Nanocomposition comprising modified exatecan and use thereof

Abstract

This disclosure is directed to payload bioactive agents (PBAs) that include modified exatecans. This disclosure is also directed to bioactive compositions and pharmaceutical compositions for treating cancer. The pharmaceutical compositions comprise a polymer, a targeting bioactive agent (TBA), a PBA comprising a modified exatecan that is covalently linked to the TBA directly or indirectly, a linker that can comprise a cleavable linker, and a pharmaceutical suitable carrier. The pharmaceutical compositions can be antibody-drug conjugates (ADCs) for treating cancers, with the potential for treating tumors having negative or low (AgLow) tumor antigens.

Description

NANOCOMPOSITION COMPRISING MODIFIED EXATECAN AND USE THEREOFCROSS REFERENCE TO RELATED APPLICATIONS

[0001] The present disclosure claims priority to U.S. Application No. 63 / 570,208, filed on 26 March 2024, the contents of which is incorporated herein by reference in its entirety.FIELD OF THE DISCLOSURE

[0002] The present disclosure relates to pharmaceutical compositions that can be used for treatment of a disease in patients in need thereof. The pharmaceutical compositions comprise a modified exatecan and antibody drug conjugate nanoaggregates.BACKGROUND

[0003] Antibody-drug conjugates (ADCs) are one of the fastest growing drug classes in oncology therapy. ADCs are composed of monoclonal antibodies (mAbs) linked to cytotoxic drugs and are designed to improve the therapeutic efficacy of antineoplastic agents by targeted delivery to cells that express the specific target molecules, such as target antigens of interest. ADCs have shown promising results in treating cancers, with multiple drugs approved by the US Food and Drug Administration (FDA) and dozens more currently in preclinical and clinical development. Some approved or to be approved drugs include Gemtuzumab ozogamicin (Mylotarg, Pfizer), an anti- CD33 mAb-calicheamicin conjugate; brentuximab vedotin (Adcetris, Seagen / Takeda), a CD30-specific mAb linked to monomethyl auristatin E (MMAE); polatuzumab vedotin (Polivy, Roche); belantamab mafodotin (Blenrep, GlaxoSmithKline); Trastuzumab emtansine (T- DM1 ; Kadcyla, Roche) for any solid tumor, for early- stage and metastatic, HER2+breast cancer; trastuzumab deruxtecan (ENHERTU®, Daiichi Sankyo / AstraZeneca), a second HER2- targeted ADC; sacituzumab govitecan (Trodelvy, Gilead Sciences), an anti- TROP2 mAb conjugated to SN-38, Datopotamab deruxtecan (Dato-DXd), the active metabolite of irinotecan, for previously treated metastatic triple-negative breast cancer; Enfortumab vedotin (Padcev, Seagen / Astellas Pharma), an MMAE conjugated nectin-4 directed mAb for the urothelial cancer market; and the latest cetuximab sarotalocan (Akalux, Rakuten Medical), an epidermal growth factor receptor (EGFR) targeted mAb conjugated to a dye that induces tumor cell death upon photoactivation. Typical mechanisms of targeted delivery by ADC can involve receptor-mediated endocytosis. For example, the anti-tumor activity of the ADC ENHERTU® (fam-trastuzumab-deruxtecan-nxki) is mainly achieved bythe binding of the human epidermal receptor 2 (HER2) by monoclonal antibody trastuzumab, and delivery of cytotoxic drug deruxtecan via antibody-dependent receptor- mediated endocytosis (RME). The anti-tumor activity of the ADCs requires the tumor cells having specific tumor antigens that can be specifically targeted by the antibody, such as the human epidermal receptor 2 (HER2) in HER2-positive breast cancer. However, the design of clinically effective ADCs has presented challenges for the industry: since the first ADC, Mylotarg® (gemtuzumab ozogamicin), was approved in 2000 by the FDA, there have been only 14 ADCs received market approval so far worldwide after over 30 years of research and development. Over 100 ADC candidates are still being investigated in clinical stages at present.

[0004] There is a continued need for new drugs and methods to deliver drugs more effectively or to improve upon tissue specificity and cytotoxicity of drugs.SUMMARY

[0005] In some cases, present disclosure is directed to a payload bioactive agent (PBA) comprising a formula selected from:a pharmaceutically acceptable salt thereof, a solvate thereof, a prodrug thereof, an isomer thereof, an isotope derivative thereof, and a combination thereof; wherein, v and w each is an integer, v=1-100; w=1-1000; R, R’, R”, R3, R4, R3’, R4’, Rs, Re, R7, Rs, and R15 each is independently H, D, C1-C100 saturated or unsaturated hydrocarbon, C1-C100 saturated or unsaturated hydrocarbon having at least one D, C1- C100 saturated or unsaturated hydrocarbon having at least one heteroatom, C1 -C100 saturated or unsaturated hydrocarbon having at least one heteroatom and at least one D, aromatic group, substituted aromatic group, or a combination thereof; R9 is selected from H, D, C1-C100 saturated or unsaturated hydrocarbon, C1-C100 saturated or unsaturated hydrocarbon having at least one D, C1-C100 saturated or unsaturated hydrocarbon having at least one heteroatom, C1-C100 saturated or unsaturated hydrocarbon having at least one heteroatom and at least one D, OH, NH2, Cl, F, I, and Br; R10 is selected from H, Cl, F, I, Br, and methoxy; Rn is OH, OR12, SH, SR13, NH2, NHD, ND2, and NRuR’u, wherein R12 is an alcohol protecting group, R13 is a thiol protecting group, and Ru and R’14 each is independently H, D, or an amine protecting group; the heteroatom is selected from N, O, S, P, Cl, Br, I, and a combination thereof; and X is O, S, or NH.

[0006] This disclosure is also directed to a pharmaceutical composition comprising the PBA. In some cases, this disclosure is further directed to a pharmaceutical composition comprising the PBA, a targeting bioactive agent (TBA) comprising binding affinity to a target molecule or a target cell; and wherein the PBA is reacted and covalently linked to the TBA directly or indirectly.

[0007] In some cases, the pharmaceutical composition can further comprise a polymer, a linker, optionally a coupler, or a combination thereof, wherein the PBA is covalently linked to the polymer, the linker, or optionally, the coupler when present, or a combination thereof, wherein the polymer comprises a peptide, a polyethylene glycol (PEG), polyoxazoline (POX), or a combination thereof, wherein the POX comprises a linear portion, a branched portion, or a combination thereof, and wherein the POX comprisespoly(2-methyloxazoline), poly(2-ethyloxazoline), poly(2-propyloxazoline), poly(2- isopropyloxazoline), poly(2-substituted oxazoline), or a combination thereof.

[0008] In some cases, this disclosure is further directed to an antibody drug conjugate (ADC).

[0009] In some cases, the present disclosure is directed to a method for treating a disease of a subject in need thereof comprising administering the subject with an effective dose of a pharmaceutical composition disclosed herein.

[0010] In some cases, the present disclosure is directed to a use of the pharmaceutical composition, the bioactive composition or a Nano-ADC disclosed herein for manufacturing of a medicament for treating or preventing a disease.BRIEF DESCRIPTION OF THE FIGURES

[0011] FIG. 1A-FIG. 1 D. Schematic illustrations of representative examples of the pharmaceutical composition having nanoaggregates comprising a targeting bioactive agent (TBA), a payload bioactive agent (PBA) and a polymer. FIG. 1A: An example of pharmaceutical composition and nanoaggregates that comprise no cleavable linker between the TBA and PBA. FIG. 1 B: An example of the pharmaceutical composition and nanoaggregates that comprise at least one cleavable linker between the TBA and PBA. FIG. 1C: An example of the pharmaceutical composition and nanoaggregates that comprise a polymer, at least one cleavable linker between the TBA and PBA. FIG. 1D: Schematic illustration of a representative example of nanoparticle complex that can comprise multiple nanoparticles.

[0012] FIG. 2A-FIG. 2E. Schematic illustrations of representative examples of a pharmaceutical composition having the polymers covalently linked to the TBA and PBA, where the (R1 ) and (R2) represent the reacted first terminal group and the reacted second terminal group of the polymer, respectively; open rectangle: Linker L1 ; hatched rectangle: Linker L2; solid rectangle: the polymer of this disclosure; crossed circle: cleavable bond; solid circle: PBA; and each TBA can be covalently linked to one or more PBAs, n is an integer in a range of from 1 to 100. FIG. 2A: An example of a pharmaceutical composition comprising no cleavable linker. FIG. 2B: An example of the pharmaceutical composition comprising at least one cleavable linker in the Linker L1 between the TBA and the polymer. FIG. 2C: An example of the pharmaceutical composition comprising at least one cleavable linker in the Linker L2 between the PBA and the polymer. FIG. 2D: An example of the pharmaceutical composition comprising at least one cleavable linker in the Linker L1 between the TBA and the polymer and at least one cleavable linker in the Linker L2between the PBA and the polymer. FIG. 2E: A schematic illustration of a representative example of a pharmaceutical composition having multiple different polymers and PBAs.

[0013] FIG. 3A-FIG. 3D. Schematic illustrations of representative examples of the pharmaceutical composition comprising the polymers having asymmetric branches covalently linked to one TBA and multiple PBAs. FIG. 3A: An example of the pharmaceutical composition comprising no cleavable linker. FIG. 3B: An example of the pharmaceutical composition comprising at least one cleavable linker in the Linker L1 between the TBA and the polymer. FIG. 3C: An example of the pharmaceutical composition comprising more than one cleavable linkers in the Linker L2 between the PBA and the polymer. FIG. 3D: An example of the pharmaceutical composition comprising at least one cleavable linker in the Linker L1 between the TBA and the polymer and more than one cleavable linkers in the Linker L2 between the PBA and the polymer.

[0014] FIG. 4A-FIG. 4D. Schematic illustrations of representative examples of the pharmaceutical composition comprising the polymers having symmetric branches covalently linked to one TBA and multiple PBAs. FIG. 4A: An example of the pharmaceutical composition comprising no cleavable linker. FIG. 4B: An example of the pharmaceutical composition comprising at least one cleavable linker in the Linker L1 between the TBA and the polymer. FIG. 4C: An example of the pharmaceutical composition comprising more than one cleavable linkers in the Linker L2 between the PBA and the polymer. FIG. 4D: An example of the pharmaceutical composition comprising at least one cleavable linker in the Linker L1 between the TBA and the polymer and more than one cleavable linkers in the Linker L2 between the PBA and the polymer.

[0015] FIG. 5. Schematic illustration of representative examples of the formation of polymer-PBA aggregates. The pharmaceutical composition can comprise at least a cleavable linker in Linker L1 .

[0016] FIG. 6A-FIG. 6C. Schematic illustration of representative examples of the formation of nanoaggregates after cleavage. FIG. 6A: The Nano-ADC can comprise cleavable linkers in both linkers L1 and L2 forming polymer nanoaggregates after cleavage. FIG. 6B: The Nano-ADC can comprise modified exatecan forming nanoaggregates after cleavage. FIG. 6C: The Nano-ADC can comprise modified exatecan forming nanoaggregates including the polymer after cleavage.

[0017] FIG. 7A-FIG. 7H. Schematic representations of examples of antibody-drug conjugates (ADCs). FIG. 7A: A schematic example of an ADC having Ab-Polymer-Linker- PBA structure. FIG. 7B: A schematic representation of an example of an ADC having Ab- Linker-Polymer-PBA structure. FIG. 7C: A schematic representation of an example of anADC having Ab-PEG-GGFG-mExa structure. FIG. 7D: A schematic representation of an example of an ADC having Ab-GGFG-PEG-mExa structure. FIG. 7E: A schematic representation of an example of an ADC having Ab-PEG-VCPAB-mExa structure. FIG. 7F: A schematic representation of an example of an ADC having Ab-VCPAB-PEG-mExa structure. FIG. 7G: A schematic representation of an example of an ADC having Ab-PEG- VAPAB-mExa structure. FIG. 7H: A schematic representation of an example of an ADC having Ab-VAPAB-PEG-mExa structure.

[0018] FIG. 8A-FIG. 8L. Schematic illustrations of representative examples of the pharmaceutical composition comprising at least a branched or linear polymer side chain that has one or more free second terminal groups that each is free from PBAs, where R1 and R2 represent the first terminal group and the second terminal group, respectively. FIG. 8A: An example of a pharmaceutical composition comprising no cleavable linker between the TBA, polymer and PBA with a branched polymer side chain attached to the polymer. FIG. 8B: An example of a pharmaceutical composition comprising no cleavable linker between the TBA, PBA with a branched polymer side chain attached to the TBA. FIG. 8C: A schematic illustration of a representative example of a pharmaceutical comprising at least one cleavable linker and a branched polymer side chain attached to the polymer. FIG. 8D: A schematic illustration of a representative example of a pharmaceutical comprising at least one cleavable linker and a branched polymer side chain attached to the TBA. FIG. 8E: A schematic illustration of a representative example of a pharmaceutical comprising no cleavable linker and a linear polymer side chain attached to the polymer. FIG. 8F: A schematic illustration of a representative example of a pharmaceutical comprising at least one cleavable linker and a branched polymer side chain attached to the TBA. FIG. 8G: A schematic illustration of a representative example of a pharmaceutical comprising at least one cleavable linker and a linear polymer side chain attached to the polymer. FIG. 8H: A schematic illustration of a representative example of a pharmaceutical comprising at least one cleavable linker and a linear polymer side chain attached to the TBA. FIG. 81: A schematic illustration of a representative example of a pharmaceutical comprising at least one cleavable linker and a linear or branched polymer side chain attached to the linker, such as Linker L1 that is cleavable. FIG. 8J: A schematic illustration of a representative example of a pharmaceutical comprising at least one cleavable linker and a linear or branched polymer side chain attached to the linker, such as Linker L2 that is cleavable. FIG. 8K: A schematic illustration of a representative example of a pharmaceutical comprising at least one cleavable linker and a linear or branched polymer side chain attached to the linker, such as Linker L1 thatis not cleavable. FIG. 8L: A schematic illustration of a representative example of a pharmaceutical comprising at least one cleavable linker and a linear or branched polymer side chain attached to the linker, such as Linker L2 that is not cleavable.

[0019] FIG. 9A-FIG. 9L. Schematic illustrations of representative examples of the pharmaceutical composition comprising at least a branched or linear polymer side chain that has one or more PBAs attached at the second terminal groups, where R1 and R2 represent the first terminal group and the second terminal group, respectively, that each is free from a PBA or reacted to a PBA (for simplicity, not all R2 groups are shown). The one or more first PBAs (3) and the one or more second PBAs (3’) can be the same or different. FIG. 9A: An example of a pharmaceutical composition comprising no cleavable linker between the TBA, polymer, and PBA with a branched polymer side chain attached to the polymer. FIG. 9B: An example of a pharmaceutical composition comprising no cleavable linker between the TBA, polymer, and PBA with a branched polymer side chain attached to the TBA. FIG. 9C: A schematic illustration of a representative example of a pharmaceutical comprising at least one cleavable linker and a branched polymer side chain attached to the polymer. FIG. 9D: A schematic illustration of a representative example of a pharmaceutical comprising at least one cleavable linker and a branched polymer side chain attached to the TBA. FIG. 9E: A schematic illustration of a representative example of a pharmaceutical comprising no cleavable linker and a linear polymer side chain attached to the polymer. FIG. 9F: A schematic illustration of a representative example of a pharmaceutical comprising at least one cleavable linker and a branched polymer side chain attached to the TBA. FIG. 9G: A schematic illustration of a representative example of a pharmaceutical comprising at least one cleavable linker and a linear polymer side chain attached to the polymer. FIG. 9H: A schematic illustration of a representative example of a pharmaceutical comprising at least one cleavable linker and a linear polymer side chain attached to the TBA. FIG. 9I: A schematic illustration of a representative example of a pharmaceutical comprising at least one cleavable linker and a linear or branched polymer side chain attached to at least one PBA and the linker, such as Linker L1 that is cleavable. FIG. 9J: A schematic illustration of a representative example of a pharmaceutical comprising at least one cleavable linker and a linear or branched polymer side chain attached to at least a PBA and the linker, such as Linker L2 that is cleavable. FIG. 9K: A schematic illustration of a representative example of a pharmaceutical comprising at least one cleavable linker and a linear or branched polymer side chain attached to at least a PBA and the linker, such as Linker L1 that is not cleavable. FIG. 9L: A schematic illustration of a representative example of apharmaceutical comprising at least one cleavable linker and a linear or branched polymer side chain attached to at least a PBA and the linker, such as Linker L2 that is not cleavable.

[0020] FIG. 10A-FIG. 10K. Representative examples of Nano-antibody-drug conjugate (Nano-ADC) comprising polymer, cleavable linker and modified exatecan. FIG. 10A: Nano-ADC Formula ID 3001. FIG. 10B: Nano-ADC Formula ID 3002. FIG. 10C: Nano- ADC Formula ID 3003. FIG. 10D: Nano-ADC Formula ID3004. FIG. 10E: Nano-ADC Formula ID 3005. FIG. 10F: Nano-ADC Formula ID 3006. FIG. 10G: Nano-ADC Formula ID 3007. FIG. 10H: Nano-ADC Formula ID 3008. FIG. 101: Nano-ADC Formula ID 3009. FIG. 10J: Nano-ADC Formula ID 3010. FIG. 10K: Nano-ADC Formula 3011.

[0021] FIG. 11A-FIG. 11F. Representative examples of Nano-ADCs having GGFG cleavable linkers and at least a modified exatecan. FIG. 11 A: A Nano-ADC having Formula ID 3012. FIG. 11B: A Nano-ADC having Formula ID 3013. FIG. 11C: A Nano- ADC having Formula ID 3014. FIG. 11D: A Nano-ADC having Formula ID 3015. FIG. 11E: A Nano-ADC having Formula ID 3016. FIG. 11F: A Nano-ADC having Formula ID 3017. When applicable, m is an integer in a range of from 1 - 100, and n is an integer in a range of from 1 to 100. R’ is as disclosed herein.

[0022] FIG. 12A-FIG. 12U. Representative examples of Nano-ADCs having VAPAB cleavable linkers and a modified exatecan. FIG. 12A: A Nano-ADC having Formula ID 3018. FIG. 12B: A Nano-ADC having Formula ID 3019. FIG. 12C: A Nano-ADC having Formula ID 3020. FIG. 12D: A Nano-ADC having Formula ID 3021. FIG. 12E: A Nano- ADC having Formula ID 3022. FIG. 12F: A Nano-ADC having Formula ID 3023. FIG. 12G: A Nano-ADC having Formula ID 3024. FIG. 12H: A Nano-ADC having Formula ID 3025. FIG. 121: A Nano-ADC having Formula ID 3026. FIG. 12J: A Nano-ADC having Formula ID 3027. FIG. 12K: A Nano-ADC having Formula ID 3028. FIG. 12L: A Nano-ADC having Formula ID 3029. FIG. 12M: A nano-ADC having Formula ID 3030. FIG. 12N: A Nano- ADC having Formula ID 3031 . FIG. 120: A Nano-ADC having Formula ID 3032. FIG. 12P: A Nano-ADC having Formula ID 3033 that comprises an antibody to B7H3. FIG. 12Q: A Nano-ADC having Formula ID 3034. FIG. 12R: A Nano-ADC having Formula ID 3035. FIG. 12S: A Nano-ADC having Formula ID 3036. FIG. 12T: A Nano-ADC having Formula ID 3037. FIG. 12U: A Nano-ADC having Formula ID 3038. When applicable, m is an integer in a range of from 1 - 100, y is an integer in a range of from 1 to 100, and n and n’ each is independently an integer in a range of from 1 to 100. R and R’ each is as disclosed herein.

[0023] FIG. 13A-FIG. 13C. Schematic illustrations of tumor cell cytotoxicity mechanism model. FIG. 13A: Traditional receptor-mediated endocytosis (RME). FIG. 13B: A representative example of Nanoparticle-mediated cytotoxicity (NMC) to deliver AgHightumor cytotoxicity to tumor cells having a high level of a tumor antigen (AgHightumor cells) and AgLowtumor cytotoxicity to tumor cells free from any tumor antigen (AgLowtumor cells). FIG. 13C: A representative example of Nanoparticle-mediated immune modification (NMIM) to enhance immune system response to delivery AgHightumor cytotoxicity to AgHightumor cells and AgLowtumor cytotoxicity to AgLowtumor cells.

[0024] FIG. 14A-FIG. 14B. Nano-ADC DAR measurements. FIG. 14A: A representative product of Nano-ADC Formula ID 3024K with a measured DAR at 8.7. FIG. 14B: A representative product of Nano-ADC Formula ID 3024K with a measured DAR at 9.0.

[0025] FIG. 15. Cytotoxicity of modified exatecans in vitro. Cell viability assay data of unmodified Exatecan (solid circle), deruxtecan (solid square), TFE (Formula ID 1005) (solid diamond), PrE (Formula ID 1004) (overlaid square and solid circle), BzE (Formula ID 1012) (open triangle), cHME (Formula ID 1013) (small solid dot), nBE (Formula ID 1014) (+), iBE (Formula ID 1015) (x), nPE (Formula ID 1016) (open circle), bPE (Formula ID 1011 ) (overlaid open square and asterisk).

[0026] FIG. 16A-FIG. 16B. Cytotoxicity measured with cell viability assay. FIG. 16A: Assays with SKBR-3 cells. FIG. 16B: Assays with NCI-N87 cells.

[0027] FIG. 17A-FIG. 17B. Tumor size measurements in SCID CDX mouse models having low HER2 levels. FIG. 17A: SCID CDX mouse with HCT116 tumor cells and Formula ID 3027 and 3029. FIG. 17B: SCID CDX mouse with HT29 cells and Formula ID 3080 (human IgG, herein HlgG,” as a control), Enhertu and Formula ID 3036.

[0028] FIG. 18A-FIG. 18B. Tumor size measurements in SCID CDX mouse models with tumor cells having high HER2 levels. FIG. 18A: SCID CDX mouse with NCI-N87 tumor cells having high HER2 levels and Formula ID 3024K and 3027. FIG. 18B: SCID CDX mouse with NCI-N87 tumor cells having high HER2 levels and Formula ID 3035.

[0029] FIG. 19A-FIG. 19B. Tumor size measurements in SCID CDX mouse models with tumor cells having medium or high B7H3 levels. FIG. 19A: SCID CDX mouse with NCI- H446 tumor cells having medium B7H3 expression level and Nano-ADC Formula ID 3033. FIG. 19B: SCID CDX mouse with NCI-H526 tumor cells having high B7H3 expression level and Nano-ADC Formula ID 3033.

[0030] FIG. 20. Cytotoxicity of the Nano-ADCs measured via multiple injections using SCID mouse models xenografted with the HCT 116 tumor cells derived from colon cancer that are considered to have low HER2 expression levels (HER2Low).

[0031] FIG. 21. Tissue distribution of the PBA in SCID CDX mouse models induced with NCI-N87 tumor cells. Enhertu: an ADC fam-trastuzumab deruxtecan (ENHERTU®). Formula ID 3027: Nano-ADC.DETAILED DESCRIPTION

[0032] Features and advantages of the present disclosure will be more readily understood, by those of ordinary skill in the art, from reading the following detailed description. It is to be appreciated that certain features of the disclosure, which are described above and below in the context of separate embodiments, may also be provided in combination in a single embodiment. Conversely, various features of the disclosure that are, for brevity, described in the context of a single embodiment, may also be provided separately or in any combination or sub-combination. In addition, references in the singular may also include the plural (for example, “a” and “an” may refer to one, or one or more) unless the context specifically states otherwise.

[0033] Use of numerical values in the various ranges specified in this application, unless expressly indicated otherwise, are stated as approximations as though minimum and maximum values within the stated ranges were both proceeded by the word, "about.” In this manner, slight variations above and below the stated ranges can be used to achieve substantially the same results as values within the ranges. Also, disclosure of ranges is intended as a continuous range including every value between the minimum and maximum values and including the minimum and maximum cited values.

[0034] The drug solubility in the instant disclosure is defined as, relative to parts of solvent required to solubilize one part of bioactive agent or drug, <30 (soluble), 30-100 (poorly soluble) and >100 (insoluble). Water solubility is defined herein as, relative to parts of water required to solubilize one part of bioactive agent or drug, <30 (water soluble), 30- 100 (poorly water soluble) and >100 (water insoluble).

[0035] The term “polymer” refers to any polymer suitable for this disclosure as defined above and hereafter. In examples, a polymer can comprise a polypeptide, polyethylene glycol (PEG), polyoxazoline or modified polyoxazoline as disclosed herein. In further examples, the polymer can comprise a modified polyoxazoline, which can comprise one or more second terminal groups, such as an -NH2, -NH, -NHs+, other basic groups or a combination thereof. In some cases, polymer can comprise a single amino acid residue, dipeptide, tripeptide, or polypeptide. In some cases, polymer can comprise a polypeptide that comprises a lysine (Lys or K), glutamic acid (Glu or E), glutamine (Gin or Q), arginine (Arg or R), valine (Vai or V), or a combination thereof. In some cases. The polymer cancomprise lysine-valine (K-V), glutamic acid-valine (E-V), glutamic acid-arginine (E-R), valine-citrulline (VC), valine-alanine (VA), or a combination thereof. Peptides disclosed herein can comprise amino acid derivatives, modified amino acids, modified amino acid residues, or a polypeptide comprising in a range of 1-100 or more units of same or different amino acids, same or different amino acid residues, same or different amino acid derivatives, same or different modified amino acids, same or different modified amino acid residues, including naturally occurring nonessential amino acid such as L-citrulline, or a combination thereof. In some cases, a polymer can comprise a lysine, glutamic acid, glutamine, arginine, valine, or a combination thereof. The polymer can comprise a linear polymer, a branched polymer, a symmetrically branched polymer, an asymmetrically branched polymer, a dendrimer, a dendrigraft polymer, a comb-branched polymer, a star- branched polymer, or a combination thereof. The polymer is water soluble. In examples, the polymer can be dissolved in water to produce a 12% weight percent or higher water solution. In some cases, in a range of from 0.01 % to 100% of the second terminal group is free from primary amine. In some cases, in a range of from 0.01 % to 100%, 0.1 % to 100%, or 1 % to 100%, of the second terminal group is free from primary amine. In some cases, in a range of from 1 % to 100% of the second terminal group can comprise hydroxyl group. All percentages are based on the total number of the second terminal groups.

[0036] The term “bioactive agent” or “bioactive agents” refers to a molecule, a compound, a complex of one or more compounds or molecules, or a combination thereof that can provide a biological activity in vivo, in vitro, or a combination thereof. A pharmaceutical composition can comprise one or more bioactive agent such as pharmaceutically active agents (PAAs) or active pharmaceutical ingredients (APIs), and other bioactive or inert compounds that can include emollients, bleaching agents, antiperspirants, pharmaceuticals, moisturizers, scents, colorants, pigments, dyes, antioxidants, oils, fatty acids, lipids, inorganic salts, organic molecules, opacifiers, vitamins, pharmaceuticals, keratolytic agents, UV blocking agents, tanning accelerators, depigmenting agents, deodorants, perfumes, insect repellants, or a combination thereof. Some examples of bioactive agents are described in detail in this disclosure.

[0037] The term “targeting bioactive agent” or “TBA” refers to a compound or a molecule that can comprise at least a targeting moiety having binding affinity to a target molecule, a target cell or a target molecule of a target cell. A target molecule can be on the surface of a target cell, in the cytosol of a target cell, in the nucleus of a target cell, in one or more organelles of a target cell, on the membranes of one or more organelles of a target cell, or a combination thereof. As used herein, the term “organelle” refers to a small structurein a cell that is surrounded by a membrane and can have a specific function. In some cases, unlimiting examples of organelles can include the nucleus (a structure that contains the cell’s DNAs and RNAs), mitochondria (structures that make energy for the cell), golgi complex of a cell, and lysosomes (sac-like containers filled with enzymes that digest and help recycle molecules in the cell). In some cases, a TBA can comprise peptide, a protein, DNA, RNA, nucleic acids, oligo nucleic acids, mRNA, siRNA, sgRNA, an antibody selected from a monoclonal antibody, a fragment of a monoclonal antibody, a polyclonal antibody, a fragment of a polyclonal antibody, a synthetic antibody, a fragment of a synthetic antibody, antigen-binding portions thereof, or a combination thereof, a ligand of cell surface receptor, an agonist, an activator, an inhibitor, an antagonist, an antisense nucleic acid, a naked nucleic acid, a nucleic acid, a ribozyme, a virus, a virus-like particles, T-cell receptor (TCR), chimeric antigen receptor or chimeric antigen T-cell receptor (CAR-T), cytokines, interferon a, interferon a2a, interleukins, granulocyte colony stimulating factor (G-CSF), neupogen (Filgrastim), or a combination thereof. In some cases, an antibody can comprise alemtuzumab, anti-CD19 antibodies, anti-CD20 antibodies, anti-CTLA-4 (cytotoxic T lymphocyte-associated antigen) antibodies, anti-LAG3 (lymphocyte activation gene-3) antibodies, anti-PD1 antibodies KEYTRUDA® (pembrolizumab), anti-PD-L1 antibodies, anti-TIM-3 (T cell immunoglobulin and mucin domain-3) antibodies, atezolizumab, BAVENCIO® (avelumab), bevacizumab, blinatumomab, brentuximab, cetuximab, daratumumab, datopotamab, denosumab, dinutuximab, disitamab, durvalumab, elotuzumab, enfortumab, gemtuzumab, ibritumomab, IMFINZI® (durvalumab), inotuzumab, ipilimumab, lectins, loncastuximab, mirvetuximab, necitumumab, obinutuzumab, ofatumumab, olaratumab, OPDIVO® (nivolumab), panitumumab, pembrolizumab, pertuzumab (anti-HER2), polatuzumab, ramucirumab, rovalpituzumab, rituximab, rituximab, sacituzumab, siltuximab, toripalimab, tislelizumab (BGB-A317), tisotumab, TECENTRIQ® (atezolizumab), tositumomab, trastuzumab (anti-HER2), one or more antibodies against target molecules comprising 5T4, ALPPL2, AXL, B7H3, B7H4, B7H6, B7S1 , B7x, BCAM, BCMA, CCL2, CCL5, CD146, CD25, CD27 ligand, CD276, CD47, CD70, CDH3, CDH6, CEACAM5, CEACAM6, CLDN1 , CLDN18.2, CLDN6, DLK1 , DLL3, EGFR, EGFRVIII, EpCAM, ErbB1 , ErbB2, ErbB3, FAT2, FOLR1 , GPC3, GPR87, GRM8, GRP56, HER2, HER3, HGFR, ITGB6, LIV-1 , Ly6G6D, LY6G6F, LYPD3, MCAM, MET, MSLN, MUC16, MUC18, PSMA, ROR1 , SEZ6, SLC39A6, SLC7A11 , SLCO1 B3, STEAP1 , Tacstd2, TLR4, TMPRSS4, TROP2, UFO, VEGF, VTCN1 , ZIP6, a bispecific antibody thereof, a tri-specific antibody thereof, a tetra-specific antibody thereof, a fragment thereof, an antigen-binding portionthereof, or a combination thereof. Antibodies can be bispecific, tri-specific and tetraspecific antibodies, or a combination thereof and can include a monoclonal antibody, a fragment of a monoclonal antibody, a polyclonal antibody, a fragment of a polyclonal antibody, a synthetic antibody, a fragment of a synthetic antibody, antigen-binding portions thereof, or a combination thereof. The term “a bispecific antibody thereof”, “a tri- specific antibody thereof”, “a tetra-specific antibody thereof”, “a fragment thereof” or “an antigen-binding portion thereof” refers to a bispecific antibody of any one of the antibodies disclosed herein, a tri-specific antibody of any one of the antibodies disclosed herein, and a tetra-specific antibody of any one of the antibodies disclosed herein, a fragment of any one of the antibodies disclosed herein, and an antigen-binding portion of any one of the antibodies disclosed herein.

[0038] The term “payload bioactive agent” or “PBA” refers to a natural or synthetic small molecule-based drug, inorganic-based drug, a toxin molecule having cytotoxicity, a molecule having cytotoxicity to a target cell, a derivative thereof, or a combination thereof. In some cases, a PBA can be selected from 5-FU (Fluorouracil), alkylators, antimetabolites, cabazitaxel, calicheamicin, calicheamicin derivatives, camptothecin, camptothecin derivatives, capecitabine, cell division inhibitors or microtubule inhibitors, ci profl oxaxi n, cyclophosphamide, deruxtecan, deruxtecan derivatives, DNA cleavage agent, DNA crosslinking agent, DNA replicating inhibitor, docetaxel, dolastatin analogs, doxorubicin, doxorubicin hydrochloride, duocarmycin analogs, etoposide, everolimus, exatecan (in certain formulas throughout this disclosure, exatecan is also referred to as “E” as an abbreviation), exatecan derivatives, topoisomerase inhibitors, gemcitabine, gemcitabine hydrochloride, irinotecan (CPT-11 ), irinotecan hydrochloride, larotaxel, maytansinoids (DM1 , DM4), DM1 (antimicrotubule agent T-DM1 , a derivative of maytansine), DM4 (Ravtansine), milataxel, MMAE (monomethyl auristatin E), MMAF (monomethyl auristatin F), MMAH (monomethyl auristatin H), mTOR inhibitor, one or more ATP-competitive mTORC1 / 2 inhibitors, one or more dual PI3K-mTOR inhibitors, ortataxel, ozogamicin, pemetrexed, pyrrolobenzodiazepine dimer (PBD), SG3199 (a cytotoxic DNA minor groove interstrand crosslinking pyrrolobenzodiazepine (PBD) dimer), rapamycin, ridaforolimus, SN-38, taxane, tesetaxel, Topoisomerase 1 (Top 1) inhibitor, Topoisomerase 2 (Top 2) inhibitor, topotecan, torin-1 , torin-2, vinblastine, vinca alkaloids, vincristine, vinorelbine, vistusertib, zotarolimus, biological toxins, ricin, abemaciclib, abiraterone acetate, acalabrutinib, afatinib dimaleate, aldesleukin (IL-2 or interleukin-2), alectinib hydrochloride, altretamine, amifostine, aminolevulinic acid hydrochloride, anastrozole, apremilast, aprepitant, arsenic trioxide, asparaginase erwiniachrysanthemi, axicabtagene ciloleucel, axitinib, azacitidine, belinostat, bendamustine hydrochloride, bexarotene, bicalutamide, bleomycin sulfate, bortezomib, bosutinib monohydrate, brigatinib, busulfan, cabozantinib s-malate, carboplatin, carfilzomib, carmustine, carmustine implant, ceritinib, chlorambucil, cisplatin, cladribine, clofarabine, cobimetinib fumarate, copanlisib dihydrochloride, crizotinib, cytarabine, cytarabine liposome, cytarabine, daunorubicin, dabrafenib mesylate, dacarbazine, dactinomycin, dasatinib, daunorubicin citrate, daunorubicin hydrochloride, decitabine, defibrotide sodium, degarelix acetate, denileukin diftitox, dexamethasone, edoxaban tosylate, eltrombopag olamine, enasidenib mesylate, enzalutamide, epirubicin hydrochloride, eribulin mesylate, erlotinib hydrochloride, estramustine phosphate sodium, exemestane, Dxd (deruxtecan, a derivative of exatecan), filgrastim, floxuridine, fludarabine phosphate, fluorouracil, flutamide, fulvestrant, gefitinib, glucarpidase, goserelin acetate, hydroxyurea, ibrutinib, idarubicin hydrochloride, idelalisib, ifosfamide, mesna, imatinib mesylate, imiquimod, ixabepilone, ixazomib citrate, lanreotide acetate, lapatinib ditosylate, lenalidomide, lenvatinib mesylate, letrozole, leucovorin calcium, leuprolide acetate, lomustine, mechlorethamine hydrochloride, megestrol acetate, melphalan, mercaptopurine, mesna, methotrexate, methotrexate sodium, methylnaltrexone bromide, midostaurin, mitomycin, mitotane, mitoxantrone hydrochloride, naldemedine tosylate, nelarabine, neratinib maleate, netupitant, palonosetron hydrochloride, nilotinib hydrochloride monohydrate, nilutamide, niraparib tosylate, olaparib, omacetaxine mepesuccinate, ondansetron, ondansetron hydrochloride, osimertinib mesylate, oxaliplatin, paclitaxel, palbociclib, palifermin, palonosetron hydrochloride, pamidronate disodium, panobinostat lactate, pazopanib hydrochloride, pegademase bovine, pegaspargase, pegfilgrastim, peginterferon alfa-2b, pemetrexed disodium, pentostatin, plerixafor, plicamycin, pomalidomide, ponatinib hydrochloride, pralatrexate, prednisone, procarbazine hydrochloride, propranolol hydrochloride, radium 223 dichloride, raloxifene hydrochloride, rasburicase, regorafenib, rolapitant hydrochloride, romidepsin, romiplostim, rucaparib camsylate, ruxolitinib phosphate, sipuleucel-t, sonidegib phosphate, sorafenib tosylate, streptozocin, sunitinib malate, talc, talimogene laherparepvec, tamoxifen citrate, temozolomide, temsirolimus, teniposide, thalidomide, thioguanine, thiotepa, tisagenlecleucel, topotecan hydrochloride, toremifene citrate, trabectedin, trametinib dimethyl sulfoxide, uridine triacetate, valrubicin, vandetanib, vemurafenib, venetoclax, vinblastine sulfate, vincristine sulfate, vinorelbine tartrate, vismodegib, vorinostat, ziv-aflibercept, zoledronic acid, or a combination thereof. In somecases, a cytotoxic compound or molecule can be preferred. In some cases, a toxin, such as ricin, can be suitable.

[0039] The term “ADC” refers to antibody-drug conjugate that is typically composed of a monoclonal antibody (mAbs) covalently attached to a cytotoxic drug via a chemical linker. In some examples, the ADC can comprise an anti-HER2 monoclonal antibody. The terms “HER2,” “HER2,” “ErbB2,” or “Neu” refer to a transmembrane glycoprotein in the ErbB family of tyrosine kinase receptors for EGF superfamily growth factors. ErbB2 is widely expressed in epithelial cells and over-expressed in a large number of breast carcinomas and is considered as a tumor antigen (Ag) present in tumor cells. ErbB2 plays roles in development, cancer, communication at the neuromuscular junction, and regulation of cell growth and differentiation. The ErbB2 / ErbB3 heterodimer is expressed in the majority of breast, skin, ovary and gastrointestinal tumors and transduces a highly mitogenic signal in response to neuregulin 1 (NRG1 ; heuregulin 1 ) or NRG2. ErbB3, ErbB2 and neuregulin are all required for formation of the sympathetic nervous system. ADCs can potentially, via specificity of a monoclonal antibody, selectively deliver highly cytotoxic small molecules to targeted cancer cells, leading to an enhanced therapeutic index through increased antitumor activity (tumor cytotoxicity) particularly the AgHightumor cytotoxicity to tumor cells having high levels of tumor antigen (AgHightumor cells) and decreased off- target toxicity, i.e., cytotoxicity to non-tumor cells (non-tumor cytotoxicity). Anti-HER2 ADCs trastuzumab emtansine and trastuzumab deruxtecan are examples of the ADCs for the treatment of patients with human epidermal growth factor receptor 2 (HER2)- positive breast cancer. Currently, a total of 29 ADC candidates are being investigated in different stages of clinical development for the treatment of HER2-positive breast cancer. Currently, no anti-HER2-ADCs have been reported for the treatment of HER2-negative breast cancer or other HER2-negative / HER2 low cancers. ADC can also comprise antibodies against target molecules comprising HER3, BCMA, PSMA, EpCAM, EGFRVIII, ROR1 , MSLN, TROP2, B7H4, CLDN18.2, 5T4, CEACAM5, CEACAM6, CEACAM5 / 6, CD25, DLK1 , B7H3, MUC16, FOLR1 , DLL3, LY6G6D, CCL2, CCL5, EGFR, CD47, VEGF, or a combination thereof. Antibodies can be bispecific, tri-specific and tetraspecific antibodies, or a combination thereof. ADCs disclosed in references are referred to as ’’traditional ADCs” in this disclosure.

[0040] The term “AgHightumor cells” refers to cells or tumors that have abnormally high levels of the tumor antigen (Ag). For example, a HER2+tumor cells can have abnormally high level of the tumor antigen HER2 protein. About 14% of new female breast cancer cases in the United States between 2015 and 2019 are HER2+. Treatment for HER2+cancers may include the targeted therapy with an ADC fam-trastuzumab deruxtecan (ENHERTU®).

[0041] The term “AgLowtumor cells” refers to cells or tumors that do not have abnormally high levels of the tumor antigen (Ag). For example, a breast cancer that does not have abnormally high levels of the tumor antigen HER2 protein can be considered HER2-. About 78% of new female breast cancer cases in the United States between 2015 and 2019 are HER2'. HER2-low breast cancer is a new classification that is used to describe cancer where HER2 proteins are present, but there are not enough for the cancer to be considered HER2-positive. About 50% to 60% of breast cancers are actually HER2-low breast cancers, according to the National Cancer Institute (NCI). These HER2-low cancers have traditionally been classified as HER2-negative (HER2‘). As used throughout this disclosure, the words HER2 and HER2 may be used interchangeably, regardless of uppercase or lower case. In this disclosure, normal cells (non-tumor cells), tumors and tumor cells without abnormally high levels of a tumor antigen (Ag) are considered AgLownormal cells (normal cells) and AgLowtumor cells, respectively.

[0042] Even if a cancer is considered to be HER2-negative, it may still be estrogenpositive or progesterone-positive. Whether or not it is hormone-positive also affects your treatment options.

[0043] The term “Nano-ADC” refers to the nanocomposition comprising an antibody-drug conjugate disclosed herein.

[0044] The term “linker” refers to a molecule that links two or more other molecules. A linker can be a stable (also referred to as a non-cleavable linker, or non-enzymatically cleavable linker), cleavable (as defined hereafter), or a combination thereof. In some cases, a lysine residue, or a lysine-valine dipeptide residue links a coupler MC or SMCC and a PBA can be a non-cleavable linker.

[0045] The term “cleavable linker,” “c-linker,” or a grammatical variation thereof refers to a linker that can be cleaved under proper conditions. As used herein, a cleavable linker can comprise enzymatically-cleavable linkers, acid cleavable linkers that can be cleaved under low pH conditions, acid sensitive hydrazone linker, Glutathione (GSH) cleavable linkers including disulfide linkers, such as SPDB glutathione cleavable linker, that can be cleaved by GSH such as in cytoplasm, Fe(ll) cleavable linkers that can be cleaved by elevating levels of ferrous iron, Cathepsin cleavable linkers that can be cleaved by cathepsin such as in lysosomes, Glycosidase cleavable linkers that can be cleaved by p- glucuronidase such as in lysosomes, Phosphatase cleavable linkers that can be cleaved by phosphatase and pyrophosphates such as in lysosomes, Sulfatase cleavable linkersthat can be cleaved by sulfatase such as in lysosomes, Photo-responsive cleavable linkers that can be cleaved by irradiation with NIR light (such as A = 650-900 nm), Bioorthogonal cleavable linkers that can be cleaved by the bioorthogonal cleavage pair such as Cu(l)-BTTAA / dsProc, a tumor microenvironment activable linker (TMALIN®), an Ortho Hydroxy-Protected Aryl Sulfate (OHPAS) linker, an enzymatically-cleavable peptide linker, a dipeptide linker, a valine— citrulline linker (VC linker), a valine-alanine linker, a VAPAB linker (Valine-alanine-p-aminobenzyl), L-Val-L-Ala-para-aminobenzyl alcohol (VAPAB-OH) linker (“VAPAB linker” and (VAPAB-OH) linker are herein collectively referred to as “VAPAB linker”), a VCPAB (Valine-citrulline-p-aminobenzyl) linker, L-Val-L-Cit-para-aminobenzyl alcohol (VCPAB-OH) linker (VCPAB linker and VCPAB-OH are collectively referred to as “VCPAB linker”), a polypeptide linker, Gly-Gly- L-Phe-N-[(carboxymethoxy)methyl]Glycinamid (GGFG-NCM) linker, a GGFG (Glycyl glycyl-L-phenylalanyl glycyl) linker, a glutamic acid-glycine-citrulline (EGCit, or Glu-Gly- Cit) linkers, a tripeptide linker, a glutamic acid— valine— citrulline (Glu-Val-Cit) linker, p- aminobenzyloxycarbonyl (PABC) linker, p-aminobenzyloxycarbonyl (PABC) linker, or a combination thereof. In some cases, a cleavable linker comprises an enzymatically- cleavable peptide linker, such as Gly-Gly-L-Phe-N-[(carboxymethoxy)methyl]Glycinamid (GGFG-NCM) linker, L-Val-L-Ala-para-aminobenzyl alcohol (VAPAB-OH) linker (herein referred to as “VAPAB linker”), L-Val-L-Cit-para-aminobenzyl alcohol (VCPAB-OH) linker, glutamic acid-glycine-citrulline (EGCit) linkers, acid sensitive hydrazone linker, a linker comprising disulfides, a thioether linker, a glutathione-sensitive disulfide linker, a linker containing a sugar molecule or moiety, a glucuronide containing linker, a glycosidase cleavable linker, a phosphatase cleavable linker, an esterase cleavable linker, a hydrolysis cleavable linker, a modified monodispersed PEG linker, a modified PEO linker, a modified poly-PEG linker, a dipeptide linker, a tripeptide linker, a vali ne— citrulline linker, a glutamic acid— valine— citrulline (Glu-Val-Cit) linker, a valine-alanine linker, a VAPAB linker (Valine-alanine-p-aminobenzyl), a VCPAB (Valine-citrulline-p-aminobenzyl) linker, a glutamic acid-glycine-citrulline (EGCit, or Glu-Gly-Cit) linker, a polypeptide linker, a GGFG (Glycyl glycyl-L-phenylalanyl glycyl) linker, p-aminobenzyloxycarbonyl (PABC) linker, or a combination thereof. Various linkers comprising glycine-glycine- phenylalanine-glycine, such as Gly-Gly-L-Phe-N-[(carboxymethoxy)methyl]Glycinamid (GGFG-NCM) linker or other forms glycine-glycine-phenylalanine-glycine (GGFG) linkers, are collectively referred to as a “GGFG linker”. Various linkers comprising valine and alanine, such as the aforementioned L-Val-L-Ala-para-aminobenzyl alcohol (VAPAB-OH) linker or other forms of valine and alanine linkers, are collectively referred to as a “VAPABlinker” or“VAPAB”. In some cases, a PEOX polymer comprising disulfide bonds can also be suitable as a cleavable linker. The terms “modified monodispersed PEG linker,” “modified PEO linker,” or “modified poly-PEG linker” refers to a corresponding polymer linker that has been modified to comprise one or more cleavable bounds, such as one or more -S-S- bonds or other cleavable bounds disclosed herein. The terms “linker,” “cleavable linker,” “GGFG Linker,” “GGFG,” “VAPAB linker,” or“VAPAB” can also be used to describe a corresponding linker in a reacted form, i.e. , a linker residue that is reacted and incorporated into an ADC, in text or in a drawing. The term “polymer,” “PEG,” “PEOX,” or a variation thereof, can also be used to describe a corresponding polymer in a reacted form, i.e., a polymer that is reacted and incorporated into an ADC, in text or in a drawing.

[0046] The term “alcohol protecting group” refers to hydroxyl (OH) protecting groups in organic synthesis and can include Acetyl (Ac), Benzoyl (Bz), Benzyl (Bn, Bnl), p- Methoxyethoxymethyl ether (MEM), Dimethoxytrityl, [bis-(4- methoxyphenyl)phenylmethyl] (DMT), Methoxymethyl ether (MOM), Methoxytrityl [(4- methoxyphenyl)diphenylmethyl, MMT), p-Methoxybenzyl ether (PMB), Methylthiomethyl ether, Pivaloyl (Piv), Tetrahydropyranyl (THP), Trityl (triphenylmethyl, Tr), Silyl ether such as trimethylsilyl (TMS), tert-butyldimethylsilyl (TBDMS), tri-iso-propylsilyloxymethyl (TOM), and triisopropylsilyl (TIPS) ethers), and Methyl Ethers, Ethoxyethyl ethers.

[0047] The term “amine protecting group” refers to one or more groups protecting amine in organic synthesis and can include Carbobenzyloxy (Cbz) group, p-Methoxybenzyl carbonyl (Moz or MeOZ) group, tert-Butyloxycarbonyl (BOC) group (Common in solid phase peptide synthesis), 9-Fluorenylmethyloxycarbonyl (FMOC) group, Acetyl (Ac) group commonly used in oligonucleotide synthesis for protection of N4 in cytosine and N6 in adenine nucleic bases, Benzoyl (Bz) group commonly used in oligonucleotide synthesis for protection of N4 in cytosine and N6 in adenine nucleic bases, Benzyl (Bn) group, Carbamate group, p-Methoxybenzyl (PMB), 3,4-Dimethoxybenzyl (DMPM), p- methoxyphenyl (PMP) group, Tosyl (Ts) group, and other Sulfonamides (Nosyl & Nps) groups.

[0048] The term “carbonyl protecting group” refers to one or more groups protecting carbonyl in organic synthesis and can include Acetals and Ketals, Acylals, and Dithianes.

[0049] The term “carboxylic acid protecting group” refers to one or more groups protecting carboxylic acid in organic synthesis and can include Methyl esters, Benzyl esters, tert- Butyl esters, Silyl esters, Orthoesters, and Oxazoline.

[0050] The term “thiol protecting group” refers to one or more groups protecting thiol groups in organic synthesis and can include Trityl (Trt), Acetamide methyl (Acm), O-nitrobenzyl (ONB, a photosensitive protecting group that absorbs light at 365 nm), Xanthate (a thiol protecting group used in radical polymerization), Dithiocarbamate (a thiol protecting group used in radical polymerization), Thioacetate (a thiol protecting group used in radical polymerization), and Hmboff / on (a switchable thiol protecting group that can be used in protein chemical synthesis).

[0051] Each of the protecting groups disclosed herein can be removed by a commonly known method.

[0052] The term “pharmaceutical suitable carrier,” “pharmaceutical suitable carriers,” “pharmaceutically suitable carrier,” or “pharmaceutically suitable carriers” refers to one or more inactive ingredients that are in approved drug products. Inactive ingredients listed in the database “Inactive Ingredients in Approved Drug Products” maintained and updated by US FDA can be suitable. In some cases, a pharmaceutical suitable carrier can also be referred to as an excipient.

[0053] The term “subject” or “subjects” used throughout this disclosure refers to an animal, a human or a human patient. The term “animal” refers to wild animals, captured or zooraised animals and domesticated animals including live stocks, farm animals, pets, laboratory animals, such as horse, cattle, pig, donkey, mule, camel, goat, sheep, monkey, rabbit, dog, cat, mouse, rat, and the like. Warm-blooded animals are suitable. The term “human” refers to a human patient having one or more diseases in need of a treatment, a person having one or more medical conditions unrelated to a treatment, or a healthy person. In some cases, a subject can be a human patient or a healthy person.

[0054] The term “antibody,” “antibodies,” or “fragment of an antibody” can include natural or synthetic antibodies that selectively bind to an antigen. The term includes polyclonal and monoclonal antibodies produced from animals, cells including eukaryotic or prokaryotic cells, cell free systems, or chemical synthesis. In addition to intact immunoglobulin molecules, also included in the term “antibodies” are fragments or polymers of those immunoglobulin molecules, and human or humanized versions of immunoglobulin molecules that selectively bind a target antigen.

[0055] The term “aqueous solution” or “aqueous solutions” used throughout this disclosure refers to a solution comprises in a range of from 80% to 100% water, percentage based on the total non-solid weight of the aqueous solution. An aqueous solution can further comprise additional components, such as salt, acid, base, buffer, solvent, organic solvent, particles, emulsion, solids or non-solids, detergents, small molecules, large molecules, other ingredients, or a combination thereof. The term “non-solid weight” refers to the weight from solid contents after the aqueous solution is dried out, such as by removing all the water or other liquids.

[0056] As used herein, the term “isomer” or “isomers” refers to molecules that share the same chemical formula but have their atoms connected differently or arranged differently in space, including structural isomers having respective atoms bonded together in different orders, geometric isomers having atoms bonded in the same order, but differ in the configuration around the bonds, such as cis- ortrans-isomers and enantiomers having the same chemical structure but differ in three-dimensional arrangements of atoms around asymmetric carbon, such that they are mirror images of one another.

[0057] The term “isotope” or “isotopes” refers to different forms of a chemical element which have the same number of protons but a different number of neutrons in their atoms. An isotope can have different physical properties but the same chemical ones. The term also includes radioactive or non-radioactive isotopes. All compounds disclosed herein can comprise one or more isotopes. Some non-limiting examples of isotopes can include deuterium (2H, also referred to as “D” in formulas disclosed herein), tritium (3H), isotoiodine-125 (125l), carbon-12 (12C), carbon-13 (13C), carbon-14 (14C),14N,15N,16O,17O,18O,31P,32P,32S,33S,34S,36S,35S and others, and a combination thereof. Throughout this disclosure, all isotopic variations of the compounds of the present disclosure, whether radioactive or not, are encompassed within the scope of the present disclosure. In some cases, the isotope deuterium (D) can provide structural advantages over H by locking a chiral center in a specific configuration for desired biological activities or pharmaceutical efficacy. In some cases, D can replace H in one or more chiral centers of the compounds disclosed herein. The term “chiral center” refers to an atom covalently linked to four nonidentical substituents, i.e., groups, so it has a nonsuperimposable mirror image. The term "chiral center" can also be referred to as a chirality center.

[0058] The term “C1 -C100 saturated or unsaturated hydrocarbon” refers to a saturated or unsaturated linear, branched or cyclic hydrocarbon having 1-100 carbon atoms and can have one or more hetero-element substitutions. It is understood that a cyclic hydrocarbon shall contain at least 3 carbon atoms or a combination of carbon and hetero-element atoms. Examples of hydrocarbon can include saturated or unsaturated alkyl, substituted alkyl, cycloalkyl, substituted cycloalkyl, aryl, substituted aryl, and a combination thereof.

[0059] In some cases, this disclosure is directed to a modified exatecan (herein referred to as “mExa” throughout this disclosure).

[0060] In some cases, this disclosure is directed to a poly(2-ethyloxazoline) (PEOX) or a PEOX derivative modified exatecan having a formula:a pharmaceutically acceptable salt thereof, a solvate thereof, a prodrug thereof, an isomer thereof, an isotope derivative thereof, and a combination thereof; wherein, v and w each is an integer, v=1-100; w=1-1000;R, R3, R4, R3’, R4’, Rs, Re, R7, Rs and R15 each is independently H, D, C1-C100 saturated or unsaturated hydrocarbon, C1 -C100 saturated or unsaturated hydrocarbon having at least one D, C1 -C100 saturated or unsaturated hydrocarbon having at least one heteroatom, C1-C100 saturated or unsaturated hydrocarbon having at least one heteroatom and at least one D, aromatic group, substituted aromatic group, or a combination thereof;R9 is selected from H, D, C1-C100 saturated or unsaturated hydrocarbon, C1-C100 saturated or unsaturated hydrocarbon having at least one D, C1-C100 saturated or unsaturated hydrocarbon having at least one heteroatom, C1-C100 saturated or unsaturated hydrocarbon having at least one heteroatom and at least one D, OH, NH2, Cl, F, I and Br;R10 is selected from H, Cl, F, I, Br and methoxy;R11 is OH, OR12, SH, SR13, NH2, NHD, ND2, NRuR’u, wherein R12 is an alcohol protecting group, R13 is a thiol protecting group, and Ru and R’14 each is independently H, D, or an amine protecting group; and the heteroatom is selected from N, O, S, P, Cl, Br, I and a combination thereof.

[0061] In some cases, the mExa of formula (1 ) can have v=1 , w=1 and R is selected from methyl, ethyl, benzyl, CNCH2-, cyclopropyl, benzene, 1 ,1 ,1 -trifluoroethyl, n-propyl, and a combination thereof. In some cases, w=1-10, 1-20, 1-30, 1 -40, 1-50, 1-60, 1-70, 1-80, or 1-90. In some cases, R, R3, R4, R3’, R4’, Rs, Re, R7, Rs, and R15 each is independently H, D, C1 -C22 saturated or unsaturated hydrocarbon, C1-C22 saturated or unsaturated hydrocarbon having at least one D, C1-C22 saturated or unsaturated hydrocarbon having at least one heteroatom, C1 -C22 saturated or unsaturated hydrocarbon having at leastone heteroatom and at least one D, aromatic group, substituted aromatic group, or a combination thereof. In some cases, R3, R4, R3’, R4’, R5, R6, R7, R8, and R15 each is independently H or D. In some cases, R9 is a methyl. In some cases, R10 is F. In some cases, R11 is OH. In some cases, the mExa of formula (1 ) can have a combination of the groups disclosed hereabove.

[0062] In some cases, this disclosure is directed to a modified exatecan (mExa) having a formula:a pharmaceutically acceptable salt thereof, a solvate thereof, a prodrug thereof, an isomer thereof, an isotope derivative thereof, and a combination thereof; wherein,R’, R”, R3, R4, R3’, R4 , R5, R6, R7, and Rs each is independently H, D, C1-C100 saturated or unsaturated hydrocarbon, C1 -C100 saturated or unsaturated hydrocarbon having at least one D, C1 -C100 saturated or unsaturated hydrocarbon having at least one heteroatom, C1-C100 saturated or unsaturated hydrocarbon having at least one heteroatom and at least one D, aromatic group, substituted aromatic group, or a combination thereof;R9 is selected from H, D, C1-C100 saturated or unsaturated hydrocarbon, C1-C100 saturated or unsaturated hydrocarbon having at least one D, C1-C100 saturated or unsaturated hydrocarbon having at least one heteroatom, C1-C100 saturated orunsaturated hydrocarbon having at least one heteroatom and at least one D, OH, NH2, Cl, F, I and Br;R10 is selected from H, Cl, F, I, Br and methoxy;R11 is OH, OR12, SH, SR13, NH2, NHD, ND2, NRuR’-u, wherein R12 is an alcohol protecting group, R13 is a thiol protecting group, and Ru and R’14 each is independently H, D, or an amine protecting group; the heteroatom is selected from N, O, S, P, Cl, Br, I and a combination thereof; and X is O, S, or NH.

[0063] In some cases, the mExa of formulas (2) or (3) each can independently have R’, R”, R3, R4, R3’, R4’, Rs, Re, R7, and Rs each is independently H or D. In some cases, R’, R”, R3, R4, Rs’, R4’, Rs, Re, R7, and Rs each is independently H, D, C1-C22 saturated or unsaturated hydrocarbon, C1-C22 saturated or unsaturated hydrocarbon having at least one D, C1-C22 saturated or unsaturated hydrocarbon having at least one heteroatom, C1-C22 saturated or unsaturated hydrocarbon having at least one heteroatom and at least one D, aromatic group, substituted aromatic group, or a combination thereof.

[0064] In some cases, Rg is methyl. In some cases, R10 is F. In some cases, R11 is OH. In some cases, the mExa of formula (1 ) can have a combination of the groups disclosed hereabove.

[0065] In some cases, an mExa can have a formula selected from Formula ID 1 , 1-1 , 1- 2, 2, 2-1 , 2-2, 3, 3-1 , 3-2, 1001 -1135 or a combination thereof as disclosed hereafter.

[0066] Any of the alcohol protecting groups, thiol protecting groups, or amine protecting groups disclosed above can be suitable. In some cases, an mExa can comprise an alcohol protecting group R12. In some cases, an mExa can comprise a thiol protecting group R13. In some cases, an mExa can comprise an amine protecting group Ri or R’i4.

[0067] In some cases, this disclosure is directed to a bioactive composition comprising a payload bioactive agent (PBA) comprising a formula selected from Formula ID 1 , 2, 3:(1 ),a pharmaceutically acceptable salt thereof, a solvate thereof, a prodrug thereof, an isomer thereof, an isotope derivative thereof, and a combination thereof; wherein, v and w each is an integer, v=1-100; w=1 -1000;R, R’, R”, R3, R4, R3’, R4’, Rs, Rs, R7, Rs and R15 each is independently H, D, CICI 00 saturated or unsaturated linear or C3-C100 cyclic hydrocarbon, C1 -C100 saturated or unsaturated linear or C3-C100 cyclic hydrocarbon having at least one heteroatom, CICI 00 saturated or unsaturated linear or C3-C100 cyclic hydrocarbon having at least one heteroatom and at least one D, aromatic group, substituted aromatic group, or a combination thereof;R9 is selected from H, D, C1-C100 saturated or unsaturated linear or C3-C100 cyclic hydrocarbon, C1 -C100 saturated or unsaturated linear or C3-C100 cyclic hydrocarbon having at least one heteroatom, C1-C100 saturated or unsaturated linear or C3-C100 cyclic hydrocarbon having at least one heteroatom and at least one D, OH, NH2, Cl, F, I and Br;R10 is selected from H, Cl, F, I, Br and methoxy;R11 is OH, OR12, SH, SR13, NH2, NHD, ND2, NRuR’u, wherein R12 is an alcohol protecting group, R13 is a thiol protecting group, and Ru and R’14 each is independently H, D, or an amine protecting group; the heteroatom is selected from N, O, S, P, Cl, Br, I and a combination thereof; andX is O, S, or NH.

[0068] In some cases, v is in a range of from 1 -100, 1-90, 1 -80, 1-70, 1-60, 1-50, 1-40, 1-30, 1-20, 1-18, 1-16, 1-14, 1-12, 1-10, 1-8, 1-6, 1-4, 1-3, or 1-2 and w is in a range of from 1 - 1000. In some cases, v is 1. In some cases, w is in a range of from 1- 1000, 1-900, 1-800, 1-700, 1 -600, 1-500, 1-400, 1-300,1-200, 1-150, 1-120, 1-100, 1-80, 1-70, 1-60, 1-50, 1-40, 1-30, 1-20, 1-18, 1-16, 1-14, 1-12, 1 -10, 1-8, 1-6, 1-4, 1-2, and v is in arrange of from 1-100. In some cases, w is 1-200. In some cases, w is in a range of from 1 -1000, 1-900, 1-800, 1-700, 1-600, 1-500, 1-400, 1-300, 1-200, 1-180, 1-160, 1-140, 1-120, 1-100, 1-80, 1-60, 1-40, 1-30, 1- 20, 1-10, or 1-2 and v is in a range of from1 - 100. In some cases, w is 1-20. In some cases, v is 1 and w is 1-20, 1-10, 1 -8, 1 -6, 1 -4, or 1-2. In some cases, v and w are in a combination of the ranges disclosed above.

[0069] In some cases, X is O. In some cases, X is S. In some cases, X is NH.

[0070] In some cases, R, R’ and R” each is H, C1 -22 alkyl group, C1-20 alkyl group, CI- 18 alkyl group, C1-16 alkyl group, C1-14 alkyl group, C1-12 alkyl group, C1-10 alkyl group, C1-8 alkyl group, C1-6 alkyl group, aromatic group, or substituted aromatic group. In some cases, at least one of the R, R’ and R” is C1-22 alkyl group, C1-20 alkyl group, C1-18 alkyl group, C1 -16 alkyl group, C1-14 alkyl group, C1 -12 alkyl group, C1-10 alkyl group, C1-8 alkyl group, or C1-6 alkyl group. In some cases, at least one of the R, R’ and R” is C22 alkyl group, C20 alkyl group, C18 alkyl group, C16 alkyl group, C14 alkyl group, C12 alkyl group, C10 alkyl group, or C8 alkyl group, providing hydrophobicity to the modified exatecan. In some cases, at least one H is D.

[0071] In some cases, at least one of the R, R’ and R” is C1-22 alkyl group.

[0072] In some cases, v=1 , w=1 , and R is selected from methyl, ethyl, benzyl, CNCH2-, cyclopropyl, benzene, 1 ,1 ,1 -trifluoroethyl, n-propyl, and a combination thereof. In some cases, R, R’, R”, R3, R4, R3’, R4’, Rs, Re, R7, Rs and R15 each is independently H, D, CI- 022 saturated or unsaturated hydrocarbon, C1-C22 saturated or unsaturated hydrocarbon having at least one D, C1-C22 saturated or unsaturated hydrocarbon having at least one heteroatom, C1 -C22 saturated or unsaturated hydrocarbon having at least one heteroatom and at least one D, aromatic group, substituted aromatic group, or a combination thereof. In some cases, R3, R4, R3’, R4’, Rs, Re, R7, Rs and R15 each is independently H or D. In some cases, R9 is a methyl. In some cases, R10 is F. In some cases, R11 is OH. In some cases, the PBA of Formula ID 1 , 2 or 3 can have a combination of the groups disclosed hereabove.

[0073] Any of the alcohol protecting groups, thiol protecting groups, or amine protecting groups disclosed above can be suitable.

[0074] In some cases, the bioactive composition can comprise a PBA comprising a Formula ID 1-1 , wherein R3, R4, Rs, Rs, Rz, Rs can be H, R9 can be CH3, R10 can be F, R11 is OH:wherein, v=1-100; w=1-1000; and R= H, D, 01-22 alkyl group, aromatic group, or substituted aromatic group.

[0075] In some cases, v=1 , w=1 , and the bioactive composition can comprise a PBA comprising a Formula ID 1-2:1 -2.

[0076] In some cases, the bioactive composition can comprise a PBA comprising a Formula ID 2-1 , wherein R3, R4, R3’, R4’, Rs, Rs, R7, and Rs each is H, R’= H, D, C1-22 alkyl group, aromatic group, or substituted aromatic group, R9 can be CH3, R10 can be F, R11 is OH,

[0077] In some cases, X is NH and the bioactive composition can comprise a PBA comprising a Formula ID 2-2:

[0078] In some cases, the bioactive composition can comprise a PBA comprising a Formula ID 3-1 , wherein R3, R4, R3’, R4’, Rs, Re, R , and Rs each is H, R”= H, D, C1-22 alkyl group, aromatic group, or substituted aromatic group, R9 can be CH3, R10 can be F, R11 is OH:

[0079] In some cases, X is NH and the bioactive composition can comprise a PBA comprising a Formula ID 3-2:

[0080] In some cases, X is NH, R” is H, and the bioactive composition can comprise a PBA comprising a Formula ID 3-3:

[0081] In some cases, the PBA can have a formula selected from Formula ID 1 , 1-1 , 1-2, 2, 2-1 , 2-2, 3, 3-1 , 3-2, 1001-1 135, a pharmaceutically acceptable salt thereof, a solvate thereof, a prodrug thereof, an isomer thereof, an isotope derivative thereof, and a combination thereof.

[0082] In some cases, the PBA can have a formula selected from Formula ID 1001-1135, a pharmaceutically acceptable salt thereof, a solvate thereof, a prodrug thereof, an isomer thereof, an isotope derivative thereof, and a combination thereof:

[0083] In some cases, Formulas 1025, 1054, 1083 or 1112 each can comprise 3 As being C, 2 As being C and 1 A being N, or 1 A being C and 2 As being N; and Formulas 1026, 1055, 1084 or 1113 each can comprise 3 As being C, 2 As being C and 1 A being N, 1 A being C and 2 As being N, or 3 As being N.

[0084] In some cases, Formulas 1025, 1054, 1083 or 1112 each can comprise 3 As being C. In some cases, Formulas 1025, 1054, 1083 or 1112 each can comprise 2 As being C and 1 A being N. In some cases, Formulas 1025, 1054, 1083 or 1112 each can comprise 1 A being C and 2 As being N. In some cases, Formulas 1025, 1054, 1083 or 1112 each can comprise 3 As being C. In some cases, Formulas 1025, 1054, 1083 or 1112 each can comprise 2 As being C and 1A being N. In some cases, Formulas 1025, 1054, 1083 or 1112 each can comprise 1 A being C and 2 As being N. In some cases, Formulas 1025, 1054, 1083 or 1112 each can comprise 3 As being N.

[0085] In some cases, the bioactive composition can be a bioactive agent (BA) or a PBA. In some cases, the bioactive composition can be a PBA in an antibody-drug conjugate (ADC). In some cases, the bioactive composition can be a PBA in a Nano-ADC.

[0086] In some cases, the bioactive composition can further comprise a polymer, a linker, optionally a coupler, or a combination thereof, wherein the PBA can be covalently linked to the polymer, the linker, or optionally, the coupler, wherein, the polymer can comprise a peptide, a polyethylene glycol (PEG), polyoxazoline (POX), or a combination thereof, wherein the polyoxazoline (POX) comprises a linear portion, a branched portion, or a combination thereof, and wherein the polyoxazoline (POX) comprises poly(2-methyloxazoline), poly(2-ethyloxazoline), poly(2- propyloxazoline), poly(2-isopropyloxazoline), poly(2-substituted oxazoline), or a combination thereof; andthe coupler can be selected from succinimidyl-4-(N- maleimidomethyl)cyclohexane-1 -carboxylate (SMCC), maleimide (Mai), maleimidocaproyl (MC), maleimidomethyl cyclohexane-1 -carboxylate (MCC), dibromomaleimide (DBM), glycan-based coupler, N-acetylglucosamine (GlcNAc)-based coupler, and a combination thereof.

[0087] In some cases, polymer can comprise a single amino acid residue, dipeptide, tripeptide, or polypeptide. In some cases, as mentioned above, the polymer can be a peptide comprising in a range of from 1 to 100 amino acid residues. In some cases, a polymer can comprise a lysine, glutamic acid, glutamine, arginine, valine, or a combination thereof.

[0088] In some cases, the linker can comprise a cleavable linker that can comprise an enzymatically-cleavable linker, acid cleavable linkers, acid sensitive hydrazone linker, a glutathione (GSH) cleavable linker, a linker comprising disulfides, a SPDB glutathione cleavable linker, a thioether linker, a glutathione-sensitive disulfide linker, a Fe(ll) cleavable linker, a Cathepsin cleavable linker, a linker containing a sugar molecule or moiety, a glucuronide containing linker (such as a P-Glucuronide Linker), a glycosidase cleavable linker, a phosphatase cleavable linker, a sulfatase cleavable linker, an esterase cleavable linker, a hydrolysis cleavable linker, a Photo-responsive cleavable linker, a Bioorthogonal cleavable linker, a tumor microenvironment activable linker (TMALIN®), an Ortho Hydroxy-Protected Aryl Sulfate (OHPAS) linker, a modified monodispersed PEG linker, a modified PEG linker, a modified poly-PEG linker, an enzymatically-cleavable peptide linker, a dipeptide linker, a valine— citrulline linker (VC linker), a valine-alanine linker, a VAPAB linker (Valine-alanine-p-aminobenzyl), L-Val-L-Ala-para-aminobenzyl alcohol (VAPAB-OH) linker, a VCPAB (Valine-citrulline-p-aminobenzyl) linker, L-Val-L- Cit-para-aminobenzyl alcohol (VCPAB-OH) linker, a polypeptide linker, Gly-Gly-L-Phe-N- [(carboxymethoxy)methyl]Glycinamid (GGFG-NCM) linker, a GGFG (Glycyl glycyl-L- phenylalanyl glycyl) linker, a glutamic acid-glycine-citrulline (EGCit, or Glu-Gly-Cit) linkers, a tripeptide linker, a glutamic acid— valine— citrulline (Glu-Val-Cit) linker, p- aminobenzyloxycarbonyl (PABC) linker, p-aminobenzyloxycarbonyl (PABC) linker, or a combination thereof;

[0089] It is understood that the PBA is in a reacted form wherein the PBA is covalently linked to the polymer, the cleavable linker, or the coupler.

[0090] In some cases, the polymer can comprise polyoxazoline (POX).

[0091] In some cases, a PEOX polymer can be functionalized to have one or more desired functional groups as the first terminal group and the second terminal group, respectively.In some cases, functionalized PEOX polymer can facilitate the formation of polymer-PBA- albumin aggregates in vitro, in blood stream, in plasma, or in cytosol. In some cases, the PEOX polymer can be functionalized to comprise disulfide bonds and covalent linkage of multiple drug molecules, such as two or more PBAs, such as exatecan (Exa), modified exatecan (mExa), or a combination thereof, to the polymer.

[0092] In some cases, a functionalized PEOX polymer can comprise one or more end groups (EG). In some cases, the end group can be the first terminal group (R1 ), the second terminal group (R2), or a combination thereof. In some cases, a functionalized PEOX polymer can comprise one or more end groups (EG) -H, -NH2, -CH2N3, -CH2CH2N3, -CH2CH2CH2N3, -NHBoc, -NHFmoc, or a combination thereof. In some cases, a functionalized PEOX polymer can comprise one or more end groups (EG) selected from the group consistent of-H, -NH2, -NHCH2NH3, -NHCH2CH2NH3, -NHCH2CH2CH2NH3, -CH2N3,-CH2CH2N3, -CH2CH2CH2N3, -NHBoc, -NHFmoc, -OH, -N3, and a combination thereof. In some cases, a functionalized PEOX polymer can comprise a first terminal group comprising one or more end groups (EG) selected from -H, -NH2, -NHCH2NH3, -NHCH2CH2NH3, -NHCH2CH2CH2NH3, -CH2N3, -CH2CH2N3, -CH2CH2CH2N3 and a combination thereof. In some cases, a functionalized PEOX polymer can comprise a second terminal group selected from -NHBoc, -NHFmoc, -OH, -N3, and a combination thereof. In some cases, a functionalized PEOX polymer can comprise one or more end groups (EG) having a formulaor a combination thereof. In some cases, a PEOX polymer can be functionalized and can have multiple PBAs, such as drug molecules, linked thereon at one or more of the second terminals of the PEOX polymer via one or more linkers. In some cases, a functionalized PEOX can comprise two PBAs, such as Exatecan, linked to one PEOX polymer at one or more of the second terminals via one or more linkers. In some cases, a functionalized PEOX can comprise four PBAs, such as Exatecan, linked to one PEOX polymer at one or more of the second terminals via linkers L2.

[0093] In some cases, the R1 group can a group that can react with a coupler selected from Formula ID 201-209 as described hereafter. In some cases, the R2 group can be a group that can react with a functional group of the Formula ID 1 , 1-1 , 1-2, 2, 2-1 , 2-2, 3, 3-1 , 3-2, and 1001-1 135. In some cases, the R2 group can be a group that can react with a functional group of a linker disclosed herein. In some cases, the R2 group can be agroup that can react with a functional group of a cleavable linker disclosed herein, such as Formula ID 101-141.

[0094] In some cases, the at least two PBAs can be selected from Formula ID 1 , 1-1 , 1- 2, 2, 2-1 , 2-2, 3, 3-1 , 3-2, and 1001-1135, a pharmaceutically acceptable salt thereof, a solvate thereof, a prodrug thereof, an isomer thereof, an isotope derivative thereof, and a combination thereof.

[0095] In some cases, the bioactive composition can comprise at least a first PBA covalently linked to a first polymer and a second PBA covalently linked to a second polymer, wherein the first polymer and the second polymer can be the same or different, wherein the first PBA and the second PBA can be the same or different and each of the at least two different PBAs can be selected from same or different natural or synthetic small molecule-based drugs, inorganic-based drugs, toxin molecules having cytotoxicity, molecules having cytotoxicity to target cells, derivatives thereof, or a combination thereof. In some cases, the first and the second PBAs can be independently selected from Formula ID 1 , 1-1 , 1-2, 2, 2-1 , 2-2, 3, 3-1 , 3-2, and 1001-1135, one or more additional cytotoxic agents disclosed herein, a pharmaceutically acceptable salt thereof, a solvate thereof, a prodrug thereof, an isomer thereof, an isotope derivative thereof, and a combination thereof.

[0096] In some cases, the bioactive composition can comprise branched polyoxazoline.

[0097] In some cases, the bioactive composition can comprise a polymer comprising a side chain that comprises at least an amino acid comprising lysine, arginine, glutamic acid, glutamine, or a combination thereof, polyoxazoline (POX), poly(2-methyloxazoline), poly(2-ethyloxazoline), poly(2-propyloxazoline), poly(2-isopropyloxazoline), poly(2- substituted oxazoline), or a combination thereof.

[0098] In some cases, the bioactive composition can comprise at least one polymer side chain attached to the polymer. In some cases, the polymer side chain can be free from any PBAs. In some cases, the bioactive composition can comprise one or more polymer side chains that at least one of the side chains can be attached to one or more PBAs. In some cases, the bioactive composition can comprise one or more polymer side chains that each of the side chains can be attached to one or more PBAs.

[0099] In some cases, the polymer can comprise polyoxazoline (POX) having a branched portion that can comprise two or more branches, and wherein at least one of the two or more branches can be the polymer side chain and can be free from the PBA. In some cases, each of the two or more branches can be the polymer side chain and can be attached to one or more PBAs.

[0100] The polymer side chain can comprise the PEOX polymer, the PEG polymer, or a combination thereof. In some cases, a polymer side chain can comprise any of the PEOX polymers disclosed herein. In some cases, a polymer side chain can comprise any of the PEG polymers disclosed herein. In some cases, a polymer side chain can comprise a combination of any of the PEOX polymers and the PEG polymers disclosed herein.

[0101] In some cases, the bioactive composition can comprise at least one polymer side chain attached to the polymer. In some cases, the polymer side chain can be free from any PBAs. In some cases, the bioactive composition can comprise one or more polymer side chains that at least one of the side chains can be attached to one or more PBAs. In some cases, the bioactive composition can comprise one or more polymer side chains that each of the side chains can be attached to one or more PBAs.

[0102] In some cases, the bioactive composition can comprise at least a branched or linear polymer side chain. In some cases, a bioactive composition of this disclosure can comprise a Linker L1 (2a), a Linker L2 (2b) or a combination thereof, without any cleavable linkers between the TBA (1), polymer (4) and first PBA (3), and a branched polymer side chain (4’) attached to the polymer (4) (similar to the one illustrated in FIG. 8A without the TBA or antibody). In some cases, a bioactive composition can comprise at least one cleavable linker, such as the first cleavable linker (8a), the second cleavable linker (8b), or a combination thereof, and a branched polymer side chain (4’) attached to the polymer (similar to the one illustrated in FIG. 8C without the TBA or antibody). In some cases, a bioactive composition can comprise a Linker L1 (2a), a Linker L2 (2b) or a combination thereof, without any cleavable linkers between the TBA (1 ), polymer (4) and first PBA (3), and a linear polymer side chain (4”) attached to the polymer (4) (similar to the one illustrated in FIG. 8E without the TBA or antibody). In some cases, a bioactive composition can comprise at least one cleavable linker, such as the first cleavable linker (8a), the second cleavable linker (8b), or a combination thereof, and a linear polymer side chain (4”) attached to the polymer (4) (similar to the one illustrated in FIG. 8G without the TBA or antibody). R1 and R2 represent the first terminal group and the second terminal group, respectively, as disclosed herein. In some cases, a bioactive composition can comprise at least one linker and a polymer side chain attached to the linker, such as schematically illustrated in FIG. 8I-FIG. 8L. For simplicity, a linear or a branched polymer can be shown in some figure and can represent a linear polymer, a branched polymer or a combination thereof.

[0103] In some cases, the bioactive composition can comprise at least a branched or linear polymer side chain that is attached to at least a PBA. In some cases, a bioactivecomposition of this disclosure can comprise a Linker L1 (2a), a Linker L2 (2b) or a combination thereof, without any cleavable linkers between the TBA (1 ), polymer (4) and first PBA (3), and a branched polymer side chain (4’) attached to the polymer (4) and at least a second PBA (3’) (similar to the one illustrated in FIG. 9A without the TBA or antibody). In some cases, a bioactive composition can comprise at least one cleavable linker, such as the first cleavable linker (8a), the second cleavable linker (8b), or a combination thereof, and a branched polymer side chain (4’) attached to the polymer and at least a second PBA (3’) (similar to the one illustrated in FIG. 9C without the TBA or antibody). In some cases, a bioactive composition can comprise a Linker L1 (2a), a Linker L2 (2b) or a combination thereof, without any cleavable linkers between the TBA (1), polymer (4) and first PBA (3), and a linear polymer side chain (4”) attached to the polymer (4) and at least a second PBA (3’) (similar to the one illustrated in FIG. 9E without the TBA or antibody). In some cases, a bioactive composition can comprise at least one cleavable linker, such as the first cleavable linker (8a), the second cleavable linker (8b), or a combination thereof, and a linear polymer side chain (4”) attached to the polymer (4) and at least a second PBA (3’) (similar to the one illustrated in FIG. 9G without the TBA or antibody). In some cases, a bioactive composition can comprise at least one linker and a polymer side chain attached to the linker, such as schematically illustrated in FIG. 9H- FIG. 9L. R1 and R2 represent the first terminal group and the second terminal group, respectively, as disclosed herein. In some cases, the bioactive composition can comprise at least one of the R1 , R2 or a combination thereof of the polymer side chain, free from the PBAs. In some cases, the bioactive composition can comprise at least one of the R1 , R2 or a combination thereof, of the polymer side chain, attached to one or more PBAs, such as the first PBA, the second PBA, or a combination thereof. The first PBA (3) and the second PBA (3’) can be the same or different, and each can be independently selected form the PBAs disclosed herein, or a combination thereof.

[0104] In some cases, the bioactive composition can comprise covalently linked PBA and the polymer (Formula ID 10). In some cases, the bioactive composition can comprise covalently linked PBA and the linker, such as a Linker L2 (Formula ID 1 1 ).

[0105] In some cases, the bioactive composition can comprise covalently linked PBA, the polymer, and the linker, wherein at least one linker can be linked between the polymer and the PBA as shown in Formula ID 12. In some cases, the polymer can be positioned between the linker and the PBA as shown in Formula ID 13. In some cases, the polymer can be positioned between 2 linkers, such as a Linker L1 and a Linker L2 as shown in Formula ID 14:wherein any of the polymers, the linkers and the PBA disclosed above can be suitable. The linker can comprise a cleavable linker. The Linker L1 , Linker L2, or a combination thereof, each can comprise a cleavable linker. Some unlimited examples are schematically illustrated in FIG. 1A-FIG. 1C, FIG. 2A-FIG 2E, FIG. 3A-FIG. 3D, FIG. 4A- FIG. 4D, FIG. 5, FIG. 6A-FIG. 6C and FIG. 7A-FIG. 7H. In some cases, Linker L1 and L2 each can be independently a cleavable linker, and can be the same or different. In some cases, Linker L1 and L2 both can be independently a cleavable linker, and can be the same or different. The Linker L1 can be covalently linked to the TBA and covalently linked to the reacted first terminal group of the polymer, the Linker L2 can be covalently linked to the PBA and covalently linked to the reacted second terminal group of the polymer, with a proviso that either Linker L1 or Linker L2 can each be covalently linked to TBA or PBA when the polymer is absent.

[0106] I n some cases, the bioactive composition can further comprise a coupler covalently linked to the polymer, the linker, the PBA, or a combination thereof. In some cases, a coupler is covalently linked to a PBA (Formula ID 21 ). In some cases, the coupler is covalently linked to the polymer and the PBA with the polymer positioned between the coupler and the PBA (Formula ID 22). In some cases, a coupler is covalently linked to at least one linker and PBA with the linker positioned between the coupler and the PBA (Formula ID 23). In some cases, a coupler is covalently linked to at least one linker, a polymer and PBA with the linker positioned between the polymer and the PBA (Formula ID 24). In some cases, a coupler is covalently linked to at least one linker, a polymer and PBA with the linker positioned between the coupler and the polymer (Formula ID 25). In some cases, a coupler is covalently linked to at least two linkers, a polymer and PBA with one linker, such as a Linker L1 positioned between the coupler and the polymer, and at least one linker, such as a Linker L2 positioned between the polymer and the PBA (Formula ID 26). In some cases, the bioactive composition can comprise a covalently linked coupler, polymer, one or more linkers, and PBA as shown in Formula ID 21 , 22, 23, 24, 25, and 26:wherein any of the couplers disclosed hereafter, the polymers, the linkers and the PBA disclosed herein can be suitable.

[0107] In some cases, the linker can comprise a cleavable linker selected from the Formula ID 101-141 , or a combination thereof:

[0108] In some cases, the coupler can comprise cysteine-based coupler, succinimidyl-4- (N-maleimidomethyl)cyclohexane-l -carboxylate (SMCC), maleimide (Mai), dibromomaleimide (DBM), maleimidocaproyl (MC), maleimidomethyl cyclohexane-1- carboxylate (MCC), Mal-PEG, a CL2A linker that is a cleavable complicated PEG8- and triazole-containing PABC-peptide-MC linker that is cleavable through pH sensitivity, giving rise to bystander effect, and binds the antibody at a cysteine residue via a disulfide bond, glycan-based coupler, N-acetylglucosamine (GlcNAc)-based coupler, or a combination thereof.

[0109] ln some cases, the coupler can be selected from Formula ID 201 -209, or a combination thereof:

[0110] In some cases, the bioactive composition can comprise a formula selected from Formula ID 10, 11 , 12, 13, 14, 21 , 22, 23, 24, 25, 26 or a combination thereof; wherein, the coupler, when present, can be selected from Formula ID 201 , 202, 203, 204, 205, 206, 207, 208, 209, and a combination thereof; the polymer can be selected from a peptide, a polyethylene glycol (PEG), polyoxazoline (POX), or a combination thereof, wherein the polyoxazoline (POX) comprises a linear portion, a branched portion, or a combination thereof, and wherein the polyoxazoline (POX) comprises poly(2-methyloxazoline), poly(2-ethyloxazoline), poly(2- propyloxazoline), poly(2-isopropyloxazoline), poly(2-substituted oxazoline), and a combination thereof;the linker can comprise a cleavable linker selected from Formula ID 101 - 141 , and a combination thereof; and the PBA can be selected from Formula ID 1 , 1-1 , 1-2, 2, 2-1 , 2-2, 3, 3-1 , 3-2 and 1001-1135, a pharmaceutically acceptable salt thereof, a solvate thereof, a prodrug thereof, an isomer thereof, an isotope derivative thereof, and a combination hereof.

[0111] In some cases, polymer can comprise a single amino acid residue, dipeptide, tripeptide, or polypeptide. In some cases, as mentioned above, the polymer can comprise a peptide comprising in a range of from 1 to 100 amino acid residues. In some cases, a polymer can comprise a lysine, glutamic acid, glutamine, arginine, valine, or a combination thereof.

[0112] In some cases, the bioactive composition can comprise at least two PBAs that can be the same or different. In some cases, the bioactive composition can comprise at least two different PBAs covalently linked to each polymer, such as each PEOX polymer or each PEG polymer, wherein each of the at least two different PBAs can be selected from at least two different natural or synthetic small molecule-based drugs, inorganic-based drugs, toxin molecules having cytotoxicity, molecules having cytotoxicity to target cells, derivatives thereof, or a combination thereof.

[0113] The polymer side chain can comprise at least an amino acid comprising lysine, arginine, glutamic acid, glutamine, or a combination thereof, the PEOX polymer, the PEG polymer, or a combination thereof. In some cases, a polymer side chain can comprise any of the PEOX polymers disclosed herein. In some cases, a polymer side chain can comprise any of the PEG polymers disclosed herein. In some cases, a polymer side chain can comprise a combination of any of the PEOX polymers and the PEG polymers disclosed herein.

[0114] In some cases, the bioactive composition can comprise at least one polymer side chain attached to the polymer. In some cases, the polymer side chain can be free from any PBAs. In some cases, the bioactive composition can comprise one or more polymer side chains that at least one of the side chains can be attached to one or more PBAs. In some cases, the bioactive composition can comprise one or more polymer side chains that each of the side chains can be attached to one or more PBAs.

[0115] In some cases, the polymer can comprise polyoxazoline (POX) having a branched portion that can comprise two or more branches, and wherein at least one of the two or more branches can be the polymer side chain and can be free from the PBA. In some cases, each of the two or more branches can be the polymer side chain and can be attached to one or more PBAs.

[0116] In some cases, the PBA can comprise an additional cytotoxic agent disclosed herein. In some cases, the PBA can further comprise calicheamicin, DM1 (antimicrotubule agent T-DM1 , a derivative of maytansine), DM4 (Ravtansine), Dxd (deruxtecan), exatecan, IRDye700DX (a near-infrared (NIR) photosensitizer suitable for near infrared (NIR) photoimmunotherapy (PIT)), MMAE (Monomethyl auristatin E, also known as vedotin), MMAF (Monomethyl auristatin F), pseudomonas exotoxin A (a potent exotoxin produced by the bacterium Pseudomonas aeruginosa), pyrrolobenzodiazepine (PBD), SG3199 (a cytotoxic DNA minor groove interstrand crosslinking pyrrolobenzodiazepine (PBD) dimer), SN-38, irinotecan, camptothecin, amanitin (an inhibitor of RNA Polymerase II), duocarmycins (DNA-alkylators), halichondrin (Eribulin, also known Halaven®, under respective trademark), norhalichondrin B, maytansinoids, maitansine (INN), maytansine, PUN-159682 (an inhibitor of DNA Topoisomerase II), tubulysins, tubulysin, pretubulysin (PT), tubulysin A (TubA), tubulysin M, or a combination thereof. In some cases, other PBAs disclosed herein can be suitable.

[0117] In some cases, the bioactive composition can comprise the polymer comprising a peptide, a polyoxazoline (POX), or a combination thereof, wherein the polyoxazoline (POX) can comprise a linear portion, a branched portion, or a combination thereof, and wherein the polyoxazoline (POX) comprises poly(2-methyloxazoline), poly(2- ethyloxazoline), poly(2-propyloxazoline), poly(2-isopropyloxazoline), poly(2-substituted oxazoline), and a combination thereof, polyoxazoline (POX) having a branched portion that can comprise two or more branches, at least one polymer side chain attached to the polymer, and one or more PBAs selected from the PBA having the Formula ID 1 , 1-1 , 1- 2, 2, 2-1 , 2-2, 3, 3-1 , 3-2, and 1001-1 135, calicheamicin, DM1 (antimicrotubule agent T- DM1 , a derivative of maytansine), DM4 (Ravtansine), Dxd (deruxtecan), exatecan, IRDye700DX (a near-infrared (NIR) photosensitizer suitable for near infrared (NIR) photoimmunotherapy (PIT)), MMAE (Monomethyl auristatin E, also known as vedotin), MMAF (Monomethyl auristatin F), pseudomonas exotoxin A (a potent exotoxin produced by the bacterium Pseudomonas aeruginosa), pyrrolobenzodiazepine (PBD), SG3199 (a cytotoxic DNA minor groove interstrand crosslinking pyrrolobenzodiazepine (PBD) dimer), SN-38, irinotecan, camptothecin, amanitin (an inhibitor of RNA Polymerase II), duocarmycins (DNA-alkylators), halichondrin (Eribulin, also known Halaven®, under respective trademark), norhalichondrin B, maytansinoids, maitansine (INN), maytansine, PUN-159682 (an inhibitor of DNA Topoisomerase II), tubulysins, tubulysin, pretubulysin (PT), tubulysin A (TubA), tubulysin M, a pharmaceutically acceptable salt thereof, asolvate thereof, a prodrug thereof, an isomer thereof, an isotope derivative thereof, and a combination thereof.

[0118] In some cases, the bioactive composition can comprise two or more same or different PBAs. In some cases, the bioactive composition can comprise Formula ID 2005, 2027, 2028, 2029, 2030, or a combination thereof.

[0119] In some cases, the bioactive composition can comprise a biomolecule having a formula selected from Formula ID 2001-2040:a pharmaceutically acceptable salt thereof, a solvate thereof, a prodrug thereof, an isomer thereof, an isotope derivative thereof, and a combination hereof; wherein y and y’ each is an integer in a range of from 1 to 100, m and m’ each is an integer in a range of from 2-100.

[0120] In some cases, y and y’ each is independently 1 , 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 , 12,13, 14, 15, 16, 17, 18, 19, 20, 21 , 22, 23 or 24. In some cases, y is 2, 4, 6, 8, 10, 12, 14,16 or 18. In some cases, y’ is 2, 4, 6, 8, 10, 12, 14, 16 or 18.

[0121] ln some cases, m is 1 , 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 , 12, 13, 14, 15, 16, 17, 18, 19,20, 21 , 22, 23 or 24. In some cases, m and m’ each is independently 1 , 2, 4, 6, 8, 12 or 24.

[0122] In some cases, a bioactive composition can comprise at least one linker and a polymer side chain attached to the linker, such as Formula ID 2025, 2026, 2027 or a combination thereof.

[0123] In some cases, this disclosure is directed to a pharmaceutical composition comprising a PBA selected from Formula ID 1 , 2, 3:a pharmaceutically acceptable salt thereof, a solvate thereof, a prodrug thereof, an isomer thereof, an isotope derivative thereof, optionally, an additional cytotoxic agent, or a combination thereof; optionally, a pharmaceutically suitable carrier; wherein, v and w each is an integer, v=1-100; w=1-1000;R, R’, R”, R3, R4, R3’, R4’, Rs, Re, R7, Rs and R15 each is independently H, D, CICI 00 saturated or unsaturated linear or C3-C100 cyclic hydrocarbon, C1 -C100 saturated or unsaturated linear or C3-C100 cyclic hydrocarbon having at least one heteroatom, C1- C100 saturated or unsaturated linear or C3-C100 cyclic hydrocarbon having at least one heteroatom and at least one D, aromatic group, substituted aromatic group, or a combination thereof;R9 is selected from H, D, C1 -C100 saturated or unsaturated linear or C3-C100 cyclic hydrocarbon, C1 -C100 saturated or unsaturated linear or C3-C100 cyclic hydrocarbon having at least one heteroatom, C1-C100 saturated or unsaturated linear or C3-C100 cyclic hydrocarbon having at least one heteroatom and at least one D, OH, NH2, Cl, F, I and Br;R10 is selected from H, Cl, F, I, Br and methoxy;R11 is OH, OR12, SH, SR13, NH2, NHD, ND2, NRuR’u, wherein R12 is an alcohol protecting group, R13 is a thiol protecting group, and Ru and R’14 each is independently H, D, or an amine protecting group; the heteroatom is selected from N, O, S, P, Cl, Br, I and a combination thereof; and X is O, S, or NH.

[0124] Any pharmaceutically suitable carrier disclosed herein can be suitable.

[0125] In some cases, v is in a range of from 1 -100, 1-90, 1 -80, 1-70, 1-60, 1-50, 1-40, 1-30, 1-20, 1-18, 1-16, 1-14, 1-12, 1-10, 1-8, 1 -6, 1-4, 1-3, or 1 -2 and w is in a range of from 1 - 1000. In some cases, v is 1. In some cases, w is in a range of from 1-1000, 1- 900, 1-800, 1-700, 1 -600, 1-500, 1-400, 1-300, 1-200, 1-150, 1-120, 1-100, 1-80, 1-70, 1- 60, 1-50, 1-40, 1-30, 1-20, 1-18, 1-16, 1-14, 1-12, 1-10, 1-8, 1-6, 1-4, 1-2, and v is in arrange of from 1-100. In some cases, v is 1-200. In some cases, w is in a range of from 1 -1000, 1-900, 1-800, 1-700, 1-600, 1-500, 1-400, 1-300, 1-200, 1-180, 1-160, 1-140, 1- 120, 1-100, 1-80, 1-60, 1-40, 1-30, 1-20, 1-10, or 1-2 and v is in a range of from 1 - 100. In some cases, w is 1. In some cases, v and w are in a combination of the ranges disclosed above.

[0126] In some cases, X is O. In some cases, X is S. In some cases, X is NH.

[0127] In some cases, R, R’ and R” each is H, C1 -22 alkyl group, C1-20 alkyl group, CI- 18 alkyl group, C1-16 alkyl group, C1-14 alkyl group, C1-12 alkyl group, C1-10 alkyl group, C1-8 alkyl group, C1-6 alkyl group, aromatic group, or substituted aromatic group. In some cases, at least one of the R, R’ and R” is C1-22 alkyl group, C1-20 alkyl group, C1-18 alkyl group, C1 -16 alkyl group, C1-14 alkyl group, C1 -12 alkyl group, C1-10 alkyl group, C1-8 alkyl group, or C1-6 alkyl group. In some cases, at least one of the R, R’ and R” is C22 alkyl group, C20 alkyl group, C18 alkyl group, C16 alkyl group, C14 alkyl group, C12 alkyl group, C10 alkyl group, or C8 alkyl group, providing hydrophobicity to the modified exatecan.

[0128] In some cases, at least one of the R, R’ and R” is C1-22 alkyl group.

[0129] In some cases, v=1 , w=1 , and R is selected from methyl, ethyl, benzyl, CNCH2-, cyclopropyl, benzene, 1 ,1 ,1 -trifluoroethyl, n-propyl, and a combination thereof. In some cases, R, R’, R”, R3, R4, R3’, R4’, Rs, Re, R7, Rs and R15 each is independently H, D, CI- 022 saturated or unsaturated hydrocarbon, C1-C22 saturated or unsaturated hydrocarbon having at least one D, 01-022 saturated or unsaturated hydrocarbon having at least one heteroatom, C1 -C22 saturated or unsaturated hydrocarbon having at least one heteroatom and at least one D, aromatic group, substituted aromatic group, or a combination thereof. In some cases, R3, R4, R3’, R4’, Rs, Re, R7, Rs and R15 each is independently H or D. In some cases, R9 is a methyl. In some cases, R10 is F. In some cases, R11 is OH. In some cases, the PBA of Formula ID 1 , 2 or 3 can have a combination of the groups disclosed hereabove.

[0130] Any of the alcohol protecting groups, thiol protecting groups, or amine protecting groups disclosed above can be suitable.

[0131] In some cases, the PBA can have a formula selected from Formula ID 1001-1135, a pharmaceutically acceptable salt thereof, a solvate thereof, a prodrug thereof, an isomer thereof, an isotope derivative thereof, and a combination thereof.

[0132] In some cases, the PBA can comprise a formula selected from Formula ID 1 , 1-1 , 1-2, 2, 2-1 , 2-2, 3, 3-1 , 3-2, 1001 , 1002, 1003, 1004, 1005, 1006, 1007, 1008, 1009, 1010, 1011 , 1012, 1013, 1014, 1015, 1016, 1017, 1018, 1019, 1020, 1021 , 1022, 1023, 1024,1025, 1026, 1027, 1028, 1029, 1030, 1031 , 1032, 1033, 1034, 1035, 1036, 1037, 1038,1039, 1040, 1041 , 1042, 1043, 1044, 1045, 1046, 1047, 1048, 1049, 1050, 1051 , 1052,1053, 1054, 1055, 1056, 1057, 1058, 1059, 1060, 1061 , 1062, 1063, 1064, 1065, 1066,1067, 1068, 1069, 1070, 1071 , 1072, 1073, 1074, 1075, 1076, 1077, 1078, 1079, 1080,1081 , 1082, 1083, 1084, 1085, 1086, 1087, 1088, 1089, 1090, 1091 , 1092, 1093, 1094,1095, 1096, 1097, 1098, 1099, 1100, 1101 , 1102, 1103, 1104, 1105, 1 106, 1107, 1108,1109, 1 110, 1111 , 1112, 1 113, 1114, 1115, 1116, 1117, 1118, 11 19, 1 120, 1121 , 1122, 1123, 1124, 1125, 1126, 1127, 1128, 1129, 1130, 1131 , 1132, 1133, 1134, 1135, the additional cytotoxic agent comprising calicheamicin, DM1 , DM4 (Ravtansine), Dxd (deruxtecan), exatecan, IRDye700DX, MMAE (Monomethyl auristatin E), vedotin, MMAF (Monomethyl auristatin F), Pseudomonas exotoxin A, pyrrolobenzodiazepine (PBD), SG3199 (PBD dimer), SN-38, irinotecan, camptothecin, amanitin, duocarmycins (DNA- alkylators), halichondrin, Eribulin (Halaven®), norhalichondrin B, maytansinoids, maitansine (INN), maytansine, PUN-159682, tubulysins, tubulysin, pretubulysin (PT), tubulysin A (TubA), tubulysin M, a pharmaceutically acceptable salt thereof, a solvate thereof, a prodrug thereof, an isomer thereof, an isotope derivative thereof, or a combination thereof.

[0133] In some cases, the pharmaceutical composition can comprise a PBA having a formula selected from Formula ID 1 , 1-1 , 1-2, 2, 2-1 , 2-2, 3, 3-1 , 3-2, and 1001-1135, the additional cytotoxic agent comprising calicheamicin, DM1 , DM4 (Ravtansine), Dxd (deruxtecan), exatecan, IRDye700DX, MMAE (Monomethyl auristatin E), vedotin, MMAF (Monomethyl auristatin F), Pseudomonas exotoxin A, pyrrolobenzodiazepine (PBD), SG3199 (PBD dimer), SN-38, irinotecan, camptothecin, amanitin, duocarmycins (DNA- alkylators), halichondrin, Eribulin (Halaven®), norhalichondrin B, maytansinoids, maitansine (INN), maytansine, PUN-159682, tubulysins, tubulysin, pretubulysin (PT), tubulysin A (TubA), tubulysin M, a pharmaceutically acceptable salt thereof, a solvate thereof, a prodrug thereof, an isomer thereof, an isotope derivative thereof, or a combination thereof.

[0134] In some cases, the pharmaceutical composition can further comprise: a targeting bioactive agent (TBA) comprising binding affinity to a target molecule or a target cell; and wherein the PBA is reacted and covalently linked to the TBA directly or indirectly.

[0135] In some cases, the pharmaceutical composition can further comprise: a TBA comprising binding affinity to a target molecule that comprises a tumor antigen (Ag), a coupler, a polymer a linker, or a combination thereof, wherein the TBA can be covalently linked to the PBA directly or indirectly.

[0136] In some cases, the pharmaceutical composition can comprise the PBA covalently linked to the TBA directly.

[0137] In some cases, the pharmaceutical composition can comprise a PBA covalently linked to the TBA directly (Formula ID 30), via a polymer (Formula ID 31 ), via a polymerand one or more linkers (Formula ID 32, 33, and 34), via a coupler (Formula ID 35), via a coupler, a polymer and one or more linkers (Formula ID 36, 37, and 38):wherein the PBAs can be selected from the Formula ID 1 , 1-1 , 1-2, 2, 2-1 , 2-2, 3, 3-1 , 3-2, and 1001-1135, a pharmaceutically acceptable salt thereof, a solvate thereof, a prodrug thereof, an isomer thereof, an isotope derivative thereof, a reacted PBA thereof, and a combination thereof; and n is an integer and can be in a range of from 1 to 100.

[0138] In some cases, n can be in a range of from 1 to 24, 1 to 20, 1 to 18, 1 to 16, 1 to 14, 1 to 12, 1 to 10, 1 to 8, 1 to 6, 1 to 4, or 1 to 2.

[0139] The linker, the Linker L1 and L2 each can be a cleavable linker and can be the same or different. In some cases, the linker, the Linker L1 and L2 each can be indecently a same or different cleavable linker. In some cases, the linker, the Linker L1 and L2 all can be indecently same or different cleavable linkers.

[0140] The linker can comprise one or more cleavable linkers. In some cases, the cleavable linker can be selected from the Formula ID 101-141 , or a combination thereof.

[0141] In some cases, the pharmaceutical composition can comprise a formula selected from Formula ID 30, 31 , 32, 33, 34, 35, 36, 37, 38, 39, 40 or a combination thereof; wherein, the TBA can be selected from a peptide; a protein; DNA; RNA; nucleic acids; oligo nucleic acids; mRNA; siRNA; sgRNA; an antibody selected from a monoclonal antibody, a fragment of a monoclonal antibody, a polyclonal antibody, a fragment of a polyclonal antibody, a synthetic antibody, a fragment of a synthetic antibody, antigen-bindingportions thereof, or a combination thereof; an agonist; an activator; an inhibitor; an antagonist; a ligand of cell surface receptor; a naked nucleic acid; a nucleic acid; an antisense nucleic acid, anti-CD19; a ribozyme; a virus; a virus-like particles and a combination thereof; chimeric antigen receptor or chimeric antigen T-cell receptor (CAR- T), cytokines; granulocyte colony stimulating factor (G-CSF); interferon a; interferon a2a; interleukins; lectins; Neupogen (Filgrastim); siRNAs; T-cell receptor (TCR); a member of a small molecule binding pair, and a combination thereof; the coupler, when present, can be selected from Formula ID 201 , 202, 203, 204, 205, 026, 207, 208, 209, and a combination thereof; the polymer can be selected from a peptide, a polyethylene glycol (PEG), polyoxazoline (POX), or a combination thereof, wherein the polyoxazoline (POX) comprises a linear portion, a branched portion, or a combination thereof, and wherein the polyoxazoline (POX) comprises poly(2-methyloxazoline), poly(2-ethyloxazoline), poly(2- propyloxazoline), poly(2-isopropyloxazoline), poly(2-substituted oxazoline), and a combination thereof; and the PBA can comprise a formula selected from Formula ID 1 , 1-1 , 1-2, 2, 2-1 , 2-2, 3, 3-1 , 3-2, 1001 , 1002, 1003, 1004, 1005, 1006, 1007, 1008, 1009, 1010, 1011 , 1012, 1013, 1014, 1015, 1016, 1017, 1018, 1019, 1020, 1021 , 1022, 1023, 1024, 1025, 1026, 1027,1028, 1029, 1030, 1031 , 1032, 1033, 1034, 1035, 1036, 1037, 1038, 1039, 1040, 1041 ,1042, 1043, 1044, 1045, 1046, 1047, 1048, 1049, 1050, 1051 , 1052, 1053, 1054, 1055,1056, 1057, 1058, 1059, 1060, 1061 , 1062, 1063, 1064, 1065, 1066, 1067, 1068, 1069,1070, 1071 , 1072, 1073, 1074, 1075, 1076, 1077, 1078, 1079, 1080, 1081 , 1082, 1083,1084, 1085, 1086, 1087, 1088, 1089, 1090, 1091 , 1092, 1093, 1094, 1095, 1096, 1097,1098, 1099, 1100, 1101 , 1 102, 1103, 1104, 1105, 1106, 1107, 1108, 1 109, 1110, 1111 ,1112, 1 113, 1114, 1115, 1 116, 1117, 1118, 1119, 1120, 1121 , 1122, 1 123, 1124, 1125,1 126, 1 127, 1128, 1129, 1 130, 1 131 , 1132, 1133, 1 134, 1135, the additional cytotoxic agent comprising calicheamicin, DM1 , DM4 (Ravtansine), Dxd (deruxtecan), exatecan, IRDye700DX, MMAE (Monomethyl auristatin E), vedotin, MMAF (Monomethyl auristatin F), Pseudomonas exotoxin A, pyrrolobenzodiazepine (PBD), SG3199 (PBD dimer), SN- 38, irinotecan, camptothecin, amanitin, duocarmycins (DNA-alkylators), halichondrin, Eribulin (Halaven®), norhalichondrin B, maytansinoids, maitansine (INN), maytansine, PUN-159682, tubulysins, tubulysin, pretubulysin (PT), tubulysin A (TubA), tubulysin M, a pharmaceutically acceptable salt thereof, a solvate thereof, a prodrug thereof, an isomer thereof, an isotope derivative thereof, or a combination thereof.

[0142] In some cases, polymer can comprise a single amino acid residue, dipeptide, tripeptide, or polypeptide. In some cases, as mentioned above, the polymer can comprise a peptide comprising in a range of from 1 to 100 amino acid residues. In some cases, a polymer can comprise a lysine, glutamic acid, glutamine, arginine, valine, or a combination thereof.

[0143] In some cases, the polymer, the PBA, or the linker can be attached to the antibody, also referred to as “conjugated to the antibody” or “bioconjugation”, via S-bond (such as thiol-bond or thiol conjugation, via cysteine conjugation), N-bond (such as via lysine conjugation), glycol bond, modified glycol bond, GIcNAc (N-acetylglucosamine), substituted GIcNAc, GIcNAc-galactose, GIcNAc-mannose, GIcNAc-glucose, GIcNAc- glucuronic acid, GIcNAc-fucose, GIcNAc-N-acetylneuraminic acid, site specific bond, random bond, or any other bonds or conjugations known to or developed by those skilled in the art. In some cases, the couplers disclosed herein can be suitable. The S-bond conjugation can include the aforementioned cystine-based conjugation, such as reactions via maleimide, for example, with a MC linker, a MC-VC-PABC linker, MC-VC-PAB-NH2 linker, a MC-GGFG-AM, a CL2A linker, and maleimide-VA-PABC linker; alkynyl carboxylic acid derivatives; 5-methylene pyrrolone (5MP); 5,5’-dithiobis-(2-nitrobenzoic acid) (DTNB); phenyloxadiazole sulfone (PODS); cyclooctyne, an alkyne moiety in aza- dibenzocyclooctyne (DBCO) reagent that is shown to react with a cysteine residue as a side reaction; via 3-ary Ipropionitrile (APN); perfluoroarene; ethynylbenziodoxolone (EBX); bicyclo[1.1.0]butane (BOB) carboxylic amide; allenamide; and disulfide functionalization such as with a bis-reactive linker that undergoes reactions with both thiol residues (also known as disulfide rebridging) to produce an ADC having a controlled DAR when modifying antibodies with four reducible interchain disulfides. In some cases, the polymer, the PBA, or the linker can be attached to the antibody via one or more couplers selected from Formula ID 201 , 202, 203, 204, 205, 026, 207, 208, 209, and a combination thereof.

[0144] In some cases, bioconjugation to other amino acid residues of the antibody different from or in addition to cysteine (Cys), such as lysine (Lys), tyrosine (Tyr), tryptophan (Trp), methionine (Met), other natural or un-natural amino acids, can also be suitable. Lysine conjugation can include N-hydroxysuccinimide (NHS) ester, isocyanates and isothiocyanates, 4-azidobenzoyl fluoride (ABF), p-lactams, Phospha-Mannich, a,p- unsaturated sulfonamide, sulfonyl acrylate, and others. Tyrosine conjugation can include Mannich-type three component reactions, diazonium salt reactions, 4-phenyl-3H-1 ,2,4- triazole-3,5(4H)-dione (PTAD), phenothiazine, and other reactions. Tryptophan conjugation can include 9-azabicyclo [3.3.1]nonane-3-one-N-oxyl (keto-ABNO), N-substituted pyridinium salts, and other reactions. Methionine conjugation can include oxaziridine, hypervalent iodine, lumiflavin-catalysis, and others. In some cases, conjugation methods, such as the click chemistry, proximity-induced site-specific antibody conjugation chemistry (pCIick), or a combination thereof, can be suitable,

[0145] In some cases, the pharmaceutical composition can comprise a Nano-ADC that can comprise bio-conjugated antibody, polymer, PBA, and linker via bioconjugation to one or more amino acids residues of the antibody selected from lysine (Lys), tyrosine (Tyr), tryptophan (Trp), methionine (Met), one or more genetically engineered non-natural amino acids (NNAA), or a combination thereof.

[0146] In some cases, antibodies having genetically engineered non-natural amino acids (NNAA), such as p-acetylphenylalanine and p-azidomethylphenyl alanine, can also be suitable for producing antibody-drug conjugates. In some cases, the pharmaceutical composition can comprise a Nano-ADC that can comprise an antibody having genetically engineered non-natural amino acids (NNAA), such as p-acetylphenylalanine and p- azidomethylphenyl alanine, enzymatic conjugations such as those produced with bacterial transglutaminase (TGase), Sortase, Glycosyl transferase, Formylglycine- generating enzyme (FGE), or Isoprenoid transferase, or a combination thereof.

[0147] In some cases, a coupler can be linked to a polymer, linker and payload (PBA), and then conjugated to an antibody via one of the conjugation methods described herein. In some cases, the pharmaceutical composition can comprise a Nano-ADC produced by the conjugation methods disclosed herein.

[0148] Alternatively, in some cases, a coupler can be first linked to an antibody to produce an activated antibody via reductive coupling and then linked to a PBA, directly or via a polymer, a linker or a combination thereof. The conjugated products can be purified as described herein. In some cases, the pharmaceutical composition can comprise a Nano-ADC produced by first linking a coupler to an antibody and then linking a thiolated PBA. In some cases, the pharmaceutical composition can comprise a Nano-ADC produced by first linking a coupler to an antibody and then linking thiolated one or more polymers, one or more linkers and one or more PBAs. In some cases, the pharmaceutical composition can comprise a Nano-ADC produced by first linking a coupler to an antibody and then linking thiolated one or more polymers, one or more polymer side chains, one or more linkers and one or more PBAs. Any cysteine-based couplers disclosed herein or known to those skilled in the art can be suitable. In some cases, the coupler can comprise succinimidyl-4-(N-maleimidomethyl)cyclohexane-1 -carboxylate (SMCC), maleimide (Mai), dibromomaleimide (DBM), maleimidocaproyl (MC), maleimidomethyl cyclohexane-1 -carboxylate (MCC), Mal-PEG, or a combination thereof. In some cases, the polymer, linker or PBA can be thiolated.

[0149] In some cases, the linker can comprise one or more cleavable linkers and can be selected from Formula ID 101-141 , and a combination thereof.

[0150] In some cases, the pharmaceutical composition can comprise at least two PBAs that can be the same or different. In some cases, the pharmaceutical composition can comprise at least two different PBAs covalently linked to each polymer, such as each PEOX polymer or each PEG polymer, wherein each of the at least two different PBAs can be selected from at least two different natural or synthetic small molecule-based drugs, inorganic-based drugs, toxin molecules having cytotoxicity, molecules having cytotoxicity to target cells, derivatives thereof, or a combination thereof. Each of the PBAs can be independently selected from the PBAs disclosed herein including the Formula ID 1 , 1-1 , 1-2, 2, 2-1 , 2-2, 3, 3-1 , 3-2, and 1001-1135, a pharmaceutically acceptable salt thereof, a solvate thereof, a prodrug thereof, an isomer thereof, an isotope derivative thereof, a reacted PBA thereof, and a combination thereof.

[0151] In some cases, the PBA can further comprise calicheamicin, DM1 (antimicrotubule agent T-DM1 , a derivative of maytansine), DM4 (Ravtansine), Dxd (deruxtecan), exatecan, IRDye700DX (a near-infrared (NIR) photosensitizer suitable for near infrared (NIR) photoimmunotherapy (PIT)), MMAE (Monomethyl auristatin E, also known as vedotin), MMAF (Monomethyl auristatin F), pseudomonas exotoxin A (a potent exotoxin produced by the bacterium Pseudomonas aeruginosa), pyrrolobenzodiazepine (PBD), SG3199 (a cytotoxic DNA minor groove interstrand crosslinking pyrrolobenzodiazepine (PBD) dimer), SN-38, irinotecan, camptothecin, amanitin (an inhibitor of RNA Polymerase II), duocarmycins (DNA-alkylators), halichondrin (Eribulin, also known Halaven®, under respective trademark), norhalichondrin B, maytansinoids, maitansine (INN), maytansine, PUN-159682 (an inhibitor of DNA Topoisomerase II), tubulysins, tubulysin, pretubulysin (PT), tubulysin A (TubA), tubulysin M, or a combination thereof.

[0152] In some cases, the pharmaceutical composition can comprise at least one polymer side chain attached to the polymer. In some cases, the polymer side chain can be free from any PBAs. In some cases, the pharmaceutical composition can comprise one or more polymer side chains that at least one of the side chains can be attached to one or more PBAs. In some cases, the pharmaceutical composition can comprise one or more polymer side chains that each of the side chains can be attached to one or more PBAs.

[0153] In some cases, the polymer can comprise polyoxazoline (POX) having a branched portion that can comprise two or more branches, and wherein at least one of the two ormore branches can be the polymer side chain and can be free from the PBA. In some cases, each of the two or more branches can be the polymer side chain and can be attached to one or more PBAs.

[0154] In some cases, the pharmaceutical composition can further comprise a polymer side chain that is covalently linked to the TBA, the polymer, or a combination thereof, wherein the polymer side chain can comprise POX comprising poly(2-methyloxazoline), poly(2-ethyloxazoline), poly(2-propyloxazoline), poly(isopropyloxazoline), or a combination thereof.

[0155] In some cases, the pharmaceutical composition can comprise the polymer that comprises a peptide, polyoxazoline (POX), wherein the polyoxazoline (POX) can comprise a linear portion, a branched portion, or a combination thereof, and wherein the polyoxazoline (POX) comprises poly(2-methyloxazoline), poly(2-ethyloxazoline), poly(2- propyloxazoline), poly(2-isopropyloxazoline), poly(2-substituted oxazoline), and a combination thereof, polyoxazoline (POX) having a branched portion that can comprise two or more branches, and one or more PBAs selected from the PBA having the Formula ID 1 , 1-1 , 1-2, 2, 2-1 , 2-2, 3, 3-1 , 3-2, and 1001-1135, the additional cytotoxic agent comprising calicheamicin, DM1 (antimicrotubule agent T-DM1 , a derivative of maytansine), DM4 (Ravtansine), Dxd (deruxtecan), exatecan, IRDye700DX (a nearinfrared (NIR) photosensitizer suitable for near infrared (NIR) photoimmunotherapy (PIT)), MMAE (Monomethyl auristatin E, also known as vedotin), MMAF (Monomethyl auristatin F), pseudomonas exotoxin A (a potent exotoxin produced by the bacterium Pseudomonas aeruginosa), pyrrolobenzodiazepine (PBD), SG3199 (a cytotoxic DNA minor groove interstrand crosslinking pyrrolobenzodiazepine (PBD) dimer), SN-38, irinotecan, camptothecin, amanitin (an inhibitor of RNA Polymerase II), duocarmycins (DNA- alkylators), halichondrin (Eribulin, also known Halaven®, under respective trademark), norhalichondrin B, maytansinoids, maitansine (INN), maytansine, PUN-159682 (an inhibitor of DNA Topoisomerase II), tubulysins, tubulysin, pretubulysin (PT), tubulysin A (TubA), tubulysin M, a pharmaceutically acceptable salt thereof, a solvate thereof, a prodrug thereof, an isomer thereof, an isotope derivative thereof, and a combination thereof.

[0156] In some cases, polymer can comprise a single amino acid residue, dipeptide, tripeptide, or polypeptide. In some cases, as mentioned above, the polymer can comprise a peptide comprising in a range of from 1 to 100 amino acid residues. In some cases, a polymer can comprise a lysine, glutamic acid, glutamine, arginine, valine, or a combination thereof.

[0157] The term “reacted compound,” “reacted TBA,” “reacted PBA,” or “reacted mExa” means a compound having one or more functional groups reacted with another functional group or another molecule. In some cases, the -N(CH2)2-OH, -S(CH2)2-OH, or -O(CH2)2- OH terminal group in each of the modified exatecan Formula ID 1 , 1-1 , 1-2, 2, 2-1 , 2-2, 3, 3-1 and 3-2, is reacted to another functional group to be covalently linked to the TBA directly or indirectly. In some cases, the -N(CH2)2-OH terminal group in each of the modified exatecan Formulas (1 )-(3) is reacted to another functional group to be covalently linked to the TBA directly or indirectly to form an ADC. In some cases, the -N(CH2)2-OH, -S(CH2)2-OH, or -O(CH2)2-OH terminal group in each of the modified exatecan Formulas (1 )-(3) is reacted to a linker. In some cases, the modified exatecan selected from Formula ID, 1 , 1-1 , 1-2, 2, 2-1 , 2-2, 3, 3-1 , 3-2, and 1001-1135 each is reacted to a linker, a polymer, a coupler, or a combination thereof, to form an antibody drug conjugate (ADC), i.e. , Nano-ADC. In some cases, a Nano-ADC can have a formula comprising Formula ID 3001-3074, or a combination thereof.

[0158] Each of the ranges is intended as a continuous range including every value between the minimum and maximum values and including the minimum and maximum cited values.

[0159] The pharmaceutical composition can comprise the PBA disclosed herein that is reacted, i.e., covalently linked to a polymer a linker, or a combination thereof. In some cases, the linker can be a cleavable linker. In some cases, the PBA, the polymer and the cleavable linker can be positioned as described hereafter and illustrated in FIG. 7A-FIG. 7H.

[0160] In some cases, the pharmaceutical composition can comprise a reacted PBA having a formula selected from Formula ID 1 , 1-1 , 1-2, 2, 2-1 , 2-2, 3, 3-1 , 3-2, and 1001- 1135, a pharmaceutically acceptable salt thereof, a solvate thereof, a prodrug thereof, an isomer thereof, an isotope derivative thereof, and a combination thereof.

[0161] In some cases, the TBA can comprise an antibody, a fragment thereof, a bispecific antibody thereof, a tri-specific antibody thereof, a tetra-specific antibody thereof, an antigen-binding portion thereof, or a combination thereof, herein collectively referred to as “antibody” or “Ab”.

[0162] In some cases, unlimiting examples of schematic illustrations of the pharmaceutical composition of this disclosure can be shown in FIG. 1A-FIG. 1 D. The TBA (1) can be an antibody that can be linked to a PBA (3) via a linker (2). The TBA- linker-PBA can form nanoaggregates (5) (FIG. 1A). The pharmaceutical composition can comprise a cleavable linker (8) forming nanoaggregates (6) having the cleavable linker.The pharmaceutical composition can further comprise a polymer (4), two cleavable linkers (8a and 8b) forming nanoaggregates (7) having the cleavable linkers and the polymer (FIG. 1C). The nanoaggregates can further form nanoaggregates complex (9) that can comprise 2 or more aggregates (FIG. 1 D). The nanoaggregates and nanoaggregates complex can be collectively referred to as Nano-ADC aggregates when the pharmaceutical composition is a Nano-ADC.

[0163] In some cases, further schematic illustrations of some representative examples of the pharmaceutical composition having a polymer (4a) covalently linked to a TBA (1) and a PBA, such as a first PBA (3) and a second PBA (3’) are shown in FIG. 2A-FIG. 2E. In some cases, the pharmaceutical composition can comprise a Linker L1 (2a) and a Linker L2 (2b) without any cleavable linkers (FIG. 2A). Each TBA can be covalently linked to one or more PBAs with n as disclosed herein. In some cases, the pharmaceutical composition can comprise at least a first cleavable linker (8a) in the Linker L1 (2a) between the TBA (1) and the polymer (4a) (FIG. 2B). In some cases, the pharmaceutical composition can comprise at least one second cleavable linker (8b) in the Linker L2 between the first PBA (3) and the polymer (4a) (FIG. 2C). In some cases, the pharmaceutical composition can comprise at least one first cleavable linker (8a) in the Linker L1 between the TBA (1) and the polymer (4a) and at least one second cleavable linker (8b) in the Linker L2 between the first PBA (3) and the polymer (4a) (FIG. 2D). In some cases, the pharmaceutical composition can comprise multiple same or different polymers, such as the polymer (4a) and a subsequent polymer (4a’) and same or different PBAs, such as a first PBA (3) and a second PBA (3’), a subsequent first cleavable linker (8a’) and a subsequent second cleavable linker (8b’) (FIG. 2E). The (R1 ) (or simply shown as R1 ) (10) and (R2) (or simply shown as R2) (11 ) represent the reacted first terminal group and the reacted second terminal group, respectively. The first and second PBAs can be the same or different. The polymer (4a) and the subsequent polymer (4a’) can be the same or different.

[0164] As used herein throughout this disclosure, the polymers (4a, 4a’, 4b, 4c) can be a linear polymer, asymmetric branched polymer, symmetric branched polymer, or a combination thereof. Legends: Open rectangle, Linker L1 ; Hatched rectangle, Linker L2; Solid rectangle, the polymer of this disclosure that can be a linear polymer, asymmetric branched polymer, symmetric branched polymer, or a combination thereof; Crossed circle, cleavable bond; Solid circle, PBA. Throughout this disclosure, each TBA can be covalently linked to one or more PBAs, n is an integer and can be in a range of from 1 to 100.

[0165] In some cases, examples of the pharmaceutical composition comprising a polymer having asymmetrical branches are shown in FIG. 3A-FIG. 3D. In some cases, a pharmaceutical composition can comprise a TBA (1) covalently linked to a polymer having asymmetrical branches (4b) that is covalently linked to at least one PBA (3) (FIG. 3A) without any cleavable linkers. In some cases, a pharmaceutical composition can comprise a TBA (1) covalently linked to a polymer having asymmetrical branches (4b) that is covalently linked to at least one PBA (3) and at least first cleavable linker (8a) between the TBA (1) and polymer (4b) (FIG. 3B). In some cases, a pharmaceutical composition can comprise a TBA (1 ) covalently linked to a polymer having asymmetrical branches (4b) that is covalently linked to at least one PBA (3) and at least one second cleavable linker (8b) between the polymer (4b) and each of the PBAs (3) (FIG. 3C). In some cases, a pharmaceutical composition can comprise a TBA (1) covalently linked to a polymer having asymmetrical branches (4b) that is covalently linked to at least one PBA (3), at least one first cleavable linker (8a) between the TBA (1) and polymer (4b) and at least one second cleavable linker (8b) between the polymer (4b) and at least one of the PBA (3) (FIG. 3D). In some cases, a pharmaceutical composition can comprise a TBA (1) covalently linked to a polymer having asymmetrical branches (4b) that is covalently linked to at least one PBA (3), at least one first cleavable linker (8a) between the TBA (1 ) and polymer (4b) and at least one second cleavable linker (8b) between the polymer (4b) and each of the PBAs (3).

[0166] Some examples of the pharmaceutical composition comprising a polymer having symmetrical branches are shown in FIG. 4A - FIG. 4D. In some cases, the pharmaceutical composition can comprise a TBA (1 ) covalently linked to a polymer having symmetrical branches (4c) that is covalently linked to at least one PBA (3) (FIG. 4A) without any cleavable linker. In some cases, the pharmaceutical composition can comprise a TBA (1) covalently linked to a polymer having symmetrical branches (4c) that is covalently linked to at least one PBA (3) and at least one first cleavable linker (8a) between the TBA (1) and polymer (4c) (FIG. 4B). In some cases, the pharmaceutical composition can comprise a TBA (1) covalently linked to a polymer having symmetrical branches (4c) that is covalently linked to at least one PBA (3) and at least one second cleavable linker (8b) between the polymer (4c) and at least one of the PBAs (3) or each of the PBAs (FIG. 4C). In some cases, the pharmaceutical composition can comprise a TBA (1 ) covalently linked to a polymer having symmetrical branches (4c) that is covalently linked to at least one PBA (3), at least one first cleavable linker (8a) between the TBA (1)and polymer (4c) and at least one second cleavable linker (8b) between the polymer (4c) and at least one of the PBAs (3) or each of the PBAs (FIG. 4D).

[0167] I n some cases, the pharmaceutical composition can comprise one cleavable linker between the TBA and the polymer. Once the pharmaceutical composition is subject to a proper cleavage condition, such as reducing agent for a disulfide bond linker, an enzyme for a peptide linker, or other conditions, the cleavable linker can be cleaved resulting in the dissociation of the TBA from the pharmaceutical composition and polymer-PBA complex. The polymer-PBA can form polymer-PBA aggregates in aqueous environments such as in blood stream, in plasma, or in cytosol (FIG. 5). For some PBAs, albumin in the blood may also interact with the PBA forming nanoaggregates that comprise the polymer, the PBA and albumin, wherein the polymer and the PBA are linked covalently forming individual nanoaggregates (12) or nanoaggregates complex (12a).

[0168] I n some cases, the pharmaceutical composition can comprise one cleavable linker between the TBA and the polymer and at least one cleavable linker between the polymer and the PBA or at least one of the PBAs when multiple PBAs are present. Once the pharmaceutical composition is subject to a proper cleavage condition in vivo or in vitro, the cleavable linker can be cleaved resulting in the dissociation of the TBA from the pharmaceutical composition, polymer and PBA. The dissociated polymer and PBA can form polymer-PBA aggregates in aqueous environments such as in blood stream, in plasma, or in cytosol (FIG. 6A). For some PBAs, albumin in the blood may also interact with the PBA forming nanoaggregates that comprise the polymer, the PBA and albumin, wherein the polymer and the PBA are aggregated non-covalently forming individual nanoaggregates (13) or nanoaggregates complex (13a). The dissociated polymer and PBA can form polymer-PBA aggregates that are free from the TBA.

[0169] In some cases, the PBA can comprise the modified exatecan (mExa), one linker (2a) between a TBA) (1) and a polymer (4) and at least one second cleavable linker (8b) between the polymer and the modified exatecan (3a) (FIG. 6B). In some cases, the mExa can be selected from Formula ID 1 , 1-1 , 1-2, 2, 2-1 , 2-2, 3, 3-1 , 3-2, and 1001-1135 a pharmaceutically acceptable salt thereof, a solvate thereof, a prodrug thereof, an isomer thereof, an isotope derivative thereof, and a combination thereof. Once the pharmaceutical composition is subject to a proper cleavage condition in vivo or in vitro, the cleavable linker can be cleaved resulting in the dissociation of the TBA from the pharmaceutical composition, polymer and mExa. The dissociated mExa (3a) can form nanoaggregates in aqueous environments such as in blood stream, in plasma, or in cytosol (FIG. 6B), such as with albumin. The poly(2-ethyloxazoline) (PEOX) or a PEOXderivative modified exatecan as disclosed in Formula ID 1 , 1-1 , 1-2, 2, 2-1 , 2-2, 3, 3-1 , 3- 2, and 1001-1135 can form individual nanoaggregates (13’) or nanoaggregates complex (13’a). The PEOX ora PEOX derivative modified exatecan can form nanoaggregates that are free from the TBA, such as an antibody.

[0170] In some cases, the PBA can comprise the modified exatecan (mExa) (3a), a first cleavable linker (8a) between a TBA (1) and a polymer (4) and, optionally, a Linker L2 (2b) between the polymer and the modified exatecan (3a) (FIG. 6C), wherein the Linker L2 can be non-cleavable. Once the pharmaceutical composition is subject to a proper cleavage condition in vivo or in vitro, the cleavable linker can be cleaved resulting in the dissociation of the TBA from the pharmaceutical composition, polymer and mExa. The dissociated mExa (3a) together with the polymer and the optional Linker L2 can form nanoaggregates in aqueous environments such as in blood stream, in plasma, or in cytosol (FIG. 6C). The poly(2-ethyloxazoline) (PEOX) or a PEOX derivative modified exatecan as disclosed in Formula ID 1 , 1-1 , 1-2, 2, 2-1 , 2-2, 3, 3-1 , 3-2, 1001-1135, a pharmaceutically acceptable salt thereof, a solvate thereof, a prodrug thereof, an isomer thereof, an isotope derivative thereof, and a combination thereof, and the polymer, and optionally the albumin, can form individual nanoaggregates (13”) or nanoaggregates complex (13”a). The PEOX or a PEOX derivative modified exatecan can form nanoaggregates that are free from the TBA, such as an antibody.

[0171] In some cases, a pharmaceutical composition can comprise an antibody-drug conjugate (ADC), wherein the TBA can be an antibody, such as an anti-HER2 antibody. In some cases, an antibody-drug conjugate of this disclosure (herein “Nano-ADC”) can comprise an antibody, a polyethyleneglycol (PEG) with 1-100 monomer units (m=1-100), a cleavable linker, such as a linker GGFG, a linker VAPAB, ora combination thereof, and a PBA. Each Nano-ADC can comprise in a range of from 1 -100 of the PBA, i.e., n is an integer in a range of from 1 to 100.

[0172] Some schematic examples of ADCs are shown in FIG. 7A - FIG. 7H. In some cases, an ADC can comprise an Ab-Polymer-Linker-PBA structure without a cleavable linker (FIG. 7A). In some cases, an ADC can comprise an Ab-Linker-Polymer-PBA structure without a cleavable linker (FIG. 7B). In some cases, an ADC can comprise an Ab-PEG-GGFG-mExa structure having a GGFG cleavable linker between the polymer, such as a PEG, and the mExa (FIG. 7C). In some cases, an ADC can comprise an Ab- GGFG-PEG-mExa structure having a GGFG cleavable linker between the antibody and the polymer, such as a PEG (FIG. 7D). In some cases, an ADC can comprise an Ab-PEG- VCPAB-mExa structure having a VCPAB cleavable linker between the polymer, such aPEG, and the mExa (FIG. 7E). In some cases, an ADC can comprise an Ab-VCPAB- PEG-mExa structure having a VCPAB cleavable linker between the antibody and the polymer, such as a PEG (FIG. 7F). In some cases, an ADC can comprise an Ab-PEG- VAPAB-mExa structure having a VAPAB cleavable linker between the polymer, such as a PEG, and the mExa (FIG. 7G). In some cases, an ADC can comprise an Ab-VAPAB- PEG-mExa structure having a cleavable VAPAB linker between the antibody and the polymer, such as a PEG (FIG. 7H). Although the term “mExa” is specifically used, any PBA disclosed herein, or known to those skilled in the art can be used.

[0173] In some cases, the pharmaceutical composition can comprise a formula selected from Formula ID 50, 51 , 52, 53, 54, 55, 56, 57, 58, 59, 60, 61 , 62, 63, 64, 65, and a combination thereof, as disclosed hereafter.

[0174] Representative examples of cleavable linkers can be selected from Formula ID 101-141 , or a combination thereof.

[0175] In some cases, the pharmaceutical composition can comprise at least a branched or linear polymer side chain. In some cases, schematic illustrations of representative examples of the pharmaceutical composition can comprise at least a branched or linear polymer side chain that has one or more free second terminal groups that each is free from PBAs are shown in FIG. 8A-FIG. 8L. In some cases, a pharmaceutical composition of this disclosure can comprise a Linker L1 (2a), a Linker L2 (2b) or a combination thereof, without any cleavable linkers between the TBA (1), polymer (4) and first PBA (3), and a branched polymer side chain (4’) attached to the polymer (4) (FIG. 8A). In some cases, a pharmaceutical composition can comprise a Linker L1 (2a), a Linker L2 (2b) or a combination thereof, without any cleavable linkers between the TBA (1), polymer (4) and first PBA (3), and a branched polymer side chain (4’) attached to the TBA (1) (FIG. 8B). In some cases, a pharmaceutical composition can comprise at least one cleavable linker, such as the first cleavable linker (8a), the second cleavable linker (8b), or a combination thereof, and a branched polymer side chain (4’) attached to the polymer (FIG. 8C). In some cases, a pharmaceutical composition can comprise at least one cleavable linker, such as the first cleavable linker (8a), the second cleavable linker (8b), or a combination thereof, and a branched polymer side chain (4’) attached to the TBA (FIG. 8D). In some cases, a pharmaceutical composition can comprise a Linker L1 (2a), a Linker L2 (2b) or a combination thereof, without any cleavable linkers between the TBA (1), polymer (4) and first PBA (3), and a linear polymer side chain (4”) attached to the polymer (4) (FIG. 8E). In some cases, a pharmaceutical composition can comprise a Linker L1 (2a), a Linker L2 (2b) or a combination thereof, without any cleavable linkers between the TBA (1),polymer (4) and first PBA (3), and a branched polymer side chain attached to the TBA (1 ) (FIG. 8F). In some cases, a pharmaceutical composition can comprise at least one cleavable linker, such as the first cleavable linker (8a), the second cleavable linker (8b), or a combination thereof, and a linear polymer side chain (4”) attached to the polymer (4) (FIG. 8G). In some cases, a pharmaceutical composition can comprise at least one cleavable linker, such as the first cleavable linker (8a), the second cleavable linker (8b), or a combination thereof, and a linear polymer side chain (4”) attached to the TBA (1) (FIG. 8H). In some cases, a pharmaceutical composition can comprise at least one cleavable linker and a linear or branched polymer side chain (4’) attached to the linker, such as the Linker L1 (8a) that is cleavable (FIG. 8I). In some cases, a pharmaceutical composition can comprise at least one cleavable linker and a linear or branched polymer side chain (4’) attached to the linker, such as the Linker L2 that is cleavable (8b) (FIG. 8J). In some cases, a pharmaceutical composition can comprise at least one cleavable linker and a linear polymer side chain (4’) attached to the linker, such as the Linker L1 (2a) that is not cleavable (FIG. 8K). In some cases, a pharmaceutical composition can comprise at least one cleavable linker and a linear polymer side chain (4’) attached to the linker, such as the Linker L2 (2b) that is not cleavable (FIG. 8L). R1 and R2 represent the first terminal group and the second terminal group, respectively, as disclosed herein.

[0176] In some cases, the pharmaceutical composition can comprise at least a branched or linear polymer side chain that is attached to at least a PBA. In some cases, schematic illustrations of representative examples of the pharmaceutical composition can comprise at least a branched or linear polymer side chain that has one or more one second terminal group that is attached to at least a PBA are shown in FIG. 9A-FIG. 9L. In some cases, a pharmaceutical composition of this disclosure can comprise a Linker L1 (2a), a Linker L2 (2b) ora combination thereof, without any cleavable linkers between the TBA (1), polymer (4) and first PBA (3), and a branched polymer side chain (4’) attached to the polymer (4) and at least a second PBA (3’) (FIG. 9A). In some cases, a pharmaceutical composition can comprise a Linker L1 (2a), a Linker L2 (2b) or a combination thereof, without any cleavable linkers between the TBA (1), polymer (4) and first PBA (3), and a branched polymer side chain (4’) attached to the TBA (1) and at least a second PBA (3’) (FIG. 9B). In some cases, a pharmaceutical composition can comprise at least one cleavable linker, such as the first cleavable linker (8a), the second cleavable linker (8b), or a combination thereof, and a branched polymer side chain (4’) attached to the polymer and at least a second PBA (3’) (FIG. 9C). In some cases, a pharmaceutical composition can comprise at least one cleavable linker, such as the first cleavable linker (8a), the second cleavableIl llinker (8b), or a combination thereof, and a branched polymer side chain (4’) attached to the TBA and at least a second PBA (3’) (FIG. 9D). In some cases, a pharmaceutical composition can comprise a Linker L1 (2a), a Linker L2 (2b) or a combination thereof, without any cleavable linkers between the TBA (1), polymer (4) and first PBA (3), and a linear polymer side chain (4”) attached to the polymer (4) and at least a second PBA (3’) (FIG. 9E). In some cases, a pharmaceutical composition can comprise a Linker L1 (2a), a Linker L2 (2b) or a combination thereof, without any cleavable linkers between the TBA (1), polymer (4) and first PBA (3), and a branched polymer side chain attached to the TBA (1) and at least a second PBA (3’) (FIG. 9F). In some cases, a pharmaceutical composition can comprise at least one cleavable linker, such as the first cleavable linker (8a), the second cleavable linker (8b), or a combination thereof, and a linear polymer side chain (4”) attached to the polymer (4) and at least a second PBA (3’) (FIG. 9G). In some cases, a pharmaceutical composition can comprise at least one cleavable linker, such as the first cleavable linker (8a), the second cleavable linker (8b), or a combination thereof, and a linear polymer side chain (4”) attached to the TBA (1) and at least a second PBA (3’) (FIG. 9H). In some cases, a pharmaceutical composition can comprise at least one cleavable linker and a linear or branched polymer side chain (4’) attached to at least one PBA and the linker, such as the Linker L1 (8a) that is cleavable (FIG. 9I). In some cases, a pharmaceutical composition can comprise at least one cleavable linker and a linear polymer side chain (4’) attached to at least a PBA and the linker, such as the Linker L2 (8b) that is cleavable (FIG. 9J). In some cases, a pharmaceutical composition can comprise at least one cleavable linker and a linear or branched polymer side chain (4’) attached to at least a PBA and the linker, such as the Linker L1 (2a) that is not cleavable (FIG. 9K). In some cases, a pharmaceutical comprising at least one cleavable linker and a linear or branched polymer side chain (4’) attached to at least a PBA and the linker, such as the Linker L2 (2b) that is not cleavable (FIG. 9L). R1 and R2 represent the first terminal group and the second terminal group, respectively, as disclosed herein. In some cases, the pharmaceutical composition can comprise at least one of the R1 , R2 or a combination thereof of the polymer side chain, free from the PBAs. In some cases, the pharmaceutical composition can comprise at least one of the R1 , R2 or a combination thereof, of the polymer side chain, attached to one or more PBAs, such as the first PBA, the second PBA, or a combination thereof. The first PBA (3) and the second PBA (3’) can be the same or different, and each can be independently selected form the PBAs disclosed herein, or a combination thereof. For simplicity, a linear or a branched polymercan be shown in some figure and can represent a linear polymer, a branched polymer or a combination thereof.

[0177] In some cases, each of the two or more branches can be the polymer side chain and can be attached to one or more PBAs.

[0178] In some cases, the pharmaceutical composition can comprise an antibody, polymer, PBA, one or more linkers, and optionally, a coupler, or a combination thereof. In some cases, an antibody can be covalently linked to a PBA directly (Formula ID 50). In some cases, an antibody can be covalently linked to a polymer and a PBA wherein the polymer is positioned between the antibody and the PBA (Formula ID 51 ). In some cases, an antibody can be covalently linked to a linker and a PBA wherein the linker is positioned between the antibody and the PBA (Formula ID 52). In some cases, an antibody can be covalently linked to a polymer, a linker and a PBA wherein the polymer is linked to the antibody and the linker, and the linker is positioned between the polymer and the PBA (Formula ID 53). In some cases, an antibody can be covalently linked to a linker and the linker is linked to the polymer, wherein the polymer is positioned between the linker and the PBA (Formula ID 54). In some cases, an antibody can be covalently linked to one or more linkers that can include a Linker L1 , a Linker L2 or a combination thereof, wherein the Linker L1 is positioned between the antibody and the polymer, and the Linker L2 is positioned between the polymer and the PBA (Formula ID 55). In some cases, an antibody can be covalently linked to a coupler and a PBA wherein the coupler is positioned between the antibody and the PBA (Formula ID 56). In some cases, an antibody can be covalently linked to a coupler, a linker and a PBA wherein the coupler is linked to the antibody and the linker, and the linker is positioned between the coupler and the PBA (Formula ID 57). In some cases, an antibody can be covalently linked to a coupler, the coupler is linked to the polymer, and the polymer is linked to the linker that is linked to the PBA (Formula ID58). In some cases, an antibody can be covalently linked to a coupler, the coupler is linked to the linker, and the linker is linked to the polymer that is linked to the PBA (Formula ID59).

[0179] In some cases, an antibody can be covalently linked to one or more linkers that can include a Linker L1 , a Linker L2 or a combination thereof, wherein the coupler is linked to the antibody and the Linker L1 that is linked to the polymer linked to the Linker L2 and then linked to the PBA (Formula ID 60). In some cases, an antibody can be covalently linked to one or more linkers that can include a Linker L1 , a Linker L2 or a combination thereof, and linked to the two or more PBAs. In some cases, an Nano-ADC can comprise an antibody covalently linked to two or more same or different PBAs, such as Formula ID61-65, Formula ID 3033, 3034, 3035, 3036, 3044, 3066, 3067, 3068, 3069, or a combination thereof.

[0180] In some cases, an antibody can be covalently linked to one or more PBAs via sitespecific enzymatic reactions, such as those commercially available ADC conjugation kits.

[0181] In some cases, disulfide bonds of an antibody can be partially reduction with either dithiothreitol (DTT) or tris(2-carboxyethyl) phosphine (TCEP) to release free Cys for drug conjugation. In some cases, 5,5'-dithiobis (2-nitrobenzoic acid) (DTNB) can be used to produce drug conjugates connect to the Cys residues usually involved in heavy-heavy inter-chain disulfides.

[0182] In some cases, cysteine-based conjugation can involve the mutation of some hinge region cysteine residues to serine that seems to have no significant influence on antibody binding affinity, in vitro conjugate stability and not to induce antibody aggregation.

[0183] In some cases, the pharmaceutical composition can comprise a formula selected from Formula ID 50, 51 , 52, 53, 54, 55, 56, 57, 58, 59, 60, 61 , 62, 63, 64, 65, and a combination thereof:wherein the antibody (Ab) can be selected from alemtuzumab, anti-CD19 antibodies, anti-CD20 antibodies, anti-CTLA-4 (cytotoxic T lymphocyte-associated antigen) antibodies, anti-LAG3 (lymphocyte activation gene-3) antibodies, anti-PD1 antibodies KEYTRUDA® (pembrolizumab), anti-PD-L1 antibodies, anti-TIM-3 (T cell immunoglobulin and mucin domain-3) antibodies, atezolizumab, BAVENCIO® (avelumab), bevacizumab, blinatumomab, brentuximab, cetuximab, daratumumab, datopotamab, denosumab, dinutuximab, disitamab, durvalumab, elotuzumab, enfortumab, gemtuzumab, ibritumomab, IMFINZI® (durvalumab), inotuzumab, ipilimumab, lectins, loncastuximab, mirvetuximab, necitumumab, obinutuzumab, ofatumumab, olaratumab, OPDIVO® (nivolumab), panitumumab, pembrolizumab, pertuzumab (anti-HER2), polatuzumab, ramucirumab, rovalpituzumab, rituximab, rituximab, sacituzumab, siltuximab, toripalimab, tislelizumab (BGB-A317), tisotumab, TECENTRIQ® (atezolizumab), tositumomab, trastuzumab (anti-HER2), one or more antibodies against target molecules comprising 5T4, ALPPL2, AXL, B7H3, B7H4, B7H6, B7S1 , B7x, BCAM, BCMA, CCL2, CCL5, CD146, CD25, CD27 ligand, CD276, CD47, CD70, CDH3, CDH6, CEACAM5, CEACAM6, CLDN1 , CLDN18.2, CLDN6, DLK1 , DLL3, EGFR, EGFRVIII, EpCAM, ErbB1 , ErbB2, ErbB3, FAT2, FOLR1 , GPC3, GPR87, GRM8, GRP56, HER2, HER3, HGFR, ITGB6, LIV-1 , Ly6G6D, LY6G6F, LYPD3, MCAM, MET, MSLN, MUC16, MUC18, PSMA, ROR1 , SEZ6, SLC39A6, SLC7A11 , SLCO1 B3, STEAP1 , Tacstd2, TLR4, TMPRSS4, TROP2, UFO, VEGF, VTCN1 , ZIP6, a bispecific antibody thereof, a tri-specific antibody thereof, a tetra-specific antibody thereof, a fragment thereof, an antigen-binding portion thereof, and a combination thereof; the coupler can be selected from Formula ID 201 , 202, 203, 204, 205, 206, 207, 208, 209, and a combination thereof; the polymer can be selected from a peptide, a polyethylene glycol (PEG), polyoxazoline (POX), or a combination thereof, wherein the polyoxazoline (POX) comprises a linear portion, a branched portion, or a combination thereof, and wherein the polyoxazoline (POX) comprises poly(2-methyloxazoline), poly(2-ethyloxazoline), poly(2- propyloxazoline), poly(2-isopropyloxazoline), poly(2-substituted oxazoline), and a combination thereof; and the PBA can be selected from Formula ID 1 , 1-1 , 1-2, 2, 2-1 , 2-2, 3, 3-1 , 3-2, 1001 , 1002, 1003, 1004, 1005, 1006, 1007, 1008, 1009, 1010, 1011 , 1012, 1013, 1014, 1015,1016, 1017, 1018, 1019, 1020, 1021 , 1022, 1023, 1024, 1025, 1026, 1027, 1028, 1029,1030, 1031 , 1032, 1033, 1034, 1035, 1036, 1037, 1038, 1039, 1040, 1041 , 1042, 1043,1044, 1045, 1046, 1047, 1048, 1049, 1050, 1051 , 1052, 1053, 1054, 1055, 1056, 1057,1058, 1059, 1060, 1061 , 1062, 1063, 1064, 1065, 1066,1067, 1068, 1069, 1070, 1071 ,1072, 1073, 1074, 1075, 1076, 1077, 1078, 1079, 1080, 1081 , 1082, 1083, 1084, 1085,1086, 1087, 1088, 1089, 1090, 1091 , 1092, 1093, 1094, 1095, 1096, 1097, 1098, 1099,1100, 1 101 , 1102, 1103, 1 104, 1105, 1106, 1107, 1108, 1109, 1110, 1111 , 1112, 11 13,1114, 1 115, 1116, 1117, 1 118, 1119, 1120, 1121 , 1122, 1123, 1124, 1 125, 1126, 1127,1 128, 1129, 1 130, 1131 , 1132, 1133, 1134, 1135, a pharmaceutically acceptable salt thereof, a solvate thereof, a prodrug thereof, an isomer thereof, an isotope derivative thereof, and a combination thereof; and n is an integer and can be in a range of from 1 to 100.

[0184] PBA1 and PBA2 can be a first and a second PBA, respectively, and can be the same or different. In some cases, the pharmaceutical composition can comprise Formula ID 2005, 2027, 2028, 2029, 2030, 2037 or a combination thereof.

[0185] In some cases, n can be in a range of from 1 to 24, 1 to 20, 1 to 18, 1 to 16, 1 to 14, 1 to 12, 1 to 10, 1 to 8, 1 to 6, 1 to 4, or 1 to 2. An antibody, such as an IgG, can have multiple cystine residues that can provide cystine-based conjugations resulting in multiple couplers, linkers, polymers or PBAs linked to a single antibody. In some cases, n can be 2, 4, 6, or 8.

[0186] In some cases, the linker, Linker L1 and Linker L2, when present, each can be independently a cleavable linker and can be independently selected from Formula ID 101 , 102, 103, 104, 105, 106, 107, 108, 109, 110, 111 , 112, 1 13, 114, 115, 116, 117, 118, 119, 120, 121 , 122, 123, 124, 125, 126, 127, 128, 129, 130, 131 , 132, 133, 134, 135, 136, 137, 138, 139, 140, 141 , and a combination thereof.

[0187] In some cases, the pharmaceutical composition disclosed herein can comprise a TBA that comprises an antibody having binding affinity to a target molecule that comprises a tumor antigen (Ag). In some cases, the pharmaceutical composition can comprise tumor cytotoxicity comprising AgHightumor cytotoxicity to tumor cells having a tumor antigen or a high level of the tumor antigen, sometimes referred to as abnormally high level of tumor antigen (collectively, AgHightumor cells) and AgLowtumor cytotoxicity to tumor cells having low, very low or free from the tumor antigen (collectively, AgLowtumor cells). In some cases, the pharmaceutical composition can comprise AgLowtumor cytotoxicity to AgLowtumor cells in the absence of the AgHightumor cells.

[0188] In some cases, the TBA can be selected from Alemtuzumab, Atezolizumab, Avelumab, bevacizumab, Blinatumomab, Brentuximab, cetuximab, Daratumumab, Denosumab, Dinutuximab, Durvalumab, Elotuzumab, GEMTUZUMAB, Ibritumomab, Inotuzumab, Ipilimumab, Necitumumab, Nivolumab, Obinutuzumab, Ofatumumab, Olaratumab, Panitumumab, Pembrolizumab, Pertuzumab, Ramucirumab, Rituximab, Siltuximab, Tislelizumab (BGB-A317), Toripalimab, Trastuzumab, and a combination thereof.

[0189] In particular cases, the TBA can be selected from Pertuzumab, Trastuzumab, a biosimilar antibody, and a combination thereof. In some cases, the pharmaceutical composition can comprise AgHightumor cytotoxicity to HER2+tumor cells and AgLowtumor cytotoxicity to HER2' tumor cells, wherein AgLowtumor cytotoxicity can be measured in a cell-based assay with HER2' tumor cells in the absence of any HER2+tumor cells or measured using tumor size in tumor cell xenografted mouse models (as described in Examples and FIG. 17 and FIG. 18).

[0190] In some cases, polymer can comprise a single amino acid residue, dipeptide, tripeptide, or polypeptide. In some cases, as mentioned above, the polymer can comprise a peptide comprising in a range of from 1 to 100 amino acid residues. In some cases, a polymer can comprise a lysine, glutamic acid, glutamine, arginine, valine, or a combination thereof.

[0191] In some cases, the pharmaceutical composition can comprise at least a polymer side chain that is covalently linked to the TBA, the polymer, or a combination thereof, wherein the polymer side chain comprises POX comprising poly(2-methyloxazoline), poly(2-ethyloxazoline), poly(2-propyloxazoline), poly(isopropyloxazoline), lysine, arginine, glutamic acid, glutamine, a reacted form thereof, a derivative thereof, or a combination thereof.

[0192] In some cases, the pharmaceutical composition can comprise a TBA, a PBA and a polymer comprising one or more branches having reactive terminal groups selected from -NH2, -OH, -SH, -COOH, -CO, or a combination thereof. In some cases, the polymer can comprise a lysine (in a reacted form, i.e. , a lysine residue) and a polymer side chain can be reacted and covalently linked to an -NH2 of the polymer. In some cases, the polymer can comprise one or more amino acid residues and a polymer side chain can be covalently linked to the amino acid residue.

[0193] In some cases, the pharmaceutical composition can comprise one or more polymer side chains and at least one of the polymer side chains can comprise a covalently linked PBA. In some cases, each of the polymer side chains can comprise a covalently linkedPBA. The PBA can comprise the aforementioned Formula ID 1 , 1-1 , 1-2, 2, 2-1 , 2-2, 3, 3- 1 , 3-2, and 1001-1135, the additional cytotoxic agent, a pharmaceutically acceptable salt thereof, a solvate thereof, a prodrug thereof, an isomer thereof, an isotope derivative thereof, a reacted PBA thereof, or a combination thereof.

[0194] In some cases, the pharmaceutical composition can comprise one or more mExa selected from Formula ID 1 , 1-1 , 1-2, 2, 2-1 , 2-2, 3, 3-1 , 3-2, and 1001-1135, and a combination thereof. In some cases, the pharmaceutical composition can comprise one or more mExa selected from Formula ID 1 , 1-1 , 1-2, 2, 2-1 , 2-2, 3, 3-1 , 3-2, and 1001- 1135, exatecan, deruxtecan, deruxtecan derivatives, MMAE, MMAF, and a combination thereof.

[0195] Some unlimited examples can include aforementioned Formula ID 2021 , 2022, 2023, 2024, 2025, 2026, 2027, 2028, 2029, 2030, 2031 , 2032, 2033, 2034, 2035, 2036, 2037, 2038, 2039 or 2040 (The Formulas are illustrated as molecules prior to reacting, i.e., linking to a TBA).

[0196] In some cases, the polymer can comprise polyoxazoline (POX) having a branched portion that can comprise two or more branches, and wherein at least one of the two or more branches can be the polymer side chain and can be free from the PBA. In some cases, each of the two or more branches can be the polymer side chain and can be attached to one or more PBAs.

[0197] In some cases, the pharmaceutical composition can comprise a formula selected from Formula ID 50-62, or a combination thereof, and a polymer side chain that is covalently linked to the Ab, the polymer, or a combination thereof, wherein the polymer side chain can comprise POX comprising poly(2-methyloxazoline), poly(2- ethyloxazoline), poly(2-propyloxazoline), poly(isopropyloxazoline), or a combination thereof.

[0198] In some cases, the PBA can comprise the Formula ID 1 , 1-1 , 1-2, 2, 2-1 , 2-2, 3, 3- 1 , 3-2, and 1001-1135, calicheamicin, DM1 (antimicrotubule agent T-DM1 , a derivative of maytansine), DM4 (Ravtansine), Dxd (deruxtecan), exatecan, IRDye700DX (a nearinfrared (NIR) photosensitizer suitable for near infrared (NIR) photoimmunotherapy (PIT)), MMAE (Monomethyl auristatin E, also known as vedotin), MMAF (Monomethyl auristatin F), pseudomonas exotoxin A (a potent exotoxin produced by the bacterium Pseudomonas aeruginosa), pyrrolobenzodiazepine (PBD), SG3199 (a cytotoxic DNA minor groove interstrand crosslinking pyrrolobenzodiazepine (PBD) dimer), SN-38, irinotecan, camptothecin, amanitin (an inhibitor of RNA Polymerase II), duocarmycins (DNA- alkylators), halichondrin (Eribulin, also known Halaven®, under respective trademark),norhalichondrin B, maytansinoids, maitansine (INN), maytansine, PUN-159682 (an inhibitor of DNA Topoisomerase II), tubulysins, tubulysin, pretubulysin (PT), tubulysin A (TubA), tubulysin M, or a combination thereof.

[0199] In some cases, the pharmaceutical composition can comprise the polymer that comprises polyoxazoline (POX), wherein the polyoxazoline (POX) can comprise a linear portion, a branched portion, or a combination thereof, and wherein the polyoxazoline (POX) comprises poly(2-methyloxazoline), poly(2-ethyloxazoline), poly(2- propyloxazoline), poly(2-isopropyloxazoline), poly(2-substituted oxazoline), and a combination thereof, polyoxazoline (POX) having a branched portion that can comprise two or more branches, and one or more PBAs selected from the PBA having the Formula ID 1001-1135, calicheamicin, DM1 (antimicrotubule agent T-DM1 , a derivative of maytansine), DM4 (Ravtansine), Dxd (deruxtecan), exatecan, IRDye700DX (a nearinfrared (NIR) photosensitizer suitable for near infrared (NIR) photoimmunotherapy (PIT)), MMAE (Monomethyl auristatin E, also known as vedotin), MMAF (Monomethyl auristatin F), pseudomonas exotoxin A (a potent exotoxin produced by the bacterium Pseudomonas aeruginosa), pyrrolobenzodiazepine (PBD), SG3199 (a cytotoxic DNA minor groove interstrand crosslinking pyrrolobenzodiazepine (PBD) dimer), SN-38, irinotecan, camptothecin, amanitin (an inhibitor of RNA Polymerase II), duocarmycins (DNA- alkylators), halichondrin (Eribulin, also known Halaven®, under respective trademark), norhalichondrin B, maytansinoids, maitansine (INN), maytansine, PUN-159682 (an inhibitor of DNA Topoisomerase II), tubulysins, tubulysin, pretubulysin (PT), tubulysin A (TubA), tubulysin M, a pharmaceutically acceptable salt thereof, a solvate thereof, a prodrug thereof, an isomer thereof, an isotope derivative thereof, and a combination thereof.

[0200] In some cases, the PBA can be selected from Formula ID 1 , 1-1 , 1-2, 2, 2-1 , 2-2, 3, 3-1 , 3-2, 1001 , 1002, 1003, 1004, 1005, 1006, 1007, 1008, 1009, 1010, 1011 , 1012, 1013, 1014, 1015, 1016, 1017, 1018, 1019, 1020, 1021 , 1022, 1023, 1024, 1025, 1026,1027, 1028, 1029, 1030, 1031 , 1032, 1033, 1034, 1035, 1036, 1037, 1038, 1039, 1040,1041 , 1042, 1043, 1044, 1045, 1046, 1047, 1048, 1049, 1050, 1051 , 1052, 1053, 1054,1055, 1056, 1057, 1058, 1059, 1060, 1061 , 1062, 1063, 1064, 1065, 1066,1067, 1068, 1069, 1070, 1071 , 1072, 1073, 1074, 1075, 1076, 1077, 1078, 1079, 1080, 1081 , 1082,1083, 1084, 1085, 1086, 1087, 1088, 1089, 1090, 1091 , 1092, 1093, 1094, 1095, 1096,1097, 1098, 1099, 1100, 1 101 , 1102, 1103, 1104, 1105, 1106, 1107, 1 108, 1109, 1110,1 111 , 1 112, 1113, 1114, 1 115, 1116, 1117, 1118, 1119, 1120, 1121 , 1122, 1123, 1124,1 125, 1 126, 1127, 1128, 1129, 1 130, 1131 , 1132, 1 133, 1134, 1135, a pharmaceuticallyacceptable salt thereof, a solvate thereof, a prodrug thereof, an isomer thereof, an isotope derivative thereof, and a combination thereof.

[0201] The pharmaceutical composition disclosed herein, wherein the Ab can be covalently linked to the PBA, the coupler, the polymer, the linker, the Linker L1 , the Linker L2, or a combination thereof, to form an antibody-drug conjugate (ADC) having a formula selected from Formula ID 50-62, or a combination thereof.

[0202] In some cases, the Ab can be covalently linked to a biomolecule having a formula selected from Formula ID 2001 , 2002, 2003, 2004, 2005, 2006, 2007, 2008, 2009, 2010, 2011 , 2012, 2013, 2014, 2015, 1026, 2017, 2018, 2019, 2020, 2021 , 2022, 2023, 2024, 2025, 2026, 2027, 2028, 2029, 2030, 2031 , 2032, 2033, 2034, 2035, 2036, 2037, 2038, 2039, 2040, a pharmaceutically acceptable salt thereof, a solvate thereof, a prodrug thereof, an isomer thereof, an isotope derivative thereof, and a combination thereof, to form an antibody-drug conjugate (ADC).

[0203] In some cases, the linker, Linker L1 and Linker L2, when present, each can be independently a cleavable linker and can be independently selected from Formula ID 101 , 102, 103, 104, 105, 106, 107, 108, 109, 110, 111 , 112, 1 13, 114, 115, 116, 117, 118, 119, 120, 121 , 122, 123, 124, 125, 126, 127, 128, 129, 130, 131 , 132, 133, 134, 135, 136, 137, 138, 139, 140, 141 , and a combination thereof.

[0204] In some cases, a cystine residue of an antibody can be reacted to a cystine-based coupler (also referred to as “cystine-based conjugation”) such as succinimidyl-4-(N- maleimidomethyl)cyclohexane-1-carboxylate (SMCC) (Formula ID 201), maleimide (Mai) (Formula ID 202), maleimidocaproyl (MC) (Formula ID 203), maleimidomethyl cyclohexane-1 -carboxylate (MCC) (Formula ID 204), or dibromomaleimide (DBM) (Formula ID 205).

[0205] In some cases, an antibody can be linked to one or more, same or different biomolecules having a formula selected from Formula ID 2001-2040, a pharmaceutically acceptable salt thereof, a solvate thereof, a prodrug thereof, an isomer thereof, an isotope derivative thereof, and a combination thereof, via one or more cystine residues to form an antibody-drug conjugate (ADC).

[0206] It is understood that the coupler, the linker, the polymer and the PBA each can be in a reacted form covalently linked together in the pharmaceutical composition disclosed herein, such as the aforementioned cystine-based conjugation.

[0207] The pharmaceutical composition can comprise the AgLowtumor cytotoxicity to AgLowtumor cells in the absence of any AgHightumor cells. In another word, the AgLowtumor cytotoxicity of the Nano-ADC of this disclosure to AgLowtumor cells can be independent of any AgHightumor cells.

[0208] For a traditional ADC, the drug, i.e. , a PBA, from an ADC can be released either from the target cell following internalization and degradation of the ADC or in the extracellular space. The drug can then be taken up by surrounding or bystander cells killing these cells resulting in “bystander cytotoxicity”. These surrounding or bystander cells themselves may or may not express the ADC target antigen, i.e., can be AgLowtumor cells or AgLownormal cells. Traditional ADCs, such as the aforementioned ADC famtrastuzumab deruxtecan (ENHERTU® ADC), exhibit no AgLowtumor cytotoxicity to AgLowtumor cells in the absence of any AgHightumor cells.

[0209] The AgLowtumor cytotoxicity, the AgHightumor cytotoxicity, and the non-tumor cytotoxicity, each can be measured in a cell-based assay. In one example, a cell viability assay can be used. In another example, a cell death assay can be used. Commercial assay kits, such as those available from Thermo Fisher Scientific can be suitable.

[0210] In any of suitable cell-based assays, only AgLowtumor cells can be used in the absence of any AgHightumor cells. Traditional ADCs, such as ENHERTU ADC having a monoclonal antibody trastuzumab and a cytotoxic drug (PBA) deruxtecan, exhibit no such AgLowtumor cytotoxicity in the absence of any AgHightumor cells.

[0211] Multiple methods can be employed to calculate the differences in cytotoxicity. One way to calculate a difference (in percentage number) of the AgLowtumor cytotoxicity (Viability %: VA9NT) V.S. the non-tumor cytotoxicity (Viability %: Vnormai) can be based on cell viability using the following formula: difference=VA9NT - Vnormai.

[0212] In some cases, the Nano-ADC of this disclosure can exhibit AgLowtumor cytotoxicity at least 20%, 30%, 40%, 50% higher than that of the non-tumor cytotoxicity, i.e., VA9NT is at least 20%, 30%, 40%, 50% lower than Vnormai if a cell viability is used. In other words, the Nano-ADC of this disclosure can exhibit AgLowtumor cytotoxicity with a cell death rate at least 20%, 30%, 40%, 50% higherthan that of the non-tumor cytotoxicity, when a cell death rate is used.

[0213] In some cases, the Nano-ADC of this disclosure can exhibit AgLowtumor cytotoxicity at least 20%, 30%, 40%, 50% higher than that of the non-tumor cytotoxicity, i.e., VA9NT is at least 20%, 30%, 40%, 50% lower than Vnormai when tumor sizes are measured. In other words, the Nano-ADC of this disclosure can exhibit AgLowtumor cytotoxicity with tumor size reduction at least 20%, 30%, 40%, 50% higher than that of the control, when tumor size measurement is used.

[0214] In some cases, since each of the traditional ADCs is configured to have a mAb targeting a tumor antigen (Ag) on tumor cells, a cell-based assay using AgLowtumor cells can be used to measure the AgLowtumor cytotoxicity. Not wishing to be bound by any particular calculation methods, in some cases, Applicants use cell viability to calculate cytotoxicity. For example, specific AgLowtumor cells, such as MDA-MB231 cells of a triplenegative breast cancer (TNBC) (HER2-) cell line can be used to assay the AgLowtumor cytotoxicity of the ADC comprising an anti-HER2 antibody. In such assays, a traditional ADC, such as ENHERTU® ADC can be used as a control to compare with the cytotoxicity of the Nano-ADC of this disclosure and the cytotoxicity of ENHERTU® towards HER2' cells, such as the MDA-MB231 cells or other non-HER2 overexpressing cells.

[0215] In some cases, the polymer can be covalently linked to the TBA and the PBA, wherein the polymer comprises a reacted first terminal group and a reacted second terminal group; and wherein the polymer is positioned between the TBA and the PBA with the reacted first terminal group covalently linked to the TBA and the reacted second terminal group covalently linked to the PBA.

[0216] In some cases, the pharmaceutical composition can further comprise a Linker L1 , a Linker L2, or a combination thereof, wherein the Linker L1 can be covalently linked to the TBA and covalently linked to the reacted first terminal group of the polymer, the Linker L2 can be covalently linked to the PBA and covalently linked to the reacted second terminal group of the polymer, with a proviso that the Linker L1 and the Linker L2 can each be covalently linked to TBA or PBA when the polymer is absent.

[0217] In some cases, at least one of the Linker L1 and the Linker L2 comprises a cleavable linker that is cleavable in vivo or in the target cell. In some cases, the cleavable linker can comprise an enzymatically-cleavable peptide linker, GGFG linker, VAPAB linker, VCPAB linker, glutamic acid-glycine-citrulline (EGCit) linkers, acid sensitive hydrazone linker, a linker comprising disulfides, a thioether linker, a glutathione-sensitive disulfide linker, a linker containing a sugar molecule or moiety, a glucuronide containing linker, a glycosidase cleavable linker, a phosphatase cleavable linker, an esterase cleavable linker, a hydrolysis cleavable linker, a modified monodispersed PEG linker, a modified PEG linker, a modified poly-PEG linker, a dipeptide linker, a valine— citrulline linker, a valine-alanine linker, or a combination thereof. In some cases, a PEOX polymer comprising disulfide bonds can also be suitable as a cleavable linker.

[0218] In some cases, the enzymatically-cleavable peptide linker comprises a GGFG linker, a VAPAB linker, or a combination thereof. Commercial linkers, such as Mc-Gly- Gly-Phe-Gly-PAB-OH (Mc-GGFG-PAB-OH), Gly-Gly-L-Phe-N-[(carboxymethoxy)methyl]Glycinamid (GGFG-NCM) linker, L-Val-L-Ala-para- aminobenzyl alcohol (VAPAB-OH) linker, or L-Val-L-Cit-para-aminobenzyl alcohol (VCPAB-OH) linker can be suitable.

[0219] In some cases, each of the Linker L1 , the Linker L2, or a combination thereof, can comprise the cleavable linker. In some cases, each of the Linker L1 , the Linker L2, or a combination thereof, comprises the enzymatically-cleavable peptide linker comprising a GGFG linker, a VAPAB linker, a VCPAB linker, or a combination thereof.

[0220] In some cases, the cleavable linker can comprise one or more glutamic acid- glycine-citrulline (EGCit) linkers.

[0221] In some cases, the cleavable linker can comprise at least one GGFG linker, VAPAB Linker, VCPAB linker, or a combination thereof. In some cases, the cleavable linker can comprise one or more GGFG linkers, VAPAB Linkers, VCPAB linker, or a combination thereof.

[0222] In some cases, the linker, the Linker L1 , the Linker L2, or a combination thereof, each can independently comprise a cleavable linker selected from the Formula ID 1 QI- 141 , or a combination thereof.

[0223] In some cases, a small molecule binding pair can be any one of binding pairs that can comprise a member being a small molecule and a member of either a small or a large molecule. Members of such a binding pair can bind to each other under proper conditions. Examples of small molecule binding pairs can include inhibitor-target binding pair, stimulator-effector binding pair, biotin -streptavidin binding pair, avidin- streptavidin / neutravidin binding pair, dinitrophenol (DNP) and an anti-DNP antibody binding pair, a digoxin and an anti-digoxin antibody binding pair, a digoxigenin and an anti-digoxigenin antibody binding pair, a hapten and an anti-hapten antibody binding pair, a polysaccharide and a polysaccharide binding moiety, such as a lectin, or a combination thereof.

[0224] In some cases, the PBA can comprise a natural or synthetic small molecule-based drug, inorganic-based drug, a toxin molecule having cytotoxicity, a molecule having cytotoxicity to the target cell, a derivative thereof, or a combination thereof.

[0225] In some cases, the PBA can comprise any of the PBAs disclosed herein.

[0226] In some cases, the pharmaceutical composition can comprise a molar ratio of the TBA to the PBA in a range of from about 1 :0.01 to about 1 : 100, 1 :0.1 to about 1 : 100, 1 : 1 to about 1 :100. In particular cases, the pharmaceutical composition can comprise a molar ratio of the TBA to the PBA in a range of from 1 :1 to about 1 :100, 1 :1 to about 1 :80, 1 :1 to about 1 :70, 1 :1 to about 1 :60, 1 : 1 to about 1 :50, 1 :1 to about 1 :40, 1 : 1 to about 1 :30,1:1 to about 1:20, 1:1 to about 1:10, 1:1 to about 1:5, 1:1 to about 1:4, 1:1 to 1:3, 1:1 to 1:2.

[0227] In some cases, the pharmaceutical composition can comprise a molar ratio of the TBA to the PBA in a range of 1:2 to 1:100. In particular cases, the pharmaceutical composition can comprise a molar ratio of the TBA to the PBA in a range of from 1:2 to about 1:100, 1:2 to about 1:80, 1:2 to about 1:70, 1:2 to about 1:60, 1:2 to about 1:50, 1:2 to about 1 :40, 1:2 to about 1:30, 1:2 to about 1 :20, 1 :2 to about 1:10, 1 :2 to about 1:5, 1 :2 to about 1 :4, and 1 :2 to 1 :3.

[0228] In some cases, the pharmaceutical composition can comprise a mixture of TBA, wherein some of the TBA can be covalently linked to the PBA and some of the TBA can be non-covalently mixed with the PBA leading to more TBAs than the PBAs. In some cases, some antibody molecules can be free from bound drugs, i.e., free antibody without conjugated to a drug molecule. In some cases, the molar ratio of TBA:PBA can be 100:1, 20:1, 10:1, 5:1, 4;1, 3;1, or 2:1 (on a reverse ratio of 1:0.01, 1:0.1, 1:0.2, 1:0.25, 1:0.3, and 1:0.5). In some cases, the molar ratio of antibody to a drug can be 100: 1 , 10:1, 5:1, 4;1, 3; 1 , or2:1 (on a reverse ratio of 1:0.01, 1:0.1, 1:0.2, 1:0.25, 1:0.3, and 1:0.5).

[0229] In some cases, the pharmaceutical composition can comprise one molecule of TBA and at least 2 molecules of the PBA. In some cases, the pharmaceutical composition can comprise one molecule of TBA and at least 3 molecules of the PBA. In some cases, the pharmaceutical composition can comprise one molecule of TBA and at least 4 molecules of the PBA. In some cases, the pharmaceutical composition can comprise one molecule of TBA and at least 5 molecules of the PBA. In some cases, the pharmaceutical composition can comprise one molecule of TBA and at least 6 molecules of the PBA. In some cases, the pharmaceutical composition can comprise one molecule of TBA and at least 8 molecules of the PBA.

[0230] The pharmaceutical composition disclosed herein can comprise a PBA to TBA ratio in a range of from 1 to 24. In some cases, pharmaceutical composition can comprise a TBA to PBA ratio in a range of from 1, 2, 4, 6, 8, 10, 12, 16, 20, or 24. When the TBA is selected from an antibody, the TBA to PBA ratio can also be referred to as drug-to- antibody ration (DAR). The pharmaceutical composition disclosed herein can comprise a Nano-ADC having a DAR in a range of from 1 to 24. The pharmaceutical composition can have a DAR of 1, 2, 4, 6, 8, 10, 12, 16, 20, or 24. In some cases, The pharmaceutical composition can have a DAR 2, 4, 6, 8, 10, 12, 16, 20, or 24. In some cases, the pharmaceutical composition can comprise two or more different PBAs and can have a total DAR of 1, 2, 4, 6, 8, 10, 12, 16, 20, or 24.

[0231] In some cases, the pharmaceutical composition can comprise nanoaggregate having a size less than 140 nm before lyophilization. In particular cases, the pharmaceutical composition can comprise nanoaggregate having in a range of from 1 to 140 nm, 1 to 120 nm, 1 to 100 nm, 1 to 80 nm, 1 to 60 nm, 1 to 40 nm, 1 to 20 nm, 1 to 15 nm, 1 to 10 nm, 1 to 8 nm, 1 to 6 nm, 2 to 120 nm, 4 to 120 nm, 4 to 100 nm, 4 to 80 nm, 4 to 60 nm, 4 to 40 nm, 4 to 30 nm, 4 to 20 nm, 4 to 15 nm, before lyophilization. In some cases, the pharmaceutical composition can comprise nanoaggregate having in a range of from 15 to 120 nm, 15 to 100 nm, 15 to 80 nm, 15 to 60 nm, 15 to 40 nm, 15 to 30 nm, 15 to 20 nm, 10 to 120 nm, 10 to 100 nm, 10 to 80 nm, 10 to 60 nm, 10 to 40 nm, 10 to 30 nm, 10 to 20 nm, 5 to 120 nm, 5 to 100 nm, 5 to 80 nm, 5 to 60 nm, 5 to 40 nm, 5 to 30 nm, 5 to 20 nm, 5 to 10 nm, 2 to 120 nm, 2 to 100 nm, 2 to 80 nm, 2 to 60 nm, 2 to 40 nm, 2 to 30 nm, 2 to 20 nm, 2 to 15 nm, 2 to 10 nm, and 2 to 5 nm, before lyophilization. The size of nanoaggregate can be measured using a dynamic light scattering (DLS) method. Other methods known to those skilled in the art can also be used.

[0232] In some cases, the polymer can comprise polyethylene glycol (PEG) having monomer units in a range of from 1 to 100, i.e. , m=1-100 in formulas disclosed herein. In some cases, the polymer can comprise polyethylene glycol (PEG) having monomer units in a range of from 12 to 36, 12 to 30, 12 to 24, 12 to 20, 12 to 18, 12 to 16, and 12 to 14. In some cases, the polymer can comprise polyethylene glycol (PEG) having monomer units in a range of from 20 to 36, 20 to 30, 20 to 26 and 20 to 24. In some cases, the polymer can comprise polyethylene glycol (PEG) having 24 monomer units.

[0233] In some cases, the pharmaceutical composition can comprise an antibody-drug conjugate (ADC), wherein the ADC comprises a formula selected from Formula ID 50, 51 , 52, 53, 54, 55, 56, 57, 58, 59, 60, 61 , 62, 63, 64, 65, and a combination thereof, as disclosed above.

[0234] It is understood by those skilled in the art that although n is an integer in a range of from 1 to 100, actual measurements of a TBA: PBA ratio can be a number in any of the aforementioned ranges including a fraction due to practical measurement limitations and / or the potential presence of some un-conjugated TBA, such as un-conjugated antibody. As disclosed above, the pharmaceutical composition can comprise a measured TBA:PBA in a range of from 1 :0.01 to 1 :100. In some cases, the pharmaceutical composition of this disclosure can comprise free antibody that is not conjugated with a PBA, as disclosed in detail hereafter. In some cases, the pharmaceutical composition ofthis disclosure can a Drug-to-Antibody Ratio (DAR), i.e. , PBA:TBA molar ratio, in a range of 1 :100 to 1 :0.01.

[0235] A symbol typically depicting an antibody is used in the formulas throughout this disclosure. Although a typical two-stranded antibody symbol is used in the drawings, an antibody can be a single chain, multi-chain, or a fragment of an immunoglobin (Ig), IgA, IgD, IgE, IgG, IgM, a part thereof, or a combination thereof. The term “antibody” used herein throughout this disclosure can refer to a typical two chain antibody having heavy chains and light chains arranged in three portions (or regions) in a Y shape, a two heavy chain antibody that lacks light chains, a single chain antibody, a multi-valent antibody that can comprise binding sites having affinities to 2 or more targeting sites or multiple antigen binding affinities, such as a bi-specific antibody, a tri-specific antibody, a tetra-specific antibody, a part thereof (i.e., a fragment of an antibody), an antigen-binding portion thereof or a combination thereof.

[0236] In some cases, the pharmaceutical composition can be a drug for treating cancer.

[0237] Although the term “mExa” is specifically used, any PBA disclosed herein, or known to those skilled in the art can be used.

[0238] In any one of the Nano-ADCs disclosed in figures or texts, n or n’ each is an integer that can be independently in a range of from 1 to 100. In some cases, the Nano-ADCs disclosed herein can have m=12, 18, 20, 24, 26, 28, and 30. In some cases, the Nano- ADCs disclosed herein can have m=24. In some cases, the Nano-ADCs disclosed herein can have n=1 -20. In some cases, the Nano-ADCs disclosed herein can have a measured n=1-9 (with a nominal n=1-8). In some cases, the Nano-ADCs disclosed herein can have n=8-9. In some cases, n is 4 that can produce a Nano-ADC having DAR = 4. In some cases, n is 8 that can produce a Nano-ADC having DAR = 8. In some cases, n is 16 that can produce a Nano-ADC having DAR = 18. In some cases, n+n’ can be in a range of from 2 to 100. In some cases, n+n’ can be in a range of from 2 to 24.

[0239] In some cases, a schematic illustration of a Nano-ADC having Formula ID 3001 is depicted in FIG. 10A comprising an antibody (14), a PEG polymer having 1 -100 units, i.e., m=1-100, a covalently linked, i.e., reacted, cleavable linker GGFG (Formula ID 135), and a modified exatecan having Formula ID 1 (FIG. 10A).

[0240] In some cases, a Nano-ADC having Formula ID 3002 is depicted in FIG. 10B comprising an antibody (14), a PEG polymer having 1 -100 units, i.e., m=1-100, a covalently linked, i.e., reacted, cleavable linker GGFG, and a modified exatecan having Formula ID 2 (FIG. 10B).

[0241] In some cases, a Nano-ADC having a Formula ID 3003 is depicted in FIG. 10C comprising an antibody (14), a PEG polymer having 1-100 units, i.e., m=1-100, a covalently linked, i.e., reacted, cleavable linker GGFG, and a modified exatecan having Formula ID 3 (FIG. 10C).

[0242] In some cases, a Nano-ADC having a Formula ID 3004 is depicted in FIG. 10D comprising an antibody (14), a PEG polymer having 1-100 units, i.e., m=1-100, a covalently linked, i.e., reacted, cleavable linker GGFG, and a modified exatecan having Formula ID 1006 (also referred to as EE) (FIG. 10D).

[0243] In some cases, a Nano-ADC having a Formula ID 3005 is depicted in FIG. 10E comprising an antibody (14), a PEG polymer having 1-100 units, i.e., m=1-100, a covalently linked, i.e., reacted, cleavable linker GGFG, and a modified exatecan having Formula ID 1029 (also referred to as CnE) (FIG. 10E).

[0244] In some cases, a Nano-ADC having a Formula 3006 is depicted in FIG. 10F comprising an antibody (14), a PEG polymer having 1-100 units, i.e., m=1-100, a covalently linked, i.e., reacted, cleavable linker GGFG, and a modified exatecan having a Formula ID 1012 (also referred to as BzE) (FIG. 10F).

[0245] In some cases, a Nano-ADC having a Formula ID 3007 is depicted in FIG. 10G comprising an antibody (14), a PEG polymer having 1-100 units, i.e., m=1-100, a covalently linked, i.e., reacted, cleavable linker GGFG, and a modified exatecan having Formula ID 1009 (also referred to as PE) (FIG. 10G).

[0246] In some cases, a Nano-ADC having a Formula ID 3008 is depicted in FIG. 10H comprising an antibody (14), a PEG polymer having 1-100 units, i.e., m=1-100, a covalently linked, i.e., reacted, cleavable linker GGFG, and a modified exatecan having a Formula ID 1008 (also referred to as CE) (FIG. 10H).

[0247] In some cases, a Nano-ADC having a Formula 3009 is depicted in FIG. 101 comprising an antibody (14), a PEG polymer having 1-100 units, i.e., m=1-100, a covalently linked, i.e., reacted, cleavable linker GGFG, and a modified exatecan having a Formula ID 1005 (also referred to as TFE) (FIG. 101).

[0248] In some cases, a Nano-ADC having a Formula ID 3010 is depicted in FIG. 10J comprising an antibody (14), a PEG polymer having 1-100 units, i.e., m=1-100, a covalently linked, i.e., reacted, cleavable linker GGFG, and a modified exatecan having a Formula ID 1007 (also referred to as I PE) (FIG. 10J).

[0249] In some cases, a Nano-ADC having a Formula ID 3010 is depicted in FIG. 10K comprising an antibody (14), a covalently linked, i.e., reacted cleavable linker GGFG, anda modified exatecan having a Formula ID 1006 (also referred to as EE) (FIG. 10K). In some cases, a Nano-ADC can be free from a polymer disclosed herein.

[0250] In some cases, a Nano-ADC can comprise a reacted compound selected from Formula ID 1 , 1-1 , 1-2, 2, 2-1 , 2-2, 3, 3-1 , 3-2, 1001 through 1135 and a combination thereof, and a covalently linked, i.e., reacted cleavable linker, selected from Formula ID 101-141 , and a combination thereof. In some cases, a Nano-ADC can comprise a reacted GGFG or any other cleavable linkers disclosed herein, without a PEG polymer, similar to Formula ID 3011 depicted in FIG. 10K.

[0251] In some cases, a Nano-ADC can comprise any one of the cleavable linkers disclosed herein or a combination thereof.

[0252] Further examples of Nano-ADCs of this disclosure are shown in FIG. 11A - FIG. 11 F and FIG. 12A-FIG. 12Q. In some cases, representative examples of Nano-ADCs can comprise Formula ID 3012, 3013, 3014, or a combination thereon, that each can comprise an antibody (14), one or more GGFG cleavable linkers and at least a mExa (FIG. 11A-FIG. 11C). In some cases, representative examples of Nano-ADCs can comprise Formula ID 3015, 3016, 3017, or a combination thereof, that each can comprise an antibody (14), one or more GGFG cleavable linkers and multiple mExa, such as two or more mExa (FIG. 11 D-FIG. 11 F). When applicable, m is an integer that can be in a range of from 1 to 100. n is an integer that can be in a range of from 1 to 100.

[0253] In some cases, representative examples of Nano-ADCs can comprise Formula ID 3018, 3019, 3020, or a combination thereof, that each can comprise an antibody (14), one or more VAPAB or GGFG cleavable linkers and at least a mExa (FIG. 12A-FIG. 12C). In some cases, representative examples of Nano-ADCs can comprise Formula ID 3021 , 3022, 3023, or a combination thereof, that each can comprise an antibody (14), one or more VAPAB cleavable linkers and multiple mExa, such as two or more mExa (FIG. 12D-FIG. 12F). In some cases, a Nano-ADC can comprise Formula ID 3024 that can comprise an antibody (14), a PEG24, a GGFG linker and a modified exatecan (FIG. 12G) In some cases, a Nano-ADC can comprise Formula ID 3025 that can comprise an antibody, such as an anti-HER2 monoclonal antibody trastuzumab biosimilar (KANJINTI®), a modified exatecan, a GGFG linker and without any polymer (FIG. 12H). In some cases, a Nano-ADC can comprise a Formula ID 3026 that can comprise an antibody (14), a PEG24, a VAPAB linker and a modified exatecan (EE) (FIG. 121). In some cases, a Nano-ADC can comprise a Formula ID 3027 that can comprise an antibody (14), a polymer side chain comprising a PEOX polymer linked to a lysine residue and an unmodified exatecan (E) (FIG. 12J). In some cases, a Nano-ADC can comprise a FormulaID 3028 that can comprise the Kan antibody, a PEG24, a VAPAB linker and a modified exatecan (EE) with DAR in a range of from 4 to 8 (n = 4-8) (FIG. 12K). In some cases, a Nano-ADC can comprise a Formula ID 3029 that can comprise the Kan antibody, a PEG24, a GGFG linker and a modified exatecan (TFE) (Fig. 12L). In some cases, a Nano- ADC can comprise a Formula ID 3030 that can comprise the Kan antibody, a polymer side chain comprising a PEOX polymer linked to a lysine residue, a GGFG linker and an un-exatecan (E) (FIG. 12M). In some cases, a Nano-ADC can comprise a Formula ID 3031 that can comprise a B7H3 antibody IFI, a polymer side chain comprising a PEOX polymer linked to a lysine residue, and an un-modified exatecan (E) (FIG. 12N). In some cases, a Nano-ADC can comprise a Formula ID 3032 that can comprise an IFI antibody, a polymer side chain comprising a PEOX polymer linked to a lysine residue, such as PEOX10, and an un-modified exatecan (E) (FIG. 120). In some cases, a Nano-ADC can comprise a Formula ID 3033 that can comprise an B7H3 antibody IFI, a polymer side chain comprising a PEOX polymer linked to a lysine residue of a first polymer, and an unmodified exatecan (E), and a second polymer comprise a PEG, and a MMAE (FIG. 12P), wherein the first polymer and the second polymer are covalently linked to the B7H3 antibody, such as via cystine-based conjugations. In some cases, a Nano-ADC can comprise a Formula ID 3034 that can comprise an antibody (14), a polymer side chain comprising a PEOX polymer linked to a lysine residue of a first polymer, and an unmodified exatecan (E), and a second polymer comprise a PEG, and MMAE (FIG. 12Q), wherein the first polymer and the second polymer are covalently linked to the antibody, such as via cystine-based conjugations. In some cases, a Nano-ADC can comprise a Formula ID 3035 that can comprise an antibody (14), a polymer side chain comprising a PEOX polymer linked to a lysine residue of a first polymer, and an un-modified exatecan (E), and a second polymer comprise a PEG, and a modified exatecan TFE (Formula ID 1005) (FIG. 12R), wherein the first polymer and the second polymer are covalently linked to the antibody, such as via cystine-based conjugations. In some cases, the antibody in Formula ID 3035 can be ifinatamab (IFI). Formula ID 3036 Kan[-(MC-L(PEO)-p-E)n, - (Mal-PEG2-VCP-MMAE)n’] can be produced by conjugating trastuzumab biosimilar (KANJINTI®, Kan) antibody with mixed biomolecules -SMCC-L(PEOX10)-p-E and -Mal- PEG2-VC-MMAE, with y=12, 18, 24 (FIG. 12S). In some cases, a Nano-ADC can have Formula ID 3037 with the Kan antibody, PEOX polymer side chain having y=10, a VP linker and an un-modified exatecan, Kan(-SMCC-L(PEOX10)-VP-E)n. In some cases, Formula ID 3037 can have n=8, i.e., DAR=8 (FIG. 12T). In some cases, Formula ID 3038 can have y=18, n=8, i.e., DAR=8 (FIG. 12U). In some cases, a Nano-ADC can be freefrom a para-aminobenzyl group and can provide optimized cleavage in plasma v.s. in cells, such as in the tumor cells. A Nano-ADC that is free from the para-aminobenzyl group can have better stability in plasma reducing undesired cleavage on the ADC before the drug reaches the target, such as tumor cells. When applicable, m is an integer that can be in a range of from 1 to 100, y is an integer in a range of from 1 to 100, n is an integer that can be in a range of from 1 to 100. In some cases, n can be 4. In some cases, n can be 8. In some cases, n can be 16. In some cases, n+n’ can be 4. In some cases, n+n’ can be 8. In some cases, n+n’ can be 16. In some cases, n+n’ can be 24. In some cases, n:n’ can be in a range of from 1 :10 to 10:1 . In some cases, an Nano-ADC can have the Formula ID 3035 with n:n’ in a range of from 1 :8 to 8:1. In some cases, an Nano-ADC can have the Formula ID 3036 with n:n’ in a range of from 1 :8 to 8:1. In some cases, an Nano-ADC can have n’:n = 2:1 to 8:1. In some cases, an Nano-ADC can have n’:n = 1 :2 to 1 :8. In some cases, an Nano-ADC can have n’:n = 3:1 to 8: 1 . In some cases, an Nano- ADC can have n’:n = 1 :3 to 1 :8.

[0254] In some cases, the pharmaceutical composition disclosed herein can comprise a Nano-ADC comprising multiple polymers and multiple PBAs. In some cases, the pharmaceutical composition disclosed herein can comprise a Nano-ADC comprising at least two polymers and at least one of the polymers can be linked to a PBA. In some cases, the pharmaceutical composition disclosed herein can comprise a Nano-ADC comprising at least two polymers and each of the polymers can be linked to at least a same or different PBA. In some cases, at least one of the at least two polymers can comprise PEOX polymer a polymer side chain that comprises a PEOX polymer, or a combination thereof. In some cases, the pharmaceutical composition disclosed herein can comprise a Nano-ADC comprising a first polymer polymers and a second polymer, wherein the first polymer is linked to a first PBA (herein “PBA1 ”) and the second polymer is linked to a second PBA (herein “PBA2”), wherein the first polymer and the second polymer can be the same or different and the first PBA and the second PBA can be the same or different. In some cases, the first polymer and the second polymer can be linked to a same antibody at different sites. In some cases, the first polymer and the second polymer can be linked to a same antibody at different cystine residues. In some cases, the pharmaceutical composition disclosed herein can comprise a Nano-ADC Formula ID 3033, 3034, 3035, 3036, 3044, 3066, 3067, 3068 3069 or a combination thereof.

[0255] In some cases, the pharmaceutical composition disclosed herein can comprise a Nano-ADC comprising a first polymer polymers and a second polymer, wherein the first polymer is linked to a first PBA and the second polymer is linked to a second PBA, whereinthe first polymer, the second polymer, or a combination thereof can comprise a PEOX polymer, a polymer side chain comprising a PEOX polymer, or a combination thereof, and wherein at least one of the first PBA, the second PBA or a combination thereof, comprises a Formula ID 1001-1135, calicheamicin, DM1 (antimicrotubule agent T-DM1 , a derivative of maytansine), DM4 (Ravtansine), Dxd (deruxtecan), exatecan, IRDye700DX (a nearinfrared (NIR) photosensitizer suitable for near infrared (NIR) photoimmunotherapy (PIT)), MMAE (Monomethyl auristatin E, also known as vedotin), MMAF (Monomethyl auristatin F), pseudomonas exotoxin A (a potent exotoxin produced by the bacterium Pseudomonas aeruginosa), pyrrolobenzodiazepine (PBD), SG3199 (a cytotoxic DNA minor groove interstrand crosslinking pyrrolobenzodiazepine (PBD) dimer), SN-38, irinotecan, camptothecin, amanitin (an inhibitor of RNA Polymerase II), duocarmycins (DNA- alkylators), halichondrin (Eribulin, also known Halaven®, under respective trademark), norhalichondrin B, maytansinoids, maitansine (INN), maytansine, PUN-159682 (an inhibitor of DNA Topoisomerase II), tubulysins, tubulysin, pretubulysin (PT), tubulysin A (TubA), tubulysin M, a pharmaceutically acceptable salt thereof, a solvate thereof, a prodrug thereof, an isomer thereof, an isotope derivative thereof, and a combination thereof.

[0256] In some cases, the pharmaceutical composition disclosed herein can comprise a Nano-ADC listed in Table 1 , or a combination thereof.Table 1. Representative examples of Nano-ADC(Each formula is in a reacted form in a Nano-ADC).Note: “Kan” refers to an anti-HER2 monoclonal antibody trastuzumab Herceptin® biosimilar (KANJINTI®). “B7H3” refers to a monoclonal antibody against the Human B7 homolog 3 (B7-H3) protein that is a member of the B7 family of immune proteins. “IFI” refers to ifinatamab anti-human B7-H3 (also referred to as “B7H3”) (CD276) monoclonal antibody. “HlgG” refers to non-specific human IgG that is used as a comparative. The structures are reacted to the antibody via conjugations (herein “conjugated” or “reacted”) with n and n’ as defined herein.

[0257] Although specific S- bond and N- bond are depicted in figures and descriptions, all aforementioned conjugations and or bonds can be suitable. In some case, thepharmaceutical composition can comprise a Nano-ADC conjugated via any of the aforementioned bonds or conjugates, such as S- bond, N-bond, glycol bond, modified glycol bond, GIcNAc (N-acetylglucosamine), substituted GIcNAc, GIcNAc-galactose, GIcNAc-mannose, GIcNAc-glucose, GIcNAc- glucuronic acid, GIcNAc-fucose, GIcNAc-N- acetylneuraminic acid, site specific bond, random bond, bioconjugations to other amino acid residues of the antibody, such as tyrosine (Tyr), tryptophan (Trp), and methionine (Met), conjugations via genetically engineered non-natural amino acids (NNAA), enzymatic conjugations, or a combination thereof.

[0258] In some cases, the pharmaceutical composition can comprise two or more same or different PBAs. In some cases, the bioactive composition can comprise Formula ID 3033, 3034, 3035, 3036, 3044 (comprising a conjugated (also referred to as reacted) Formula ID 2005), 3066 (comprising a conjugated Formula ID 2027), 3067 (comprising a conjugated Formula ID 2028), 3068 (comprising a conjugated (reacted Formula ID 2029), 3069 (comprising a conjugated Formula ID 2030), 3076 (comprising a conjugated Formula ID 2037) or a combination thereof.

[0259] In some cases, the pharmaceutical composition can comprise a TBA, i.e., an antibody, one or more PBAs, at least one linker, and a polymer side chain attached to the linker, such as Formula ID 2025, 2026, 2027 or a combination thereof.

[0260] Although the specific locations of linkers, cleavable linkers or specific peptide linkers are illustrated in figures and descriptions as examples, any of the linkers disclosed herein can be positioned at L1 , L2, or a combination thereof. In some cases, the Nano- ADC can comprise a linker positioned between the antibody and the polymer. In some cases, the Nano-ADC can comprise a linker positioned between the polymer and the PBA. In some cases, the Nano-ADC can comprise a linker positioned between the antibody and the polymer and a linker positioned between the polymer and the PBA. The linkers can be the same or different. The term “Linker”, “Linker L1 ,” “L1 Linker,” “L1 ,” “Linker L2,” “L2 Linker,” “L2,” “cleavable Linker,” or a gametic variation, regardless of whether capitalized or not, refers to a linker either in its un-reacted form or a reacted form, i.e., incorporated in an ADC.

[0261] In some cases, a pharmaceutical composition can comprise an antibody-drug conjugate (ADC), wherein the TBA can be an antibody (14), such as an anti-HER2 antibody. In some cases, an antibody-drug conjugate (Nano-ADC) can comprise an antibody, a polymer, and at least a linker. In some case, a pharmaceutical composition comprises a Nano Antibody-drug Conjugate (Nano ADC) comprising a payload bioactive agent (PBA) comprising a formula selected from:a pharmaceutically acceptable salt thereof, a solvate thereof, a prodrug thereof, an isomer thereof, an isotope derivative thereof, a reacted PBA thereof, optionally an additional cytotoxic agent, and a combination thereof; and an antibody selected from a monoclonal antibody (mAb) HER2 receptor antagonist trastuzumab biosimilar, an anti-human B7-H3 (CD276) monoclonal antibody, or a nonspecific human IgG (HlgG); optionally, a pharmaceutical suitable carrier; wherein, v and w each is an integer, v=1-100; w=1 -1000;R, R’, R”, R3, R4, R3’, R4’, Rs, Re, R7, Rs, and R15 each is independently H, D, CI- 0100 saturated or unsaturated hydrocarbon, C1-C100 saturated or unsaturated hydrocarbon having at least one D, C1-C100 saturated or unsaturated hydrocarbonhaving at least one heteroatom, C1-C100 saturated or unsaturated hydrocarbon having at least one heteroatom and at least one D, aromatic group, substituted aromatic group, or a combination thereof;R9 is selected from H, D, C1-C100 saturated or unsaturated hydrocarbon, C1-C100 saturated or unsaturated hydrocarbon having at least one D, C1-C100 saturated or unsaturated hydrocarbon having at least one heteroatom, C1-C100 saturated or unsaturated hydrocarbon having at least one heteroatom and at least one D, OH, NH2, Cl, F, I, or Br;R10 is selected from H, Cl, F, I, Br, and methoxy;R11 is OH, OR12, SH, SR13, NH2, NHD, ND2, N RUR’ , wherein R12 is an alcohol protecting group, R13 is a thiol protecting group, and Ru and R’14 each is independently H, D, or an amine protecting group; said heteroatom is selected from N, O, S, P, Cl, Br, I and a combination thereof; and X is O, S, or NH.

[0262] The polymer can comprise polyethyleneglycol (PEG), poly(2-oxazoline), poly(2- substituted oxazoline) that comprises poly(2-methyloxazoline), poly(2-ethyloxazoline), poly(2-propyloxazoline), poly(2-isopropyloxazoline) (PiPOX), poly(2-substituted oxazoline), or a combination thereof. The POX can comprise poly(2-methyloxazoline) (PMOX) in one example, poly(2-ethyloxazoline) (PEOX) in another example, poly(2- propyloxazoline) (PPOX) in yet another example, poly(2-isopropyloxazoline) (PiPOX) in yet another example, or a combination of two or more of the poly(2-substituted oxazoline)s in yet a further example, wherein the two or more of the poly(2-substituted oxazoline)s can be a repeating unit, also referred to as complex monomer, in the polyoxazoline polymer. The polyoxazoline (POX) is hydrophilic. The polyoxazoline (POX) can be free from monomers, either simple or complex monomers, having hydrophobic side chains, such as those having 4 or more carbons (C4 and above).

[0263] The modified SBP's can be obtained, for example, through chemically linking functional groups on, for example, symmetrically branched PAMAM or PPI dendrimers, commercially available from Aldrich, polyether dendrimers, polyester dendrimers, comb- branched / star-branched polymers, such as, those containing PEO, PEG, PMOX or PEOX, polystyrene, and comb-branched dendrigrafts, such as, those containing PEOX, PMOX or PEI . The synthetic procedures for making such SBP's / dendrimers are described above and hereafter.

[0264] In some cases, the higher branching densities of SBP's can render the polymers molecularly compact with a well-defined interior void space, which makes such molecules suitable as a carrier for water insoluble or poorly water soluble drugs.

[0265] The antibodies disclosed herein, or a combination thereof can be suitable as a TBA for the Nano-ADC of this disclosure. In addition, in some cases, the TBA can be selected from ABT-700, ABT-806, Anti-AGS16 (Hu lgG2a), Anti-CD19, Anti-CD22, Anti-CD22 (Hz lgG1 ), Anti-CD79b (Hz lgG1 ), Anti-DLL3 (Rovalpituzumab), humanized anti-DLL3 DB131401 , Anti-LIV1 (Hz lgG1), Antimesothelin (lgG1), Anti-PSMA (Hu lgG1 ), cAC10 (SGN-30, Ch-lgG1), CR-011 (Hu lgG2), DS6 (Hu lgG1 ), Enfortumab, G5 / 44 (Hz lgG4), Hp67.6 (Hz lgG4), hRS7 IgGk, huB4 (Hz lgG1 ), huN901 (Hz lgG1 ), lgG1 , K7153A humanized lgG1 , M9346A, Nbt062, Anti- CD138 (Ch lgG4), Tisotumab, Trastuzumab (Herceptin), Trastuzumab (Hz lgG1), and a combination thereof.

[0266] In some cases, the TBA can have binding affinity to DLL3, gpNMB, PSMA, CD22, ABT-700, LIV-1 , FOLR1 , CD56, CD19, CD138, Mesothelin, CA6, EGFR, CD37, ENPP3, CD33, CD30, HER2, CD79b, Nectin4, Trop-2, BCMA, Tissue factor, or a combination thereof.

[0267] In some cases, the Nano-ADC of this disclosure can comprise additional PBA that can comprise a natural or synthetic small molecule-based drug, inorganic-based drug, a toxin molecule having cytotoxicity, a molecule having cytotoxicity to the target cell, a derivative thereof, or a combination thereof. In some cases, the PBA can comprise 5- Fluorouracil (5-Fll), alkylators, anti-metabolites, cabazitaxel, Calicheamicin, Calicheamicin derivatives, camptothecin, camptothecin derivatives, capecitabine, cell division inhibitors or microtubule inhibitors, ci profl oxaxi n, cyclophosphamide, deruxtecan, deruxtecan derivatives, DNA cleavage agent, DNA crosslinking agent, DNA replicating inhibitor, docetaxel, dolastatin analogs, doxorubicin, duocarmycin analogs, etoposide, everolimus, exatecan, exatecan derivatives, topoisomerase inhibitors, gemcitabine, irinotecan (CPT-11 ), larotaxel, maytansinoids (DM1 , DM4), milataxel, MMAE (monomethyl auristatin E), MMAF (monomethyl auristatin F), MMAH (monomethyl auristatin H), mTOR inhibitor, one or more ATP-competitive mTORC1 / 2 inhibitors, one or more dual PI3K- mTOR inhibitors, ortataxel, ozogamicin, paclitaxel, pemetrexed, pyrrolobenzodiazepine dimer (PBD), rapamycin, ridaforolimus, SN-38, taxane, temsirolimus, tesetaxel, Topoisomerase 1 (Top 1 ) inhibitor, Topoisomerase 2 (Top 2) inhibitor, topotecan, torin-1 , torin-2, vinblastine, vinca alkaloids, vincristine, vinorelbine, vistusertib, zotarolimus, biological toxins, ricin, or a combination thereof.

[0268] The terms “Linker,” “Linker 1 ,” or “Linker 2” represents any linkers disclosed herein and is used for illustration purposes only. The Linker, Linker 1 (also shown as L1 ) and Linker 2 (also shown as L2) can be the same or different or in a combination thereof.

[0269] In some cases, n can be an integer in a range of from 1 to 100. In some cases, the pharmaceutical composition disclosed above can comprise a molar ratio of antibody to the drug, i.e., TBA:PBA, in a range of 1 :1 to 1 :100, 1 :2 to 1 :100, 1 :4 to 1 :100, 1 :8 to 1 :100, 1 :12 to 1 :100, 1 :20 to 1 : 100, 1 :24 to 1 :100, 1 :48 to 1 :100 and 1 :64 to 1 :100. In some cases, the pharmaceutical composition disclosed above can comprise a molar ratio of antibody to the drug, i.e., TBA:PBA, in a range of from 1 :2 to 1 :24. In some cases, the pharmaceutical composition disclosed above can comprise a molar ratio of antibody to the drug, i.e., TBA:PBA, in a range of from 1 :4 to 1 :24. In some cases, the pharmaceutical composition disclosed above can comprise a molar ratio of antibody to the drug, i.e., TBA:PBA, in a range of from 1 :8 to 1 :24. In some cases, the pharmaceutical composition disclosed above can comprise a molar ratio of antibody to the drug, i.e., TBA:PBA, in a range of from 1 :16 to 1 :24. One advantage of the ADC disclosed herein is that the number of the drug attached to each antibody can be more than two in one example, more than four in another example, more than 8 in yet another example, more than 10 in yet another example, more than 12 in yet another example, more than 16 in yet another example, and more than 24 in yet another example. It is understood by those skilled in the art that although n is an integer in a range of from 1 to 100, actual measurements of a TBA:PBA ratio can be a number in any of the aforementioned ranges including a fraction due to practical measurement limitations and the potential presence of some un-conjugated TBA, such as un-conjugated antibody.

[0270] In some cases, the pharmaceutical composition disclosed herein can comprise two or more of the PBAs disclosed herein. In some cases, the pharmaceutical composition can comprise two or more PBAs that can comprise Formula ID 1 , 1-1 , 1-2, 2, 2-1 , 2-2, 3, 3-1 , 3-2, 1001-1135, exatecan, deruxtecan, MMAE, MMAF, a pharmaceutically acceptable salt thereof, a solvate thereof, a prodrug thereof, an isomer thereof, an isotope derivative thereof, or a combination thereof. In some cases, the pharmaceutical composition can comprise two or more PBAs that can comprise a first PBA and s second PBA, wherein the first PBA and the second PBA can be the same or different. In some cases, the first PBA and the second PBA can be independently selected from Formula ID 1 , 1-1 , 1 -2, 2, 2-1 , 2-2, 3, 3-1 , 3-2, 1001-1135, exatecan, deruxtecan, MMAE, MMAF, a pharmaceutically acceptable salt thereof, a solvate thereof, a prodrug thereof, an isomer thereof, an isotope derivative thereof, or a combination thereof.

[0271] In some cases, the first PBA can be linked to one or more of the second terminal groups of the polymer. In some cases, the second PBA can be linked to one or more second terminal groups of the polymer side chain or multiple polymer side chains. In some cases, the two or more same or different first PBAs can be linked to two or more of the second terminal groups of the polymer. In some cases, the two or more same or different second PBAs can be linked to two or more second terminal groups of the polymer side chains.

[0272] In some cases, the first and the second PBAs can comprise two or more same PBAs. In some cases, the first and the second PBAs can comprise two or more different PBAs.

[0273] In some cases, the pharmaceutical composition can comprise a mixture of TBA, wherein some of the TBA can be covalently linked to the PBA and some of the TBA can be non-covalently mixed with the PBA leading to more TBAs than the PBAs. In some cases, some antibody molecules can be free from bound drugs, i.e., free antibody without conjugated to a drug molecule. In some cases, the molar ratio of TBA:PBA can be 100: 1 , 20: 1 , 10:1 , 5:1 , 4;1 , 3;1 , or 2:1 (on a reverse ratio of 1 :0.01 , 1 :0.1 , 1 :0.2, 1 :0.25, 1 :0.3, and 1 :0.5). In some cases, the molar ratio of antibody to a drug can be 100: 1 , 10:1 , 5:1 , 4; 1 , 3; 1 , or 2:1 (on a reverse ratio of 1 :0.01 , 1 :0.1 , 1 :0.2, 1 :0.25, 1 :0.3, and 1 :0.5).

[0274] The terms “cancer” or “cancers” used herein and throughout this disclosure refers to cancer or tumor and can include malignant tumors and benign tumors such as solid tumors and cancers of the blood, such as leukemias. Malignant tumors can spread into, or invade, nearby tissues. In addition, as these tumors grow, some cancer cells can break off and travel to distant places in the body through the blood or the lymph system and form new tumors (metastatic tumor) far from the original tumor (primary cancer). A cancer can include a primary cancer or a metastatic tumor. The pharmaceutical composition disclosed herein can be a cancer treatment drug for treating one or more cancers. In some cases, the terms “cancer” or “cancers” used herein can include one or more cancer selected from acoustic neuroma, Acute Lymphoblastic Leukemia (adult), Acute Lymphoblastic Leukemia (pediatric), Acute Myeloid Leukemia, adenocarcinoma, Anal Cancer, Anemia and Neutropenia (Low Red and White Blood Cell Counts), basal cell carcinoma, Basal Cell Skin Cancer, B-Cell Lymphomas (Diffuse Large B-Cell Lymphoma), B-Cell Lymphomas (Follicular Lymphoma), B-Cell Lymphomas (Mantle Cell Lymphoma), bile duct carcinoma, biliary track cancer, Bladder Cancer, bladder carcinoma, Bone Cancer, Brain Cancer (Gliomas), brainstem glioma, breast cancer, Breast Cancer (DCIS Breast Cancer), Breast Cancer (Invasive Breast Cancer), BreastCancer (Metastatic Breast Cancer), triple-negative breast cancer, ER(+) locally advanced or metastatic breast cancer, bronchogenic carcinoma, Central Nervous System Cancers (Primary Central Nervous System Lymphoma), cervical cancer, choriocarcinoma, Chronic Lymphocytic Leukemia, chronic lymphocytic lymphoma, Chronic Myeloid Leukemia, colon cancer, colon carcinoma, Colorectal Cancer (CRC), diffuse large B cell lymphoma, ependymoma, Esophageal Cancer, Gallbladder and Bile Duct Cancers, gastric cancers, germinoma, glioblastoma astrocytoma, mixed gliomas, Graft-Versus-Host Disease, head and neck cancers, Head and Neck Cancers (Nasopharyngeal Cancer), Head and Neck Cancers (Oral Cancers), Head and Neck Cancers (Oropharyngeal Cancer), hemangioblastoma, hepatocellular carcinoma, hepatoma, Hodgkin Lymphoma, Kidney Cancer, leukemias, Liver Cancer, Lung Cancer, Lung Cancer (Non-Small Cell Lung Cancer - Early and Locally Advanced), Lung Cancer (Non-Small Cell Lung Cancer - Metastatic), Lung Cancer NSCLC (Non-Small Cell Lung Cancer), Lung Cancer (Small Cell Lung Cancer), lymphoid malignancy, Malignant Pleural Mesothelioma, medullary carcinoma, medullary thyroid carcinoma, medulloblastoma, melanoma, meningioma, Multiple Myeloma, Mycosis Fungoides / Sezary Syndrome, Myelodysplastic Syndromes, Myeloproliferative Neoplasms, neuroblastoma, neuroendocine tumor (advanced), Neuroendocrine Tumors, oligodendroglioma, ovarian cancer, pancreatic cancer, papillary adenocarcinomas, papillary carcinoma, papillary thyroid carcinoma, Peripheral T-Cell Lymphoma, perivascular epithelioid cell tumor (PEComa), perivascular epithelioid cell tumor (advanced unresectable or metastatic malignant), pheochromocytomas, pinealoma, Primary Cutaneous Lymphomas, prostate cancer, Prostate Cancer (Advanced Stage), Prostate Cancer (Early Stage), Rectal Cancer, recurrent endometrial cancer, relapsed ER(+) high-grade ovarian cancer, relapsed or refractory non-Hodgkin lymphoma, renal cancer (metastatic clear cell), renal cell carcinoma, retinoblastoma and brain metastases, Schwannoma craniopharyogioma, sebaceous gland carcinoma, seminoma, soft tissue sarcoma, squamous cell carcinoma, Squamous Cell Skin Cancer, Stomach Cancer, sweat gland carcinoma, Systemic Mastocytosis, testicular tumor, thyroid cancer, Uterine Cancer, uterine sarcoma, Waldenstrom Macroglobulinemia, and a combination thereof.

[0275] The term “a combination thereof” used for a combination of the bioactive agents disclosed above means a combination of two or more bioactive agents, wherein such combination does not have undesired effect, such as an undesired interaction between or among the bioactive agents. It is understood that some of combinations of the bioactive agents may not be suitable, or may not be desirable, such as those having undesiredinteractions. For example, a combination of theophylline and ciprofloxacin or warfarin and diflunisal may not be suitable. These combinations or any combinations determined by appropriate guidelines or regulations as not suitable are thus excluded.

[0276] In any of the pharmaceutical compositions disclosed above and hereafter, the nanoaggregate can have a weight ratio of polymer to bioactive agent in a range of from about 1 :500 to about 500:1. The nanoaggregate can have a weight ratio of the polymer to the bioactive agent in a range of from 1 :500 to about 500:1 , 1 :200 to 200:1 , 1 :100 to 200:1 , 1 :10 to 200:1 , 1 :5 to 200:1 , 1 : 1 to 200:1 , about 2:1 to about 200:1 in one example, about 2:1 to about 150:1 in another example, about 2:1 to about 120:1 in yet another example, about 2:1 to about 100:1 in yet another example, about 2:1 to about 80:1 in yet another example, about 2:1 to about 60:1 in yet another example, about 2:1 to about 40:1 in yet another example, about 2:1 to about 30:1 in yet another example, about 2:1 to about 20: 1 in yet another example, about 2:1 to about 15:1 in another example, about 2:1 to about 10:1 in yet another example, about 2:1 to about 8: 1 in yet another example, about 5:1 to about 10:1 in yet another example, about 5:1 to about 8:1 in yet another example, 5:1 in yet a further example, about 6: 1 to about 8: 1 in yet another example, 6:1 in yet a further example, 7:1 in yet a further example, 7.5:1 in yet a further example, and 8:1 in yet a further example. When pharmaceutical composition comprises two or more bioactive agents, the ratio of polymer to bioactive agent can be based on the total weight of polymer and the bioactive agents.

[0277] The pharmaceutical composition disclosed herein can be formulated for parenteral, oral, nasal, transdermal (topical), transmucosal, rectal administration, or a combination thereof and can comprise one or more pharmaceutical suitable carriers. In some cases, the pharmaceutical composition disclosed herein can be formulated for intravenous (IV), intradermal (ID), subcutaneous (SC), oral, transdermal (topical), transmucosal, rectal administration, or a combination thereof. In some cases, the pharmaceutical composition disclosed herein can be formulated for intravenous (IV), intradermal (ID), subcutaneous (SC), transdermal (topical) or transmucosal administration. In some cases, the pharmaceutical composition disclosed herein can be formulated for oral administration, such as tablets, capsules, oral spray, solutions, or suspensions. In some cases, the pharmaceutical composition disclosed herein can be formulated for nasal administration, such as nasal spray. The pharmaceutical suitable carriers disclosed herein can be suitable.

[0278] In some cases, this disclosure is directed to a method for treating or preventing a disease of a subject in need thereof, the method comprising administering the subject with an effective dose of the pharmaceutical composition disclosed herein.

[0279] Suitable to the method, the pharmaceutical composition can be administered to the subject via intravenous (IV) injection, subcutaneous (SC) injection, intramuscular (IM) injection, intradermal (ID) injection, oral administration, aerosol administration, or a combination thereof.

[0280] The method disclosed herein can further comprising the step of administering the subject with one or more subsequent bioactive agents selected from a protein, a peptide, an antibody, a fragment of an antibody, a chemical compound, a small molecule drug, one or more chemotherapy drugs, a vaccine, and a combination thereof, wherein each of the one or more subsequent bioactive agents is administered to the subject, prior to, at the same time as, or after administering the pharmaceutical composition.

[0281] The disease can be cancer.

[0282] The subject can be a human patient having a heterogenous tumor. The subject can comprise AgLowtumor cells that are adjacent to AgHightumor cells, distal to any AgHightumor cells, free from contact with any the AgHi9htumor cells, or a combination thereof.

[0283] Suitable to the method disclosed herein, the pharmaceutical composition can comprise a TBA comprising an antibody or a part thereof having binding affinity to HER2, AgHightumor cytotoxicity to HER2+tumor cells, and AgLowtumor cytotoxicity to HER2' (also known as HER2Low) or HER2 ultra-low tumor cells. The HER2' tumor cells can be adjacent to HER2+tumor cells, distal to any HER2+tumor cells, free from contact with any HER2+tumor cells, or a combination thereof. The subject can be a human patient having heterogenous HER2+and HER2' breast cancer, metastatic breast cancer, triple-negative breast cancer (TNBC), gastric cancer, colon cancer, or a combination thereof. In some cases, the subject can be a human patient having a cancer with low or no (i.e. , negative) tumor antigens or tumor markers. In some cases, the subject can be a human patient having a triple-negative breast cancer (TNBC) (HER2-).

[0284] In some cases, the Nano-ADC disclosed herein can comprise a tumor inhibition activity to inhibit tumor growth after a treatment with another tumor therapy. In some cases, the Nano-ADC disclosed herein can comprise a tumor inhibition activity to inhibit tumor growth together with a treatment with second tumor therapy, wherein the second tumor therapy alone fails to inhibit the tumor growth. In some cases, the Nano-ADC disclosed herein can comprise a tumor inhibition activity to inhibit tumor growth of a tumor having low tumor antigen (AgLow). In some cases, the Nano-ADC can comprise a tumorinhibition activity to inhibit tumor growth after failed treatment with an ADC famtrastuzumab deruxtecan-nxki (ENHERTU®) having a monoclonal antibody trastuzumab and deruxtecan. In some cases, the Nano-ADC can comprise a tumor inhibition activity to inhibit tumor growth together with ENHERTU®, wherein the ENHERTU® alone fails to inhibit the tumor growth. In some cases, the Nano-ADC disclosed herein can comprise a tumor inhibition activity to inhibit tumor growth of a tumor having low tumor antigen (AgLow), such as tumors having low HER2 antigen levels (HER2_or HER2Low).

[0285] Not wishing to be bound by a particular mechanism or theory, Applicants believe that, in addition to improve water solubility of water insoluble payload (PBA) drugs, the nanocomposition of this disclosure including the ADC produced herein, referred to as “Nano-ADC” herein, provides hydrodynamic sizes that can be an important factor in the specificity of drug delivery, herein referred to as “nanoparticle-mediated cytotoxicity (NMC)” to tumor cells. A potential model of this mechanism is schematically illustrated in FIG. 13A-FIG. 13C. It has been reported that accumulation of IV-injected nanoparticles in cancer tissues depends on the passive diffusion of these particles across the hyperpermeable tumor vasculature and retention in the interstitium via a slower clearance rate due to impaired lymphatics. This enables the encapsulated drug molecules to persist in situ for a longer time compared to non-encapsulated drugs. The basic mechanism for the specificity of drug delivery using nanoparticles can depend on the abnormality of blood vessels associated with tumors. Tumors typically contain abnormal vascularization wherein the blood vessels, unlike normal ones, have a discontinuous endothelium, with gaps between cells that may be several hundred nanometers large, creating sites of hyperpermeability in the tumor and allowing macromolecular transport to occur via these gaps. The effect can lower the non-specific toxicity of the drug since its size reduces its ability to enter normal cells, while allowing the drug to accumulate in neoplastic tissue due to their hyperpermeable vasculature. So far, no such nanoparticle-mediated cytotoxicity has been reported with ADCs. The Nano-ADCs disclosed herein are configured to be within a size range that can be suitable for nanoparticle-mediated cytotoxicity to tumor cells. The Nano-ADCs disclosed herein can also comprise cleavable linkers that can deliver the payload cytotoxic drug once the Nano-ADC enters tumor cells. The TBA antibody can help to deliver the payload to the vicinity of those tumor cells. As indicated herein, the Nano-ADCs of this disclosure can have increased tumor accumulation, such as shown in FIG. 21 , that can enhance tumor targeted cytotoxicity while can have potential to reduce non-specific cytotoxicity toward non-tumor cells therefore reducing side effects.Such increased tumor specific tissue accumulation can be a part of the nanoparticle- mediated cytotoxicity to tumor cells mentioned herein.

[0286] An additional advantage of the pharmaceutical composition of this disclosure is that multiple molecules of a drug (i.e. , PBA) can be linked to the TBA. The polymer can be a branched polymer and can be linked to the TBA, such as a monoclonal antibody, at only a limited number of sites, such as at one cysteine or one lysine residue site, while at the same time linked to multiple molecules of the drug (see at least FIG. 3A-FIG. 3D, FIG. 4A-FIG. 4D, FIG. 5, FIG. 6A). This can significantly improve the PBA to TBA ratio, such as drug to antibody ratio increasing payload of the ADC. In addition, the drugs (PBAs) can be positioned far away from the TBA, such as an antibody, so the drugs can be more readily available due to less stereo hindrance.

[0287] Yet another advantage of the pharmaceutical composition of this disclosure is that the linkers L1 and L2 can be the same or different. By selecting different linkers L1 and L2, TBA and PBA can be selectively dissociated and selectively deliver the PBA to target cells.

[0288] Yet another advantage of the pharmaceutical composition of this disclosure is that the polymer can provide an environment to enhance water solubility of the water poorly soluble APIs that can potentially increasing bioavailability or effect in treating diseases.

[0289] Yet another advantage of this disclosure is that the Nano-ADCs can have similar cytotoxicity towards tumor cells having high levels of tumor antigen (AgHightumor cells), while the Nano-ADCs can provide better AgLowtumor cytotoxicity towards tumor cells having low, very low or free from the tumor antigen (collectively, AgLowtumor cells) (FIG. 17A and FIG. 17B).

[0290] In some cases, this disclosure is also directed to a nanoaggregate comprising a therapeutic bioactive agent and a polymer, wherein the nanoaggregate have particle sizes in a range of from 1 nm to 140 nm, and the nanoaggregate can comprise a tumor cytotoxicity to tumor cells and a non-tumor cytotoxicity to non-tumor cells, wherein the tumor cytotoxicity comprises AgHightumor cytotoxicity to tumor cells having the tumor antigen (AgHightumor cells) and AgLowtumor cytotoxicity to tumor cells having low, very low or free from the tumor antigen (AgLowtumor cells), and the pharmaceutical composition comprises the AgLowtumor cytotoxicity to the AgLowtumor cells in the absence of the AgHightumor cells.

[0291] In some cases, in a range of from 20% to 100% of the nanoaggregates have particle sizes in a range of from 2 nm to 100 nm.

[0292] In some cases, in a range of from 40% to 100% of the nanoaggregates have particle sizes in a range of from 2 nm to 80 nm.

[0293] In some cases, in a range of from 40% to 100% of the nanoaggregates have particle sizes in a range of from 2 nm to 80 nm, 2 nm to 60 nm, 2 nm to 40 nm, 2 nm to 30 nm, 2 nm to 20 nm, 2 nm to 25 nm, 2 nm to 20 nm, 2 nm to 8 nm, and 2 nm to 4 nm.

[0294] In some cases, this disclosure is further directed to a use of a bioactive composition, Nano-ADC or a pharmaceutical composition disclosed herein for manufacturing of a medicament for treating or preventing a disease, wherein the bioactive composition, Nano-ADC or the pharmaceutical composition can comprise: a PBA, a TBA comprising an antibody or a part thereof having binding affinity to a target molecule that comprises a tumor antigen (Ag); a polymer, a linker, optionally a coupler, or a combination thereof, wherein the PBA is covalently linked to the polymer, the linker, or optionally, the coupler when present, or a combination thereof.

[0295] The PBA, TBA, polymer, linker, and coupler disclosed herein can be suitable.

[0296] In some cases, the PBA can comprise a formula selected from Formula ID 1 , 1-1 , 1-2, 2, 2-1 , 2-2, 3, 3-1 , 3-2, and 1001-1135, a pharmaceutically acceptable salt thereof, a solvate thereof, a prodrug thereof, an isomer thereof, an isotope derivative thereof, and a combination thereof.

[0297] ln some cases, the PBA can comprise Formula ID 1 , 1-1 , 1-2, 2, 2-1 , 2-2, 3, 3-1 , 3-2, and 1001-1135, one or more additional cytotoxic agents disclosed herein, a pharmaceutically acceptable salt thereof, a solvate thereof, a prodrug thereof, an isomer thereof, an isotope derivative thereof, and a combination thereof.

[0298] In some cases, the Nano-ADC comprises the coupler that can comprise Formula ID 201 , 202, 203, 204, 205, 206, 207, 208, 209, or a combination thereof.

[0299] In some cases, the linker can comprise a cleavable linker and can comprise a Linker L1 , a Linker L2, or a combination thereof, wherein the Linker L1 is covalently linked to the TBA and covalently linked to a reacted first terminal group of the polymer, the Linker L2 is covalently linked to the PBA and covalently linked to a reacted second terminal group of the polymer. In some cases, the linker can comprise a cleavable linker selected from Formula ID 101 , 102, 103, 104, 105, 106, 107, 108, 109, 110, 111 , 112, 113, 114, 115, 116, 117, 118, 119, 120, 121 , 122, 123, 124, 125, 126, 127, 128, 129, 130, 131 , 132, 133, 134, 135, 136, 137, 138, 139, 140, 141 , and a combination thereof.

[0300] In some cases, the TBA can comprise an antibody disclosed herein.

[0301] In some cases, the polymer can comprise a peptide, a polyethylene glycol (PEG), polyoxazoline (POX), or a combination thereof, wherein the polyoxazoline (POX)comprises a linear portion, a branched portion, or a combination thereof, and wherein the polyoxazoline (POX) comprises poly(2-methyloxazoline), poly(2-ethyloxazoline), poly(2- propyloxazoline), poly(2-isopropyloxazoline), poly(2-substituted oxazoline), or a combination thereof, as disclosed herein.

[0302] In some cases, polymer can comprise a single amino acid residue, dipeptide, tripeptide, or polypeptide. In some cases, as mentioned above, the polymer can comprise a peptide comprising in a range of from 1 to 100 amino acid residues. In some cases, a polymer can comprise a lysine, glutamic acid, glutamine, arginine, valine, or a combination thereof.

[0303] In some cases, suitable to the use disclosed herein, the Nano-ADC can comprise a formula selected from Formula ID 3001-3074, a pharmaceutically acceptable salt thereof, a solvate thereof, a prodrug thereof, an isomer thereof, an isotope derivative thereof, and a combination thereof;

[0304] Suitable to the use disclosed herein, the disease is cancer.

[0305] The instant disclosure now will be exemplified in the following non-limiting examples.EXAMPLES

[0306] The present disclosure is further defined in the following Examples. It should be understood that these Examples, while indicating preferred embodiments of the disclosure, are given by way of illustration only. From the above discussion and these Examples, one skilled in the art can ascertain the essential characteristics of this disclosure, and without departing from the spirit and scope thereof, can make various changes and modifications of the disclosure to adapt it to various uses and conditions.MATERIALS AND MEASUREMENTSNanoparticle Measurement

[0307] The size of various polymers, nanoaggregates, as well as antibody-polymer-drug nanoaggregates were measured by a dynamic light scattering (DLS) method using a Malvern Zetasizer Nano-ZS Zen3600 particle size analyzer (Malvern Panalytical Inc., Westborough, MA 01581 , USA).Drug-antibody ratio (DAR)

[0308] Drug-antibody ratio (DAR) of an ADC can be calculated from the amounts of the antibody and the drug (PBA). In one method, the amount of antibody and the PBA can be in nanomole (nmole):DAR=PBA (nmole) / Ab (nmole).

[0309] Drug-antibody ratio (DAR) of an ADC can also be calculated from measurement data with the formula (herein “measured DAR”):DAR=8x((A379 / A280) / 0.46) wherein, DAR is Drug-antibody ratio, A379 is the absorbance at 379 nm, A280 is the absorbance at 280 nm.

[0310] Drug-antibody ratio (DAR) of an ADC can also be referred to as a nominal DAR based known coupling sites on an antibody and the coupling method. For example, for an antibody having 4 cysteine residues, a nominal DAR can be 1-4 and for an antibody having 4-pairs of cysteine residues (4 pairs of S-S bonds) that can couple up to 8 drugs, a nominal DAR can be 1-8.ADC Preparation:

[0311] A representative reaction scheme is briefly described here: 21 nmol of an antibody was reacted with 210 nmol of TCEP (tris(2-carboxyethyl)phosphine) in PBS, pH 6.0 with 5 mM of EDTA, reacted at 37 °C for 1 hour, followed by the addition of 420 nmol of a linker payload molecule having one of the Formula ID 2001-2040 prepared herein, or a combination thereof, incubated at room temperature for 1 hour. The reaction was terminated with 840 nmol of N-Acetyl-L-cysteine at room temperature for 20 minutes. The mixture was purified with a gel filtration chromatography column and filtration. The conjugate collected was at about 50kD molecular weight (MW) cut-off and was characterized with HIC (Hydrophobic Interaction Chromatography), SEC (Size-exclusion chromatography) and UV. Alternatively, a coupler can be first linked to an antibody via reductive coupling and then linked to a PBA, directly or via a polymer, a linker or a combination thereof. The conjugated products can be purified as described above.Procedure for producing PEOX polymer having VARIOUS EOX UNITS:

[0312] PEOX polymer having different EOX units (such as PEOX2, PEOX4, PEOX6, PEOX8, PEOX10, PEOX12, PEOX16, PEOX18, PEOX20, PEOX20, and PEOX24 (y=2- 24)) and different MW can be prepared by different initiator to monomer ratios. In brief, 2- ethyl-2-oxazoline, methyl-bromo-acetate, and acetonitrile under the nitrogen protection were reacted inside a heat block set at 92 °C on a hot plate with magnetic stirring and reacted for overnight. After polymerization, the solution was cooled to room temperature,and the resulting polymer was then precipitated out from MTBE. The wet polymer was vacuum dried at room temperature over night to produce a crude polymer product. The crude polymer product was purified, dialyzed, dried by rotovap to produce the final HO2C- CH2-(PEOX)yOH PEOX polymer. In a PEOX preparation having a ratio of initiator to monomer = 1 :18, the structure was confirmed by the1H-NMR and n was determined to be 18 EOX units (y=18). Polydispersity of the polymer was determined by SEC. In some polymer preparations, polydispersity indices were about 1.07.EXAMPLE 1

[0313] Modified exatecan Formula ID 1 -1 can be synthesized using the general procedure above and schematically shown in following reactions.wherein, v=1 -100; w=1-1000;R= H, D, C1 -22 alkyl group, aromatic group, or substituted aromatic group, C1-C100 saturated or unsaturated hydrocarbon (including cyclic hydrocarbon as defined herein), C1-C100 saturated or unsaturated hydrocarbon having at least one D, C1-C100 saturated or unsaturated hydrocarbon having at least one heteroatom, C1-C100 saturated or unsaturated hydrocarbon having at least one heteroatom and at least one D.

[0314] Formula ID 1006 (EE) was produced as described in detail below:

[0315] Step 1 a &1 b: To an ice-cooled solution of 2-((tert- butyldimethylsilyl)oxy)ethanamine (8.5 grams, 48 mmol) in dichloromethane (90 ml_), triethylamine (9 mL) were added, followed by a solution of benzyl bromoacetate (5 g, 21 mmol) over 5 min. After stirring in the ice bath for 30 min, triethylamine (5 mL) was added, followed by propionic anhydride (7.5 g, 57 mmol) in three portions over 3 min. The resulting mixture was stirred at room temperature (RT) for 10 min, then water (50 mL) was added. The organic layer was separated and concentrated via rotary evaporation. The crude product was purified by silica gel chromatography, eluted with mixed heptaneand ethyl acetate to produce 2.3 g of the product as an oil with MS molecular mass plus proton ion (ESI [M+H]) was 380.

[0316] Step 2: To a solution containing 230 mg of the product from step 1 b in EtOH (15 mL), 5% Pd / C (150 mg) was added. The solution was hydrogenated under 100 psi of H2 at RT for 3 hrs, and the mixture was filtered through a celite pad, concentrated on rotavapor to 160 mg of desired product, which was used for next step without further purification.

[0317] Step 3: To an ice-cooled mixture of EDC-HCI (53 mg) in DMF (1 mL), Oxyma Pure (40 mg) was added, followed by a solution of the product from step 2 (80 mg) in dichloromethane (1 mL). The resulting mixture was stirred in the ice bath for 5 min to form a light yellow solution. In a separate vial, exatecan mesylate (100 mg) and DMF (2 mL) were mixed and the mixture was cooled in an ice bath, then DI PEA (0.4 mL) was added, followed by addition of the light yellow solution prepared before. The mixture was stirred at RT overnight. The resulting solution was diluted with dichloromethane (20 mL), washed with 20% LiCI aqueous solution (10 mL), followed by saturated aqueous solution of NH4CI. The organic phase was concentrated on rotavapor to produce a crude liquid product, which was used for the next step without further purification.

[0318] Step 4: To the crude liquid product from step 3, water (5 mL), THF (5 mL) and acetic acid (15 mL) were added. The mixture was stirred at RT for 5 hrs, and then the solution was concentrated on rotavapor to a produce a product with a volume of about 2 mL. The product was loaded on a 40 g of C18 column, eluted with 10% CH3CN in water for 5 min, then increased to 70% over 10 min to produce about 57 mg of desired product as grey solid, ESI, [M+H]+, 593.56.

[0319] The reaction scheme is outlined here:(Formula ID 1006), a modified exatecan having an ethyl group (“EExa” or “EE”).

[0320] Following modified exatecans were synthesized using the above-described reactions: Formula ID 1001-1016, and 1019-1029.

[0321] Modified exatecans having Formula ID 1030-1045, 1048-1058,1059-1074, 1077-1087, 1088-1103, 1106-1116, and 1117-1118, can be synthesized using similar reactions.EXAMPLE 2

[0322] The modified exatecan Formula ID 2-1 can be synthesized using following reactions:R’ each is independently H, D, C1-22 alkyl group, aromatic group, substituted aromatic group, C1-C100 saturated or unsaturated hydrocarbon (including cyclic hydrocarbon as defined herein), C1-C100 saturated or unsaturated hydrocarbon having at least one D, C1-C100 saturated or unsaturated hydrocarbon having at least one heteroatom, C1-C100 saturated or unsaturated hydrocarbon having at least one heteroatom and at least one D, or a combination thereof; and X is O, S, or NH.

[0323] A modified exatecan Formula ID 2-2 was synthesized using the same reactions:

[0324] Additional modified exatecan Formula ID 1127-1131 can be synthesized using the same reactions.EXAMPLE 3

[0325] The modified exatecan Formula ID 3 can be synthesized using following reactions:ExatecanR” is independently H, D, C1 -22 alkyl group, aromatic group, substituted aromatic group, C1 -C100 saturated or unsaturated hydrocarbon (including cyclic hydrocarbon as defined herein), C1-C100 saturated or unsaturated hydrocarbon having at least one D, C1-C100 saturated or unsaturated hydrocarbon having at least one heteroatom, C1 -C100 saturated or unsaturated hydrocarbon having at least one heteroatom and at least one D, or a combination thereof; and X is O, S, or NH.

[0326] A modified exatecan Formula ID 3-2 was synthesized using the same reactions:

[0327] Additional modified exatecan Formula ID 1132-1135 can be synthesized using the same reactions.EXAMPLE 4

[0328] A Nano-ADC comprising a Mal-PEG coupler having a 24 unit-PEG polymer, GGFG cleavable link and a modified exatecan having an ethyl group (EE or EExa) (Formula ID 2010) was synthesized according to the reaction scheme below:(Formula ID 2010).

[0329] The product above with Formula ID 2010 was then reacted with a monoclonal antibody (mAb) HER2 receptor antagonist trastuzumab biosimilar (KANJINTI®) (herein “Kan” or “Kan antibody”) to form a Nano-ADC having Formula ID 3024K (also referred to as Kan-PEG24-GGFG-EE, or Kan-Mal-PEG24-GGFG-EE) with n in a range of from 5-10 to 1 :(general Formula ID 3024).CT

[0330] The produced Nano-ADC Formula ID 3024K with the Kan antibody that had a nominal DAR of 8 (FIG. 14A-FIG. 14B).EXAMPLE 5

[0331] An ethyl modified exatecan (EE or EExa) with a cleavable linker GGFG, herein referred to as “PEGm-GGFG-EE” was synthesized using the following reactions.CTO 6 Exa(Formula ID 2031).

[0332] The term “Exa” shown in the reactions above refers to an un-modified exatecan.

[0333] The Formula ID 2031 was then reacted with an anti-HER2 monoclonal antibody via a cystine-based conjugation to form an ADC.EXAMPLE 6

[0334] Using the similar reactions, follow modified exatecans (mExa) with aVAPAB linker can be synthesized: Formula ID 2032 from mExa Formula ID1-2, Formula ID 2033 from mExa Formula ID 3-2 and Formula ID 2034 from mExa Formula ID 2-2, respectively, wherein m is an integer in a range of from1 - 100,(Formula ID 2033),CT(Formula ID 2035).

[0335] The Formulas 2032, 2033 and 2034 can each be reacted with an anti- HER2 monoclonal antibody trastuzumab biosimilar (KANJINTI®) (Kan), to form a corresponding ADC comprising the mExa having Formula ID 3021 , 3022, and 3023, respectively.EXAMPLE 7

[0336] Using the modified exatecan and the reactions described above, following Nano-ADCs were synthesized with m=24 and n=4-10, i.e., DAR (Drug-to-Antibody Ratio) in a range of from 4-10 to 1 : Formula ID 3004, 3005, 3006, 3007, 3008, 3009, 3010 (FIG. 10 D-FIG. 10J), respectively.EXAMPLE 8

[0337] Modified exatecan Formula was prepared according to reaction scheme described above. A Nano-ADC was synthesized with a SMCC coupler, GGFG cleavable link, the mExa (EE), and the monoclonal antibody (mAb) HER2 receptor antagonist trastuzumab biosimilar (KANJINTI®) (Kan), without any PEG polymer, according to Formula ID 301 1 (FIG. 10K), n=4-10, i.e., DAR in a range of from 4-10 to 1 : Kan-SMCC-GGFG-EE:CT(Formula ID 3025) (FIG. 12H).

[0338] Similar Nano-ADC was prepared using trastuzumab biosimilar (KANJINTI®) (Kan), 24 unit-PEG polymer (PEG24), a VAPAB cleavable link (VAPAB) and a modified exatecan (mExa or EE) to form a Nano-ADC herein referred to as “Kan-PEG24-VAPAB-EE” (Formula ID 3026) (FIG. 121).EXAMPLE 9

[0339] A bioactive composition comprising a MC coupler, a tripeptide Lys-Val- Ala polymer, a PEOX polymer side chain with an un-modified exatecan (E) as a PBA, without the para-aminobenzyl group, was synthesized using the reactions described herein:CTCTFormula ID 2036, and y = 1-24 (also referred to as MC-Lysine-PEOX-VA-E).

[0340] A Nano-ADC Formula ID 3038 was produced by conjugating a Kan antibody with the Formula ID 2036 with y=18, n=8 (FIG. 12U).EXAMPLE 10

[0341] A bioactive composition comprising a MC coupler, an amino acid glutamic acid, a GGFG linker, a PEOX polymer side chain with a modified exatecan (EE) and MMAF as dual payload PBAs, was synthesized using the reactions described herein:CTFormula ID 2029, y=1-24.EXAMPLE 11

[0342] A bioactive composition comprising a MC coupler, lysine, a GGFG linker, multiple polymer side chains comprising PEOX polymers with an un-modified exatecan (E) and MMAF as dual payload PBAs, was synthesized using the reactions described herein (Formula ID 2030):CTTHF, 0°C step 3 step 2 step 4CTFormula ID 2030, y and y’= 1 -24, y and y’ can be the same or different. It is also referred to as L(PEOX)-PE-VL(PEOX)-p-MMAE.EXAMPLE 12

[0343] A bioactive composition comprising a SMCC coupler, a GGFG linker, a peptide comprising Val-Ala, a polymer side chain comprising PEOX polymer with an un-modified exatecan (E) as a payload PBA, was synthesized using the reactions described herein (Formula ID 2021 ):CTFormula ID 2021 , y is 1-24. It is also referred to as SMCC-L(PEOX)-VA-E.EXAMPLE 13CT

[0344] A bioactive composition comprising a SMCC coupler, a polymer comprising amino acid lysine, a polymer side chain comprising PEOX polymer with MMAF as a payload PBA, was synthesized using the reactions described herein (Formula ID 2024):CTFormula ID 2024, y=1 -100. It is also referred to as SMCC-L(PEOX)-p-MMAF.

[0345] Various products were prepared with y=4, 8, 10, 12, 18, and 24.EXAMPLE 14

[0346] A bioactive composition comprising a Mal-coupler, a polymer comprising PEG, and a modified exatecan having Formula ID 1005 (TFE) was synthesized using the reactions described herein to produce Formula ID 2038):CTCT(Formula ID 2038, Mal-PEG-GGFG-TFE), m = 2-100.

[0347] Various preparations were produced with m = 2, 4, 8, 10, 12, 14, 16, 18, 20 and 24, i.e., Mal-PEG2-GGFG-TFE, Mal-PEG4-GGFG-TFE, Mal-PEG8- GGFG-TFE, Mal-PEG10-GGFG-TFE, Mal-PEG12-GGFG-TFE, Mal-PEG14- GGFG-TFE, Mal-PEG16-GGFG-TFE, Mal-PEG18-GGFG-TFE, and Mal-PEG24-GGFG-TFE.

[0348] Formula ID 2038 was conjugated to the Kan antibody to produce NanoADC formula ID 3029 (FIG. 12L).EXAMPLE 15-20

[0349] Synthesis of Formula ID 2005, 2025, 2026, 2027, 2039, and 2040.EXAMPLE 15

[0350] Formula ID 2005 was produced via reactions described below:CTCTMALdPEG24NHS step 5(Formula ID 2005, y=10). Various products were prepared with y=1 , 2, 4, 8, 10, 12, 18, and 24 using the same procedure described herein.EXAMPLE 16

[0351] Formula ID 2025 was produced via reactions described below:CT(Formula ID 2025, y was 18 on average). Various products were prepared with y=1 , 2, 4, 8, 10, 12, 18, and 24 using the same procedure described herein.EXAMPLE 17CT

[0352] Formula ID 2026 was produced via reactions described below:CT(Formula ID 2026, MC-VAPAB-o-PEOX-MMAE, y average 18). Various products were prepared with y=1 , 2, 4, 8, 10, 12, 18, and 24 using the same procedure described herein.EXAMPLE 18Pd / C, H2, MeOH step 2CTCT(Formula ID 2027, y and y’ =18). Various products were prepared with y and y’ =1 , 2, 4, 8, 10, 12, 18, and 24 using the same procedure described herein with y and y’ can be the same or different.EXAMPLE 19

[0354] Formula ID 2039 was produced via reactions described below:CTCTDBU (0.8 eq)DMFHOBt-H2OEDC-HCI step 3CT(Formula ID 2039), y=10. Various products were prepared with y=1 , 2, 4, 8, 10, 12, 18, and 24, using the same procedure described herein.EXAMPLE 20

[0355] Formula ID 2040 was produced via reactions described below:step 3CT(Formula ID 2040), y=1-100. It is also referred to as MC-L(PEOX)-p-MMAF.Various products were prepared with y=1 , 2, 4, 8, 10, 12, 18, and 24.EXAMPLE 21

[0356] Nano-ADCs having Formula ID Kan-2001 through Kan-2038 were prepared using a mAb trastuzumab (HER2 receptor antagonist monoclonalCT antibody biosimilar, KANJINTI®, under respective trademark) (herein “Kan”) conjugated with Formula ID 2001 through 2040).

[0357] Particular Nano-ADCs also included Formula ID 3025 (Kan-MC-GGFG- Formula ID 1006), 3027 (Kan-SMCC-L(PEOXI O)-p-E), 3028 (Kan-Mal-PEG24- GGFG-Formula ID 1006), 3029 (Kan-Mal-PEG24-GGFG-Formula ID 1005, TFE), and 3030 (Kan-SMCCC-L(PEOX20)-p-E) (Table 1).

[0358] Nano-ADCs having Formula ID IF-2001 through IF-2037 were prepared using ifinatamab (anti-human B7-H3 (CD276) monoclonal antibody) (herein “I Fl”, or “B7H3”) conjugated with Formula ID 2001 through 2040.

[0359] Particular Nano-ADCs also included Formula ID 3031 (B7H3-SMCC- L(PEOX18)-p-E), 3032 (IFI-SMCC-L(PEOXI O)-p-E), 3033 (Ab[-(SMCC- L(PEOX10)-p-E)n, -(Mal-PEG2-VC-MMAE)n’], a B7H3 antibody ifinatamab conjugated to two biomolecules -SMCC-L(PEOX10)-p-E and -PEG24-VC- MMAE) (FIG. 12P), 3034 (Ab[-(SMCC-L(PEOX)-p-E)n, -(Mal-PEG2-VC- MMAE)n’], an antibody conjugated with two biomolecules -SMCC-L(PEOX)-p- E and -Mal-PEG2-VC-MMAE, with y=4, 6, 8, 10, 12, 18, 24, and n’:n ratio in a range of from 1 :8 to 8:1 ) (FIG. 12Q) and 3035 (a B7H3 antibody ifinatamab conjugated to two biomolecules -SMCC-L(PEOX10)-p-E and -PEG24-GGFG- TFE (FIG. 12R). Nano-ADCs Formula ID 3033 and 3035 are also referred to as Nano-ADC having mixed PBAs. Formula ID 3033 products had various n’:n ratios in a range of from n’:n=4:1 to 1 :4. Formula ID 3035 products had various n’:n ratios in a range of from n’:n=8:1 to 1 :8.

[0360] Nano-ADC Formula ID 3036 Kan[-(SMCC-L(PEOX)-p-E)n, -(Mal-PEG2- VCP-MMAE)n’] was produced by conjugating trastuzumab biosimilar (KANJINTI®, Kan) antibody with mixed biomolecules -SMCC-L(PEOX10)-p-E and -Mal-PEG2-VC-MMAE (FIG. 12S), with y=4, 6, 8, 10, 12, 18, 24, and n’:n ratio of 3:1. Formula ID 3036 products had various n’:n ratios in a range of from n’:n=8:1 to 1 :8.

[0361] Nano-ADC Formula ID 3037 Kan-(SMCC-L(PEOX)-VP-E)n (FIG. 12T) was produced by conjugating trastuzumab biosimilar (KANJINTI®, Kan) antibody with a biomolecule Formula ID 2003 (-SMCC-L(PEOX)-VP-E), with y=10, n=8.

[0362] Nano-ADC Formula ID 3038 Kan-(SMCC-L(PEOX)-VA-E)n (FIG. 12U) was produced by conjugating the trastuzumab biosimilar (KANJINTI®, Kan)CT antibody with a biomolecule Formula ID 2036 (-SMCC-L(PEOX)-VA-E) with y=18, n=8.EXAMPLE 22

[0363] Cytotoxicity of payload (PBA) without antibody was measured in vitro with cell viability assay using LNCaP cell line that is a prostate cancer cell line having low HER2 expression levels. Data are shown in FIG. 15: un-modified Exatecan (E) (solid circle), Dxd (deruxtecan) (solid square), TFE (Formula ID 1005) (solid diamond), PrE (Formula I D 1004) (overlaid square and solid circle), BzE (Formula ID 1012) (open triangle), cHME (Formula ID 1013) (small solid dot), nBE (Formula ID 1014) (+), iBE (Formula ID 1015) (x), nPE (Formula ID 1016) (open circle), bPE (Formula ID 1011) (overlaid open square and asterisk) (FIG. 15). Modified exatecans (TFE, PrE, BzE, cHME, nBE, iBE, nPE and bPE) had similar cytotoxicity in vitro in the low HER2 cells compared to the unmodified exatecan and the commercially available modified exatecan deruxtecan (Dxd) in the drug concentrations tested.EXAMPLE 23

[0364] Cytotoxicity of the Nano-ADCs prepared above were measured using conventional in vitro cell-based assay with SKBR-3 cell line that is a human breast cancer cell line overexpressing the HER2 (Neu / ErbB-2) (the human epidermal growth factor receptor 2) gene product HER2 and NCI-N87 cells that are gastric cancer cells with high expression of the human epidermal growth factor receptor 2 (HER2).

[0365] Nano-ADC produced in Example 4 (Kan-PEG24-GGFG-EE, Formula ID 3024K, with monoclonal antibody trastuzumab biosimilar (KANJINTI®), shown as Kan) was tested. HER2 receptor antagonist trastuzumab biosimilar (KANJINTI®, under respective trademark) without any linked payload was used as a negative control and ADC ENHERTU® (trastuzumab deruxtecan (Dxd), Daiichi Sankyo / AstraZeneca under respective trademark), was used as a comparative.

[0366] The cells were subject to increasing drug concentrations of the drug tested and cell viability was measured using traditional methods. Data are shown in FIG. 16A for SKBR-3 cells and FIG. 16B for NCI-N87 cells.CT

[0367] The Nano-ADC 3024K (Formula ID 3024K) (open circle) showed cytotoxicity similar to that of ENHERTU (solid square) that exhibited cytotoxicity to the cells tested when compared to the control antibody KANJINTI alone (solid circle).EXAMPLE 24

[0368] The cytotoxicity of the Nano-ADCs prepared above were measured using mouse models xenografted with the HCT116 tumor cells derived from colon cancer and HT29 cells derived from colorectal adenocarcinoma that are considered to have low HER2 expression levels.

[0369] About 2-3 million tumor cells were cultured and harvested to inoculate into each SCID mice subcutaneously. Tumors from the tumor cells were allowed to grow to 150-200 mm3(around 10 days).

[0370] Following ADC samples were administered to the mice through tail vein on Day 0 once the tumors were grown to about 150-200 mm3in size: Nano- ADC Formula ID 3027 (Kan-SMCC-L(PEOXI O)-p-E), 3029 (Kan-Mal-PEG24- GGFG-TFE), and controls were injected to the HCT 116 mice at 3 mg / kg. A 5% glucose solution was used as a negative control (shown as “Control”). Body weight and tumor size were recorded twice a week starting from Day 0. Relative tumor sizes were normalized to day 0 and plotted against the treatment days. Data from Nano-ADCs comprising modified exatecans were compared to commercially available trastuzumab deruxtecan (ENHERTU®) (shown as “Enhertu”) (FIG. 17A).

[0371] The commercially available ENHERTU exhibited no significant inhibition to the tumor growth compared to Control. The Nano-ADCs tested showed significantly enhanced inhibition of tumor growth in these mouse models when compared to that of the ENHERTU’s (p<0.03): Nano-ADCs of Formula ID 3027 and 3029 exhibited greater inhibition in tumor growth with tumor sizes of about 1 / 3 (Formula ID 3027) and 1 / 2 (Formula ID 3029) compared to the ENHERTU at day 24 (FIG. 17A).

[0372] Following ADC samples were administered to HT29 mice through tail vein on Day 0 at 3 mg / kg: Nano-ADC Formula ID 3036 (Kan[-(SMCC- L(PEOX10)-p-E)n, -(Mal-PEG2-VC-MMAE)n’], with the antibody being trastuzumab biosimilar (KANJINTI®, a Nano-ADC having mixed PBAs of un-CT modified exatecan and MMAE, y=10). Commercially available ENHERTU and Formula ID 3080 (HlgG[-(SMCC-L(PEOX10-p-E)n, -(Mal-PEG2-VC-MMAE)n’], an ADC similar to Formula I D 3036 having mixed PBAs of un-modified exatecan and MMAE, and non-specific human IgG as an antibody), y=10, n=1 and n’=3, were used as comparatives. Nano-ADC of Formula ID 3036 exhibited greater inhibition in tumor growth with minimum tumor growth on Day 9 compared to the ENHERTU (p=0.03) and the ADC with non-specific human IgG (Formula ID 3080) (FIG. 17B).EXAMPLE 25

[0373] The cytotoxicity of the Nano-ADCs prepared above were measured using mouse models xenografted with NCI-N87 that is a gastric cancer cell line, having high HER2 expression levels.

[0374] The tumor cells were inoculated into SCID mice as described above. Nano-ADCs, Formula ID 3027 (Kan-SMCC-L(PEOXW)-p-E), 3024K (Kan- PEG24-GGFG-EE), both with the trastuzumab biosimilar (KANJINTI®, “Kan”) antibody, Control (5% glucose solution) and Enhertu (trastuzumab deruxtecan, ENHERTU®) were individually injected to the mice as described above at 1 mg / kg. Tumor sizes were measured on the days indicated as described above. Data are shown in FIG. 18A. Control (solid circle), Enhertu (solid square), Formula ID 3027 (solid diamond), and Formula ID 3024K (open circle). The Nano-ADCs and Enhertu all had significant inhibition of tumor growth compared to Control. Nano-ADCs Formula ID 3024K having the PEOX polymer exhibited greater inhibition in tumor growth with negligible tumor growth through the 30- day measurement time period (FIG. 18A).

[0375] In a separate measurement, cytotoxicity of Nano-ADC Formula ID 3035 (Kan[-(SMCC-L(PEOX10)-p-E)n, -(Mal-PEG24-GGFG-TFE)n’] having mixed payload PBAs un-modified exatecan and modified exatecan TFE, with the trastuzumab biosimilar (KANJINTI®, “Kan”) antibody was determined using the same NCI-N87 cells and SCID mice as described above. Data are shown in FIG. 18B (Formula ID 3035, open circle). Nano-ADCs Formula ID 3035 exhibited greater inhibition in tumor growth with negligible tumor growth through the 30-day measurement time period (FIG. 18B) that was significantly different from Enhertu (p=0.005).CTEXAMPLE 26

[0376] The cytotoxicity of the Nano-ADCs prepared above were measured using mouse models xenografted with NCI-H446 tumor cells that is a small cell lung cancer cell line having medium B7H3 expression levels, and NCI-H526 tumor cells that is a small cell lung cancer cell line having high B7H3 expression levels.

[0377] The tumor cells were cultured, harvested and inoculated into the mice as described above. Following ADC samples were administered to the mice through tail vein on Day 0 once the tumors were grown to about 150-200 mm3in size: Nano-ADC Formula ID 3033 (IFI-[-(SMCC-L(PEOX10)-p-E)n, -(Mal- PEG2-VC-MMAE)n’], with y=10 and Ab= ifinatamab (anti-human B7-H3 (CD276) monoclonal antibody, herein “I Fl”), and l-Dxd (Ifinatamab deruxtecan, a comparative ADC) were injected to the NCI-H446 mice (FIG. 19A) and NCI- H526 mice (FIG. 19B), respectively, at 1 mg / kg. A 5% glucose solution was used as a negative control (shown as “Control”). Body weight and tumor size were recorded twice a week starting from Day 0. Relative tumor sizes were normalized to day 0 and plotted against the treatment days. Data are shown in FIG. 19A and FIG. 19B. The Nano-ADC Formula ID 3033 conjugated with ifinatamab had significantly greater inhibition of tumor growth with negligible tumor growth through the 30-day measurement time period compared to l-Dxd (p=0.03) and Control.EXAMPLE 27

[0378] The cytotoxicity of the Nano-ADCs prepared above were measured via multiple injections using SCID mouse models xenografted with the HCT116 tumor cells derived from colon cancer that are considered to have low HER2 expression levels (HER2Low) (FIG. 20).

[0379] The HT116 tumor cells were inoculated to SCID mice as described above. After allowing the tumors to grow for about 10 days, traditional ADC ENHERTU and Nano-ADC Formula ID 3027 with the Kan antibody, PEOX polymer side chain and un-modified exatecan (Kan-SMCC-LPEOX10-p-E) (DAR=8) and Formula ID 3037 with the Kan antibody, PEOX polymer side chain and un-modified exatecan (Kan(-SMCC-LPEOXI O-VP-E)n) (DAR=8) wereCT injected to the mice at the indicated time points, i.e., day 1 , day 15 and day 29 (indicated at the time axis), each at 3 mg / kg.

[0380] Representative data are shown in FIG. 20: (1 ) Control (solid circle) with injection of 5% glucose solution; (2) two ENHERTU injections on day 1 and day 15, and 1 injection of Formula ID 3027 on day 29 (open circle); (3) one ENHERTU injection on day 1 and two injections of Formula ID 3027 on day 15 and day 29 (solid square); (4) one ENHERTU injection on day 1 and two injections of Formula ID 3037 on day 15 and day 29 (open square); (5) three injections of Formula ID 3027 on day 1 , day 15 and day 29 (open triangle); and (6) three injections of Formula ID 3037 on day 1 , day 15 and day 29 (solid triangle). The mice with control were terminated earlier due to tumor growth.

[0381] Data indicated that the Nano-ADCs Formula ID 3027 (open triangle) and 3037 (solid triangle) had significant inhibition of tumor growth compared to Control (solid circle), compared to two ENHERTU injections (open circle) (p<0.01 ).

[0382] The tumor sizes of injections with two times Nano-ADC plus one time ENHERTU (solid square) were also significantly reduced compared to one time Nand-ADC injection and two time ENHERTU injections (open circle) (p=0.013).

[0383] Data showed that the Nano-ADC exhibited a tumor inhibition activity to inhibit tumor growth after failed treatment with an ADC fam-trastuzumab deruxtecan-nxki (ENHERTU®) indicating potential effect in inhibiting tumor growth in HER2Lowtumors or ENHERTU resistant tumors, especially when the Nano-ADC was injected in an early stage of tumor growth.EXAMPLE 28

[0384] Tissue distribution of the PBA measured according to the procedure described here.

[0385] About 10 mg / kg of different Nano ADC and a comparative ADC Enhertu were administrated into NCI-N87 tumor baring SCID mice through tail vein as described above. About 24 hours after the administration, whole blood was harvested through inferior vena cava of the mice and the plasma was separated to produce plasma samples (Plasma). Tumors were harvested and processed to produce tumor samples (Tumor). The plasma and tumor samples were measured for exatecan or deruxtecan using Liquid chromatography-massCT spectrometry (LC / MS) with un-modified exatecan or deruxtecan as standard. Data are shown in FIG. 21. In plasma samples, both mice injected with Enhertu (Enhertu)) and Nano-ADC Formula ID 3027 (Kan-SMCC-L(PEOXW)-p-E having the PEOX polymer and the Kan antibody) (Formula ID 3027) showed very low amounts of exatecan (“plasma” in FIG. 21) and had no significant difference between the Enhertu injected and the Formula ID 3027 injected mice (p=0.3). In tumor samples, the mice injected with Nano-ADC Formula ID 3027 had significantly high amount of exatecan (p=0.02) compared to that of the Enhertu injected mice (“Tumor” in FIG. 21) indicating significantly increased accumulation of the payload in the tumor tissue.COMPARATIVE EXAMPLES

[0386] The original exatecan was also used to produce an ADC that has conjugated same antibody trastuzumab biosimilar (KANJINTI®) (Kan), coupler, cleavable linker GGFG, and PEG24, herein referred to as “Kan-PEG24-GGFG- Exa” or “Kan-PEG24-GGFG-E” as a comparative.

[0387] Additional comparative example was prepared to have conjugated same antibody trastuzumab biosimilar (KANJINTI®) (Kan), coupler, cleavable linker VAPAB, and PEG24, herein referred to as “Kan-PEG24-VAPAB-Exa” or “Kan- PEG24-VAPAB-E”.

[0388] Commercially available ADCs, such as trastuzumab deruxtecan (Dxd) (ENHERTU®), anti-TROP-2 sacituzumab govitecan, and other ADCs were also used as comparatives.

[0389] Comparative Formula ID 3080 was produced by conjugating nonspecific human IgG (HlgG) with mixed two biomolecules -SMCC-L(PEOX10-p- E and -Mal-PEG2-VC-MMAE, similar to Formula ID 3036. In one preparation, n=1 and n -3. In another preparation, n=1 and n’=4. In yet another preparation, n=4 and n’=1 . In yet another preparation, n=3 and n -1 .

Claims

CTCLAIMSWhat is claimed is:1 . A bioactive composition comprising a payload bioactive agent (PBA) comprising a formula selected from:a pharmaceutically acceptable salt thereof, a solvate thereof, a prodrug thereof, an isomer thereof, an isotope derivative thereof, and a combination thereof; wherein, v and w each is an integer, v=1-100; w=1-1000;CTR, R’, R”, R3, R4, R3’, R4’, Rs, Re, R7, Rs, and R15 each is independently H, D, C1 -C100 saturated or unsaturated hydrocarbon, C1 -C100 saturated or unsaturated hydrocarbon having at least one D, C1-C100 saturated or unsaturated hydrocarbon having at least one heteroatom, C1 -C100 saturated or unsaturated hydrocarbon having at least one heteroatom and at least one D, aromatic group, substituted aromatic group, or a combination thereof;R9 is selected from H, D, C1-C100 saturated or unsaturated hydrocarbon, C1 -C100 saturated or unsaturated hydrocarbon having at least one D, C1 -C100 saturated or unsaturated hydrocarbon having at least one heteroatom, C1-C100 saturated or unsaturated hydrocarbon having at least one heteroatom and at least one D, OH, NH2, Cl, F, I or Br;R10 is selected from H, Cl, F, I, Br and methoxy;R11 is OH, OR12, SH, SR13, NH2, NHD, ND2, or NRuR’u, wherein R12 is an alcohol protecting group, R13 is a thiol protecting group, and Ru and R’14 each is independently H, D, or an amine protecting group; said heteroatom is selected from N, O, S, P, Cl, Br, I and a combination thereof; andX is O, S, or NH.

2. The bioactive composition of claim 1 , wherein said PBA comprises a formula selected from Formula ID 1 , 1-1 , 1-2, 2, 2-1 , 2-2, 3, 3-1 , 3-2, 1001 ,1002, 1003, 1004, 1005, 1006, 1007, 1008, 1009, 1010, 1011 , 1012, 1013,1014, 1015, 1016, 1017, 1018, 1019, 1020, 1021 , 1022, 1023, 1024, 1025,1026, 1027, 1028, 1029, 1030, 1031 , 1032, 1033, 1034, 1035, 1036, 1037,1038, 10 1040, 1041 , 1042, 1043, 1044, 1045, 1046, 1047, 1048, 1049, 1050, 1051 , 1052, 1053, 1054, 1055, 1056, 1057, 1058, 1059, 1060, 1061 , 1062, 1063, 1064, 1065, 1066,1067, 1068, 1069, 1070, 1071 , 1072, 1073, 1074, 1075, 1076, 1077, 1078, 1079, 1080, 1081 , 1082, 1083, 1084, 1085, 1086,1087, 1088, 1089, 1090, 1091 , 1092, 1093, 1094, 1095, 1096, 1097, 1098,1099, 1100, 1101 , 1102, 1103, 1104, 1105, 1106, 1107, 1 108, 1109, 1110,1111 , 1112, 1113, 1114, 1115, 1116, 1117, 1118, 1119, 1 120, 1121 , 1122,1123, 1124, 1 125, 1126, 1127, 1128, 1129, 1130, 1131 , 1132, 1133, 1134,1135, a pharmaceutically acceptable salt thereof, a solvate thereof, a prodrugCT thereof, an isomer thereof, an isotope derivative thereof, and a combination thereof.

3. The bioactive composition of any one of claims 1-2 further comprising a polymer, a linker, optionally a coupler, or a combination thereof, wherein said PBA is covalently linked to said polymer, said linker, or optionally said coupler when present, wherein, said polymer comprises a peptide, a polyethylene glycol (PEG), polyoxazoline (POX), or a combination thereof, wherein said polyoxazoline (POX) comprises a linear portion, a branched portion, or a combination thereof, and wherein the polyoxazoline (POX) comprises poly(2-methyloxazoline), poly(2-ethyloxazoline), poly(2-propyloxazoline), poly(2-isopropyloxazoline), poly(2-substituted oxazoline), or a combination thereof.

4. The bioactive composition of any one of claims 1-3 comprising a formula selected from Formula ID 10, 11 , 12, 13, 14, 21 , 22, 23, 24, 25, 26, or a combination thereof.

5. The bioactive composition of claim 3 or 4, further comprising a polymer side chain that is covalently linked to said polymer, said linker, or a combination thereof, wherein said polymer side chain comprises POX comprising poly(2- methyloxazoline), poly(2-ethyloxazoline), poly(2-propyloxazoline), poly(isopropyloxazoline), or a combination thereof.

6. The bioactive composition of any one of claims 1-5, wherein said PBA further comprises calicheamicin, DM1 , DM4 (Ravtansine), Dxd (deruxtecan), exatecan, IRDye700DX, MMAE (Monomethyl auristatin E), vedotin, MMAF (Monomethyl auristatin F), Pseudomonas exotoxin A, pyrrolobenzodiazepine (PBD), SG3199 (PBD dimer), SN-38, irinotecan, camptothecin, amanitin, duocarmycins (DNA-alkylators), halichondrin, Eribulin, norhalichondrin B, maytansinoids, maitansine (INN), maytansine, PUN-159682, tubulysins, tubulysin, pretubulysin (PT), tubulysin A (TubA), tubulysin M, or a combination thereof.

7. The bioactive composition of any one of claims 3-6, wherein said linker comprises a cleavable linker selected from Formula ID 101 , 102, 103, 104, 105,CT106, 107, 108, 109, 110, 11 1 , 112, 1 13, 114, 115, 116, 117, 118, 119, 120, 121 , 122, 123, 124, 125, 126, 127, 128, 129, 130, 131 , 132, 133, 134, 135, 136, 137, 138, 139, 140, 141 , and a combination thereof.

8. The bioactive composition of any one of claims 3-6 comprising said coupler that comprises Formula ID 201 , 202, 203, 204, 205, 206, 207, 208, 209, or a combination thereof.

9. The bioactive composition of any one of claims 3-7, wherein said bioactive composition comprises a biomolecule having a formula selected from Formula ID 2001 , 2002, 2003, 2004, 2005, 2006, 2007, 2008, 2009, 2010, 2011 , 2012, 2013, 2014, 2015, 1026, 2017, 2018, 2019, 2020, 2021 , 2022, 2023, 2024, 2025, 2026, 2027, 2028, 2029, 2030, 2031 , 2032, 2033, 2034, 2035, 2036, 2037, 2038, 2039, 2040, a pharmaceutically acceptable salt thereof, a solvate thereof, a prodrug thereof, an isomer thereof, an isotope derivative thereof, and a combination thereof.

10. A pharmaceutical composition comprising a payload bioactive agent (PBA) comprising a formula selected from:CTa pharmaceutically acceptable salt thereof, a solvate thereof, a prodrug thereof, an isomer thereof, an isotope derivative thereof, a reacted PBA thereof, optionally an additional cytotoxic agent, and a combination thereof; and a pharmaceutical suitable carrier; wherein, v and w each is an integer, v=1-100; w=1-1000;R, R’, R”, R3, R4, R3’, R4’, Rs, Re, R7, Re, and R15 each is independently H, D, C1 -C100 saturated or unsaturated hydrocarbon, C1-C100 saturated or unsaturated hydrocarbon having at least one D, C1-C100 saturated or unsaturated hydrocarbon having at least one heteroatom, C1-C100 saturated or unsaturated hydrocarbon having at least one heteroatom and at least one D, aromatic group, substituted aromatic group, or a combination thereof;R9 is selected from H, D, C1-C100 saturated or unsaturated hydrocarbon, C1 -C100 saturated or unsaturated hydrocarbon having at least one D, C1 -C100 saturated or unsaturated hydrocarbon having at least one heteroatom, C1- C100 saturated or unsaturated hydrocarbon having at least one heteroatom and at least one D, OH, NH2, Cl, F, I, or Br;R10 is selected from H, Cl, F, I, Br, and methoxy;R11 is OH, OR12, SH, SR13, NH2, NHD, ND2, NR R’U, wherein R12 is an alcohol protecting group, R13 is a thiol protecting group, and Ri4 and R’14 each is independently H, D, or an amine protecting group; said heteroatom is selected from N, O, S, P, Cl, Br, I and a combination thereof; andX is O, S, or NH.11 . The pharmaceutical composition of claim 10, wherein said PBA comprises a formula selected from Formula ID 1 , 1-1 , 1-2, 2, 2-1 , 2-2, 3, 3-1 , 3-2, 1001 , 1002, 1003, 1004, 1005, 1006, 1007, 1008, 1009, 1010, 1011 , 1012, 1013,1014, 1015, 1016, 1017, 1018, 1019, 1020, 1021 , 1022, 1023, 1024, 1025,1026, 1027, 1028, 1029, 1030, 1031 , 1032, 1033, 1034, 1035, 1036, 1037,1038, 1039, 1040, 1041 , 1042, 1043, 1044, 1045, 1046, 1047, 1048, 1049,1050, 1051 , 1052, 1053, 1054, 1055, 1056, 1057, 1058, 1059, 1060, 1061 ,1062, 1063, 1064, 1065, 1066, 1067, 1068, 1069, 1070, 1071 , 1072, 1073,1074, 1075, 1076, 1077, 1078, 1079, 1080, 1081 , 1082, 1083, 1084, 1085,1086, 1087, 1088, 1089, 1090, 1091 , 1092, 1093, 1094, 1095, 1096, 1097,1098, 1099, 1 100, 1101 , 1102, 1103, 1104, 1105, 1106, 1 107, 1108, 1109,1110, 1111 , 1 112, 1113, 1114, 1115, 1116, 1117, 1118, 1 119, 1120, 1121 ,1122, 1123, 1 124, 1125, 1126, 1127, 1128, 1129, 1130, 1131 , 1132, 1133,1134, 1135, said additional cytotoxic agent when present comprises calicheamicin, DM1 , DM4 (Ravtansine), Dxd (deruxtecan), exatecan,IRDye700DX, MMAE (Monomethyl auristatin E), vedotin, MMAF (Monomethyl auristatin F), Pseudomonas exotoxin A, pyrrolobenzodiazepine (PBD), SG3199 (PBD dimer), SN-38, irinotecan, camptothecin, amanitin, duocarmycins (DNA- alkylators), halichondrin, Eribulin, norhalichondrin B, maytansinoids, maitansine (INN), maytansine, PUN-159682, tubulysins, tubulysin, pretubulysin (PT), tubulysin A (TubA), tubulysin M, a pharmaceutically acceptable salt thereof, a solvate thereof, a prodrug thereof, an isomer thereof, an isotope derivative thereof, or a combination thereof.

12. The pharmaceutical composition of claim 10 or 11 further comprising: a targeting bioactive agent (TBA) comprising binding affinity to a target molecule or a target cell, wherein said PBA is reacted and covalently linked to said TBA directly or indirectly.

13. The pharmaceutical composition of any one of claims 10-12 further comprising a polymer, a linker, optionally a coupler, or a combination thereof, wherein said PBA is covalently linked to said polymer, said linker, or optionally, said coupler when present, or a combination thereof, wherein,CT said polymer comprises a peptide, a polyethylene glycol (PEG), polyoxazoline (POX), or a combination thereof, wherein said polyoxazoline (POX) comprises a linear portion, a branched portion, or a combination thereof, and wherein the polyoxazoline (POX) comprises poly(2-methyloxazoline), poly(2-ethyloxazoline), poly(2-propyloxazoline), poly(2-isopropyloxazoline), poly(2-substituted oxazoline), or a combination thereof.

14. The pharmaceutical composition of claim 13, wherein said linker comprises a Linker L1 , a Linker L2, or a combination thereof, wherein said Linker L1 is covalently linked to said TBA and covalently linked to a reacted first terminal group of said polymer, said Linker L2 is covalently linked to said PBA and covalently linked to a reacted second terminal group of said polymer.

15. The pharmaceutical composition of claim 14, wherein at least one of said Linker L1 and said Linker L2 comprises a cleavable linker that is cleavable in vivo or in the target molecule or the target cell, and wherein said cleavable linker comprises an enzymatically-cleavable linker, acid cleavable linker, acid sensitive hydrazone linker, glutathione (GSH) cleavable linker, a linker comprising disulfides, SPDB glutathione cleavable linker, thioether linker, glutathione-sensitive disulfide linker, Fe(ll) cleavable linker, cathepsin cleavable linker, a linker containing a sugar molecule or moiety, glucuronide containing linker, glycosidase cleavable linker, phosphatase cleavable linker, sulfatase cleavable linker, esterase cleavable linker, hydrolysis cleavable linker, Photo-responsive cleavable linker, Bioorthogonal cleavable linker, tumor microenvironment activatable linker, Ortho Hydroxy-Protected Aryl Sulfate (OHPAS) linker, modified monodispersed PEG linker, modified PEO linker, modified poly-PEG linker, enzymatically-cleavable peptide linker, dipeptide linker, valine— citrulline linker (VC linker), valine-alanine linker, VAPAB linker (Valine-alanine-p-aminobenzyl), L-Val-L-Ala-para-aminobenzyl alcohol (VAPAB-OH) linker, VCPAB (Valine-citrulline-p-aminobenzyl) linker, L-Val-L- Cit-para-aminobenzyl alcohol (VCPAB-OH) linker, polypeptide linker, Gly-Gly- L-Phe-N-[(carboxymethoxy)methyl]Glycinamid (GGFG-NCM) linker, GGFG (Glycyl glycyl-L-phenylalanyl glycyl) linker, glutamic acid-glycine-citrullineCT(EGCit, or Glu-Gly-Cit) linker, tripeptide linker, glutamic acid-valine-citrulline (Glu-Val-Cit) linker, p-aminobenzyloxycarbonyl (PABC) linker, p- aminobenzyloxycarbonyl (PABC) linker, or a combination thereof.

16. The pharmaceutical composition of any one of claims 12-15, wherein said pharmaceutical composition comprises a formula selected from Formula ID 30, 31 , 32, 33, 34, 35, 36, 37, 38, 39, 40, or a combination thereof.

17. The pharmaceutical composition of any one of claims 13-16, comprising said coupler, wherein the coupler comprises Formula ID 201 , 202, 203, 204, 205, 206, 207, 208, 209, or a combination thereof.

18. The pharmaceutical composition of any one of claims 12-17, wherein the TBA is selected from a peptide; a protein; DNA; RNA; nucleic acids; oligo nucleic acids; mRNA; siRNA; sgRNA; an antibody selected from a monoclonal antibody, a fragment of a monoclonal antibody, a polyclonal antibody, a fragment of a polyclonal antibody, a synthetic antibody, a fragment of a synthetic antibody, antigen-binding portions thereof, and a combination thereof; an agonist; an activator; an inhibitor; an antagonist; a ligand of a cell surface receptor; a naked nucleic acid; a nucleic acid; an antisense nucleic acid, anti- CD19; a ribozyme; a virus; a virus-like particles and a combination thereof; chimeric antigen receptor or chimeric antigen T-cell receptor (CAR-T), cytokines; granulocyte colony stimulating factor (G-CSF); interferon a; interferon a2a; interleukins; lectins; Neupogen (Filgrastim); siRNAs; T-cell receptor (TCR); a member of a small molecule binding pair and a combination thereof; and a combination of any of the foregoing.

19. The pharmaceutical composition of any one of claims 12-18, further comprising a polymer side chain that is covalently linked to said TBA, said polymer, said linker, or a combination thereof, wherein said polymer side chain comprises POX comprising poly(2-methyloxazoline), poly(2-ethyloxazoline), poly(2-propyloxazoline), poly(isopropyloxazoline), lysine, arginine, glutamic acid, glutamine, a reacted form thereof, a derivative thereof, or a combination thereof.CT20. The pharmaceutical composition of claim 18 or 19, wherein said TBA comprises said antibody having binding affinity to said target molecule that comprises a tumor antigen (Ag) and the pharmaceutical composition comprises tumor cytotoxicity comprising AgHightumor cytotoxicity and AgLowtumor cytotoxicity.21 . The pharmaceutical composition of any one of claims 19-20, wherein said antibody (Ab) comprises anti-CD19 antibodies, anti-CD20 antibodies, anti- CTLA-4 (cytotoxic T lymphocyte-associated antigen) antibodies, anti-LAG3 (lymphocyte activation gene-3) antibodies, anti-PD1 antibodies (pembrolizumab), anti-PD-L1 antibodies, anti-TIM-3 (T cell immunoglobulin and mucin domain-3) antibodies, one or more antibodies against target molecules comprising 5T4, ALPPL2, AXL, B7H3, B7H4, B7H6, B7S1 , B7x, BCAM, BCMA, CCL2, CCL5, CD146, CD25, CD27 ligand, CD276, CD47, CD70, CDH3, CDH6, CEACAM5, CEACAM6, CLDN1 , CLDN18.2, CLDN6, DLK1 , DLL3, EGFR, EGFRVIII, EpCAM, ErbB1 , ErbB2, ErbB3, FAT2, FOLR1 , GPC3, GPR87, GRM8, GRP56, HER2, HER3, HGFR, ITGB6, LIV-1 , Ly6G6D, LY6G6F, LYPD3, MCAM, MET, MSLN, MUC16, MUC18, PSMA, ROR1 , SEZ6, SLC39A6, SLC7A11 , SLCO1 B3, STEAP1 , Tacstd2, TLR4, TMPRSS4, TROP2, UFO, VEGF, VTCN1 , ZIP6, a bispecific antibody thereof, a tri-specific antibody thereof, a tetra-specific antibody thereof, a fragment thereof, an antigen-binding portion thereof, or a combination thereof.

22. The pharmaceutical composition of any one of claims 19-20, wherein said antibody (Ab) comprises alemtuzumab, atezolizumab, avelumab, bevacizumab, blinatumomab, brentuximab, cetuximab, daratumumab, datopotamab, denosumab, dinutuximab, disitamab, durvalumab, elotuzumab, enfortumab, gemtuzumab, ibritumomab, durvalumab, inotuzumab, ipilimumab, lectins, loncastuximab, mirvetuximab, necitumumab, obinutuzumab, ofatumumab, olaratumab, nivolumab, panitumumab, pembrolizumab, pertuzumab (anti-HER2), polatuzumab, ramucirumab, rovalpituzumab, rituximab, rituximab, sacituzumab, siltuximab, toripalimab, tislelizumab (BGB- A317), tisotumab, atezolizumab, tositumomab, trastuzumab (anti-HER2), or a combination thereof.

23. The pharmaceutical composition of either claim 21 or 22, wherein said Ab is covalently linked to said PBA, said coupler, said polymer, said linker, said Linker L1 , said Linker L2, or a combination thereof, to form an antibody-drug conjugate (ADC) having a formula selected from Formula ID 50, 51 , 52, 53, 54, 55, 56, 57, 58, 59, 60, 61 , 62, 63, 64, 65, and a combination thereof.

24. The pharmaceutical composition of claim 23, wherein said Ab is covalently linked to one or more same or different biomolecules having a formula selected from Formula ID 2001 , 2002, 2003, 2004, 2005, 2006, 2007, 2008, 2009, 2010, 2011 , 2012, 2013, 2014, 2015, 1026, 2017, 2018, 2019, 2020, 2021 , 2022, 2023, 2024, 2025, 2026, 2027, 2028, 2029, 2030, 2031 , 2032, 2033, 2034, 2035, 2036, 2037, 2038, 2039, 2040, a pharmaceutically acceptable salt thereof, a solvate thereof, a prodrug thereof, an isomer thereof, an isotope derivative thereof, and a combination thereof, to form said antibody-drug conjugate (ADC).

25. The pharmaceutical composition of any one of claims 12-24, comprising two or more of said PBAs.

26. The pharmaceutical composition of claim 25, wherein said two or more PBAs comprise Formula ID 1 , 1-1 , 1-2, 2, 2-1 , 2-2, 3, 3-1 , 3-2, 1001-1135, exatecan, deruxtecan, MMAE, MMAF, a pharmaceutically acceptable salt thereof, a solvate thereof, a prodrug thereof, an isomer thereof, an isotope derivative thereof, or a combination thereof.

27. The pharmaceutical composition of any one of claims 12-26, comprising a Formula ID 3001-3074, or a combination thereof.

28. The pharmaceutical composition of any one of claims 12-27, comprising a TBA to PBA ratio in a range of from 1 : 1 to 1 :24.

29. A pharmaceutical composition comprising a Nano Antibody-drug Conjugate (Nano ADC) comprising a payload bioactive agent (PBA) comprising a formula selected from:a pharmaceutically acceptable salt thereof, a solvate thereof, a prodrug thereof, an isomer thereof, an isotope derivative thereof, a reacted PBA thereof, optionally an additional cytotoxic agent, and a combination thereof; and an antibody selected from a monoclonal antibody (mAb) HER2 receptor antagonist trastuzumab biosimilar, an anti-human B7-H3 (CD276) monoclonal antibody, or a non-specific human IgG (HlgG); optionally, a pharmaceutical suitable carrier; wherein, v and w each is an integer, v=1-100; w=1-1000;R, R’, R”, R3, R4, R3’, R4’, R5, R6, R7, R8, and R15 each is independently H, D, C1 -C100 saturated or unsaturated hydrocarbon, C1 -C100 saturated or unsaturated hydrocarbon having at least one D, C1-C100 saturated or unsaturated hydrocarbon having at least one heteroatom, C1-C100 saturated or unsaturated hydrocarbon having at least one heteroatom and at least one D, aromatic group, substituted aromatic group, or a combination thereof;R9 is selected from H, D, C1-C100 saturated or unsaturated hydrocarbon, C1 -C100 saturated or unsaturated hydrocarbon having at least one D, C1 -C100 saturated or unsaturated hydrocarbon having at least one heteroatom, C1- C100 saturated or unsaturated hydrocarbon having at least one heteroatom and at least one D, OH, NH2, Cl, F, I, or Br;R10 is selected from H, Cl, F, I, Br, and methoxy;R11 is OH, OR12, SH, SR13, NH2, NHD, ND2, NRuR’u, wherein R12 is an alcohol protecting group, R13 is a thiol protecting group, and Ru and R’14 each is independently H, D, or an amine protecting group; said heteroatom is selected from N, O, S, P, Cl, Br, I and a combination thereof; andX is O, S, or NH.

30. A method for treating or preventing a disease of a subject in need thereof, the method comprising administering an effective dose of the pharmaceutical composition of any one of claims 10-29 to the subject, wherein the disease comprises prostate cancer, breast cancer, gastric cancer, colon cancer, small cell lung cancer, or colorectal adenocarcinoma.

31. Use of the pharmaceutical composition of any one of claims 10-29, for manufacturing of a medicament for treating or preventing a disease in a subject in need thereof, wherein the disease comprises prostate cancer, breast cancer, gastric cancer, colon cancer, small cell lung cancer, or colorectal adenocarcinoma.

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