Conjugates containing phosphorus(V) and a drug moiety

Abstract

The present invention relates to a conjugate having formula (I), in which a receptor binding molecule (RBM) is linked to a drug moiety (D). The present invention also relates to intermediates for producing same, processes for preparing same, pharmaceutical compositions containing same, and uses thereof. TIFF2024540691000549.tif35128

Description

[Technical field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application claims the benefit of priority to European Patent Application No. 21207195.5, filed November 9, 2021, the contents of which are incorporated herein by reference in their entirety for all purposes.

[0002] Technical Field The present invention relates to conjugates of receptor binding molecules and drug moieties, intermediates for producing same, processes for preparing same, pharmaceutical compositions containing same, and uses thereof. [Background technology]

[0003] background Brentuximab vedotin (trade name ADCETRIS®) is an antibody-drug conjugate approved for medical use in 2011. Brentuximab vedotin consists of a tumor-targeting chimeric IgG1 antibody component, brentuximab, and a linker-payload component that contains monomethyl auristatin E, a payload moiety that induces apoptosis upon intracellular delivery and release.

[0004] However, although brentuximab vedotin is an approved and marketed ADC, it still has drawbacks. For example, brentuximab vedotin is known to have a certain limit on the number of drug molecules that can be attached to the antibody. For example, see Hamblett et al., "Effects of Drug Loading on the Antitumor Activity of a Monoclonal Antibody Drug Conjugate", Clinical Cancer Research vol. 10, pp.7063 to 7070, October 15, 2004, https: / / doi.org / 10.1158 / 1078-0432.CCR-04-0789. Development includes, for example, changing the linker that connects the antibody to the payload and introducing polyethylene glycol substituents.For example, see Lyon et al., "Reducing hydrophobicity of homogeneous antibody-drug conjugates improves pharmacokinetics and therapeutic index", Nature Biotechnology volume 33, pages 733-735 (2015), doi: 10.1038 / nbt.3212 (Non-Patent Document 2); WO 2015 / 057699 (Patent Document 1); Burke et al., "Optimization of a PEGylated Glucuronide-Monomethylauristatin E Linker for Antibody-Drug Conjugates", Molecular Cancer Therapeutics 2017, 16(1), 116-123, doi: 10.1158 / 1535-7163.MCT-16-0343 (Non-Patent Document 3); and Simmons et al., "Reducing the antigen-independent toxicity of antibody-drug conjugates by See, “minimizing their non-specific clearance through PEGylation”, Toxicology and Applied Pharmacology 2020, 392:114932, doi: 10.1016 / j.taap.2020.114932 (Non-Patent Document 4).

[0005] Thus, there is a continuing need for additional conjugates that have good properties for pharmaceutical applications. [Prior art documents] [Patent documents]

[0006] [Patent Document 1] WO 2015 / 057699 [Non-patent literature]

[0007] [Non-Patent Document 1] Hamblett et al., “Effects of Drug Loading on the Antitumor Activity of a Monoclonal Antibody Drug Conjugate”, Clinical Cancer Research vol. 10, pp.7063 to 7070, October 15, 2004, https: / / doi.org / 10.1158 / 1078-0432.CCR-04-0789 [Non-Patent Document 2] Lyon et al., “Reducing hydrophobicity of homogeneous antibody-drug conjugates improves pharmacokinetics and therapeutic index”, Nature Biotechnology volume 33, pages 733-735 (2015), doi: 10.1038 / nbt.3212 [Non-Patent Document 3] Burke et al., “Optimization of a PEGylated Glucuronide-Monomethylauristatin E Linker for Antibody-Drug Conjugates”, Molecular Cancer Therapeutics 2017, 16(1), 116-123, doi: 10.1158 / 1535-7163.MCT-16-0343 [Non-Patent Document 4] Simmons et al., “Reducing the antigen-independent toxicity of antibody-drug conjugates by minimizing their non-specific clearance through PEGylation”, Toxicology and Applied Pharmacology 2020, 392:114932, doi: 10.1016 / j.taap.2020.114932 Summary of the Invention

[0008] overview This need is addressed by the subject matter as defined in the claims and in the embodiments described herein.

[0009] Thus, the present invention provides a conjugate having formula (I): TIFF2024540691000002.tif35128, or a pharma- ceutically acceptable salt or solvate thereof; During the ceremony: RBM is receptor binding molecule; TIFF2024540691000003.tif6128 is a double bond; or TIFF2024540691000004.tif6128 is a single bond; V is not present if TIFF2024540691000005.tif6128 is a double bond; or V is If TIFF2024540691000006.tif6128 is a single bond, it is H or (C 1 -C 8 ) alkyl; X is R if TIFF2024540691000007.tif6128 is a double bond 3 -C; or X is If TIFF2024540691000008.tif6128 is a single bond, TIFF2024540691000009.tif9128; Y is NR 5 , S, O, or CR 6 R 7 and; R 1 is a first polyalkylene glycol unit R comprising at least three alkylene glycol subunits; F and; R 3 is H; or an optionally substituted aliphatic or an optionally substituted aromatic residue; R 4 is H; or an optionally substituted aliphatic or an optionally substituted aromatic residue; R 5 is H; or an optionally substituted aliphatic or an optionally substituted aromatic residue; R 6 is H; or an optionally substituted aliphatic or an optionally substituted aromatic residue; R 7 is H; or an optionally substituted aliphatic or an optionally substituted aromatic residue; L is a linker; D is a drug moiety; m is an integer ranging from 1 to 10; and n is an integer ranging from 1 to 20.

[0010] The present invention also relates to a compound having the formula (II): TIFF2024540691000010.tif29128, or a pharma- ceutically acceptable salt or solvate thereof; During the ceremony: TIFF2024540691000011.tif6128 is a triple bond; or TIFF2024540691000012.tif6128 is a double bond; V is absent if TIFF2024540691000013.tif6128 is a triple bond; or V is If TIFF2024540691000014.tif6128 is a double bond, H or (C 1 -C 8 ) alkyl; X is R if TIFF2024540691000015.tif6128 is a triple bond 3 -C: or X is If TIFF2024540691000016.tif6128 is a double bond, TIFF2024540691000017.tif9128; Y is NR 5 , S, O, or CR 6 R 7 and; R 1 is a first polyalkylene glycol unit comprising at least three alkylene glycol subunits; R 3 is H; or an optionally substituted aliphatic or an optionally substituted aromatic residue; R 4 is H; or an optionally substituted aliphatic or an optionally substituted aromatic residue; R 5 is H; or an optionally substituted aliphatic or an optionally substituted aromatic residue; R 6 is H; or an optionally substituted aliphatic or an optionally substituted aromatic residue; R 7 is H; or an optionally substituted aliphatic or an optionally substituted aromatic residue; L is a linker; D is a drug moiety; and m is an integer ranging from 1 to 10.

[0011] The present invention also relates to a method for preparing a conjugate of formula (I), the method comprising the steps of: Compounds of formula (II): TIFF2024540691000018.tif29128, or a pharma- ceutically acceptable salt or solvate thereof, During the ceremony: TIFF2024540691000019.tif6128 is a triple bond; or TIFF2024540691000020.tif6128 is a double bond; V is absent if TIFF2024540691000021.tif6128 is a triple bond; or V is If TIFF2024540691000022.tif6128 is a double bond, H or (C 1 -C 8 ) alkyl; X is R if TIFF2024540691000023.tif6128 is a triple bond 3 -C: or X is If TIFF2024540691000024.tif6128 is a double bond, TIFF2024540691000025.tif9128; Y is NR 5 , S, O, or CR 6 R 7 and; R 1 is a first polyalkylene glycol unit R comprising at least three alkylene glycol subunits; F and; R 3 is H; or an optionally substituted aliphatic or an optionally substituted aromatic residue; R 4 is H; or an optionally substituted aliphatic or an optionally substituted aromatic residue; R 5is H; or an optionally substituted aliphatic or an optionally substituted aromatic residue; R 6 is H; or an optionally substituted aliphatic or an optionally substituted aromatic residue; R 7 is H; or an optionally substituted aliphatic or an optionally substituted aromatic residue; L is a linker; D is a drug moiety; and m is an integer ranging from 1 to 10; A compound of formula (II) or a pharma- ceutically acceptable salt or solvate thereof, A thiol-containing molecule of formula (III): TIFF2024540691000026.tif9128, wherein RBM is a receptor binding molecule; and n is an integer ranging from 1 to 20; a thiol-containing molecule of formula (III) By reacting Compounds of formula (I): TIFF2024540691000027.tif35128, or a pharma- ceutically acceptable salt or solvate thereof, During the ceremony: TIFF2024540691000028.tif6128 is a compound of formula (II) TIFF2024540691000029.tif6128 is a triple bond if it is a double bond; or TIFF2024540691000030.tif6128 is a compound of formula (II) TIFF2024540691000031.tif6128 is a double bond, then it is a single bond; V is not present if TIFF2024540691000032.tif6128 is a double bond; or V is If TIFF2024540691000033.tif6128 is a single bond, H or (C 1 -C 8 ) alkyl; X is R if TIFF2024540691000034.tif6128 is a double bond 3 -C; or X is If TIFF2024540691000035.tif6128 is a single bond, TIFF2024540691000036.tif9128; Y is NR 5 , S, O, or CR 6 R 7 and; R 1 is a first polyalkylene glycol unit R comprising at least three alkylene glycol subunits; F and; R 3 is H; or an optionally substituted aliphatic or an optionally substituted aromatic residue; R 4 is H; or an optionally substituted aliphatic or an optionally substituted aromatic residue; R 5 is H; or an optionally substituted aliphatic or an optionally substituted aromatic residue; R 6 is H; or an optionally substituted aliphatic or an optionally substituted aromatic residue; R 7 is H; or an optionally substituted aliphatic or an optionally substituted aromatic residue; L is a linker; D is a drug moiety; m is an integer ranging from 1 to 10; and n is an integer ranging from 1 to 20; A compound of formula (I) or a pharma- ceutically acceptable salt or solvate thereof The method includes the step of generating

[0012] The present invention also relates to a conjugate of formula (I) obtainable or obtainable by the process of the present invention.

[0013] The present invention also relates to a pharmaceutical composition comprising a conjugate of the present invention.

[0014] The present invention also relates to a conjugate of the invention for use in a method of treating a disease. The disease may be cancer.

[0015] The present invention also relates to a pharmaceutical composition of the present invention for use in a method of treating a disease. The disease may be cancer. [Brief description of the drawings]

[0016] [Figure 1] 1 shows an analytical HPLC chromatogram of the compound 4-azido-2-(dodecaethyleneglycol)methyl benzoate. The horizontal axis depicts retention time in minutes. [Diagram 2] 1 shows an analytical HPLC chromatogram of the compound 4-azido-2-(dodecaethyleneglycol)methyl benzoate. The horizontal axis depicts retention time in minutes. [Diagram 3] Figure 2 shows an analytical HPLC chromatogram of compound P5(PEG12)-COOH. [Figure 4] 1 shows an analytical HPLC chromatogram of compound P5(PEG24)-OSu. The horizontal axis depicts retention time in minutes. [Diagram 5] 1 shows an analytical HPLC chromatogram of compound P5(PEG12,PEG24)-COOH. The horizontal axis depicts retention time in minutes. [Figure 6] 1 shows an analytical HPLC chromatogram of compound P5(PEG24,PEG24)-COOH. The horizontal axis depicts retention time in minutes. [Figure 7] 1 shows an analytical HPLC chromatogram of compound NH2-VC-PAB-MMAE TFA salt. The horizontal axis depicts retention time in minutes. [Figure 8]1 shows an analytical HPLC chromatogram of compound P5 (PEG12)-VC-PAB-MMAF. The horizontal axis depicts retention time in minutes. [Figure 9] 1 shows an analytical HPLC chromatogram of compound P5 (PEG12)-VC-PAB-MMAE. The horizontal axis depicts retention time in minutes. [Figure 10] Figure 2 shows an analytical HPLC chromatogram of compound P5 (PEG24)-VC-PAB-MMAE. [Figure 11] 1 shows an analytical HPLC chromatogram of compound P5 (PEG12,PEG24)-VC-PAB-MMAE. The horizontal axis depicts retention time in minutes. [Figure 12] 1 shows an analytical HPLC chromatogram of compound P5 (PEG24,PEG24)-VC-PAB-MMAE. The horizontal axis depicts retention time in minutes. [Figure 13] Figure 1 shows an analytical SEC chromatogram of Trastuzumab, where SEC stands for size exclusion chromatography. [Figure 14] 1 shows an analytical HIC chromatogram of trastuzumab, where HIC stands for hydrophobic interaction chromatography. [Figure 15] 1 shows an analytical SEC chromatogram of brentuximab. [Figure 16] 1 shows an analytical HIC chromatogram of brentuximab. [Figure 17] Analytical SEC chromatogram of Brentuximab-P5(PEG12)-VC-PAB-MMAE (DAR8). [Figure 18] Analytical HIC chromatogram of brentuximab-P5(PEG12)-VC-PAB-MMAE (DAR8). [Figure 19] Analytical SEC chromatogram of Brentuximab-P5(PEG12)-VC-PAB-MMAE (DAR4). [Figure 20] 4 shows analytical SEC chromatogram of brentuximab-P5(PEG24)-VC-PAB-MMAE(DAR8). [Figure 21]Analytical HIC chromatogram of brentuximab-P5(PEG24)-VC-PAB-MMAE (DAR8) is shown. [Figure 22] Analytical SEC chromatogram of brentuximab-P5(PEG12, PEG24)-VC-PAB-MMAE (DAR8). [Diagram 23] Analytical HIC chromatogram of brentuximab-P5(PEG12, PEG24)-VC-PAB-MMAE (DAR8) is shown. [Figure 24] Analytical SEC chromatogram of brentuximab-P5(PEG24, PEG24)-VC-PAB-MMAE (DAR8). [Diagram 25] Analytical HIC chromatogram of brentuximab-P5(PEG24, PEG24)-VC-PAB-MMAE (DAR8) is shown. [Figure 26] Analytical HIC chromatogram of brentuximab-P5(PEG12)-VC-PAB-MMAF(DAR8) is shown. [Figure 27] Analytical SEC chromatogram of Trastuzumab-P5(PEG12)-VC-PAB-MMAE (DAR8). [Figure 28] Analytical HIC chromatogram of Trastuzumab-P5(PEG12)-VC-PAB-MMAE(DAR8). [Figure 29] FIG. 1 shows analytical SEC chromatogram of Trastuzumab-P5(PEG12)-VC-PAB-MMAF(DAR8). [Diagram 30] Analytical HIC chromatogram of Trastuzumab-P5(PEG12)-VC-PAB-MMAF(DAR8) is shown. [Diagram 31]Screening experiments to identify optimal conditions for antibody modification with PEGylated phosphonamidates. Drug-antibody ratios (DARs) were measured by MS. Left graph: 10 equivalents of P5(PEG12)-VC-PAB-MMAE were used in the conditions described above, and the TCEP equivalents were varied. The maximum degree of modification was reached with 8 equivalents of TCEP. Right graph: Those 8 equivalents were carried over to a second experiment, where the P5(PEG12)-VC-PAB-MMAE equivalents were further increased to achieve a maximum DAR of 8. The optimal conditions to achieve a DAR of 8 were identified as 8 equivalents of TCEP and 12 equivalents of P5(PEG12)-VC-PAB-MMAE relative to the antibody (=only 1.5 equivalents per Cys). [Diagram 32] FIG. 1 shows hydrophobic interaction chromatography of brentuximab conjugated to P5(PEG2)-, P5(PEG12)- and P5(PEG24)-VC-PAB-MMAE in a head-to-head comparison with commercially available ADCETRIS (Brentuximab-maleimidocapryl-VC-PAB-MMAE, DAR4av, black). [Diagram 33] Hydrophobic interaction chromatography of brentuximab conjugated to P5(PEG24,PEG12)-, P5(PEG24,PEG24)- and P5(PEG24)-VC-PAB-MMAE in a head-to-head comparison with commercially available ADCETRIS (Brentuximab-Maleimidocapryl-VC-PAB-MMAE, DAR4av, black). #No additional peaks identified. [Diagram 34] Analytical size exclusion chromatography (SEC) (left) and HIC (right) of DAR8 Brentuximab-P5(PEG12)-VC-PAB-MMAE after storage for several weeks. No aggregates are observed in SEC and no drug loss is observed in HIC. [Diagram 35]In vitro cytoxicity of brentuximab (anti-CD30) ADCs in antigen-positive (Karpas 299, left) and antigen-negative (HL-60, right) cell lines. Comparison of three different DAR8 ADCs (PEG2 vs. PEG12 vs. PEG24) differing only in the length of the PEG substituents against unmodified brentuximab. [Diagram 36] In vitro cytotoxicity of brentuximab (anti-CD30) ADC in antigen positive (Karpas 299, left) and antigen negative (HL-60, right) cell lines. Comparison of brentuximab-P5(PEG24)-vc-PAB-MMAE (DAR8) versus commercial Adcetris (DAR4). [Figure 37] In vitro cytotoxicity of brentuximab (anti-CD30) ADCs in antigen-positive (Karpas 299, left) and antigen-negative (HL-60, right) cell lines. Comparison of brentuximab-P5(PEG24)-vc-PAB-MMAE modified with 4 (DAR4) or 8 (DAR8) linker payload molecules per antibody. [Figure 38] In vitro cytotoxicity of brentuximab (anti-CD30) ADC in antigen-positive (Karpas 299, left) and antigen-negative (HL-60, right) cell lines. Comparison of brentuximab-P5(PEG12)-vc-PAB-MMAE (DAR8) with the same construct carrying the MMAF payload is shown. [Figure 39] In vitro cytotoxicity of trastuzumab (anti-Her2) ADC in antigen-positive (SKBR3, left) and antigen-negative (MDAMB, right) cell lines. Comparison of trastuzumab-P5(PEG12)-vc-PAB-MMAF(DAR8) versus unmodified trastuzumab. [Diagram 40]Evaluation of bystander effect depending on the differently PEGylated brentuximab-P5-VC-PAB-MMAE constructs. Top: In vitro cytotoxicity of brentuximab (anti-CD30) ADC in two antigen-positive cell lines (Karpas 299, left, and L-540, right) and an antigen-negative cell line (HL-60, bottom left). To evaluate bystander killing, the supernatant after incubation of L-540 with ADC was transferred to HL-60 (HL60, bottom right). [Diagram 41] Figure 1 shows the in vivo evaluation of brentuximab-(PEG12)-VC-PAB-MMAE (DAR8 and DAR4), Adcetris (DAR4), and untreated control in a Karpas299-based tumor xenograft model in SCID mice with 10 animals per group. Mice were treated with 0.5 mg / kg of constructs every 4 days for 4 times. The graph on the left shows the mean tumor volume of all 10 mice per group. The last observation point of sacrificed animals was carried forward (LOCF). The graph on the right shows the Kaplan-Meier plot of survival in each group. [Diagram 42] Figure 1 shows quantification of total antibody in the circulation following treatment of female Spraque-Dawley rats with either brentuximab-P5(PEG24)-VC-PAB-MMAE or Adcetris by ELISA. [Diagram 43]A) Conjugation of P5(PEG12)-VC-PAB-SB743921 and P5(PEG24)-VA-PAB-SB743921 to trastuzumab; B) Conjugation efficiency estimated by mass spectrometry (MS). Drug-antibody ratios were calculated from the MS intensities of modified and unmodified heavy and light chain species; note that in this example, only little or slow conjugation was observed when using short PEG2 residues (i.e., PEG residues containing 2 PEG units), while more efficient conjugation reactions were achieved when using PEG12 (i.e., PEG residues containing 12 PEG units), and even more efficient conjugation reactions were achieved when using PEG24 (i.e., PEG residues containing 24 PEG units), as shown by the higher drug-antibody ratio (DAR). C) Exemplary MS spectrum of conjugation of P5(PEG24)-VA-PAB-SB743921 to trastuzumab. The mass spectrum shows the unmodified light chain (23438 Da), the modified light chain (25439 Da), the unmodified heavy chain (49149 Da) and the triply modified heavy chain (55452 Da). [Diagram 44] Showing efficacy of ADC trastuzumab-P5(PEG24)-VA-PAB-SB743921 in target negative cell line (L-540) and several Her2+ cell lines. Although trastuzumab-P5(PEG24)-VA-PAB-SB743921 only shows efficacy against non-target L-540 at the highest concentration tested, it was able to show much better efficacy against all target positive cell lines tested. [Diagram 45] A) Conjugation of P5(PEG12)-VC-PAB-emetine to trastuzumab; B) Conjugation efficiency estimated by mass spectrometry (MS). Drug-antibody ratios were calculated from the MS intensities of modified and unmodified heavy and light chain species; C) Exemplary HIC spectrum of conjugation of 16 equivalents of P5(PEG12)-VC-PAB-emetine to trastuzumab, resulting in a DAR 8.0 ADC. No unconjugated trastuzumab was observed at the known retention time of the unmodified antibody (approximately 8-9 min). [Figure 46] Normalized HIC chromatograms of trastuzumab-P5(PEG12)-VC-PAB-emetineDAR8 and Adcetris (brentuximab vedotin) are shown. [Figure 47] A) Conjugation of P5(PEG12)-VC-PAB-AT7519 to trastuzumab; B) Analysis of purified DAR8 ADC by size exclusion chromatography (SEC) and hydrophobic interaction chromatography (HIC); C) MS analysis of conjugation of 16 equivalents of P5(PEG12)-VC-PAB-AT7519 to trastuzumab resulting in a DAR 8.0 ADC. No unconjugated trastuzumab was observed. [Figure 48] A) Conjugation of P5(PEG24)-VA-PAB-panobinostat to trastuzumab; B) Analysis of purified DAR8 ADC by hydrophobic interaction chromatography (HIC). [Figure 49] A) Conjugation of P5(PEG12)-GlcA-AT7519 to trastuzumab; B) Conjugation efficiency estimated by MS depending on the equivalent amount of P5(PEG12)-GlcA-AT7519. Reactions were performed as described herein with 8 equivalents of TCEP. Drug-antibody ratios were calculated from the MS intensities of modified and unmodified heavy and light chain species; C) An exemplary MS spectrum of conjugation of 6 equivalents of P5(PEG12)-GlcA-AT7519 to trastuzumab resulting in a DAR 3.9 ADC is shown. [Figure 50] Normalized HIC chromatograms of trastuzumab-P5(PEG12)-GlcA-AT7519 and Adcetris (brentuximab vedotin); and SEC chromatogram of trastuzumab-P5(PEG12)-GlcA-AT7519. [Figure 51]A) Conjugation of P5(PEG12)-GlcA-MMAE to Brentuximab; B) Efficacy against target negative cell line (HL-60, bottom) and target positive cell line (Karpas299, top); Brentuximab-P5(PEG12)-GlcA-MMAE shows no effect against target negative cell line (HL-60) but can show much better efficacy against target positive cell line Karpas299. [Figure 52] A) Conjugation of P5(PEG12)-GlcA-SB743921 to trastuzumab; B) Conjugation efficiency estimated by mass spectrometry (MS) depending on the equivalents of P5(PEG12)-GlcA-SB743921. Reactions were performed as described herein with 8 equivalents of TCEP. Drug-antibody ratios were calculated from the MS intensities of modified and unmodified heavy and light chain species. C) Analysis of purified DAR8 ADC by size exclusion chromatography (SEC) and hydrophobic interaction chromatography (HIC) are shown. [Figure 53] The efficacy test results of trastuzumab-P5(PEG12)GlcA-SB743921 in target-negative cell line (MDA-MB468) and target-positive cell line (SKBR3) are shown. Trastuzumab-P5(PEG12)-GlcA-SB743921 shows no effect on the target-negative cell line (MDA-MB468), but shows much better efficacy on SKBR-3. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0017] Detailed Description The present invention is described in detail below and is further illustrated by the accompanying examples and figures.

[0018] definition Unless otherwise indicated, the term "alkyl," by itself or as part of another term, generally refers to a substituted or unsubstituted straight or branched chain saturated hydrocarbon having the indicated number of carbon atoms; for example, "-(C 1 -C 8) alkyl" or "-(C 1 -C 10 "C )alkyl" refers to an alkyl group having 1 to 8 or 1 to 10 carbon atoms, respectively. If the number of carbon atoms is not specified, the alkyl group may have 1 to 8 carbon atoms. Representative straight chain -(C 1 -C 8 )Alkyl groups include, but are not limited to, -methyl, -ethyl, -n-propyl, -n-butyl, -n-pentyl, -n-hexyl, -n-heptyl, and -n-octyl; branched chain -(C 1 -C 8 ) Alkyl groups include, but are not limited to, -isopropyl, -sec-butyl, -isobutyl, -tert-butyl, -isopentyl, and -2-methylbutyl. In some aspects, the alkyl group can be unsubstituted. Optionally, the alkyl group can be substituted, for example with one or more groups.

[0019] Unless otherwise indicated, the term “alkylene” by itself or as part of another term generally refers to an alkylene group having the indicated number of carbon atoms, preferably 1 to 10 carbon atoms (-(C 1 -C 10 ) alkylene-) or preferably 1 to 8 carbon atoms (-(C 1 -C 8 ) alkylene-) and refers to a substituted or unsubstituted branched or straight-chain saturated hydrocarbon radical having two monovalent radical centers derived by the removal of two hydrogen atoms from the same or two different carbon atoms of a parent alkane. If the number of carbon atoms is not indicated, the alkylene group can have from 1 to 8 carbon atoms. Typical alkylene radicals include methylene (-CH 2 -), 1,2-ethylene (-CH 2 CH 2 -), 1,3-n-propylene (-CH 2 CH 2 CH 2 -), and 1,4-n-butylene (-CH 2 CH 2 CH 2 CH 2In some embodiments, the alkylene group can be unsubstituted. Optionally, the alkylene group can be substituted, for example with one or more groups.

[0020] Unless otherwise indicated, the term "alkenyl" by itself or as part of another term generally refers to a substituted or unsubstituted straight or branched chain unsaturated hydrocarbon having a double bond and the indicated number of carbon atoms; e.g., "-(C 2 -C 8 )alkenyl" or "-(C 2 -C 10 "-(C)alkenyl" refers to an alkenyl group having 2 to 8 or 2 to 10 carbon atoms, respectively. If the number of carbon atoms is not specified, the alkenyl group may have 2 to 8 carbon atoms. Representative -(C 2 -C 8 ) Alkenyl groups include, but are not limited to, -ethenyl, -1-propenyl, -2-propenyl, -1-butenyl, -2-butenyl, -isobutenyl, -1-pentenyl, -2-pentenyl, -3-methyl-1-butenyl, -2-methyl-2-butenyl, and -2,3-dimethyl-2-butenyl. In some aspects, an alkenyl group can be unsubstituted. Optionally, an alkenyl group can be substituted, for example, with one or more groups.

[0021] Unless otherwise indicated, the term “alkenylene,” by itself or as part of another term, generally refers to an alkenylene having the indicated number of carbon atoms, preferably 2 to 10 carbon atoms (—(C 2 -C 10 ) alkenylene-) or preferably 2 to 8 carbon atoms (-(C 2 -C 8) alkenylene-) refers to a substituted or unsubstituted unsaturated branched or straight chain hydrocarbon radical having a double bond and having two monovalent radical centers derived by removing two hydrogen atoms from the same or two different carbon atoms of a parent alkene. If the number of carbon atoms is not indicated, the alkenylene group can have 1 to 8 carbon atoms. Typical alkenylene radicals include, but are not limited to, -ethenylene-, -1-propenylene-, 2-propenylene-, -1-butenylene-, -2-butenylene-, -isobutenylene-, -1-pentenylene-, -2-pentenylene-, -3-methyl-1-butenylene-, -2-methyl-2-butenylene-, and -2,3-dimethyl-2-butenylene-. In some aspects, the alkenylene group can be unsubstituted. Optionally, the alkenylene group can be substituted, for example with one or more groups.

[0022] Unless otherwise indicated, the term "alkynyl" by itself or as part of another term generally refers to a substituted or unsubstituted straight or branched chain unsaturated hydrocarbon having a triple bond and the indicated number of carbon atoms; e.g., "-(C 2 -C 8 )alkynyl" or "-(C 2 -C 10 "-(C)alkynyl" refers to an alkynyl group having 2 to 8 or 2 to 10 carbon atoms, respectively. If the number of carbon atoms is not specified, the alkynyl group may have 2 to 8 carbon atoms. Representative -(C 2 -C 8 Alkynyl groups include, but are not limited to, -acetylenyl, -1-propynyl, -2-propynyl, -1-butynyl, -2-butynyl, -1-pentynyl, -2-pentynyl, and -3-methyl-1-butynyl. In some aspects, an alkynyl group can be unsubstituted. Optionally, an alkynyl group can be substituted, for example, with one or more groups.

[0023] Unless otherwise indicated, the term “alkynylene,” by itself or as part of another term, generally refers to an alkynylene having the indicated number of carbon atoms, preferably 2 to 10 carbon atoms (—(C 2 -C 10 )alkynylene-) or preferably 2 to 8 carbon atoms (-(C 2 -C 8 )alkynylene-) refers to a substituted or unsubstituted branched or straight chain unsaturated hydrocarbon radical having a triple bond and having two monovalent radical centers derived by removing two hydrogen atoms from the same or two different carbon atoms of a parent alkyne. If the number of carbon atoms is not indicated, the alkynylene group can have 2 to 8 carbon atoms. Typical alkynylene radicals include, but are not limited to, -ethynylene-, -1-propynylene-, -2-propynylene-, -1-butynylene-, -2-butynylene-, -1-pentynylene-, -2-pentynylene-, and -3-methyl-1-butynylene-. In some aspects, the alkynylene group can be unsubstituted. Optionally, the alkynylene group can be substituted, for example with one or more groups.

[0024] Unless otherwise indicated, the term "aryl" by itself or as part of another term generally refers to a substituted or unsubstituted monovalent carbocyclic aromatic hydrocarbon radical of 6 to 20 carbon atoms (preferably 6 to 14 carbon atoms, more preferably 6 to 10 carbon atoms, and in a highly preferred embodiment 6 carbon atoms) derived by removing one hydrogen atom from a single carbon atom of a parent aromatic ring system. Some aryl groups are represented in the exemplary structures as "Ar". Typical aryl groups include, but are not limited to, radicals derived from benzene, substituted benzene, naphthalene, anthracene, and biphenyl. An exemplary aryl group is a phenyl group. In some aspects, the aryl group can be unsubstituted. Optionally, the aryl group can be substituted, for example with one or more groups.

[0025] Unless otherwise indicated, the term “arylene,” by itself or as part of another term, generally refers to an aryl group in which one of the hydrogen atoms has been replaced with a bond (i.e., it is divalent), with phenyl as an exemplary group, having the following structure: As shown in TIFF2024540691000037.tif22128, it is an aryl group as defined above, which may be in para, meta, or ortho orientation. In selected embodiments, for example, when the parallel connector unit comprises an arylene, the arylene is an aryl group as defined above, in which two or more of the hydrogen atoms of the aryl group are replaced with bonds (i.e., the arylene can be trivalent). In some aspects, the arylene group can be unsubstituted. Optionally, the alkynylene group can be substituted, for example, with one or more groups.

[0026] Unless otherwise indicated, the term "heterocycle" or "heterocyclic ring", by itself or as part of another term, generally refers to a heterocycle having the indicated number of carbon atoms (e.g., "(C 3 -C 8 ) Heterocycle" or "(C 3 -C 10 "Heterocycle" refers to a monovalent substituted or unsubstituted aromatic or non-aromatic monocyclic or bicyclic ring system having a heterocycle having 3-8 or 3-10 carbon atoms, respectively, and 1-4 heteroatom ring members independently selected from N, O, P, or S, and derived by removing one hydrogen atom from a ring atom of the parent ring system. One or more N, C, or S atoms in a heterocycle may be oxidized. The ring containing the heteroatom may be aromatic or non-aromatic. Unless otherwise specified, a heterocycle is attached to its pendant group at any heteroatom or carbon atom that results in a stable structure. (C 3 -C 8Representative examples of heterocycles include, but are not limited to, pyrrolidinyl, azetidinyl, piperidinyl, morpholinyl, tetrahydrofuranyl, tetrahydropyranyl, benzofuranyl, benzothiophene, indolyl, benzopyrazolyl, pyrrolyl, thiophenyl (thiophene), furanyl, thiazolyl, imidazolyl, pyrazolyl, pyrimidinyl, pyridinyl, pyrazinyl, pyridazinyl, isothiazolyl, and isoxazolyl. In some aspects, the heterocycle group can be unsubstituted. Optionally, the heterocycle group can be substituted, for example, with one or more groups.

[0027] Unless otherwise indicated, the term “heterocyclo” or “heterocyclic ring”, by itself or as part of another term, generally refers to a heterocyclic group, as defined above and having the indicated number of carbon atoms, in which one of the heterocyclic group's hydrogen atoms has been replaced with a bond (i.e., it is divalent). 3 -C 8 ) Heterocycle or (C 3 -C 10 ) heterocycle). In selected embodiments, for example, when the parallel connector unit includes a heterocycle, the heterocycle is a heterocyclic group as defined above, where two or more of the hydrogen atoms of the heterocycle group are replaced with bonds (i.e., the heterocycle can be trivalent). In some aspects, the heterocycle or heterocyclic ring can be unsubstituted. Optionally, the heterocycle or heterocyclic ring can be substituted, for example, with one or more groups.

[0028] Unless otherwise indicated, the term "carbocycle" or "carbocyclic ring", by itself or as part of another term, generally refers to a ring system having the indicated number of carbon atoms derived by the removal of one hydrogen atom from a ring atom of a parent ring system (e.g., "(C 3 -C 8 )Carbocyclic ring" or "(C 3 -C 10"Carbocycle" refers to a monovalent substituted or unsubstituted aromatic or non-aromatic monocyclic or bicyclic carbocyclic ring system having 3 to 8 or 3 to 10 carbon atoms, respectively. As illustrative but non-limiting examples, the carbocycle can be a 3-, 4-, 5-, 6-, 7- or 8-membered carbocycle. Representative (C 3 -C 8 ) Carbocyclic rings include, but are not limited to, phenyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclopentadienyl, cyclohexyl, cyclohexenyl, 1,3-cyclohexadienyl, 1,4-cyclohexadienyl, cycloheptyl, 1,3-cycloheptadienyl, 1,3,5-cycloheptatrienyl, cyclooctyl, and cyclooctadienyl. In some aspects, the carbocyclic ring can be unsubstituted. Optionally, the carbocyclic ring can be substituted, for example, with one or more groups.

[0029] Unless otherwise indicated, the term “carbocyclo” or “carbocyclic ring”, by itself or as part of another term, generally refers to a ring system having the indicated number of carbon atoms (e.g., “(C 3 -C 8 )Carbocyclo" or "(C 3 -C 10 )Carbocyclo" refers to a carbocyclic group as defined above having 3 to 8 or 3 to 10 carbon atoms, respectively. In selected embodiments, for example, when a parallel connector unit includes a carbocyclo or carbocyclic ring, the carbocyclo or carbocyclic ring is a carbocyclic group as defined above in which two or more of the carbocyclic group's hydrogen atoms are replaced with bonds (i.e., the carbocyclo or carbocyclic ring can be trivalent). In some aspects, the carbocyclo or carbocyclic ring can be unsubstituted. Optionally, the heterocyclo or heterocyclic ring can be substituted, for example, with one or more groups.

[0030] Unless otherwise indicated, the term “heteroalkyl”, by itself or in combination with another term, means an alkyl group having a designated number of carbon atoms, unless otherwise stated, such as (C 1 -C 8 )heteroalkyl or (C 1 -C 10 )heteroalkyl) and 1 to 10, preferably 1 to 3 heteroatoms selected from the group consisting of O, N, Si and S, and a stable straight or branched chain hydrocarbon, which is fully saturated or contains 1 to 3 degrees of unsaturation, or a combination thereof, where the nitrogen and sulfur atoms may be oxidized and the nitrogen heteroatom may be quaternized. The heteroatoms O, N and S may be located at any interior position of the heteroalkyl group or at the position at which the alkyl group is attached to the remainder of the molecule. The heteroatom Si may be located at any position of the heteroalkyl group, including the position at which the alkyl group is attached to the remainder of the molecule. Examples include -CH 2 -CH 2 -O-CH 3 , -CH 2 -CH 2 -NH-CH 3 , -CH 2 -CH 2 -N(CH 3 )-CH 3 , -CH 2 -S-CH 2 -CH 3 , -CH 2 -CH 2 -S(O)-CH 3 , -NH-CH 2 -CH 2 -NH-C(O)-CH 2 -CH 3 , -CH 2 -CH 2 -S(O) 2 -CH 3 , -CH=CH-O-CH 3 , -Si(CH 3 ) 3 , -CH 2 -CH=NO-CH 3 , and -CH=CH-N(CH 3)-CH 3 For example, -CH 2 -NH-OCH 3 and -CH 2 -O-Si(CH 3 ) 3 In a preferred embodiment, up to two heteroatoms may be consecutive, such as (C 1 -C 4 ) Heteroalkyl or heteroalkylene has 1 to 4 carbon atoms and 1 or 2 heteroatoms, (C 1 -C 3 ) Heteroalkyl or heteroalkylene has 1-3 carbon atoms and 1 or 2 heteroatoms. In some aspects, heteroalkyl or heteroalkylene is saturated. In some aspects, heteroalkyl or heteroalkylene can be unsubstituted. Optionally, heteroalkyl or heteroalkylene can be substituted, for example with one or more groups.

[0031] Unless otherwise stated, the term "heteroalkylene," by itself or as part of another substituent, includes the radical -CH 2 -CH 2 -S-CH 2 -CH 2 - and -CH 2 -S-CH 2 -CH 2 -NH-CH 2 - the indicated number of carbon atoms (e.g., (C 1 -C 8 )heteroalkylene or (C 1 -C 10)heteroalkylene) means a divalent group derived from a heteroalkyl (as described above). For heteroalkylene groups, heteroatoms can also occupy one or both of the chain termini. Furthermore, for alkylene and heteroalkylene linking groups, no orientation of the linking group is implied. In selected embodiments, for example, when the parallel connector unit comprises a heteroalkylene, the heteroalkylene is a heteroalkyl group as defined above, where two or more of the hydrogen atoms of the heteroalkyl group are replaced with bonds (i.e., the heteroalkylene can be trivalent). In some aspects, the heteroalkyl or heteroalkylene can be saturated. In some aspects, the heteroalkylene is unsubstituted. Optionally, the heteroalkylene can be substituted, for example, with one or more groups.

[0032] The term "halogen", unless otherwise defined, generally refers to elements of main group 7; preferably fluorine, chlorine, bromine and iodine; more preferably fluorine, chlorine and bromine; even more preferably fluorine and chlorine.

[0033] The terms "substituted," "optionally substituted," "optionally substituted," and the like, unless otherwise indicated, generally mean that one or more hydrogen atoms may each be independently replaced by a substituent. Exemplary substituents include -X, -R, -O, -C, -D, -E, -F ... - , -OR, -SR, -S - , -NR 2 , -NR 3 , =NR, -CX 3 , -CN, -OCN, -SCN, -N=C=O, -NCS, -NO, -NO 2 , =N 2 , -N 3 , -NRC(=O)R, -C(=O)R, -C(=O)NR 2 , -SO 3 - , -SO 3 H, -S(=O) 2 R, -OS(=O) 2 OR, -S(=O) 2NR, -S(=O)R, -OP(=O)(OR) 2 , -P(=O)(OR) 2 , -PO 4 3- , -PO 3 H 2 , -C(=O)R, -C(=O)X, -C(=S)R, -CO 2 R, -CO 2 H, -C(=S)OR, -C(=O)SR, -C(=S)SR, -C(=O)NR 2 , -C(=S)NR 2 , or -C(=NR)NR 2 where each X is independently a halogen: -F, -CI, -Br, or -I; and each R is independently -H, -(C 1 -C 20 ) alkyl (e.g., -(C 1 -C 10 ) alkyl or -(C 1 -C 8 ) alkyl), -(C 6 -C 20 ) aryl (e.g., -(C 6 -C 10 ) aryl or, preferably, -C 6 -aryl), -(C 3 -C 14 ) heterocycles (e.g., -(C 3 -C 10 ) heterocycle or -(C 3 -C 8 ) heterocycle), a protecting group, or a prodrug moiety. Typical substituents also include (=O).

[0034] The term "aliphatic or aromatic residue" as used herein generally refers to an aliphatic substituent, such as, but not limited to, an alkyl residue, which may, however, be substituted by further aliphatic and / or aromatic substituents. As a non-limiting example, an aliphatic residue may have a direct link (R 5In the case of, for example, the link to the nitrogen atom of Y) can be a nucleic acid, an enzyme, a coenzyme, a nucleotide, an oligonucleotide, a monosaccharide, a polysaccharide, a polymer, a fluorophore, an optionally substituted benzene, etc., so long as it is aliphatic. The aromatic residue is a substituent where the direct link to the core structure is an aromatic system, e.g., an optionally substituted phenyl or triazolyl or pyridyl, or part of a nucleotide; non-limiting examples include where the direct link of the nucleotide to the core structure is, for example, via a phenyl residue. The term "aromatic residue" as used herein also includes heteroaromatic residues.

[0035] The term "peptide", unless otherwise indicated, generally refers to an organic compound containing two or more amino acids covalently linked by peptide bonds (amide bonds). Peptides may be referred to in terms of the number of constituent amino acids, i.e., a dipeptide contains two amino acid residues, a tripeptide contains three, etc. Peptides containing ten or fewer amino acids are called oligopeptides, while those having ten or more amino acid residues, e.g., up to about 30 amino acid residues, are polypeptides.

[0036] The term "amino acid" as used herein generally refers to a -CH(NH 3 )-COOH group. In one embodiment, the term "amino acid" refers to naturally occurring amino acids. As illustrative examples, naturally occurring amino acids include arginine, lysine, aspartic acid, glutamic acid, glutamine, asparagine, histidine, serine, threonine, tyrosine, cysteine, methionine, tryptophan, alanine, isoleucine, leucine, phenylalanine, valine, proline and glycine. However, in a broader sense, this term also includes non-naturally occurring amino acids.

[0037] Amino acids and peptides according to the present disclosure can also be modified at functional groups. Non-limiting examples are sugars, such as N-acetylgalactosamine (GalNAc), or protecting groups, such as fluorenylmethoxycarbonyl (Fmoc) modifications or esters.

[0038] The term "antibody", as used herein, is preferably intended to refer to an immunoglobulin molecule composed of four polypeptide chains, two heavy (H) chains and two light (L) chains, typically interconnected by disulfide bonds. Each heavy chain is composed of a heavy chain variable region (abbreviated herein as VH) and a heavy chain constant region. The heavy chain constant region can, for example, contain three domains: CH1, CH2 and CH3. Each light chain is composed of a light chain variable region (abbreviated herein as VL) and a light chain constant region. The light chain constant region is composed of one domain (CL). The VH and VL regions can be further subdivided into regions of hypervariability, called complementarity determining regions (CDRs), interspersed with more conserved regions, called framework regions (FRs). Each VH and VL is typically composed of three CDRs and up to four FRs, arranged from amino-terminus to carboxy-terminus in the following order, for example: FR1, CDR1, FR2, CDR2, FR3, CDR3, FR4.

[0039] As used herein, the term "complementarity determining region" (CDR; e.g., CDR1, CDR2, and CDR3) refers to the amino acid residues of an antibody variable domain whose presence is necessary for antigen binding. Each variable domain typically has three CDR regions identified as CDR1, CDR2, and CDR3. Each complementarity determining region may comprise amino acid residues from a "complementarity determining region" as defined by Kabat (e.g., about residues 24-34 (L1), 50-56 (L2), and 89-97 (L3) in the light chain variable domain and 31-35 (H1), 50-65 (H2), and 95-102 (H3) in the heavy chain variable domain); and / or residues from a "hypervariable loop" (e.g., about residues 26-32 (L1), 50-52 (L2), and 91-96 (L3) in the light chain variable domain and 26-32 (H1), 53-55 (H2), and 96-101 (H3) in the heavy chain variable domain). In some cases, a complementarity determining region may comprise amino acids from both a CDR region and a hypervariable loop as defined according to Kabat.

[0040] Depending on the amino acid sequence of the constant domain of the heavy chain, intact antibodies can be assigned to different "classes". There are five major classes of intact antibodies: IgA, IgD, IgE, IgG, IgM, and some of these can be further divided into "subclasses" (isotypes), e.g., IgG1, IgG2, IgG3, IgG4, IgA1, and IgA2. The preferred class of immunoglobulin for use in the present invention is IgG.

[0041] The heavy chain constant domains corresponding to different classes of antibodies are called [alpha], [delta], [epsilon], [gamma] and [mu], respectively. The subunit structures and three-dimensional configurations of different classes of immunoglobulins are well known. As used herein, antibody refers to conventionally known antibodies and functional fragments thereof.

[0042] "Functional fragment" or "antigen-binding antibody fragment" of an antibody / immunoglobulin, or "antigen-binding fragment of an antibody", or "antibody fragment", or "fragment of an antibody" generally relates to a fragment of an antibody / immunoglobulin (e.g., the variable region of an IgG) that retains the antigen-binding region. The "antigen-binding region" of an antibody is typically found in one or more hypervariable regions of the antibody, e.g., the CDR1, -2, and / or -3 regions; however, variable "framework" regions may also play an important role in antigen binding, such as by providing a scaffold for the CDRs. Preferably, the "antigen-binding region" comprises at least amino acid residues 4-103 of the variable light (VL) chain and 5-109 of the variable heavy (VH) chain, more preferably amino acid residues 3-107 of the VL and 4-111 of the VH, with the complete VL and VH chains (amino acid positions 1-109 of the VL and 1-113 of the VH; numbering according to WO 97 / 08320).

[0043] "Functional fragments," "antigen-binding antibody fragments," "antigen-binding fragments of antibodies," or "antibody fragments" or "fragments of antibodies" of the present disclosure may include, but are not limited to, those that contain at least one disulfide bond that can react with a reducing agent as described herein. Examples of suitable fragments include Fab, Fab', Fab'-SH, F(ab') 2 F(ab') , and Fv fragments; diabodies; single domain antibodies (DAbs), linear antibodies; single chain antibody molecules (scFv); and multispecific antibodies such as bispecific and trispecific antibodies formed from antibody fragments. An antibody other than a "multispecific" or "multifunctional" antibody is understood to have each of its binding sites identical. F(ab') 2 Alternatively, the Fab can be engineered to minimize or completely eliminate intermolecular disulfide interactions that occur between the CH1 and CL domains.

[0044] The term "Fc region" herein is generally used to define the C-terminal region of an immunoglobulin heavy chain that includes at least a portion of the constant region. This term includes native sequence Fc regions and variant Fc regions. In one embodiment, a human IgG heavy chain Fc region extends from Cys226 or Pro230 to the carboxyl terminus of the heavy chain. However, the C-terminal lysine (Lys447) of the Fc region may or may not be present. Unless otherwise specified herein, the numbering of amino acid residues in the Fc region or constant region is according to the EU numbering system, also known as the EU index.

[0045] Variants of antibodies or antigen-binding antibody fragments contemplated herein are molecules that retain the binding activity of the antibody or antigen-binding antibody fragment.

[0046] "Binding proteins" or "proteinaceous binding molecules with antibody-like binding properties" as used herein are generally known to those skilled in the art. Illustrative, non-limiting examples include affibodies, adnectins, anticalins, DARPins, and avimers.

[0047] A "human" antibody or antigen-binding fragment thereof is generally defined as one that is not chimeric (e.g., not "humanized") and not derived (in whole or in part) from a non-human species. A human antibody or antigen-binding fragment thereof may be of human origin or may be a synthetic human antibody. A "synthetic human antibody" is defined herein as an antibody having a sequence derived in silico, in whole or in part, from a synthetic sequence based on the analysis of known human antibody sequences. In silico design of a human antibody sequence or fragment thereof can be achieved, for example, by analyzing a database of human antibody or antibody fragment sequences and devising a polypeptide sequence using data obtained therefrom. Another example of a human antibody or antigen-binding fragment thereof is one that is encoded by a nucleic acid isolated from a library of antibody sequences of human origin (e.g., such libraries are based on antibodies taken from human natural sources).

[0048] A "humanized antibody" or a humanized antigen-binding fragment thereof is generally defined herein as: (i) an antibody derived from a non-human source (e.g., a transgenic mouse with a xenogeneic immune system) based on human germline sequences; (ii) an antibody in which amino acids in the framework regions of a non-human antibody have been partially replaced with human amino acid sequences by genetic engineering; or (iii) a CDR-grafted antibody in which the CDRs of the variable domain are derived from a non-human source, but one or more frameworks of the variable domain are of human origin, and the constant domains (if present) are of human origin.

[0049] A "chimeric antibody" or antigen-binding fragment thereof is generally defined herein as one in which the variable domains are derived from a non-human source and some or all of the constant domains are derived from a human source.

[0050] The term "monoclonal antibody", as used herein, generally refers to an antibody obtained from a population of substantially homogeneous antibodies, i.e., the individual antibodies constituting the population are identical except for possible mutations that may be present in minor amounts, e.g., naturally occurring mutations. Thus, the term "monoclonal" indicates the property of an antibody that it is not a mixture of individual antibodies. In contrast to polyclonal antibody preparations, which typically contain different antibodies directed against different determinants (epitopes), each monoclonal antibody of a monoclonal antibody preparation is directed against a single determinant on an antigen. In addition to their specificity, monoclonal antibody preparations are advantageous in that they are typically not contaminated by other immunoglobulins. The term "monoclonal" should not be construed as requiring production of the antibody by any particular method. The term monoclonal antibody specifically includes chimeric, humanized, and human antibodies.

[0051] An "isolated" antibody is generally one that has been identified and separated from components of the cell in which it is expressed. The contaminating components of the cell are substances that would interfere with diagnostic or therapeutic uses of the antibody, and may include enzymes, hormones, and other proteinaceous or nonproteinaceous solutes.

[0052] As used herein, an antibody "specifically binds to," "is specific for," or "specifically recognizes" an antigen of interest, e.g., a tumor-associated polypeptide antigen target, and generally binds the antigen with sufficient affinity so that the antibody is useful as a therapeutic agent in targeting cells or tissues expressing the antigen, and does not significantly cross-react with other proteins or proteins other than orthologs and variants (e.g., mutant forms, splice variants, or proteolytically truncated forms) of the aforementioned antigen target. The terms "specifically recognize" or "specifically bind to" or "is specific for" a particular polypeptide or epitope on a particular polypeptide target as used herein refer to, for example, an antibody that "specifically binds to," "is specific for," or "is specific for," an antibody that "specifically recognizes" an antigen of interest, e.g., a tumor-associated polypeptide antigen target, and generally binds to the antigen with sufficient affinity so that the antibody is useful as a therapeutic agent in targeting cells or tissues expressing the antigen, and does not significantly cross-react with other proteins other than orthologs and variants (e.g., mutant forms, splice variants, or proteolytically truncated forms) of the aforementioned antigen target. -4 Less than M or about 10 -5 Less than M or about 10 -6 Less than M or about 10 -7 Less than M or about 10 -8 Less than M or about 10 -9 Less than M or about 10 -10 Less than M or about 10 -11 Less than M or about 10 -12 Monovalent K for antigens less than or equal to M DThe specific binding may be indicated by an antibody, or an antigen-binding fragment thereof, having a specific binding affinity of at least one of the following: A specific binding affinity may be indicated by an antibody, or an antigen-binding fragment thereof, having a specific binding affinity of at least one of the following: A specific binding affinity may be indicated by an antibody, or an antigen-binding fragment thereof, having a specific binding affinity of at least one of the following: A specific binding affinity may be indicated by an antibody, or an antigen-binding fragment thereof, having a specific binding affinity of at least one of the following: A specific binding affinity may be indicated by an antibody, or an antigen-binding fragment thereof, having a specific binding affinity of at least one of the following: A specific binding affinity may be indicated by an antibody, or an antigen-binding fragment thereof, having a specific binding affinity of at least one of the following: A specific binding affinity may be indicated by an antibody, or an antigen-binding fragment thereof, having a specific binding affinity of at least one of the following: A specific binding affinity may be indicated by an antibody, or an antigen-binding fragment thereof, having a specific binding affinity of at least one of the following: A specific binding affinity may be indicated by an antibody, or an antigen-binding fragment thereof, having a specific binding affinity of at least one of the following: This means that the positive / negative difference is 5-fold or more, 10-fold or more, 50-fold or more, and preferably 100-fold or more. Typically, the determination of binding specificity is not performed with a single reference antigen, but with a set of about 3-5 unrelated antigens, such as milk powder, BSA, transferrin, etc.

[0053] "Binding affinity" or "affinity" generally refers to the strength of the sum of non-covalent interactions between a single binding site of a molecule and its binding partner. Unless otherwise indicated, as used herein, "binding affinity" refers to the intrinsic binding affinity that reflects a 1:1 interaction between members of a binding pair (e.g., an antibody and an antigen). The dissociation constant "K D" is generally used to describe the affinity between a molecule (e.g., an antibody) and its binding partner (e.g., an antigen), i.e., how tightly a ligand binds to a particular protein. Ligand-protein affinity is influenced by non-covalent intermolecular interactions between the two molecules. Affinity can be measured by common methods known in the art, including those described herein. In one embodiment, "K" according to the present invention is D " or "K D The "value" is measured by using a surface plasmon resonance assay using a suitable instrument, including but not limited to a Biacore instrument such as the Biacore T100, Biacore T200, Biacore 2000, Biacore 4000, Biacore 3000 (GE Healthcare Biacore, Inc.), or a ProteOn XPR36 instrument (Bio-Rad Laboratories, Inc.).

[0054] The term "antibody drug conjugate" or abbreviated ADC is well known to those of skill in the art and, as used herein, generally refers to the linkage of an antibody or antigen-binding fragment thereof with a drug such as a chemotherapeutic agent, a toxin, an immunotherapeutic agent, an imaging probe, and the like.

[0055] The present disclosure also relates to "pharmaceutically acceptable salts". Any pharmaceutically acceptable salts can be used. In particular, the term "pharmaceutically acceptable salts" refers to salts of the conjugates or compounds of the present invention that are pharmaceutically acceptable and have the desired pharmacological activity of the parent compound. In particular, such salts have low toxicity and can be inorganic or organic acid addition salts and base addition salts. Specifically, such salts include (1) salts formed with inorganic acids such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, and the like; or salts formed with acetic acid, propionic acid, hexanoic acid, cyclopentanepropionic acid, glycolic acid, pyruvic acid, lactic acid, malonic acid, succinic acid, malic acid, maleic acid, fumaric acid, tartaric acid, citric acid, benzoic acid, 3-(4-hydroxybenzoyl)benzoic acid, cinnamic acid, mandelic acid, methanesulfonic acid, ethanesulfonic acid, 1,2-ethanedisulfonic acid, 2-hydroxyethanesulfonic acid, benzenesulfonic acid, 4-chlorobenzenesulfonic acid, 2-naphthalenesulfonic acid, 4-toluenesulfonic acid, camphorsulfonic acid, 4-methylbicyclo[2.2] ... or (2) salts formed when an acidic proton present in the parent compound is replaced by a metal ion, e.g., an alkali metal ion, an alkaline earth ion, or an aluminum ion; or when coordinated with an organic base, e.g., ethanolamine, diethanolamine, triethanolamine, N-methylglucamine, etc. Salts further include, purely by way of example, sodium, potassium, calcium, magnesium, ammonium, tetraalkylammonium, etc.; as well as, where the compound contains a basic functionality, salts of non-toxic organic or inorganic acids, e.g., hydrochloride, hydrobromide, tartrate, mesylate, acetate, maleate, oxalate, etc. To maintain electronic neutrality, counterions or anionic counterions can be used at the quaternary amines. Exemplary counterions include halide ions (e.g., F -, Cl - , Br - , I - ), NO 3 - , ClO 4 - , O.H. - , H 2 PO 4 - , HSO 4 - , sulfonate ions (e.g., methanesulfonic acid, trifluoromethanesulfonic acid, p-toluenesulfonic acid, benzenesulfonic acid, 10-camphorsulfonic acid, naphthalene-2-sulfonic acid, naphthalene-1-sulfonic acid-5-sulfonic acid, etc.), and carboxylate ions (e.g., acetic acid, ethanoic acid, propanoic acid, benzoic acid, glyceric acid, lactic acid, tartaric acid, glycolic acid, etc.).

[0056] As used herein, the term "solvate" may refer to an aggregate that includes one or more molecules of the conjugate or compound described herein and one or more molecules of a solvent. The solvent may be water, in which case the solvate may be a hydrate. Alternatively, the solvent may be an organic solvent. Thus, the conjugate or compound of the present disclosure may exist as a hydrate, including a monohydrate, a dihydrate, a hemihydrate, a sesquihydrate, a trihydrate, a tetrahydrate, and the like, as well as the corresponding solvated forms. The compounds of the present invention may be true solvates, while in other cases, the compounds of the present invention may simply retain adventitious water or may be a mixture of water and adventitious solvent.

[0057] Conjugates of formula (I) As indicated above, the present invention provides a conjugate having formula (I): TIFF2024540691000038.tif35128, or a pharma- ceutically acceptable salt or solvate thereof, During the ceremony: RBM is receptor binding molecule; TIFF2024540691000039.tif6128 is a double bond; or TIFF2024540691000040.tif6128 is a single bond; V is not present if TIFF2024540691000041.tif6128 is a double bond; or V is If TIFF2024540691000042.tif6128 is a single bond, it is H or (C 1 -C 8 ) alkyl; X is R if TIFF2024540691000043.tif6128 is a double bond 3 -C; or X is If TIFF2024540691000044.tif6128 is a single bond, TIFF2024540691000045.tif9128; Y is NR 5 , S, O, or CR 6 R 7 and; R 1 is a first polyalkylene glycol unit R comprising at least three alkylene glycol subunits; F and; R 3 is H; or an optionally substituted aliphatic or an optionally substituted aromatic residue; R 4 is H; or an optionally substituted aliphatic or an optionally substituted aromatic residue; R 5 is H; or an optionally substituted aliphatic or an optionally substituted aromatic residue; R 6 is H; or an optionally substituted aliphatic or an optionally substituted aromatic residue; R 7 is H; or an optionally substituted aliphatic or an optionally substituted aromatic residue; L is a linker; D is a drug moiety; m is an integer ranging from 1 to 10; and n is an integer ranging from 1 to 20.

[0058] The conjugate of formula (I) comprises a receptor-binding molecule linked to a drug moiety via a phosphorus (V) moiety (sometimes denoted as "P5") and a linker. 1 is a first polyalkylene glycol unit attached to an oxygen atom that is connected to the phosphorus atom of the phosphorus (V) moiety. Additionally, conjugates bearing a second polyalkylene glycol in the linker L, orthogonal to the orientation of the RBM and D, are also described.

[0059] The inventors have found that the conjugates of formula (I) have various advantageous properties, as shown below. The conjugates of formula (I) were prepared and tested with different linkers and drugs (the table in Example 2 provides an overview of the conjugates prepared and tested in Examples 3-7, for example, and conjugates with additional linkers and drugs are described in Examples 8-14). The conjugates of formula (I) were found to have good hydrophilicity and exhibit low aggregation in solution (Example 2 and Figures 13-30, Example 9 and Figures 45 and 46, Example 10 and Figure 47, Example 12 and Figure 50, and Example 14 and Figure 52). In addition, the conjugates of formula (I) exhibit good cytotoxicity against target-positive cancer cells (Example 4 and Figures 35-39, Example 8 and Figure 44, Example 13 and Figure 51, and Example 14 and Figure 53). The conjugates of formula (I) also show favorable bystander effects (Example 5 and Figure 40). Moreover, the conjugates of formula (I) show favorable in vivo efficacy, especially when compared directly with the efficacy of the marketed product ADCETRIS (Example 6 and Figure 41). Moreover, the conjugates of formula (I) also show good pharmacokinetic behavior in vivo, as illustrated by the very narrow concentration course of total antibody and intact ADC quantification over time, clearly indicating a very stable conjugate in vivo. The stability exceeds that of the marketed product ADCETRIS. Moreover, the ADC clearance from the blood circulation for the DAR8 VC-PAB-MMAE construct is not enhanced, being only similar to ADCETRIS with a DAR4 (Example 7 and Figure 42). In addition, the conjugates of formula (I) can be efficiently prepared with various ratios of drug moiety to receptor binding molecule (Examples 2 and 3 and Figure 31, Example 9 and Figure 45, Example 12 and Figure 49, and Example 14 and Figure 52). The conjugates of formula (I) can also be prepared with polyalkylene glycol units of various chain lengths (Examples 2 and 3 and Figure 32, and Example 8 and Figure 43).In this context, the inventors have also found that the efficiency, particularly the yield and drug-to-antibody ratio (DAR), of the conjugation reaction between the receptor-binding molecule and the linker-drug molecule resulting in the conjugates of formula (I) can be improved when longer PEG residues are used (as illustrative examples, PEG12 with 12 PEG units at the phosphorus atom, and even more so when PEG24 with 24 PEG units at the phosphorus atom is used) (Example 8 and Figure 43). In summary, the inventors have surprisingly found that the conjugates of formula (I) exhibit excellent properties that make them useful as pharmaceuticals, such as, for example, good efficiency of the conjugation reaction, good cytotoxicity against target-positive cancer cells, favorable bystander effect, excellent in vivo efficacy and in vivo pharmacokinetics. It is noted that conjugates comprising a phosphorus(V) moiety and a drug moiety are described, for example, in WO 2018 / 041985 A1, WO 2019 / 170710, and Kasper et al., Angew. Chem. Int. Ed. 2019, vol. 58, pp. 11631 to 11636, which are incorporated herein by reference.

[0060] Preferably R 3 is H or (C 1 -C 8 ) alkyl; more preferably R 3 is H. Preferably, R 4 If present, H or (C 1 -C 8 ) alkyl; more preferably R 4 When present, it is H. Preferably, R 5 If present, H or (C 1 -C 8 ) alkyl; more preferably R 5 When present, it is H. Preferably, R 6 If present, H or (C 1 -C 8 ) alkyl; more preferably R 6 When present, it is H. Preferably, R7 If present, H or (C 1 -C 8 ) alkyl; more preferably R 7 is H, if present.

[0061] Preferably, TIFF2024540691000046.tif6128 is a double bond; V is absent; X is R 3 -C; and R 3 is H or an optionally substituted aliphatic or an optionally substituted aromatic residue; preferably R 3 is H or (C 1 -C 8 ) alkyl; more preferably R 3 is H.

[0062] More preferably, TIFF2024540691000047.tif6128 represents a double bond; V is absent; X is R 3 -C and R 3 is H or (C 1 -C 8 ) alkyl. Preferably, R 3 is H or (C 1 -C 6 ) alkyl, more preferably H or (C 1 -C 4 ) alkyl, even more preferably H or (C 1 -C 2 ) alkyl. In a preferred embodiment, R 3 is H.

[0063] In some embodiments, TIFF2024540691000048.tif6128 can be a single bond; V can be H or (C 1 -C 8 ) alkyl, preferably V is H; X is TIFF2024540691000049.tif9128;R 3is H or an optionally substituted aliphatic or an optionally substituted aromatic residue; more preferably R 3 is H or (C 1 -C 8 ) alkyl, more preferably R 3 is H;R 4 is H or an optionally substituted aliphatic or an optionally substituted aromatic residue; preferably, R 4 is H or (C 1 -C 8 ) alkyl, preferably R 4 is H.

[0064] In some embodiments, TIFF2024540691000050.tif6128 can represent a bond; V can represent H or (C 1 -C 8 ) alkyl; X can be Represents TIFF2024540691000051.tif9128; and R 3 and R 4 are independently H or (C 1 -C 8 ) alkyl. Preferably, R 3 and R 4 are independently H or (C 1 -C 6 ) alkyl, more preferably H or (C 1 -C 4 ) alkyl, even more preferably H or (C 1 -C 2 ) alkyl. Preferably, R 3 and R 4 are the same; even more preferably, R 3 , R 4 and V are the same. More preferably, R 3 and R 4 are both H. Preferably, V is H or (C 1 -C 6 ) alkyl, more preferably H or (C 1 -C 4) alkyl, even more preferably H or (C 1 -C 2 ) alkyl. Even more preferably, V is H. In a preferred embodiment, R 3 , R 4 and V are each H.

[0065] The integer m ranges from 1 to 10. Thus, the integer m can be 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10. Preferably, the integer m ranges from 1 to 4. More preferably, the integer m is 1 or 2. Even more preferably, the integer m is 1.

[0066] The integer n is in the range of 1 to 20. Thus, the integer n can be 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19 or 20. Preferably, the integer n is in the range of 1 to 10. More preferably, the integer n is in the range of 2 to 10. Even more preferably, the integer n is in the range of 4 to 10. Even more preferably, the integer n is in the range of 6 to 10. Even more preferably, the integer n is 6, 7, 8, 9 or 10. Even more preferably, the integer n is in the range of 7 to 10. Even more preferably, the integer n is 7, 8 or 9. Even more preferably, the integer n is 7 or 8. Even more preferably, the integer n is in the range of 7 to 10. Even more preferably, the integer n is 8.

[0067] The integer n is in the range of 1 to 20. Preferably, the integer n is in the range of 1 to 10. More preferably, the integer n is in the range of 2 to 8. Even more preferably, the integer n is 2, 3, 4, 5 or 6. Even more preferably, the integer n is in the range of 3 to 6. Even more preferably, the integer n is 3, 4 or 5. Even more preferably, the integer n may be 4 or 5. Even more preferably, the integer n is 4.

[0068] Preferably, m is an integer in the range of 1 to 4, more preferably 1 or 2, even more preferably 1; and preferably n is an integer in the range of 1 to 20, more preferably 1 to 10, even more preferably 2 to 10, even more preferably 4 to 10, even more preferably 6 to 10, even more preferably n is 6, 7, 8, 9 or 10, even more preferably n is 7 to 10, even more preferably 7 to 10; even more preferably n is 7, 8 or 9, even more preferably n is 7 or 8, and still more preferably n is 8.

[0069] Preferably, m is an integer in the range of 1 to 4, more preferably m is 1 or 2, even more preferably m is 1; and preferably n is an integer in the range of 1 to 20, more preferably 1 to 10, even more preferably 2 to 8; even more preferably n is 2, 3, 4, 5 or 6; even more preferably n is in the range of 3 to 6; even more preferably n is 3, 4 or 5; even more preferably n is 4 or 5; and still more preferably n is 4.

[0070] Preferably, m is 1; and preferably, n is an integer ranging from 1 to 20, more preferably from 1 to 10, even more preferably from 2 to 10, even more preferably from 4 to 10, even more preferably from 6 to 10, even more preferably, n is 6, 7, 8, 9 or 10, even more preferably, n is in the range of 7 to 10, even more preferably, n is 7, 8 or 9, even more preferably, n is 7 or 8, and even more preferably, n is 8. Thus, preferably, m is 1 and n is an integer ranging from 1 to 20. More preferably, m is 1 and n is an integer ranging from 1 to 10. Even more preferably, m is 1 and n is an integer ranging from 2 to 10. Even more preferably, m is 1 and n is an integer ranging from 4 to 10. Even more preferably, m is 1 and n is an integer ranging from 6 to 10. Even more preferably, m is 1 and n is an integer ranging from 6 to 10. Even more preferably, m is 1 and n is 6, 7, 8, 9 or 10. Even more preferably, m is 1 and n is an integer in the range of 7 to 10. Even more preferably, m is 1 and n is 7, 8 or 9. Even more preferably, m is 1 and n is 7 or 8. Even more preferably, m is 1 and n is 8.

[0071] Preferably, m is 1; and in some embodiments, n is an integer ranging from 1 to 20, more preferably from 1 to 10, even more preferably from 2 to 8; even more preferably, n is 2, 3, 4, 5 or 6, even more preferably, n is in the range of 3 to 6; even more preferably, n is 3, 4 or 5; even more preferably, n is 4 or 5; and even more preferably, n is 4. Thus, preferably, m is 1 and n is an integer ranging from 1 to 20. More preferably, m is 1 and n is an integer ranging from 1 to 10. Even more preferably, m is 1 and n is an integer ranging from 2 to 8. Even more preferably, m is 1 and n is 2, 3, 4, 5 or 6. Even more preferably, m is 1 and n is in the range of 3 to 6. Even more preferably, m is 1 and n is 3, 4 or 5. Even more preferably, m is 1 and n is 4 or 5. Even more preferably, m is 1 and n is 4.

[0072] Preferably, the number of drug moieties D per receptor-binding molecule may be 1 to 20. More preferably, the number of drug moieties D per receptor-binding molecule is 1 to 14. Even more preferably, the number of drug moieties D per receptor-binding molecule is 2 to 14. Even more preferably, the number of drug moieties D per receptor-binding molecule is 4 to 14. Even more preferably, the number of drug moieties D per receptor-binding molecule is 5 to 12. Even more preferably, the number of drug moieties D per receptor-binding molecule is 6 to 12. Even more preferably, the number of drug moieties D per receptor-binding molecule is 7 to 10. Even more preferably, the number of drug moieties D per receptor-binding molecule is 8.

[0073] Preferably, the number of drug moieties D per receptor-binding molecule may be 1 to 20. More preferably, the number of drug moieties D per receptor-binding molecule is 1 to 14. Even more preferably, the number of drug moieties D per receptor-binding molecule is 1 to 12. Even more preferably, the number of drug moieties D per receptor-binding molecule is 2 to 10. Even more preferably, the number of drug moieties D per receptor-binding molecule is 2 to 8. Even more preferably, the number of drug moieties D per receptor-binding molecule is 2 to 6. Even more preferably, the number of drug moieties D per receptor-binding molecule is 3 to 5. Even more preferably, the number of drug moiety D populations per receptor-binding molecule is 4.

[0074] Receptor-binding molecule (RBM) RBM is a receptor binding molecule. The term "receptor binding molecule" generally refers to any molecule that can bind to a receptor. As an illustrative but non-limiting example, the receptor that the receptor binding molecule can bind to may be expressed on the cell surface. As an illustrative but non-limiting example, the cell expressing the receptor may be a cancer cell. Those skilled in the art know how to select a suitable receptor binding molecule.

[0075] The receptor may be a tumor-associated surface antigen. Thus, the receptor-binding molecule can specifically bind to the tumor-associated surface antigen. The term "tumor-associated surface antigen" as used herein generally refers to an antigen that is or can be presented on a surface located on or within a tumor cell. These antigens may be presented on the cell surface by an extracellular portion, which is often combined with a transmembrane and cytoplasmic portion of the molecule. These antigens may, in some embodiments, be presented only by tumor cells and not by normal, i.e., non-tumor cells. Tumor antigens may be expressed only on tumor cells or may exhibit tumor-specific mutations compared to non-tumor cells. In such embodiments, each antigen may be referred to as a tumor-specific antigen. Some antigens may be presented by both tumor cells and non-tumor cells and may be referred to as tumor-associated antigens. These tumor-associated antigens may be overexpressed in tumor cells compared to non-tumor cells and are accessible to antibody binding on tumor cells due to the less compact structure of tumor tissue compared to non-tumor tissue. In some embodiments, the tumor-associated surface antigen is located on the vasculature of the tumor. Illustrative but non-limiting examples of tumor-associated surface antigens include CD19, CD30, Her2 or PMSA. Tumor-associated surface antigens are known to those skilled in the art. In particular, those that have been found to be useful for the development of ADCs are described in the review by, for example, Criscitello et al., "Antibody-drug conjugates in solid tumors: a look into novel targets", Journal of Hematology and Oncology, (2021) 14:20 (https: / / doi.org / 10.1186 / s13045-021-01035-z).

[0076] The receptor binding molecule may be selected from the group consisting of antibodies, antibody fragments, and proteinaceous binding molecules with antibody-like binding properties.

[0077] Preferably, the receptor binding molecule is an antibody. More preferably, the antibody is selected from the group consisting of monoclonal antibodies, chimeric antibodies, humanized antibodies, human antibodies, and single domain antibodies, such as camel or shark single domain antibodies. Even more preferably, the antibody is a monoclonal antibody. Preferably, the antibody is capable of specifically binding to a tumor-associated surface antigen. In some embodiments, the antibody can be brentuximab. In some embodiments, the antibody can be trastuzumab.

[0078] The receptor binding molecule may be an antibody fragment. Preferably, the antibody fragment is a bivalent antibody fragment. More preferably, the bivalent antibody fragment is a (Fab) 2 Preferably, the antibody fragment is selected from the group consisting of a Fab fragment, a Fv fragment, and a single chain Fv fragment (scFv). It is also possible for the monovalent antibody fragment to be a fragment of a single domain camelid or shark single domain antibody. Preferably, the antibody fragment is capable of specifically binding to a tumor-associated surface antigen.

[0079] The receptor binding molecule may be a proteinaceous binding molecule with antibody-like binding properties. Examples of proteinaceous binding molecules with antibody-like binding properties that can be used as receptor binding molecules include aptamers, muteins based on the lipocalin family of polypeptides, glubodies, proteins based on ankyrin scaffolds, proteins based on crystallin scaffolds, adnectins, avimers, EGF-like domains, kringle domains, fibronectin type I domains, fibronectin type II domains, fibronectin type III domains, PAN domains, G1a domains, SRCR domains, Kunitz / bovine pancreatic trypsin inhibitor. domain, tendamistat, Kazal-type serine protease inhibitor domain, trefoil (P-type) domain, von Willebrand factor type C domain, anaphylatoxin-like domain, CUB domain, thyroglobulin type I repeat, LDL receptor class A domain, sushi domain, link domain, thrombospondin type I domain, immunoglobulin domain or immunoglobulin-like domain (e.g., domain antibody or camel heavy chain antibody), C-type lectin domain, MAM domain, von Willebrand factor type A domain, somatomedin B domain, WAP-type 4-disulfide core domain, F5 / 8 C-type domain, hemopexin domain, SH2 domain, SH3 domain, laminin-type EGF-like domain, C2 domain, "kappabody" (Ill. et al. "Design and construction of a hybrid immunoglobulin domain with properties of both heavy and light chain variable regions" Protein Eng 10:949-57 (1997)), "minibody" (Martin et al. "The affinity-selection of a minibody polypeptide inhibitor of human interleukin-6" EMBO J 13:5303-9 (1994)), "Janusin" (Traunecker et al."Bispecific single chain molecules (Janusins) target cytotoxic lymphocytes on HIV infected cells" EMBO J 10:3655-3659 (1991) and Traunecker et al. "Janusin: new molecular design for bispecific reagents" Int J Cancer Suppl 7:51-52 (1992), nanobodies, adnectins, tetranectins, microbodies, affilins, affibodies or ankyrins, crystallins, knottins, ubiquitins, zinc finger proteins, autofluorescent proteins, ankyrins or ankyrin repeat proteins or leucine-rich repeat proteins, avimers (Silverman, Lu Q, Bakker A, To W, Duguay A, Alba BM, Smith R, Rivas A, Li P, Le H, Whitehorn E, Moore KW, Swimmer C, Perlroth V, Vogt M, Kolkman J, Stemmer WP 2005, Nat Biotech, Dec;23(12):1556-61, E-Publication in Nat Biotech. 2005 Nov 20 edition); and Silverman J, Lu Q, Bakker A, To W, Duguay A, Alba BM, Smith R, Rivas A, Li P, Le H, Whitehorn E, Moore KW, Swimmer C, Perlroth V, Vogt M, Kolkman J, Stemmer WP, Nat Biotech, Dec;23(12):1556-61, E-Publication in Nat. Biotechnology.and multivalent avimer proteins evolved by exon shuffling of human receptor domain families, as described in the 2005 Nov 20 edition. Preferably, the proteinaceous binding molecule with antibody-like binding properties is selected from the group consisting of muteins based on polypeptides of the lipocalin family, glubodies, proteins based on ankyrin scaffolds, proteins based on crystallin scaffolds, adnectins, avimers, DARPins, and affibodies. Preferably, the proteinaceous binding molecule with antibody-like binding properties is capable of specifically binding to tumor-associated surface antigens.

[0080] Base Y The group Y is NR 5 , S, O, and C.R. 6 R 7 R 5 is H; or an optionally substituted aliphatic or an optionally substituted aromatic residue; preferably R 5 is H or (C 1 -C 8 ) alkyl; more preferably R 5 is H. R 6 is H; or an optionally substituted aliphatic or an optionally substituted aromatic residue; preferably R 6 is H or (C 1 -C 8 ) alkyl; more preferably R 6 is H. R 7 is H; or an optionally substituted aliphatic or an optionally substituted aromatic residue; preferably R 7 is H or (C 1 -C 8 ) alkyl; more preferably R 7 is H.

[0081] Preferably, Y is selected from the group consisting of NH, S, O and CH 2 More preferably, Y is NH, S or O. In some embodiments, Y is selected from the group consisting of CH 2In some embodiments, Y is O. In some embodiments, Y is S.

[0082] In a highly preferred embodiment, Y is NH.

[0083] base R 1: First polyalkylene glycol unit R F R 1 is the first polyalkylene glycol unit R F The term "first polyalkylene glycol unit," as used herein, refers to a polyalkylene glycol unit bonded to an O atom that is connected to the phosphorus of the phosphorus (V) moiety. F comprises at least three alkylene glycol subunits. Preferably, the first polyalkylene glycol unit R F has the following structure: More preferably, the first polyalkylene glycol unit R F has the following structure: TIFF2024540691000053.tif15128. Thus, the first polyalkylene glycol unit R F may be a polytetramethylene glycol unit, a polypropylene glycol unit, or a polyethylene glycol unit. Even more preferably, the first polyalkylene glycol unit R F has the following structure: Contains three or more alkylene glycol subunits having TIFF2024540691000054.tif14128.

[0084] Preferably, the first polyalkylene glycol unit R F More preferably, the first polyalkylene glycol unit RF comprises 3 to 50 alkylene glycol subunits as described herein. Even more preferably, the first polyalkylene glycol unit R F comprises 3 to 45 alkylene glycol subunits as described herein. Even more preferably, the first polyalkylene glycol unit R F comprises 4 to 40 alkylene glycol subunits as described herein. Even more preferably, the first polyalkylene glycol unit R F comprises 6 to 35 alkylene glycol subunits as described herein. Even more preferably, the first polyalkylene glycol unit R F contains 8 to 30 alkylene glycol subunits as described herein.

[0085] Preferably, the first polyalkylene glycol unit R F More preferably, the first polyalkylene glycol unit R F comprises 3 to 12 alkylene glycol subunits as described herein. Even more preferably, the first polyalkylene glycol unit R F contains 3 to 11 alkylene glycol subunits as described herein.

[0086] First polyalkylene glycol unit R F The structure: The first polyalkylene glycol unit R may be a polyalkylene glycol unit containing 3 to 100, preferably 3 to 50, more preferably 3 to 45, even more preferably 4 to 40, even more preferably 6 to 35, and still more preferably 8 to 30 subunits having the TIFF2024540691000055.tif15128. F The structure: The first polyalkylene glycol unit R may be a polyalkylene glycol unit containing 3 to 100, preferably 3 to 50, more preferably 3 to 45, even more preferably 4 to 40, even more preferably 6 to 35, and still more preferably 8 to 30 subunits having the TIFF2024540691000056.tif15128. F The structure: In a highly preferred embodiment, the first polyalkylene glycol unit R F The structure: It may be a polyethylene glycol unit containing 3 to 100, preferably 3 to 50, more preferably 3 to 45, even more preferably 4 to 40, even more preferably 6 to 35, and still more preferably 8 to 30 subunits each having the structure TIFF2024540691000058.tif10128.

[0087] First polyalkylene glycol unit R F The structure: The first polyalkylene glycol unit R F The structure: The first polyalkylene glycol unit R F The structure: In a highly preferred embodiment, the first polyalkylene glycol unit R F The structure: It may be a polyethylene glycol unit containing 3 to 20, preferably 3 to 12, and more preferably 3 to 11 subunits each having the structure TIFF2024540691000062.tif10128.

[0088] Preferably, the first polyalkylene glycol unit R F teeth, TIFF2024540691000063.tif14128, Where: TIFF2024540691000064.tif10128 shows the position of O connected to phosphorus; K F is H or a first capping group; preferably K F is -H (hydrogen), -PO 3 H, -(C 1 -C 10 ) alkyl, -(C 1 -C 10 )Alkyl-SO 3 H, -(C 2 -C 10 )Alkyl-CO 2 H, -(C 2 -C 10 ) alkyl-OH, -(C 2 -C 10 )Alkyl-NH 2 , -(C 2 -C 10 )Alkyl-NH(C 1 -C 3 ) alkyl and -(C 2 -C 10 )Alkyl-N((C 1 -C 3 )Alkyl) 2 More preferably, K F is H; and o is an integer ranging from 3 to 100.

[0089] A "first capping group," as referred to herein, may be any moiety capable of functioning as an end group of a first polyalkylene glycol unit. Examples of first capping groups that may be used in the present disclosure include -PO 3 H, -(C 1 -C 10 ) alkyl, -(C 1 -C 10 )Alkyl-SO 3 H, -(C 2 -C 10 )Alkyl-CO 2 H, -(C 2 -C 10 ) alkyl-OH, -(C 2 -C 10 )Alkyl-NH 2 , -(C 2 -C 10 )Alkyl-NH(C 1 -C 3 ) alkyl and -(C 2 -C 10 )Alkyl-N((C 1 -C 3 )Alkyl) 2 In some embodiments, the first capping group is -(C 1 -C 10 ) alkyl, especially methyl.

[0090] Preferably, K F is H (hydrogen).

[0091] The integer o represents the repeating unit in the first polyalkylene glycol unit: TIFF2024540691000065.tif12128 number is displayed. The integer o may be in the range of 3 to 100. Preferably, o is in the range of 3 to 50. More preferably, o is in the range of 3 to 45. Even more preferably, o is in the range of 4 to 40. Even more preferably, o is in the range of 6 to 35. Even more preferably, o is in the range of 8 to 30. Even more preferably, o is in the range of 4 to 16. Even more preferably, o is in the range of 8 to 16. Even more preferably, o is 10, 11, 12, 13 or 14. Even more preferably, o is 11, 12 or 13. In a preferred embodiment, o is 12 or about 12. Even more preferably, o is in the range of 16 to 30. Even more preferably, o is in the range of 20 to 28. Even more preferably, o is 22, 23, 24, 25 or 26. Even more preferably, o is 23, 24 or 25. In a preferred embodiment, o is 24 or about 24. Preferably, the repeat unit is TIFF2024540691000066.tif11128. More preferably, the repeat unit is TIFF2024540691000067.tif12128. In the first polyalkylene glycol unit, the integer o can be in the range of 3 to 20. Preferably, o is in the range of 3 to 12. More preferably, o is in the range of 3 to 11. Preferably, the repeating unit is TIFF2024540691000068.tif11128. More preferably, the repeat unit is The file is TIFF2024540691000069.tif12128.

[0092] Preferably, the first polyalkylene glycol unit R F has the following structure: TIFF2024540691000070.tif10128, i.e., the subunit is designated as an ethylene glycol subunit. Thus, preferably, the first polyalkylene glycol unit is a first polyethylene glycol unit. The first polyethylene glycol unit comprises at least one ethylene glycol subunit.

[0093] Preferably, the first polyalkylene glycol unit R F The structure: The first polyethylene glycol unit may be a first polyethylene glycol unit containing 3 to 100, preferably 3 to 50, more preferably 3 to 45, even more preferably 4 to 40, even more preferably 6 to 35, and still more preferably 8 to 30 ethylene glycol subunits each having the formula TIFF2024540691000071.tif9128.

[0094] Preferably, the first polyalkylene glycol unit R F The structure: The first polyethylene glycol unit may be a first polyethylene glycol unit containing 3 to 20, preferably 3 to 12, and more preferably 3 to 11 ethylene glycol subunits each having the formula TIFF2024540691000072.tif9128.

[0095] Preferably, the first polyalkylene glycol unit R F The structure: a first polyethylene glycol unit having TIFF2024540691000073.tif15128; Where: TIFF2024540691000074.tif10128 shows the position of O connected to phosphorus; K F is H (hydrogen) or a first capping group as described herein; preferably K F -H, -PO 3 H, -(C 1 -C 10 ) alkyl, -(C 1-C 10 )Alkyl-SO 3 H, -(C 2 -C 10 )Alkyl-CO 2 H, -(C 2 -C 10 ) alkyl-OH, -(C 2 -C 10 )Alkyl-NH 2 , -(C 2 -C 10 )Alkyl-NH(C 1 -C 3 ) alkyl and -(C 2 -C 10 )Alkyl-N((C 1 -C 3 )Alkyl) 2 More preferably, K F is H; and o is an integer ranging from 3 to 100. The integer o represents the repeating unit in the first polyethylene glycol unit: TIFF2024540691000075.tif12128 number. The integer o may be in the range of 3 to 100. Preferably, o is in the range of 3 to 50. More preferably, o is in the range of 3 to 45. Even more preferably, o is in the range of 4 to 40. Even more preferably, o is in the range of 6 to 35. Even more preferably, o is in the range of 8 to 30. Even more preferably, o is in the range of 4 to 16. Even more preferably, o is in the range of 8 to 16. Even more preferably, o is 10, 11, 12, 13 or 14. Even more preferably, o is 11, 12 or 13. In a preferred embodiment, o is 12 or about 12. Even more preferably, o is in the range of 16 to 30. Even more preferably, o is in the range of 20 to 28. Even more preferably, o is 22, 23, 24, 25 or 26. Even more preferably, o is 23, 24 or 25. In a preferred embodiment, o is 24 or about 24.

[0096] In the first polyethylene glycol unit, the integer o can be in the range of 3 to 20. Preferably, o is in the range of 3 to 12. More preferably, o is in the range of 3 to 11.

[0097] Generally, the first polyalkylene glycol unit R F In the first polyalkylene glycol unit (preferably the first polyethylene glycol unit), polydisperse polyalkylene glycols (preferably polydisperse polyethylene glycols), monodisperse polyalkylene glycols (preferably monodisperse polyethylene glycols), and individual polyalkylene glycols (preferably individual polyethylene glycols) can be used. Polydisperse polyalkylene glycols (preferably polydisperse polyethylene glycols) are heterogeneous mixtures of sizes and molecular weights, whereas monodisperse polyalkylene glycols (preferably monodisperse polyethylene glycols) are typically purified from heterogeneous mixtures and thus provide a single chain length and molecular weight. The preferred first polyalkylene glycol unit is an individual polyalkylene glycol (preferably individual polyethylene glycol), i.e., a compound that is synthesized stepwise rather than by a polymerization process. An individual polyalkylene glycol (preferably individual polyethylene glycol) provides a single molecule with a defined and specified chain length.

[0098] The first polyalkylene glycol unit (preferably, the first polyethylene glycol unit) provided herein comprises one or more polyalkylene glycol chains (preferably, polyethylene glycol chains). The polyalkylene glycol chains (preferably, polyethylene glycol chains) can be linked together, for example, in a linear, branched, or star configuration. Optionally, at least one of the polyalkylene glycol chains (preferably, polyethylene glycol chains) can be derivatized at one end for covalent attachment to an oxygen atom attached to phosphorus.

[0099] The first polyalkylene glycol unit (preferably the first polyethylene glycol unit) is connected to the conjugate (or intermediate thereof) at an oxygen atom bonded to phosphorus. The other end(s) of the first polyalkylene glycol unit (preferably the first polyethylene glycol unit) is free and untethered and may take the form of hydrogen, methoxy, carboxylic acid, alcohol, or other suitable functional group, such as any first capping group, for example, as described herein. The methoxy, carboxylic acid, alcohol, or other suitable functional group acts as a cap for the terminal polyalkylene glycol subunit (preferably the polyethylene glycol subunit) of the first polyalkylene glycol unit (preferably the first polyethylene glycol unit). Untethered means that the first polyalkylene glycol unit (preferably the first polyethylene glycol unit) is not connected at its untethered site to a drug moiety (D), to a receptor binding molecule, or to a component of a linker (L) that links the drug moiety and / or the receptor binding molecule. For embodiments in which the first polyalkylene glycol unit (preferably the first polyethylene glycol unit) comprises more than one polyalkylene glycol chain (preferably a polyethylene glycol chain), the multiple polyalkylene glycol chains (preferably polyethylene glycol chains) may be the same or different chemical moieties (e.g., polyalkylene glycols, particularly polyethylene glycols, of different molecular weights or subunit numbers). The multiple first polyalkylene glycol chains (preferably the first polyethylene glycol chains) are attached at a single attachment site to the oxygen atom bonded to the phosphorus.One skilled in the art will appreciate that the first polyalkylene glycol unit (preferably a first polyethylene glycol unit), in addition to comprising repeating polyalkylene glycol subunits (preferably polyethylene glycol subunits), may further contain a non-polyalkylene glycol material (preferably a non-polyethylene glycol material) (e.g., to facilitate coupling of multiple polyalkylene glycol chains (preferably polyethylene glycol chains) to one another or to an oxygen atom attached to the phosphorus). The non-polyalkylene glycol material (preferably a non-polyethylene glycol material) comprises repeating alkylene glycol subunits (preferably -CH. 2 CH 2 The term "polyalkylene glycol" refers to an atom in the first polyalkylene glycol unit (preferably, the first polyethylene glycol unit) that is not part of a 1-O-subunit. In embodiments provided herein, the first polyalkylene glycol unit (preferably, the first polyethylene glycol unit) can include two monomeric polyalkylene glycol chains (preferably, polyethylene glycol chains) linked to each other via a non-polyalkylene glycol (non-polyethylene glycol) element. In other embodiments provided herein, the first polyalkylene glycol unit (preferably, the first polyethylene glycol unit) can include two linear polyalkylene glycol chains (preferably, polyethylene glycol chains) connected to a central core that is connected to an oxygen atom bonded to phosphorus (i.e., the polyalkylene glycol unit (preferably, the polyethylene glycol unit) is branched).

[0100] There are numerous methods of polyalkylene glycol (preferably polyethylene glycol) attachment available to one of skill in the art [see, for example, EP 0 401 384 (PEG coupling to G-CSF); U.S. Pat. No. 5,757,078 (PEGylation of EPO peptides); U.S. Pat. No. 5,672,662 (Polyethylene glycol monosubstituted with propionic or butanoic acid and their functional derivatives for biotechnological applications) and related polymers; U.S. Pat. No. 6,077,939 (PEGylation of the N-terminal α-carbon of peptides); and Veronese (2001) Biomaterials 22:405-417 (review on peptide and protein PEGylation)].

[0101] In preferred embodiments, the first polyalkylene glycol unit, more preferably the first polyethylene glycol unit, is directly connected to the oxygen atom attached to the phosphorus. In these embodiments, the first polyalkylene glycol unit, preferably the first polyethylene glycol unit, does not contain a functional group for connection to the oxygen atom attached to the phosphorus, i.e., the oxygen atom is not connected to a carbon atom of the first polyalkylene glycol unit, preferably to a CH of the first polyethylene glycol unit. 2 is directly coupled to

[0102] In one group of embodiments, the first polyalkylene glycol unit comprises at least 3 alkylene glycol subunits, even more preferably at least 4 alkylene glycol subunits, even more preferably at least 6 alkylene glycol subunits, and even more preferably at least 8 alkylene glycol subunits. In some such embodiments, the first polyalkylene glycol unit comprises no more than about 100 alkylene glycol subunits, preferably no more than about 50 alkylene glycol units, more preferably no more than about 45 alkylene glycol subunits, more preferably no more than about 40 alkylene glycol subunits, more preferably no more than about 35 subunits, and even more preferably no more than about 30 alkylene glycol subunits. In any one of these embodiments, the alkylene glycol subunits can be any alkylene glycol subunit as described herein. Preferably, in any one of these embodiments, each alkylene glycol subunit has the following structure: Preferably, when each alkylene glycol subunit is an ethylene glycol subunit, in any one of these embodiments the first polyalkylene glycol unit is a first polyethylene glycol unit.

[0103] In one group of embodiments, the first polyalkylene glycol unit comprises one or more linear polyalkylene glycol chains each having at least 3 alkylene glycol subunits, even more preferably at least 4 alkylene glycol subunits, even more preferably at least 6 alkylene glycol subunits, and even more preferably at least 8 alkylene glycol subunits. In preferred embodiments, the first polyalkylene glycol unit comprises a total of at least 3, even more preferably at least 4, even more preferably at least 6, or even more preferably at least 8 alkylene glycol subunits. In some such embodiments, the first polyalkylene glycol unit comprises a total of about 100 or less alkylene glycol subunits, preferably a total of about 50 or less alkylene glycol subunits, more preferably a total of about 45 or less subunits, even more preferably a total of about 40 or less subunits, even more preferably a total of about 35 or less subunits, and even more preferably a total of about 30 or less subunits. In any one of these embodiments, the alkylene glycol subunits can be any alkylene glycol subunit as described herein. Preferably, in any one of these embodiments, each alkylene glycol subunit has the following structure: TIFF2024540691000077.tif9128. Preferably, when each alkylene glycol subunit is an ethylene glycol subunit, in any one of these embodiments, the first polyalkylene glycol unit is a first polyethylene glycol unit that comprises one or more linear polyethylene glycol chains.

[0104] In another group of embodiments, the first polyalkylene glycol unit comprises a total of 3 to 100, preferably 3 to 50, more preferably 3 to 45, even more preferably 4 to 40, even more preferably 6 to 35, and still more preferably 8 to 30 alkylene glycol subunits. In any one of these embodiments, the alkylene glycol subunits can be any alkylene glycol subunit as described herein. Preferably, in any one of these embodiments, each alkylene glycol subunit has the following structure: Preferably, when each alkylene glycol subunit is an ethylene glycol subunit, in any one of these embodiments the first polyalkylene glycol unit is a first polyethylene glycol unit.

[0105] In another group of embodiments, the first polyalkylene glycol unit comprises one or more linear polyalkylene glycol chains having a total of 3 to 100, preferably 3 to 50, more preferably 3 to 45, even more preferably 4 to 40, even more preferably 6 to 35, and still more preferably 8 to 30 alkylene glycol subunits. In any one of these embodiments, the alkylene glycol subunits can be any alkylene glycol subunit as described herein. Preferably, in any one of these embodiments, each alkylene glycol subunit has the following structure: TIFF2024540691000079.tif10128. Preferably, when each alkylene glycol subunit is an ethylene glycol subunit, in any one of these embodiments the first polyalkylene glycol unit is a first polyethylene glycol unit comprising one or more linear polyethylene glycol chains.

[0106] In another group of embodiments, the first polyalkylene glycol unit is a linear single polyalkylene glycol chain having at least 3 subunits, even more preferably at least 6 subunits, and even more preferably at least 8 subunits. In any one of these embodiments, the alkylene glycol subunits can be any alkylene glycol subunit as described herein. Preferably, in any one of these embodiments, each alkylene glycol subunit has the following structure: TIFF2024540691000080.tif9128. Preferably, when each alkylene glycol subunit is an ethylene glycol subunit, in any one of these embodiments, the first polyalkylene glycol unit is a first polyethylene glycol unit that is a linear single polyethylene glycol chain. Optionally, in any one of these embodiments, the linear single polyalkylene glycol chain can be derivatized.

[0107] In another group of embodiments, the polyalkylene glycol unit is a linear single polyalkylene glycol chain having 3 to 100, preferably 3 to 50, more preferably 3 to 45, more preferably 4 to 40, more preferably 6 to 35, more preferably 8 to 30 alkylene glycol subunits. In any one of these embodiments, the alkylene glycol subunits can be any alkylene glycol subunit as described herein. Preferably, in any one of these embodiments, each alkylene glycol subunit has the following structure: TIFF2024540691000081.tif10128. Preferably, when each alkylene glycol subunit is an ethylene glycol subunit, in any one of these embodiments, the first polyalkylene glycol unit is a first polyethylene glycol unit that is a linear single polyethylene glycol chain. Optionally, in any one of these embodiments, the linear single polyalkylene glycol chain can be derivatized.

[0108] In any one of the embodiments provided herein, an exemplary linear polyethylene glycol unit that can be used as the first polyalkylene glycol unit, and in particular as the first polyethylene glycol unit, is as follows: TIFF2024540691000082.tif40128 where the wavy line indicates the site of connection to the oxygen atom bonded to phosphorus; R 20 is a PEG connecting unit; preferably, R 20 does not exist; R 21 is a PEG capping unit (herein R 21 "K F " is also displayed); R 22 is a PEG coupling unit (i.e., for coupling multiple PEG subunit chains together); n is independently selected from 3 to 100, preferably 3 to 50, more preferably 3 to 45, more preferably 4 to 40, even more preferably 6 to 35, and still more preferably 8 to 30; e is 2 to 5; Each n' is independently selected from 1 to 100, preferably 2 to 50, more preferably 3 to 45, more preferably 4 to 40, even more preferably 6 to 35, and still more preferably 8 to 30. In preferred embodiments, there are at least 3, more preferably at least 4, more preferably at least 6, and even more preferably at least 8 ethylene glycol subunits in the polyethylene glycol unit. In some embodiments, there are 100 or less, preferably 50 or less, more preferably 45 or less, more preferably 40 or less, more preferably 35 or less, and still more preferably 30 or less ethylene glycol subunits in the polyethylene glycol unit. R 20 If there is no (CH 2 CH 2 O) subunits are bonded directly to an oxygen atom that is bonded to phosphorus.

[0109] Preferably, the linear polyethylene glycol unit is TIFF2024540691000083.tif9128, where the wavy line indicates the site of attachment to the oxygen atom bonded to phosphorus; 20 , R 21 (As used herein, "K F "), and n are as defined herein; more preferably R 20 is absent. In a preferred embodiment, n is 12 or about 12. In a preferred embodiment, n is 24 or about 24. Preferably, R 21 is H.

[0110] Polyethylene glycol connecting unit R 20 When present, PEG-linking unit R is part of the first polyethylene glycol unit and acts to link the first polyethylene glycol unit to the oxygen atom attached to the phosphorus. In this regard, the oxygen atom attached to the phosphorus forms a bond with the first polyethylene glycol unit. In an exemplary embodiment, the PEG-linking unit R 20 If present, *-(C 1 -C 10 )Alkyl-# , *-Arylene- # , *-(C 1 -C 10 )Alkyl-O- # , *-(C 1 -C 10 )Alkyl-C(O)- # , *-(C 1 -C 10 )Alkyl-C(O)O- # , *-(C 1 -C 10 )Alkyl-NH- # , *-(C 1 -C 10 )Alkyl-S- # , *-(C 1 -C 10 )Alkyl-C(O)-NH- # , *-(C 1 -C 10 )Alkyl-NH-C(O)- # , and *-CH 2 -CH 2 SO 2 -(C 1 -C 10 )Alkyl- # where * denotes the point of attachment to the oxygen bonded to the phosphorus, and # denotes the point of attachment to the ethylene glycol unit.

[0111] PEG coupling unit R 22 is a part of the polyethylene glycol unit, if present, and is a repeating -CH 2 CH 2 A non-PEG material that acts to link two or more chains of O-subunits. In an exemplary embodiment, a PEG coupling unit R 22 If present, *-(C 1 -C 10 )Alkyl-C(O)-NH- # , *-(C 1 -C 10 )Alkyl-NH-C(O)- # , *-(C 2 -C 10 )Alkyl-NH- # , *-(C 2-C 10 )Alkyl-O- # , *-(C 1 -C 10 )Alkyl-S- # , or *-(C 2 -C 10 )Alkyl-NH- # where * denotes a point of attachment to an oxygen atom of the ethylene glycol subunit, and # denotes a point of attachment to a carbon atom of another ethylene glycol subunit.

[0112] As used herein, "K F ", also displayed as the group R 21 is H (hydrogen) in an exemplary embodiment, or may be a first capping group, as described herein; preferably, R 21 -H, -PO 3 H, -(C 1 -C 10 ) alkyl, -(C 1 -C 10 )Alkyl-SO 3 H, -(C 2 -C 10 )Alkyl-CO 2 H, -(C 2 -C 10 ) alkyl-OH, -(C 2 -C 10 )Alkyl-NH 2 , -(C 2 -C 10 )Alkyl-NH(C 1 -C 3 ) alkyl and -(C 2 -C 10 )Alkyl-N((C 1 -C 3 )Alkyl) 2 In some embodiments, R21 is independently selected from the group consisting of: 1 -C 10 ) alkyl, in particular methyl. More preferably, R 21 is H.

[0113] In any one of the embodiments provided herein, an illustrative linear first polyethylene glycol unit that can be used as the first polyalkylene glycol unit is as follows: and each n is 3 to 100, preferably 3 to 50, more preferably 3 to 45, even more preferably 4 to 40, even more preferably 6 to 35, and still more preferably 8 to 30. In some embodiments, n is about 12. In some embodiments, n is about 24.

[0114] In some embodiments, the first polyalkylene glycol unit is from about 300 daltons to about 5 kilodaltons; from about 300 daltons to about 4 kilodaltons; from about 300 daltons to about 3 kilodaltons; from about 300 daltons to about 2 kilodaltons; or from about 300 daltons to about 1 kilodalton. In some such aspects, the first polyalkylene glycol unit can have at least 6 alkylene glycol subunits or at least 8 alkylene glycol subunits. In some such aspects, the first polyalkylene glycol unit can have at least 6 alkylene glycol subunits or at least 8 alkylene glycol subunits, but not more than 100 alkylene glycol subunits, preferably not more than 50 alkylene glycol subunits. In some embodiments, the first polyalkylene glycol unit is a first polyethylene glycol unit that is about 300 daltons to about 5 kilodaltons; about 300 daltons to about 4 kilodaltons; about 300 daltons to about 3 kilodaltons; about 300 daltons to about 2 kilodaltons; or about 300 daltons to about 1 kilodalton. In some such aspects, the first polyethylene glycol unit can have at least 6 ethylene glycol subunits or at least 8 ethylene glycol subunits. In some such aspects, the first polyethylene glycol unit has at least 6 ethylene glycol subunits or at least 8 ethylene glycol subunits, but no more than 100 ethylene glycol subunits, preferably no more than 50 ethylene glycol subunits.

[0115] In some embodiments, R 1 is the first polyalkylene glycol unit R F When R is, there are no other alkylene glycol subunits present in the conjugate of formula (I) (i.e., no alkylene glycol subunits are present in any of the other components of the conjugate, such as, for example, in the linker L as provided herein). 1is the first polyalkylene glycol unit, then no more than 8, no more than 7, no more than 6, no more than 5, no more than 4, no more than 3, no more than 2, or no more than 1 other alkylene glycol subunits are present in the conjugate of Formula (I) (i.e., no more than 8, 7, 6, 5, 4, 3, 2, or 1 other alkylene glycol subunits are present in other components of the conjugate, such as, for example, in a linker L as provided herein).

[0116] Preferably, in another embodiment, R 1 is the first polyalkylene glycol unit R F the conjugate comprises a second polyalkylene glycol unit R, as described herein. S Preferably, R 1 is the first polyethylene glycol unit and the conjugate comprises a second polyalkylene glycol unit R S When the second polyalkylene glycol unit further comprises a second polyethylene glycol unit, as described herein.

[0117] It will be understood that in referring to alkylene glycol subunits, particularly ethylene glycol subunits, and depending on the context, the number of subunits may represent an average number, for example, when referring to a population of conjugates or intermediate compounds and using polydisperse polyalkylene glycols, particularly polydisperse polyethylene glycols.

[0118] "L": Linker The present disclosure provides a conjugate in which a receptor-binding molecule as described herein is linked to a drug moiety. According to the present disclosure, the receptor-binding molecule can be linked to the drug moiety via a covalent bond by a group Y and a linker L. As used herein, a "linker" L is any chemical moiety that can link a group Y, such as NH, to another moiety, such as a drug moiety. In this regard, reference is again made to formula (I) as described herein. Thus, the drug moiety D can be linked to Y via a linker L. In formula (I), RBM, TIFF2024540691000086.tif5128, V, X, Y, R 1 , L, D, m and n are as defined herein. The linker L serves to link Y with the drug moiety (D). The linker L is any chemical moiety capable of linking Y to the drug moiety D. In particular, the linker L connects Y to the drug moiety D by a covalent bond. A linker reagent is a bifunctional or polyfunctional moiety that can be used to link the drug moieties D and Y to form a conjugate of formula (I). The terms "linker reagent", "crosslinking reagent", "linker derived from a crosslinking reagent" and "linker" may be used interchangeably throughout this disclosure.

[0119] The linker is susceptible to cleavage, such as enzymatic cleavage, acid-induced cleavage, light-induced cleavage and disulfide bond cleavage (cleavable linker). Enzymatic cleavage includes, but is not limited to, protease-induced cleavage, peptidase-induced cleavage, esterase-induced cleavage, glycosidase-induced cleavage, phosphatase-induced cleavage, and sulfatase-induced cleavage, preferably under conditions in which the drug moiety and / or the receptor-binding molecule remain active. Alternatively, the linker can be substantially resistant to cleavage (e.g., stable linker or non-cleavable linker). In some aspects, the linker can be a procharged linker, a hydrophilic linker, a PEG-based linker, or a dicarboxylic acid-based linker. Thus, in some embodiments of any one of the antibody drug conjugates disclosed herein, the linker (L) is selected from the group consisting of a cleavable linker, a non-cleavable linker, a hydrophilic linker, a PEG-based linker, a procharged linker, a peptidic linker, and a dicarboxylic acid-based linker. Preferably, the linker L is a cleavable linker. In some embodiments, the linker L is a non-cleavable linker.

[0120] Preferably, the linker L is cleavable as described herein. In some embodiments, L is a linker susceptible to enzymatic cleavage. In some embodiments, L is an acid-labile linker, a photolabile linker, a peptidase-cleavable linker, a protease-cleavable linker, an esterase-cleavable linker, a glycosidase-cleavable linker, a phosphatase-cleavable linker, a sulfatase-cleavable linker, a disulfide bond-reducing linker, a hydrophilic linker, a precharged linker, a PEG-based linker, or a dicarboxylic acid-based linker. Preferably, the linker L is cleavable by a protease, a glucuronidase, a sulfatase, a phosphatase, an esterase, or by disulfide reduction. Preferably, the linker is a peptidase-cleavable linker. Other preferred linkers are cleavable by a protease.

[0121] A non-cleavable linker is any chemical moiety that can link a drug moiety to Y in a stable covalent manner and does not fall into the categories listed herein for cleavable linkers.Thus, a non-cleavable linker is substantially resistant to acid-induced cleavage, light-induced cleavage, peptidase-induced cleavage, protease-induced cleavage, glycosidase-induced cleavage, phosphatase-induced cleavage, esterase-induced cleavage and disulfide bond cleavage.Furthermore, non-cleavability refers to the ability of the chemical bond in or adjacent to the linker to withstand cleavage induced by acid, photolabile cleaving agent, peptidase, protease, glycosidase, phosphatase, esterase, or chemical or physiological compound that cleaves disulfide bonds under conditions that do not cause the drug moiety or receptor-binding molecule to lose its activity.

[0122] An acid-labile linker is a linker that is cleavable at an acidic pH. For example, certain intracellular compartments, such as endosomes and lysosomes, have an acidic pH (pH 4-5), providing suitable conditions for cleaving an acid-labile linker.

[0123] Some linkers can be cleaved by peptidases; i.e., peptidase-cleavable linkers. In this regard, certain peptides are easily cleaved intracellularly or extracellularly, see, for example, Trout et al., 79 Proc. Natl. Acad. Sci. USA, 626-629 (1982) and Umemoto et al. 43 Int. J. Cancer, 677-684 (1989). Peptides are composed of α-amino acids and peptide bonds, which are chemically amide bonds between the carboxylate of one amino acid and the amino group of another amino acid.

[0124] Some linkers can be cleaved by esterases; i.e., esterase-cleavable linkers. In this regard, certain esters can be cleaved by esterases present inside or outside the cell. Esters are formed by the condensation of a carboxylic acid with an alcohol. Simple esters are esters made with simple alcohols, such as aliphatic alcohols, and small cyclic and small aromatic alcohols.

[0125] Procharged linkers are derived from charged cross-linking reagents that retain their charge after incorporation into an antibody drug conjugate. Examples of procharged linkers can be found in US 2009 / 0274713.

[0126] Preferably, the linker L is cleavable as described herein. As illustrative examples, the linker may be cleavable by a protease, a glucuronidase, a sulfatase, a phosphatase, an esterase, or by disulfide reduction. Preferably, the linker L is cleavable by a protease. More preferably, the linker is cleavable by a cathepsin, in particular cathepsin B. The linker may include a dipeptide moiety, such as a valine-citrulline moiety or a valine-alanine moiety, which may be cleaved by a cathepsin, such as cathepsin B. Thus, in some embodiments, the linker includes a valine-citrulline moiety. In some embodiments, the linker includes a valine-alanine moiety. The linker may include a cleavage site. The term "cleavage site" may refer to a chemical moiety that is recognized by an enzyme and subsequently cleaved, for example by hydrolysis. As illustrative examples, the cleavage site is a sequence of amino acids that is recognized by a protease or peptidase and hydrolyzed by said protease or peptidase. In some embodiments, the cleavage site is a dipeptide. In some embodiments, the cleavage side is a valine-citrulline moiety. In some embodiments, the cleavage site is a valine-alanine moiety.

[0127] Second Spacer Unit In a preferred embodiment, the linker (L) comprises a second spacer unit -A- bonded to -Y-. The second spacer unit serves to link -Y- to another part of the linker (if present) or to the drug moiety (-D). As will be readily understood by those skilled in the art, this depends on whether another part of the linker is present. The second spacer unit (-A-) can be any chemical group or moiety that can link -Y- to another part of the linker, if present, or to the drug moiety (-D), depending on whether another part of the linker is present. In this regard, -Y- is bonded to the second spacer unit (-A-) as described herein. The second spacer unit (-A-) can include or be a functional group that can form a bond to another part of the linker (if present) or to the drug moiety (-D). Again, this depends on whether another part of the linker is present. Preferably, the functional group capable of forming a bond to another part of the linker or to the drug moiety (-D) is, for example, It is a carbonyl group depicted as TIFF2024540691000087.tif13128.

[0128] The second spacer unit can be any spacer known to those skilled in the art, for example, a straight or branched chain hydrocarbon-based moiety. The second spacer unit can also include a cyclic moiety, for example, but not limited to, an aromatic moiety. When the second spacer unit is a hydrocarbon-based moiety, the backbone of the second spacer moiety may contain only carbon atoms, but may also contain heteroatoms, such as oxygen (O), nitrogen (N) or sulfur (S) atoms, and / or may contain a carbonyl group (C=O). The second spacer unit can be, for example, (C 1 -C 20 ) a chain of carbon atoms, or (C 1 -C 20) carbon atom chain. In typical embodiments of the hydrocarbon-based second spacer unit, the spacing moiety contains 1 to about 150, 1 to about 100, 1 to about 75, 1 to about 50, or 1 to about 40, or 1 to about 30, or 1 to about 20 (including 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, and 19) main chain atoms. One of skill in the art would know to select a suitable second spacer unit.

[0129] In some embodiments, the second spacer unit (-A-), if present, is 1 -C 10 )Alkylene-C(O)- # , *-(C 3 -C 8 )Carbocyclo-C(O)- # , *-Arylene-C(O)- # , *-(C 1 -C 10 ) Alkylene-arylene-C(O)- # , *-Arylene-(C 1 -C 10 )Alkylene-C(O)- # , *-(C 1 -C 10 )Alkylene-(C 3 -C 8 )Carbocyclo-C(O)- # , *-(C 3 -C 8 )Carbocyclo-(C 1 -C 10 )Alkylene-C(O)- # , *-(C 3 -C 8 )Heterocyclo-C(O)- # , *-(C 1 -C 10 )Alkylene-(C 3 -C 8 )Heterocyclo-C(O)- # , and *-(C 3 -C 8 )Heterocyclo-(C 1 -C 10 )Alkylene-C(O)- #* denotes the point of attachment to -Y-; and # denotes the point of attachment to another part of the linker, if present, or to the drug moiety (-D), depending on whether another part of the linker is present. Preferably, the second spacer unit (-A a -), if present, 3 -C 8 )Carbocyclo-C(O)- # , *-Arylene-C(O)- # , and *-(C 3 -C 8 )Heterocyclo-C(O)- # * denotes the point of attachment to -Y-; and # denotes the point of attachment to another part of the linker, if present, or to the drug moiety (-D), depending on whether another part of the linker is present.

[0130] In other embodiments, the second spacer unit (-A-), if present, is 1 -C 10 ) Alkylene- # , *-(C 3 -C 8 ) Carbocyclo- # , *-Arylene- # , *-(C 1 -C 10 ) Alkylene-arylene- # , *-Arylene-(C 1 -C 10 ) Alkylene- # , *-(C 1 -C 10 )Alkylene-(C 3 -C 8 ) Carbocyclo- # , *-(C 3 -C 8 )Carbocyclo-(C 1 -C 10 ) Alkylene- # , *-(C 3 -C 8 )Heterocyclo- # , *-(C 1 -C 10 )Alkylene-(C 3 -C8 )Heterocyclo- # , and *-(C 3 -C 8 )Heterocyclo-(C 1 -C 10 ) Alkylene- # where * denotes the point of attachment to -Y-; and # denotes the point of attachment to another part of the linker, if present, or to the drug moiety (-D), depending on whether another part of the linker is present. Preferably, the second spacer unit (-A-), if present, is selected from the group consisting of *-(C 3 -C 8 ) Carbocyclo- # , *-Arylene- # , and *-(C 3 -C 8 )Heterocyclo- # * denotes the point of attachment to -Y-; and # denotes the point of attachment to another part of the linker, if present, or to the drug moiety (-D), depending on whether another part of the linker is present or not.

[0131] Preferably, the second spacer unit -A- is TIFF2024540691000088.tif26128, where: TIFF2024540691000089.tif20128 is a five or six membered carbocyclic ring; * indicates the point of attachment to -Y-; and # indicates the point of attachment to another part of the linker, if present, or to the drug moiety (-D), depending on whether another part of the linker is present. The carbocyclic ring can be aromatic or non-aromatic. Preferably, the second spacer unit -A- is TIFF2024540691000090.tif26128, where: TIFF2024540691000091.tif19128 is a five- or six-membered heterocyclic ring containing 1, 2, or 3 heteroatoms independently selected from the group consisting of N, O, and S; * indicates the point of attachment to -Y-; and # indicates the point of attachment to another part of the linker, if present, or to the drug moiety (-D), depending on whether another part of the linker is present. The heterocyclic ring can be aromatic or non-aromatic.

[0132] More preferably, TIFF2024540691000092.tif26128 is TIFF2024540691000093.tif28128, wherein each of A, B, C and D is independently selected from N (nitrogen) and CH; preferably at least one of A, B, C and D is CH; more preferably at least two of A, B, C and D are CH; even more preferably at least three of A, B, C and D are CH, and even more preferably each of A, B, C and D is CH; * indicates a point of attachment to -Y-; and # indicates a point of attachment to another part of the linker, if present, or to the drug moiety (-D), depending on whether another part of the linker is present. Even more preferably, TIFF2024540691000094.tif26128 is TIFF2024540691000095.tif28128, where each of A, B, C and D is independently selected from N (nitrogen) and CH; preferably at least one of A, B, C and D is CH; more preferably at least two of A, B, C and D are CH; even more preferably at least three of A, B, C and D are CH, and even more preferably each of A, B, C and D is CH; where * indicates a point of attachment to -Y-; and # indicates a point of attachment to another part of the linker, if present, or to the drug moiety (-D), depending on whether another part of the linker is present. Even more preferably, TIFF2024540691000096.tif26128 is TIFF2024540691000097.tif25128, where each of A, B, C and D is independently selected from N (nitrogen) and CH; preferably at least one of A, B, C and D is CH; more preferably at least two of A, B, C and D are CH; even more preferably at least three of A, B, C and D are CH, and even more preferably each of A, B, C and D is CH; where * indicates a point of attachment to -Y-; and # indicates a point of attachment to another part of the linker, if present, or to the drug moiety (-D), depending on whether another part of the linker is present. In a highly preferred embodiment, the second spacer unit A is TIFF2024540691000098.tif25128, where * indicates the point of attachment to -Y-; and # indicates the point of attachment to another part of the linker, if present, or to the drug moiety (-D), depending on whether another part of the linker is present.

[0133] In other embodiments, the second spacer unit (-A-) is TIFF2024540691000099.tif19128; and m and n are each, independently, an integer, e.g., 0-20, 0-15, 1-10, 1-8, 1-6, 1-4, 1-3, 1-2, or 1, preferably m is 1 and n is 1; * indicates the position of -Y-, and # indicates the point of attachment, if present, to another part of the linker or to the drug moiety (-D), depending on whether another part of the linker is present or not. Such second spacer units can have, for example, one or two (C 1 -C 8 ) alkyl, optionally substituted.

[0134] Base Z In a preferred embodiment, the second spacer unit -A- is a group Z, where the group Z has the following structure: TIFF2024540691000100.tif26128, where L P is the parallel connector unit; R S are each independently a second polyalkylene glycol unit; M is independently R s and L p is a bond or part that joins with; s* is an integer ranging from 1 to 4; and The wavy line indicates the point of attachment to -Y- and, depending on whether another part of the linker is present, to another part of the linker, if present, or to the drug moiety (-D).

[0135] formula: As shown in TIFF2024540691000101.tif26128, the second polyalkylene glycol unit R S Parallel connector unit L through suitable part M P In some embodiments, M is a bond. In some embodiments, M is a polyalkylene glycol unit linked to a parallel connector unit L P As illustrative examples, M can be any moiety that can be bonded to a cyclic alkyl group, such as -NH-, -O-, S, -C(O)-O-, -C(O)-NH-, and -(C 1 -C 10 ) alkylene. Preferably, each M is independently selected from the group consisting of -NH-, -O-, and -S-. More preferably, each M is -O-.

[0136] The integer s* may range from 1 to 4. Preferably, the integer s* ranges from 1 to 3. More preferably, the integer s* is 1 or 2. Even more preferably, the integer s* is 1. The integer s* is the number of parallel connector units L P The group -MR connected to S Show the number of.

[0137] Parallel Connector Unit (L P ) serves to link -Y- to another part of the linker (L) and, via M, to one or more second polyalkylene glycol units, as indicated by the integer s*. Thus, when present, L P may be any chemical group or moiety capable of linking -Y- to another part of the linker and, via M, to the second polyalkylene glycol unit. Alternatively, a parallel connector unit (L P ) may link Y to the drug moiety (D), if no other parts of the linker are present, and, via M, to the second polyalkylene glycol unit. In this regard, Y may be linked to a parallel connector unit (L P ) are connected to the parallel connector unit (L P ) may include or be a functional group capable of forming a bond to another part of the linker (L) or to the drug moiety (D), depending on whether another part of the linker (L) is present. Preferably, the functional group capable of forming a bond to another part of the linker (L) or to the drug moiety (-D) is, for example, TIFF2024540691000102.tif12128, or a carbonyl group depicted as -C(O)-, or -(C=O)-.

[0138] Parallel Connector Unit (L P ) can be, for example, a straight-chain or branched-chain hydrocarbon-based moiety. P ) can also include a ring portion. P When the parallel connector unit (L) is a hydrocarbon-based moiety, the backbone of the second spacer moiety may contain only carbon atoms, but may also contain heteroatoms such as oxygen (O), nitrogen (N) or sulfur (S) atoms, and / or may contain a carbonyl group (C=O). P ) is, for example, (C 1 -C 20) a chain of carbon atoms, or (C 1 -C 20 ) carbon atom chain. P In typical embodiments, the linking moiety comprises from 1 to about 150, from 1 to about 100, from 1 to about 75, from 1 to about 50, or from 1 to about 40, or from 1 to about 30, or from 1 to about 20 (including 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, and 19) backbone atoms. P The second polyalkylene glycol unit R is connected via M. S Those skilled in the art can easily connect to a suitable parallel connector unit (L P ) to choose.

[0139] In some embodiments, the group Z: TIFF2024540691000103.tif26128, if present, contains 1 to 4, preferably 1 to 3, more preferably 1 or 2, even more preferably 1 group -MR S and each independently represents a substituted *-(C 1 -C 10 )Alkylene-C(O)- # 1 to 4, preferably 1 or 2, more preferably 1 group -MR S and each independently represents a substituted *-(C 3 -C 8 )Carbocyclo-C(O)- # 1 to 4, preferably 1 or 2, more preferably 1 group -MR S each independently represents a substituted *-arylene-C(O)- # 1 to 4, preferably 1 or 2, more preferably 1 group -MR S and each independently represents a substituted *-(C 1 -C 10 ) Alkylene-arylene-C(O)- # 1 to 4, preferably 1 or 2, more preferably 1 group -MR S each independently represents a substituted *-arylene-(C 1 -C 10)Alkylene-C(O)- # 1 to 4, preferably 1 or 2, more preferably 1 group -MR S and each independently represents a substituted *-(C 1 -C 10 )Alkylene-(C 3 -C 8 )Carbocyclo-C(O)- # 1 to 4, preferably 1 or 2, more preferably 1 group -MR S and each independently represents a substituted *-(C 3 -C 8 )Carbocyclo-(C 1 -C 10 )Alkylene-C(O)- # 1 to 4, preferably 1 or 2, more preferably 1 group -MR S and each independently represents a substituted *-(C 3 -C 8 )Heterocyclo-C(O)- # 1 to 4, preferably 1 or 2, more preferably 1 group -MR S and each independently represents a substituted *-(C 1 -C 10 )Alkylene-(C 3 -C 8 )Heterocyclo-C(O)- # and 1 to 4, preferably 1 or 2, more preferably 1 group -MR S and each independently represents a substituted *-(C 3 -C 8 )Heterocyclo-(C 1 -C 10 )Alkylene-C(O)- # * denotes the point of attachment to -Y-; and # denotes the point of attachment to another part of the linker, if present, or to the drug moiety (-D), depending on whether another part of the linker is present. Preferably, the group Z: TIFF2024540691000104.tif26128, if present, contains 1 to 4, preferably 1 or 2, more preferably 1 group -MR S and each independently represents a substituted *-(C 3-C 8 )Carbocyclo-C(O)- # 1 to 4, preferably 1 or 2, more preferably 1 group -MR S each independently represents a substituted *-arylene-C(O)- # and 1 to 4, preferably 1 or 2, more preferably 1 group -MR S and each independently represents a substituted *-(C 3 -C 8 )Heterocyclo-C(O)- # * denotes the point of attachment to -Y-; and # denotes the point of attachment to another part of the linker, if present, or to the drug moiety (-D), depending on whether another part of the linker is present.

[0140] In other embodiments, the group Z: TIFF2024540691000105.tif26128, if present, contains 1 to 4, preferably 1 to 3, more preferably 1 or 2, even more preferably 1 group -MR S and each independently represents a substituted *-(C 1 -C 10 ) Alkylene- # 1 to 4, preferably 1 or 2, more preferably 1 group -MR S and each independently represents a substituted *-(C 3 -C 8 ) Carbocyclo- # 1 to 4, preferably 1 or 2, more preferably 1 group -MR S each independently represents a substituted *-arylene- # 1 to 4, preferably 1 or 2, more preferably 1 group -MR S and each independently represents a substituted *-(C 1 -C 10 ) Alkylene-arylene- # 1 to 4, preferably 1 or 2, more preferably 1 group -MR S each independently represents a substituted *-arylene-(C 1 -C 10 ) Alkylene-# 1 to 4, preferably 1 or 2, more preferably 1 group -MR S and each independently represents a substituted *-(C 1 -C 10 )Alkylene-(C 3 -C 8 ) Carbocyclo- # 1 to 4, preferably 1 or 2, more preferably 1 group -MR S and each independently represents a substituted *-(C 3 -C 8 )Carbocyclo-(C 1 -C 10 ) Alkylene- # 1 to 4, preferably 1 or 2, more preferably 1 group -MR S and each independently represents a substituted *-(C 3 -C 8 )Heterocyclo- # 1 to 4, preferably 1 or 2, more preferably 1 group -MR S and each independently represents a substituted *-(C 1 -C 10 )Alkylene-(C 3 -C 8 )Heterocyclo- # and 1 to 4, preferably 1 or 2, more preferably 1 group -MR S and each independently represents a substituted *-(C 3 -C 8 )Heterocyclo-(C 1 -C 10 ) Alkylene- # * denotes the point of attachment to -Y-; and # denotes the point of attachment to another part of the linker, if present, or to the drug moiety (-D), depending on whether another part of the linker is present. Preferably, the group Z: TIFF2024540691000106.tif26128, if present, contains 1 to 4, preferably 1 or 2, more preferably 1 group -MR S and each independently represents a substituted *-(C 3 -C 8 ) Carbocyclo- #1 to 4, preferably 1 or 2, more preferably 1 group -MR S each independently represents a substituted *-arylene- # and 1 to 4, preferably 1 or 2, more preferably 1 group -MR S and each independently represents a substituted *-(C 3 -C 8 )Heterocyclo- # * denotes the point of attachment to -Y-; and # denotes the point of attachment to another part of the linker, if present, or to the drug moiety (-D), depending on whether another part of the linker is present or not.

[0141] In some embodiments, the group Z: L in TIFF2024540691000107.tif26128 P may be one or more amino acids, which may contain a suitable moiety M, such that a second polyalkylene glycol unit may be attached; preferably s* is 1. The amino acid may be a natural or unnatural amino acid. For example, the amino acid may be selected from the group consisting of lysine, glutamic acid, aspartic acid, serine, tyrosine, threonine, cysteine, selenocysteine, glycine, and homoalanine. In particular, the amino acid may be selected from the group consisting of tyrosine, serine, threonine, glutamic acid, lysine, and glycine. Other suitable moieties L P may be selected from the group consisting of amino alcohols, amino aldehydes and polyamines. Suitable amino acids and further groups for connecting the polyalkylene glycol units are described, for example, in WO 2015 / 057699.

[0142] Preferably, the group Z: TIFF2024540691000108.tif26128 is TIFF2024540691000109.tif34128, where: TIFF2024540691000110.tif19128 is a five- or six-membered carbocyclic ring; the carbocyclic ring can be aromatic or non-aromatic; each M is independently as defined herein; preferably, each M is -O-; R S is each independently a second polyalkylene glycol unit as defined herein; preferably, each R S is independently a second polyethylene glycol unit as defined herein; s* is an integer in the range of 1 to 3, preferably s* is 1 or 2, more preferably s* is 1; * denotes a point of attachment to -Y-; and # denotes a point of attachment to another part of the linker, if present, or to the drug moiety (-D), depending on whether another part of the linker is present.

[0143] More preferably, TIFF2024540691000111.tif34128 is TIFF2024540691000112.tif40165, wherein each of A, B, C and D is CH; R S are each independently a second poly(alkylene) glycol unit as defined herein; preferably, each R S is independently a second polyethylene glycol unit as defined herein; each M is independently as defined herein; preferably each M is -O-; the integer s* is 1 or 2, preferably s* is 1; As shown by TIFF2024540691000113.tif14128, when s* is 2, H is -MR independently in two CHs. S or when s* is 1, in one CH, H is replaced by -MR S * denotes the point of attachment to -Y-; and # denotes the attachment point to another part of the linker, if present (e.g., the amino acid unit -W wEven more preferably, the attachment point to the drug moiety (-D) is indicated. TIFF2024540691000114.tif34128 is TIFF2024540691000115.tif37128, where each of A, B, C and D is C-H; R S are each independently a second poly(alkylene) glycol unit as defined herein; preferably, each R S is independently a second polyethylene glycol unit as defined herein; each M is independently as defined herein; preferably each M is -O-; the integer s* is 1 or 2, preferably s* is 1; As shown by TIFF2024540691000116.tif14128, when s* is 2, H is -MR independently in two CHs. S or when s* is 1, in one CH, H is replaced by -MR S * denotes the point of attachment to -Y-; and # denotes the point of attachment to another part of the linker, if present, or to the drug moiety (-D), depending on whether another part of the linker is present. Even more preferably, TIFF2024540691000117.tif34128 is TIFF2024540691000118.tif34128, where each of A, B, C and D is C-H; R S are each independently a second poly(alkylene) glycol unit as defined herein; preferably, each R S is independently a second polyethylene glycol unit as defined herein; each M is independently as defined herein; preferably each M is -O-; the integer s* is 1 or 2, preferably s* is 1; As shown by TIFF2024540691000119.tif14128, when s* is 2, H is -MR independently in two CHs.S or when s* is 1, in one CH, H is replaced by -MR S * denotes the point of attachment to -Y-; and # denotes the point of attachment to another part of the linker, if present, or to the drug moiety (-D), depending on whether another part of the linker is present. In a highly preferred embodiment, the group Z: TIFF2024540691000120.tif26128 is TIFF2024540691000121.tif27128, where R S is a second polyalkylene glycol unit as described herein; preferably, R S is a second polyethylene glycol unit as defined herein; M is as described herein; preferably M is -O-; * indicates the point of attachment to -Y-; and # indicates the point of attachment to another part of the linker, if present, or to the drug moiety (-D), depending on whether another part of the linker is present.

[0144] In some embodiments, the group Z: TIFF2024540691000122.tif26128 is TIFF2024540691000123.tif36128, where: TIFF2024540691000124.tif20128 is a five- or six-membered heterocyclic ring containing one or two heteroatoms independently selected from the group consisting of N, O, or S; the heterocyclic ring can be aromatic or non-aromatic; each M is independently as defined herein; preferably each M is -O-; R S are each independently a second poly(alkylene) glycol unit as defined herein; preferably, each R Sis independently a second polyethylene glycol unit as defined herein; s* is 1 or 2 (particularly in the case of a six-membered heterocyclic ring), preferably s* is 1 (particularly in the case of a five- or six-membered heterocyclic ring); * indicates the point of attachment to -Y-; and # indicates the point of attachment to another part of the linker, if present, or to the drug moiety (-D), depending on whether another part of the linker is present or not.

[0145] In some embodiments, TIFF2024540691000125.tif37128 is TIFF2024540691000126.tif40161, wherein three of A, B, C and D are independently CH and one of A, B, C and D is independently N; R S are each independently a second poly(alkylene) glycol unit as defined herein; preferably, each R S is independently a second polyethylene glycol unit as defined herein; each M is independently as defined herein; preferably each M is -O-; the integer s* is 1 or 2, preferably s* is 1; As shown by TIFF2024540691000127.tif14128, when s* is 2, H is -MR independently in two CHs. S or when s* is 1, in one CH, H is replaced by -MR S * denotes the point of attachment to -Y-; and # denotes the point of attachment to another part of the linker, if present, or to the drug moiety (-D), depending on whether another part of the linker is present. TIFF2024540691000128.tif35128 is TIFF2024540691000129.tif37128, where three of A, B, C, and D are independently CH, and one of A, B, C, and D is independently N; R S are each independently a second poly(alkylene) glycol unit as defined herein; preferably, each R S is independently a second polyethylene glycol unit as defined herein; each M is independently as defined herein; preferably each M is -O-; the integer s* is 1 or 2, preferably s* is 1; As shown by TIFF2024540691000130.tif14128, when s* is 2, H is -MR independently in two CHs. S or when s* is 1, in one CH, H is replaced by -MR S * denotes the point of attachment to -Y-; and # denotes the point of attachment to another part of the linker, if present, or to the drug moiety (-D), depending on whether another part of the linker is present. TIFF2024540691000131.tif35128 is TIFF2024540691000132.tif34128, where three of A, B, C, and D are independently CH, and one of A, B, C, and D is independently N; R S are each independently a second poly(alkylene) glycol unit as defined herein; preferably, each R S is independently a second polyethylene glycol unit as defined herein; each M is independently as defined herein; preferably each M is -O-; the integer s* is 1 or 2, preferably s* is 1; As shown by TIFF2024540691000133.tif15128, when s* is 2, H is -MR independently in two CHs. Sor when s* is 1, in one CH, H is replaced by -MR S * denotes the point of attachment to -Y-; and # denotes the point of attachment to another part of the linker, if present, or to the drug moiety (-D), depending on whether another part of the linker is present.

[0146] In some embodiments, TIFF2024540691000134.tif35128 is TIFF2024540691000135.tif40164, wherein two of A, B, C and D are independently CH and two of A, B, C and D are independently N; R S are each independently a second poly(alkylene) glycol unit as defined herein; preferably, each R S is independently a second polyethylene glycol unit as defined herein; each M is independently as defined herein; preferably each M is -O-; the integer s* is 1 or 2, preferably s* is 1; As shown by TIFF2024540691000136.tif15128, when s* is 2, H is -MR independently in two CHs. S or, if s* is 1, in one CH, H is replaced by R S * denotes the point of attachment to -Y-; and # denotes the point of attachment to another part of the linker, if present, or to the drug moiety (-D), depending on whether another part of the linker is present. TIFF2024540691000137.tif35128 is TIFF2024540691000138.tif37128, where two of A, B, C, and D are independently CH, and two of A, B, C, and D are independently N; R Sis each independently a second polyalkylene glycol unit as defined herein; preferably, each R S is independently a second polyethylene glycol unit as defined herein; each M is independently as defined herein; preferably each M is -O-; the integer s* is 1 or 2, preferably s* is 1; As shown by TIFF2024540691000139.tif15128, when s* is 2, H is -MR independently in two CHs. S or when s* is 1, in one CH, H is replaced by -MR S * denotes the point of attachment to -Y-; and # denotes the point of attachment to another part of the linker, if present, or to the drug moiety (-D), depending on whether another part of the linker is present. TIFF2024540691000140.tif35128 is TIFF2024540691000141.tif34128, where two of A, B, C, and D are independently CH, and two of A, B, C, and D are independently N; R S are each independently a second poly(alkylene) glycol unit as defined herein; preferably, each R S is independently a second polyethylene glycol unit as defined herein; each M is independently as defined herein; preferably each M is -O-; the integer s* is 1 or 2, preferably s* is 1; As shown by TIFF2024540691000142.tif15128, when s* is 2, H is -MR independently in two CHs. S or when s* is 1, in one CH, H is replaced by -MR S* denotes the point of attachment to -Y-; and # denotes the point of attachment to another part of the linker, if present, or to the drug moiety (-D), depending on whether another part of the linker is present.

[0147] Second polyalkylene glycol unit R S The term “second polyalkylene glycol unit” as used herein refers to a parallel connector unit (L P ) The second polyalkylene glycol unit comprises at least one alkylene glycol subunit. Preferably, the second polyalkylene glycol unit R S has the following structure: More preferably, the second polyalkylene glycol unit R S has the following structure: TIFF2024540691000144.tif15128. Thus, the second polyalkylene glycol unit R S may be a poly(tetramethylene glycol) unit, a poly(propylene glycol) unit, or a poly(ethylene glycol) unit. Even more preferably, the second polyalkylene glycol unit has the following structure: TIFF2024540691000145.tif14128.

[0148] Preferably, the second polyalkylene glycol unit R S Each independently contains 1 to 100 alkylene glycol subunits as described herein. More preferably, the second polyalklyene glycol unit R Seach independently comprises 2 to 50 alkylene glycol subunits. Even more preferably, the second polyalkylene glycol units each independently comprise 3 to 45 alkylene glycol subunits as described herein. Even more preferably, the second polyalkylene glycol units each independently comprise 4 to 40 alkylene glycol subunits as described herein. Even more preferably, the second polyalkylene glycol units each independently comprise 6 to 35 alkylene glycol subunits as described herein. Even more preferably, the second polyalkylene glycol units each independently comprise 8 to 30 alkylene glycol subunits as described herein.

[0149] Preferably, the second polyalkylene glycol unit R S each independently comprises 1 to 20 alkylene glycol subunits as described herein. More preferably, the second polyalkylene glycol units R S each independently comprises 2 to 12 alkylene glycol subunits. Even more preferably, the second polyalkylene glycol units each independently comprise 3 to 11 alkylene glycol subunits as described herein.

[0150] Second polyalkylene glycol unit R S may each independently have the structure: The second polyalkylene glycol unit R may be a polyalkylene glycol unit containing 1 to 100, preferably 2 to 50, more preferably 3 to 45, even more preferably 4 to 40, even more preferably 6 to 35, and still more preferably 8 to 30 subunits having the TIFF2024540691000146.tif15128. S may each independently have the structure: The second polyalkylene glycol unit R may be a polyalkylene glycol unit containing 1 to 100, preferably 2 to 50, more preferably 3 to 45, more preferably 4 to 40, even more preferably 6 to 35, and still more preferably 8 to 30 subunits having the same structure as the first polyalkylene glycol unit R. S may each independently have the structure: In a highly preferred embodiment, the second polyalkylene glycol unit R S may each independently have the structure: It may be a polyethylene glycol unit containing 1 to 100, preferably 2 to 50, more preferably 3 to 45, even more preferably 4 to 40, even more preferably 6 to 35, and still more preferably 8 to 30 subunits having TIFF2024540691000149.tif10128.

[0151] Second polyalkylene glycol unit R S may each independently have the structure: The second polyalkylene glycol unit R S may each independently have the structure: The second polyalkylene glycol unit R S may each independently have the structure: In a highly preferred embodiment, the second polyalkylene glycol unit R S may each independently have the structure: It may be a polyethylene glycol unit containing 1 to 20, preferably 2 to 12, and more preferably 3 to 11 subunits having TIFF2024540691000153.tif10128.

[0152] Preferably, the second polyalkylene glycol unit R S are each independently TIFF2024540691000154.tif15128, Where: TIFF2024540691000155.tif10128 shows the position of M in group Z; K S is H or a second capping group; preferably, K S is -H (hydrogen), -PO 3 H, -(C 1 -C 10 ) alkyl, -(C 1 -C 10 )Alkyl-SO 3 H, -(C 2 -C 10 )Alkyl-CO 2 H, -(C 2 -C 10 ) alkyl-OH, -(C 2 -C 10 )Alkyl-NH 2 , -(C 2 -C 10 )Alkyl-NH(C 1 -C 3 ) alkyl and -(C 2 -C 10 )Alkyl-N((C 1 -C 3 )Alkyl) 2 More preferably, K S is H; and p is an integer ranging from 1 to 100.

[0153] A "second capping group," as referred to herein, may be any moiety capable of functioning as an end group of a second polyalkylene glycol unit. Examples of second capping groups that may be used in the present disclosure include -PO 3 H, -(C 1 -C 10 ) alkyl, -(C 1 -C 10 )Alkyl-SO 3 H, -(C 2 -C 10 )Alkyl-CO 2 H, -(C 2 -C 10 ) alkyl-OH, -(C 2 -C 10 )Alkyl-NH 2 , -(C 2 -C 10 )Alkyl-NH(C 1 -C 3 ) alkyl and -(C 2 -C 10 )Alkyl-N((C 1 -C 3 )Alkyl) 2 In some embodiments, the first capping group is -(C 1 -C 10 ) alkyl, especially methyl.

[0154] Preferably, K S is H (hydrogen).

[0155] The integer p represents the repeat unit in the second polyalkylene glycol unit: TIFF2024540691000156.tif12128 number. Integer p may range from 1 to 100. Preferably, p is in the range from 2 to 50. More preferably, p is in the range from 3 to 45. More preferably, p is in the range from 4 to 40. Even more preferably, p is in the range from 6 to 35. Even more preferably, p is in the range from 8 to 30. Even more preferably, p is in the range from 4 to 16. Even more preferably, p is in the range from 8 to 16. Even more preferably, p is 10, 11, 12, 13 or 14. Even more preferably, p is 11, 12 or 13. In a preferred embodiment, p is 12 or about 12. Even more preferably, p is in the range from 16 to 30. Even more preferably, p is in the range from 20 to 28. Even more preferably, p is 22, 23, 24, 25 or 26. Even more preferably, p is 23, 24 or 25. In a preferred embodiment, p is 24 or about 24. Preferably, the repeat unit is TIFF2024540691000157.tif11128. More preferably, the repeat unit is The file is TIFF2024540691000158.tif12128.

[0156] In the second polyalkylene glycol unit, the integer p can range from 1 to 20. Preferably, p ranges from 2 to 12. More preferably, p ranges from 3 to 11. Preferably, the repeating unit is TIFF2024540691000159.tif11128. More preferably, the repeat unit is The file is TIFF2024540691000160.tif12128.

[0157] Preferably, the second polyalkylene glycol unit R S has the following structure: TIFF2024540691000161.tif10128, i.e., the subunit is designated as an "ethylene glycol subunit." Thus, preferably, the second polyalkylene glycol unit is a second polyethylene glycol unit. The second polyethylene glycol unit comprises at least one ethylene glycol subunit.

[0158] Preferably, the second polyalkylene glycol unit R S may each independently have the structure: The second polyethylene glycol unit may include 1 to 100, preferably 2 to 50, more preferably 3 to 45, more preferably 4 to 40, even more preferably 6 to 35, and still more preferably 8 to 30 subunits having TIFF2024540691000162.tif9128.

[0159] Preferably, the second polyalkylene glycol unit R S may each independently have the structure: The second polyethylene glycol unit may be a second polyethylene glycol unit containing 1 to 20, preferably 2 to 12, and more preferably 3 to 11 subunits having the TIFF2024540691000163.tif9128.

[0160] Preferably, the second polyalkylene glycol unit R S may each independently have the structure: a second polyethylene glycol unit having TIFF2024540691000164.tif15128; Where: TIFF2024540691000165.tif10128 shows the position of M in group Z; K S is H (hydrogen) or a second capping group as described herein; preferably, K S is -H (hydrogen), -PO 3 H, -(C 1 -C 10 ) alkyl, -(C 1-C 10 )Alkyl-SO 3 H, -(C 2 -C 10 )Alkyl-CO 2 H, -(C 2 -C 10 ) alkyl-OH, -(C 2 -C 10 )Alkyl-NH 2 , -(C 2 -C 10 )Alkyl-NH(C 1 -C 3 ) alkyl and -(C 2 -C 10 )Alkyl-N((C 1 -C 3 )Alkyl) 2 More preferably, K S is H; and p is an integer ranging from 1 to 100.

[0161] The integer p represents the repeating unit in the second polyethylene glycol unit: TIFF2024540691000166.tif12128 displays the number. The integer p may be in the range of 1 to 100. Preferably, p is in the range of 2 to 50. More preferably, p is in the range of 3 to 45. Even more preferably, p is in the range of 4 to 40. Even more preferably, p is in the range of 6 to 35. Even more preferably, p is in the range of 8 to 30. Even more preferably, p is in the range of 4 to 16. Even more preferably, p is in the range of 8 to 16. Even more preferably, p is 10, 11, 12, 13 or 14. Even more preferably, p is 11, 12 or 13. In a preferred embodiment, p is 12 or about 12. Even more preferably, p is in the range of 16 to 30. Even more preferably, p is in the range of 20 to 28. Even more preferably, p is 22, 23, 24, 25 or 26. Even more preferably, p is 23, 24 or 25. In a preferred embodiment, p is 24 or about 24.

[0162] In the second polyethylene glycol unit, the integer p can be in the range of 1 to 20. Preferably, p is in the range of 2 to 12. More preferably, p is in the range of 3 to 11.

[0163] Generally, the second polyalkylene glycol unit R S In the second polyethylene glycol unit (preferably the second polyethylene glycol unit), polydisperse polyalkylene glycols (preferably polydisperse polyethylene glycols), monodisperse polyalkylene glycols (preferably monodisperse polyethylene glycols), and individual polyalkylene glycols (preferably individual polyethylene glycols) can be used. Polydisperse polyalkylene glycols (preferably polydisperse polyethylene glycols) are heterogeneous mixtures of sizes and molecular weights, whereas monodisperse polyalkylene glycols (preferably monodisperse polyethylene glycols) are typically purified from heterogeneous mixtures and thus provide a single chain length and molecular weight. The preferred second polyalkylene glycol unit is an individual polyalkylene glycol (preferably individual polyethylene glycol), i.e., a compound that is synthesized stepwise rather than by a polymerization process. An individual polyalkylene glycol (preferably individual polyethylene glycol) provides a single molecule with a defined and specified chain length.

[0164] The second polyalkylene glycol unit (preferably, the second polyethylene glycol unit) provided herein comprises one or more polyalkylene glycol chains (preferably, polyethylene glycol chains). The polyalkylene glycol chains (preferably, polyethylene glycol chains) can be linked together, for example, in a linear, branched or star configuration. Optionally, at least one of the polyalkylene glycol chains (preferably, polyethylene glycol chains) can be derivatized at one end for covalent attachment to M in the group Z.

[0165] The second polyalkylene glycol unit (preferably the second polyethylene glycol unit) is connected to the conjugate (or intermediate thereof) at M in group Z. The other end(s) of the second polyalkylene glycol unit (preferably the second polyethylene glycol unit) is free and untethered and may take the form of hydrogen, methoxy, carboxylic acid, alcohol, or other suitable functional group, such as any second capping group, for example, as described herein. The methoxy, carboxylic acid, alcohol, or other suitable functional group acts as a cap for the terminal polyalkylene glycol subunit (preferably the polyethylene glycol subunit) of the second polyalkylene glycol unit (preferably the second polyethylene glycol unit). Untethered means that the second polyalkylene glycol unit (preferably the second polyethylene glycol unit) is not connected at its untethered site to a drug moiety (D), to a receptor binding molecule, or to a component of a linker (L) that links the drug moiety and / or the receptor binding molecule. For embodiments in which the second polyalkylene glycol unit (preferably, the second polyethylene glycol unit) comprises more than one polyalkylene glycol chain (preferably, the polyethylene glycol chain), the multiple polyalkylene glycol chains (preferably, the polyethylene glycol chains) may be the same or different chemical moieties (e.g., polyalkylene glycols, particularly polyethylene glycols, of different molecular weights or subunit numbers). The multiple second polyalkylene glycol chains (preferably, the second polyethylene glycol chains) are connected to M in the group Z at a single connection site. One of skill in the art will understand that the second polyalkylene glycol unit (preferably, the second polyethylene glycol unit), in addition to comprising repeating polyalkylene glycol subunits (preferably, polyethylene glycol subunits), may further contain non-polyalkylene glycol materials (preferably, non-polyethylene glycol materials) (e.g., to facilitate coupling of the multiple polyalkylene glycol chains (preferably, the polyethylene glycol chains) to each other or to M in the group Z).The non-polyalkylene glycol materials (preferably non-polyethylene glycol materials) are comprised of repeating alkylene glycol subunits (preferably, -CH. 2 CH 2 The term "polyalkylene glycol" refers to an atom in the second polyalkylene glycol unit (preferably, the second polyethylene glycol unit) that is not part of the O-subunit. In embodiments provided herein, the second polyalkylene glycol unit (preferably, the second polyethylene glycol unit) can comprise two monomeric polyalkylene glycol chains (preferably, polyethylene glycol chains) linked to each other via a non-polyalkylene glycol (preferably, non-polyethylene glycol) element. In other embodiments provided herein, the second polyalkylene glycol unit (preferably, the second polyethylene glycol unit) can comprise two linear polyalkylene glycol chains (preferably, polyethylene glycol chains) connected to a central core that is connected to M in the group Z (i.e., the polyalkylene glycol unit (preferably, the polyethylene glycol unit) is branched).

[0166] There are numerous methods of polyalkylene glycol (preferably polyethylene glycol) attachment available to one of skill in the art [see, for example, EP 0 401 384 (PEG coupling to G-CSF); U.S. Pat. No. 5,757,078 (PEGylation of EPO peptides); U.S. Pat. No. 5,672,662 (Polyethylene glycol monosubstituted with propionic or butanoic acid and their functional derivatives for biotechnological applications) and related polymers; U.S. Pat. No. 6,077,939 (PEGylation of the N-terminal α-carbon of peptides); and Veronese (2001) Biomaterials 22:405-417 (review on peptide and protein PEGylation)].

[0167] For example, polyalkylene glycol (preferably polyethylene glycol) can be covalently attached to an amino acid residue via a reactive group. The reactive group is one (e.g., a free amino or carboxyl group) to which an activated polyalkylene glycol molecule (preferably a polyethylene glycol molecule) can be attached. For example, the N-terminal amino acid residue and the lysine (K) residue have a free amino group; the C-terminal amino acid residue has a free carboxyl group. Sulfhydryl groups (e.g., as found in cysteine ​​residues) can also be used as reactive groups to connect polyalkylene glycol (preferably polyethylene glycol). Additionally, enzyme-assisted methods for the specific introduction of activating groups (e.g., hydrazide, aldehyde, and aromatic amino groups) at the C-terminus of polypeptides have been described (see Schwarz, et al. (1990) Methods Enzymol. 184:160; Rose, et al. (1991) Bioconjugate Chem. 2: 154; and Gaertner, et al. (1994) J. Biol. Chem. 269:7224).

[0168] In some embodiments, at least one of the polyalkylene glycol chains (preferably polyethylene glycol chains) making up the second polyalkylene glycol unit (preferably the second polyethylene glycol unit) may be functionalized such that it is functionalized to M in group Z or to a parallel connector unit L in group Z when M is a bond. P The functionalization can be performed, for example, by amine, thiol, NHS ester, alkyne, azide, carbonyl, or other functional groups. The polyalkylene glycol unit (preferably a polyethylene glycol unit) can be coupled to a non-polyalkylene glycol material (preferably a non-polyethylene glycol material, i.e., -CH) to facilitate coupling to M in group Z or to parallel connector units (where M is a bond), or to facilitate coupling of two or more polyalkylene glycol chains (preferably polyethylene glycol chains). 2 CH 2The material may further include a material not consisting of O-.

[0169] In preferred embodiments, the second polyalkylene glycol unit, more preferably the second polyethylene glycol unit, is directly connected to M in group Z. In these embodiments, the second polyalkylene glycol unit, preferably the second polyethylene glycol unit, does not contain a functional group for connection to M in group Z, i.e., M is not connected to a carbon atom of the second polyalkylene glycol unit, more preferably to a CH 3 of the second polyethylene glycol unit. 2 Preferably, in any one of these embodiments, M is not a bond.

[0170] In one group of embodiments, the second polyalkylene glycol unit comprises at least one alkylene glycol subunit, preferably at least two alkylene glycol subunits, more preferably at least three alkylene glycol subunits, even more preferably at least four alkylene glycol subunits, even more preferably at least six alkylene glycol subunits, and even more preferably at least eight alkylene glycol subunits. In some such embodiments, the second polyalkylene glycol unit comprises no more than about 100 alkylene glycol subunits, preferably no more than about 50 alkylene glycol units, more preferably no more than about 45 alkylene glycol subunits, more preferably no more than about 40 alkylene glycol subunits, more preferably no more than about 35 subunits, and even more preferably no more than about 30 alkylene glycol subunits. In any one of these embodiments, the alkylene glycol subunits can be any alkylene glycol subunit as described herein. Preferably, in any one of these embodiments, each alkylene glycol subunit has the following structure: Preferably, when each alkylene glycol subunit is an ethylene glycol subunit, in any one of these embodiments the second polyalkylene glycol unit is a second polyethylene glycol unit.

[0171] In one group of embodiments, the second polyalkylene glycol unit comprises one or more linear polyalkylene glycol chains each having at least one alkylene glycol subunit, preferably at least two alkylene glycol subunits, more preferably at least three alkylene glycol subunits, even more preferably at least four alkylene glycol subunits, even more preferably at least six alkylene glycol subunits, and even more preferably at least eight alkylene glycol subunits. In preferred embodiments, the second polyalkylene glycol unit comprises a total of at least one alkylene glycol subunit, preferably at least two alkylene glycol subunits, more preferably at least three, even more preferably at least four, even more preferably at least six, or even more preferably at least eight alkylene glycol subunits. In some such embodiments, the second polyalkylene glycol unit comprises a total of about 100 or less alkylene glycol subunits, preferably a total of about 50 or less alkylene glycol subunits, more preferably a total of about 45 or less subunits, even more preferably a total of about 40 or less subunits, even more preferably a total of about 35 or less subunits, and even more preferably a total of about 30 or less subunits. In any one of these embodiments, the alkylene glycol subunits can be any alkylene glycol subunit as described herein. Preferably, in any one of these embodiments, each alkylene glycol subunit has the following structure: TIFF2024540691000168.tif10128. Preferably, when each alkylene glycol subunit is an ethylene glycol subunit, in any one of these embodiments the second polyalkylene glycol unit is a second polyethylene glycol unit comprising one or more linear polyethylene glycol chains.

[0172] In another group of embodiments, the second polyalkylene glycol unit comprises a total of 1 to 100, preferably 2 to 50, more preferably 3 to 45, even more preferably 4 to 40, even more preferably 6 to 35, and still more preferably 8 to 30 alkylene glycol subunits. In any one of these embodiments, the alkylene glycol subunits can be any alkylene glycol subunit as described herein. Preferably, in any one of these embodiments, each alkylene glycol subunit has the following structure: Preferably, when each alkylene glycol subunit is an ethylene glycol subunit, in any one of these embodiments the second polyalkylene glycol unit is a second polyethylene glycol unit.

[0173] In another group of embodiments, the second polyalkylene glycol unit comprises one or more linear polyalkylene glycol chains having a total of 1 to 100, preferably 2 to 50, more preferably 3 to 45, even more preferably 4 to 40, even more preferably 6 to 35, and still more preferably 8 to 30 alkylene glycol subunits. In any one of these embodiments, the alkylene glycol subunits can be any alkylene glycol subunit as described herein. Preferably, in any one of these embodiments, each alkylene glycol subunit has the following structure: TIFF2024540691000170.tif10128. Preferably, when each alkylene glycol subunit is an ethylene glycol subunit, in any one of these embodiments the second polyalkylene glycol unit is a second polyethylene glycol unit comprising one or more linear polyethylene glycol chains.

[0174] In another group of embodiments, the second polyalkylene glycol unit is a linear single polyalkylene glycol chain having at least one subunit, preferably at least two subunits, more preferably at least three subunits, even more preferably at least six subunits, and even more preferably at least eight subunits. In any one of these embodiments, the alkylene glycol subunit can be any alkylene glycol subunit as described herein. Preferably, in any one of these embodiments, each alkylene glycol subunit has the following structure: TIFF2024540691000171.tif10128. Preferably, when each alkylene glycol subunit is an ethylene glycol subunit, in any one of these embodiments, the second polyalkylene glycol unit is a second polyethylene glycol unit that is a linear single polyethylene glycol chain. Optionally, in any one of these embodiments, the linear single polyalkylene glycol chain can be derivatized.

[0175] In another group of embodiments, the second polyalkylene glycol unit is a linear single polyalkylene glycol chain having 1 to 100, preferably 2 to 50, more preferably 3 to 45, more preferably 4 to 40, more preferably 6 to 35, more preferably 8 to 30 alkylene glycol subunits. In any one of these embodiments, the alkylene glycol subunits can be any alkylene glycol subunit as described herein. Preferably, in any one of these embodiments, each alkylene glycol subunit has the following structure: TIFF2024540691000172.tif9128. Preferably, when each alkylene glycol subunit is an ethylene glycol subunit, in any one of these embodiments, the second polyalkylene glycol unit is a second polyethylene glycol unit that is a linear single polyethylene glycol chain. Optionally, in any one of these embodiments, the linear single polyalkylene glycol chain can be derivatized.

[0176] In any one of the embodiments provided herein, an exemplary linear polyethylene glycol unit that can be used as the second polyalkylene glycol unit, and in particular as the second polyethylene glycol unit, is as follows: TIFF2024540691000173.tif40128 where the wavy line indicates the site of attachment to M in group Z; R 20 is a PEG connecting unit; preferably, R 20 is absent; more preferably, M is not a bond; R 21 is a PEG capping unit (herein R 21 "K S " is also displayed); R 22 is a PEG coupling unit (i.e., for coupling multiple PEG subunit chains together); n is independently selected from 1 to 100, preferably 2 to 50, more preferably 3 to 45, more preferably 4 to 40, even more preferably 6 to 35, and still more preferably 8 to 30; e is 2 to 5; Each n' is independently selected from 1 to 100, preferably 2 to 50, more preferably 3 to 45, more preferably 4 to 40, even more preferably 6 to 35, and even more preferably 8 to 30. In preferred embodiments, there is at least 1, preferably at least 2, more preferably at least 3, more preferably at least 4, more preferably at least 6, and even more preferably at least 8 ethylene glycol subunits in the polyethylene glycol unit. In some embodiments, there are 100 or less, preferably 50 or less, more preferably 45 or less, more preferably 40 or less, more preferably 35 or less, and even more preferably 30 or less ethylene glycol subunits in the polyethylene glycol unit. R 20 If there is no (CH 2 CH 2 O) The subunit is directly bonded to M in the group Z; more preferably, in such embodiments, M is not a bond.

[0177] Preferably, the linear polyethylene glycol unit is TIFF2024540691000174.tif9128, where the wavy line indicates the site of attachment to M in group Z; 20 , R 21 (As used herein, "K S "), and n are as defined herein; more preferably R 20 is absent; even more preferably, M is not a bond. In a preferred embodiment, n is 12 or about 12. In a preferred embodiment, n is 24 or about 24. Preferably, R 21 is H.

[0178] Polyethylene glycol connecting unit R 20When present, is part of the second polyethylene glycol unit and serves to link the second polyethylene glycol unit to M. In these embodiments, preferably M is not a bond but forms a bond with the second polyethylene glycol unit. In an exemplary embodiment, the PEG connecting unit R 20 If present, *-C(O)- # , *-S(O)- # , *-C(O)O- # , *-C(O)-(C 1 -C 10 )Alkyl- # , *-C(O)-(C 1 -C 10 )Alkyl-O- # , *-C(O)-(C 1 -C 10 )Alkyl-CO 2 - # , *-C(O)-(C 1 -C 10 )Alkyl-NH- # , *-C(O)-(C 1 -C 10 )Alkyl-S- # ;*-C(O)-(C 1 -C 10 )Alkyl-C(O)-NH- # ;*-C(O)-(C 1 -C 10 )Alkyl-NH-C(O)- # ;-(C 1 -C 10 )Alkyl- # , *-(C 1 -C 10 )Alkyl-O- # , *-(C 1 -C 10 )Alkyl-C(O)- # , *-(C 1 -C 10 )Alkyl-C(O)O- # , *-(C 1 -C 10 )Alkyl-NH- # , *-(C 1 -C 10 )Alkyl-S- # , *-(C 1-C 10 )Alkyl-C(O)-NH- # , *-(C 1 -C 10 )Alkyl-NH-C(O)- # , and *-CH 2 -CH 2 SO 2 -(C 1 -C 10 )Alkyl- # , *-CH 2 -C(O)-(C 1 -C 10 )Alkyl- # where * indicates the point of attachment to M in Z and # indicates the point of attachment to the ethylene glycol unit.

[0179] PEG coupling unit R 22 is a part of the second polyethylene glycol unit, if present, and is a repeating -CH 2 CH 2 A non-PEG material that acts to link two or more chains of O-subunits. In an exemplary embodiment, the PEG coupling unit R 22 If present, *-(C 1 -C 10 )Alkyl-C(O)-NH- # , *-(C 1 -C 10 )Alkyl-NH-C(O)- # , *-(C 2 -C 10 )Alkyl-NH- # , *-(C 2 -C 10 )Alkyl-O- # , *-(C 1 -C 10 )Alkyl-S- # , or *-(C 2 -C 10 )Alkyl-NH- # where * denotes a point of attachment to an oxygen atom of the ethylene glycol subunit, and # denotes a point of attachment to a carbon atom of another ethylene glycol subunit.

[0180] As used herein, "K S ", also displayed as the group R 21 is H (hydrogen) in an exemplary embodiment, or may be a second capping group, as described herein; preferably, R 21 -H, -PO 3 H, -(C 1 -C 10 ) alkyl, -(C 1 -C 10 )Alkyl-SO 3 H, -(C 2 -C 10 )Alkyl-CO 2 H, -(C 2 -C 10 ) alkyl-OH, -(C 2 -C 10 )Alkyl-NH 2 , -(C 2 -C 10 )Alkyl-NH(C 1 -C 3 ) alkyl and -(C 2 -C 10 )Alkyl-N((C 1 -C 3 )Alkyl) 2 In some embodiments, R21 is independently selected from the group consisting of -(C 1 -C 10 ) alkyl, particularly methyl. More preferably R 21 is H.

[0181] In any one of the embodiments provided herein, an illustrative linear second polyethylene glycol unit that can be used as the second polyalkylene glycol unit is as follows: TIFF2024540691000175.tif80128, wherein the wavy line indicates the site of attachment to M in group Z; preferably M is not a bond; and each n is 1 to 100, preferably 2 to 50, more preferably 3 to 45, even more preferably 4 to 40, even more preferably 6 to 35, and still more preferably 8 to 30. In some embodiments, n is about 12. In some embodiments, n is about 24.

[0182] In some embodiments, the second polyalkylene glycol unit is about 300 daltons to about 5 kilodaltons; about 300 daltons to about 4 kilodaltons; about 300 daltons to about 3 kilodaltons; about 300 daltons to about 2 kilodaltons; or about 300 daltons to about 1 kilodalton. In some such aspects, the second polyalkylene glycol unit has at least 6 alkylene glycol subunits or at least 8 alkylene glycol subunits. In some such aspects, the second polyalkylene glycol unit may have at least 6 alkylene glycol subunits or at least 8 alkylene glycol subunits, but not more than 100 alkylene glycol subunits, preferably not more than 50 alkylene glycol subunits. In some embodiments, the second polyalkylene glycol unit is a second polyethylene glycol unit that is about 300 daltons to about 5 kilodaltons; about 300 daltons to about 4 kilodaltons; about 300 daltons to about 3 kilodaltons; about 300 daltons to about 2 kilodaltons; or about 300 daltons to about 1 kilodalton. In some such aspects, the second polyethylene glycol unit can have at least 6 ethylene glycol subunits or at least 8 ethylene glycol subunits. In some such aspects, the second polyethylene glycol unit can have at least 6 ethylene glycol subunits or at least 8 ethylene glycol subunits, but not more than 100 ethylene glycol subunits, preferably not more than 50 ethylene glycol subunits.

[0183] In some embodiments, the second polyalkylene glycol unit R S When present, no other alkylene glycol subunits are present in the conjugate of formula (I) (i.e., no alkylene glycol subunits are present in any of the other components of the conjugate, e.g., in another part of the linker L as provided herein). In another aspect, the second polyalkylene glycol unit R S If present, no more than 8, no more than 7, no more than 6, no more than 5, no more than 4, no more than 3, no more than 2 or no more than 1 other alkylene glycol subunits are present in the conjugate of Formula (I) (i.e., no more than 8, 7, 6, 5, 4, 3, 2, or 1 other alkylene glycol subunits are present in other components of the conjugate, such as, for example, in another part of the linker L as provided herein).

[0184] Preferably, in another embodiment, the second polyalkylene glycol unit R S When present, the conjugate comprises R 1 The first polyalkylene glycol unit R F Preferably, R S is the second polyethylene glycol unit and the conjugate is the first polyalkylene glycol unit R F When the first polyalkylene glycol unit further comprises a first polyethylene glycol unit, as described herein.

[0185] It will be understood that in referring to alkylene glycol subunits, particularly ethylene glycol subunits, and depending on the context, the number of subunits may represent an average number, for example, when referring to a population of conjugates or intermediate compounds and using polydisperse polyalkylene glycols, particularly polydisperse polyethylene glycols.

[0186] Linker*-A a -W w-B b - ## In some embodiments, the linker L has the formula: a -W w -B b - ## where: -A- is a second spacer unit, as described herein; a is 0 or 1; each -W- is independently an amino acid; w is independently an integer ranging from 0 to 12; -B- is a first spacer unit; and b is 0 or 1; * denotes a point of attachment to -Y-; and ## denotes a point of attachment to the drug moiety. As used herein, the notation "W w ", or "-W w - ", i.e. the combination of W and the associated integer w is also designated an "amino acid unit". Examples for suitable second spacer units, amino acid units and first spacer units are described, for example, in WO 2004 / 010957 A2.

[0187] Structure*-A a -W w -B b - ## In a linker having the formula: w -. The second spacer unit (-A-) can be any second spacer unit as described herein. When present, the second spacer unit (-A-) can be any chemical group or moiety capable of linking -Y- to an amino acid unit. Alternatively, the second spacer unit can link -Y- to the first spacer unit when the amino acid unit is not present. Alternatively, the second spacer unit can link -Y- to the drug moiety (-D) when the first spacer unit and the amino acid unit are not present. In this regard, -Y- is attached to the second spacer unit (-A-) as described herein. The second spacer unit (-A-) can be any chemical group or moiety capable of linking -Y- to an amino acid unit (-W). w -) and / or the first spacer unit (-B-) are present, w-) or to the first spacer unit (-B b Preferably, the amino acid unit (-W) may comprise or be a functional group capable of forming a bond to the amino acid unit (-W) or to a drug moiety (-D). w Functional groups capable of forming a bond to the N-terminus of an amino acid unit (-B-), in particular to the N-terminus of an amino acid unit, or to the first spacer unit (-B-), or to the d moiety (-D) are, for example, TIFF2024540691000176.tif11128 or a carbonyl group depicted as -C(O)-. The integer a associated with the second spacer unit can be 0 or 1. Preferably, the integer a is 1. Alternatively, in other embodiments, the second spacer unit is absent (a = 0).

[0188] Linker*-A a -W w -B b - ## In the formula (I), the second spacer unit -A-, when present, can be any second spacer unit as described herein. a -W w -B b - ## In a preferred embodiment, the second spacer unit -A-, if present (a=1), has the structure: TIFF2024540691000177.tif26128, where TIFF2024540691000178.tif26128 is as defined herein. Thus, in a preferred embodiment, the linker (L) has the structure: TIFF2024540691000179.tif29128, where L P , R S , s*, M, W, w, B and b are as defined herein; * denotes the point of attachment to -Y-; and ## denotes the point of attachment to the drug moiety (-D).

[0189] Amino acid unit (-W w-), when present, may link the second spacer unit A to the first spacer unit B if the first spacer unit is present. Alternatively, the amino acid unit may link the second spacer unit to the drug moiety (D) if the first spacer unit is not present. Alternatively, the amino acid unit may link Y to the first spacer unit if the second spacer unit is not present. Alternatively, the amino acid unit may link Y to the drug moiety if the first spacer unit and the second spacer unit are not present.

[0190] Amino acid unit-W w - may be a dipeptide (w = 2), tripeptide (w = 3), tetrapeptide (w = 4), pentapeptide (w = 5), hexapeptide (w = 6), heptapeptide (w = 7), octapeptide (w = 8), nonapeptide (w = 9), decapeptide (w = 10), undecapeptide (w = 11) or dodecapeptide (w = 12).

[0191] In some embodiments, the amino acid unit can comprise a naturally occurring amino acid. In some embodiments, the amino acid unit can comprise a non-naturally occurring amino acid.

[0192] In any one of the embodiments described herein, each amino acid of the amino acid unit may be independently in the L-configuration or the D-configuration, except for non-chiral amino acids such as, for example, glycine. Preferably, in any one of the embodiments described herein, each amino acid of the amino acid unit is in the L-configuration (i.e., the naturally occurring configuration), except for non-chiral amino acids such as, for example, glycine.

[0193] Preferably, when the second spacer unit (-A-) is present, in any one of the embodiments described herein, the amino acid unit -W w The N-terminus of - is more preferably linked to the second spacer unit (A) via a carbonyl group of the second spacer unit. Preferably, in any one of the embodiments described herein, the amino acid unit -W wThe C-terminus of - is linked to the first spacer unit (B), if the first spacer unit is present. Alternatively, in any one of the embodiments described herein, the amino acid unit -W w The C-terminus of - can be attached to a drug moiety (-D) when the first spacer unit is absent. w The N-terminus of - may be attached to the first spacer unit (B), if present, and the C-terminus may be attached to the second spacer unit A, if present.

[0194] Each -W- unit may independently have the formula depicted in square brackets below, where w is an integer ranging from 0 to 12; preferably w is an integer ranging from 1 to 5; more preferably w is an integer ranging from 2 to 4; even more preferably w is 2 or 3; in a highly preferred embodiment, w is 2: TIFF2024540691000180.tif28128 where R 19 is hydrogen, methyl, isopropyl, isobutyl, sec-butyl, benzyl, p-hydroxybenzyl, -CH 2 OH, -CH(OH)CH 3 , -CH 2 CH 2 SCH 3 , -CH 2 CONH 2 , -CH 2 COOH, -CH 2 CH 2 CONH 2 , -CH 2 CH 2 COOH, -(CH 2 ) 3 NHC(=NH)NH 2 , -(CH 2 ) 3 NH 2 , -(CH 2 ) 3 NHCOCH 3 , -(CH 2 ) 3 NHCHO, -(CH 2 ) 4 NHC(=NH)NH 2 , -(CH 2 )4 NH 2 , -(CH 2 ) 4 NHCOCH 3 , -(CH 2 ) 4 NHCHO, -(CH 2 ) 3 NHCONH 2 , -(CH 2 ) 4 NHCONH 2 , -CH 2 CH 2 CH(OH)CH 2 NH 2 , 2-pyridylmethyl-, 3-pyridylmethyl-, 4-pyridylmethyl-, phenyl, cyclohexyl, The file is TIFF2024540691000181.tif94150.

[0195] The amino acid unit can be enzymatically cleaved by one or more enzymes, including but not limited to a tumor-associated protease, preferably a cathepsin, more preferably cathepsin B, to liberate the drug moiety (-D), which in one embodiment is protonated in vivo upon release to provide the free drug moiety (D). w The - unit is represented by formulas (VII) to (IX).

[0196] Therefore, -W w The -unit may be a dipeptide of formula (VII): TIFF2024540691000182.tif23128 in formula, R 20 and R 21 is as follows: TIFF2024540691000183.tif64128.

[0197] -W w The -unit may be a tripeptide of formula (VIII): TIFF2024540691000184.tif23128 formula, R 20 , R 21 and R 22 is as follows: TIFF2024540691000185.tif23128.

[0198] W w The unit may be a tripeptide of formula (IX): TIFF2024540691000186.tif25128 in formula, R 20 , R 21 , R 22 and R 23 is as follows: TIFF2024540691000187.tif21128.

[0199] Exemplary amino acid units include R 20 is benzyl and R 21 Ga-(CH 2 ) 4 NH 2 (Phe-Lys);R 20 is isopropyl and R 21 Ga-(CH 2 ) 4 NH 2 (Val-Lys);R 20 is isopropyl and R 21 Ga-(CH 2 ) 3 NHCONH 2 Another exemplary amino acid unit includes, but is not limited to, a unit of formula (VII) where R 20 is benzyl and R 21 is benzyl and R 22 Ga-(CH 2 ) 4 NH 2 (Phe-Phe-Lys), a unit of formula (VIII).

[0200] Useful -W w The -W units can be designed and optimized in their selectivity for enzymatic cleavage by a particular enzyme, e.g., a tumor-associated protease. wThe -W unit is one whose cleavage is catalyzed by cathepsin B, C and / or D, or plasmin protease ("tumor-associated protease"). w - The unit is cleaved by cathepsin B. Suitable linkers that can be cleaved by a protease are described, for example, in GM Dubowchik et al., "Cathepsin B-Labile Dipeptide Linkers for Lysosomal Release of Doxorubicin from Internalizing Immunoconjugates; Model Studies of Enzymatic Drug Release and Antigen-Specific In Vitro Anticancer Activity", Bioconjugate Chem., Vol. 13, No. 4, 2002, 855-869; SC Jeffrey et al., "Dipeptide-based highly potent doxorubicin antibody conjugate", Bioorg. Med. Chem. Lett. 16 (2006), 358-362; and MS Kung Sutherland et al., "SGN-CD33A: a novel CD33-targeting antibody-drug conjugate using a pyrrolobenzodiazepine dimer is active in models of drug-resistant AML", Blood, 22 August 2013, volume 11, pages 117-119. 122, number 8, 1455-1463.

[0201] In one embodiment, -W w - is a dipeptide, tripeptide, tetrapeptide or pentapeptide. w - is a dipeptide, tripeptide or tetrapeptide. More preferably, -W w - is a dipeptide or a tripeptide. In a highly preferred embodiment, -W w- is a dipeptide (i.e. w = 2).

[0202] R 19 , R 20 , R 21 , R 22 or R 23 If is other than hydrogen, R 19 , R 20 , R 21 , R 22 or R 23 The carbon atom to which R is attached is chiral. 19 , R 20 , R 21 , R 22 or R 23 Each carbon atom to which R is attached may independently be in the (S) or (R) configuration. 19 , R 20 , R 21 , R 22 or R 23 Each carbon atom to which is attached is in the (S) configuration.

[0203] In one preferred embodiment, the amino acid unit is valine-citrulline (i.e., Val-Cit or VC). In another preferred embodiment, the amino acid unit is valine-alanine (i.e., Val-Ala or VA). In another preferred embodiment, the amino acid unit is alanine-alanine (i.e., Ala-Ala or AA). In another preferred embodiment, the amino acid unit is phenylalanine-lysine (i.e., Phe-Lys or FK). Such linkers are illustrative examples of linkers that can be cleaved by a protease, such as, for example, cathepsin B.

[0204] The designation of peptides used throughout the specification follows conventional nomenclature. Thus, the N-terminus of the peptide is written on the left and the C-terminus of the peptide is written on the right. As an illustrative but non-limiting example, in the dipeptide valine-citrulline (i.e. Val-Cit or VC), valine is the N-terminus and citrulline is the C-terminus. Preferably, in any one of the embodiments described herein, if the second spacer unit (-A-) is present, the N-terminus of the peptide, such as a dipeptide (as an illustrative but non-limiting example: Val-Cit), is linked to the second spacer unit (-A-), more preferably via the carbonyl group of the second spacer unit, and the C-terminus of the peptide is linked to the first spacer unit (-B-), if the first spacer unit (-B-) is present, or to the drug moiety (-D) if the first spacer unit (-B-) is absent.

[0205] In yet another embodiment, the amino acid unit is N-methylvaline-citrulline. In yet another embodiment, the amino acid unit is selected from the group consisting of 5-aminovaleric acid, homophenylalanine-lysine, tetraisoquinolinecarboxylate-lysine, cyclohexylalanine-lysine, isonepecotic acid-lysine, beta-alanine-lysine, glycine-serine-valine-glutamine, and isonepecotic acid.

[0206] Preferably, the amino acid unit is a dipeptide selected from the group consisting of valine-citrulline (i.e. Val-Cit or VC), valine-alanine (i.e. Val-Ala or VA), alanine-alanine (i.e. Ala-Ala or AA) and phenylalanine-lysine (i.e. Phe-Lys or FK). More preferably, the amino acid unit is a dipeptide selected from the group consisting of valine-citrulline (i.e. Val-Cit or VC), valine-alanine (i.e. Val-Ala or VA), and phenylalanine-lysine (i.e. Phe-Lys or FK). Even more preferably, the amino acid unit is valine-citrulline (i.e. Val-Cit or VC) or valine-alanine (i.e. Val-Ala or VA). Even more preferably, the amino acid unit is valine-citrulline (i.e. Val-Cit or VC).

[0207] In some embodiments, the amino acid unit is selected from the group consisting of valine-glutamine (i.e., Val-Gln or VQ), leucine-glutamine (i.e., Leu-Gln or LQ), phenylalanine-glutamine (i.e., Phe-Gln or FQ) and threonine-threonine (i.e., Thr-Thr or TT). Preferably, the amino acid unit is selected from the group consisting of valine-glutamine (i.e., Val-Gln or VQ), leucine-glutamine (i.e., Leu-Gln or LQ), and phenylalanine-glutamine (i.e., Phe-Gln or FQ). More preferably, the amino acid unit is valine-glutamine (i.e., Val-Gln or VQ) or leucine-glutamine (i.e., Leu-Gln or LQ). Linkers comprising amino acid units according to these embodiments may be illustrative examples for linkers that are cleavable, particularly by proteases such as cathepsins (e.g., cathepsin B). These embodiments of the amino acid unit and further suitable amino acid units are disclosed, for example, in Salomon et al., "Optimizing Lysosomal Activation of Antibody-Drug Conjugates (ADCs) by Incorporation of Novel Cleavable Dipeptide Linkers", Mol. Pharmaceutics 2019, 16, 12, 4817-4825.

[0208] The first spacer unit (-B-), if present, is spaced from the amino acid unit (W w ) to the drug moiety. Alternatively, the first spacer unit (B) may link the second spacer unit (A) to the drug moiety (C) when the amino acid unit is absent. The first spacer unit may link the drug moiety to Y when both the amino acid unit and the second spacer unit are absent.

[0209] The integer b can be 0 or 1. In preferred embodiments, the integer b is 1. Alternatively, in other embodiments, the integer b is 0 and the first spacer unit is absent.

[0210] The first spacer unit (-B-) can be of two general types: self-immolative and non-self-immolative. A non-self-immolative first spacer unit is one in which some or all of the first spacer unit is an amino acid unit (-W w -) that remains attached to the drug moiety (D). Examples of non-self-immolative first spacer units include, but are not limited to, (glycine-glycine) first spacer units and glycine first spacer units (both depicted in Scheme 1) (below). When an exemplary compound containing a glycine-glycine first spacer unit or a glycine first spacer unit undergoes enzymatic cleavage via a tumor cell-associated protease, a cancer cell-associated protease or a lymphocyte-associated protease, the glycine-glycine-drug moiety (where "D" represents a drug moiety) or glycine-drug moiety (D) is converted to -A. a -W w - In one embodiment, an independent hydrolysis reaction occurs within the target cell, cleaving the glycine-drug moiety bond and liberating the drug (D). TIFF2024540691000188.tif53128

[0211] In one embodiment, the non-self-immolative first spacer unit is -Gly-Gly-. In another embodiment, the non-self-immolative first spacer unit is -Gly-.

[0212] Alternatively, exemplary compounds containing a self-immolative first spacer unit can release the drug moiety-D without the need for a separate hydrolysis step. In an exemplary embodiment, the self-immolative first spacer unit is a PAB group linked to -Ww- through the amino nitrogen atom of the PAB group and directly linked to -D through a carbonate, carbamate or ether group. Without being bound by any particular theory or mechanism, Scheme 2 depicts a possible mechanism of drug release of a PAB group directly connected to -D through a carbamate or carbonate group, as supported by Toki et al. (2002) J Org. Chem. 67:1866-1872. TIFF2024540691000189.tif102128Here, Q is -(C 1 -C 8 ) alkyl, -O-(C 1 -C 8 ) alkyl, -halogen, -nitro or -cyano; m is an integer ranging from 0 to 4, preferably m is 0, 1 or 2, more preferably m is 0 or 1, even more preferably m is 0; and p is in the range of 1 to 20.

[0213] Without being bound by any particular theory or mechanism, Scheme 3 depicts a possible mechanism of drug release for a PAB group that is directly connected to the drug moiety-D via an ether or amine bond. TIFF2024540691000190.tif110128Here, Q is -(C 1 -C 8 ) alkyl, -O-(C 1 -C 8 ) alkyl, -halogen, -nitro or -cyano; m is an integer ranging from 0 to 4, preferably m is 0, 1 or 2, more preferably m is 0 or 1, even more preferably m is 0; and p is in the range of 1 to 20.

[0214] Other examples of self-immolative spacers include, but are not limited to, aromatic compounds that are electronically similar to the PAB group, such as 2-aminoimidazole-5-methanol derivatives (Hay et al. (1999) Bioorg. Med. Chem. Lett. 9:2237) and ortho- or para-aminobenzyl acetals. Spacers that undergo cyclization upon amide bond hydrolysis can be used, such as substituted and unsubstituted 4-aminobutyric acid amides (Rodrigues et al., Chemistry Biology, 1995, 2, 223), appropriately substituted bicyclo[2.2.1] and bicyclo[2.2.2] ring systems (Storm, et al., J. Amer. Chem. Soc., 1972, 94, 5815), and 2-aminophenylpropionic acid amides (Amsberry, et al., J. Org. Chem., 1990, 55, 5867). Elimination of amine-containing drugs substituted at the alpha position of glycine (Kingsbury, et al., J. Med. Chem., 1984, 27, 1447) is also an example of a self-immolative spacer useful in the exemplary compounds.

[0215] In one embodiment, the first spacer unit is a branched bis(hydroxymethyl)styrene (BHMS) unit as depicted in Scheme 4, which can be used to incorporate and release multiple drugs (D). TIFF2024540691000191.tif44128Here, Q is -(C 1 -C 8 ) alkyl, -O-(C 1 -C 8 ) alkyl, -halogen, -nitro or -cyano; m is an integer ranging from 0 to 4; preferably m is 0, 1 or 2; more preferably m is 0 or 1; even more preferably m is 0; and p is in the range of 1 to 10; n is 0 or 1; and p is in the range of 1 to 20.

[0216] In a preferred embodiment, the first spacer unit has the formula (X): TIFF2024540691000192.tif27128, where Q is -(C 1 -C 8 ) alkyl, -O-(C 1 -C 8 ) alkyl, -halogen, -nitro or -cyano; and m is an integer ranging from 0 to 4; preferably m is 0, 1 or 2; more preferably m is 0 or 1; in a highly preferred embodiment m is 0.

[0217] In some embodiments, the first spacer unit has formula (XI): Represented by TIFF2024540691000193.tif9128.

[0218] In some embodiments, the first spacer unit has formula (XII): Represented by TIFF2024540691000194.tif15128.

[0219] Preferably, in any one of formulas (X), (XI) and (XII), particularly in formula (X), when an amino acid unit is present, the NH group is attached to the C-terminus of the amino acid unit. Preferably, in any one of formulas (X), (XI) and (XII), particularly in formula (X), the C(O) group is attached to the drug moiety (D).

[0220] In a highly preferred embodiment, the first spacer unit has the following structure: Preferably, when an amino acid unit is present, the NH group is a PAB group having the amino acid unit (-W w -), more preferably to the C-terminus of the amino acid unit. Preferably, the C(O) group is attached to the drug moiety (D).

[0221] In some embodiments, the first spacer group (-B-) is a heterocyclic "self-immolative moiety" of Formula I, II or III attached to the drug moiety and incorporates an amide group which, upon hydrolysis by an intracellular protease, initiates a reaction that ultimately cleaves the first spacer unit (-B-) from the drug moiety such that the drug is released from the conjugate in an active form. The linker moiety comprises an amino acid unit (-W) adjacent to the first spacer group (-B-) that is a substrate for an intracellular enzyme, e.g., an intracellular protease such as a cathepsin (e.g., cathepsin B), which cleaves the peptide at the amide bond shared with the first spacer group (-B-). w Heterocyclic self-immolative moieties are described, for example, in WO 2019 / 236954.

[0222] In some embodiments, the first spacer unit (-B-) is a heterocyclic self-immolative group selected from formulas I, II, and III: TIFF2024540691000196.tif77128In the formula, the wavy line represents the amino acid unit -W w - and indicates the site of covalent attachment to the drug moiety, where U is O, S or NR 6 and Q is CR 4 or N;V 1 , V 2 and V 3 is an independent CR 4 or N, with the proviso that for formulas II and III, Q, V 1 and V 2 at least one of is N; T is O pending from a drug moiety (-D); R 1 , R 2 , R 3 and R 4 are H, F, Cl, Br, I, OH, -N(R 5 ) 2 , -N(R 5 ) 3 + , -(C 1 -C 8 ) Alkyl halides, carboxylates, sulfates, sulfamates, sulfonates, -SO 2 R5 , -S(=O)R 5 , -SR 5 , -SO 2 N(R 5 ) 2 , -C(=O)R 5 , -CO 2 R 5 , -C(=O)N(R 5 ) 2 , -CN, -N 3 , -NO 2 , -(C 1 -C 8 )alkoxy, -(C 1 -C 8 ) halo-substituted alkyl, polyethyleneoxy, phosphonate, phosphate, -(C 1 -C 8 ) alkyl, -(C 1 -C 8 ) substituted alkyl, -(C 2 -C 8 ) alkenyl, -(C 2 -C 8 ) substituted alkenyl, -(C 2 -C 8 ) alkynyl, -(C 2 -C 8 ) substituted alkynyl, -(C 6 -C 20 )aryl, -(C 6 -C 20 ) substituted aryl, -(C 3 -C 20 ) heterocycles, and -(C 3 -C 20 ) substituted heterocycles; or when taken together, R 2 and R 3 forms a carbonyl (=O) or a spirocarbocyclic ring of 3 to 7 carbon atoms; and R 5 and R 6 is H, -(C 1 -C 8 ) alkyl, -(C 1 -C 8 ) substituted alkyl, -(C 2 -C 8 ) alkenyl, -(C 2 -C 8 ) substituted alkenyl, -(C2 -C 8 ) alkynyl, -(C 2 -C 8 ) substituted alkynyl, -(C 6 -C 20 )aryl, -(C 6 -C 20 ) substituted aryl, -(C 3 -C 20 ) heterocycles, and -(C 3 -C 20 ) substituted heterocycles; 1 -C 8 ) substituted alkyl, -(C 2 -C 8 ) substituted alkenyl, -(C 2 -C 8 ) substituted alkynyl, -(C 6 -C 20 ) substituted aryl, and -(C 3 -C 20 ) Substituted heterocycles are F, Cl, Br, I, OH, -N(R 5 ) 2 , -N(R 5 ) 3 + , -(C 1 -C 8 ) alkyl halides, carboxylates, sulfates, sulfamates, sulfonates, -(C 1 -C 8 ) alkyl sulfonates, -(C 1 -C 8 ) alkylamino, 4-dialkylaminopyridinium, -(C 1 -C 8 ) alkyl hydroxyl, -(C 1 -C 8 ) Alkylthiol, -SO 2 R 5 , -S(=O)R 5 , -SR 5 , -SO 2 N(R 5 ) 2 , -C(=O)R 5 , -CO 2 R 5 , -C(=O)N(R 5 ) 2 , -CN, -N3 , -NO 2 , -(C 1 -C 8 )alkoxy, -(C 1 -C 8 )trifluoroalkyl, -(C 1 -C 8 ) alkyl, -(C 3 -C 12 ) carbocycle, -(C 6 -C 20 )aryl, -(C 3 -C 20 ) is independently substituted with one or more substituents selected from the group consisting of heterocycle, polyethyleneoxy, phosphonate, and phosphate.

[0223] Conjugates that include heterocyclic self-immolative moieties are stable extracellularly or in the absence of enzymes capable of cleaving the amide bond of the self-immolative moiety, but upon entry into a cell or exposure to a suitable enzyme, the amide bond is cleaved, initiating a spontaneous self-immolation reaction that cleaves the bond covalently linking the self-immolative moiety to the drug moiety, thereby resulting in release of the drug in its underivatized or pharmacologically active form.

[0224] The self-immolative moiety in the conjugate may incorporate one or more heteroatoms, thereby providing improved solubility, improving cleavage rates, and / or reducing the aggregation tendency of the conjugate. Thus, heterocyclic self-immolative linker constructs may in some cases result in increased efficacy, reduced toxicity, and / or desirable pharmacokinetic and / or pharmacodynamic properties.

[0225] T in formulas I-III is understood to be O because it is derived from a tertiary hydroxyl (-OH) on the lactone ring portion of the drug moiety.

[0226] Without being limited by theory or by any particular mechanism, the presence of electron-withdrawing groups on the heterocyclic ring of Formula I, II, or III may slow the rate of cleavage.

[0227] In one embodiment, the self-immolative moiety is a group of formula I, where Q is N and U is O or S. Such groups have non-linear structural features that improve the solubility of the conjugate. In this context, R is sometimes H, methyl, nitro, or CF 3 In one embodiment, Q is N and U is O, thereby forming an oxazole ring, and R is H. In another embodiment, Q is N and U is S, thereby forming Me or CF at R. 3 The thiazole ring may be optionally substituted with a group.

[0228] In another exemplary embodiment, the self-immolative moiety is 1 and V 2 is a group of formula II, wherein Q, V are independently N or CH. 1 and V 2 Each of Q and V is N. 1 is N, while V 2 In another embodiment, Q and V are 2 is N, while V 1 In another embodiment, Q and V are 1 are both CH and V 2 is N. In another embodiment, Q is N, while V 1 and V 2 are both CH.

[0229] In another embodiment, the self-immolative moiety is Q, V 1 , V 2 and V 3 is a group of formula III, wherein each is independently N or CH. In another embodiment, Q is N, while V 1 , V 2 and V 3 Each of Q, V is N. 1 , and V 2 are CH, while V 3 is N. In another embodiment, Q, V2 and V 3 are CH, while V 1 is N. In another embodiment, Q, V 1 and V 3 are CH, while V 2 is N. In another embodiment, Q and V 2 are both N, while V 1 and V 3 In another embodiment, Q and V are both CH. 2 are both CH, while V 1 and V 3 are both N. In another embodiment, Q and V 3 are both N, while V 1 and V 2 are both CH.

[0230] Preferably, the linker (L) has the formula: a -W w -B b - ## wherein integer a is 1, integer b is 1, and integer w is 2, 3 or 4, more preferably integer w is 2 or 3; in a highly preferred embodiment, integer w is 2; and -A-, each -W- and -B- are as defined herein; * indicates a point of attachment to Y; and ## indicates a point of attachment to the drug moiety (D).

[0231] Preferably, the linker (L) has the following structure: *-A a -W w -B b - ## where -A- is a second spacer unit as described herein; a is an integer as described herein; preferably a is 1; -B- is a first spacer unit as described herein; b is an integer as described herein; preferably b is 1; * denotes the point of attachment to the Y; and ## denotes the point of attachment to the drug moiety (-D); -W w - is a dipeptide selected from the group consisting of valine-citrulline (i.e. Val-Cit or VC), valine-alanine (i.e. Val-Ala or VA), alanine-alanine (i.e. Ala-Ala or AA) and phenylalanine-lysine (i.e. Phe-Lys or FK). Preferably, in these embodiments, the amino acid unit is a dipeptide selected from the group consisting of valine-citrulline (i.e. Val-Cit or VC), valine-alanine (i.e. Val-Ala or VA), and phenylalanine-lysine (i.e. Phe-Lys or FK). Even more preferably, in these embodiments, the amino acid unit is valine-citrulline (i.e. Val-Cit or VC) or valine-alanine (i.e. Val-Ala or VA). Even more preferably, in these embodiments, the amino acid unit is valine-citrulline (i.e. Val-Cit or VC). Alternatively, in these embodiments, the amino acid unit -W w - may be a dipeptide selected from the group consisting of valine-glutamine (i.e. Val-Gln or VQ), leucine-glutamine (i.e. Leu-Gln or LQ), phenylalanine-glutamine (i.e. Phe-Gln or FQ) and threonine-threonine (i.e. Thr-Thr or TT). In these embodiments, the amino acid unit may be a dipeptide selected from the group consisting of valine-glutamine (i.e. Val-Gln or VQ), leucine-glutamine (i.e. Leu-Gln or LQ), and phenylalanine-glutamine (i.e. Phe-Gln or FQ). In these embodiments, the amino acid unit may be valine-glutamine (i.e. Val-Gln or VQ) or leucine-glutamine (i.e. Leu-Gln or LQ). Linkers according to these embodiments may be illustrative examples for linkers that are cleavable, inter alia, by a protease, such as, for example, a cathepsin (e.g., cathepsin B).

[0232] Preferably, the linker L has the structure: TIFF2024540691000197.tif31128, where -Aa - is a second spacer unit as described herein; a is an integer as described herein; preferably a is 1; -W w - is an amino acid unit as described herein; w is an integer as described herein; preferably, w is 2, 3 or 4 (i.e., preferably, -W w - is a dipeptide, tripeptide or tetrapeptide), and more preferably w is 2 or 3 (i.e., more preferably -W w - is a dipeptide or tripeptide), and in highly preferred embodiments, w is 2 (i.e., even more preferably, -W w - is a dipeptide); Q is as defined herein; m is an integer as defined herein, preferably m is 0; * denotes the point of attachment to Y; and ## denotes the point of attachment to the drug moiety (-D). Preferably, in these embodiments, the amino acid unit -W w - is a dipeptide selected from the group consisting of valine-citrulline (i.e. Val-Cit or VC), valine-alanine (i.e. Val-Ala or VA), alanine-alanine (i.e. Ala-Ala or AA) and phenylalanine-lysine (i.e. Phe-Lys or FK). More preferably, in these embodiments, the amino acid unit is a dipeptide selected from the group consisting of valine-citrulline (i.e. Val-Cit or VC), valine-alanine (i.e. Val-Ala or VA), and phenylalanine-lysine (i.e. Phe-Lys or FK). Even more preferably, in these embodiments, the amino acid unit is valine-citrulline (i.e. Val-Cit or VC) or valine-alanine (i.e. Val-Ala or VA). Even more preferably, in these embodiments, the amino acid unit is valine-citrulline (i.e. Val-Cit or VC). Alternatively, in these embodiments, the amino acid unit -W w- may be a dipeptide selected from the group consisting of valine-glutamine (i.e. Val-Gln or VQ), leucine-glutamine (i.e. Leu-Gln or LQ), phenylalanine-glutamine (i.e. Phe-Gln or FQ) and threonine-threonine (i.e. Thr-Thr or TT). In these embodiments, the amino acid unit may be a dipeptide selected from the group consisting of valine-glutamine (i.e. Val-Gln or VQ), leucine-glutamine (i.e. Leu-Gln or LQ), and phenylalanine-glutamine (i.e. Phe-Gln or FQ). In these embodiments, the amino acid unit may be valine-glutamine (i.e. Val-Gln or VQ) or leucine-glutamine (i.e. Leu-Gln or LQ). Linkers according to these embodiments may be illustrative examples for linkers that are cleavable, inter alia, by a protease, such as, for example, a cathepsin (e.g., cathepsin B).

[0233] More preferably, the linker L has the structure: TIFF2024540691000198.tif36128, where TIFF2024540691000199.tif28128 is as defined herein; * indicates attachment point to Y; and # indicates amino acid unit -W w - (if present) showing the point of attachment to the or NH group; -W w - is an amino acid unit as described herein; w is an integer as described herein, preferably w is 2, 3 or 4 (i.e., preferably -W w - is a dipeptide, tripeptide or tetrapeptide), and more preferably w is 2 or 3 (i.e., more preferably -W w - is a dipeptide or tripeptide), and in highly preferred embodiments, w is 2 (i.e., even more preferably, -W w - is a dipeptide); Q is as defined herein; m is an integer as defined herein, preferably m is 0; * denotes the point of attachment to Y; and ## denotes the point of attachment to the drug moiety (-D). Preferably, in these embodiments, the amino acid unit -W w - is a dipeptide selected from the group consisting of valine-citrulline (i.e. Val-Cit or VC), valine-alanine (i.e. Val-Ala or VA), alanine-alanine (i.e. Ala-Ala or AA) and phenylalanine-lysine (i.e. Phe-Lys or FK). More preferably, in these embodiments, the amino acid unit is a dipeptide selected from the group consisting of valine-citrulline (i.e. Val-Cit or VC), valine-alanine (i.e. Val-Ala or VA), and phenylalanine-lysine (i.e. Phe-Lys or FK). Even more preferably, in these embodiments, the amino acid unit is valine-citrulline (i.e. Val-Cit or VC) or valine-alanine (i.e. Val-Ala or VA). Even more preferably, in these embodiments, the amino acid unit is valine-citrulline (i.e. Val-Cit or VC). Alternatively, in these embodiments, the amino acid unit -W w - may be a dipeptide selected from the group consisting of valine-glutamine (i.e. Val-Gln or VQ), leucine-glutamine (i.e. Leu-Gln or LQ), phenylalanine-glutamine (i.e. Phe-Gln or FQ) and threonine-threonine (i.e. Thr-Thr or TT). In these embodiments, the amino acid unit may be a dipeptide selected from the group consisting of valine-glutamine (i.e. Val-Gln or VQ), leucine-glutamine (i.e. Leu-Gln or LQ), and phenylalanine-glutamine (i.e. Phe-Gln or FQ). In these embodiments, the amino acid unit may be valine-glutamine (i.e. Val-Gln or VQ) or leucine-glutamine (i.e. Leu-Gln or LQ). Linkers according to these embodiments may be illustrative examples for linkers that are cleavable, inter alia, by a protease, such as, for example, a cathepsin (e.g., cathepsin B).

[0234] Even more preferably, the linker L has the structure: TIFF2024540691000200.tif35128, where -W w - is an amino acid unit as described herein; w is an integer as described herein, preferably w is 2, 3 or 4 (i.e., preferably -W w - is a dipeptide, tripeptide or tetrapeptide), and more preferably w is 2 or 3 (i.e., more preferably -W w - is a dipeptide or tripeptide), and in highly preferred embodiments, w is 2 (i.e., even more preferably, -W w - is a dipeptide); * denotes the point of attachment to Y; and ## denotes the point of attachment to the drug moiety (-D). Preferably, in these embodiments, the amino acid unit -W w - is a dipeptide selected from the group consisting of valine-citrulline (i.e. Val-Cit or VC), valine-alanine (i.e. Val-Ala or VA), alanine-alanine (i.e. Ala-Ala or AA) and phenylalanine-lysine (i.e. Phe-Lys or FK). More preferably, in these embodiments, the amino acid unit is a dipeptide selected from the group consisting of valine-citrulline (i.e. Val-Cit or VC), valine-alanine (i.e. Val-Ala or VA), and phenylalanine-lysine (i.e. Phe-Lys or FK). Even more preferably, in these embodiments, the amino acid unit is valine-citrulline (i.e. Val-Cit or VC) or valine-alanine (i.e. Val-Ala or VA). Even more preferably, in these embodiments, the amino acid unit is valine-citrulline (i.e. Val-Cit or VC). Alternatively, in these embodiments, the amino acid unit -W w- may be a dipeptide selected from the group consisting of valine-glutamine (i.e. Val-Gln or VQ), leucine-glutamine (i.e. Leu-Gln or LQ), phenylalanine-glutamine (i.e. Phe-Gln or FQ) and threonine-threonine (i.e. Thr-Thr or TT). In these embodiments, the amino acid unit may be a dipeptide selected from the group consisting of valine-glutamine (i.e. Val-Gln or VQ), leucine-glutamine (i.e. Leu-Gln or LQ), and phenylalanine-glutamine (i.e. Phe-Gln or FQ). In these embodiments, the amino acid unit may be valine-glutamine (i.e. Val-Gln or VQ) or leucine-glutamine (i.e. Leu-Gln or LQ). Linkers according to these embodiments may be illustrative examples for linkers that are cleavable, inter alia, by a protease, such as, for example, a cathepsin (e.g., cathepsin B).

[0235] In a preferred embodiment, the linker L has the structure: TIFF2024540691000201.tif52129This is the amino acid unit -W w - containing the dipeptide valine-citrulline as an where * denotes the point of attachment to Y; and ## denotes the point of attachment to the drug moiety (-D). Such linkers are illustrative examples, particularly for linkers that are cleavable by a protease, such as, for example, a cathepsin (e.g., cathepsin B).

[0236] In another preferred embodiment, the linker L has the structure: TIFF2024540691000202.tif38129This is the amino acid unit -W w - containing the dipeptide valine-alanine; and where * denotes the point of attachment to Y; and ## denotes the point of attachment to the drug moiety (-D). Such linkers are illustrative examples, particularly for linkers that are cleavable by a protease, such as, for example, a cathepsin (e.g., cathepsin B).

[0237] Preferably, the linker (L) has the formula: TIFF2024540691000203.tif29128, wherein integer b is 1 and integer w is 2, 3 or 4, more preferably integer w is 2 or 3, in a highly preferred embodiment integer w is 2; and TIFF2024540691000204.tif26128 is as described herein; S is each independently a second polyalkylene glycol unit as described herein; preferably, each R S is independently a second polyethylene glycol unit as described herein; each M is independently as described herein, preferably each M is -O-; s* is an integer as described herein; preferably s* is 1; each -W-, and -B- are as defined herein; * denotes a point of attachment to Y; and ## denotes a point of attachment to the drug moiety (-D). Preferably, in these embodiments, the amino acid unit -W w - is a dipeptide selected from the group consisting of valine-citrulline (i.e. Val-Cit or VC), valine-alanine (i.e. Val-Ala or VA), alanine-alanine (i.e. Ala-Ala or AA) and phenylalanine-lysine (i.e. Phe-Lys or FK). Preferably, in these embodiments, the amino acid unit is a dipeptide selected from the group consisting of valine-citrulline (i.e. Val-Cit or VC), valine-alanine (i.e. Val-Ala or VA), and phenylalanine-lysine (i.e. Phe-Lys or FK). Even more preferably, in these embodiments, the amino acid unit is valine-citrulline (i.e. Val-Cit or VC) or valine-alanine (i.e. Val-Ala or VA). More preferably, in these embodiments, the amino acid unit is valine-citrulline (i.e. Val-Cit or VC). Alternatively, in these embodiments, the amino acid unit -W w- may be a dipeptide selected from the group consisting of valine-glutamine (i.e. Val-Gln or VQ), leucine-glutamine (i.e. Leu-Gln or LQ), phenylalanine-glutamine (i.e. Phe-Gln or FQ) and threonine-threonine (i.e. Thr-Thr or TT). In these embodiments, the amino acid unit may be a dipeptide selected from the group consisting of valine-glutamine (i.e. Val-Gln or VQ), leucine-glutamine (i.e. Leu-Gln or LQ), and phenylalanine-glutamine (i.e. Phe-Gln or FQ). In these embodiments, the amino acid unit may be valine-glutamine (i.e. Val-Gln or VQ) or leucine-glutamine (i.e. Leu-Gln or LQ). Linkers according to these embodiments may be illustrative examples for linkers that are cleavable, inter alia, by a protease, such as, for example, a cathepsin (e.g., cathepsin B).

[0238] Preferably, the linker L has the structure: TIFF2024540691000205.tif44128, where TIFF2024540691000206.tif26128 is as described herein; S is each independently a second polyalkylene glycol unit as described herein; preferably, each R S is independently a second polyethylene glycol unit as described herein; each M is independently as described herein, preferably each M is -O-; s* is an integer as described herein; preferably s* is 1; -W w - is an amino acid unit as described herein; w is an integer as described herein; preferably, w is 2, 3 or 4 (i.e., preferably, -W w - is a dipeptide, tripeptide or tetrapeptide), and more preferably w is 2 or 3 (i.e., more preferably -W w- is a dipeptide or tripeptide), and in highly preferred embodiments, w is 2 (i.e., even more preferably, -W w - is a dipeptide); Q is as described herein; m is an integer as described herein, preferably m is 0; * denotes the point of attachment to Y; and ## denotes the point of attachment to the drug moiety (-D). Preferably, in these embodiments, the amino acid unit -W w - is a dipeptide selected from the group consisting of valine-citrulline (i.e. Val-Cit or VC), valine-alanine (i.e. Val-Ala or VA), alanine-alanine (i.e. Ala-Ala or AA) and phenylalanine-lysine (i.e. Phe-Lys or FK). More preferably, in these embodiments, the amino acid unit is a dipeptide selected from the group consisting of valine-citrulline (i.e. Val-Cit or VC), valine-alanine (i.e. Val-Ala or VA), and phenylalanine-lysine (i.e. Phe-Lys or FK). Even more preferably, in these embodiments, the amino acid unit is valine-citrulline (i.e. Val-Cit or VC) or valine-alanine (i.e. Val-Ala or VA). Even more preferably, in these embodiments, the amino acid unit is valine-citrulline (i.e. Val-Cit or VC). Alternatively, in these embodiments, the amino acid unit -W w- may be a dipeptide selected from the group consisting of valine-glutamine (i.e. Val-Gln or VQ), leucine-glutamine (i.e. Leu-Gln or LQ), phenylalanine-glutamine (i.e. Phe-Gln or FQ) and threonine-threonine (i.e. Thr-Thr or TT). In these embodiments, the amino acid unit may be a dipeptide selected from the group consisting of valine-glutamine (i.e. Val-Gln or VQ), leucine-glutamine (i.e. Leu-Gln or LQ), and phenylalanine-glutamine (i.e. Phe-Gln or FQ). In these embodiments, the amino acid unit may be valine-glutamine (i.e. Val-Gln or VQ) or leucine-glutamine (i.e. Leu-Gln or LQ). Linkers according to these embodiments may be illustrative examples for linkers that are cleavable, inter alia, by a protease, such as, for example, a cathepsin (e.g., cathepsin B).

[0239] More preferably, the linker L has the structure: TIFF2024540691000207.tif35128, where TIFF2024540691000208.tif38128 is as defined herein; R S are each independently a second poly(alkylene) glycol unit as defined herein; preferably each R S is independently a second polyethylene glycol unit as defined herein; each M is independently as defined herein; preferably each M is -O-; s* is an integer as defined herein; preferably s* is 1; * indicates the point of attachment to Y; and # indicates the amino acid unit -W. w - (if present) showing the point of attachment to the or NH group; -W w - is an amino acid unit as described herein; w is an integer as described herein, preferably w is 2, 3 or 4 (i.e., preferably -W w- is a dipeptide, tripeptide or tetrapeptide), and more preferably w is 2 or 3 (i.e., more preferably -W w - is a dipeptide or tripeptide), and in highly preferred embodiments, w is 2 (i.e., even more preferably, -W w - is a dipeptide); Q is as defined herein; m is an integer as defined herein, preferably m is 0; * denotes the point of attachment to Y; and ## denotes the point of attachment to the drug moiety (-D). Preferably, in these embodiments, the amino acid unit -W w - is a dipeptide selected from the group consisting of valine-citrulline (i.e. Val-Cit or VC), valine-alanine (i.e. Val-Ala or VA), alanine-alanine (i.e. Ala-Ala or AA) and phenylalanine-lysine (i.e. Phe-Lys or FK). More preferably, in these embodiments, the amino acid unit is a dipeptide selected from the group consisting of valine-citrulline (i.e. Val-Cit or VC), valine-alanine (i.e. Val-Ala or VA), and phenylalanine-lysine (i.e. Phe-Lys or FK). Even more preferably, in these embodiments, the amino acid unit is valine-citrulline (i.e. Val-Cit or VC) or valine-alanine (i.e. Val-Ala or VA). Even more preferably, in these embodiments, the amino acid unit is valine-citrulline (i.e. Val-Cit or VC). Alternatively, in these embodiments, the amino acid unit -W w- may be a dipeptide selected from the group consisting of valine-glutamine (i.e. Val-Gln or VQ), leucine-glutamine (i.e. Leu-Gln or LQ), phenylalanine-glutamine (i.e. Phe-Gln or FQ) and threonine-threonine (i.e. Thr-Thr or TT). In these embodiments, the amino acid unit may be a dipeptide selected from the group consisting of valine-glutamine (i.e. Val-Gln or VQ), leucine-glutamine (i.e. Leu-Gln or LQ), and phenylalanine-glutamine (i.e. Phe-Gln or FQ). In these embodiments, the amino acid unit may be valine-glutamine (i.e. Val-Gln or VQ) or leucine-glutamine (i.e. Leu-Gln or LQ). Linkers according to these embodiments may be illustrative examples for linkers that are cleavable, inter alia, by a protease, such as, for example, a cathepsin (e.g., cathepsin B).

[0240] Even more preferably, the linker L has the structure: TIFF2024540691000209.tif36128, where R S are each independently a second poly(alkylene) glycol unit as defined herein; preferably, each R S is independently a second polyethylene glycol unit as defined herein; each M is independently as defined herein, preferably each M is -O-; s* is an integer as defined herein; preferably s* is 1; -W w - is an amino acid unit as described herein; w is an integer as described herein, preferably w is 2, 3 or 4 (i.e., preferably -W w - is a dipeptide, tripeptide or tetrapeptide), and more preferably w is 2 or 3 (i.e., more preferably -W w - is a dipeptide or tripeptide), and even more preferably, w is 2 (i.e., even more preferably, -Ww - is a dipeptide); * denotes the point of attachment to Y; and ## denotes the point of attachment to the drug moiety (-D). Preferably, in these embodiments, the amino acid unit -W w - is a dipeptide selected from the group consisting of valine-citrulline (i.e. Val-Cit or VC), valine-alanine (i.e. Val-Ala or VA), alanine-alanine (i.e. Ala-Ala or AA) and phenylalanine-lysine (i.e. Phe-Lys or FK). More preferably, in these embodiments, the amino acid unit is a dipeptide selected from the group consisting of valine-citrulline (i.e. Val-Cit or VC), valine-alanine (i.e. Val-Ala or VA), and phenylalanine-lysine (i.e. Phe-Lys or FK). Even more preferably, in these embodiments, the amino acid unit is valine-citrulline (i.e. Val-Cit or VC) or valine-alanine (i.e. Val-Ala or VA). Even more preferably, in these embodiments, the amino acid unit is valine-citrulline (i.e. Val-Cit or VC). Alternatively, in these embodiments, the amino acid unit -W w - may be a dipeptide selected from the group consisting of valine-glutamine (i.e. Val-Gln or VQ), leucine-glutamine (i.e. Leu-Gln or LQ), phenylalanine-glutamine (i.e. Phe-Gln or FQ) and threonine-threonine (i.e. Thr-Thr or TT). In these embodiments, the amino acid unit may be a dipeptide selected from the group consisting of valine-glutamine (i.e. Val-Gln or VQ), leucine-glutamine (i.e. Leu-Gln or LQ), and phenylalanine-glutamine (i.e. Phe-Gln or FQ). In these embodiments, the amino acid unit may be valine-glutamine (i.e. Val-Gln or VQ) or leucine-glutamine (i.e. Leu-Gln or LQ). Linkers according to these embodiments may be illustrative examples for linkers that are cleavable, inter alia, by a protease, such as, for example, a cathepsin (e.g., cathepsin B).

[0241] In a preferred embodiment, the linker L has the structure: TIFF2024540691000210.tif49128This is the amino acid unit -W w - containing the dipeptide valine-citrulline; Here, R S is a second poly(alkylene) glycol unit as defined herein; preferably, R S is a second polyethylene glycol unit as defined herein; M is as defined herein; preferably M is -O-; and * denotes the point of attachment to the Y; and ## denotes the point of attachment to the drug moiety (-D). Linkers according to these embodiments may be illustrative examples, particularly for linkers that are cleavable by a protease, such as, for example, a cathepsin (e.g., cathepsin B).

[0242] In another preferred embodiment, the linker L has the structure: TIFF2024540691000211.tif36128This is the amino acid unit -W w - containing the dipeptide valine-alanine; and Here, R S is a second poly(alkylene) glycol unit as defined herein; preferably, R S is a second polyethylene glycol unit as defined herein; M is as defined herein; preferably M is -O-; and * denotes the point of attachment to the Y; and ## denotes the point of attachment to the drug moiety (-D). Linkers according to these embodiments may be illustrative examples, particularly for linkers that are cleavable by a protease, such as, for example, a cathepsin (e.g., cathepsin B).

[0243] In some embodiments, the linker L has the formula: a -W w - ## wherein -A a- is a second spacer unit as defined herein; the integer a associated with the second spacer unit is as defined herein; -W w - is an amino acid unit as defined herein; the integer w associated with the amino acid unit W is as defined herein; the first spacer unit (-B b -) is absent; * denotes a point of attachment to Y; and # denotes a point of attachment to the drug moiety (-D). Preferably, the integer a is 1. Preferably, the integer w is 2, 3 or 4, more preferably the integer w is 2 or 3, and even more preferably the integer w is 2. Preferably, in these embodiments, the amino acid unit -W w - is a dipeptide selected from the group consisting of valine-citrulline (i.e. Val-Cit or VC), valine-alanine (i.e. Val-Ala or VA), alanine-alanine (i.e. Ala-Ala or AA) and phenylalanine-lysine (i.e. Phe-Lys or FK). More preferably, in these embodiments, the amino acid unit is a dipeptide selected from the group consisting of valine-citrulline (i.e. Val-Cit or VC), valine-alanine (i.e. Val-Ala or VA), and phenylalanine-lysine (i.e. Phe-Lys or FK). Even more preferably, in these embodiments, the amino acid unit is valine-citrulline (i.e. Val-Cit or VC) or valine-alanine (i.e. Val-Ala or VA). Even more preferably, in these embodiments, the amino acid unit is valine-citrulline (i.e. Val-Cit or VC). Alternatively, in these embodiments, the amino acid unit -W w- may be a dipeptide selected from the group consisting of valine-glutamine (i.e., Val-Gln or VQ), leucine-glutamine (i.e., Leu-Gln or LQ), phenylalanine-glutamine (i.e., Phe-Gln or FQ) and threonine-threonine (i.e., Thr-Thr or TT). In these embodiments, the amino acid unit may be a dipeptide selected from the group consisting of valine-glutamine (i.e., Val-Gln or VQ), leucine-glutamine (i.e., Leu-Gln or LQ), and phenylalanine-glutamine (i.e., Phe-Gln or FQ). In these embodiments, the amino acid unit may be valine-glutamine (i.e., Val-Gln or VQ) or leucine-glutamine (i.e., Leu-Gln or LQ). In any one of these embodiments, the second spacer unit -A- has the structure: TIFF2024540691000212.tif26128, where TIFF2024540691000213.tif26128 is as defined herein.

[0244] The linker L may have the structure: TIFF2024540691000214.tif27128, where TIFF2024540691000215.tif26128 is as defined herein; * indicates attachment point to Y; and # indicates amino acid unit -W w - Shows connection points; -W w - is an amino acid unit as described herein; w is an integer as described herein, preferably w is 2, 3 or 4 (i.e., preferably -W w - is a dipeptide, tripeptide or tetrapeptide), and more preferably, the integer w is 2 or 3 (i.e., more preferably, -W w - is a dipeptide or tripeptide), and even more preferably, w is 2 (i.e., even more preferably, -W w - is a dipeptide); * denotes the point of attachment to Y; and ## denotes the point of attachment to the drug moiety (-D). Preferably, in these embodiments, the amino acid unit -W w - is a dipeptide selected from the group consisting of valine-citrulline (i.e. Val-Cit or VC), valine-alanine (i.e. Val-Ala or VA), alanine-alanine (i.e. Ala-Ala or AA) and phenylalanine-lysine (i.e. Phe-Lys or FK). More preferably, in these embodiments, the amino acid unit is a dipeptide selected from the group consisting of valine-citrulline (i.e. Val-Cit or VC), valine-alanine (i.e. Val-Ala or VA), and phenylalanine-lysine (i.e. Phe-Lys or FK). Even more preferably, in these embodiments, the amino acid unit is valine-citrulline (i.e. Val-Cit or VC) or valine-alanine (i.e. Val-Ala or VA). Even more preferably, in these embodiments, the amino acid unit is valine-citrulline (i.e. Val-Cit or VC). Alternatively, in these embodiments, the amino acid unit -W w - can be a dipeptide selected from the group consisting of valine-glutamine (i.e., Val-Gln or VQ), leucine-glutamine (i.e., Leu-Gln or LQ), phenylalanine-glutamine (i.e., Phe-Gln or FQ), and threonine-threonine (i.e., Thr-Thr or TT). In these embodiments, the amino acid unit can be a dipeptide selected from the group consisting of valine-glutamine (i.e., Val-Gln or VQ), leucine-glutamine (i.e., Leu-Gln or LQ), and phenylalanine-glutamine (i.e., Phe-Gln or FQ). In these embodiments, the amino acid unit can be valine-glutamine (i.e., Val-Gln or VQ) or leucine-glutamine (i.e., Leu-Gln or LQ).

[0245] In some embodiments, the linker L can have the structure: TIFF2024540691000216.tif39128This is the amino acid unit -W w- containing the dipeptide valine-citrulline as an where * denotes the point of attachment to the Y; and ## denotes the point of attachment to the drug moiety (-D).

[0246] In some embodiments, the linker L can have the structure: TIFF2024540691000217.tif28128This is the amino acid unit -W w - containing the dipeptide valine-alanine; and where * denotes the point of attachment to the Y; and ## denotes the point of attachment to the drug moiety (-D).

[0247] In some embodiments, the linker (-L-) has the formula: a - ## wherein -A- is a second spacer unit as defined herein; the integer a associated with the second spacer unit is 1; w - is absent; the first spacer unit (-B-) is absent; * denotes a point of attachment to Y; and ## denotes a point of attachment to the drug moiety (-D). In any one of these embodiments, the second spacer unit -A a -The structure: TIFF2024540691000218.tif26128, where TIFF2024540691000219.tif26128 is as defined herein.

[0248] The linker (-L-) can have the structure: TIFF2024540691000220.tif22128, where TIFF2024540691000221.tif22128 is as defined herein; * indicates the point of attachment to the Y; and # indicates the point of attachment to the drug moiety (-D).

[0249] In some embodiments, the linker L can have the structure: TIFF2024540691000222.tif25128 where * indicates the point of attachment to Y; and ## indicates the point of attachment to the drug moiety (-D).

[0250] Linker*-A a -Q CO q -G- ## In some embodiments, the linker L has the following structure: a -Q CO q -G- ## where -A- is a second spacer unit, as described herein; a is 0 or 1, as described herein; and each -Q CO - is independently a connector unit; q is 0 or 1; and -G- is a first spacer unit comprising a sugar moiety; * denotes a point of attachment to Y; and ## denotes a point of attachment to a drug moiety (-D). Linkers comprising a sugar moiety, e.g., a glucuronic acid moiety, are described, e.g., in Jeffrey et al., "Development and Properties of beta-Glucuronide Linkers for Monoclonal Antibody-Drug Conjugates", Bioconjugate Chem. 2006, 17, 831-840, doi: 10.1021 / bc0600214; WO 2019 / 236954; and WO 2015 / 057699.

[0251] Linker*-A a -Q CO q -G- ## In the structure *-A, the second spacer unit -A-, when present, can be any second spacer unit as described herein. a -Q CO q -G- ## In a linker having the formula: CO(if present) and / or the first spacer unit containing a sugar moiety. The second spacer unit (-A-), if present, serves to connect Y to a connector unit (Q CO ) can be any chemical group or moiety that can be linked to a connector unit Q CO In the absence of a spacer unit, Y may be linked to a first spacer unit (-G-) that includes a sugar moiety. In this regard, Y is attached to a second spacer unit (-A-) as described herein. The second spacer unit (-A-) may be linked to a connector unit (-Q CO Depending on whether the connector unit (-Q CO -) or to a first spacer unit (-G-) bearing a sugar moiety. Preferably, the connector unit (-Q CO A functional group capable of forming a bond to the first spacer unit (-G-) or to the first spacer unit (-G-) that comprises a sugar moiety can be, for example: TIFF2024540691000223.tif14128 or a carbonyl group depicted as -C(O)-. The integer a associated with the second spacer unit can be 0 or 1. Preferably, the integer a is 1. Alternatively, in other embodiments, the second spacer unit is absent (a = 0).

[0252] Linker*-A a -Q CO q -G- ## In the formula (I), the second spacer unit -A-, when present, can be any second spacer unit as described herein. In some embodiments, the second spacer unit -A-, when present, has the structure: TIFF2024540691000224.tif26128, where TIFF2024540691000225.tif26128 is as defined herein. Thus, in some embodiments, the linker (L) has the structure: TIFF2024540691000226.tif29128, where L P , R S ,s*,M,Q CO , q, and G are as defined herein; * denotes the point of attachment to -Y-; and ## denotes the point of attachment to the drug moiety (-D).

[0253] Connector unit (-Q CO -) may be included when it is desirable to add additional distance between -Y- or, if present, the second spacer unit (-A-) and the first spacer unit (-G-) that includes the sugar moiety. In some embodiments, the extra distance may aid in activation within the first spacer unit (-G-) that includes the sugar moiety. Thus, the connector unit (-Q CO -), if present, extends the framework of the linker (-L-). In this regard, the connector unit (-Q CO -) is covalently linked at one end to -Y- or, if a second spacer unit -A- is present, to a second spacer unit (-A-), and is linked at the other end to a connector unit (-Q CO -) is covalently attached at its other end to a first spacer unit (-G-) that contains a sugar moiety. CO The integer q associated with can be 0 or 1. Preferably, the integer q is 1. Alternatively, in other embodiments, the connector unit Q CO does not exist (q = 0).

[0254] Connector unit (-Q CO The connector unit Q serves to link the first spacer unit (-G-), which contains the sugar moiety, to the second spacer unit (-A-), if present, or to -Y-. COcan be any chemical group or moiety that serves to provide a connection of the first spacer unit (-G-), including the sugar moiety, to the second spacer unit (-A-), if present, or to -Y-. The connector unit can be composed of, for example, one or more (e.g., 1-10, preferably 1, 2, 3, or 4) natural or unnatural amino acids, amino alcohols, amino aldehydes, and diamino residues. In some embodiments, the connector unit (-Q CO -) is a single natural or unnatural amino acid, amino alcohol, amino aldehyde, or diamino residue. In some embodiments, the amino acid that can act as a connector unit is β-alanine. In particular, the connector unit can be a single β-alanine.

[0255] In some embodiments, the connector unit (-Q CO -) has the formula shown below: TIFF2024540691000227.tif98135 where the wavy line indicates the connection of a connector unit within the linker (-L-) or, if the second spacer unit (-A-) is absent, to -Y-; and where R 111 is hydrogen, hydroxybenzyl, methyl, isopropyl, isobutyl, sec-butyl, -CH 2 OH, -CH(OH)CH 3 , -CH 2 CH 2 SCH 3 , -CH 2 CONH 2 , -CH 2 COOH, -CH 2 CH 2 CONH 2 , -CH 2 CH 2 COOH, -(CH 2 ) 3 NHC(=NH)NH 2 , -(CH 2 ) 3 NH 2 , -(CH 2 ) 3 NHCOCH 3 , -(CH2 ) 3 NHCHO, -(CH 2 ) 4 NHC(=NH)NH 2 , -(CH 2 ) 4 NH 2 , -(CH 2 ) 4 NHCOCH 3 , -(CH 2 ) 4 NHCHO, -(CH 2 ) 3 NHCONH 2 , -(CH 2 ) 4 NHCONH 2 , -CH 2 CH 2 CH(OH)CH 2 NH 2 , 2-pyridylmethyl-, 3-pyridylmethyl-, 4-pyridylmethyl-, TIFF2024540691000228.tif25149, and each R 100 is hydrogen or -(C 1 -C 3 ) alkyl, preferably hydrogen or CH 3 and subscript c is an independently selected integer from 1 to 10, preferably 1 to 3.

[0256] In a preferred embodiment, the connector unit has the following structure: CO -), which has a carbonyl group for connection to a first spacer unit (-G-) that comprises a sugar moiety, and an NH group for connection to a second spacer unit (-A-), if present, as follows: TIFF2024540691000229.tif23128 where, in each case, R 13 is -(C 1 -C 6 ) alkylene-, -(C 3 -C 8 )Carbocyclo-, -arylene-, -(C 1 -C 10 )Heteroalkylene-, -(C 3-C 8 )Heterocyclo-, -(C 1 -C 10 ) alkylene-arylene-, -arylene-(C 1 -C 10 ) alkylene-, -(C 1 -C 10 )Alkylene-(C 3 -C 8 )Carbocyclo)-, -(C 3 -C 8 )Carbocyclo-(C 1 -C 10 ) alkylene-, -(C 1 -C 10 )Alkylene-(C 3 -C 8 )heterocyclo-, and -(C 3 -C 8 )Heterocyclo-(C 1 -C 10 ) alkylene-, and the subscript c is an integer ranging from 1 to 4. In some embodiments, R 13 Ha-(C 1 -C 6 ) alkylene, and c is an integer ranging from 1 to 4. In a preferred embodiment, R 13 Ha-(C 1 -C 6 ) alkylene and c is 1.

[0257] More preferably, the connector unit (-Q CO -) has the following structure: TIFF2024540691000230.tif17128 where the wavy line adjacent to the nitrogen indicates a covalent connection to the second spacer unit (-A-) (if present) and the wavy line adjacent to the carbonyl indicates a covalent connection to the first spacer group (-G-) that comprises a sugar moiety; and m is an integer in the range of 1 to 6, preferably 2 to 6, more preferably 2 to 4.

[0258] Even more preferably, the connector unit (-Q CO -) has the following structure: TIFF2024540691000231.tif13128 where the wavy line adjacent to the nitrogen indicates the covalent connection to the second spacer unit (-A-), if present, and the wavy line adjacent to the carbonyl indicates the covalent connection to the first spacer group (-G-), which comprises a sugar moiety.

[0259] Another representative connector unit (-Q) having a carbonyl group for connection to a first spacer unit (-G-) containing a sugar moiety. CO -) is as follows: TIFF2024540691000232.tif12128, where R 13 is -(C 1 -C 6 ) alkylene-, -(C 3 -C 8 )Carbocyclo-, -arylene-, -(C 1 -C 10 )Heteroalkylene-, -(C 3 -C 8 )Heterocyclo-, -(C 1 -C 10 ) alkylene-arylene-, -arylene-(C 1 -C 10 ) alkylene-, -(C 1 -C 10 )Alkylene-(C 3 -C 8 )Carbocyclo-, -(C 3 -C 8 )Carbocyclo-(C 1 -C 10 ) alkylene-, -(C 1 -C 10 )Alkylene-(C 3 -C 8 )heterocyclo-, or -(C 3 -C 8 )Heterocyclo-(C 1 -C 10 ) alkylene-. In some embodiments, R 13 Ha-(C 1 -C 6 ) alkylene.

[0260] Another representative connector unit having an NH moiety connecting to a first spacer unit (-G-) that includes a sugar moiety is: TIFF2024540691000233.tif23128 where, in each case, R 13 is -(C1-C6) alkylene-, -(C 3 -C 8 )Carbocyclo-, -arylene-, -(C 1 -C 10 )Heteroalkylene-, -(C 3 -C 8 )Heterocyclo-, -(C 1 -C 10 ) alkylene-arylene-, -arylene-(C 1 -C 10 ) alkylene-, -(C 1 -C 10 )Alkylene-(C 3 -C 8 )Carbocyclo-, -(C 3 -C 8 )Carbocyclo-(C 1 -C 10 ) alkylene-, -(C 1 -C 10 )Alkylene-(C 3 -C 8 )heterocyclo-, and -(C 3 -C 8 )Heterocyclo-(C 1 -C 10 ) alkylene-, and subscript c is 1 to 14. In some embodiments, R 13 Ha-(C 1 -C 6 ) alkylene and subscript c is 1.

[0261] Another representative connector unit (-Q) having an NH moiety connected to a first spacer unit (-G-) containing a sugar moiety. CO -) is as follows: TIFF2024540691000234.tif7128, where R 13 is -(C 1 -C 6) alkylene-, -(C 3 -C 8 )Carbocyclo-, -arylene-, -(C 1 -C 10 )Heteroalkylene-, -(C 3 -C 8 )Heterocyclo-, -(C 1 -C 10 ) alkylene-arylene-, -arylene-(C 1 -C 10 ) alkylene-, -(C 1 -C 10 )Alkylene-(C 3 -C 8 )Carbocyclo-, -(C 3 -C 8 )Carbocyclo-(C 1 -C 10 ) alkylene-, -(C 1 -C 10 )Alkylene-(C 3 -C 8 )Heterocyclo-, -(C 3 -C 8 )Heterocyclo-(C 1 -C 10 ) alkylene-, -C(=O)(C 1 -C 10 ) alkylene- or -(C 1 -C 6 ) alkylene-C(=O)-(C 1 -C 6 ) alkylene.

[0262] A first spacer unit (-G-) having a sugar moiety must be present, structure *-A a -Q CO q -G- ## In some embodiments, the first spacer unit (-G-) comprising a sugar moiety forms a cleavable bond with the drug moiety (-D). In some embodiments, the first spacer unit (-G-) comprising a sugar moiety is the only component of the linker having a connector unit (-Q CO-) (if present). In some embodiments, the cleavable bond is within the first spacer unit (-G-) that includes a sugar moiety, but allows for release of the free drug (e.g., by a 1,6-elimination reaction after cleavage). Functional groups for forming cleavable bonds can include, for example, sugars for forming glycosidic bonds.

[0263] The structure and sequence of the first spacer unit (-G-) containing a sugar moiety can be such that the unit is cleaved by the action of an enzyme present at the target site. In other embodiments, the first spacer unit (-G-) containing a sugar moiety can be cleavable by other mechanisms. The first spacer unit (-G-) containing a sugar moiety can include one or more cleavage sites.

[0264] Preferably, the first spacer unit comprising a sugar moiety (-G-) comprises a sugar cleavage site. In some such embodiments, the first spacer unit comprising a sugar moiety (-G-) comprises a sugar moiety (Su) linked to a self-immolative group via an oxygen glycosidic bond. In such aspects, the self-immolative group is considered to be part of the first spacer unit comprising a sugar moiety (-G-). In this regard, a "self-immolative group" refers to a group consisting of three spaced chemical moieties, i.e., a sugar moiety (via a glycosidic bond), a drug moiety (-D), and a -Q CO Depending on whether -unit and / or -A-unit are present, connector unit -Q CO The glycosidic bond may be a trifunctional chemical moiety capable of covalently linking together the first spacer unit (-A-, the second spacer unit -A-, or -Y-). The glycosidic bond may be one that can be cleaved at the target site to initiate a self-immolative reaction sequence leading to the release of the drug. The particular sugar moiety may be selected, for example, from the group consisting of glucuronic acid, galactose, glucose, arabinose, mannose-6-phosphate, fucose, rhamnose, gulose, allose, 6-deoxy-glucose, lactose, maltose, cellobiose, gentiobiose, maltotriose, GlcNAc, GalNAc, and maltohexaose.

[0265] Thus, the first spacer unit (-G-) comprising a sugar moiety has the formula: TIFF2024540691000235.tif21128, where the self-immolative group K forms a covalent bond with the drug moiety, and -Q CO Forms a covalent bond with -, -A-, or -Y- (as appropriate).

[0266] The first spacer unit (-G-) comprising a sugar moiety can, for example, be represented by the formula: TIFF2024540691000236.tif47128, where Su is a sugar moiety, -O'- represents an oxygen glycosidic bond; each R is independently hydrogen, halogen, -CN, or -NO 2 and the wavy line is -Q CO -, -A-, or -Y- (as appropriate), and the asterisk indicates the connection to the drug moiety (directly or indirectly through a spacer unit; the spacer unit, if present, can be, for example, -(C=O)-).

[0267] In some such embodiments, the sugar cleavage site is recognized by β-glucuronidase and the first spacer unit (-G-) that comprises a sugar moiety comprises a glucuronide unit. The glucuronide unit has the formula: TIFF2024540691000237.tif20128, wherein the glucuronic acid is linked to a self-immolative group (K) via a glycosidic bond (-O'-); wherein the self-immolative group K is covalently attached to the drug moiety (either directly or indirectly through a spacer unit; the spacer unit, if present, can be, for example, -(C=O)-), and -Q CO Forms a covalent bond with -, -A-, or -Y- (as appropriate).

[0268] The glucuronide unit can be, for example, of the formula: TIFF2024540691000238.tif36128, In the formula, the wavy line represents -Q CO -, -A-, or -Y- (as appropriate), and an asterisk indicates a covalent attachment to a drug moiety, -C (directly or indirectly via a spacer unit; the spacer unit, if present, can be, for example, -(C=O)-).

[0269] In some embodiments, the first spacer unit comprising a sugar moiety (-G-) comprises a sugar cleavage site, -SC, i.e., the combination of the first spacer unit comprising a sugar moiety (-G-) and the drug moiety (-D) is represented by the formula: TIFF2024540691000239.tif47128TIFF2024540691000240.tif41128In the formula, Su is a sugar moiety, D is a drug moiety, -O'- represents an oxygen glycosidic bond; each R is independently hydrogen or halogen, -CN, -NO 2 or other electron-withdrawing group, -Q CO - is a connector unit as described herein; where the wavy bond indicates a covalent connection to -A- or -Y- (as appropriate).

[0270] When the first spacer unit comprising a sugar moiety (-G-) comprises a glucuronide unit, -SC, i.e., the combination of the first spacer unit comprising a sugar moiety (-G-) and the drug moiety (-D), can be represented, for example, by the following formula: TIFF2024540691000241.tif76128 where the wavy bond indicates a covalent connection to -A- or -Y- (as appropriate); D is a drug moiety; and -Q CO - is a connector unit as described herein.

[0271] In a preferred embodiment, the linker (L) has the following structure: TIFF2024540691000242.tif59128, where -A- is a second spacer unit as described herein; a is an integer as described herein, preferably a is 1; -Q CO - is a connector unit as described herein; q is an integer as defined herein, preferably q is 1; * denotes a point of attachment to the Y; and ## denotes a point of attachment to the drug moiety (-D). In these embodiments, the connector unit (Q CO ), if present, may have a carbonyl group for connection to the first spacer unit (-G-), which comprises a sugar moiety, and an NH group for connection to the second spacer unit (-A-), if present, may be as follows: TIFF2024540691000243.tif23128 where, in each case, R 13 is -(C 1 -C 6 ) alkylene-, -(C 3 -C 8 )Carbocyclo-, -arylene-, -(C 1 -C 10 )Heteroalkylene-, -(C 3 -C 8 )Heterocyclo-, -(C 1 -C 10 ) alkylene-arylene-, -arylene-(C 1 -C 10 ) alkylene-, -(C 1 -C 10 )Alkylene-(C 3 -C 8 )Carbocyclo)-, -(C 3 -C 8 )Carbocyclo-(C 1 -C 10 ) alkylene-, -(C 1 -C 10 )Alkylene-(C 3 -C 8 )heterocyclo-, and -(C 3 -C 8 )Heterocyclo-(C 1 -C 10) alkylene-, and the subscript c is an integer ranging from 1 to 4. In some embodiments, R 13 Ha-(C 1 -C 6 ) alkylene, and c is an integer ranging from 1 to 4. In a preferred embodiment, R 13 Ha-(C 1 -C 6 ) alkylene and c is 1; preferably, in these embodiments, the connector unit (-Q CO -), when present, can have the structure: TIFF2024540691000244.tif16128 Here, the wavy line adjacent to the nitrogen represents the second spacer unit (-A a -), when present, and the wavy line adjacent to the carbonyl indicates a covalent connection to a first spacer group (-G-) that comprises a sugar moiety; and m is an integer ranging from 1 to 6, preferably 2 to 6, more preferably 2 to 4; more preferably, in these embodiments, a connector unit (-Q CO -), when present, has the structure: TIFF2024540691000245.tif13128 Here, the wavy line adjacent to the nitrogen represents the second spacer unit (-A a The wavy line indicates the covalent attachment to the carbonyl (-G-), if present, and adjacent to the carbonyl indicates the covalent attachment to the first spacer group (-G-), which comprises a sugar moiety.

[0272] More preferably, the linker (L) has the following structure: TIFF2024540691000246.tif58128, where TIFF2024540691000247.tif22128 is as defined herein; * indicates a connection point to -Y-; and # indicates a connector unit (-Q CO -) (if present) or to indicate the point of attachment to the NH group; -Q CO- is a connector unit as defined herein; q is an integer as defined herein, preferably q is 1; * denotes a point of attachment to the Y; and ## denotes a point of attachment to the drug moiety (-D). In these embodiments, the connector unit (Q CO ), if present, can have a carbonyl group for connection to the first spacer unit (-G-) that includes a sugar moiety and an NH group for connection to the second spacer unit (-A-), and can be as follows: TIFF2024540691000248.tif23128 where, in each case, R 13 is -(C 1 -C 6 ) alkylene-, -(C 3 -C 8 )Carbocyclo-, -arylene-, -(C 1 -C 10 )Heteroalkylene-, -(C 3 -C 8 )Heterocyclo-, -(C 1 -C 10 ) alkylene-arylene-, -arylene-(C 1 -C 10 ) alkylene-, -(C 1 -C 10 )Alkylene-(C 3 -C 8 )Carbocyclo)-, -(C 3 -C 8 )Carbocyclo-(C 1 -C 10 ) alkylene-, -(C 1 -C 10 )Alkylene-(C 3 -C 8 )heterocyclo-, and -(C 3 -C 8 )Heterocyclo-(C 1 -C 10 ) alkylene-, and the subscript c is an integer ranging from 1 to 4. In some embodiments, R 13 Ha-(C 1 -C 6) alkylene, and c is an integer ranging from 1 to 4. In a preferred embodiment, R 13 Ha-(C 1 -C 6 ) alkylene and c is 1. Preferably, in these embodiments, the connector unit (-Q CO p -), when present, can have the structure: TIFF2024540691000249.tif17128, wherein the wavy line adjacent to the nitrogen indicates a covalent connection to a second spacer unit (-A-), and the wavy line adjacent to the carbonyl indicates a covalent connection to a first spacer group (-G-) that includes a sugar moiety; and m is an integer ranging from 1 to 6, preferably 2 to 6, more preferably 2 to 4. More preferably, in these embodiments, the connector unit (-Q CO -), when present, has the structure: TIFF2024540691000250.tif13128 Here, the wavy line adjacent to the nitrogen represents the second spacer unit (-A a The wavy line indicates the covalent attachment to the carbonyl (-G-), if present, and adjacent to the carbonyl indicates the covalent attachment to the first spacer group (-G-), which comprises a sugar moiety.

[0273] Even more preferably, the linker (L) has the structure: TIFF2024540691000251.tif59128, where Q CO is a connector unit as defined herein; q is an integer as defined herein, preferably q is 1; * denotes a point of attachment to the Y; and ## denotes a point of attachment to the drug moiety (-D). In these embodiments, the connector unit (Q CO ), if present, can have a carbonyl group for connection to the first spacer unit (-G-) that includes a sugar moiety and an NH group for connection to the second spacer unit (-A-), and can be as follows: TIFF2024540691000252.tif23128 where, in each case, R 13is -(C 1 -C 6 ) alkylene-, -(C 3 -C 8 )Carbocyclo-, -arylene-, -(C 1 -C 10 )Heteroalkylene-, -(C 3 -C 8 )Heterocyclo-, -(C 1 -C 10 ) alkylene-arylene-, -arylene-(C 1 -C 10 ) alkylene-, -(C 1 -C 10 )Alkylene-(C 3 -C 8 )Carbocyclo)-, -(C 3 -C 8 )Carbocyclo-(C 1 -C 10 ) alkylene-, -(C 1 -C 10 )Alkylene-(C 3 -C 8 )heterocyclo-, and -(C 3 -C 8 )Heterocyclo-(C 1 -C 10 ) alkylene-, and the subscript c is an integer ranging from 1 to 4. In some embodiments, R 13 Ha-(C 1 -C 6 ) alkylene, and c is an integer ranging from 1 to 4. In a preferred embodiment, R 13 Ha-(C 1 -C 6 ) alkylene and c is 1. Preferably, in these embodiments, the connector unit (-Q CO -), when present, can have the structure: TIFF2024540691000253.tif16128, wherein the wavy line adjacent to the nitrogen indicates a covalent connection to a second spacer unit (-A-), and the wavy line adjacent to the carbonyl indicates a covalent connection to a first spacer group (-G-) that includes a sugar moiety; and m is an integer ranging from 1 to 6, preferably 2 to 6, more preferably 2 to 4; more preferably, in these embodiments, a connector unit (-Q CO -), when present, has the structure: TIFF2024540691000254.tif13128 where the wavy line adjacent to the nitrogen indicates the covalent connection to the second spacer unit (-A-) and the wavy line adjacent to the carbonyl indicates the covalent connection to the first spacer group (-G-), which comprises a sugar moiety.

[0274] Even more preferably, the linker L may have the structure: TIFF2024540691000255.tif51128 where * denotes the point of attachment to -Y-; and ## denotes the point of attachment to the drug moiety (-D).

[0275] In a preferred embodiment, the linker L has the structure: TIFF2024540691000256.tif60128, where TIFF2024540691000257.tif26128 is as described herein; S is each independently a second polyalkylene glycol unit as described herein; preferably, each R S is independently a second polyethylene glycol unit as described herein; each M is independently as described herein; preferably each M is -O-; s* is an integer as described herein; preferably s* is 1; -Q CO - is a connector unit as described herein; q is an integer as defined herein, preferably q is 1; * denotes a point of attachment to the Y; and ## denotes a point of attachment to the drug moiety (-D). In these embodiments, the connector unit (Q CO ), if present, can have a carbonyl group for connection to the first spacer unit (-G-) that includes a sugar moiety and an NH group for connection to the second spacer unit (-A-), and can be as follows: TIFF2024540691000258.tif23128 where, in each case, R 13 is -(C 1 -C 6 ) alkylene-, -(C 3 -C 8 )Carbocyclo-, -arylene-, -(C 1 -C 10 )Heteroalkylene-, -(C 3 -C 8 )Heterocyclo-, -(C 1 -C 10 ) alkylene-arylene-, -arylene-(C 1 -C 10 ) alkylene-, -(C 1 -C 10 )Alkylene-(C 3 -C 8 )Carbocyclo)-, -(C 3 -C 8 )Carbocyclo-(C 1 -C 10 ) alkylene-, -(C 1 -C 10 )Alkylene-(C 3 -C 8 )heterocyclo-, and -(C 3 -C 8 )Heterocyclo-(C 1 -C 10 ) alkylene-, and the subscript c is an integer ranging from 1 to 4. In some embodiments, R 13 Ha-(C 1 -C 6 ) alkylene, and c is an integer ranging from 1 to 4. In a preferred embodiment, R 13 Ha-(C 1 -C 6) alkylene and c is 1. Preferably, in these embodiments, the connector unit (-Q CO -), when present, can have the structure: TIFF2024540691000259.tif16128, wherein the wavy line adjacent to the nitrogen indicates a covalent connection to a second spacer unit (-A-), and the wavy line adjacent to the carbonyl indicates a covalent connection to a first spacer group (-G-) that includes a sugar moiety; and m is an integer ranging from 1 to 6, preferably 2 to 6, more preferably 2 to 4. More preferably, in these embodiments, the connector unit (-Q CO -), when present, has the structure: TIFF2024540691000260.tif13128 where the wavy line adjacent to the nitrogen indicates the covalent connection to the second spacer unit (-A-) and the wavy line adjacent to the carbonyl indicates the covalent connection to the first spacer group (-G-), which comprises a sugar moiety.

[0276] More preferably, the linker (L) has the following structure: TIFF2024540691000261.tif57128, where TIFF2024540691000262.tif30128 is as defined herein; R S are each independently a second poly(alkylene) glycol unit as defined herein; preferably, each R S is independently a second polyethylene glycol unit as defined herein; each M is independently as defined herein; preferably each M is -O-; s* is an integer as defined herein; preferably s* is 1; * denotes a point of attachment to Y; * denotes a point of attachment to -Y-; and # denotes a connector unit (-Q CO -) (if present) or to indicate the point of attachment to the NH group; -Q CO - is a connector unit as defined herein; q is an integer as defined herein, preferably q is 1; * denotes a point of attachment to the Y; and ## denotes a point of attachment to the drug moiety (-D). In these embodiments, the connector unit (Q CO ), if present, can have a carbonyl group for connection to the first spacer unit (-G-) that includes a sugar moiety and an NH group for connection to the second spacer unit (-A-), and can be as follows: TIFF2024540691000263.tif23128 where, in each case, R 13 is -(C 1 -C 6 ) alkylene-, -(C 3 -C 8 )Carbocyclo-, -arylene-, -(C 1 -C 10 )Heteroalkylene-, -(C 3 -C 8 )Heterocyclo-, -(C 1 -C 10 ) alkylene-arylene-, -arylene-(C 1 -C 10 ) alkylene-, -(C 1 -C 10 )Alkylene-(C 3 -C 8 )Carbocyclo)-, -(C 3 -C 8 )Carbocyclo-(C 1 -C 10 ) alkylene-, -(C 1 -C 10 )Alkylene-(C 3 -C 8 )heterocyclo-, and -(C 3 -C 8 )Heterocyclo-(C 1 -C 10 ) alkylene-, and the subscript c is an integer ranging from 1 to 4. In some embodiments, R 13 Ha-(C 1 -C 6 ) alkylene, and c is an integer ranging from 1 to 4. In a preferred embodiment, R 13 Ha-(C 1 -C 6) alkylene and c is 1. Preferably, in these embodiments, the connector unit (-Q CO -) may have the structure: TIFF2024540691000264.tif17128, wherein the wavy line adjacent to the nitrogen indicates a covalent connection to a second spacer unit (-A-), and the wavy line adjacent to the carbonyl indicates a covalent connection to a first spacer group (-G-) that includes a sugar moiety; and m is an integer ranging from 1 to 6, preferably 2 to 6, more preferably 2 to 4. More preferably, in these embodiments, the connector unit (-Q CO -), when present, has the structure: TIFF2024540691000265.tif13128 where the wavy line adjacent to the nitrogen indicates the covalent connection to the second spacer unit (-A-) and the wavy line adjacent to the carbonyl indicates the covalent connection to the first spacer group (-G-), which comprises a sugar moiety.

[0277] More preferably, the linker (L) has the following structure: TIFF2024540691000266.tif60128, where R S is each independently a second polyalkylene glycol unit as defined herein; preferably, each R S is independently a second polyethylene glycol unit as described herein; each M is independently as defined herein; preferably each M is -O-; s* is an integer as defined herein; preferably s* is 1. Q CO is a connector unit as defined herein; q is an integer as defined herein, preferably q is 1; * denotes a point of attachment to the Y; and ## denotes a point of attachment to the drug moiety (-D). In these embodiments, the connector unit (Q CO), if present, can have a carbonyl group for connection to the first spacer unit (-G-) that includes a sugar moiety and an NH group for connection to the second spacer unit (-A-), and can be as follows: TIFF2024540691000267.tif23128 where, in each case, R 13 is -(C 1 -C 6 ) alkylene-, -(C 3 -C 8 )Carbocyclo-, -arylene-, -(C 1 -C 10 )Heteroalkylene-, -(C 3 -C 8 )Heterocyclo-, -(C 1 -C 10 ) alkylene-arylene-, -arylene-(C 1 -C 10 ) alkylene-, -(C 1 -C 10 )Alkylene-(C 3 -C 8 )Carbocyclo)-, -(C 3 -C 8 )Carbocyclo-(C 1 -C 10 ) alkylene-, -(C 1 -C 10 )Alkylene-(C 3 -C 8 )heterocyclo-, and -(C 3 -C 8 )Heterocyclo-(C 1 -C 10 ) alkylene-, and the subscript c is an integer ranging from 1 to 4. In some embodiments, R 13 Ha-(C 1 -C 6 ) alkylene, and c is an integer ranging from 1 to 4. In a preferred embodiment, R 13 Ha-(C 1 -C 6 ) alkylene and c is 1. Preferably, in these embodiments, the connector unit (-Q CO -), when present, can have the structure: TIFF2024540691000268.tif16128, wherein the wavy line adjacent to the nitrogen indicates a covalent connection to a second spacer unit (-A-), and the wavy line adjacent to the carbonyl indicates a covalent connection to a first spacer group (-G-) that includes a sugar moiety; and m is an integer ranging from 1 to 6, preferably 2 to 6, more preferably 2 to 4. More preferably, in these embodiments, the connector unit (-Q CO -), when present, has the structure: TIFF2024540691000269.tif13128 where the wavy line adjacent to the nitrogen indicates the covalent connection to the second spacer unit (-A-) and the wavy line adjacent to the carbonyl indicates the covalent connection to the first spacer group (-G-), which comprises a sugar moiety.

[0278] Even more preferably, the linker L has the structure: TIFF2024540691000270.tif50128, where R S is each independently a second polyalkylene glycol unit as defined herein; preferably, each R S is independently a second polyethylene glycol unit as described herein; each M is independently as defined herein; preferably each M is -O-; s* is an integer as defined herein; preferably s* is 1. * denotes the point of attachment to -Y-; and ## denotes the point of attachment to the drug moiety (-D).

[0279] Also preferably, the linker L has the structure: TIFF2024540691000271.tif58128 where * denotes the point of attachment to -Y-; and ## denotes the point of attachment to the drug moiety (-D).

[0280] In one embodiment, the linker L has the structure: TIFF2024540691000272.tif58128 where * denotes the point of attachment to -Y-; and ## denotes the point of attachment to the drug moiety (-D). In this embodiment, Y may be as defined herein; preferably, Y may be NH.

[0281] Linker*-A a -U AT u -Sulf- ## In some embodiments, the linker L has the structure: a -U AT u -Sulf- ## wherein: -A- is a second spacer unit; a is 0 or 1; each -U AT u - is independently a connecting unit; u is 0 or 1; and -Sulf- is a first spacer unit comprising a sulfatase-cleavable moiety; * denotes a point of attachment to Y; and ## denotes a point of attachment to a drug moiety (-D). For sulfatase-cleavable linkers, see, e.g., Bargh et al., "Sulfatase-cleavable linkers for antibody-drug conjugates", Chemical Science, 2020, 11, 2375-2380, doi: 10.1039 / c9sc06410a.

[0282] Structure*-A a -U AT u -Sulf- ## In a linker having the formula: AT ) (if present) or to the first spacer unit containing the sulfatase-cleavable moiety. The second spacer unit (-A-) serves to link Y to the connecting unit (U AT ). Alternatively, the second spacer unit (-A-) can be any chemical group or moiety that can be linked to a connecting unit (U ATIn the absence of a spacer unit (-Sulf-), Y may be linked to a first spacer unit (-Sulf-) that includes a sulfatase-cleavable moiety. In this regard, Y is attached to a second spacer unit (-A-) as described herein. The second spacer unit (-A-) may be linked to a connecting unit (-U AT Depending on whether -U - AT -) or to a first spacer unit (-Sulf-) that has a sulfatase-cleavable moiety. AT A functional group capable of forming a bond to a sulfatase-cleavable moiety (-Sulf-) or to a first spacer unit containing a sulfatase-cleavable moiety (-Sulf-) is, for example, TIFF2024540691000273.tif12128 or a carbonyl group depicted as -C(O)-. The integer a associated with the second spacer unit can be 0 or 1. Preferably, the integer a is 1. Alternatively, in other embodiments, the second spacer unit is absent (a = 0).

[0283] Linker*-A a -U AT u -Sulf- ## In the formula (I), the second spacer unit -A- can be any second spacer unit as described herein. In some embodiments, the second spacer unit -A-, when present, has the structure: TIFF2024540691000274.tif26128, where TIFF2024540691000275.tif26128 is as defined herein. Thus, in some embodiments, the linker (L) has the structure: TIFF2024540691000276.tif30128, where L P , R S ,s*,M,U AT , u, and Sulf are as defined herein; * denotes the point of attachment to -Y-; and ## denotes the point of attachment to the drug moiety (-D).

[0284] Per connection (-U AT A connecting unit (-U-) may be included when desired to add additional distance between -Y- or, if present, the second spacer unit (-A-) and the first spacer unit containing the sulfatase-cleavable moiety (-Sulf-). AT -), if present, extends the framework of the linker (-L-). In this regard, the linking unit (-U AT -) can be covalently linked at one end to -Y- or, if a second spacer unit -A- is present, to the second spacer unit (-A-), and the connecting unit (-U AT -) is covalently attached at its other end to a first spacer unit (-Sulf-) that contains a sulfatase-cleavable group.

[0285] Per connection (-U AT -) may be any chemical group or moiety that serves to provide a connection of the first spacer unit (-Sulf-), which comprises a sulfatase-cleavable moiety, to the second spacer unit (-A-), if present, or to -Y-.

[0286] In some embodiments, the connection unit (U AT ) has the formula shown below: TIFF2024540691000277.tif22128, wherein v is an integer ranging from 1 to 6; preferably, v is 1 or 2; more preferably, v is 2; and v* is an integer ranging from 1 to 6; preferably, v* is 1 or 2; more preferably, v* is 1; * denotes the point of attachment to the second spacer unit (-A-), if present, and # denotes the point of attachment to the sulfatase-cleavable moiety (Sulf).

[0287] Preferably, the first spacer unit (Sulf) comprising the sulfatase-cleavable moiety has the formula depicted below: TIFF2024540691000278.tif39128 where X is hydrogen (H) or an electron withdrawing group, such as NO 2 * is the unit of connection (U AT ) (if present) or (-A-) (if present); and # denotes the point of attachment to the drug moiety (-D).

[0288] In some embodiments, the linker L can have the structure: TIFF2024540691000279.tif49128 where X is H or NO 2 and; * denotes the point of attachment to -Y-; and ## denotes the point of attachment to the drug moiety (-D).

[0289] In some embodiments, the linker L can have the structure: TIFF2024540691000280.tif52128, where X is H or NO 2 and; R S is a second polyalkylene glycol unit as defined herein; preferably, R S is a second polyethylene glycol unit as defined herein; M is as defined herein; preferably M is -O-; * denotes the point of attachment to -Y-; and ## denotes the point of attachment to the drug moiety (-D).

[0290] Third spacer unit In some embodiments, when the first spacer unit (-B-), or the first spacer unit comprising a sugar moiety (-G-), or the first spacer unit comprising a sulfatase-cleavable moiety (Sulf) is present, the linker (L) may comprise an optional third spacer unit (-E-), which is disposed between the first spacer unit (-B-), or the first spacer unit comprising a sugar moiety (-G-), or the first spacer unit comprising a sulfatase-cleavable moiety (Sulf) and the drug moiety (-D). The third spacer unit may be a functional group that may facilitate the connection of the first spacer unit (-B-), or the first spacer unit comprising a sugar moiety (-G-), or the first spacer unit comprising a sulfatase-cleavable moiety (Sulf) to the drug moiety (-D), or it may provide an additional structural component that may facilitate the release of the drug moiety (-D) from the remainder of the conjugate. Suitable third spacer units are described, for example, in WO 2019 / 236954.

[0291] In some embodiments, the third spacer unit (-E-) is attached to the first spacer unit (-B-) and the drug moiety (-D). Thus, the linker (-L-) has the structure *-A a -W w -B b -E- ## where -E- is a third spacer unit as described herein; and where -A-, a, -W-, w, and -B- can have the structure *-A a -W w -B b - ## is as described herein for linker (L) having the formula:

[0292] In other embodiments, the third spacer unit (-E-) is linked to the first spacer unit (-G-) which comprises a sugar moiety and to the drug moiety (-D-). Thus, the linker (-L-) has the structure *-A a -Q CO q -GE-## where -E- is a third spacer unit as described herein; where -A-, a, -Q CO -, q, and G are specifically in the structure *-A a -Q CO q -G- ## As described herein with respect to linker (L) having the formula:

[0293] In other embodiments, the third spacer unit (-E-) is linked to the first spacer unit (-Sulf-) that comprises a sulfatase-cleavable moiety and to the drug moiety (-D). Thus, the linker (-L-) has the structure *-A a -U AT u -Sulf-E- ## where -E- is a third spacer unit as described herein; where -A-, a, -U AT -, u, and Sulf are specifically in the structure *-A a -U AT u -Sulf- ## As described herein with respect to linker (L) having the formula:

[0294] In some embodiments, an exemplary third spacer unit -E- has the formula: Represented by TIFF2024540691000281.tif55128, In the formula, EWG represents an electron withdrawing group, R 1 is -H or (C 1 -C 4 ) alkyl, and the subscript n is 1 or 2. In some embodiments, the EWG is -CN, -NO 2 , -CX 3 , -X, C(=O)OR', -C(=O)N(R') 2 , -C(=O)R', -C(=O)X, -S(=O)2 R', -S(=O) 2 OR', -S(=O) 2 NHR', -S(=O) 2 N(R') 2 , -P(=O)(OR') 2 , -P(=O)(CH 3 )NHR', -NO, -N(R') 3 + where X is -F, -Br, -Cl, or -I, and R' is selected from the group consisting of hydrogen and (C 1 -C 6 ) alkyl, and where the wavy line adjacent to the nitrogen atom in each of formulas (a), (a'), (a"), (b) and (b') is the point of covalent attachment to the first spacer unit (-B-), and the wavy line adjacent to the carbonyl carbon atom of formula (b) and formula (b') is the point of covalent attachment to a hydroxyl or primary or secondary amine heteroatom of the drug moiety (-D); and where formulas (a), (a') and (a") represent exemplary units, where T* is a heteroatom from a hydroxyl or primary or secondary amine functionality of the drug moiety (-D); and where the wavy line adjacent to T* is the point of covalent attachment to the remainder of the drug moiety. In these embodiments, the third spacer unit -E- may facilitate the release of the drug moiety as a free drug.

[0295] In yet other embodiments, the third spacer unit has the formula: Represented by TIFF2024540691000282.tif38128, wherein each R is independently -H or (C 1 -C 4 Formulas (a1) and (a1') where -CH is an alkyl group represent units where O* is the oxygen atom from a hydroxyl substituent of the drug moiety (-D); and the wavy lines in formulas (a1), (a1') and (b1) retain their previously mentioned meanings from formulas (a), (a') and (b), respectively. In formula (a1'), -CH 2 CH 2 N + (R) 2The moiety represents an exemplary basic unit in protonated form.

[0296] Drug Moiety (-D) The present disclosure provides conjugates, such as antibody-drug conjugates, that include a drug moiety. The terms "drug moiety" or "payload", which may be used interchangeably, as used herein refer to a chemical or biochemical moiety that is conjugated to a receptor binding molecule (RBM), such as an antibody or an antigen-binding fragment. In this regard, reference is again made to the conjugate of formula (I) described herein. The receptor binding molecule (RBM) can be conjugated to several identical or different drug moieties using any method described herein or known in the art. In some embodiments, the drug moiety can be a molecule that has a cytotoxic effect on mammalian cells, can cause apoptosis, and / or can have a regulatory effect on malignant cells. The drug moiety can be hydrophobic.

[0297] In some preferred embodiments, the drug moiety is an anti-cancer drug.Thus, the drug may be selected from the group consisting of maytansinoids, calicheamicins, tubulysins, amatoxins, dolastatins, and auristatins such as monomethylauristatin E (MMAE) or monomethylauristatin F (MMAF), pyrrolobenzodiazepine dimers, indolino-benzodiazepine dimers, emetines, radioisotopes, therapeutic proteins and peptides (or fragments thereof), kinase inhibitors, CDK inhibitors, histone deacetylase (HDAC) inhibitors, MEK inhibitors, KSP inhibitors, and analogs or prodrugs thereof.In preferred embodiments, the drug moiety is MMAE or MMAF.More preferably, the drug moiety is MMAE.

[0298] In some embodiments, the drug moiety is a maytansinoid drug moiety and has the structure: including one with TIFF2024540691000283.tif51128, where the wavy line indicates the covalent attachment of the sulfur atom of the maytansinoid to a linker of a conjugate, such as an antibody-drug conjugate. R at each occurrence is independently H or C1-C6 alkyl. The alkylene chain connecting the amide group to the sulfur atom can be methanyl, ethanyl, propanyl, i.e., m is 1, 2, or 3 (U.S. Patent No. 633,410, U.S. Patent No. 5,208,020, Chari et al. (1992) Cancer Res. 52; 127-131, Lui et al. (1996) Proc. Natl. Acad. Sci. 93:8618-8623).

[0299] All stereoisomers of the maytansinoid drug moiety, i.e., any combination of R and S configurations at the chiral carbon of the maytansinoid, are contemplated for the conjugates disclosed herein. In some embodiments, the maytansinoid drug moiety has the following stereochemistry: I have TIFF2024540691000284.tif52128.

[0300] In some embodiments, the maytansinoid drug moiety is N 2' -Deacetyl-N 2' -(3-mercapto-1-oxopropyl)-maytansine (also known as DM1). DM1 has the following structural structure: Represented by TIFF2024540691000285.tif52128.

[0301] In some embodiments, the maytansinoid drug moiety is N 2' -Deacetyl-N 2' -(4-mercapto-1-oxopentyl)-maytansine (also known as DM3). DM3 has the following structural structure: Represented by TIFF2024540691000286.tif59128.

[0302] In some embodiments, the maytansinoid drug moiety is N 2' -Deacetyl-N2' -(4-methyl-4-mercapto-1-oxopentyl)-maytansine (also known as DM4). DM4 has the following structural structure: Represented by TIFF2024540691000287.tif48128.

[0303] Preferably, in the conjugates disclosed herein that include a maytansinoid drug moiety, the maytansinoid is N 2' -Deacetyl-N 2' -(3-mercapto-1-oxopropyl)-maytansine (DM1) or N 2' -Deacetyl-N 2’ -(4-mercapto-4-methyl-1-oxopentyl)-maytansine (DM4).

[0304] The drug moiety may be a calicheamicin. "Calicheamicin," as used herein, refers to a class of enediyne antitumor antibiotics derived from the bacterium Micromonospora echinospora, most notably calicheamicin γ1. It was originally isolated in the mid-1980s from the chalk earth, or "Calich Pits," in Kerrville, Texas. It is highly toxic to all cells. Thus, the drug moiety may be substituted or derivatized for coupling to a linker and / or receptor binding molecule, with the following structure: It may be calicheamicin gamma 1, exemplified by TIFF2024540691000288.tif45128.

[0305] The drug moiety can be tubulysin. Tubulysin has functions as an anti-microtubule agent, anti-mitotic agent, apoptosis inducer, anti-cancer agent, anti-angiogenic agent, and anti-proliferative agent. Tubulysin is a cytotoxic peptide that contains nine members (A-I). Preferably, the tubulysin is tubulysin A. Tubulysin A has potential application as an anti-cancer agent. It arrests cells in the G2 / M phase. Tubulysin A has the following structure: I have TIFF2024540691000289.tif41128.

[0306] The drug moiety may be an amatoxin. Amatoxins are a collective term for a subgroup of at least eight related toxic compounds found in several genera of poisonous mushrooms, notably Amanita phalloides and several other members of the Amanita genus, as well as some Conocybe, Galerina, and Lepiota mushroom species. Small amounts of amatoxins are lethal. These compounds have a similar structure, that of eight amino acid residues arranged in a conserved macrobicyclic motif (an overall pentacyclic structure if one counts the rings intrinsic to the proline and tryptophan derived residues). All amatoxins are oligopeptides, synthesized as a 35 amino acid proprotein from which the last eight amino acids are cleaved by prolyl oligopeptidase. The general amino acid sequence of amatoxins is Ile-Trp-Gly-Ile-Gly-Cys-Asn-Pro, with a bridge between Trp and Cys via a sulfoxide (S=O) moiety and a hydroxylation in variants of the molecule. Currently, there are 10 known amatoxins, which can be drug moieties. TIFF2024540691000290.tif136128

[0307] The drug moiety can be a dolastatin, such as dolastatin 10 or dolastatin 15. Both are marine natural products isolated from the Indian Ocean sea hare Dollabella auricularia. This potent antitumor agent has also been isolated from the Palauan marine cyanobacterium Symploca sp. VP642. Dolastatins 10 and 15, small linear peptide molecules, are considered anticancer drugs that show efficacy against breast cancer, liver cancer, solid tumors, and some leukemias. Preclinical studies have shown efficacy in experimental antineoplastic and tubulin assembly systems. Dolastatins are mitotic inhibitors. They inhibit microtubule assembly by interfering with the formation of tubulin, thereby interfering with cell division by mitosis and inducing apoptosis and Bcl-2 phosphorylation in some malignant cell types. Dolostatin 10 (N,N-dimethyl-L-valyl-N-[(3R,4S,5S)-3-methoxy-1-{(2S)-2-[(1R,2R)-1-methoxy-2-methyl-3-oxo-3-{[(1S)-2-phenyl-1-(1,3-thiazol-2-yl)ethyl]amino}propyl]-1-pyrrolidinyl}-5-methyl-1-oxo-4-heptanyl]-N-methyl-L-valinamide) has the following structure: I have TIFF2024540691000291.tif45128.

[0308] Dolastatin 15 ((2S)-1-[(2S)-2-benzyl-3-methoxy-5-oxo-2,5-dihydro-1H-pyrrol-1-yl]-3-methyl-1-oxo-2-butanyl N,N-dimethyl-L-valyl-L-valyl-N-methyl-L-valyl-L-prolyl-L-prol) has the following structure: I have TIFF2024540691000292.tif51128.

[0309] In a preferred embodiment of the conjugates, e.g., antibody drug conjugates, disclosed herein, the drug moiety is an auristatin. Preferably, the auristatin is monomethylauristatin F (MMAF) or monomethylauristatin E (MMAE). More preferably, the auristatin is monomethylauristatin E (MMAE).

[0310] In some embodiments of the antibody drug conjugates described herein, the drug moiety is monomethylauristatin F (also known as MMAF). MMAF has the following structural formula: It is represented by TIFF2024540691000293.tif34128. Preferably, MMAF is attached to the linker L via its N-terminus, indicated by an asterisk ("*").

[0311] In some embodiments, the auristatin drug moiety is monomethylauristatin E (also known as MMAE), which has the following structural formula: Represented by TIFF2024540691000294.tif38128. Preferably, MMAE is attached to the linker L via its N-terminus, indicated by an asterisk ("*").

[0312] These molecules have been shown to non-competitively inhibit the binding of vincristine to tubulin (at a location known as the vinca / peptide domain), but bind to the RZX / MAY domain.

[0313] The drug moiety may be optionally substituted or derivatized for coupling to the linker and / or receptor binding molecule, and has the following structure: The compound may be a pyrrolobenzodiazepine dimer, such as the compound having TIFF2024540691000295.tif20128.

[0314] The drug moiety has the following structure: The compound may be an indolinobenzodiazepine dimer, such as the compound having TIFF2024540691000296.tif29128.

[0315] The drug moiety can be emetine. Emetine exerts antitumor effects by apoptosis through mechanisms such as inhibition of protein biosynthesis, DNA interaction, and modulation of proapoptotic factors (see, e.g., Uzor, "Recent Developments on Potential New Applications of Emetine as Anti-Cancer Agent", EXCLI Journal 2016; 15:323-238, http: / / dx.doi.org / 10.17179 / excli2016-280). Emetine has the following structure: TIFF2024540691000297.tif69128. Emetine can be attached to the linker L via the nitrogen atom marked with an asterisk ("*").

[0316] The drug moiety can be a radioisotope. Exemplary radioisotopes described herein can be associated with brachytherapy, usually a gamma or beta emitter, such as iodine-125, iodine-131, iridium-192, or palladium-103.

[0317] The drug moiety may be a therapeutic protein or peptide or a fragment thereof. Typical examples are cytokines such as interleukins, ricin, diphtheria toxin, Pseudomonas aeruginosa exotoxin PE38, etc.

[0318] The drug moiety may be a kinase inhibitor, preferably an inhibitor of a kinase associated with a pro-tumorigenic function. Exemplary kinase inhibitors include imatinib, nilotinib, dasatinib, bosutinib, ponatinib, gefitinib, erlotinib, afatinib, osimertinib, lapatinib, neratinib, sorafenib, sunitinib, pazopanib, axitinib, lenvatinib, cabozatinib, vandetanib, regorafenib, vemurafenib, dabrafenib, trametinib, cobimetinib, crizotinib, certinib, alectinib, brigatinib, loratinib, ibrutinib, acalibrutinib, midostaurin, ruxolitinib, idelalisib, copanlisib, palbociclib, ribociclib, or abemaciclib.

[0319] The drug moiety may be a CDK (cyclin-dependent kinase) inhibitor. "CDK inhibitor" as used herein means any chemical or drug that inhibits the function of a CDK (cyclin-dependent kinase). A CDK inhibitor that may be used as the drug moiety in the present disclosure is AT7519 (see, e.g., Santo et al., AT7519, "A novel small molecule multi-cyclin-dependent kinase inhibitor, induces apoptosis in multiple myeloma via GSK-3beta activation and RNA polymerase II inhibition", Oncogene (2010) 29, 2325-2336, doi: 10.1038 / onc.2009.510). AT7519 has the following structure: TIFF2024540691000298.tif50128. AT7519 can be attached to the linker L via the nitrogen atom marked with an asterisk ("*").

[0320] The drug moiety can be a histone deacetylase (HDAC) inhibitor. A histone deacetylase inhibitor is a compound that inhibits histone deacetylase (HDAC). A histone deacetylase inhibitor that can be used as a drug moiety in the present disclosure is panobinostat (see, for example, Rasmussen et al., "Panobinostat, a histone deacetylase inhibitor, for latent-virus reactivation in HIV-infected patients on suppressive antiretroviral therapy: a phase 1 / 2, single groups, clinical trial", Lancet HIV 2014, 1:e13-21, http: / / dx.doi.org / 10.1016 / S2352-3018(14)70014-1). Panobinostat has the following structure: TIFF2024540691000299.tif95128. AT7519 can be attached to the linker L via the nitrogen atom marked with an asterisk ("*").

[0321] The drug moiety can be a MEK inhibitor. MEK inhibitors, as used herein, refer to chemicals or drugs that inhibit the mitogen-activated protein kinase kinase enzymes MEK1 and / or MEK2. They can be used to affect the MAPK / ERK pathway, which is often overactive in some cancers. Thus, MEK inhibitors are promising for the treatment of some cancers, especially BRAF-mutated melanoma, and KRAS / BRAF-mutated colorectal cancer. Exemplary MEK inhibitors include trametinib (GSK1120212), cobimetinib or XL518, binimetinib (MEK162), selumetinib, PD-325901, CI-1040, PD035901, or TAK-733.

[0322] The drug moiety can be a KSP (kinesin spindle protein) inhibitor. Examples of KSP inhibitors include ispinesib (SB-715992), SB743921, AZ 3146, GSK923295, BAY 1217389, MPI-0479605, and ARQ 621. In some embodiments, SB743921 can be used as a drug moiety in the present disclosure (see, for example, Song et al., "KSP inhibitor SB743921 induces death of multiple myeloma cells via inhibition of the NF-kB signaling pathway", BMB Reports 2015, 48(10): 571-576, http: / / dx.doi.org / 10.5483 / BMBRep.2015.48.10.015). SB743921 has the following structure: TIFF2024540691000300.tif72128. SB743921 can be attached to the linker L via the nitrogen atom marked with an asterisk ("*").

[0323] The present invention also relates to a conjugate having formula (I): TIFF2024540691000301.tif34128, or a pharma- ceutically acceptable salt or solvate thereof; During the ceremony: RBM is an antibody; TIFF2024540691000302.tif6128 is a double bond; or TIFF2024540691000303.tif6128 is a single bond; V is not present if TIFF2024540691000304.tif6128 is a double bond; or V is TIFF2024540691000305.tif6128 is a single bond, H; X is R if TIFF2024540691000306.tif6128 is a double bond 3 -C; or X is If TIFF2024540691000307.tif6128 is a single bond, TIFF2024540691000308.tif9128; Y is NH; R 1 The structure: TIFF2024540691000309.tif15128, Where: TIFF2024540691000310.tif9128 shows the position of O; K F is as defined herein; preferably K F is H; and o is an integer as defined herein; Preferably, o is an integer ranging from 8 to 30; More preferably, o is an integer ranging from 4 to 16; even more preferably, o is an integer ranging from 8 to 16; even more preferably, o is 10, 11, 12, 13 or 14; even more preferably, o is 11, 12 or 13; still more preferably, o is 12; or More preferably, o is an integer ranging from 16 to 30; even more preferably, o is an integer ranging from 20 to 28; even more preferably, o is 22, 23, 24, 25 or 26; even more preferably, o is 23, 24 or 25; still more preferably, o is 24; R 3 is H; R 4 is H; L has the following structure: TIFF2024540691000311.tif43128, where # indicates the point of attachment to Y and * indicates the point of attachment to the drug moiety (D); D is a drug moiety; m is 1; and n is an integer as defined herein; Preferably, n is an integer ranging from 1 to 10; more preferably from 2 to 10; even more preferably from 4 to 10; even more preferably from 6 to 10, even more preferably from 7 to 10, still more preferably n is 8; or Preferably, n is an integer ranging from 1 to 10, more preferably from 2 to 8, even more preferably from 3 to 6, even more preferably n is 4 or 5, and even more preferably n is 4.

[0324] Preferably, drug moiety D is monomethylauristatin E (MMAE) or monomethylauristatin F (MMAF).

[0325] More preferably, the drug moiety D is monomethylauristatin E (MMAE).

[0326] The present invention also relates to a conjugate having the following formula (Ia): TIFF2024540691000312.tif64148 in formula: The RBM is an antibody; and n is an integer as defined herein; Preferably, n is an integer ranging from 1 to 10; more preferably from 2 to 10; even more preferably from 4 to 10; even more preferably from 6 to 10, even more preferably from 7 to 10, still more preferably n is 8; or Preferably, n is an integer ranging from 1 to 10, more preferably from 2 to 8, even more preferably from 3 to 6, even more preferably n is 4 or 5, and even more preferably n is 4.

[0327] The present invention also relates to a conjugate having the following formula (Ib): TIFF2024540691000313.tif65148 formula: The RBM is an antibody; and n is an integer as defined herein; preferably n is an integer ranging from 1 to 10; more preferably from 2 to 10; even more preferably from 4 to 10; even more preferably from 6 to 10, even more preferably from 7 to 10, still more preferably n is 8; or Preferably, n is an integer ranging from 1 to 10, more preferably from 2 to 8, even more preferably from 3 to 6, even more preferably n is 4 or 5, and even more preferably n is 4.

[0328] Compound of formula (II) The present invention also relates to a compound having the formula (II): TIFF2024540691000314.tif29128, or a pharma- ceutically acceptable salt or solvate thereof, During the ceremony: TIFF2024540691000315.tif6128 is a triple bond; or TIFF2024540691000316.tif6128 is a double bond; V is absent if TIFF2024540691000317.tif6128 is a triple bond; or V is If TIFF2024540691000318.tif6128 is a double bond, H or (C 1 -C 8 ) alkyl; X is R if TIFF2024540691000319.tif6128 is a triple bond 3 -C: or X is If TIFF2024540691000320.tif6128 is a double bond, TIFF2024540691000321.tif9128; Y is NR 5 , S, O, or CR 6 R 7 and; R 1is a first polyalkylene glycol unit R comprising at least three alkylene glycol subunits; F and; R 3 is H; or an optionally substituted aliphatic or an optionally substituted aromatic residue; R 4 is H; or an optionally substituted aliphatic or an optionally substituted aromatic residue; R 5 is H; or an optionally substituted aliphatic or an optionally substituted aromatic residue; R 6 is H; or an optionally substituted aliphatic or an optionally substituted aromatic residue; R 7 is H; or an optionally substituted aliphatic or an optionally substituted aromatic residue; L is a linker; D is a drug moiety; and m is an integer ranging from 1 to 10.

[0329] Preferably R 3 is H or (C 1 -C 8 ) alkyl; more preferably R 3 is H. Preferably, R 4 If present, H or (C 1 -C 8 ) alkyl; more preferably R 4 When present, it is H. Preferably, R 5 If present, H or (C 1 -C 8 ) alkyl; more preferably R 5 When present, it is H. Preferably, R 6 If present, H or (C 1 -C 8 ) alkyl; more preferably R 6 When present, it is H. Preferably, R 7 If present, H or (C 1 -C 8) alkyl; more preferably R 7 is H, if present.

[0330] Preferably, TIFF2024540691000322.tif6128 is a triple bond; V is absent; X is R 3 -C; and R 3 is H or an optionally substituted aliphatic or an optionally substituted aromatic residue; preferably R 3 is H or (C 1 -C 8 ) alkyl; more preferably R 3 is H.

[0331] More preferably, TIFF2024540691000323.tif6128 represents a triple bond; V is absent; X is R 3 -C and R 3 is H or (C 1 -C 8 ) alkyl. Preferably, R 3 is H or (C 1 -C 6 ) alkyl, more preferably H or (C 1 -C 4 ) alkyl, even more preferably H or (C 1 -C 2 ) alkyl. Even more preferably, R 3 is H.

[0332] In some embodiments, TIFF2024540691000324.tif6128 can be a double bond; V can be H or (C 1 -C 8 ) alkyl, preferably V is H; X is TIFF2024540691000325.tif9128;R 3 is H or an optionally substituted aliphatic or an optionally substituted aromatic residue; more preferably R 3 is H or (C 1-C 8 ) alkyl, more preferably R 3 is H;R 4 is H or an optionally substituted aliphatic or an optionally substituted aromatic residue; preferably, R 4 is H or (C 1 -C 8 ) alkyl, preferably R 4 is H.

[0333] In some embodiments, TIFF2024540691000326.tif6128 may represent a double bond; V may represent H or (C 1 -C 8 ) alkyl; X can be Represents TIFF2024540691000327.tif9128; and R 3 and R 4 are independently H or (C 1 -C 8 ) alkyl. Preferably, R 3 and R 4 are independently H or (C 1 -C 6 ) alkyl, more preferably H or (C 1 -C 4 ) alkyl, even more preferably H or (C 1 -C 2 ) alkyl. Preferably, R 3 and R 4 are the same; even more preferably, R 3 , R 4 and V are the same. More preferably, R 3 and R 4 are both H. Preferably, V is H or (C 1 -C 6 ) alkyl, more preferably H or (C 1 -C 4 ) alkyl, even more preferably H or (C 1 -C 2 ) alkyl. Even more preferably, V is H. In a preferred embodiment, R 3 , R4 and V are each H.

[0334] In any one of the compounds of formula (II), the variables may be defined as described herein, particularly with respect to the conjugate of formula (I) and / or the thiol-containing molecule of formula (III). Thus, RBM, TIFF2024540691000328.tif6128, V, X, Y, R 1 , R 3 , R 4 , R 5 , R 6 , R 7 , L, D, m and n are as defined herein. Preferably, Y is NH.

[0335] Methods for preparing conjugates of formula (I) The present invention also relates to a method for preparing a conjugate of formula (I), the method comprising the steps of: Compounds of formula (II): TIFF2024540691000329.tif29128, or a pharma- ceutically acceptable salt or solvate thereof, During the ceremony: TIFF2024540691000330.tif6128 is a triple bond; or TIFF2024540691000331.tif6128 is a double bond; V is absent if TIFF2024540691000332.tif6128 is a triple bond; or V is If TIFF2024540691000333.tif6128 is a double bond, H or (C 1 -C 8 ) alkyl; X is R if TIFF2024540691000334.tif6128 is a triple bond 3 -C: or X is If TIFF2024540691000335.tif6128 is a double bond, TIFF2024540691000336.tif9128; Y is NR 5 , S, O, or CR 6 R 7 and; R 1 is a first polyalkylene glycol unit R comprising at least three alkylene glycol subunits; F and; R 3 is H; or an optionally substituted aliphatic or an optionally substituted aromatic residue; R 4 is H; or an optionally substituted aliphatic or an optionally substituted aromatic residue; R 5 is H; or an optionally substituted aliphatic or an optionally substituted aromatic residue; R 6 is H; or an optionally substituted aliphatic or an optionally substituted aromatic residue; R 7 is H; or an optionally substituted aliphatic or an optionally substituted aromatic residue; L is a linker; D is a drug moiety; and m is an integer ranging from 1 to 10; A compound of formula (II) or a pharma- ceutically acceptable salt or solvate thereof, A thiol-containing molecule of formula (III): TIFF2024540691000337.tif9128 formula, RBM is a receptor binding molecule; and n is an integer ranging from 1 to 20; a thiol-containing molecule of formula (III) By reacting Compounds of formula (I): TIFF2024540691000338.tif35128, or a pharma- ceutically acceptable salt or solvate thereof, During the ceremony: TIFF2024540691000339.tif5128 is a compound of formula (II) TIFF2024540691000340.tif6128 is a triple bond if it is a double bond; or TIFF2024540691000341.tif5128 is a compound of formula (II) TIFF2024540691000342.tif6128 is a double bond, then it is a single bond; V is not present if TIFF2024540691000343.tif5128 is a double bond; or V is If TIFF2024540691000344.tif5128 is a single bond, it is H or (C 1 -C 8 ) alkyl; X is R if TIFF2024540691000345.tif6128 is a double bond 3 -C; or X is If TIFF2024540691000346.tif6128 is a single bond, TIFF2024540691000347.tif9128; Y is NR 5 , S, O, or CR 6 R 7 and; R 1 is a first polyalkylene glycol unit R comprising at least three alkylene glycol subunits; F and; R 3 is H; or an optionally substituted aliphatic or an optionally substituted aromatic residue; R 4 is H; or an optionally substituted aliphatic or an optionally substituted aromatic residue; R 5 is H; or an optionally substituted aliphatic or an optionally substituted aromatic residue; R 6 is H; or an optionally substituted aliphatic or an optionally substituted aromatic residue; R 7 is H; or an optionally substituted aliphatic or an optionally substituted aromatic residue; L is a linker; D is a drug moiety; m is an integer ranging from 1 to 10; and n is an integer ranging from 1 to 20; A compound of formula (I) or a pharma- ceutically acceptable salt or solvate thereof The method includes the step of generating

[0336] Preferably R 3 is H or (C 1 -C 8 ) alkyl; more preferably R 3 is H. Preferably, R 4 If present, H or (C 1 -C 8 ) alkyl; more preferably R 4 When present, it is H. Preferably, R 5 If present, H or (C 1 -C 8 ) alkyl; more preferably R 5 When present, it is H. Preferably, R 6 If present, H or (C 1 -C 8 ) alkyl; more preferably R 6 When present, it is H. Preferably, R 7 If present, H or (C 1 -C 8 ) alkyl; more preferably R 7 is H, if present.

[0337] Preferably, TIFF2024540691000348.tif6128 is a triple bond; V is absent; X is R 3 -C; and R3 is H or an optionally substituted aliphatic or an optionally substituted aromatic residue; preferably R 3 is H or (C 1 -C 8 ) alkyl; more preferably R 3 is H; and TIFF2024540691000349.tif6128 represents a double bond.

[0338] More preferably, TIFF2024540691000350.tif6128 represents a triple bond; V is absent; X is R 3 -C stands for R 3 is H or (C 1 -C 8 ) alkyl; and TIFF2024540691000351.tif6128 represents a double bond. 3 is H or (C 1 -C 6 ) alkyl, more preferably H or (C 1 -C 4 ) alkyl, even more preferably H or (C 1 -C 2 ) alkyl. Even more preferably, R 3 is H.

[0339] In some embodiments, TIFF2024540691000352.tif6128 can be a double bond; V can be H or (C 1 -C 8 ) alkyl, preferably V is H; X is TIFF2024540691000353.tif9128;R 3 is H or an optionally substituted aliphatic or an optionally substituted aromatic residue; and TIFF2024540691000354.tif6128 may represent a bond; more preferably R 3 is H or (C 1 -C 8) alkyl, more preferably R 3 is H;R 4 is H or an optionally substituted aliphatic or an optionally substituted aromatic residue; preferably, R 4 is H or (C 1 -C 8 ) alkyl, preferably R 4 is H.

[0340] In some embodiments, TIFF2024540691000355.tif6128 may represent a double bond; V may represent H or (C 1 -C 8 ) alkyl; X can be Represents TIFF2024540691000356.tif9128;R 3 and R 4 are independently H or (C 1 -C 8 ) alkyl; and TIFF2024540691000357.tif6128 may represent a bond. 3 and R 4 are independently H or C 1 -C 6 -alkyl, more preferably H or C 1 -C 4 -alkyl, even more preferably H or C 1 -C 2 -alkyl. Preferably, R 3 and R 4 are the same; even more preferably, R 3 , R 4 and V are the same. More preferably, R 3 and R 4 are both H. Preferably, V is H or C 1 -C 6 -alkyl, more preferably H or C 1 -C 4 -alkyl, even more preferably H or C 1 -C 2-alkyl. Even more preferably, V is H. In a preferred embodiment, R 3 , R 4 and V are each H.

[0341] Expressions used in this specification TIFF2024540691000358.tif6128 and With respect to TIFF2024540691000359.tif6128, it should be noted that, as is commonly known to those skilled in the art, each carbon atom is tetravalent. Thus, the structure: TIFF2024540691000360.tif18128, where X and V are as defined herein and an asterisk (*) indicates the connection to phosphorus, has the structure: TIFF2024540691000361.tif23128 (where R 3 , R 4 and V is as defined herein. TIFF2024540691000362.tif19128, where X and V are as defined herein, the asterisk (*) indicates the connection to phosphorus, and the # indicates the connection to the receptor binding molecule (RBM), has the structure: TIFF2024540691000363.tif22128 (where R 3 , R 4 and V is as defined herein, and H is hydrogen. The wavy bond indicates that the configuration of the double bond may be E or Z. It is also possible that the compounds exist as a mixture of E and Z isomers.

[0342] If the receptor-binding molecule, such as an antibody, contains one or more disulfide bridges, the method may further comprise reducing at least one disulfide bridge of the receptor-binding molecule in the presence of a reducing agent to form a thiol group (SH). The resulting compound of formula (III) may then be reacted with a compound of formula (II) to produce a conjugate of formula (I). The reducing agent may be selected from the group consisting of tris(2-carboxyethyl)phosphine (TCEP), dithiothreitol (DTT), sodium dithionite, sodium thiosulfate, and sodium sulfite. Thus, the reducing agent may be dithiothreitol (DTT). The reducing agent may be sodium dithionite. The reducing agent may be sodium sulfite. Preferably, the reducing agent is tris(2-carboxyethyl)phosphine (TCEP).

[0343] Preferably, the reduction of at least one disulfide bridge comprises using about 1 to about 3 equivalents, preferably about 1 to about 2 equivalents, more preferably about 1 equivalent of reducing agent per disulfide bridge to be reduced. In this context, it is noted that theoretically, one equivalent of reducing agent, in particular a reducing agent as described herein, is required to reduce one disulfide bridge to obtain two thiol groups (SH).

[0344] Preferably, the thiol-containing molecule of formula (III) is reacted with about 1 to about 4 equivalents, preferably about 1 to about 3 equivalents, more preferably about 1 to about 2 equivalents, and even more preferably about 1.5 equivalents of the compound of formula (II) per thiol group (SH).

[0345] Preferably, the reaction of the compound of formula (II) with the thiol-containing molecule of formula (III) is carried out in an aqueous medium.

[0346] Preferably, the reaction of the compound of formula (II) with the thiol-containing molecule of formula (III) is carried out under neutral pH or slightly basic conditions. Even more preferably, the reaction is carried out at a pH between 6 and 10. Even more preferably, the reaction is carried out at a pH between 7 and 9.

[0347] In any one of the methods, the variables may be defined as described herein, particularly with respect to the conjugate of formula (I) and / or the compound of formula (II). Thus, RBM, TIFF2024540691000364.tif9128, V, X, Y, R 1 , R 3 , R 4 , R 5 , R 6 , R 7 , L, D, m and n are as defined herein. Preferably, Y is NH.

[0348] Methods for preparing compounds of formula (II) are known in the art. As an illustrative example, compounds of formula (II) in which the group Y is NH can be prepared, for example, by using techniques and conditions such as those described in WO 2018 / 041985 A1, which is incorporated herein by reference, for example, the Staudinger phosphonite reaction. Compounds of formula (II) in which Y is S or O can be prepared, for example, by using techniques and conditions such as those described in WO 2019 / 170710, which is incorporated herein by reference. Compounds of formula (I) in which Y is S or O can be prepared, for example, by using techniques and conditions such as those described in WO 2019 / 170710, which is incorporated herein by reference, for example, by using techniques and conditions such as those described in WO 2019 / 170710, which is incorporated herein by reference, for example, by using techniques and conditions such as those described in WO 2019 / 170710, which is incorporated herein by reference, for example, by using techniques and conditions such as those described in WO 2019 / 170710, which is incorporated herein by reference, for example, by using techniques and conditions such as those described in WO 2019 / 170710, which is incorporated herein by reference, for example, by using techniques and conditions such as those described in WO 2019 / 170710, which are ... 6 R 7 Compounds of formula (II) may be prepared by substituting at the phosphorus atom, for example, with a suitable organometallic compound, such as a Grignard compound or an organolithium compound, as illustrative examples. Those skilled in the art will readily select suitable methods and conditions for preparing compounds of formula (II). The Examples section of the present disclosure also includes guidance on how to prepare or obtain compounds of formula (II) and / or conjugates of formula (I).

[0349] The present invention also relates to a conjugate of formula (I) obtainable or obtained by any process for preparing a conjugate of formula (I) as described herein.

[0350] Pharmaceutical Compositions The present invention further relates to a pharmaceutical composition comprising the conjugate of formula (I).

[0351] The pharmaceutical composition may comprise a population of conjugates of formula (I), wherein the average number of drug moieties D per receptor-binding molecule in the composition is greater than 0 to about 14, preferably about 1 to about 14, more preferably about 2 to about 14, even more preferably about 4 to about 14, even more preferably about 5 to about 12, even more preferably about 6 to about 12, even more preferably about 7 to about 10, and even more preferably about 8. Thus, the pharmaceutical composition may comprise a population of conjugates of formula (I), wherein the average number of drug moieties D per receptor-binding molecule in the composition is greater than 0 to about 14. Preferably, the pharmaceutical composition comprises a population of conjugates of formula (I), wherein the average number of drug moieties D per receptor-binding molecule in the composition is greater than 0 to about 14. More preferably, the pharmaceutical composition comprises a population of conjugates of formula (I), wherein the average number of drug moieties D per receptor-binding molecule in the composition is greater than 2 to about 14. Even more preferably, the pharmaceutical composition comprises a population of conjugates of formula (I), wherein the average number of drug moieties D per receptor-binding molecule in the composition is about 4 to about 14. Even more preferably, the pharmaceutical composition comprises a population of conjugates of formula (I), wherein the average number of drug moieties D per receptor-binding molecule in the composition is about 5 to about 12. Even more preferably, the pharmaceutical composition comprises a population of conjugates of formula (I), wherein the average number of drug moieties D per receptor-binding molecule in the composition is about 6 to about 12. Even more preferably, the pharmaceutical composition comprises a population of conjugates of formula (I), wherein the average number of drug moieties D per receptor-binding molecule in the composition is about 6 to about 10. Even more preferably, the pharmaceutical composition comprises a population of conjugates of formula (I), wherein the average number of drug moieties D per receptor-binding molecule in the composition is about 8. When the receptor-binding molecule is an antibody or antibody fragment in some preferred embodiments, such average number is also referred to as the "average drug-antibody ratio (DARav)".In this context, one of skill in the art will understand that a composition may comprise a population of conjugates that may vary in the number of drug moieties per receptor-binding molecule and may optionally include unconjugated receptor-binding molecules, resulting in an average number of drug moieties per receptor-binding molecule.

[0352] The pharmaceutical composition may comprise a population of conjugates of formula (I), wherein the average number of drug moieties D per receptor-binding molecule is greater than 0 to about 14, preferably about 1 to about 14, more preferably about 1 to about 12, even more preferably about 2 to about 10, even more preferably about 2 to about 8, even more preferably about 2 to about 6, even more preferably about 3 to about 5, and even more preferably about 4. Thus, the pharmaceutical composition may comprise a population of conjugates of formula (I), wherein the average number of drug moieties D per receptor-binding molecule is greater than 0 to about 14. Preferably, the pharmaceutical composition comprises a population of conjugates of formula (I), wherein the average number of drug moieties D per receptor-binding molecule is greater than 0 to about 14. More preferably, the pharmaceutical composition comprises a population of conjugates of formula (I), wherein the average number of drug moieties D per receptor-binding molecule is greater than 1 to about 14. Even more preferably, the pharmaceutical composition comprises a population of conjugates of formula (I), wherein the average number of drug moieties D per receptor-binding molecule is about 2 to about 10. Even more preferably, the pharmaceutical composition comprises a population of conjugates of formula (I), wherein the average number of drug moieties D per receptor-binding molecule is about 2 to about 8. Even more preferably, the pharmaceutical composition comprises a population of conjugates of formula (I), wherein the average number of drug moieties D per receptor-binding molecule is about 2 to about 6. Even more preferably, the pharmaceutical composition com...

Claims

1. A conjugate having formula (I): or a pharmaceutically acceptable salt or solvate thereof, During the ceremony: RBM is a receptor binding molecule; is a double bond; or is a single bond; V is is absent when is a double bond; or V is If is a single bond, H or (C 1 -C 8 ) alkyl; X is If is a double bond, R 3 -C; or X is If is a single bond, and Y is NR 5 , S, O, or CR 6 R 7 and R 1 is a first polyalkylene glycol unit R comprising at least three alkylene glycol subunits F and R 3 is H; or an optionally substituted aliphatic residue or an optionally substituted aromatic residue; R 4 is H; or an optionally substituted aliphatic residue or an optionally substituted aromatic residue; R 5 is H; or an optionally substituted aliphatic residue or an optionally substituted aromatic residue; R 6 is H; or an optionally substituted aliphatic residue or an optionally substituted aromatic residue; R 7 is H; or an optionally substituted aliphatic residue or an optionally substituted aromatic residue; L is a linker; D is a drug moiety; m is an integer ranging from 1 to 10; and n is an integer ranging from 1 to 20; The conjugate, or a pharmaceutically acceptable salt or solvate thereof.

2. is a double bond; V is absent; X is R 3 -C; and R 3 is H or an optionally substituted aliphatic residue or an optionally substituted aromatic residue; preferably R 3 is H or (C 1 -C 8 ) alkyl; more preferably R 3 The conjugate of claim 1, wherein

3. 2. The conjugate of claim 1, wherein the receptor-binding molecule is selected from the group consisting of an antibody, an antibody fragment, and a proteinaceous binding molecule with antibody-like binding properties.

4. 4. The conjugate of claim 3, wherein the receptor-binding molecule is an antibody.

5. 5. The conjugate of claim 4, wherein the antibody is selected from the group consisting of a monoclonal antibody, a chimeric antibody, a humanized antibody, a human antibody, and a single domain antibody, such as a camel or shark single domain antibody.

6. 2. The conjugate of claim 1, wherein Y is NH.

7. 7. The conjugate of claim 6, wherein the receptor-binding molecule is an antibody.

8. First polyalkylene glycol unit R F But the structure:

2. The conjugate of claim 1, comprising 3 to 100 subunits having the formula:

9. R F but, and where: indicates the position of O; K F -H, -PO 3 H, -(C 1 -C 10 ) alkyl, -(C 1 -C 10 )Alkyl-SO 3 H, -(C 2 -C 10 )Alkyl-CO 2 H, -(C 2 -C 10 ) alkyl-OH, -(C 2 -C 10 )Alkyl-NH 2 , -(C 2 -C 10 ) alkyl-NH(C 1 -C 3 ) alkyl and -(C 2 -C 10 ) alkyl-N((C 1 -C 3 )Alkyl) 2 and 9. The conjugate of claim 8, wherein o is an integer ranging from 3 to 100.

10. R F is a first polyethylene glycol unit comprising at least three ethylene glycol subunits.

11. First polyethylene glycol unit R F But the structure:

11. The conjugate of claim 10, comprising 3 to 100 subunits having the formula:

12. R F but, and where: indicates the position of O; K F -H, -PO 3 H, -(C 1 -C 10 ) alkyl, -(C 1 -C 10 )Alkyl-SO 3 H, -(C 2 -C 10 )Alkyl-CO 2 H, -(C 2 -C 10 ) alkyl-OH, -(C 2 -C 10 )Alkyl-NH 2 , -(C 2 -C 10 ) alkyl-NH(C 1 -C 3 ) alkyl and -(C 2 -C 10 ) alkyl-N((C 1 -C 3 )Alkyl) 2 and 12. The conjugate of claim 11, wherein o is an integer ranging from 3 to 100.

13. K F The conjugate of claim 12, wherein is H.

14. 14. The conjugate of claim 13, wherein o is in the range of 8 to 30.

15. 15. The conjugate of claim 14, wherein o is in the range of 8 to 16.

16. 16. The conjugate of claim 15, wherein o is 10, 11, 12, 13 or 14.

17. 15. The conjugate of claim 14, wherein o is in the range of 20 to 28.

18. 18. The conjugate of claim 17, wherein o is 22, 23, 24, 25 or 26.

19. The linker L comprises a second spacer unit A, the second spacer unit being a group Z, the group Z having the structure: and where: L p is the parallel connector unit; R S are each independently a second polyalkylene glycol unit; M is independently R s and L p is a bond or moiety that connects s* is an integer ranging from 1 to 4; preferably, s* is 1; and 2. The conjugate of claim 1, wherein the wavy lines indicate points of attachment to -Y- and to another part of the linker, if present, or to the drug moiety (-D).

20. M is each independently -NH-, -O-, -S-, -C(O)-O-, -C(O)-NH-, or -(C 1 -C 10 20. The conjugate of claim 19, wherein said alkylene is selected from the group consisting of:

21. 21. The conjugate of claim 20, wherein each M is -O-.

22. R s but each independently has the structure:

20. The conjugate of claim 19, comprising 1 to 100 subunits having the formula:

23. R s However, each independently, and where: indicates the position of M in the group Z; K s -H, -PO 3 H, -(C 1 -C 10 ) alkyl, -(C 1 -C 10 )Alkyl-SO 3 H, -(C 2 -C 10 )Alkyl-CO 2 H, -(C 2 -C 10 ) alkyl-OH, -(C 2 -C 10 )Alkyl-NH 2 , -(C 2 -C 10 ) alkyl-NH(C 1 -C 3 ) alkyl and -(C 2 -C 10 ) alkyl-N((C 1 -C 3 )Alkyl) 2 and 23. The conjugate of claim 22, wherein p is an integer ranging from 1 to 100.

24. R s is each independently a second polyethylene glycol unit comprising at least one ethylene glycol subunit.

25. Second polyethylene glycol unit R s but each independently has the structure:

25. The conjugate of claim 24, comprising 1 to 100 subunits having the formula:

26. R s However, each independently, and where: indicates the position of M in the group Z; K s -H, -PO 3 H, -(C 1 -C 10 ) alkyl, -(C 1 -C 10 )Alkyl-SO 3 H, -(C 2 -C 10 )Alkyl-CO 2 H, -(C 2 -C 10 ) alkyl-OH, -(C 2 -C 10 )Alkyl-NH 2 , -(C 2 -C 10 ) alkyl-NH(C 1 -C 3 ) alkyl and -(C 2 -C 10 ) alkyl-N((C 1 -C 3 )Alkyl) 2 and 26. The conjugate of claim 25, wherein p is an integer ranging from 1 to 100.

27. 2. The conjugate of claim 1, wherein the linker L is cleavable.

28. 28. The conjugate of claim 27, wherein the linker L is cleavable by a protease, glucuronidase, sulfatase, phosphatase, esterase, or by disulfide reduction.

29. 29. The conjugate of claim 28, wherein the linker L is cleavable by a protease, preferably by a cathepsin such as cathepsin B.

30. 2. The conjugate of claim 1, wherein the linker L comprises a valine-citrulline or valine-alanine moiety.

31. The linker L is 31. The conjugate of claim 30, wherein # indicates the point of attachment to Y and * indicates the point of attachment to the drug moiety.

32. The linker L is and where: R s is a second polyalkylene glycol unit as defined in claim 19; M is as defined in claim 19; and * indicates the connection point to the Y; and 31. The conjugate of claim 30, wherein ## indicates the point of attachment to the drug moiety.

33. 2. The conjugate of claim 1, wherein the drug moiety is selected from the group consisting of maytansinoids, calicheamicins, tubulysins, amatoxins, dolastatins, and auristatins such as monomethylauristatin E (MMAE) or monomethylauristatin F (MMAF), pyrrolobenzodiazepine dimers, indolino-benzodiazepine dimers, emetine, radioisotopes, therapeutic proteins and peptides (or fragments thereof), kinase inhibitors, CDK inhibitors, histone deacetylase (HDAC) inhibitors, MEK inhibitors, KSP inhibitors, and analogs or prodrugs thereof.

34. 34. The conjugate of claim 33, wherein drug moiety D is an auristatin.

35. 35. The conjugate of claim 34, wherein drug moiety D is monomethyl auristatin E (MMAE) or monomethyl auristatin F (MMAF).

36. 35. The conjugate of claim 34, wherein drug moiety D is monomethyl auristatin E (MMAE).

37. the RBM is an antibody; is a double bond; or is a single bond; V, is absent when is a double bond; or V, is a single bond, then H; X is, If is a double bond, R 3 -C; or X is, If is a single bond, and Y is NH; R 1 But the structure: and a first polyethylene glycol unit having the formula: where: indicates the position of O; K F is H; and o is an integer ranging from 8 to 30; R 3 is H; R 4 is H; L has the following structure: where # indicates the point of attachment to Y and * indicates the point of attachment to the drug moiety (D); D is a drug moiety; m is 1; and 2. The conjugate of claim 1, wherein n is an integer ranging from 1 to 10.

38. 38. The conjugate of claim 37, wherein drug moiety D is monomethyl auristatin E (MMAE) or monomethyl auristatin F (MMAF).

39. 39. The conjugate of claim 38, wherein drug moiety D is monomethyl auristatin E.

40. 40. The conjugate of claim 39, wherein o is in the range of 8 to 16.

41. 41. The conjugate of claim 40, wherein o is 10, 11, 12, 13 or 14.

42. 42. The conjugate of claim 41, wherein n is in the range of 2 to 10, preferably n is 4 or 8.

43. 40. The conjugate of claim 39, wherein o is in the range of 20 to 28.

44. 44. The conjugate of claim 43, wherein o is 22, 23, 24, 25 or 26.

45. 45. The conjugate of claim 44, wherein n is in the range of 2 to 10, preferably n is 4 or 8.

46. Compounds having formula (II): or a pharmaceutically acceptable salt or solvate thereof, During the ceremony: is a triple bond; or is a double bond; V is is absent when is a triple bond; or V is If is a double bond, H or (C 1 -C 8 ) alkyl; X is If is a triple bond, R 3 -C: or X is If is a double bond, and Y is NR 5 , S, O, or CR 6 R 7 and R 1 is a first polyalkylene glycol unit R comprising at least three alkylene glycol subunits F and R 3 is H; or an optionally substituted aliphatic residue or an optionally substituted aromatic residue; R 4 is H; or an optionally substituted aliphatic residue or an optionally substituted aromatic residue; R 5 is H; or an optionally substituted aliphatic residue or an optionally substituted aromatic residue; R 6 is H; or an optionally substituted aliphatic residue or an optionally substituted aromatic residue; R 7 is H; or an optionally substituted aliphatic residue or an optionally substituted aromatic residue; L is a linker; D is a drug moiety; and m is an integer ranging from 1 to 10; The compound or a pharmaceutically acceptable salt or solvate thereof.

47. 1. A method for preparing a conjugate of formula (I), the method comprising: Compound of formula (II): or a pharmaceutically acceptable salt or solvate thereof, During the ceremony: is a triple bond; or is a double bond; V is is absent when is a triple bond; or V is If is a double bond, H or (C 1 -C 8 ) alkyl; X is If is a triple bond, R 3 -C: or X is If is a double bond, and Y is NR 5 , S, O, or CR 6 R 7 and R 1 is a first polyalkylene glycol unit R comprising at least three alkylene glycol subunits F and R 3 is H; or an optionally substituted aliphatic residue or an optionally substituted aromatic residue; R 4 is H; or an optionally substituted aliphatic residue or an optionally substituted aromatic residue; R 5 is H; or an optionally substituted aliphatic residue or an optionally substituted aromatic residue; R 6 is H; or an optionally substituted aliphatic residue or an optionally substituted aromatic residue; R 7 is H; or an optionally substituted aliphatic residue or an optionally substituted aromatic residue; L is a linker; D is a drug moiety; and m is an integer ranging from 1 to 10; a compound of formula (II) or a pharmaceutically acceptable salt or solvate thereof; A thiol-containing molecule of formula (III): During the ceremony, RBM is a receptor binding molecule; and n is an integer ranging from 1 to 20; a thiol-containing molecule of formula (III) By reacting Compounds of formula (I): or a pharmaceutically acceptable salt or solvate thereof, During the ceremony: is the compound of formula (II) is a double bond when is a triple bond; or is the compound of formula (II) is a single bond when is a double bond; V is is absent when is a double bond; or V is If is a single bond, H or (C 1 -C 8 ) alkyl; X is If is a double bond, R 3 -C; or X is If is a single bond, and Y is NR 5 , S, O, or CR 6 R 7 and R 1 is a first polyalkylene glycol unit R comprising at least three alkylene glycol subunits F and R 3 is H; or an optionally substituted aliphatic residue or an optionally substituted aromatic residue; R 4 is H; or an optionally substituted aliphatic residue or an optionally substituted aromatic residue; R 5 is H; or an optionally substituted aliphatic residue or an optionally substituted aromatic residue; R 6 is H; or an optionally substituted aliphatic residue or an optionally substituted aromatic residue; R 7 is H; or an optionally substituted aliphatic residue or an optionally substituted aromatic residue; L is a linker; D is a drug moiety; m is an integer ranging from 1 to 10; and n is an integer ranging from 1 to 20; A compound of formula (I) or a pharmaceutically acceptable salt or solvate thereof The step of generating A method comprising:

48. reducing at least one disulfide bridge of the receptor-binding molecule in the presence of a reducing agent to form a thiol group (SH); 48. The method of claim 47, further comprising:

49. 46. ​​A pharmaceutical composition comprising the conjugate of any one of claims 1 to 45.

50. 50. The pharmaceutical composition of claim 49 for use in a method of treating a disease.

51. 51. The pharmaceutical composition of claim 50, wherein the disease is cancer.

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