu-Val-Lys-Gly-Gly-Gly-

[0079] AMG133 (SEQ ID NO: 17)His-Aib-Glu-Gly-Thr-Phe-Thr-Ser-Asp-Tyr-Ser-Ser-Tyr-Leu-Glu-Glu-GIn-Ala-Ala-Lys-Glu-Phe-lle-Ala-Trp-Leu-Val-Lys-Gly-Gly-Gly-

[0080] GLP-1 P1 (SEQ ID NO: 18)His-Aib-Glu-Gly-Thr-Phe-Thr-Ser-Asp-Val-Ser-Ser-Tyr-Leu-Glu-Aib-GIn-Ala-Ala-Lys-Glu-Phe-lle-Ala-Trp-Leu-Val-Lys-Gly-Gly-Gly-

[0081] Semaglutide (SEQ ID NO: 19)His-Aib-Glu-Gly-Thr-Phe-Thr-Ser-Asp-Val-Ser-Ser-Tyr-Leu-Glu-Gly-Gln-Ala-Ala-[C18 diacid-y-Glu-(AEEA)2-Lys]-Glu-Phe-lle-Ala-Trp-Leu-Val-Arg-Gly-Arg-Gly-OHMBHB Ref. No.: 24-0691 -WO

[0082] Albiglutide (SEQ ID NO: 20)His-Gly-Glu-Gly-Thr-Phe-Thr-Ser-Asp-Val-Ser-Ser-Tyr-Leu-Glu-Gly-GIn-Ala-Ala-Lys-Glu-Phe-lle-Ala-Trp-Leu-Val-Lys-Gly-Arg-His-Gly-Glu-Gly-Thr-Phe-Thr-Ser-Asp-Val-Ser-Ser-Tyr-Leu- Glu-Gly-GIn-Ala-Ala-Lys-Glu-Phe-lle-Ala-Trp-Leu-Val-Lys-Gly-Arg-His-Gly-Glu-Gly-Thr-Phe- Thr-Ser-Asp-Val-Ser-Ser-Tyr-Leu-Glu-Gly-GIn-Ala-Ala-Lys-Glu-Phe-lle-Ala-Trp-Leu-Val-Lys- Gly-Arg-Asp-Ala-His-Lys-Ser-Glu-Val-Ala-His-Arg-Phe-Lys-Asp-Leu-Gly-Glu-Glu-Asn-Phe-Lys- Ala-Leu-Val-Leu-lle-Ala-Phe-Ala-GIn-Tyr-Leu-GIn-GIn-Cys-Pro-Phe-Glu-Asp-His-Val-Lys-Leu- Val-Asn-Glu-Val-Thr-Glu-Phe-Ala-Lys-Thr-Cys-Val-Ala-Asp-Glu-Ser-Ala-Glu-Asn-Cys-Asp-Lys- Ser-Leu-His-Thr-Leu-Phe-Gly-Asp-Lys-Leu-Cys-Thr-Val-Ala-Thr-Leu-Arg-Glu-Thr-Tyr-Gly-Glu- Met-Ala-Asp-Cys-Cys-Ala-Lys-GIn-Glu-Pro-Glu-Arg-Asn-Glu-Cys-Phe-Leu-GIn-His-Lys-Asp- Asp-Asn-Pro-Asn-Leu-Pro-Arg-Leu-Val-Arg-Pro-Glu-Val-Asp-Val-Met-Cys-Thr-Ala-Phe-His-Asp- Asn-Glu-Glu-Thr-Phe-Leu-Lys-Lys-Tyr-Leu-Tyr-Glu-lle-Ala-Arg-Arg-His-Pro-Tyr-Phe-Tyr-Ala- Pro-Glu-Leu-Leu-Phe-Phe-Ala-Lys-Arg-Tyr-Lys-Ala-Ala-Phe-Thr-Glu-Cys-Cys-GIn-Ala-Ala-Asp- Lys-Ala-Ala-Cys-Leu-Leu-Pro-Lys-Leu-Asp-Glu-Leu-Arg-Asp-Glu-Gly-Lys-Ala-Ser-Ser-Ala-Lys- Gln-Arg-Leu-Lys-Cys-Ala-Ser-Leu-Gln-Lys-Phe-Gly-Glu-Arg-Ala-Phe-Lys-Ala-Trp-Ala-Val-Ala- Arg-Leu-Ser-GIn-Arg-Phe-Pro-Lys-Ala-Glu-Phe-Ala-Glu-Val-Ser-Lys-Leu-Val-Thr-Asp-Leu-Thr- Lys-Val-His-Thr-Glu-Cys-Cys-His-Gly-Asp-Leu-Leu-Glu-Cys-Ala-Asp-Asp-Arg-Ala-Asp-Leu-Ala- Lys-Tyr-lle-Cys-Glu-Asn-GIn-Asp-Ser-lle-Ser-Ser-Lys-Leu-Lys-Glu-Cys-Cys-Glu-Lys-Pro-Leu- Leu-Glu-Lys-Ser-His-Cys-lle-Ala-Glu-Val-Glu-Asn-Asp-Glu-Met-Pro-Ala-Asp-Leu-Pro-Ser-Leu- Ala-Ala-Asp-Phe-Val-Glu-Ser-Lys-Asp-Val-Cys-Lys-Asn-Tyr-Ala-Glu-Ala-Lys-Asp-Val-Phe-Leu- Gly-Met-Phe-Leu-Tyr-Glu-Tyr-Ala-Arg-Arg-His-Pro-Asp-Tyr-Ser-Val-Val-Leu-Leu-Leu-Arg-Leu- Ala-Lys-Thr-Tyr-Glu-Thr-Thr-Leu-Glu-Lys-Cys-Cys-Ala-Ala-Ala-Asp-Pro-His-Glu-Cys-Tyr-Ala- Lys-Val-Phe-Asp-Glu-Phe-Lys-Pro-Leu-Val-Glu-Glu-Pro-GIn-Asn-Leu-lle-Lys-GIn-Asn-Cys-Glu- Leu-Phe-Glu-GIn-Leu-Gly-Glu-Tyr-Lys-Phe-GIn-Asn-Ala-Leu-Leu-Val-Arg-Tyr-Thr-Lys-Lys-Val- Pro-GIn-Val-Ser-Thr-Pro-Thr-Leu-Val-Glu-Val-Ser-Arg-Asn-Leu-Gly-Lys-Val-Gly-Ser-Lys-Cys- Cys-Lys-His-Pro-Glu-Ala-Lys-Arg-Met-Pro-Cys-Ala-Glu-Asp-Tyr-Leu-Ser-Val-Val-Leu-Asn-GIn- Leu-Cys-Val-Leu-His-Glu-Lys-Thr-Pro-Val-Ser-Asp-Arg-Val-Thr-Lys-Cys-Cys-Thr-Glu-Ser-Leu- Val-Asn-Arg-Arg-Pro-Cys-Phe-Ser-Ala-Leu-Glu-Val-Asp-Glu-Thr-Tyr-Val-Pro-Lys-Glu-Phe-Asn- Ala-Glu-Thr-Phe-Thr-Phe-His-Ala-Asp-lle-Cys-Thr-Leu-Ser-Glu-Lys-Glu-Arg-GIn-lle-Lys-Lys- Gln-Thr-Ala-Leu-Val-Glu-Leu-Val-Lys-His-Lys-Pro-Lys-Ala-Thr-Lys-Glu-GIn-Leu-Lys-Ala-Val- Met-Asp-Asp-Phe-Ala-Ala-Phe-Val-Glu-Lys-Cys-Cys-Lys-Ala-Asp-Asp-Lys-Glu-Thr-Cys-Phe- Ala-Glu-Glu-Gly-Lys-Lys-Leu-Val-Ala-Ala-Ser-GIn-Ala-Ala-Leu-Gly-LeuMBHB Ref. No.: 24-0691 -WO(disulfide bridges linking Cys113-Cys122, Cys135-Cys151, Cys150-Cys161, Cys184-Cys229, Cys228-Cys237, Cys260-Cys306, Cys305-Cys313, Cys325-Cys339, Cys338-Cys349, Cys376-Cys421, Cys420-Cys429, Cys452-Cys498, Cys497-Cys508, Cys521-Cys537, Cys536-Cys547, Cys574-Cys619, Cys618-Cys627)Amylin and analogs

[0083] Analogs to amylin, also known as islet amyloid polypeptide (IAPP), have also been developed for use as anti-obesity and anti-diabetes drugs. These analogs are similar to portions of the native amylin peptide but contain minor sequence variations and residue modifications. Such analogs include pramlintide and cagrilintide (AM833). These molecules exert agonist activity on amylin receptors and thereby activate those receptors. The sequence of the native amylin peptide can be found below.

[0084] Amylin (SEQ ID NO: 21)Lys-Cys-Asn-Thr-Ala-Thr-Cys-Ala-Thr-GIn-Arg-Leu-Ala-Asn-Phe-Leu-Val-His-Ser-Ser-Asn-Asn-Phe-Gly-Ala-lle-Leu-Ser-Ser-Thr-Asn-Val-Gly-Ser-Asn-Thr-Tyr-NH2 (disulfide bridge at Cys2-Cys7).

[0085] The sequence of cagrilintide (AM833) can be found below.

[0086] Cagrilintide (AM833) (SEQ ID NO: 22)[Eicosanedioic acid-Y-Glu]-Lys-Cys-Asn-Thr-Ala-Thr-Cys-Ala-Thr-Gln-Arg-Leu-Ala-Glu-Phe-Leu-Arg-His-Ser-Ser-Asn-Asn-Phe-Gly-Pro-lle-Leu-Pro-Pro-Thr-Asn-Val-Gly-Ser-Asn-Thr-Pro-NH2 (disulfide bridge at Cys3-Cys8)Dual GLP-1 and Glucagon Receptor (GCCR) agonists

[0087] Molecules that bind and exert agonist activity on both GLP-1 receptors and glucagon receptors (GCCR) have also been developed for use as anti-obesity and anti-diabetes drugs. Such molecules include cotadutide (MEDI0382) and modified oxyntomodulin peptides. The sequences of cotadutide (MEDI0382) and an exemplary OXM (oxyntomodulin analog) can be found below.

[0088] MEDI0382 (Cotadutide) (SEQ ID NO: 23)MBHB Ref. No.: 24-0691 -WOHis-Ser-Gln-Gly-Thr-Phe-Thr-Ser-Asp-[£-(palmitoyl-Y-Glu)-Lys]-Ser-Glu-Tyr-Leu-Asp-Ser-Glu- Arg-Ala-Arg-Asp-Phe-Val-Ala-Trp-Leu-Glu-Ala-Gly-Gly, wherein -[e-(palmitoyl-y-Glu)-Lys]- has the following structure:

[0089] The sequence of OXM (oxyntomodulin analog) can be found below.

[0090] OXM (oxyntomodulin analog) (SEQ ID NO: 24)His-Aib-GIn-Gly-Thr-Phe-Thr-Ser-Asp-Tyr-Ser-Lys-Tyr-Leu-Asp-Glu-Arg-Arg-Ala-Arg-Asp-Phe-Val-Glu-Trp-Leu-Leu-Asn-Thr-[£-BrAc-PEGi2-Lys]-NH2Apelin Analogs

[0091] Analogs to apelin have also been developed for use as anti-obesity and anti-diabetes drugs. These analogs are similar to portions of the native apelin peptide but contain minor sequence variations and residue modifications. Such analogs can include MM07 (Brame et al., 2015, Hypertension 65(4): 834-840). Other non-peptide apelin receptor agonists include ML233 (Shibagaki et al., 2020, Neurochem. Res. 45: 752-759). These molecules exert agonist activity on apelin receptors.

[0092] The sequence of MM07 can be found below.

[0093] MM07 (SEQ ID NO: 25)Cys-Arg-Pro-Arg-Leu-Cys-His-Lys-Gly-Pro-Met-Pro-Phe(disulfide bridge, Cys1-Cys6)

[0094] Therapeutic molecules in this space are often therapeutic peptides, but need not be. They can be small molecules or antibodies.MBHB Ref. No.: 24-0691 -WONovel GLP-1 Analogs Containing Non-Proteinogenic Residues

[0095] Novel peptide analogs of GLP-1 are disclosed herein. The sequences and structures of those analogs are disclosed below. Each of these analogs can contain at least one non-proteinogenic amino acid, some of which are linked to heterologous moieties or substituents.

[0096] In one embodiment, the isolated peptide disclosed herein comprises the amino acid sequence:Xaa1-Xaa2-Xaa3-Xaa4-Xaa5-Xaa6-Xaa7-Xaa8-Xaa9-Xaa10-Xaa11-Xaa12-Xaa13-Xaa14-Xaa15-Xaa16-Xaa17-Xaa18-Xaa19-Xaa20-Xaa21 -Xaa22-Xaa23-Xaa24-Xaa25-Xaa26-Xaa27-Xaa28-Xaa29-Xaa30-Xaa31-Xaa32-Xaa33-Xaa34-Xaa35-Xaa36-Xaa37-Xaa38-Xaa39-Xaa40 (SEQ ID NO: 1); wherein: Xaa1 is His, 3-Mal-PEGo-2o-Lys, 3-Mal-Lys, Tyr, pyroGlu, or 3-Mal-PEGo-20; Xaa2 is Aib, Gly, Cys, Ser, Arg, or hArg; Xaa3 is Glu, Asn, Gin, Pro, or D-Arg; Xaa4 is Gly, Thr, Arg, or Gin; Xaa5 is Thr, Ala, Leu, or hArg; Xaa6 is Phe, Thr, Ser, or Pro; Xaa7 is Thr, Cys, His, or D-Arg; Xaa8 is Ser, Ala, £-3-Mal-PEGo-2o-Lys, or NMe-Leu; Xaa9 is Asp, Thr, Gly, or Ser; Xaa10 is Vai, Glu, Tyr, £-3-Mal-PEGo-2o-Lys, Pro, or His; Xaa11 is Ser, Arg, Met, or Lys; Xaa12 is Ser, Leu, lie, Glu, Lys, Pro, or Gly; Xaa13 is Tyr, Ala, Aib, Phe, or Oic; Xaa14 is Leu, Glu, NH, or pl-Phe; Xaa15 is Glu, Phe, Asp, Pro, or is absent; Xaa16 is Aib, Gly, Glu, Leu, Lys, Ser, D-Bip, or is absent; Xaa17 is Gin, Arg, lie, Glu, OH, or is absent; Xaa18 is Ala, His, Arg, or is absent; Xaa19 is Ala, Ser, Gin, or is absent; Xaa20 is Lys, Ser, £-3-Mal-PEGo-2o-Lys, Arg, or is absent; Xaa21 is Glu, Asn, Ala, Asp, or is absent; Xaa22 is Phe, Asn, or is absent; Xaa23 is lie, Phe, Vai, or is absent; Xaa24 is Ala, Gly, Gin, Ala, Glu, or is absent; Xaa25 is Trp, Pro, Thr, or is absent; Xaa26 is Leu, lie, or is absent; Xaa27 is Vai, Leu, lie, Glu, or is absent; Xaa28 is Lys, Pro, Ala, Asn, or is absent; Xaa29 is Gly, Thr, Pro, or is absent; Xaa30 is Gly, Arg, Asn, Thr, £-BrAc-PEGo-20-Lys, PEGo-2o-[£-3-Mal-D-Lys], oris absent; Xaa31 is Gly, (Sar)2.20-[£-3-Mal-Lys], Vai, Asn, Pro, OH, NH2, or is absent; Xaa32 is (Gly-Gly-Gly-Gly-Ser)3-[£-BrAc-Lys], PEGo-2o-(£-3-Mal-Lys), NH2, (Sar)2.20-[£-3-Mal-Lys], Gly, Vai, Ser, or is absent; Xaa33 is NH2, Ser, Gly, or is absent; Xaa34 is Asn, Ser, Gly, or is absent; Xaa35 is Thr, Asn, Ala, or is absent; Xaa36 is Pro, Thr, or is absent; Xaa37 is NH2, Pro, or is absent; Xaa38 is NH2, Pro, or is absent; Xaa39 is Ser or is absent; and Xaa40 is NH2or is absent.

[0097] In one aspect, the peptide disclosed herein is SBI-401, wherein Xaa1 is His, Xaa2 is Aib, Xaa3 is Glu, Xaa4 is Gly, Xaa5 is Thr, Xaa6 is Phe, Xaa7 is Thr, Xaa8 is Ser, Xaa9 is Asp, Xaa10 is Vai, Xaa11 is Ser, Xaa12 is Ser, Xaa13 is Tyr, Xaa14 is Leu, Xaa15 is Glu, Xaa16 is Aib, Xaa17 is Gin, Xaa18 is Ala, Xaa19 is Ala, Xaa20 is Lys, Xaa21 is Glu, Xaa22 is Phe, Xaa23 is lie, Xaa24 is Ala, Xaa25 is Trp, Xaa26 is Leu, Xaa27 is Vai, Xaa28 is Lys, Xaa29 is Gly, Xaa30MBHB Ref. No.: 24-0691 -WOis Gly, Xaa31 is Gly, Xaa32 is (Gly-Gly-Gly-Gly-Ser)3-[£-BrAc-Lys], Xaa33 is NH2, and Xaa34-Xaa40 are absent relative to SEQ ID NO: 1.

[0098] The amino acid sequence for SBI-401 (SEQ ID NO: 2) is shown below.His-Aib-Glu-Gly-Thr-Phe-Thr-Ser-Asp-Val-Ser-Ser-Tyr-Leu-Glu-Aib-GIn-Ala-Ala-Lys-Glu-Phe- lle-Ala-Trp-Leu-Val-Lys-Gly-Gly-Gly-(Gly-Gly-Gly-Gly-Ser)3-[£-BrAc-Lys]-NH2, wherein -[£-BrAc- Lys]-NH2has the following structure:

[0099] In another aspect, the peptide disclosed herein is SBI-402, wherein Xaa1 is His, Xaa2 is Aib, Xaa3 is Glu, Xaa4 is Gly, Xaa5 is Thr, Xaa6 is Phe, Xaa7 is Thr, Xaa8 is Ser, Xaa9 is Asp, Xaa10 is Vai, Xaa11 is Ser, Xaa12 is Ser, Xaa13 is Tyr, Xaa14 is Leu, Xaa15 is Glu, Xaa16 is Aib, Xaa17 is Gin, Xaa18 is Ala, Xaa19 is Ala, Xaa20 is Lys, Xaa21 is Glu, Xaa22 is Phe, Xaa23 is lie, Xaa24 is Ala, Xaa25 is Trp, Xaa26 is Leu, Xaa27 is Vai, Xaa28 is Lys, Xaa29 is Gly, Xaa30 is Gly, Xaa31 is Gly, Xaa32 is PEGi6-(£-3-Mal-Lys), Xaa33 is NH2, and Xaa34-Xaa40 are absent relative to SEQ ID NO: 1.

[0100] The amino acid sequence for SBI-402 (SEQ ID NO: 3) is shown below.

[0101] His-Aib-Glu-Gly-Thr-Phe-Thr-Ser-Asp-Val-Ser-Ser-Tyr-Leu-Glu-Aib-GIn-Ala-Ala-Lys-Glu-Phe-lle-Ala-Trp-Leu-Val-Lys-Gly-Gly-Gly-[PEGi6-(£-3-Mal-Lys)]-NH2, wherein -[PEGI6-(£-3-Mal-Lys)]-NH2has the following structure:MBHB Ref. No.: 24-0691 -WO

[0102] In another aspect, the peptide disclosed herein is SBI-403, wherein Xaa1 is His, Xaa2 is Gly, Xaa3 is Glu, Xaa4 is Gly, Xaa5 is Thr, Xaa6 is Phe, Xaa7 is Thr, Xaa8 is Ser, Xaa9 is Asp, Xaa10 is Vai, Xaa11 is Ser, Xaa12 is Ser, Xaa13 is Tyr, Xaa14 is Leu, Xaa15 is Glu, Xaa16 is Gly, Xaa17 is Gin, Xaa18 is Ala, Xaa19 is Ala, Xaa20 is Lys, Xaa21 is Glu, Xaa22 is Phe, Xaa23 is lie, Xaa24 is Ala, Xaa25 is Trp, Xaa26 is Leu, Xaa27 is Vai, Xaa28 is Lys, Xaa29 is Gly, Xaa30 is Arg, Xaa31 is (Sar)i6-[£-3-Mal-Lys], Xaa32 is NH2, and Xaa33-Xaa40 are absent relative to SEQ ID NO: 1.

[0103] The amino acid sequence for SBI-403 (SEQ ID NO: 4) is shown below.His-Gly-Glu-Gly-Thr-Phe-Thr-Ser-Asp-Val-Ser-Ser-Tyr-Leu-Glu-Gly-GIn-Ala-Ala-Lys-Glu-Phe- lle-Ala-Trp-Leu-Val-Lys-Gly-Arg-(Sar)i6-[£-3-Mal-Lys]-NH2, wherein -[£-3-Mal-Lys]-NH2 has the following structure:

[0104] In another aspect, the peptide disclosed herein is SBI-404, wherein Xaa1 is His, Xaa2 is Gly, Xaa3 is Glu, Xaa4 is Gly, Xaa5 is Thr, Xaa6 is Phe, Xaa7 is Thr, Xaa8 is Ser, Xaa9 is Asp, Xaa10 is Vai, Xaa11 is Ser, Xaa12 is Ser, Xaa13 is Tyr, Xaa14 is Leu, Xaa15 is Glu, Xaa16 is Glu, Xaa17 is Gin, Xaa18 is Ala, Xaa19 is Ala, Xaa20 is Lys, Xaa21 is Glu, Xaa22 is Phe, Xaa23 is lie, Xaa24 is Ala, Xaa25 is Trp, Xaa26 is Leu, Xaa27 is Vai, Xaa28 is Lys, Xaa29 is Gly, Xaa30 is Gly, Xaa31 is Gly, Xaa32 is (Sar)i6-[£-3-Mal-Lys], Xaa33 is NH2, and Xaa34-Xaa40 are absent relative to SEQ ID NO: 1.

[0105] The amino acid sequence for SBI-404 (SEQ ID NO: 5) is shown below.His-Gly-Glu-Gly-Thr-Phe-Thr-Ser-Asp-Val-Ser-Ser-Tyr-Leu-Glu-Glu-GIn-Ala-Ala-Lys-Glu-Phe- lle-Ala-Trp-Leu-Val-Lys-Gly-Gly-Gly-(Sar)i6-[£-3-Mal-Lys]-NH2, wherein -[£-3-Mal-Lys]-NH2 has the following structure:MBHB Ref. No.: 24-0691 -WO

[0106] In another aspect, the peptide disclosed herein is SBI-405, wherein Xaa1 is 3-Mal- PEGs-Lys, Xaa2 is Cys, Xaa3 is Asn, Xaa4 is Thr, Xaa5 is Ala, Xaa6 is Thr, Xaa7 is Cys, Xaa8 is Ala, Xaa9 is Thr, Xaa10 is Gin, Xaa11 is Arg, Xaa12 is Leu, Xaa13 is Ala, Xaa14 is Glu, Xaa15 is Phe, Xaa16 is Leu, Xaa17 is Arg, Xaa18 is His, Xaa19 is Ser, Xaa20 is Ser, Xaa21 is Asn, Xaa22 is Asn, Xaa23 is Phe, Xaa24 is Gly, Xaa25 is Pro, Xaa26 is lie, Xaa27 is Leu, Xaa28 is Pro, Xaa29 is Pro, Xaa30 is Thr, Xaa31 is Asn, Xaa32 is Vai, Xaa33 is Gly, Xaa34 is Ser, Xaa35 is Asn, Xaa36 is Thr, Xaa37 is Pro, Xaa38 is NH2, Xaa39-Xaa40 are absent, wherein the peptide further contains a disulfide bridge, Cys2-Cys7 relative to SEQ ID NO: 1.

[0107] The amino acid sequence for SBI-405 (SEQ ID NO: 6) is shown below.[3-Mal-PEG8-Lys]-Cys-Asn-Thr-Ala-Thr-Cys-Ala-Thr-Gln-Arg-Leu-Ala-Glu-Phe-Leu-Arg-His-Ser-Ser-Asn-Asn-Phe-Gly-Pro-lle-Leu-Pro-Pro-Thr-Asn-Val-Gly-Ser-Asn-Thr-Pro-NH2 (disulfide bridge, Cys2-Cys7), wherein [3-Mal-PEGs-Lys] has the following structure:>

[0108] In another aspect, the peptide disclosed herein is SBI-406, wherein Xaa1 is 6-Mal-Lys, Xaa2 is Cys, Xaa3 is Asn, Xaa4 is Thr, Xaa5 is Ala, Xaa6 is Thr, Xaa7 is Cys, Xaa8 is Ala, Xaa9 is Thr, Xaa10 is Gin, Xaa11 is Arg, Xaa12 is Leu, Xaa13 is Ala, Xaa14 is Glu, Xaa15 is Phe, Xaa16 is Leu, Xaa17 is Arg, Xaa18 is His, Xaa19 is Ser, Xaa20 is Ser, Xaa21 is Asn, Xaa22 is Asn, Xaa23 is Phe, Xaa24 is Gly, Xaa25 is Pro, Xaa26 is lie, Xaa27 is Leu, Xaa28 is Pro, Xaa29 is Pro, Xaa30 is Thr, Xaa31 is Asn, Xaa32 is Vai, Xaa33 is Gly, Xaa34 is Ser, Xaa35 is Asn, Xaa36 is Thr, Xaa37 is Pro, Xaa38 is NH2, Xaa39-Xaa40 are absent, wherein the peptide further contains a disulfide bridge, Cys2-Cys7 relative to SEQ ID NO: 1.MBHB Ref. No.: 24-0691 -WO

[0109] The amino acid sequence for SBI-406 (SEQ ID NO: 7) is shown below.[6-Mal-Lys]-Cys-Asn-Thr-Ala-Thr-Cys-Ala-Thr-Gln-Arg-Leu-Ala-Glu-Phe-Leu-Arg-His-Ser-Ser-Asn-Asn-Phe-Gly-Pro-lle-Leu-Pro-Pro-Thr-Asn-Val-Gly-Ser-Asn-Thr-Pro-NH? (disulfide bridge, Cys2-Cys7), wherein [6-Mal-Lys]- has the following structure:

[0110] In another aspect, the peptide disclosed herein is SBI-407, wherein Xaa1 is Tyr, Xaa2 is Aib, Xaa3 is Glu, Xaa4 is Gly, Xaa5 is Thr, Xaa6 is Phe, Xaa7 is Thr, Xaa8 is Ser, Xaa9 is Asp, Xaa10 is Tyr, Xaa11 is Ser, Xaa12 is lie, Xaa13 is Aib, Xaa14 is Leu, Xaa15 is Asp, Xaa16 is Lys, Xaa17 is lie, Xaa18 is Ala, Xaa19 is Gin, Xaa20 is e-3-Mal-PEG8-Lys, Xaa21 is Ala, Xaa22 is Phe, Xaa23 is Vai, Xaa24 is Gin, Xaa25 is Trp, Xaa26 is Leu, Xaa27 is lie, Xaa28 is Ala, Xaa29 is Gly, Xaa30 is Gly, Xaa31 is Pro, Xaa32 is Ser, Xaa33 is Ser, Xaa34 is Gly, Xaa35 is Ala, Xaa36 is Pro, Xaa37 is Pro, Xaa38 is Pro, Xaa39 is Ser, Xaa40 is NH2 relative to SEQ ID NO: 1.

[0111] The amino acid sequence for SBI-407 (SEQ ID NO: 8) is shown below.Tyr-Aib-Glu-Gly-Thr-Phe-Thr-Ser-Asp-Tyr-Ser-lle-Aib-Leu-Asp-Lys-lle-Ala-Gln-[£-3-Mal-PEG8- Lys]-Ala-Phe-Val-Gln-Trp-Leu-lle-Ala-Gly-Gly-Pro-Ser-Ser-Gly-Ala-Pro-Pro-Pro-Ser-NH2, wherein -[E-3-Mal-PEG8-Lys] has the following structure:

[0112] In another aspect, the peptide disclosed herein is SBI-408, wherein Xaa1 is His, Xaa2 is Ser, Xaa3 is Gin, Xaa4 is Gly, Xaa5 is Thr, Xaa6 is Phe, Xaa7 is Thr, Xaa8 is Ser, Xaa9 is Asp, Xaa10 is £-3-Mal-PEG8-Lys, Xaa11 is Ser, Xaa12 is Glu, Xaa13 is Tyr, Xaa14 is Leu, Xaa15 is Asp, Xaa16 is Ser, Xaa17 is Glu, Xaa18 is Arg, Xaa19 is Ala, Xaa20 is Arg, Xaa21 is Asp, Xaa22MBHB Ref. No.: 24-0691 -WOis Phe, Xaa23 is Vai, Xaa24 is Ala, Xaa25 is T rp, Xaa26 is Leu, Xaa27 is Glu, Xaa28 is Ala, Xaa29 is Gly, Xaa30 is Gly, Xaa31 is OH, and Xaa32-Xaa40 are absent relative to SEQ ID NO: 1.

[0113] The amino acid sequence for SBI-408 (SEQ ID NO: 9) is shown below.His-Ser-Gln-Gly-Thr-Phe-Thr-Ser-Asp-[£-3-Mal-PEGs-Lys]-Ser-Glu-Tyr-Leu-Asp-Ser-Glu-Arg- Ala-Arg-Asp-Phe-Val-Ala-Trp-Leu-Glu-Ala-Gly-Gly-OH, wherein -[E-3-Mal-PEGs-Lys] has the following structure:

[0114] In another aspect, the peptide disclosed herein is SBI-409, wherein Xaa1 is His, Xaa2 is Aib, Xaa3 is Gin, Xaa4 is Gly, Xaa5 is Thr, Xaa6 is Phe, Xaa7 is Thr, Xaa8 is Ser, Xaa9 is Asp, Xaa10 is Tyr, Xaa11 is Ser, Xaa12 is Lys, Xaa13 is Tyr, Xaa14 is Leu, Xaa15 is Asp, Xaa16 is Glu, Xaa17 is Arg, Xaa18 is Arg, Xaa19 is Ala, Xaa20 is Arg, Xaa21 is Asp, Xaa22 is Phe, Xaa23 is Vai, Xaa24 is Glu, Xaa25 is Trp, Xaa26 is Leu, Xaa27 is Leu, Xaa28 is Asn, Xaa29 is Thr, Xaa30 is £-BrAc-PEGi2-Lys, Xaa31 is NH2, and Xaa32-Xaa40 are absent relative to SEQ ID NO: 1.

[0115] The amino acid sequence for SBI-409 (SEQ ID NO: 10) is shown below.His-Aib-GIn-Gly-Thr-Phe-Thr-Ser-Asp-Tyr-Ser-Lys-Tyr-Leu-Asp-Glu-Arg-Arg-Ala-Arg-Asp-Phe- Val-Glu-Trp-Leu-Leu-Asn-Thr-[£-BrAc-PEGi2-Lys]-NH2, wherein [£-BrAc-PEGi2-Lys]-NH2has the following structure:

[0116] In another aspect, the peptide disclosed herein is SBI-410, wherein Xaa1 is His, Xaa2 is Aib, Xaa3 is Gin, Xaa4 is Gly, Xaa5 is Thr, Xaa6 is Phe, Xaa7 is Thr, Xaa8 is Ser, Xaa9 is Asp, Xaa10 is Tyr, Xaa11 is Ser, Xaa12 is Lys, Xaa13 is Tyr, Xaa14 is Leu, Xaa15 is Asp, Xaa16 isMBHB Ref. No.: 24-0691 -WOGlu, Xaa17 is Arg, Xaa18 is Arg, Xaa19 is Ala, Xaa20 is Arg, Xaa21 is Asp, Xaa22 is Phe, Xaa23 is Vai, Xaa24 is Glu, Xaa25 is Trp, Xaa26 is Leu, Xaa27 is Leu, Xaa28 is Asn, Xaa29 is Thr, Xaa30 is PEGs-[£-3-Mal-D-Lys], Xaa31 is NH2, and Xaa32-Xaa40 are absent relative to SEQ ID NO: 1.

[0117] The amino acid sequence for SBI-410 (SEQ ID NO: 11) is shown below.His-Aib-GIn-Gly-Thr-Phe-Thr-Ser-Asp-Tyr-Ser-Lys-Tyr-Leu-Asp-Glu-Arg-Arg-Ala-Arg-Asp-Phe-Val-Glu-Trp-Leu-Leu-Asn-Thr-PEG8-[£-3-Mal-D-Lys]-NH2, wherein -[PEG8-£-3-Mal-D-Lys]-NH2has the following structure:10118] In another aspect, the peptide disclosed herein is SBI-411, wherein Xaa1 is pyroGlu, Xaa2 is Arg, Xaa3 is Pro, Xaa4 is Arg, Xaa5 is Leu, Xaa6 is Ser, Xaa7 is His, Xaa8 is c-3-Mal- PEGs-Lys, Xaa9 is Gly, Xaa10 is Pro, Xaa11 is Met, Xaa12 is Pro, Xaa13 is Phe, Xaa14 is NH2, and Xaa15-Xaa40 are absent relative to SEQ ID NO: 1.

[0119] The amino acid sequence for SBI-411 (SEQ ID NO: 12) is shown below.[pyroGlu]-Arg-Pro-Arg-Leu-Ser-His-[£-3-Mal-PEG8-Lys]-Gly-Pro-Met-Pro-Phe-NH2, wherein -[£-3-Mal-PEGs-Lys] has the following structure:

[0120] In another aspect, the peptide disclosed herein is SBI-412, wherein Xaa1 is 3-Mal- PEG8, Xaa2 is hArg, Xaa3 is D-Arg, Xaa4 is Gin, Xaa5 is hArg, Xaa6 is Pro, Xaa7 is D-Arg, Xaa8 is NMe-Leu, Xaa9 is Ser, Xaa10 is His, Xaa11 is Lys, Xaa12 is Gly, Xaa13 is Oic, Xaa14 is pl-MBHB Ref. No.: 24-0691 -WOPhe, Xaa15 is Pro, Xaa16 is D-Bip, Xaa17 is OH, and Xaa18-Xaa40 are absent relative to SEQ ID NO: 1.

[0121] The amino acid sequence for SBI-412 (SEQ ID NO: 13) is shown below.[3-Mal-PEG8]-hArg-[D-Arg]-Gln-hArg-Pro-[D-Arg]-(A / Me-Leu]-Ser-His-Lys-Gly-Oic-[pl-Phe]-Pro-[D-Bip]-OH, wherein [3-Mal-PEGs] has the following structure shown above, wherein the complete structural formula of SBI-412 is as follows:

[0122] In another embodiment, at least one of the peptides SBI-401 (SEQ ID NO: 2), SBI-402 (SEQ ID NO: 3), SBI-403 (SEQ ID NO: 4), SBI-404 (SEQ ID NO: 5), SBI-405 (SEQ ID NO: 6), SBI-406 (SEQ ID NO: 7), SBI-407 (SEQ ID NO: 8), SBI-408 (SEQ ID NO: 9), SBI-409 (SEQ ID NO: 10), SBI-410 (SEQ ID NO: 11), SBI-411 (SEQ ID NO: 12), and SBI-412 (SEQ ID NO: 13) is used in a pharmaceutical composition, further comprising a pharmaceutically acceptable excipient, carrier, or diluent.

[0123] In another embodiment, at least one of the peptides SBI-401 (SEQ ID NO: 2), SBI-402 (SEQ ID NO: 3), SBI-403 (SEQ ID NO: 4), SBI-404 (SEQ ID NO: 5), SBI-405 (SEQ ID NO: 6), SBI-406 (SEQ ID NO: 7), SBI-407 (SEQ ID NO: 8), SBI-408 (SEQ ID NO: 9), SBI-409 (SEQ ID NO: 10), SBI-410 (SEQ ID NO: 11), SBI-411 (SEQ ID NO: 12), and SBI-412 (SEQ ID NO: 13) or the pharmaceutical composition comprising at least one of the peptides SBI-401 (SEQ ID NO: 2), SBI-402 (SEQ ID NO: 3), SBI-403 (SEQ ID NO: 4), SBI-404 (SEQ ID NO: 5), SBI-405 (SEQ ID NO: 6), SBI-406 (SEQ ID NO: 7), SBI-407 (SEQ ID NO: 8), SBI-408 (SEQ ID NO: 9), SBI-409MBHB Ref. No.: 24-0691 -WO(SEQ ID NO: 10), SBI-410 (SEQ ID NO: 11), SBI-411 (SEQ ID NO: 12), and SBI-412 (SEQ ID NO: 13), and further comprising a pharmaceutically acceptable excipient, carrier, or diluent is used in a kit.

[0124] In various embodiments, the kit further comprises buffers, reducing agents, activation reagents, catalysts, enzymes, or other auxiliary components that facilitate conjugation, purification, or analysis of the resulting ADCs. Such auxiliary components may include, for example, phosphate-buffered saline, acetate buffers, reducing agents (e.g., TCEP), stabilizers, or quenching reagents.

[0125] In various embodiments, the kit components may be packaged separately or in combination, optionally in sterile vials, ampoules, or other suitable containers. Each component may be packaged in an amount that, when combined according to the instructions provided, yields reaction conditions sufficient to obtain an ADC with a predetermined DAR or DAR distribution.

[0126] In various embodiments, the kit may be configured to enable site-specific conjugation or controlled conjugation (e.g., via engineered cysteine residues, enzymatic ligation, or clickchemistry-based linkers). Such kits may further include reagents for site activation, selective reduction, or enzymatic processing.

[0127] The disclosure further encompasses kits comprising analytical reagents, standards, or reference materials suitable for determining DAR values or verifying DAR distributions of ADCs produced according to the instructions provided.Antibody-Drug Conjugates

[0128] Antibody-drug conjugates (ADCs) are an emerging therapeutic modality with applications in a wide variety of disease areas. Antibody-drug conjugates are also known as immunoconjugates. These drug constructs comprise an antibody, or antigen-binding fragment thereof, with specificity to a particular disease-related antigen, allowing for localization of the construct to only cells relevant for the pathogenesis of a disease. The antibody or antibody fragment is linked to an active drug molecule that is delivered to the local cell environment upon binding of the antibody to its target. The active drug molecule can be a small molecule, an oligonucleotide, a peptide, or the like, and linking the active drug molecule to the antibody allows for precise delivery of the drug to relevant cells without circulating the drug throughout the body, often at concentrations much lower than drugs that are delivered systemically. The drug molecule can be a cytotoxic agent or some other molecule with a therapeutic effect. The use of ADCs hasMBHB Ref. No.: 24-0691 -WOseveral advantages, including (1) alleviating off-target effects in non-disease related tissues and (2) reducing dose-dependent on-target effects by allowing for lower therapeutic doses.Drug-to-Antibody Ratios (DAR)

[0129] A commonly measured parameter related to ADC pharmacology and efficacy is the drug-to-antibody ratio or drug-antibody ratio (DAR). When discussing antibody-drug conjugates, this parameter can also be known as the peptide-to-antibody ratio. This ratio represents the average number of drug (or peptide) molecules covalently conjugated per antibody molecule within a preparation of antibody-drug conjugates (ADCs). In some embodiments, the DAR is increased by conjugating multiple copies of a peptide to each other. The DAR value can vary between batches of manufactured ADC but can be approximately controlled through various antibody engineering and conjugation chemistry methods. The DAR typically varies between about 0 and about 8, inclusive. The DAR for an ADC can be about 1, about 2, about 3, about 4, about 5, about 6, about 7, about 8, about 9, or about 10. In some embodiments, the DAR is about 2 or about 4. In some embodiments, the DAR is less than 1 , such as about 0.8, about 0.6, about 0.4, about 0.2, or about 0.1. Methods of determining DAR and reducing variability thereof are discussed in U.S. Patent Publication 2023 / 0364252 A1, incorporated by reference herein, among others, and are well known in the art.

[0130] Also provided herein are methods of producing ADCs with defined DAR values or DAR distributions. Such methods may include adjusting one or more conjugation parameters, including but not limited to: (i) the molar equivalents of drug-linker reagent relative to the antibody; (ii) the extent of antibody reduction (e.g., partial vs. full interchain disulfide reduction); (iii) the identity and concentration of reducing agents; (iv) reaction temperature; (v) reaction pH; (vi) reaction time; and / or (vii) enzymatic activation or site-specific conjugation conditions. These parameters may be modulated individually or in combination to yield ADCs with targeted DARs, reduced DAR heterogeneity, or improved lot-to-lot consistency.

[0131] In some embodiments, the methods described herein produce ADCs having an average DAR of about 1-4, about 2-6, about 3-8, or about 4-10. In some embodiments, the DAR is less than 1, such as about 0.8, about 0.6, about 0.4, about 0.2, or about 0.1. In further embodiments, the methods yield ADC compositions enriched for a desired DAR species, such as DAR 2 or DAR 4. The methods may be used to reproducibly obtain ADCs with predetermined DAR specifications suitable for therapeutic, diagnostic, or research use.MBHB Ref. No.: 24-0691 -WOCysteine-based mAb bioconjugation

[0132] Bioconjugation through cysteine sulfhydryl groups is an effective strategy that does not require expensive and time-consuming reengineering of the antibody structure. In cysteine-based conjugation, the antibody is first treated with a reducing agent, such as dithiothreitol (DTT) or tris (2-carboxyethyl) phosphine (TCEP), which can convert the disulfides of an antibody to free cysteine residues (Hamblett et al., 2004, Clin. Cancer Res. 10(20): 7063-7070). There are 16 cysteine pairs in a full IgG 1 or a lgG4 scaffold (Figure 1): 12 intra-chain (in red) and 4 inter-chain disulfide bonds. Due to greater solvent accessibility, the four inter-chain disulfide bonds are significantly more susceptible to reduction by reducing agents than are the intra-chain bonds (Willner et al., 1993, Bioconjugate Chem. 4(6): 521-527). These inter-chain cysteine residues are distant from the antigen-binding variable domains, and the modification with a payload usually does not impact antibody’s antigen binding.

[0133] After reduction, the sulfhydryl (SH) group of the antibody can be conjugated with maleimide-containing (FIG. 2) or alpha-bromoacetamido-containing (FIG. 3) payloads to form antibody-drug conjugates (ADCs) under biocompatible weak basic conditions.

[0134] An ADC with a drug-to-antibody ratio of 8 (DAR8) (all the 8 inter-chain sulfhydryl groups are linked to a payload) was shown to have less desirable pharmacokinetic and toxicologic properties due to its aggregation and short plasma elimination half-life [Hamblett2004, McDonagh2006], The partial reduction of 4 inter-chain disulfide bonds strategy is usually used. Because the reactivities of these 4 pairs of inter-chain disulfide bonds are similar, the free sulfhydryl groups are produced randomly under partial reduction conditions. Therefore, cysteine-based bioconjugation results in a heterogeneous mixture of conjugation products with varying sites and number of drugs attached [Hamblett2004, Sun2005] (FIG. 4, FIG. 5).

[0135] In addition to the methods described above, rational design and protein engineering can enable site-specific — i.e., non-random — drug conjugation, which allows for precise control of the DAR by conjugating molecules only at predetermined residues. This can allow for less variability and better efficacy of the final ADC product. In some embodiments, the drug is conjugated to the N-terminus of either the light chain variable region or the heavy chain variable region. Methods of accomplishing this are known to those in the art (see, for example, Hingorani, 2024, “An overview of site-specific methods for achieving antibody drug conjugates with homogenous drug to antibody ratio,” Expert Opin. Biol. Then 24: 31-36).MBHB Ref. No.: 24-0691 -WOOther mAb bioconjugation strategies

[0136] In some embodiments, the drug is conjugated at any nucleophilic site. Drug molecules can be covalently conjugated to antibodies at any site with a reactive nucleophile, including the N-terminus; amino groups on residue side chains, particularly the e-amino group of lysine residues; various non-proteinogenic residue side groups; and carboxyl groups. Other manners of conjugating antibodies to drug molecules are known to those in the art.Linkers

[0137] The antibody portion of an ADC and the drug portion of an ADC can be linked to each other. They can also be attached to each other, fused to each other, or conjugated to each other. The constituent portions of an ADC can be linked, attached, fused, or conjugated to each other by a moiety or molecule called a linker. Linkers are also known as spacers. The linker can be a peptide or a series of peptides. The peptide linker can be glycine- rich, which imparts flexibility to the conjugate and allows for a greater number of binding conformations. The peptide linker can be cleavable, such as by proteases, or non-cleavable. Cleavable linkers allow for controlled release of the drug into or around cells. Linkers can be non-immunogenic, and they can be covalently bonded to the antibody portion and / or the drug portion of the ADC. Peptide linkers can comprise natural / proteinogenic amino acids, unnatural / non-proteinogenic amino acids, or a mixture of natural / proteinogenic amino acids and unnatural / non-proteinogenic amino acids. Linkers can be hydrophilic linkers. The term “hydrophilic linker” as used herein means a spacer molecule comprising a chemical moiety which comprises at least 5 non-hydrogen atoms where 30-50% of these are either N or O. For examples, see U.S. Patent 8,129,343, incorporated by reference herein, among others. Other examples of linkers contain gamma glutamic acid (gamma glutamate), fatty acids and fatty acid esters, p-alanine, aminohexanoic acid, urethane, or cholesterol moieties, among others. Linkers can also comprise PEG (polyethylene glycol) moieties of various lengths.

[0138] In some embodiments, the number of non-proteinogenic residues in a linker is variable. Such residues may constitute at least a portion of a linker between the antibody and the peptide and may be selected to modulate linker length, flexibility, solubility, and / or pharmacokinetic properties. In certain embodiments, the linker comprises one or more sarcosine residues. The number of sarcosine residues may range from, for example but not limited to, 0 toMBHB Ref. No.: 24-0691 -WO20 residues. In some embodiments, the linker comprises 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 sarcosine residues. In certain embodiments, the linker comprises one or more polyethylene glycol (PEG) units. The number of PEG units may range from, for example but not limited to, 0 to 20 units,. In some embodiments, the PEG comprises 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 PEG units.Residue Derivatization and Substituents

[0139] Peptide residues as disclosed herein can be associated with a heterologous moiety, e.g., to extend in vivo half-life, or to impart some other favorable therapeutic property. Heterologous moieties can also be known as substituents. The heterologous moiety or substituent can be a protein, a peptide, a protein domain, a linker, an organic (carbon containing) polymer, an inorganic (non-carbon containing) polymer, a polyethylene glycol (PEG) moiety, biotin, albumin, human serum albumin (HSA), a HSA FcRn binding portion, an antibody, a domain of an antibody, an antibody fragment, a single chain antibody, a domain antibody, an albumin binding domain, an enzyme, a ligand, a receptor, a binding peptide, a non-Fnlll scaffold, an epitope tag, a recombinant polypeptide polymer, a cytokine, or a combination of two or more of such moieties. Amino acid or peptide residues which have been chemically modified through conjugation to one or more of the heterologous moieties or substituents described herein are known as derivatives, and the process of so modifying peptide residues is called derivatization.Anti-Cannabinoid Receptor 1 (CB1) Antibodies

[0140] In various aspects of the disclosure, antibodies for use in conjugates according to the disclosure bind to extra-cellular loop 2 (ECL2) of human CB1.

[0141] In various embodiments of the disclosure, the anti-CB1 antibodies include a heavy chain variable region (VH or VH region) comprising the following complementarity determining regions (CDRs): HCDR1, SEQ ID NO: 26; HCDR2, SEQ ID NO: 27; and HCDR3, SEQ ID NO: 28. In various embodiments of the disclosure, the anti-CB1 antibodies include a light chain variable region (VL or VL region) comprising the following complementarity determining regions (CDRs): LCDR1 , SEQ ID NO: 29; LCDR2, SEQ ID NO: 30; LCDR3, SEQ ID NO: 31. This set of light chain and heavy chain CDRs is designated herein as the P1C4 CDRs.MBHB Ref. No.: 24-0691 -WO

[0142] In some embodiments, the isolated antibody or fragment thereof comprises a heavy chain CDR1 sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% homology to the amino acid sequence of SEQ ID NO: 26 (YYWMN). In another embodiment, the isolated antibody orfragment thereof comprises a heavy chain CDR2 sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% homology to the amino acid sequence of SEQ ID NO: 27 (QIYPGDGETKY). In another embodiment, the isolated antibody or fragment thereof comprises a heavy chain CDR3 sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% homology to the amino acid sequence of SEQ ID NO: 28 (SHGNYLPY). In another embodiment, the isolated antibody or fragment thereof comprises a light chain CDR1 sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% homology to the amino acid sequence of SEQ ID NO: 29 (SSYLH). In another embodiment, the isolated antibody or fragment thereof comprises a light chain CDR2 sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% homology to the amino acid sequence of SEQ ID NO: 30 (STSNLAS). In another embodiment, the isolated antibody or fragment thereof comprises a light chain CDR3 sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% homology to the amino acid sequence of SEQ ID NO: 31 (HQYHRSPPTF).

[0143] In embodiments, one or more of the P1C4 CDRs may include an amino acid substitution as provided in Table 1 for heavy chain CDRs and Table 2 for light chain CDRs. Variant anti-CB1 antibodies having one of the mutant CDRs from Tables 1 and 2 bound to human CB1 with at least 50% of the binding affinity of the humanized P1C4 antibody. Non-exhaustive examples of modified anti-CB1 antibodies are disclosed in U.S. Application No. 14 / 774,582, filed September 10, 2015 (published as US20170210797A1), International Application No. PCT / US15 / 23108, filed March 27, 2015 (published as WO2015148984A2), International Application No. PCTUS2016 / 053927, filed September 26, 2016 (published as WQ2017058771A1), and US Application No. 15 / 765,135 filed October 8, 2018 (published as US20180282406A1), the disclosures of each of which are incorporated herein by reference in their entireties.Table 1P1C4 Heavy Chain CDRs and Single Amino Acid Substitution VariantsMBHB Ref. No.: 24-0691 -WO>>>>>>>>>>>>>>>MBHB Ref. No.: 24-0691 -WO>>>>>>>>>>>>>>>>>MBHB Ref. No.: 24-0691 -WO>>>>>>>>>>>>>>>>>MBHB Ref. No.: 24-0691 -WO>>>>>>>>>>>>>>>>>>>>>>>>>>>>>>MBHB Ref. No.: 24-0691 -WO>Table 2P1C4 Light Chain CDRs and Single Amino Acid Substitution Variants>>>>>>>>>>>>>>>>>>>>>>>>>> MBHB Ref. No.: 24-0691 -WO>>>>>>>>>>>>>>>>>>>>>>>>>>>>>>>>>>>>>> MBHB Ref. No.: 24-0691 -WO>>>>>>>>>>>>>>>>>>>>>>MBHB Ref. No.: 24-0691 -WO>>>>>>>MBHB Ref. No.: 24-0691 -WO>>>>>>>>>>>>>>>>>

[0144] In embodiments, the anti-CB1 antibodies for use according to the disclosure include the P1C4 CDRs (SEQ ID NOS 26-31) wherein one, two, or all three of the P1C4 heavy chain CDRs are replaced variant CDR(s) selected from Table 1 and / or one, two, or three lights chain CDRs are replaced with variant CDR(s) selected from Table 2.

[0145] In other embodiments of the anti-CB1 antibodies according to the disclosure include antibodies having CDRs identified by affinity maturation of the humanized P1C4 antibody as more fully described in US Patent Application Publication No. US20180282406A1 as shown in Table 2.Table 3Summary of Amino Acid Changes in Affinity Matured Clones>>>>> MBHB Ref. No.: 24-0691 -WO>

[0146] In still another embodiment, the isolated antibody or antigen binding fragment thereof comprises a light chain CDR1 comprising amino acid sequence according to SEQ ID NO: 238. In further embodiments, the isolated antibody or antigen binding fragment thereof comprises a light chain CDR2 selected from a group comprising amino acid sequence according to SEQ ID NOs: 255, 287, and 301. In still further embodiments, the isolated antibody or antigen binding fragment thereof comprises a light chain CDR3 selected from a group comprising amino acid sequence according to SEQ ID NOs: 346 and 374.

[0147] In an embodiment, the isolated antibody or antigen binding fragment thereof comprises a light chain CDR1 comprising amino acid sequence according to SEQ ID NO: 238, a light chain CDR2 comprising amino acid sequence according to SEQ ID NO: 287, and a light chain CDR3 comprising amino acid sequence according to SEQ ID NO: 346.

[0148] In another embodiment, the isolated antibody or antigen binding fragment thereof comprises a light chain CDR1 comprising amino acid sequence according to SEQ ID NO: 238, a light chain CDR2 comprising amino acid sequence according to SEQ ID NO: 301, and a light chain CDR3 comprising amino acid sequence according to SEQ ID NO: 346.

[0149] In yet another embodiment, the isolated antibody or antigen binding fragment thereof comprises a light chain CDR1 comprising amino acid sequence according to SEQ ID NO: 238, a light chain CDR2 comprising amino acid sequence according to SEQ ID NO: 287, and a light chain CDR3 comprising amino acid sequence according to SEQ ID NO: 31.

[0150] In still another embodiment, the isolated antibody or antigen binding fragment thereof comprises a light chain CDR1 comprising amino acid sequence according to SEQ ID NO: 238, a light chain CDR2 comprising amino acid sequence according to SEQ ID NO: 255, and a light chain CDR3 comprising amino acid sequence according to SEQ ID NO: 374.

[0151] In still another embodiment, disclosed herein is an isolated antibody or antigen binding fragment thereof that binds to cannabinoid receptor 1 (CB1), wherein the antibody or antigen binding fragment thereof comprises a light chain CDR1 comprising an amino acid sequence according to SEQ ID NOs: 29 or 238; a light chain CDR2 comprising an amino acid sequence according to SEQ ID NOs: 30, 255, 287, or 301; and a light chain CDR3 comprising an amino acid sequence according to SEQ ID NOs: 31, 346, or 374.MBHB Ref. No.: 24-0691 -WO

[0152] The person of skill in the art will understand that the heavy and light chain CDRs of the antibodies provided herein may be independently selected, or mixed and matched, to form an antibody or binding fragment thereof comprising any light chain CDR1, CDR2, and CDR3; and any heavy chain CDR1 , CDR2, and CDR3 from the antibodies provided herein. The skilled person will further understand that the heavy and light chain variable regions of the antibodies provided herein may be independently selected, or mixed and matched, to form an antibody or binding fragment comprising any heavy and light chain from the antibodies provided herein.

[0153] In some embodiments, the antibody or antigen binding fragment thereof provided herein is a chimeric antibody or fragment containing heavy and light chain CDRs selected from the CDRs provided herein, or conservative variants of the CDRs provided herein. In another embodiment, the antibody or antigen binding fragment thereof provided herein is a humanized antibody or fragment containing heavy and light chain CDRs selected from the CDRs provided herein, or conservative variants of the CDRs provided herein. In some embodiments, the antibody or antigen binding fragment thereof provided herein comprises a light chain and / or a heavy chain comprising a sequence provided herein, or a conservative variant thereof. In some embodiments, the conservative variants have at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% homology to the reference sequence provided herein. In some embodiments, the conservative variants comprise 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or more amino acid substitutions, insertions, or deletions.

[0154] Humanized anti-CB1 antibodies comprising the heavy and light chain CDRs provided herein, or variants thereof, may be generated using any human framework sequence, and are also encompassed in the present disclosure. In some embodiments, framework sequences suitable for use in the present disclosure include those framework sequences that are structurally similar to the framework sequences provided herein. In some embodiments, human frameworks are selected based on homology between the parent antibody and the human germline VH and VK genes. Selected frameworks, in some embodiments, have the highest homology with the parent antibody VH and VK genes and also are predicted, based on computer modeling or other means, to support the CDR structure predicted to be presented by the parent antibody.

[0155] Further modifications in the framework regions may be made to improve the properties of the antibodies provided herein. Such further framework modifications may include chemical modifications; point mutations to reduce immunogenicity or remove T cell epitopes; or back mutation to the residue in the original germline sequence. In some embodiments of the presentMBHB Ref. No.: 24-0691 -WOdisclosure, the humanized antibodies and fragments thereof comprise a human framework and grafted CDRs provided herein, without further modifications to the variable region.

[0156] The present disclosure also encompasses humanized antibodies that bind to CB1 and comprise framework modifications corresponding to the exemplary modifications described herein with respect to any suitable framework sequence, as well as other framework modifications that otherwise improve the properties of the antibodies. The CB1 antibodies and fragments thereof disclosed herein may be of an lgG1, lgG2, lgG3, or lgG4 isotype, or any combination thereof. The term “isotype” refers to the antibody class encoded by the heavy chain constant region genes. In addition, the heavy chain constant region may be derived from any species including, but not limited to, mouse, rat, rabbit, hamster, guinea pig, primate, llama or human. For example, in some embodiments, the CB1 antibodies and fragments thereof of the present disclosure comprise a human lgG1 Fc constant region. In another embodiment, the CB1 antibodies and fragments thereof comprise a human lgG2, human lgG4, or hybrid lgG2-lgG4 Fc constant region.

[0157] In some embodiments, theCBI antibody exhibits reduced effector function. In a further embodiment, the CB1 antibody comprises an lgG4 Fc region having a mutation at position 228. In a further embodiment, the amino acid at position 228 is mutated from serine (S) to proline (P) ( / .e., S228P). In another embodiment, the CB1 antibody exhibits reduced effector function and comprises an lgG2 Fc region having a mutation at position 330 and / or 331. In a further embodiment, the amino acid at position 330 is mutated from alanine (A) to serine (S), and / or the amino acid at position 331 is mutated from proline (P) to serine (S). In a further embodiment, the CB1 antibody comprises an lgG2 Fc domain having both A330S and P331S mutations. In another embodiment, the CB1 antibody comprises an lgG2 / lgG4 hybrid Fc region. For example, in some embodiments, the CB1 antibody comprises a CH1 and hinge region derived from lgG2, and a CH2 and CH3 region derived from lgG4.

[0158] In some embodiments, the VH region of the anti-CB1 antibody or the antigen binding fragment thereof comprises an amino acid sequence that is or is at least 90% identical to SEQ I D NO: 421. In embodiments, the VH is 90% identical, 91% identical, 92% identical, 93% identical, 94% identical, 95% identical, 96% identical, 97% identical, 98% identical, or 99% identical to SEQ ID NO: 421. In some embodiments, the VL region of the anti-CB1 antibody or the antigen binding fragment thereof comprises an amino acid sequence that is or is at least 90% identical to SEQ ID NO: 432. In embodiments, the VL is 90% identical, 91% identical, 92% identical, 93% identical, 94% identical, 95% identical, 96% identical, 97% identical, 98% identical, or 99% identical to SEQMBHB Ref. No.: 24-0691 -WOID NO: 422. In some embodiments, the anti-CB1 antibody or the antigen binding fragment thereof comprises a heavy chain comprising an amino acid sequence that is or is at least 90% identical to SEQ ID NO: 422. In embodiments, the heavy chain is 90% identical, 91% identical, 92% identical, 93% identical, 94% identical, 95% identical, 96% identical, 97% identical, 98% identical, or 99% identical to SEQ ID NO: 421. In some embodiments, the anti-CB1 antibody or the antigen binding fragment thereof comprises a light chain variable region comprising an amino acid sequence that is at least 90% identical to SEQ ID NO: 435. In embodiments, the light chain is 90% identical, 91% identical, 92% identical, 93% identical, 94% identical, 95% identical, 96% identical, 97% identical, 98% identical, or 99% identical to SEQ ID NO: 422.

[0159] In some embodiments, the heavy chain variable region comprises, consists essentially of, or consists of an amino acid sequence selected from the group consisting of SEQ ID NOs: 431-433. In some embodiments, the isolated antibody or antigen binding fragment thereof comprises a heavy chain variable region amino acid sequence that is at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% identical to an amino acid sequence selected from the group consisting of SEQ ID NOs: 431-433. In a further embodiment, the heavy chain variable region comprises, consists essentially of, or consists of an amino acid sequence selected from the group consisting of SEQ ID NOs: 431-433.

[0160] In another embodiment, the isolated antibody or antigen binding fragment thereof comprises a heavy chain amino acid sequence that is least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% identical to an amino acid sequence selected from the group consisting of SEQ ID NO: 435-443. In a further embodiment, the heavy chain comprises, consists essentially of, or consists of an amino acid sequence selected from the group consisting of SEQ ID NO: 435-443.

[0161] In another embodiment, the isolated antibody or antigen binding fragment thereof comprises a light chain variable region amino acid sequence that is least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% identical to an amino acid sequence according to SEQ ID NO: 421. In a further embodiment, the heavy chain variable region comprises, consists essentially of, or consists of an amino acid sequence according to SEQ ID NO: 421. In another embodiment, the isolated antibody or antigen binding fragment thereof comprises a light chain amino acid sequence that is least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% identical to an amino acidMBHB Ref. No.: 24-0691 -WOsequence according to SEQ ID NO: 422. In a further embodiment, the light chain comprises, consists essentially of, or consists of an amino acid sequence according to SEQ ID NO: 422.

[0162] In some embodiments, the disclosure provides a humanized isolated antibody or antigen binding fragment thereof that binds CB1. In a further embodiment, the isolated antibody or antigen binding fragment thereof comprises a light chain variable region according to SEQ ID NO: 421 and a heavy chain variable region according to SEQ ID NO: 431. In another embodiment, the isolated antibody or antigen binding fragment thereof comprises a light chain variable region according to SEQ ID NO: 421 and a heavy chain variable region according to SEQ ID NO: 432. In another embodiment, the isolated antibody or antigen binding fragment thereof comprises a light chain variable region according to SEQ ID NO: 421 and a heavy chain variable region according to SEQ ID NO: 433. In another embodiment, the isolated antibody or antigen binding fragment thereof comprises a full light chain according to SEQ ID NO: 422 and a full heavy chain according to a sequence selected from the group consisting of SEQ ID NOs: 435-443.

[0163] In other embodiments, the isolated antibody or antigen binding fragment thereof comprises a light chain sequence variable region sequence according to one of SEQ ID NO: 423, 425, 427 or 429, or a full-length light chain sequence according to one of SEQ ID NO: 424, 426, 428, or 430. In embodiments, antibody according to the disclosure includes the light chain variable regions is one of one of SEQ ID NO: 423, 425, 427 or 429 and a heavy chain variable region including SEQ ID NO: 431-433. In some embodiments, the isolated antibody or antigen binding fragments comprise a light chain sequence according to SEQ ID NO: 424, 426, 428, or 430, and a heavy chain sequence according to SEQ ID NO: 435-443.

[0164] In some embodiments, the isolated antibody or antigen binding fragment thereof binds CB1 and exhibits reduced effector function. In some embodiments, the isolated antibody or antigen binding fragment thereof binds CB1 and comprises one or more Fc region modifications. In a further embodiment, the antibody or antigen binding fragment thereof binds CB1 and comprises an amino acid sequence comprising one or more mutations in the Fc region. In a further embodiment, the isolated antibody or antigen binding fragment thereof has a mutation at position 228 and / or 330 and / or 331. In another embodiment, the isolated antibody or antigen binding fragment thereof has a mutation at position 228 of the Fc region, wherein the Fc region is of the lgG4 isotype. In a further embodiment, the mutation is S228P. In another embodiment, the isolated antibody or antigen binding fragment thereof has a mutation at position 330 and / or position 331. In a further embodiment, the isolated antibody or antigen binding fragment thereof has a mutation at position 330 and / or 331, wherein the Fc region is of the lgG2 isotype. In a furtherMBHB Ref. No.: 24-0691 -WOembodiment, the isolated antibody or antigen binding fragment thereof has the following mutations in the Fc region: A330S and P331S. In another embodiment, the isolated antibody or antigen binding fragment thereof comprises an Fc region that is a hybrid Fc region. For example, in some embodiments, the Fc region is a hybrid lgG2 / lgG4 Fc region, wherein the CH1 and hinge regions are derived from lgG2, and the CH2 and CH3 regions are derived from lgG4.

[0165] Thus, in some embodiments, the antibody or antigen binding fragment thereof provided herein is a chimeric or humanized antibody or fragment containing heavy and light chain CDRs selected from the CDRs provided herein, or conservative variants of the CDRs provided herein, wherein the isolated antibody or fragment thereof comprises an Fc region comprising modifications that alter antibody effector functions. For example, in some embodiments, the isolated antibody or antigen binding fragment thereof comprises light and heavy chain CDRs according to disclosure, and further comprises an lgG2-lgG4 hybrid Fc region, an lgG2 Fc region comprising amino acid mutations at positions 330 and 331 (e.g., A330S and P331S), or an lgG4 Fc region comprising an amino acid mutation at position 228 (e.g., S228P).

[0166] In the various embodiments of the disclosure, the anti-CB1 antibodies include one or more of the sequences identified in Table 4.Table 4Anti-CB1 Antibody SequencesMBHB Ref. No.: 24-0691 -WOMBHB Ref. No.: 24-0691 -WOMBHB Ref. No.: 24-0691 -WOMBHB Ref. No.: 24-0691 -WO

[0167] In some embodiments, the present disclosure provides an isolated antibody or antigen binding fragment thereof that binds to CB1 , wherein the antibody or fragment has a binding affinity Kd for CB1 of about 70 nM or less, about 60 nM or less, about 50 nM or less, about 40 nM or less, about 30 nM or less, about 25 nM or less, about 20 nM or less, about 15 nM or less, about 10 nM or less, about 8 nM or less, about 6 nM or less, about 5 nM or less, about 4 nM or less, about 3 nM or less, about 2 nM or less, or about 1 nM or less. In some embodiments, present disclosure provides an isolated antibody or fragment thereof that binds to CB1 , wherein the antibody or fragment has a binding affinity Kd forCBI in the range of about 1nM to about 100 nM, about 2 nM to about 75 nM, about 3 nM to about 50 nM, about 4 nM to about 10 nM, or has a binding affinity Kd for CB2 receptor that is about 50 nM, or about 40 nM, or about 30 nM, or about 20 nM, or about 10 nM, or about 5 nM, or about 4 nM, or about 3 nM or about 2 nM, or about 1 nM.

[0168] In some embodiments, the present disclosure provides an isolated antibody or antigen binding fragment thereof that is at least 2 fold, at least 3 fold, at least 4 fold, at least 5 fold, at least 6 fold, at least 7 fold, at least 8 fold, at least 9 fold, at least 10 fold, at least 11 fold, at least 12 fold, at least 13 fold, at least 14 fold, or at least 15 fold more potent than the small molecule rimonabant, wherein the potency of the antibody or fragment or rimonabant is measured byMBHB Ref. No.: 24-0691 -WOinhibition of CB1 antagonist-mediated signal transduction in a cAMP assay. In a further embodiment, the isolated antibody or antigen binding fragment thereof is humanized.

[0169] In some embodiments, the present disclosure provides an isolated humanized antibody or antigen binding fragment thereof that binds to CB1 , wherein the antibody or fragment exhibits greater binding affinity and / or greater potency than a corresponding non-humanized or chimeric antibody, wherein the humanized antibody or fragment and the corresponding nonhumanized or chimeric antibody comprise the same heavy and light chain CDRs. For example, in some embodiments, the present disclosure provides a humanized antibody or fragment thereof comprising heavy chain CDR1, CDR2, and CDR3 and light chain CDR1, CDR2, and CDR3 according to the disclosure, wherein the humanized antibody exhibits greater binding affinity for CB1 and / or greater potency with respect to inhibition of a CB1 agonist. In some embodiments, the humanized antibodies and fragments provided herein exhibit at least 50% greater, at least 100% greater, at least 2 fold greater, at least 3 fold greater, at least 4 fold greater, at least 5 fold greater, or at least 10 fold greater potency relative to the corresponding non-humanized or chimeric antibody. In a further embodiment, the potency is measured by inhibition of CB1-cAMP production.

[0170] Potency of anti-CB1 antibodies provided herein may be measured by any method known in the art. For example, in some embodiments, potency of the antibodies and fragments provided herein is measured by intracellular cAMP levels or ERK phosphorylation. For example, potency may be measured by the level of inhibition cAMP production in a cAMP functional assay (Cisbio) or inhibition of WIN55, 212-induced ERK phosphorylation in a Western blot.Indications

[0171] As used herein, the term “obesity” is based on BMI for both youth and adults, but the definitions are not directly comparable. Among adults, there is a set cut point based on health risk, while among children the definition is statistical and is based on a comparison to a reference population. BMI is calculated as weight in kilograms divided by height in meters squared, rounded to one decimal place. Obesity in adults is defined as a BMI of greater than or equal to 30 kg / m2. Obesity in youth is defined as a BMI of greater than or equal to the age- and sex-specific 95th percentile of the 2000 CDC growth charts.

[0172] The term “overweight condition” refers to a body mass index (BMI) greater than or equal to 25 kg / m2and less than 30 kg / m2. Overweight condition may also be associated with atMBHB Ref. No.: 24-0691 -WOleast one additional risk factor for fatal diseases (stroke, Ml, heart failure, sudden death) such as diabetes, hypertension, family history of premature coronary artery disease, etc.).

[0173] The term “type II diabetes”, also referred to as type 2 diabetes, previously referred to as "non-insulin-dependent diabetes" or "adult-onset diabetes," accounts for 90-95% of all diabetes, encompasses individuals who have insulin resistance and usually relative (rather than absolute) insulin deficiency. At least initially, and often throughout their lifetime, these individuals may not need insulin treatment to survive. There are various causes of type 2 diabetes. Although the specific etiologies are not known, autoimmune destruction of B-cells does not occur, and patients do not have any of the other known causes of diabetes. Most, but not all, patients with type 2 diabetes are overweight or obese. Excess weight itself causes some degree of insulin resistance. Patients who are not obese or overweight by traditional weight criteria may have an increased percentage of body fat distributed predominantly in the abdominal region. Type 2 diabetes frequently goes undiagnosed for many years because hyperglycemia develops gradually and, at earlier stages, is often not severe enough for the patient to notice the classic diabetes symptoms. Nevertheless, even undiagnosed patients are at increased risk of developing macrovascular and microvascular complications.

[0174] The term “comorbidities” of obesity or being overweight means other diseases, disorders, or syndromes that are associated with, accompany, or are caused by being obese or overweight. Obesity is associated with serious chronic disorders including but not limited to type II diabetes or glucose intolerance, prediabetes, high triglycerides, physical impairment, osteoporosis, renal disease, obstructive sleep apnea, sexual hormones impairment, endocrine reproductive disorders such as polycystic ovary syndrome or male hypogonadism, osteoarthritis, gastrointestinal cancers, dyslipidemia, hypertension, heart failure, coronary heart disease and stroke, gallstones, hypertension, and an altered gonadal hormone profile. “Glucose intolerance” is defined as the inability to properly metabolize glucose.

[0175] “Insulin sensitivity” describes how sensitive the body is to the effects of insulin. Someone said to be insulin sensitive will require smaller amounts of insulin to lower blood glucose levels than someone who has low sensitivity. Insulin sensitivity varies from person to person and doctors can perform tests to determine how sensitive an individual is to insulin.

[0176] Herein the terms "improving insulin sensitivity" and "treating / lowering insulin resistance" shall be construed as equivalent.MBHB Ref. No.: 24-0691 -WO

[0177] Many treatments have been developed and are in clinical use for type II diabetes. Each of these can be used alongside the antibodies and conjugates thereof disclosed herein. Each of these can be used alongside the ADCs disclosed herein. Examples of such treatments include: Biguanides

[0178] Examples of biguanides include metformin, phenformin, and buformin. Generally, metformin is the first medication prescribed for type II diabetes. It works by improving the sensitivity of body tissues to insulin so that the body uses insulin more effectively. Metformin also lowers glucose production in the liver. Metformin may not lower blood sugar enough on its own. DPP-4 inhibitors

[0179] DPP-4 inhibitors inhibit the enzyme DPP-4 (dipeptidyl peptidase-4). This protease degrades incretins, a class of proteins that serve as metabolic hormones and stimulate a decrease in blood glucose levels. Circulating levels of incretins increase after a meal and supplement the insulin released from pancreatic beta cells as a response to blood glucose levels. DPP-4 inhibitors thereby reduce blood sugar levels. They do not cause weight gain. Examples of these inhibitors include sitagliptin, saxagliptin, vildagliptin, linagliptin, gemigliptin, anagliptin, tenegliptin, alogliptin, omarigliptin, evogliptin, gosogliptin, dutogliptin, neogliptin, retagliptin, denagliptin, cofrogliptin, fotagliptin, prusogliptin, berberine, among others.Sulfonylureas

[0180] These medications increase insulin secretion from pancreatic beta cells. Examples of medications in this class include acetohexamide, carbutamide, chlorpropamide, glycyclamide (tolcyclamide), metahexamide, tolazamide tolbutamide, glibenclamide (glyburide), glibornuride, gliclazide, glipizide, gliquidone, glisoxepide glyclopyramide, and glimepiride.Meglitinides

[0181] These drugs bind to an ATP-dependent potassium (KATP) channel on the cell membrane of pancreatic beta cells in a similar manner to sulfonylureas, albeit with weaker binding affinity and faster dissociation from the binding site. Drug binding increases the concentration of intracellular potassium, causing the electric potential toward the intracellular side of the membrane to become more positive and inducing depolarization. This depolarization opens voltage-gated Ca2+channels, stimulating a rise in intracellular calcium leading to increased fusion of insulin granula in the cell membrane and therefore increased secretion of insulin precursorMBHB Ref. No.: 24-0691 -WOproinsulin. Examples include meglitinide, repaglinide, nateglinide, and mitiglinide. Meglitinides are also known as glinides.Thiazolidinediones

[0182] Like metformin, these medications make the body's tissues more sensitive to insulin. Thiazolidinediones are also known as TZDs or glitazones. TZDs act by activating PPARs (peroxisome proliferator-activated receptors), a group of nuclear receptors, specific for PPARy (PPAR-gamma, PPARG). The endogenous ligands for these receptors are free fatty acids (FFAs) and eicosanoids. When activated, the receptor binds to DNA in complex with the retinoid X receptor (RXR), another nuclear receptor, increasing transcription of a number of specific genes and decreasing transcription of others. The main effect of expression and repression of these specific genes is an increase in the storage of fatty acids in adipocytes, thereby decreasing the amount of fatty acids present in circulation. As a result, cells become more dependent on the oxidation of carbohydrates, more specifically glucose, in order to yield energy for other cellular processes, which promotes glucose uptake that is normally deficient in type II diabetes (Eggleton and Jialal, “Thiazolidinediones”. In: StatPearls [Internet], Treasure Island (FL): StatPearls Publishing; 2024, PMID: 31869120). Examples of thiazolidinediones include pioglitazone, rosiglitazone, lobeglitazone, azemiglitazone, ciglitazone, darglitazone, englitazone, netoglitazone, rivoglitazone, troglitazone, balaglitazone (DRF-2593), and AS-605240.SGLT2 inhibitors

[0183] Inhibitors of the sodium / glucose cotransporter 2 (SGLT2), which is specifically expressed in nephrons, work by preventing the kidneys from reabsorbing glucose into the blood. Instead, excess glucose is excreted into the urine, which lowers blood glucose levels. Aside from blood sugar control, SGLT2 inhibitors have been shown to provide significant cardiovascular benefit in people with type II diabetes, causing a reduction in body weight as well as systolic and diastolic blood pressure (Usman et al., 2018, Eur. J. Prev. Cardiol. 25(5): 495-502). SGLT2 inhibitors are also called gliflozins or flozins. Examples include bexagliflozin, canagliflozin, dapagliflozin, empagliflozin, ertugliflozin, ipragliflozin, luseogliflozin, remogliflozin etabonate, sergliflozin etabonate, sotagliflozin, tofogliflozin, and henagliflozin, as well as salts thereof.Insulin therapy

[0184] Some patients with type II diabetes require insulin therapy as well. Because normal digestion interferes with insulin taken by mouth, insulin must be injected. Depending on needs, a mixture of insulin types may be used throughout the day and night. Often, people with type 2MBHB Ref. No.: 24-0691 -WOdiabetes start insulin use with one long-acting shot at night. There are many types of insulin, and they each work in a different way. Many of these types of insulin are modified or derivatized to impart favorable properties, such as an extended in vivo half-life. Examples include insulin glulisine, insulin lispro, insulin aspart, NPH insulin, insulin degludec, insulin glargine, insulin detemir, and insulin isophane.

[0185] Other drugs that can be used in combination with the antibodies and ADCs described herein include insulin sensitivity enhancing agents, alpha-glucosidase inhibitors, biguanides, amyrin agonists, phosphotyrosin phosphatase (PTP) inhibitors, aldose reductase inhibitors, neurotrophic factors, PKC inhibitors, advanced glycation end-product (AGE) inhibitors, active oxygen quenching agents, statins, squalene synthetase inhibitors, fibrate, niacin, PCSK9 inhibitors, triglyceride lowing agents, cholesterol sequestering agents, angiotensin converting enzyme inhibitors, angiotensin II antagonists, calcium channel blockers, ursodiol, orlistat, betaine, CNS-acting anti-obesity agents, gastrointestinal lipase inhibitors, p3-adrenoceptor agonists, peptide-based appetite-suppressing agents, cholecystokinin agonists, dopamine agonists, other glucagon-like peptides, meglitinides, cyclooxygenase inhibitors, progesterone derivatives, metoclopramide-based agents, tetrahydrocannabinol-based agents, and lipid metabolism improving agents.

[0186] Each of the above diabetes treatments can be used in combination with the antibodies and ADCs described herein. The treatments can be administered simultaneously, separately, or sequentially.CB1 (cannabinoid receptor 1) and metabolic diseases

[0187] Cannabinoid receptor 1 (CB1) is a member of the G protein-coupled receptor (GPCR) superfamily. CB1 is expressed in the central nervous system (CNS), lungs, liver, adipose tissue and kidneys, and has been implicated in many human diseases including obesity, diabetes, fibrosis, liver diseases, cardiovascular disease, cancer, pain, MS spasticity, and glaucoma, among others. More specifically, CB1 has been shown to exhibit detrimental activity in, for example, obesity, diabetes, fibrosis, liver diseases, cardiovascular disease and cancer; and has been shown to exhibit beneficial activity in pain, MS spasticity and glaucoma, among others (Kunos et al., 2009, Trends Pharmacol Sci 30:1-7).

[0188] There is a need in the art for new CB1 antagonists and agonists for therapeutic purposes as well as selective binders for diagnostic / imaging purposes. In particular, a CB1-MBHB Ref. No.: 24-0691 -WOtargeting compound that lacks the capacity for CNS penetration would be desirable to reduce potential CNS-mediated side effects of CB1 modulation, highlighted by the psychiatric adverse events associated with the CB1 inverse agonist rimonabant, which was effective in reducing body weight and improving insulin resistance and dyslipidemia in overweight patients but was associated with emergent depression and anxiety in a small subset of patients (Kunos and Tam, 2011, Br. J. Pharmacol. 163(7): 1423-1431). CB1 blockade using rimonabant reverses inflammation-induced impairment of glucose uptake in adipocytes isolated from high-fat diet rats (Miranville et al, 2010, Obesity 18: 2247-2254). Rimonabant was additionally shown to improve the profile of several metabolic risk factors (including adiponectin levels) in overweight patients according to the Rimonabant in Obesity-Lipids (RIO-Lipids) study. (See e.g., Despres etal., 2005, N Engl J Med, 353:2121-2134).

[0189] As described herein, the term “metabolic disease” can mean type I diabetes, type II diabetes, obesity, metabolic syndrome / pre-diabetes, cardiovascular disease, non-alcoholic steatohepatitis (NASH), spinal cord injury (SCI), a hypo-metabolic state, double diabetes, Cushings disease, and an obesity syndrome associated with a hypothalamic disorder. In some embodiments, the human subject is an adult human subject. In some embodiments, the human subject is a pediatric human subject. In some embodiments, the subject is treated with a second therapy as described herein.

[0190] When discussing a pharmaceutical intervention, the term “treatment” is synonymous with drug, drug therapy, therapeutic, therapeutic regimen, course of treatment, and pharmaceutical agent. The use of these terms to describe medical interventions is well known to those in the art.

[0191] The terms “administer”, “administering”, “administration” include any method of delivery of an antibody, antigen-binding fragment thereof, or antibody-drug conjugate, e.g., a pharmaceutical composition comprising such an antibody, antigen-binding fragment, or antibodydrug conjugate, into a subject’s system or to a particular region in or on a subject (known as systemic and local administration, respectively).

[0192] In some embodiments, an “effective amount” is an amount that, when administered according to a particular regimen, produces a positive clinical outcome with a reasonably acceptable level of adverse effects (e.g., toxicity), such that the adverse effects, if present, are tolerable enough for a patient to continue with the therapeutic regimen, and the benefit of the therapy overweighs risk of toxicity. Those of ordinary skill in the art will appreciate that in some embodiments of the invention, a unit dosage may be considered to contain an effective amount ifMBHB Ref. No.: 24-0691 -WOit contains an amount appropriate for administration in the context of a dosage regimen correlated with a positive outcome. A therapeutically effective amount is commonly administered in a dosing regimen that may comprise multiple unit doses. For any particular pharmaceutical agent, a therapeutically effective amount (and / or an appropriate unit dose within an effective dosing regimen) may vary, for example, depending on route of administration, on combination with other pharmaceutical agents. In some embodiments, the specific therapeutically effective amount (and / or unit dose) for any particular patient may depend upon a variety of factors including the disorder being treated and the severity of the disorder; the activity of the specific pharmaceutical agent employed; the specific composition employed; the age, body weight, general health, sex and diet of the patient; the time of administration, route of administration, and / or rate of excretion or metabolism of the specific pharmaceutical agent employed; the duration of the treatment; and similar factors known in the medical arts.

[0193] By “treating” or “preventing” a disease or disorder is meant delaying or preventing the onset of such a disease or disorder, reversing, alleviating, ameliorating, inhibiting, slowing down or stopping the progression, aggravation or deterioration, the progression or severity of a condition associated with such a disease or disorder, but not necessarily requiring a complete treatment or prevention of the disease or disorder. In some embodiments, the symptoms of a disease or disorder are alleviated by at least 5%, at least 10%, at least 20%, at least 30%, at least 40%, or at least 50%.

[0194] CB1 antagonists, particularly small molecules, are associated with neuropsychiatric side effects as a result of their antagonism of CB1 in the central nervous system (CNS). These side effects can limit their utility for treatment of metabolic diseases. In contrast, prior work has shown that semaglutide cannot cross the blood-brain barrier (Gabery etal., 2020, JCI Insight 5(6): e133429), and it is generally known that most ADCs are also unable to cross the blood-brain barrier without significant modifications to enable penetration into the CNS (see, for example, Cavaco et al., 2021, ACS Med. Chem. Lett. 12(11): 1663-1668). Thus, combining the effectiveness of anti-CB1 antibodies for metabolic disease with (1) the effectiveness and periphery-specific action of semaglutide and (2) the peripheral tissue bias of ADC technology could enable more effective treatment of metabolic disease at lower doses without the side effects traditionally associated with anti-CB1 therapies.

[0195] In one aspect, the anti-CB1 antibodies, or conjugates thereof, disclosed herein are useful for antagonizing CB1 activity. Accordingly, in another aspect, the invention provides methods for treating CB1 -associated diseases or disorders by administering to a subject in needMBHB Ref. No.: 24-0691 -WOof thereof a pharmaceutical composition comprising one or more anti-CB1 antibodies or conjugates, or antigen binding fragments thereof disclosed herein. In some embodiments, the antagonistic anti-CB1 antibodies and conjugates thereof provided herein provide a beneficial effect when used as a treatment for, or for prevention of, obesity, diabetes, fibrosis, liver diseases, cardiovascular diseases, or other metabolic diseases. In some embodiments, the antagonistic anti-CB1 antibodies and conjugates provided herein are therapeutically beneficial for treating or preventing a disease mediated by CB1 activity.

[0196] In some embodiments, the invention comprises a pharmaceutical composition comprising the antibody or antibody-drug conjugate described herein. The pharmaceutical composition can comprise the antibody or conjugate along with a pharmaceutically acceptable carrier, diluent, excipient, buffer, or the like.

[0197] As used herein, an “excipient” is an inactive ingredient in a pharmaceutical composition. Examples of excipients include fillers or diluents, surfactants, binders, glidants, lubricants, colorants, disintegrants, and the like.

[0198] As used herein, a “disintegrant” is an excipient that hydrates a pharmaceutical composition and aids in tablet dispersion. Examples of disintegrants include sodium croscarmellose and / or sodium starch glycolate.

[0199] As used herein, a “diluent” or “filler” is an excipient that adds bulkiness to a pharmaceutical composition. Examples of fillers include lactose, sorbitol, celluloses, calcium phosphates, starches, sugars (e.g., mannitol, sucrose, or the like) or any combination thereof.

[0200] As used herein, a “surfactant” is an excipient that imparts pharmaceutical compositions with enhanced solubility and / or wettability. Examples of surfactants include sodium lauryl sulfate (SLS), sodium stearyl fumarate (SSF), polyoxyethylene 20 sorbitan mono-oleate (e.g., Tween™), or any combination thereof.

[0201] As used herein, a “binder” is an excipient that imparts a pharmaceutical composition with enhanced cohesion or tensile strength (e.g., hardness). Examples of binders include dibasic calcium phosphate, sucrose, corn (maize) starch, microcrystalline cellulose, and modified cellulose (e.g., hydroxymethyl cellulose).

[0202] As used herein, a “glidant” is an excipient that imparts a pharmaceutical composition with enhanced flow properties. Examples of glidants include colloidal silica and / or talc.MBHB Ref. No.: 24-0691 -WO

[0203] As used herein, a “colorant” is an excipient that imparts a pharmaceutical composition with a desired color. Examples of colorants include commercially available pigments such as FD&C Blue #1 Aluminum Lake, FD&C Blue #2, other FD&C Blue colors, titanium dioxide, iron oxide, and / or combinations thereof.

[0204] As used herein, a “lubricant” is an excipient that is added to pharmaceutical compositions that are pressed into tablets. The lubricant aids in compaction of granules into tablets and ejection of a tablet of a pharmaceutical composition from a die press. Examples of lubricants include magnesium stearate, stearic acid (stearin), hydrogenated oil, sodium stearyl fumarate, or any combination thereof.

[0205] As used herein, a “carrier” can comprise a polymer (e.g., a water-soluble polymer or a partially water-soluble polymer) and can include optional excipients such as functional excipients (e.g., one or more surfactants) or nonfunctional excipients (e.g., one or more fillers).

[0206] As used herein, the term “buffer” can refer to any solution with a controlled pH that may serve to dissolve a solid (e.g., lyophilized) pharmaceutical or as a diluent to dilute a liquid pharmaceutical.

[0207] In one embodiment, at least one of the peptides SBI-401 (SEQ ID NO: 2), SBI-403 (SEQ ID NO: 4), SBI-404 (SEQ ID NO: 5), SBI-405 (SEQ ID NO: 6), SBI-406 (SEQ ID NO: 7), SBI-407 (SEQ ID NO: 8), SBI-408 (SEQ ID NO: 9), SBI-409 (SEQ ID NO: 10), SBI-410 (SEQ ID NO: 11), and SBI-411 (SEQ ID NO: 12) is conjugated to an isolated antibody or an antigen binding fragment thereof that specifically binds to human cannabinoid 1 (CB1) receptor.

[0208] In various embodiments, the anti-CB1 antibodies for use in preparation of conjugates according to the disclosure include: an isolated antibody or antigen binding fragment thereof comprising: (i) a heavy chain variable region (“VH” or “VH region”) comprising the following complementarity determining regions (CDRs): HCDR1, HDCR2, and HCR3 each having one of the amino acid sequences identified in Table X, including for example an HCDR1 of SEQ ID NO: 26; HCDR2 having the amino acid sequence of SEQ ID NO: 27; HCDR3 having the amino acid sequence of SEQ ID NO: 28; and (ii) a light chain variable region (“VL” or “VL region”) comprising the following CDRs: LCDR1, LCR2, and LCDR3 each having one the amino acid sequences identified in Table X, including for example an LCDR1 having the amino acid sequence of SEQ ID NO: 29; LCDR2 having the amino acid sequence of SEQ ID NO: 30; LCDR3 having the amino acid sequence of SEQ ID NO: 31; or a pharmaceutically acceptable salt, solvate, stereoisomer, or tautomer thereof; wherein the VH region of the isolated antibody or the antigen bindingMBHB Ref. No.: 24-0691 -WOfragment thereof comprises an amino acid sequence that is 90% identical to SEQ ID NO: 32; or wherein the VL region of the isolated antibody or the antigen binding fragment thereof comprises an amino acid sequence that is 90% identical to SEQ ID NO: 33; or wherein the isolated antibody or the antigen binding fragment thereof comprises: (a) a heavy chain comprising an amino acid sequence at least 90% identical to SEQ ID NO: 32; and (b) a light chain variable region comprising an amino acid sequence that is at least 90% identical to SEQ ID NO: 33; and wherein the isolated antibody or the antigen binding fragment thereof to the at least one of the peptides disclosed herein ratio is less than 1; and wherein the at least one of the peptides disclosed herein is linked to the isolated antibody or the antigen binding fragment thereof via cysteine- based bioconjugation or the at least one of the peptides disclosed herein is linked site-specifically to the isolated antibody or the antigen binding fragment thereof or the at least one of the peptides disclosed herein is conjugated to the N-terminus of the isolated antibody or the antigen binding fragment or the at least one of the peptides disclosed herein is conjugated to the C-terminus of the isolated antibody or the antigen binding fragment or the at least one of the peptides disclosed herein is conjugated to the Fc region of the isolated antibody or the antigen binding fragment.

[0209] In another embodiment, the conjugate comprising at least one of the peptides SBI-401 (SEQ ID NO: 2), SBI-403 (SEQ ID NO: 4), SBI-404 (SEQ ID NO: 5), SBI-405 (SEQ ID NO: 6), SBI-406 (SEQ ID NO: 7), SBI-407 (SEQ ID NO: 8), SBI-408 (SEQ ID NO: 9), SBI-409 (SEQ ID NO: 10), SBI-410 (SEQ ID NO: 11), and SBI-411 (SEQ ID NO: 12) is conjugated to an isolated antibody or an antigen binding fragment thereof that specifically binds to human cannabinoid 1 (CB1) receptor; wherein isolated antibody or antigen binding fragment thereof comprises: (i) a heavy chain variable (VH) region comprising the following complementarity determining regions (CDRs): HCDR1 having the amino acid sequence of SEQ ID NO: 26; HCDR2 having the amino acid sequence of SEQ ID NO: 27; HCDR3 having the amino acid sequence of SEQ ID NO: 28; and (ii) a light chain variable (VL) region comprising the following CDRs: LCDR1 having the amino acid sequence of SEQ ID NO: 29; LCDR2 having the amino acid sequence of SEQ ID NO: 30; LCDR3 having the amino acid sequence of SEQ ID NO: 31; or a pharmaceutically acceptable salt, solvate, stereoisomer, or tautomer thereof; wherein the VH region of the isolated antibody or the antigen binding fragment thereof comprises an amino acid sequence that is 90% identical to SEQ ID NO: 32; or wherein the VL region of the isolated antibody or the antigen binding fragment thereof comprises an amino acid sequence that is 90% identical to SEQ ID NO: 33; or wherein the isolated antibody or the antigen binding fragment thereof comprises: (a) a heavy chain comprising an amino acid sequence at least 90% identical to SEQ ID NO: 32; and (b) a light chain variable region comprising an amino acid sequence that is at least 90% identical to SEQ ID NO:MBHB Ref. No.: 24-0691 -WO33; and wherein the isolated antibody or the antigen binding fragment thereof to the at least one of the peptides disclosed herein ratio is less than 1; and wherein the at least one of the peptides disclosed herein is linked to the isolated antibody or the antigen binding fragment thereof via cysteine-based bioconjugation or the at least one of the peptides disclosed herein is linked site-specifically to the isolated antibody or the antigen binding fragment thereof or the at least one of the peptides disclosed herein is conjugated to the N-terminus of the isolated antibody or the antigen binding fragment or the at least one of the peptides disclosed herein is conjugated to the C-terminus of the isolated antibody or the antigen binding fragment or the at least one of the peptides disclosed herein is conjugated to the Fc region of the isolated antibody or the antigen binding fragment is used in a pharmaceutical composition, further comprising a pharmaceutically acceptable excipient, carrier, or diluent.

[0210] In another embodiment, the pharmaceutical composition comprising the conjugate comprising at least one of the peptides SBI-401 (SEQ ID NO: 2), SBI-403 (SEQ ID NO: 4), SBI-404 (SEQ ID NO: 5), SBI-405 (SEQ ID NO: 6), SBI-406 (SEQ ID NO: 7), SBI-407 (SEQ ID NO: 8), SBI-408 (SEQ ID NO: 9), SBI-409 (SEQ ID NO: 10), SBI-410 (SEQ ID NO: 11), and SBI-411 (SEQ ID NO: 12) is conjugated to an isolated antibody or an antigen binding fragment thereof that specifically binds to human cannabinoid 1 (CB1) receptor; wherein isolated antibody or antigen binding fragment thereof comprises: (i) a heavy chain variable (VH) region comprising the following complementarity determining regions (CDRs): HCDR1 having the amino acid sequence of SEQ ID NO: 26; HCDR2 having the amino acid sequence of SEQ ID NO: 27; HCDR3 having the amino acid sequence of SEQ ID NO: 28; and (ii) a light chain variable (VL) region comprising the following CDRs: LCDR1 having the amino acid sequence of SEQ ID NO: 29; LCDR2 having the amino acid sequence of SEQ ID NO: 30; LCDR3 having the amino acid sequence of SEQ ID NO: 31; ora pharmaceutically acceptable salt, solvate, stereoisomer, or tautomer thereof; wherein the VH region of the isolated antibody or the antigen binding fragment thereof comprises an amino acid sequence that is 90% identical to SEQ ID NO: 32; or wherein the VL region of the isolated antibody or the antigen binding fragment thereof comprises an amino acid sequence that is 90% identical to SEQ ID NO: 33; or wherein the isolated antibody or the antigen binding fragment thereof comprises: (a) a heavy chain comprising an amino acid sequence at least 90% identical to SEQ ID NO: 32; and (b) a light chain variable region comprising an amino acid sequence that is at least 90% identical to SEQ ID NO: 33; and wherein the isolated antibody or the antigen binding fragment thereof to the at least one of the peptides disclosed herein ratio is less than 1; and wherein the at least one of the peptides disclosed herein is linked to the isolated antibody or the antigen binding fragment thereof via cysteine- based bioconjugation or the at least one of theMBHB Ref. No.: 24-0691 -WOpeptides disclosed herein is linked site-specifically to the isolated antibody or the antigen binding fragment thereof or the at least one of the peptides disclosed herein is conjugated to the N-terminus of the isolated antibody or the antigen binding fragment or the at least one of the peptides disclosed herein is conjugated to the C-terminus of the isolated antibody or the antigen binding fragment or the at least one of the peptides disclosed herein is conjugated to the Fc region of the isolated antibody or the antigen binding fragment is used in a pharmaceutical composition, and further comprising a pharmaceutically acceptable excipient, carrier, or diluent is used in a method for treating obesity and / or type 2 diabetes in a subject, the method comprising administering to the subject in need of such treatment an effective amount the pharmaceutical composition.

[0211] In another embodiment, the conjugate comprising at least one of the peptides SBI-401 (SEQ ID NO: 2), SBI-403 (SEQ ID NO: 4), SBI-404 (SEQ ID NO: 5), SBI-405 (SEQ ID NO: 6), SBI-406 (SEQ ID NO: 7), SBI-407 (SEQ ID NO: 8), SBI-408 (SEQ ID NO: 9), SBI-409 (SEQ ID NO: 10), SBI-410 (SEQ ID NO: 11), and SBI-411 (SEQ ID NO: 12) is conjugated to an isolated antibody or an antigen binding fragment thereof that specifically binds to human cannabinoid 1 (CB1) receptor or a pharmaceutically acceptable salt, solvate, stereoisomer, or tautomer thereof; wherein the isolated antibody or the antigen binding fragment thereof to the at least one of the peptides disclosed herein ratio is less than 1; and wherein the at least one of the peptides disclosed herein is linked to the isolated antibody or the antigen binding fragment thereof via cysteine-based bioconjugation or the at least one of the peptides disclosed herein is linked site-specifically to the isolated antibody or the antigen binding fragment thereof or the at least one of the peptides disclosed herein is conjugated to the N-terminus of the isolated antibody or the antigen binding fragment or the at least one of the peptides disclosed herein is conjugated to the C-terminus of the isolated antibody or the antigen binding fragment or the at least one of the peptides disclosed herein is conjugated to the Fc region of the isolated antibody or the antigen binding fragment or the pharmaceutical composition comprising the conjugate is used in a kit. The peptide and antibody components may be provided in isolated, purified, lyophilized, or solution form. The antibody may be a full-length monoclonal antibody, an antibody fragment (e.g., Fab, F(ab’)2, scFv), or an engineered antibody containing one or more conjugation sites.

[0212] In various embodiments, the kit further comprises buffers, reducing agents, activation reagents, catalysts, enzymes, or other auxiliary components that facilitate conjugation, purification, or analysis of the resulting conjugates. Such auxiliary components may include, for example, phosphate-buffered saline, acetate buffers, reducing agents (e.g., TCEP), stabilizers, or quenching reagents.MBHB Ref. No.: 24-0691 -WO

[0213] The kit components may be packaged separately or in combination, optionally in sterile vials, ampoules, or other suitable containers. The kit components may also include instructions for use.

[0214] The disclosure further encompasses kits comprising analytical reagents, standards, or reference materials suitable for determining DAR values or verifying DAR distributions of ADCs produced according to the instructions provided.

[0215] The invention will be further described in the following examples, which do not limit the scope of the invention described in the claims.EXAMPLES

[0216] The Examples that follow are illustrative of specific embodiments of the invention, and various uses thereof. They are set forth for explanatory purposes only and are not to be taken as limiting the invention.Example 1 : General Preparation of ADC Constructs

[0217] This section describes standard solid-phase peptide synthesis protocols, including amino acid deprotection, peptide cleavage from the resin, and final purification. Standard Fmoc-protected amino acids derivatives were employed, unless otherwise noted. Examples include Fmoc-Ala-OH, Fmoc-Arg(Pbf)-OH, Fmoc-Asn(Trt)-OH, Fmoc-Asp(Trt)-OH, Fmoc-Asp(OtBu)-OH, Fmoc-Cys(Acm)-OH, Fmoc-Gln(Trt)-OH, Fmoc-lle-OH, Fmoc-Leu-OH, Fmoc-Lys(Boc)-OH, Fmoc-Met-OH, Fmoc-Phe-OH, Fmoc-Pro-OH, Fmoc-Ser(tBu)-OH, Fmoc-Thr(tBu)-OH, Fmoc-Trp(Boc)-OH, Fmoc-Tyr(tBu)-OH, and Fmoc-Val-OH (all obtained from commercial suppliers such as Novabiochem). Additional Fmoc-protected derivatives used include, but not limited to: Fmoc-Aib-OH, Fmoc-hArg-OH, Fmoc-NMe-Leu-OH, Fmoc-[pl-Phe]-OH, Fmoc-[D-Bip]-OH, Fmoc-Oic-OH, Fmoc-Sar-OH, Fmoc-NH-PEGs-COOH, Fmoc-NH-PEGi2-COOH, and Fmoc-NH-PEG16-COOH. For N-terminal amino acids, Boc-protected building blocks were used. Cysteine-conjugation moieties include 3-Mal-COOH, 6-Mal-COOH, and Br-AcOH.SPPS Method A

[0218] Method A: Solid-phase peptide synthesis is performed on an APEX 396 peptide synthesizer using standard Fmoc chemistry with Rink amide MBHA resin or CTC resin, as follows: Each coupling step is carried out by agitating the resin suspension in DMF with the appropriately protected amino acid derivative, / V,A / '-diisopropylcarbodiimide (DIC), and 1 -hydroxybenzotriazoleMBHB Ref. No.: 24-0691 -WO(CI-HOBt) for 40 min at room temperature. Fmoc deprotection is achieved by treating the resin with 20 % (v / v) piperidine in DMF for 20 min at room temperature. Between each deprotection and coupling cycle, the resin is washed three times with DMF. A ninhydrin test is used to monitor both Fmoc-deprotection and coupling efficiency. If the ninhydrin test indicates incomplete coupling, a second coupling reaction is performed. In the peptide sequences, the s-amino group of lysine residues is protected with either Dde or Boc, depending on the synthetic strategy for attaching cysteine-conjugation moieties or other modified amino acids. Where a cysteine-conjugation moiety (e.g., 3-Mal, 6-Mal, or Br-Ac) is required, it is coupled to the peptide following the removal of the protecting group.Lysine Side-Chain Coupling Method B

[0219] Method B: Modified lysine residues are prepared using Fmoc-Lys(Dde)-OH as the building block. The Dde protecting group is selectively removed with 1 % (v / v) hydrazine in DMF. The resulting c-amino group is then functionalized according to the coupling procedure described in Method A, using the appropriate Fmoc-protected amino acid derivatives or cysteine-conjugation moieties disclosed herein.On-resin Disulfide Bond Formation Method C

[0220] Method C: Prior to the incorporation of cysteine-conjugation moieties (e.g., 3-Mal and 6-Mal in SBI-405 and SBI-406), the peptidyl resin is treated with h in DMF at room temperature to form intramolecular disulfide bonds. Upon completion of the oxidation step, the resin is sequentially washed with DMF (3x), 2% w / w ascorbic acid in DMF (2x), DMF (5x), and DCM (3x). The cysteine-conjugation moieties are then coupled using the procedure described in Method A.Cleavage and Purification Method D

[0221] Method D: The peptide is cleaved from the resin using a cleavage cocktail of TFA / TIS / water (95:2.5:2.5, v / v / v) at room temperature for 1-3 h, the resin is then removed by filtration, and the filtrate is stirred while chilled diethyl ether (pre-cooled in an ice-water bath) is added to precipitate the peptide. The precipitate is collected by centrifugation, redissolved in a suitable solvent (e.g., 50% v / v acetonitrile in water), and purified by preparative RP-HPLC on a C18 column with a mobile phase of acetonitrile / water containing TFA. Fractions are analyzed by HPLC and LCMS; those of appropriate purity are pooled and lyophilized to afford the target peptide.MBHB Ref. No.: 24-0691 -WOExample 2: Preparation of PeptidesPreparation of SB 1-401"<SBI-401 is a GLP-1 analog.Preparation method: SPPS Method A; Lysine Side-Chain Coupling Method B; Cleavage and Purification Method D. Purity: 95.1%; LCMS m / z Calc’d for Ci9iH29oBrN55066 4489.02, found 1498.45 [M + 3H]3+. See FIG. 5A and FIG. 6A.Preparation of SBI-402"" " > SBI-402 is a GLP-1 analog.Preparation method: SPPS Method A; Lysine Side-Chain Coupling Method B; Cleavage and Purification Method D. Purity: 95.6%; LCMS m / z'. Calc’d for C198H312N42O674350.23, found 871.40 [M + 5H]5+. See Fig. 5B and Fig. 6B.Preparation of SBI-403SBI-403 comprises a GLP-1 analog moiety found in albiglutide.MBHB Ref. No.: 24-0691 -WOPreparation method: SPPS Method A; Lysine Side-Chain Coupling Method B; Cleavage and Purification Method D. Purity: 97.4%; LCMS m / z: Calc’d for C209H321N59O65 4697.36, found 1567.65 [M + 3H]3+. See FIG. 5C and FIG. 6C.Preparation of SBI-404SBI-404 comprises a GLP-1 analog moiety found in dulaglutide.Preparation method: SPPS Method A; Lysine Side-Chain Coupling Method B; Cleavage and Purification Method D. Purity: 95.3%; LCMS m / z Calc’d for C210H319N57O68 4727.33, found 1575.50 [M - 3H]3-. See FIG. 5D and FIG. 6D.Preparation of SBI-405SBI-405 comprises an amylin analog moiety.Preparation method: SPPS Method A; On-resin Disulfide Bond Formation Method C; Cleavage and Purification Method D. Purity: 95.0%; LCMS m / z: Calc’d forCi95H3iiN55O65S24527.22, found 1511.20 [M + 3H]3+. See FIG. 5E and FIG. 6E.MBHB Ref. No.: 24-0691 -WOPreparation of SBI-406SBI-406 comprises an amylin analog moiety.Preparation method: SPPS Method A; On-resin Disulfide Bond Formation Method C; Cleavage and Purification Method D. Purity: 93.9%; LCMS m / z. Calc’d forCi79H2soN54056S24146.02, found 1383.85 [M + 3H]3+. See FIG. 5F and FIG. 6F.Preparation of SB 1-407SBI-407 comprises a GLP-1 / GIP analog moiety.Preparation method: SPPS Method A; Lysine Side-Chain Coupling Method B; Cleavage and Purification Method D. Purity: 94.1%; LCMS m / z Calc’d for C214H325N47O68 4641.34, found 1160.10 [M - 4H]4-. See FIG. 5G and FIG. 6G.MBHB Ref. No.: 24-0691 -WOPreparation of SBI-408SBI-408 comprises a GLP-1 / GCGR agonist moiety.Preparation method: SPPS Method A; Lysine Side-Chain Coupling Method B; Cleavage and Purification Method D. Purity: 94.4%; LCMS m / z: Calc’d for C172H257N43O63 3932.82, found 1312.60 [M + 3H]3+. See FIG. 5H and FIG. 6H.Preparation of SBI-409SBI-409 comprises a GLP-1 / GCGR agonist moiety.Preparation method: SPPS Method A; Lysine Side-Chain Coupling Method B; Cleavage and Purification Method D. Purity: 95.2%; LCMS m / z Calc’d for Ci93H3oiBrN48063 4378.10, found 1461.55 [M + 3H]3+. See FIG. 5I and FIG. 6I.MBHB Ref. No.: 24-0691 -WOPreparation of SB 1-410SBI-410 comprises a GLP-1 / GCGR agonist moiety.Preparation method: SPPS Method A; Lysine Side-Chain Coupling Method B; Cleavage and Purification Method D. Purity: 96.0%; LCMS m / z Calc’d for C190H289N49O61 4233.10, found 1412.90 [M + 3H]3+. See FIG. 5J and FIG. 6J.Preparation of SBI-411SBI-411 comprises an apelin analog moiety.Preparation method: SPPS Method A; Lysine Side-Chain Coupling Method B; Cleavage and Purification Method D. Purity: 95.7%; LCMS m / z: Calc’d for C95H151N25O27S 2106.09, found 1054.60 [M + 2H]2+. See FIG. 5K and FIG. 6K.MBHB Ref. No.: 24-0691 -WOPreparation of SB 1-412Preparation method: SPPS Method A; Cleavage and Purification Method D. Purity: 91.5%; LCMS m / z Calc’d for C124H192IN33O302750.36, found 1376.85 [M + 2H]2+. See Fig. 5L and Fig. 6L.Example 3: Preparation and Testing of Antibody-Peptide Conjugates

[0222] Preparation of antibodv-peptide conjugates from the maleimide-peptides. Anti-CB1 antibody (10 mg / mL) was partially reduced by addition of 3.0 molar equivalents of DTT at pH 8.0, followed by incubation at 37 °C for ~2 h. The reduction reaction mixture was then chilled to ~10 °C and the excess DTT removed via diafiltration. The maleimide-peptide (4.5 eq.) was added. The resulting mixture was incubated at 10 °C for 1 h. After conjugation, excess free cysteine was added to quench the unreacted maleimide-peptide. The reaction mixture was purified and buffer-exchanged into PBS by diafiltration to obtain anti-CB1-peptide conjugates with a DAR of 3.5-4. The molar ratio of peptide substitution was determined using hydrophobic liquid chromatography (HIC). For the preparation of conjugates with a DAR of 1-2, 1.5 eq. of DTT was used for antibody reduction and 2.5 eq. of maleimide-peptide was used for conjugation.

[0223] Preparation of antibodv-peptide conjugates from the bromoacetamido-peptides (SBI-401 and SBI-409). Anti-CB1 antibody (10 mg / mL) was partially reduced by addition of 3.0 molar equivalents of DTT at pH 8.0, followed by incubation at 37 °C for ~2 h. The reduction reaction mixture was then chilled to ~10 °C and the excess DTT removed via diafiltration. The partially reduced anti-CB1 antibody was incubated with 8.0 eq. BrAc-peptide at 25 °C for 4 h. Subsequent purification and characterization are the same as those described in maleimide-peptideMBHB Ref. No.: 24-0691 -WOconjugation above. For the preparation of conjugates with a DAR of 1-2, 1.5 eq. of DTT was used for antibody reduction and 4.0 eq. of BrAc-peptide was used for conjugation.

[0224] Bioconiuqate potency assessment. Antibody-peptide conjugates were dually screened for potency using cell-based functional assessments. First, conjugates were screened for nimacimab potency via cell-based functional assessment using a human CB1 reporter assay kit (Indigo Biosciences, cat. no. IB19001). Then, conjugates were screened for peptide potency via cell-based functional assessment against five obesity-related receptors as shown in Table 5 below.Table 5: Functional assessments for peptide potency

[0225] Results of the functional assessments are shown below.CB1 Inhibition

[0226] Nimacimab retained its potency following peptide conjugation. The results of these potency experiments can be found in FIGS. 7A-J and in Table 6 below. Conjugates were tested at a drug-to-antibody ratio of 2 (P1C4-peptide-D2) and 4 (P1C4-peptide-D4). P1C4 is an alias for nimacimab.Table 6: CB1 inhibition assaysMBHB Ref. No.: 24-0691 -WOPeptide PotencyGLP-1 Receptor Activation: P1C4-401, P1C4-403, P1C4-404

[0227] Conjugates with DAR 2 and 4 were screened for potency (readout: GLP-1 receptor activation) and compared to respective peptide alone, nimacimab, GLP-1 (residues 7-36), and / or tirzepatide. Results are shown in FIGS. 8A-8G, FIGS. 9A-9G, FIGS. 10A-10C, FIGS. 11A-11C, and Table 7 below.Table ?: GLP-1 Receptor Conjugates: Potency AssaysMBHB Ref. No.: 24-0691 -WOAmylin Receptor Activation: P1C4-405, P1C4-406

[0228] Conjugates with DAR 2 and 4 were screened for potency (readout: amylin receptor activation) and compared to respective peptide alone and amylin. Results are shown in FIGS.12A-12B and in Table 8 below.Table 8: Amylin Receptor Conjugates: Potency AssaysGIP Receptor Activation: P1C4-407MBHB Ref. No.: 24-0691 -WO

[0229] Conjugates with DAR 2 and 4 were screened for potency (readout: GIP receptor activation) and compared to respective peptide alone, nimacimab, tirzepatide, and GIP. Results are shown in FIG. 13 and in Table 9 below.Table 9: GIP Receptor Conjugates: Potency AssaysGlucagon Receptor (GCGR) Activation: P1C4-408, P1C4-409, P1C4-410

[0230] Conjugates with DAR 2 and 4 were screened for potency (readout: GCGR activation) and compared to respective peptide alone, nimacimab, retatrutide, and glucagon. Results are shown in FIG. 14, FIGS. 15A-15C, and Table 10 below.Table 10: Glucagon Receptor Conjugates: Potency AssaysApelin Receptor: P1C4-411MBHB Ref. No.: 24-0691 -WO

[0231] Conjugates with DAR 2 and 4 were screened for potency (readout: apelin receptor activation) and compared to respective peptide alone and apelin-13. Results are shown in FIG.16 and in Table 11 below.Table 11 : Apelin Receptor Conjugates: Potency Assays

[0232] In sum, conjugates P1C4-401, P1C4-403, P1C4-404, P1C4-405, P1C4-406, P1C4- 407, P1C4-409, P1C4-410, and P1C4-411 were active at their intended secondary targets (in addition to CB1 inhibition through nimacimab) after conjugation with drug-antibody ratios of 2 and 4. In some cases, increasing the DAR increased potency, whereas in other cases, increasing the DAR decreased potency. Conjugate P1C4-408 was not active after conjugation.

[0233] Having described the invention in detail and by reference to specific embodiments thereof, it will be apparent that modifications and variations are possible without departing from the scope of the invention defined in the appended claims. More specifically, although some aspects of the present invention are identified herein as particularly advantageous, it is contemplated that the present invention is not necessarily limited to these particular aspects of the invention.Table 12: List of SequencesMBHB Ref. No.: 24-0691 -WOMBHB Ref. No.: 24-0691 -WO

Claims

1. MBHB Ref. No.: 24-0691 -WOWhat is claimed is:

1. An isolated peptide, comprising the amino acid sequence:Xaa 1 -Xaa2-Xaa3-Xaa4-Xaa5-Xaa6-Xaa7-Xaa8-Xaa9-Xaa 10-Xaa 11 -Xaa 12-Xaa 13- Xaa 14-Xaa 15-Xaa 16-Xaa 17-Xaa 18-Xaa 19-Xaa20-Xaa21 -Xaa22-Xaa23-Xaa24- Xaa25-Xaa26-Xaa27-Xaa28-Xaa29-Xaa30-Xaa31-Xaa32-Xaa33-Xaa34-Xaa35- Xaa36-Xaa37-Xaa38-Xaa39-Xaa40 (SEQ ID NO: 1);wherein:Xaa1 is His, 3-Mal-PEGo-2o-Lys, 3-Mal-Lys, Tyr, pyroGlu, or 3-Mal-PEGo-2o; Xaa2 is Aib, Gly, Cys, Ser, Arg, or hArg;Xaa3 is Glu, Asn, Gin, Pro, or D-Arg;Xaa4 is Gly, Thr, Arg, or Gin;Xaa5 is Thr, Ala, Leu, or hArg;Xaa6 is Phe, Thr, Ser, or Pro;Xaa7 is Thr, Cys, His, or D-Arg;Xaa8 is Ser, Ala, E-3-Mal-PEGo-2o-Lys, or NMe-Leu;Xaa9 is Asp, Thr, Gly, or Ser;Xaa10 is Vai, Glu, Tyr, £-3-Mal-PEGo-2o-Lys, Pro, or His;Xaa11 is Ser, Arg, Met, or Lys;Xaa12 is Ser, Leu, lie, Glu, Lys, Pro, or Gly;Xaa13 is Tyr, Ala, Aib, Phe, or Oic;Xaa14 is Leu, Glu, NH, or pl-Phe;Xaa15 is Glu, Phe, Asp, Pro, or is absent;Xaa16 is Aib, Gly, Glu, Leu, Lys, Ser, D-Bip, or is absent;Xaa17 is Gin, Arg, lie, Glu, OH, or is absent;Xaa18 is Ala, His, Arg, or is absent;Xaa19 is Ala, Ser, Gin, or is absent;Xaa20 is Lys, Ser, £-3-Mal-PEGo-2o-Lys, Arg, or is absent;Xaa21 is Glu, Asn, Ala, Asp, or is absent;Xaa22 is Phe, Asn, or is absent;Xaa23 is lie, Phe, Vai, or is absent;Xaa24 is Ala, Gly, Gin, Ala, Glu, or is absent;Xaa25 is Trp, Pro, Thr, or is absent;Xaa26 is Leu, lie, or is absent;MBHB Ref. No.: 24-0691 -WOXaa27 is Vai, Leu, lie, Glu, or is absent;Xaa28 is Lys, Pro, Ala, Asn, or is absent;Xaa29 is Gly, Thr, Pro, or is absent;Xaa30 is Gly, Arg, Asn, Thr, e-BrAc-P EG0-20- Lys, PEGO-2o-[£-3-Mal-D-Lys], or is absent; Xaa31 is Gly, (Sar)2.2o-[E-3-Mal-Lys], Vai, Asn, Pro, OH, NH2, or is absent; Xaa32 is (Gly-Gly-Gly-Gly-Ser)3-[£-BrAc-Lys], PEGo-2o-(£-3-Mal-Lys), NH2, (Sar)2.20-[£- 3-Mal-Lys], Gly, Vai, Ser, or is absent;Xaa33 is NH2, Ser, Gly, or is absent;Xaa34 is Asn, Ser, Gly, or is absent;Xaa35 is Thr, Asn, Ala, or is absent;Xaa36 is Pro, Thr, or is absent;Xaa37 is NH2, Pro, or is absent;Xaa38 is NH2, Pro, or is absent;Xaa39 is Ser or is absent; andXaa40 is NH2or is absent.

2. The peptide of claim 1 , wherein Xaa1 is His, Xaa2 is Aib, Xaa3 is Glu, Xaa4 is Gly,Xaa5 is Thr, Xaa6 is Phe, Xaa7 is Thr, Xaa8 is Ser, Xaa9 is Asp, Xaa10 is Vai, Xaa11 is Ser, Xaa12 is Ser, Xaa13 is Tyr, Xaa14 is Leu, Xaa15 is Glu, Xaa16 is Aib, Xaa17 is Gin, Xaa18 is Ala, Xaa19 is Ala, Xaa20 is Lys, Xaa21 is Glu, Xaa22 is Phe, Xaa23 is lie, Xaa24 is Ala, Xaa25 is Trp, Xaa26 is Leu, Xaa27 is Vai, Xaa28 is Lys, Xaa29 is Gly, Xaa30 is Gly, Xaa31 is Gly, Xaa32 is (Gly-Gly-Gly-Gly-Ser)3-[£-BrAc-Lys], Xaa33 is NH2, and Xaa34-Xaa40 are absent relative to SEQ ID NO: 1.

3. The peptide of claim 1, wherein Xaa1 is His, Xaa2 is Gly, Xaa3 is Glu, Xaa4 is Gly,Xaa5 is Thr, Xaa6 is Phe, Xaa7 is Thr, Xaa8 is Ser, Xaa9 is Asp, Xaa10 is Vai, Xaa11 is Ser, Xaa12 is Ser, Xaa13 is Tyr, Xaa14 is Leu, Xaa15 is Glu, Xaa16 is Gly, Xaa17 is Gin, Xaa18 is Ala, Xaa19 is Ala, Xaa20 is Lys, Xaa21 is Glu, Xaa22 is Phe, Xaa23 is lie, Xaa24 is Ala, Xaa25 is Trp, Xaa26 is Leu, Xaa27 is Vai, Xaa28 is Lys, Xaa29 is Gly, Xaa30 is Arg, Xaa31 is (Sar)i6-[£-3-Mal-Lys], Xaa32 is NH2, and Xaa33-Xaa40 are absent relative to SEQ ID NO: 1.

4. The peptide of claim 1, wherein Xaa1 is His, Xaa2 is Gly, Xaa3 is Glu, Xaa4 is Gly,Xaa5 is Thr, Xaa6 is Phe, Xaa7 is Thr, Xaa8 is Ser, Xaa9 is Asp, Xaa10 is Vai, Xaa11MBHB Ref. No.: 24-0691 -WOis Ser, Xaa12 is Ser, Xaa13 is Tyr, Xaa14 is Leu, Xaa15 is Glu, Xaa16 is Glu, Xaa17 is Gin, Xaa18 is Ala, Xaa19 is Ala, Xaa20 is Lys, Xaa21 is Glu, Xaa22 is Phe, Xaa23 is lie, Xaa24 is Ala, Xaa25 is Trp, Xaa26 is Leu, Xaa27 is Vai, Xaa28 is Lys, Xaa29 is Gly, Xaa30 is Gly, Xaa31 is Gly, Xaa32 is (Sar)i6-[c-3-Mal-Lys], Xaa33 is NH2, and Xaa34-Xaa40 are absent relative to SEQ ID NO: 1.

5. The peptide of claim 1, wherein Xaa1 is 3-Mal-PEGs-Lys, Xaa2 is Cys, Xaa3 is Asn,Xaa4 is Thr, Xaa5 is Ala, Xaa6 is Thr, Xaa7 is Cys, Xaa8 is Ala, Xaa9 is Thr, Xaa10 is Gin, Xaa11 is Arg, Xaa12 is Leu, Xaa13 is Ala, Xaa14 is Glu, Xaa15 is Phe, Xaa16 is Leu, Xaa17 is Arg, Xaa18 is His, Xaa19 is Ser, Xaa20 is Ser, Xaa21 is Asn, Xaa22 is Asn, Xaa23 is Phe, Xaa24 is Gly, Xaa25 is Pro, Xaa26 is lie, Xaa27 is Leu, Xaa28 is Pro, Xaa29 is Pro, Xaa30 is Thr, Xaa31 is Asn, Xaa32 is Vai, Xaa33 is Gly, Xaa34 is Ser, Xaa35 is Asn, Xaa36 is Thr, Xaa37 is Pro, Xaa38 is NH2, Xaa39-Xaa40 are absent, wherein the peptide further contains a disulfide bridge, Cys2-Cys7 relative to SEQ ID NO: 1.

6. The peptide of claim 1 , wherein Xaa1 is 6-Mal-Lys, Xaa2 is Cys, Xaa3 is Asn, Xaa4 is Thr, Xaa5 is Ala, Xaa6 is Thr, Xaa7 is Cys, Xaa8 is Ala, Xaa9 is Thr, Xaa10 is Gin, Xaa11 is Arg, Xaa12 is Leu, Xaa13 is Ala, Xaa14 is Glu, Xaa15 is Phe, Xaa16 is Leu, Xaa17 is Arg, Xaa18 is His, Xaa19 is Ser, Xaa20 is Ser, Xaa21 is Asn, Xaa22 is Asn, Xaa23 is Phe, Xaa24 is Gly, Xaa25 is Pro, Xaa26 is lie, Xaa27 is Leu, Xaa28 is Pro, Xaa29 is Pro, Xaa30 is Thr, Xaa31 is Asn, Xaa32 is Vai, Xaa33 is Gly, Xaa34 is Ser, Xaa35 is Asn, Xaa36 is Thr, Xaa37 is Pro, Xaa38 is NH2, Xaa39-Xaa40 are absent, wherein the peptide further contains a disulfide bridge, Cys2-Cys7 relative to SEQ ID NO: 1.

7. The peptide of claim 1, wherein Xaa1 is Tyr, Xaa2 is Aib, Xaa3 is Glu, Xaa4 is Gly,Xaa5 is Thr, Xaa6 is Phe, Xaa7 is Thr, Xaa8 is Ser, Xaa9 is Asp, Xaa10 is Tyr, Xaa11 is Ser, Xaa12 is lie, Xaa13 is Aib, Xaa14 is Leu, Xaa15 is Asp, Xaa16 is Lys, Xaa17 is lie, Xaa18 is Ala, Xaa19 is Gin, Xaa20 is c-3-Mal-PEGs-Lys, Xaa21 is Ala, Xaa22 is Phe, Xaa23 is Vai, Xaa24 is Gin, Xaa25 is Trp, Xaa26 is Leu, Xaa27 is lie, Xaa28 is Ala, Xaa29 is Gly, Xaa30 is Gly, Xaa31 is Pro, Xaa32 is Ser, Xaa33 is Ser, Xaa34 is Gly, Xaa35 is Ala, Xaa36 is Pro, Xaa37 is Pro, Xaa38 is Pro, Xaa39 is Ser, Xaa40 is NH2relative to SEQ ID NO: 1.MBHB Ref. No.: 24-0691 -WO8. The peptide of claim 1, wherein Xaa1 is His, Xaa2 is Ser, Xaa3 is Gin, Xaa4 is Gly,Xaa5 is Thr, Xaa6 is Phe, Xaa7 is Thr, Xaa8 is Ser, Xaa9 is Asp, Xaa10 is s-3-Mal- PEGs-Lys, Xaa11 is Ser, Xaa12 is Glu, Xaa13 is Tyr, Xaa14 is Leu, Xaa15 is Asp, Xaa16 is Ser, Xaa17 is Glu, Xaa18 is Arg, Xaa19 is Ala, Xaa20 is Arg, Xaa21 is Asp, Xaa22 is Phe, Xaa23 is Vai, Xaa24 is Ala, Xaa25 is Trp, Xaa26 is Leu, Xaa27 is Glu, Xaa28 is Ala, Xaa29 is Gly, Xaa30 is Gly, Xaa31 is OH, and Xaa32-Xaa40 are absent relative to SEQ ID NO: 1.

9. The peptide of claim 1, wherein Xaa1 is His, Xaa2 is Aib, Xaa3 is Gin, Xaa4 is Gly,Xaa5 is Thr, Xaa6 is Phe, Xaa7 is Thr, Xaa8 is Ser, Xaa9 is Asp, Xaa10 is Tyr, Xaa11 is Ser, Xaa12 is Lys, Xaa13 is Tyr, Xaa14 is Leu, Xaa15 is Asp, Xaa16 is Glu, Xaa17 is Arg, Xaa18 is Arg, Xaa19 is Ala, Xaa20 is Arg, Xaa21 is Asp, Xaa22 is Phe, Xaa23 is Vai, Xaa24 is Glu, Xaa25 is Trp, Xaa26 is Leu, Xaa27 is Leu, Xaa28 is Asn, Xaa29 is Thr, Xaa30 is £-BrAc-PEGi2-Lys, Xaa31 is NH2, and Xaa32-Xaa40 are absent relative to SEQ ID NO: 1.

10. The peptide of claim 1, wherein Xaa1 is His, Xaa2 is Aib, Xaa3 is Gin, Xaa4 is Gly,Xaa5 is Thr, Xaa6 is Phe, Xaa7 is Thr, Xaa8 is Ser, Xaa9 is Asp, Xaa10 is Tyr, Xaa11 is Ser, Xaa12 is Lys, Xaa13 is Tyr, Xaa14 is Leu, Xaa15 is Asp, Xaa16 is Glu, Xaa17 is Arg, Xaa18 is Arg, Xaa19 is Ala, Xaa20 is Arg, Xaa21 is Asp, Xaa22 is Phe, Xaa23 is Vai, Xaa24 is Glu, Xaa25 is Trp, Xaa26 is Leu, Xaa27 is Leu, Xaa28 is Asn, Xaa29 is Thr, Xaa30 is PEGs-[E-3-Mal-D-Lys], Xaa31 is NH2, and Xaa32-Xaa40 are absent relative to SEQ ID NO: 1.

11. The peptide of claim 1, wherein Xaa1 is pyroGlu, Xaa2 is Arg, Xaa3 is Pro, Xaa4 is Arg, Xaa5 is Leu, Xaa6 is Ser, Xaa7 is His, Xaa8 is e-3-Mal-PEGs-Lys, Xaa9 is Gly, Xaa10 is Pro, Xaa11 is Met, Xaa12 is Pro, Xaa13 is Phe, Xaa14 NH, and Xaa15- Xaa40 are absent relative to SEQ ID NO: 1.

12. The peptide of claim 1 , wherein the peptide has the amino acid sequence:(a) His-Aib-Glu-Gly-Thr-Phe-Thr-Ser-Asp-Val-Ser-Ser-Tyr-Leu-Glu-Aib-GIn- Ala-Ala-Lys-Glu-Phe-lle-Ala-Trp-Leu-Val-Lys-Gly-Gly-Gly-(Gly-Gly-Gly- Gly-Ser)3-[£-BrAc-Lys]-NH2(SEQ ID NO: 2), wherein [£-BrAc-Lys]-NH2has the following structure:MBHB Ref. No.: 24-0691 -WO(b) His-Gly-Glu-Gly-Thr-Phe-Thr-Ser-Asp-Val-Ser-Ser-Tyr-Leu-Glu-Gly-GIn- Ala-Ala-Lys-Glu-Phe-lle-Ala-Trp-Leu-Val-Lys-Gly-Arg-(Sar)i6-[£-3-Mal- Lys]-NH2 (SEQ ID NO: 4), wherein -[E-3-Mal-Lys]-NH2 has the following structure:(c) His-Gly-Glu-Gly-Thr-Phe-Thr-Ser-Asp-Val-Ser-Ser-Tyr-Leu-Glu-Glu-GIn- Ala-Ala-Lys-Glu-Phe-lle-Ala-Trp-Leu-Val-Lys-Gly-Gly-Gly-(Sar)i6-[£-3- Mal-Lys]-NH2 (SEQ ID NO: 5), wherein [e-3-Mal-Lys]-NH2 has the following structure:(d) [3-Mal-PEG8-Lys]-Cys-Asn-Thr-Ala-Thr-Cys-Ala-Thr-Gln-Arg-Leu-Ala- Glu-Phe-Leu-Arg-His-Ser-Ser-Asn-Asn-Phe-Gly-Pro-lle-Leu-Pro-Pro-Thr- Asn-Val-Gly-Ser-Asn-Thr-Pro-NH2(disulfide bridge, Cys2-Cys7) (SEQ ID NO: 6), wherein [3-Mal-PEG8-Lys] has the following structure:MBHB Ref. No.: 24-0691 -WO(e) [6-Mal-l_ys]-Cys-Asn-Thr-Ala-Thr-Cys-Ala-Thr-Gln-Arg-Leu-Ala-Glu-Phe- Leu-Arg-His-Ser-Ser-Asn-Asn-Phe-Gly-Pro-lle-Leu-Pro-Pro-Thr-Asn-Val- Gly-Ser-Asn-Thr-Pro-NH2(disulfide bridge, Cys2-Cys7) (SEQ ID NO: 7), wherein [6-Mal-Lys]- has the following structure:(f) Tyr-Aib-Glu-Gly-Thr-Phe-Thr-Ser-Asp-Tyr-Ser-lle-Aib-Leu-Asp-Lys-lle- Ala-Gln-[£-3-Mal-PEG8-Lys]-Ala-Phe-Val-Gln-Trp-Leu-lle-Ala-Gly-Gly-Pro- Ser-Ser-Gly-Ala-Pro-Pro-Pro-Ser-NH2(SEQ ID NO: 8), wherein -[£-3-Mal- PEGs-Lys] has the following structure:(g) His-Ser-Gln-Gly-Thr-Phe-Thr-Ser-Asp-[£-3-Mal-PEG8-Lys]-Ser-Glu-Tyr- Leu-Asp-Ser-Glu-Arg-Ala-Arg-Asp-Phe-Val-Ala-Trp-Leu-Glu-Ala-Gly-Gly- OH (SEQ ID NO: 9), wherein -[£-3-Mal-PEG8-Lys] has the following structure:(h) His-Aib-GIn-Gly-Thr-Phe-Thr-Ser-Asp-Tyr-Ser-Lys-Tyr-Leu-Asp-Glu-Arg- Arg-Ala-Arg-Asp-Phe-Val-Glu-Trp-Leu-Leu-Asn-Thr-[£-BrAc-PEGi2-Lys]-MBHB Ref. No.: 24-0691 -WONH2(SEQ ID NO: 10), wherein [£-BrAc-PEGi2-Lys]-NH2 has the following structure:(i) His-Aib-GIn-Gly-Thr-Phe-Thr-Ser-Asp-Tyr-Ser-Lys-Tyr-Leu-Asp-Glu-Arg- Arg-Ala-Arg-Asp-Phe-Val-Glu-Trp-Leu-Leu-Asn-Thr-PEG8-[£-3-Mal-D- Lys]-NH2(SEQ ID NO: 11), wherein -[PEG8-£-3-Mal-D-Lys]-NH2has the following structure:(j) [pyroGlu]-Arg-Pro-Arg-Leu-Ser-His-[£-3-Mal-PEG8-Lys]-Gly-Pro-Met-Pro- Phe-NH2(SEQ ID NO: 12), wherein [e-3-Mal-PEGs-Lys] has the following structure:

13. A pharmaceutical composition, comprising the peptide of any one of claims 1-12 and a pharmaceutically acceptable excipient, carrier, or diluent.

14. A kit comprising the peptide of any one of claims 1-12 or the pharmaceutical composition of claim 13.

15. A conjugate, comprising:(a) at least one of the peptides of any one of claims 1-12; and(b) an isolated antibody or an antigen binding fragment thereof that specifically binds to human cannabinoid 1 (CB1) receptor;MBHB Ref. No.: 24-0691 -WOwherein isolated antibody or antigen binding fragment thereof comprises: (i) a heavy chain variable (VH) region comprising the following complementarity determining regions (CDRs):HCDR1 having the amino acid sequence of SEQ ID NO: 26; HCDR2 having the amino acid sequence of SEQ ID NO: 27; HCDR3 having the amino acid sequence of SEQ ID NO: 28; and(ii) a light chain variable (VL) region comprising the following CDRs:LCDR1 having the amino acid sequence of SEQ ID NO: 29; LCDR2 having the amino acid sequence of SEQ ID NO: 30; LCDR3 having the amino acid sequence of SEQ ID NO: 31; or or a pharmaceutically acceptable salt, solvate, stereoisomer, or tautomer thereof.

16. The conjugate of claim 15, wherein the VH region of the isolated antibody or the antigen binding fragment thereof comprises an amino acid sequence that is 90% identical to SEQ ID NO: 32.

17. The conjugate of claim 15, wherein the VL region of the isolated antibody or the antigen binding fragment thereof comprises an amino acid sequence that is 90% identical to SEQ ID NO: 33.

18. The conjugate of claim 15, wherein the isolated antibody or the antigen binding f ragment thereof comprises:(a) a heavy chain comprising an amino acid sequence at least 90% identical to SEQ ID NO: 32; and(b) a light chain variable region comprising an amino acid sequence that is at least 90% identical to SEQ ID NO: 33.

19. The conjugate of claim 15, wherein the conjugate has a drug-antibody ratio of the at least one of the peptides of any one of claims 1 -4 to the isolated antibody or the antigen binding fragment thereof, wherein the drug-antibody ratio is greater than 1.

20. The conjugate of claim 19, wherein the drug-antibody ratio is about 2.MBHB Ref. No.: 24-0691 -WO21. The conjugate of claim 19, wherein the drug-antibody ratio is about 4.

22. The conjugate of any one of claims 15-21, wherein the at least one of the peptides is linked to the isolated antibody or the antigen binding fragment thereof via cysteine- based bioconjugation.

23. The conjugate of any one of claims 15-21, wherein the at least one of the peptides is linked site-specifically to the isolated antibody or the antigen binding fragment thereof.

24. A pharmaceutical composition, comprising the conjugate of any one of claims 15-23 and a pharmaceutically acceptable excipient, carrier, or diluent.

25. A kit comprising the conjugate of any one of claims 15-21 or the pharmaceutical composition of claim 24.

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