Drug conjugate

Abstract

To provide antibody-drug conjugates.SOLUTION: The invention relates to antibody-drug conjugates (ADCs) that are potentiated by concomitant administration of a moiety comprising covalently-linked saponin. The invention also relates to antibody-oligonucleotide conjugates (AOCs) that are potentiated by concomitant administration of a moiety comprising covalently linked saponin. The invention also relates to ADCs and AOCs which are conjugated with a saponin via a covalent linker. The invention further relates to an effector moiety such as a toxin or an antisense oligonucleotide such as a BNA, conjugated with a saponin via a covalent linkage. The invention also relates to a BNA covalently conjugated with a targeting moiety such as an antibody. The invention also relates to combinations of therapeutic agents, comprising a first pharmaceutical composition comprising a conjugate of a cell-targeting moiety such as an antibody and an antisense oligonucleotide such as a BNA covalently bound thereto, and comprising a second pharmaceutical composition comprising either a free saponin, or a conjugate of a cell-targeting moiety such as an antibody with a saponin covalently linked thereto. Furthermore, the invention relates to any of these conjugates or therapeutic compositions or therapeutic combinations, for use as a medicament. The invention also relates to any of these conjugates or therapeutic compositions or therapeutic combinations, for use in a method for treatment or prophylaxis of a cancer. Finally, the invention relates to methods for producing any of these conjugates of the invention.SELECTED DRAWING: None

Description

[Technical Field]

[0001] The present invention relates to antibody-drug conjugates (ADCs) that are enhanced by coadministration of the ADC with a moiety comprising a covalently linked saponin. The present invention also relates to antibody-oligonucleotide conjugates (AOCs) that are enhanced by coadministration of the ADC with a moiety comprising a covalently linked saponin. The present invention also relates to ADCs and AOCs that are conjugated to saponin via a covalent linker. The present invention further relates to effector moieties, such as toxins or antisense oligonucleotides, e.g., BNAs, conjugated to saponin via a covalent linker. The present invention also relates to BNAs covalently conjugated to a targeting moiety, such as an antibody. The present invention also relates to therapeutic combinations, including a first pharmaceutical composition comprising a conjugate of a cell-targeting moiety, such as an antibody, and an antisense oligonucleotide, such as a BNA, covalently linked thereto, and a second pharmaceutical composition comprising either free saponin or a conjugate of a cell-targeting moiety, such as an antibody, with a saponin covalently linked thereto. Furthermore, the present invention relates to any of these conjugates or therapeutic compositions or therapeutic combinations for use as pharmaceuticals. The present invention also relates to any of these conjugates or therapeutic compositions or therapeutic combinations for use in methods for the treatment or prevention of cancer. Finally, the present invention relates to methods for producing any of these conjugates of the present invention. [Background technology]

[0002] Molecules with therapeutic biological activity are often theoretically suitable for application as effective therapeutic drugs for the treatment of diseases such as cancer in human patients in need thereof. A typical example is a small molecule biologically active moiety. However, many, if not all, potential drug-like molecules and therapeutic agents currently in clinical use suffer from at least one of a plethora of shortcomings and drawbacks. When administered to the human body, therapeutically active molecules may exert off-target effects in addition to biological activities directed at aspects underlying the disease or health problem to be treated. Such off-target effects are undesirable and carry the risk of inducing health- or even life-threatening side effects of the administered molecule. It is the occurrence of such adverse events that causes many drug-like compounds and therapeutic agent moieties to fail Phase III or even Phase IV clinical trials (post-marketing follow-up). Thus, there is a strong desire to provide drug molecules, such as small molecule therapeutics, whose therapeutic effect should, for example, be, among other things, (1) highly specific for the biological factors or processes driving the disease, (2) sufficiently safe, (3) sufficiently effective, (4) sufficiently directed at diseased cells with little or no off-target activity against non-diseased cells, (5) have a sufficiently timely mechanism of action (e.g., the administered drug molecule should reach the target site in a human patient within a certain time frame and remain at the target site for a certain time frame), and / or (6) have a sufficiently long-lasting therapeutic activity in the patient's body. Unfortunately, despite already long and intensive research, and despite impressive progress made in several areas where the challenges and shortcomings encountered have been individually addressed, to date, an "ideal" therapeutic agent possessing many or even all of the beneficial characteristics outlined herein above has not been available to patients.

[0003] Chemotherapy is one of the most important therapeutic options for cancer treatment. However, because it lacks specificity for cancer cells compared to dividing cells in healthy tissues, it is often associated with a narrow therapeutic window. The development of monoclonal antibodies has offered the possibility of exploiting their specific binding properties as a mechanism for targeted delivery of cytotoxic drugs to cancer cells while sparing normal cells. This can be achieved by chemically conjugating a cytotoxic effector (also known as a payload or warhead) to the antibody to create an antibody-drug conjugate (ADC). Typically, highly potent payloads such as emtansine (DM1), which has a limited therapeutic index (the ratio of the toxic dose to the effective dose) in its unconjugated form, are used. Conjugation of DM1 to trastuzumab (ado-trastuzumab-emtansine), also known as Kadcycla, improves the tolerable dose of DM1 by at least two-fold in monkeys. Over the past few decades, enormous effort and investment has been devoted to developing therapeutic ADCs. Despite promising preclinical data, however, bringing ADCs to the clinic remains challenging. The first ADC approved for clinical use was gemtuzumab ozogamicin (CD33-targeted Mylotarg, Pfizer / Wyeth) for relapsed acute myeloid leukemia (AML) in 2000. However, Mylotarg was withdrawn from the market at the request of the Federal Drug Administration (FDA) due to several concerns regarding its safety profile. Patients treated with Mylotarg were found to die more often than patients treated with conventional chemotherapy. Mylotarg was reapproved in 2017 with a lower recommended dose, a different schedule in combination with chemotherapy or on its own, and a new patient population. To date, only five ADCs have been approved for clinical use, while approximately 55 ADCs have been discontinued in clinical development. However, interest remains high, and approximately 80 ADCs are currently in clinical development with nearly 600 trials.

[0004] Despite the potential for using toxic payloads that are usually not tolerated by patients, a low therapeutic index (the ratio of toxic dose to efficacious dose) is a major issue that explains the discontinuation of many ADCs in clinical development. This can be caused by several mechanisms, such as off-target toxicity against normal cells, the development of resistance to cytotoxic drugs, and premature release of the drug in the circulation. A systematic review of ADCs by the FDA found that the toxicity profiles of most ADCs can be categorized according to the payload used rather than the antibody used, suggesting that toxicity is largely determined by premature release of the payload. It is estimated that of the approximately 55 ADCs that have been discontinued, at least 23 were due to a poor therapeutic index. For example, development of a trastuzumab-tesirin conjugate (HER-2-targeting ADCT-502, an ADC therapeutic) was recently halted due to a narrow therapeutic index, likely due to on-target normal tissue effects in lung tissue, which expresses significant levels of HER2. In addition, several ADCs in Phase 3 trials have been discontinued due to missing primary endpoints. For example, phase 3 trials of a depatuxizumab mafodotin conjugate (EGFR-targeting ABT-414, AbbVie) tested in patients with newly diagnosed glioblastoma and a mirvetuximab soravtansine conjugate (folate receptor alpha (FRα)-targeting IMGN853, ImmunoGen) tested in patients with platinum-resistant ovarian cancer were recently halted and failed to demonstrate survival benefit. It is important to note that the clinically used dose of some ADCs may not be sufficient for their full anticancer activity. For example, ado-trastuzumab emtansine has an MTD of 3.6 mg / kg in humans. In preclinical models of breast cancer, ado-trastuzumab emtansine induced tumor regression at dose levels of 3 mg / kg or higher, but more potent efficacy was observed at 15 mg / kg. This suggests that at clinically administered doses, ado-trastuzumab emtansine may not exert its maximum potential antitumor effect.

[0005] ADCs mainly consist of an antibody, a cytotoxic moiety such as a payload, and a linker. To overcome existing problems, several novel strategies have been proposed and implemented to target each component of an ADC. For example, by identifying and validating a satisfactory antigenic target for the antibody component, selecting an antigen that has a high expression level in tumors and no or little expression in normal tissues, an antigen that is present on the cell surface so as to be accessible to circulating ADCs, and an antigen that allows ADCs to be internalized into cells after binding; and alternative mechanisms of activity; designing and optimizing linkers that improve the solubility and drug-to-antibody ratio (DAR) of ADCs and overcome resistance induced by proteins that can transport chemotherapeutic drugs out of cells; and selecting and optimizing antibodies to improve the DAR ratio by incorporating more payload and improve the homogeneity and developability of antibodies. In addition to ADC technology developments, new clinical and translational strategies are also being developed to maximize the therapeutic index, altering dosing schedules, for example with split dosing; conducting biodistribution studies; and incorporating biomarkers to optimize patient selection, capture response signals early, monitor the duration and depth of response, and inform combination studies.

[0006] Examples of ADCs with clinical potential include those being evaluated as treatment options for lymphoid malignancies and multiple myeloma, such as brentuximab vedotin, inotuzumab ozogamicin, moxetumomab pasudotox, and polatuzumab vedotin. Polatuzumab vedotin, which binds to CD79b on (malignant) B cells, and pinatuzumab vedotin, which binds to CD22, are being tested in clinical trials. Each ADC was combined with rituximab, a monoclonal antibody that binds to CD20, and no payload was provided (B. Yu and D. Liu, Antibody-drug conjugates in clinical trials for lymphoid malignancies and multiple myeloma; Journal of Hematology & Oncology (2019) 12:94). Combinations of monoclonal antibodies, such as these examples, are yet another approach, attempting to arrive at a "magic bullet" that combines many or even all of the previously mentioned desirable characteristics of ADCs.

[0007] Meanwhile, nucleic acid-based therapeutics have been under development for the past few decades. Therapeutic nucleic acids can be based on deoxyribonucleic acid (DNA) or ribonucleic acid (RNA), antisense oligonucleotides (ASOs, AONs), and short interfering RNA (siRNA), microRNA, and DNA and RNA aptamers for approaches such as gene therapy, RNA interference (RNAi). Many of them share the same fundamental basis of action: inhibiting either DNA or RNA expression, thereby preventing the expression of abnormal proteins associated with disease. The largest number of clinical trials are in the field of gene therapy, with nearly 2,600 ongoing or completed trials worldwide, but only about 4% have entered phase 3. This is followed by ASO-based clinical trials. Similar to ADCs, despite the numerous technologies being explored, therapeutic nucleic acids share two major issues during clinical development: intracellular delivery and off-target effects. ASOs, such as peptide nucleic acids (PNAs), phosphoramidate morpholino oligomers (PMOs), locked nucleic acids (LNAs), and bridged nucleic acids (BNAs), are being explored as attractive strategies for specifically inhibiting target genes, particularly those that are difficult to target with small molecule inhibitors or neutralizing antibodies. The efficacy of different ASOs is currently being investigated in many neurodegenerative diseases, such as Huntington's disease, Parkinson's disease, Alzheimer's disease, and amyotrophic lateral sclerosis, as well as in several cancer stages. The application of ASOs as potential therapeutic agents requires safe and effective methods for their delivery to the cytoplasm and / or nucleus of target cells and tissues. Although the clinical relevance of ASOs has been demonstrated, inefficient cellular uptake both in vitro and in vivo has limited their efficacy and been a barrier to their therapeutic development. Cellular uptake can be <2% of the dose, resulting in ASO concentrations at the active site that are too low for effective and sustained outcomes. This consequently requires increased administration doses, which induce off-target effects. The most common side effects are activation of the complement cascade, inhibition of the coagulation cascade, and toll-like receptor-mediated stimulation of the immune system.

[0008] Chemotherapeutic drugs are most commonly small molecules. However, their effectiveness is hindered by severe secondary off-target toxicity, as well as their poor solubility, rapid clearance, and limited tumor exposure. Scaffold-small molecule drug conjugates, such as polymer-drug conjugates (PDCs), are pharmacologically active polymeric constructs that contain one or more molecules of a small molecule drug bound to a carrier scaffold (e.g., polyethylene glycol (PEG)).

[0009] This conjugate principle has attracted considerable attention and has been under investigation for decades. The majority of small molecule drug conjugates in preclinical or clinical development are for oncology indications. However, to date, only one non-cancer drug (Movantik, AstraZeneca, a PEG oligomer conjugate of the opioid antagonist naloxone) has been approved for opioid-induced constipation in chronic pain patients in 2014. This is a non-oncology indication. The translational application of drug-scaffold conjugates to the treatment of human subjects has so far met with little clinical success. For example, PK1 (N-(2-hydroxypropyl)methacrylamide (HPMA) copolymer doxorubicin; developed by Pharmacia, Pfizer) has shown significant anticancer activity in both solid tumors and leukemia in mouse models and is under clinical investigation for oncology indications. Although it demonstrated significantly reduced non-specific toxicity and improved pharmacokinetics in humans, the improvement in anti-cancer efficacy proved to be modest in patients, and as a result, further development of PK1 was discontinued.

[0010] The failure of scaffold-small molecule drug conjugates has been attributed, at least in part, to their poor accumulation at tumor sites. For example, in murine models, PK1 showed 45-250-fold higher accumulation in tumors than in healthy tissues (liver, kidney, lung, spleen, and heart), although tumor accumulation was only observed in a small subset of patients in clinical trials.

[0011] A potential solution to the previously mentioned problems is the application of nanoparticle systems for drug delivery, such as liposomes. Liposomes are spherical vesicles composed of one or more phospholipid bilayers that spontaneously form when phospholipids are dispersed in water. The amphiphilic properties of phospholipids provide them with properties of self-assembly, emulsification, and wetting characteristics, which can be used to design new drugs and new drug delivery systems. Drugs encapsulated in liposomal delivery systems may offer several advantages over direct drug administration, such as improved and controlled pharmacokinetics and pharmacodynamics, tissue targeting, reduced toxicity, and enhanced drug activity. One such successful example is the liposomal form of the small molecule chemotherapy drug doxorubicin (Doxil: pegylated liposomal form of doxorubicin; Myocet: non-pegylated liposomal doxorubicin), which has been approved for clinical use.

[0012] Thus, there remains a need to find solutions that allow drug treatments, such as antitumor treatments, to be applicable for non-systemic use when desired, with drugs that have, for example, an acceptable safety profile, little off-target activity, sufficient efficacy, a sufficiently low clearance rate from the patient's body, etc. Summary of the Invention

[0013] In some embodiments of the present invention, it is a primary goal to provide improved bioactive compounds or compositions containing such improved bioactive compounds.

[0014] It is one of several objects of embodiments of the present invention to provide a solution to the problem of non-specificity faced when administering small molecule therapeutically active compounds to human patients in need thereof. It is one of several objects of embodiments of the present invention to provide a solution to the problem of drugs having suboptimal specificity for disease-driving biological factors or biological processes. It is one of several objects of embodiments of the present invention to provide a solution to the problem of inadequate safety characteristics of current drugs when administered to human patients in need thereof. It is one of several objects of embodiments of the present invention to provide a solution to the problem of current drugs being less effective than desired when administered to human patients in need thereof. It is one of several objects of embodiments of the present invention to provide a solution to the problem of current drugs not being adequately targeted to diseased cells with little or no off-target activity against non-diseased cells when administered to human patients in need thereof. It is one of several objects of embodiments of the present invention to provide a solution to the problem that current drugs, when administered to a human patient in need thereof, do not have a sufficiently timely mechanism of action (e.g., the administered drug molecule should reach the target site in the human patient within a certain time frame and should remain at the target site for a certain time frame).It is one of several objects of embodiments of the present invention to provide a solution to the problem that current drugs, when administered to a human patient in need thereof, do not have a sufficiently long-lasting therapeutic activity in the patient's body.

[0015] At least one of the above objects of the present invention is achieved by providing a conjugate of the present invention, preferably comprising a cell-targeting moiety and at least one saponin. The conjugate is also suitable for use as a pharmaceutical or for involvement in a pharmaceutical combination according to the present invention, and for use as a semi-finished product in the manufacture of an ADC or antibody-oligonucleotide conjugate (AOC) of the present invention according to the present invention. The therapeutic drug combination, for example, comprises a conjugate of the present invention comprising a covalently attached saponin, and a second conjugate comprising, for example, an effector molecule, also referred to as an effector moiety or payload, wherein the conjugates comprise different binding sites for different epitopes exposed on different cell surface molecules of a target cell, the different cell surface molecules being expressed by and exposed on the surface of the same target cell, or the conjugates comprise the same binding site for the same epitope on the same cell surface molecule of the target cell.

[0016] While the present invention will be described with reference to specific embodiments, the invention is not limited thereto but only by the claims. The embodiments of the invention described herein can work in combination and in concert, unless otherwise specified.

[0017] One aspect of the present invention relates to a conjugate, comprising or consisting of an antibody and an antisense oligonucleotide, e.g., an antisense BNA, covalently linked together. Figures 1-5 illustrate the gene silencing activity of such a conjugate (in vivo testing in animal tumor models). Reference is also made to the Examples section.

[0018] One aspect of the present invention relates to the combination of a first composition comprising a conjugate comprising or consisting of an antisense oligonucleotide, such as an antisense BNA, and an antibody covalently linked together, and a second composition comprising a free saponin of the present invention (see Table A1, Scheme I). The gene silencing activity of such a conjugate is illustrated in Figures 1-7A and 1-7C (in vitro cell-based bioassay with human tumor cells). Reference is also made to the Examples section.

[0019] Some embodiments of the present invention also relate to pharmaceutical combinations, comprising or consisting of a first composition comprising a first conjugate comprising or consisting of an antibody and an antisense oligonucleotide, such as an antisense BNA, and a second composition comprising a first conjugate comprising or consisting of the same antibody and at least one saponin of the present invention. Figures 1-5 illustrate the gene silencing activity of such conjugates (in vivo testing in animal tumor models). Figures 8-5 illustrate the gene silencing activity of such conjugates (in vitro cell-based bioassay using human tumor cells). Reference is also made to the Examples section.

[0020] One embodiment of the present invention relates to a pharmaceutical combination, comprising or consisting of a fourth composition comprising a fourth conjugate comprising or consisting of an antibody and an antisense oligonucleotide, such as an antisense BNA, and a fifth composition comprising a first conjugate comprising or consisting of a different antibody and at least one saponin of the present invention. Figures 10-6A and 10-6C illustrate the gene silencing activity of such conjugates (in vitro cell-based bioassay using human tumor cells). Reference is also made to the Examples section.

[0021] One aspect of the present invention relates to conjugates comprising or consisting of an antisense oligonucleotide, such as an antisense BNA, covalently linked to at least one saponin of the present invention. Figures 1-3 illustrate the gene silencing activity of such conjugates (in vitro cell-based bioassay with human tumor cells). Reference is also made to the Examples section.

[0022] One embodiment of the present invention relates to a conjugate comprising or consisting of an antisense oligonucleotide, such as an antisense BNA, covalently coupled to a polymer backbone, such as a dendron, such as a G4-dendron, where the polymer backbone is covalently conjugated to one or more saponin molecules of the present invention, such as four saponin molecules. Figures 1-3 illustrate the gene silencing activity of such conjugates (in vitro cell-based bioassay with human tumor cells). Reference is also made to the Examples section.

[0023] One embodiment of the present invention relates to a conjugate comprising or consisting of an antibody, such as a monoclonal antibody, having specificity for a tumor marker or tumor cell receptor, covalently linked to at least one antisense oligonucleotide molecule, such as an antisense BNA, and covalently linked to at least one saponin molecule of the present invention. Figures 2-4 illustrate the gene silencing activity of such conjugates (in vivo testing in animal tumor models). Reference is also made to the Examples section.

[0024] One embodiment of the present invention relates to a conjugate comprising or consisting of an antibody, such as a monoclonal antibody, having specificity for a tumor marker or tumor cell receptor, covalently linked to at least one antisense oligonucleotide molecule, such as an antisense BNA, via a trifunctional linker, such as the linker of Scheme II, and covalently linked to at least one saponin molecule of the present invention via the same trifunctional linker. Figure 1-1 illustrates the gene silencing activity of such a conjugate (in vivo testing in an animal tumor model). Reference is also made to the Examples section.

[0025] One embodiment of the present invention relates to a combination of therapeutic agents, including an eighth composition comprising or consisting of a conjugate comprising or consisting of an antibody, preferably a monoclonal antibody with specificity for a tumor marker or tumor cell receptor, covalently linked to at least one saponin molecule of the present invention, preferably via at least one linker, preferably at least one cleavable linker cleavable under physiological acidic conditions, and a ninth composition further comprising an antisense oligonucleotide, such as an antisense BNA molecule. Figure 6-2 illustrates the gene silencing activity of such a conjugate (in vivo testing in an animal tumor model). Figures 5-2A and 5-2C illustrate the gene silencing activity of such a conjugate (in vitro cell-based bioassay with human tumor cells). Reference is also made to the Examples section.

[0026] One aspect of the invention relates to any of the previously mentioned conjugates or compositions or therapeutic combinations for use as a medicament.

[0027] One aspect of the invention relates to any of the previously mentioned conjugates or compositions or therapeutic combinations for use in the treatment or prevention of cancer.

[0028] One aspect of the present invention relates to a therapeutic drug molecule having the chemical structure of Compound I: A1m ((-L9 w )((-L1 q -B1 n ) u ((-L2 r -L3 s )(-L4 v -C) p ) t )) x

[0029] (Compound I) During the ceremony, A1 is a first ligand when B1 is a first effector moiety, or A1 is a first effector moiety when B1 is a first ligand; C is saponin; When A1 is a first ligand and B1 is a first effector moiety, then m=0 or 1; When A1 is a first effector moiety and B1 is a first ligand, m=0 to 32; When B1 is a first ligand and A1 is a first effector moiety, or when A1 is a first ligand and B1 is a first effector moiety, n=0 or 1; p = 1 to 128; L1 is at least one linker for covalently coupling two chemical groups; L2 is at least one linker for covalently coupling two chemical groups; L3 is at least one oligomeric or polymeric backbone for covalently coupling two chemical groups; L4 is at least one linker for covalently coupling two chemical groups; L9 is a trifunctional linker for covalently coupling three chemical groups; q=0 or 1; r=0 or 1; s=0 or 1; If s=0, t=0, 1, or 2, and if s=1, t=0 to 16; u=0-32 when A1 is a first ligand and B1 is a first effector moiety, or u=1 when A1 is a first effector moiety and B1 is a first ligand; v=0 or 1; w=1 or 0; x=1~16.

[0030] One embodiment relates to a therapeutic combination, comprising a therapeutic molecule according to the invention and a second therapeutic molecule having the chemical structure of Compound II: A2 a ((-L10 j )((-L5 d -B2 b ) h ((-L6 e -L7 f )(-L8 i -C) c ) g )) k

[0031] (Compound II) During the ceremony, A2 is a second ligand when B2 is a second effector moiety, or A2 is a second effector moiety when B2 is a second ligand; C is saponin; a=0 or 1 when A2 is a second ligand and B2 is a second effector moiety, or a=0-32 when A2 is a second effector moiety and B2 is a second ligand; b=0 or 1 when B2 is a second ligand and A2 is a second effector moiety, or when A2 is a second ligand and B2 is a second effector moiety; c = 1 to 128; L5 is at least one linker for covalently coupling two chemical groups; L6 is at least one linker for covalently coupling two chemical groups; L7 is at least one oligomeric or polymeric backbone for covalently coupling two chemical groups; L8 is at least one linker for covalently coupling two chemical groups; L10 is a trifunctional linker for covalently coupling three chemical groups; d=0 or 1; e=0 or 1; f=0 or 1; When f=0, g=0, 1, 2, when f=1, g=0 to 16; h=0-32 when A2 is a second ligand and B2 is a second effector moiety, or h=1 when A2 is a second effector moiety and B2 is a second ligand; i=0 or 1; j=1 or 0; k=1 to 16.

[0032] One embodiment relates to a therapeutic molecule of the invention, wherein r=0, s=0, v=0, s=0, v=0, p=0, t=0, the ligand A1 is a monoclonal antibody or at least one binding fragment or domain thereof according to any one of claims 3 to 7, m=1, the effector moiety B1 is an effector moiety according to claim 8, preferably a BNA, q=0, n=1, and u=2 to 4, or either q=1, n=2 to 4, u=2 to 4, and the linker L1 is an oligomeric or polymeric scaffold L3 according to any one of claims 21 to 30.

[0033] One embodiment is a therapeutic combination of the invention, wherein the therapeutic molecule is a therapeutic molecule of the previous embodiment, the second ligand A2 is a monoclonal antibody in accordance with the invention or at least one binding fragment or domain thereof, a=1, d=0, b=0, h=0, e=0, L7 is a scaffold in accordance with the invention and f=1 or L7 is absent and f=0, L8 is a linker or cleavable linker in accordance with the invention and i=1, c=2-4, when f=1 g=2-4, when f=0 g=0, and saponin C is a saponin in accordance with the invention, preferably SO1861 and / or QS-21, and wherein ligand A1 and ligand A2 are the same or different.

[0034] Certain aspects of the present invention relate to therapeutic combinations, comprising: (a) a first pharmaceutical composition comprising a therapeutic molecule having the chemical structure of Compound I in accordance with the present invention, the first pharmaceutical composition optionally further comprising a pharmaceutically acceptable excipient; and (b) a second pharmaceutical composition comprising a second therapeutic molecule having the chemical structure of Compound II in accordance with the present invention, the second pharmaceutical composition optionally further comprising a pharmaceutically acceptable excipient.

[0035] One aspect of the present invention relates to a first pharmaceutical composition of the present invention for use as a medicament.

[0036] One aspect of the present invention relates to a therapeutic combination for use in treating or preventing cancer in a human subject, the therapeutic combination comprising (a) a first pharmaceutical composition of the present invention and (b) a second pharmaceutical composition of the present invention, wherein ligand A1 or B1 and ligand A2 or B2 are capable of binding to a tumor cell epitope, preferably a tumor cell-specific epitope, on a tumor cell surface molecule, preferably on a tumor cell-specific surface molecule, with the proviso that the tumor cell epitope or tumor cell-specific epitope to which ligand A1 or B1 can bind is the same as or different from the tumor cell epitope or tumor cell-specific epitope to which ligand A2 or B2 can bind.

[0037] One aspect of the present invention relates to a first pharmaceutical composition of the present invention, further comprising a second therapeutic agent molecule of the present invention.

[0038] One aspect of the present invention relates to a first pharmaceutical composition of the present invention, further comprising a second therapeutic drug molecule of the present invention, for use as a medicament.

[0039] One aspect of the present invention relates to a first pharmaceutical composition of the present invention, further comprising a second therapeutic molecule of the present invention, for use in treating or preventing cancer in a human subject.

[0040] Certain aspects of the invention relate to any of the following ADCs and AOCs and semi-finished conjugates thereof, including conjugates comprising a covalently linked saponin of the invention, and / or including conjugates comprising a payload or effector moiety linked to, for example, an antibody of the invention, and each optionally further comprising either at least one effector molecule of the invention or optionally further comprising at least one saponin of the invention, or both:

[0041] Anti-EGFR antibody-saponin; anti-EGFR antibodies - triterpenoid saponins and / or bisdesmosidic triterpene saponins belonging to the 12,13-dehydrooleanane type, having an aldehyde functional group at position C-23 and optionally containing a glucuronic acid functional group on the carbohydrate substituent at the C-3 beta-OH group of the saponin; Anti-EGFR antibody-SO1861; Anti-EGFR antibody-GE1741; Anti-EGFR antibody-SA1641; Anti-EGFR antibody-Quil-A; Anti-EGFR antibody-QS-21; Anti-EGFR antibodies - saponins in the water-soluble saponin fraction of Quillaja saponaria;

[0042] Cetuximab-saponin; Cetuximab - triterpenoid saponins and / or bisdesmosidic triterpene saponins belonging to the 12,13-dehydrooleanane type having an aldehyde functional group at position C-23 and optionally containing a glucuronic acid functional group on the carbohydrate substituent at the C-3 beta-OH group of the saponin; Cetuximab-SO1861; Cetuximab-GE1741; Cetuximab-SA1641; Cetuximab-Quil-A; Cetuximab-QS-21; Cetuximab - saponin in the water-soluble saponin fraction of Quillaja saponaria;

[0043] Anti-HER2 antibody-saponin; Anti-HER2 antibodies - triterpenoid saponins and / or bisdesmosidic triterpene saponins belonging to the 12,13-dehydrooleanane type having an aldehyde functional group at position C-23 and optionally containing a glucuronic acid functional group on the carbohydrate substituent at the C-3 beta-OH group of the saponin; Anti-HER2 antibody-SO1861; Anti-HER2 antibody-GE1741; Anti-HER2 antibody-SA1641; Anti-HER2 antibody-Quil-A; Anti-HER2 antibody-QS-21; Anti-HER2 antibody - saponin in the water-soluble saponin fraction of Quillaja saponaria;

[0044] Trastuzumab-saponin; trastuzumab - triterpenoid saponins and / or bisdesmosidic triterpene saponins belonging to the type of 12,13-dehydrooleananes having an aldehyde functional group at position C-23 and optionally containing a glucuronic acid functional group on the carbohydrate substituent at the C-3 beta-OH group of the saponin; trastuzumab-SO1861; trastuzumab-GE1741; trastuzumab-SA1641; trastuzumab-Quil-A; trastuzumab-QS-21; Trastuzumab - saponin in the water-soluble saponin fraction of Quillaja saponaria;

[0045] anti-CD71 antibody-saponin; anti-CD71 antibodies - triterpenoid saponins and / or bisdesmosidic triterpene saponins belonging to the 12,13-dehydrooleanane type having an aldehyde functional group at position C-23 and optionally containing a glucuronic acid functional group on the carbohydrate substituent at the C-3 beta-OH group of the saponin; Anti-CD71 antibody-SO1861; Anti-CD71 antibody-GE1741; Anti-CD71 antibody-SA1641; Anti-CD71 antibody-Quil-A; Anti-CD71 antibody-QS-21; Anti-CD71 antibody - saponin in the water-soluble saponin fraction of Quillaja saponaria;

[0046] OKT-9-saponin; OKT-9-triterpenoid saponins and / or bisdesmosidic triterpene saponins belonging to the type of 12,13-dehydrooleananes having an aldehyde function at position C-23 and optionally containing a glucuronic acid function on the carbohydrate substituent at the C-3 beta-OH group of the saponin; OKT-9-SO1861; OKT-9-GE1741; OKT-9-SA1641; OKT-9-Quil-A; OKT-9-QS-21; OKT-9 - saponin in the water-soluble saponin fraction of Quillaja saponaria;

[0047] anti-EGFR antibody-oligonucleotide; anti-EGFR antibody-antisense oligonucleotide; Anti-EGFR antibody-siRNA; anti-EGFR antibody-antisense BNA; anti-EGFR antibody-antisense BNA (HSP27); Anti-EGFR antibodies-proteinaceous toxins; anti-EGFR antibody-ribosome-inactivating protein; Anti-EGFR antibody-Giantin;Anti-EGFR antibody-Saporin;

[0048] Cetuximab-oligonucleotide; Cetuximab-antisense oligonucleotides; Cetuximab-siRNA; Cetuximab-antisense BNA; Cetuximab-antisense BNA (HSP27); Cetuximab - protein toxin; Cetuximab-ribosome-inactivating protein; Cetuximab-Giantin; Cetuximab-saporin;

[0049] anti-HER2 antibody-oligonucleotide; anti-HER2 antibody-antisense oligonucleotide; Anti-HER2 antibody-siRNA; anti-HER2 antibody-antisense BNA; anti-HER2 antibody-antisense BNA (HSP27); Anti-HER2 antibody-protein toxin; anti-HER2 antibody-ribosome-inactivating protein; Anti-HER2 antibody-Giantin; Anti-HER2 antibody-Saporin;

[0050] trastuzumab-oligonucleotide; trastuzumab-antisense oligonucleotide; trastuzumab-siRNA; trastuzumab-antisense BNA; trastuzumab-antisense BNA(HSP27); Trastuzumab - protein toxin; trastuzumab-ribosome-inactivating protein; trastuzumab-giantine; trastuzumab-saporin;

[0051] anti-CD71 antibody-oligonucleotide; anti-CD71 antibody-antisense oligonucleotide; Anti-CD71 antibody-siRNA; anti-CD71 antibody-antisense BNA; anti-CD71 antibody-antisense BNA (HSP27); anti-CD71 antibody-proteinaceous toxin; anti-CD71 antibody-ribosome-inactivating protein; anti-CD71 antibody-Giantin; anti-CD71 antibody-Saporin;

[0052] OKT-9-oligonucleotide; OKT-9-antisense oligonucleotide; OKT-9-siRNA; OKT-9-antisense BNA; OKT-9-antisense BNA(HSP27); OKT-9 - protein toxin; OKT-9 - ribosome-inactivating protein; OKT-9-jianthine; OKT-9-saporin;

[0053] Anti-EGFR antibody (-oligonucleotide) (-saponin), where the oligonucleotide is any one or more of an antisense oligonucleotide, siRNA, antisense BNA, and antisense BNA (HSP27), the saponin is any one or more of a triterpenoid saponin and / or a bisdesmosidic triterpene saponin belonging to the 12,13-dehydrooleanane type having an aldehyde functional group at position C-23 and optionally containing a glucuronic acid group on the carbohydrate substituent at the C-3 beta-OH group of the saponin, SO1861, GE1741, SA1641, Quil-A, QS-21, and saponins in the water-soluble saponin fraction of Quillaja saponaria, and the anti-EGFR antibody is preferably cetuximab;

[0054] anti-EGFR antibody (-proteinaceous toxin) (-saponin), where the proteinaceous toxin is any one or more of ribosome-inactivating proteins, dianthins, and saporins, and the saponins are any one or more of triterpenoid saponins and / or bisdesmosidic triterpene saponins belonging to the 12,13-dehydrooleanane type having an aldehyde functional group at position C-23 and optionally containing a glucuronic acid group on the carbohydrate substituent at the C-3 beta-OH group of the saponin, SO1861, GE1741, SA1641, Quil-A, QS-21, and saponins in the water-soluble saponin fraction of Quillaja saponaria, and the anti-EGFR antibody is preferably cetuximab;

[0055] Anti-HER2 antibody (-oligonucleotide) (-saponin), wherein the oligonucleotide is any one or more of an antisense oligonucleotide, siRNA, antisense BNA, and antisense BNA (HSP27), the saponin is any one or more of a triterpenoid saponin and / or a bisdesmosidic triterpene saponin belonging to the 12,13-dehydrooleanane type having an aldehyde functional group at position C-23 and optionally containing a glucuronic acid group on the carbohydrate substituent at the C-3 beta-OH group of the saponin, SO1861, GE1741, SA1641, Quil-A, QS-21, and saponins in the water-soluble saponin fraction of Quillaja saponaria, and the anti-HER2 antibody is preferably trastuzumab;

[0056] anti-HER2 antibody (-proteinaceous toxin) (-saponin), where the proteinaceous toxin is any one or more of ribosome-inactivating proteins, dianthins, and saporins, and the saponins are any one or more of triterpenoid saponins and / or bisdesmosidic triterpene saponins belonging to the 12,13-dehydrooleanane type having an aldehyde functional group at position C-23 and optionally containing a glucuronic acid group on the carbohydrate substituent at the C-3 beta-OH group of the saponin, SO1861, GE1741, SA1641, Quil-A, QS-21, and saponins in the water-soluble saponin fraction of Quillaja saponaria, and the anti-HER2 antibody is preferably trastuzumab;

[0057] Anti-CD71 antibody (-oligonucleotide) (-saponin). The oligonucleotide is any one or more of an antisense oligonucleotide, siRNA, antisense BNA, and antisense BNA (HSP27), the saponin is any one or more of a triterpenoid saponin and / or a bisdesmosidic triterpene saponin belonging to the 12,13-dehydrooleanane type having an aldehyde functional group at position C-23 and optionally containing a glucuronic acid group on the carbohydrate substituent at the C-3 beta-OH group of the saponin, SO1861, GE1741, SA1641, Quil-A, QS-21, and a saponin in the water-soluble saponin fraction of Quillaja saponaria, and the anti-CD71 antibody is preferably OKT-9; and an anti-CD71 antibody (-proteinaceous toxin) (-saponin). The proteinaceous toxin is any one or more of ribosome-inactivating proteins, dianthins, and saporins, the saponins are triterpenoid saponins and / or bisdesmosidic triterpene saponins belonging to the 12,13-dehydrooleanane type having an aldehyde functional group at position C-23 and optionally containing a glucuronic acid group on the carbohydrate substituent at the C-3 beta-OH group of the saponin, SO1861, GE1741, SA1641, Quil-A, QS-21, and saponins in the water-soluble saponin fraction of Quillaja saponaria, and the anti-CD71 antibody is preferably OKT-9.

[0058] One embodiment is a first proteinaceous molecule of the invention, a semi-finished conjugate of the invention, or a conjugate of the invention, wherein the first binding site is selected from cetuximab, trastuzumab, OKT-9, and / or the effector molecule is selected from dianthin, saporin, and antisense BNA (HSP27), and / or the saponin is selected from SO1861, GE1741, SA1641, Quil-A, QS-21, and saponins in the water-soluble saponin fraction of Quillaja saponaria.

[0059] One embodiment is a conjugate according to the invention, wherein the first proteinaceous molecule is selected from cetuximab, trastuzumab, OKT-9, and / or the effector molecule is selected from dianthin, saporin, and antisense BNA (HSP27), and / or the saponin is selected from SO1861, GE1741, SA1641, Quil-A, QS-21, and a saponin in the water-soluble saponin fraction of Quillaja saponaria.

[0060] One aspect of the invention relates to an ADC or AOC or semi-finished ADC conjugate or semi-finished AOC conjugate of structure C, comprising an antibody-saponin conjugate of the invention, comprising at least one effector molecule of the invention, and / or comprising at least one saponin of the invention: A(-S)b(-E)c

[0061] Structure C During the ceremony, A is the first binding site; S is saponin; E is an effector molecule; b=0 to 64, preferably 0, 1, 2, 3, 4, 8, 16, 32, 64, or any integer or fraction therebetween; c=0-8, preferably 0, 1, 2, 3, 4, 6, 8, or any integer or fraction therebetween, and S is coupled to A and / or E, and E is coupled to A and / or S, preferably S is coupled to A and E is coupled to A.

[0062] One embodiment is structure C of the invention, wherein A is an anti-EGFR antibody, e.g., cetuximab, an anti-HER2 antibody, e.g., trastuzumab, an anti-CD71 antibody, e.g., OKT-9, and / or S is a saponin, triterpenoid saponin, and / or bisdesmosidic triterpene saponin belonging to the type of 12,13-dehydrooleanane having an aldehyde functional group at position C-23 and optionally containing a glucuronic acid functional group on the carbohydrate substituent at the C-3 beta-OH group of the saponin, such as SO1861, GE1741, SA1641, Quil-A, QS-21, and Quillaja. and / or E is any one or more of the saponins in the water-soluble saponin fraction of S. saponaria, and / or E is any one or more of an oligonucleotide, an antisense oligonucleotide, an siRNA, an antisense BNA, and an antisense BNA (HSP27), and / or any one or more of a proteinaceous toxin, a ribosome-inactivating protein, a dianthin, and a saporin.

[0063] An embodiment is Structure C of the invention, a conjugate of the invention, or a semi-finished conjugate of the invention, or a first proteinaceous molecule of the invention, wherein the saponin, if present, and / or the effector molecule, if present, are covalently coupled via at least one linker, such as a cleavable linker, and / or at least one oligomeric or polymeric backbone, such as N-ε-maleimidocaproic acid hydrazide (EMCH), succinimidyl 3-(2-pyridyldithio)propionate or 3-(2-pyridyldithio)propionic acid N-hydroxysuccinimide ester (SPDP), and 1-[bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium 3-oxide hexafluorophosphate (HATU)-based linkers, and for example, a dendron-based backbone, such as a G4 dendron, or a trifunctional linker, such as the trifunctional linker of Scheme II. and / or at least one lysine and / or cysteine ​​side chain of the first binding site of the antibody, preferably a monoclonal antibody or fragment or domain thereof, is involved in a covalent bond to the saponin and / or effector molecule and / or linker and / or cleavable linker and / or scaffold. Preferably, the saponin and / or effector molecule is covalently linked to the first binding site of the antibody, preferably a monoclonal antibody, the covalent linkage comprising or consisting of an amide bond, hydrazone bond or disulfide bond.

[0064] One aspect of the present invention relates to the use of any of the previously mentioned conjugates of the invention, or any of the semi-finished conjugates of the invention, or the antibody-saponin conjugates of the invention, as a medicament.

[0065] One aspect of the present invention relates to the use of either a conjugate of the present invention or a semi-finished conjugate of the present invention or an antibody-saponin conjugate of the present invention for use in the treatment or prevention of cancer or an autoimmune disease.

[0066] Figures 25 and 26 show examples of ADCs of the invention having covalently coupled saponin(s) and OACs of the invention having covalently coupled saponin(s).

[0067] It will be understood that any and all of a, b, c, d, e, f, g, h, I, j, k, m, n, p, q, r, s, t, u, v, w, and / or x have values ​​according to each individual embodiment and aspect of the present invention for any and all of the previously mentioned embodiments and aspects according to the present invention. In addition, as will be readily understood by those skilled in the art, the (trifunctional) linkers L1, L2, L4, L5, L6, L8, L9, and / or L10, when present in a molecule or conjugate or moiety of the present invention, are the (trifunctional) linkers indicated for each and any of the previously mentioned aspects and embodiments of the present invention. Also, as will be readily understood by those skilled in the art, the oligomeric or polymeric backbone L3 and / or L7, when present in a molecule or conjugate or moiety of the present invention, are the oligomeric or polymeric backbone indicated for each and any of the previously mentioned aspects and embodiments of the present invention. Furthermore, the first ligand A1 and first effector moiety B1, if present, and the second ligand A2 and second effector moiety B2, if present, and the first effector moiety A1 and first ligand B1, if present, and the second effector moiety A2 and second ligand moiety B2, if present, are ligands and effector moieties selected and indicated as disclosed for the first, second, third, fourth, fifth and sixth series of embodiments and aspects of the invention as outlined herein above, as well as all further embodiments and aspects of the invention. Saponin C is any one or more of the saponins referenced and listed in any of the previously mentioned aspects and embodiments of the invention, in particular one or more saponins selected from Scheme I and / or Table A1.

[0068] definition The term "linker" has its usual scientific meaning and herein refers to a linear stretch or chemical moiety of amino acid residues conjugated via a peptide bond, which attaches a molecule or atom to another molecule, such as a ligand or an effector molecule or a scaffold. Typically, a linker comprises a chain of atoms linked by a chemical bond. Any linker molecule or linker technology known in the art can be used in the present disclosure. Where indicated, a linker is a linker for covalently attaching molecules via a chemical group on such molecule suitable for forming a covalent bond or covalent linkage with the linker. The linker can be a non-cleavable linker. For example, the linker is stable under physiological conditions. The linker can be a cleavable linker, for example, a linker that is cleavable in the presence of an enzyme, at a specific pH range or value, or under physiological conditions, such as the intracellular conditions of endosomes, such as lysosomes and late endosomes, of mammalian cells, such as human cells. Exemplary linkers that may be used in the context of the present disclosure include, but are not limited to, N-ε-maleimidocaproic acid hydrazide (EMCH), succinimidyl 3-(2-pyridyldithio)propionate, or 3-(2-pyridyldithio)propionic acid N-hydroxysuccinimide ester (SPDP), and 1-[bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium 3-oxide hexafluorophosphate (HATU).

[0069] The term "trifunctional linker" has its usual scientific meaning and refers herein to a linker that attaches three molecules via their respective chemical groups. Those skilled in the art can design such trifunctional linkers based on this disclosure and common general knowledge. Such trifunctional linkers may exhibit, for example, maleimide groups that can be used for conjugation to targeting ligands that exhibit thiol groups for thiol-ene reactions. Additionally, trifunctional linkers may exhibit dibenzocyclooctyne (DBCO) groups for so-called strain-promoted alkyne-azide cycloaddition (SPAAC, click chemistry) with azide-bearing saponins. Finally, trifunctional linkers may acquire a third functional group, such as a trans-cyclooctene (TCO) group, to undergo so-called inverse electron-demand Diels-Alder (IEDDA) reactions with tetrazine (Tz)-bearing effector molecules. Those skilled in the art will understand that the three chemical groups of a trifunctional linker can all be the same or different, or the linker can contain two of the same chemical groups for linking molecules to the trifunctional linker. The bond formed between the trifunctional linkers can be covalent or non-covalent, with covalent bonds being preferred. The bond formed between the trifunctional linker and one, two, or three binding molecules via each chemical group can be a cleavable (labile) bond, cleavable under acidic conditions in cells, such as the endosomes and lysosomes of mammalian cells, such as human cells, or a non-cleavable bond. It should be understood that a trifunctional linker can contain one or two chemical groups for forming a covalent bond, and two or one additional chemical group(s) for forming a non-covalent bond, respectively. It will be appreciated that a trifunctional linker may include one or two chemical groups for forming a cleavable bond, and an additional two or one chemical group(s), respectively, for forming a non-cleavable bond.

[0070] The term "cleavable," as used, for example, in the term "cleavable linker" or "cleavable bond," has its usual scientific meaning, and herein refers to cleavage under acidic, reductive, enzymatic, or photoinduced conditions. For example, a cleavable linker may be cleaved under acidic conditions. Preferably, the cleavable linker is cleaved in vivo under acidic conditions present in endosomes and / or lysosomes of mammalian cells, preferably human cells, preferably at a pH of 4.0 to 6.5, more preferably at a pH of 5.5 or less. As another example, the cleavable linker may be cleaved by an enzyme, for example, a cathepsin. Furthermore, an example of a covalent bond cleavable under reductive conditions is a disulfide bond.

[0071] The terms "oligomer" and "polymer" in the context of oligomeric or polymeric backbones have their customary scientific meanings. Here, a polymer refers to a substance having a molecular structure built up primarily or entirely from a large number of equal or similar units linked together; here, an oligomer refers to a polymer whose molecules consist of a relatively small number of repeating units. For example, a structure containing 5-10 or fewer equal or similar units may be referred to as an oligomeric structure, a structure containing 10-50 or more monomeric units may be referred to as a polymeric structure, and a structure of 10 monomeric units may be referred to as either oligomeric or polymeric.

[0072] The term "binding site" has its usual scientific meaning and refers here to a region or epitope on one molecule, such as a protein, DNA, or RNA, to which another molecule can bind.

[0073] The term "scaffold" has its usual scientific meaning and herein refers to an oligomeric or polymeric template or carrier or base (base molecule or base structure) to which one or more molecules, e.g., ligand molecules, effector molecules, can be covalently attached either directly or via a linker, such as a cleavable linker. The scaffold may have a structurally ordered formation, e.g., a polymer, oligomer, dendrimer, dendronized polymer, or dendronized oligomer, or may comprise an assembled polymeric structure, e.g., a hydrogel, microgel, nanogel, stabilized polymeric micelle, or liposome, but excludes structures consisting of non-covalent aggregates of monomers, such as cholesterol / phospholipid mixtures. The backbone may comprise polymeric or oligomeric structures, such as poly- or oligo(amines), e.g., polyethyleneimine and poly(amidoamine); or polyethylene glycol, poly- or oligo(esters), e.g., poly(lactides), poly(lactams), polylactide-co-glycolide copolymers; or poly(dextrins), poly- or oligosaccharides, e.g., cyclodextrin or polydextrose; or natural and / or artificial poly- or oligoamino acids, e.g., polylysine, or peptides or proteins, DNA oligo- or polymers, stabilized RNA polymers, or PNA (peptide nucleic acid) polymers. Preferably, the polymeric or oligomeric structures are biocompatible, meaning that they do not exhibit substantial acute or chronic toxicity in organisms and can either be excreted intact or can be completely degraded by the body's metabolism into excretable and / or physiological compounds.

[0074] The term "ligand" has its usual scientific meaning and refers herein to any molecule or molecules that can selectively bind to a target cell surface molecule or target cell surface receptor expressed on a target cell, e.g., a target cancer cell or a target autoimmune cell. A ligand can bind to an epitope contained by a receptor or other antigen on a target cell. Preferably, the cell-binding ligand is an antibody.

[0075] As used herein, the term "antibody" is used in the broadest sense and can refer to an immunoglobulin (Ig), defined as a protein belonging to the classes IgG, IgM, IgE, IgA, or IgD (or any subclass thereof), or to a functional binding fragment or binding domain of an immunoglobulin. In the context of the present invention, a "binding fragment" or "binding domain" of an immunoglobulin is defined as an antigen-binding fragment or domain of a parent immunoglobulin or other derivative, which essentially retains the antigen-binding activity of such parent immunoglobulin. Functional fragments and functional domains are antibodies in the sense of the present invention, even if their affinity for antigen is lower than that of the parent immunoglobulin. "Functional fragments and domains" according to the present invention include, but are not limited to, F(ab')2 fragments, Fab' fragments, Fab fragments, scFv, dsFv, single-domain antibodies (sdAbs), monovalent IgG, scFv-Fc, reduced IgG (rIgG), minibodies, diabodies, triabodies, tetrabodies, Fc fusion proteins, nanobodies, variable V domains such as VHH and Vh, and other types of antigen-recognizing immunoglobulin fragments and domains. Fragments and domains can be engineered to minimize or completely eliminate intermolecular disulfide interactions that occur between CH1 and CL domains. Due to their smaller size, functional fragments and domains offer the advantage of greater tumor penetration. In addition, functional fragments or domains may be more uniformly distributed in tumor masses compared to whole immunoglobulins.

[0076] The antibodies (immunoglobulins) of the present invention may be bifunctional or multifunctional. For example, a bifunctional antibody has one arm with specificity for one receptor or antigen, and the other arm recognizes a different receptor or antigen. Alternatively, each arm of a bifunctional antibody may have specificity for a different epitope of the same receptor or antigen on a target cell.

[0077] The antibodies (immunoglobulins) of the present invention can be, but are not limited to, polyclonal antibodies, monoclonal antibodies, human antibodies, humanized antibodies, chimeric antibodies, resurfaced antibodies, anti-idiotypic antibodies, mouse antibodies, rat antibodies, rat / mouse hybrid antibodies, llama antibodies, heavy chain-only llama antibodies, heavy chain-only antibodies, and veterinary antibodies. Preferably, the antibodies (immunoglobulins) of the present invention are monoclonal antibodies. Resurfaced, chimeric, humanized, and fully human antibodies are also more preferred because they are less likely to cause immunogenicity in humans. The antibodies of the ADCs of the present invention preferably specifically bind to antigens expressed on the surface of cancer cells, autoimmune cells, diseased cells, and abnormal cells while leaving any healthy cells essentially unchanged (e.g., by not binding to such normal cells or by binding to such healthy cells with a lesser degree of number and / or affinity).

[0078] Specific antibodies that can be used in the ADCs of the invention include anti-HER2 monoclonal antibodies, such as trastuzumab and pertuzumab, anti-CD20 monoclonal antibodies, such as rituximab, ofatumumab, tositumomab, and ibritumomab, anti-CA125 monoclonal antibodies, such as oregovomab, anti-EpCAM (17-1A) monoclonal antibodies, such as edrecolomab, anti-EGFR monoclonal antibodies, such as cetuximab, panitumumab, and nimotuzumab, anti-CD30 monoclonal antibodies, such as brentuximab, anti-CD33 monoclonal antibodies, such as gemtuzumab and huMy9-6, anti-vascular integrin alpha-v beta-3 monoclonal antibodies, such as etaracizumab, and anti-CD52 monoclonal antibodies, such as alemtuzumab. anti-CD22 monoclonal antibodies, such as epratuzumab, anti-CEA monoclonal antibodies, such as labetuzumab, anti-CD44v6 monoclonal antibodies, such as bivatuzumab, anti-FAP monoclonal antibodies, such as sibrotuzumab, anti-CD19 monoclonal antibodies, such as huB4, anti-CanAg monoclonal antibodies, such as huC242, anti-CD56 monoclonal antibodies, such as huN901, anti-CD38 monoclonal antibodies, such as daratumumab, anti-CA6 monoclonal antibodies, such as DS6, anti-IGF-IR monoclonal antibodies, such as cixutumumab and 3B7, anti-integrin monoclonal antibodies, such as CNTO95, and anti-syndecan-1 monoclonal antibodies, such as B-B4.

[0079] Any molecule other than an antibody that binds to a cell receptor or antigen on a target cell can also be used as the cell-binding ligand of the ligand-drug conjugate of the present invention. The ligand is provided in accordance with the present invention as a covalently attached saponin. These ligands include, but are not limited to, proteins, polypeptides, peptides, and small molecules. Examples of these non-antibody ligands include interferons (e.g., IFN-α, IFN-β, and IFN-γ), transferrin, lectins, epidermal growth factor (EGF) and EGF-like domains, gastrin-releasing peptide (GRP), platelet-derived growth factor (TGF), transforming growth factor (TGF), vaccinia growth factor (VGF), insulin and insulin-like growth factors (IGFs, e.g., IGF-1 and IGF-2), other suitable hormones, such as thyrotropin-releasing hormone (TRH), melanocyte-stimulating hormone (MSH), and steroids. Hormones (e.g., estrogen and androgen), somatostatin, lymphokines (e.g., IL-2, IL-3, IL-4, and IL-6), colony-stimulating factors (CSFs, e.g., G-CSF, M-CSF, and GM-CSF), bombesin, gastrin, Arg-Gly-Asp or RGD, aptamers (e.g., AS-1411, GBI-10, RNA aptamers against HIV glycoproteins), small molecules (e.g., folic acid, anisamidophenylboronic acid), vitamins (e.g., vitamin D), carbohydrates (e.g., hyaluronic acid, galactose).

[0080] An "effector molecule" or "effector moiety" or "payload" has its usual scientific meaning and, in the context of the present invention, is any substance that affects cellular metabolism by interacting with an effector molecule target within the cell. This effector molecule target is any molecule or structure within the cell that excludes the lumen of compartments and vesicles of the endocytic and recycling pathways, but includes the membranes of these compartments and vesicles. Said structures within the cell therefore include the nucleus, mitochondria, chloroplasts, endoplasmic reticulum, Golgi apparatus, other transport vesicles, the interior of the plasma membrane, and the cytosol.

[0081] The effector molecule or moiety is a drug substance, such as a toxin, e.g., a proteinaceous toxin, a drug, a polypeptide, or a polynucleotide. A drug substance in the present invention is an effector molecule or moiety used to achieve a beneficial outcome in an organism, preferably a vertebrate, more preferably a mammal, such as a non-human subject or a human / human subject. Benefits include diagnosis, prognosis, treatment, cure, and prevention (prophylaxis) of a disease and / or condition and / or health problem. A drug substance may also lead to unwanted and possibly even harmful side effects (e.g., adverse events observed during clinical trials). In this case, the pros and cons must be weighed to determine whether the drug substance is suitable in a particular case. If the effects of a drug substance in a cell are overwhelmingly beneficial to the organism as a whole, the cell is called a target cell. If the effects in a cell are overwhelmingly harmful to the organism as a whole, the cell is called an off-target cell. In artificial systems such as cell culture and bioreactors, target and off-target cells depend on the purpose and are defined by the user. Examples of effector molecules and moieties are drugs, toxins, polypeptides (eg, enzymes), polynucleotides (including polypeptides and polynucleotides that contain unnatural amino acids or nucleic acids), and any combination thereof.

[0082] Drug effector molecules or effector moieties may include, but are not limited to, anti-cancer agents, anti-inflammatory agents, and anti-infective (e.g., anti-fungal, anti-bacterial, anti-parasitic, anti-viral) agents. Preferably, the drug molecules of the present invention are anti-cancer or anti-autoimmune agents. Suitable anti-cancer agents include, but are not limited to, alkylating agents, metabolic inhibitors, spindle poison plant alkaloids, cytotoxic / anti-tumor antibiotics, topoisomerase inhibitors, photosensitizers, and kinase inhibitors. The definition of "anti-cancer agent" also includes, for example, (i) antihormonal agents that act to control or inhibit hormone action on tumors, such as antiestrogens and selective estrogen receptor modulators; (ii) aromatase inhibitors that inhibit the enzyme aromatase, which controls estrogen production in the adrenal glands; (iii) antiandrogens; (iv) protein kinase inhibitors; (v) lipid kinase inhibitors; (vi) antisense oligonucleotides, particularly those that inhibit the expression of genes in signal transduction pathways involved in abnormal cell proliferation; (vii) ribozymes, such as VEGF expression inhibitors and HER2 expression inhibitors; (viii) vaccines, such as gene therapy vaccines; topoisomerase 1 inhibitors; (ix) angiogenesis inhibitors; and pharmaceutically acceptable salts, acids, solvates, and derivatives of any of the above.

[0083] Toxin effector molecules or moieties can include, but are not limited to, proteinaceous toxins (e.g., bacterial and plant-derived toxins), toxins that target tubulin filaments, toxins that target DNA, and toxins that target RNA. Examples of proteinaceous toxins include saporin, dianthin, ricin, modeccin, abrin, volkensin, viscumin, Shiga toxin, Shiga-like toxins, Pseudomonas exotoxin (PE, also known as exotoxin A), diphtheria toxin (DT), and cholera toxin. Examples of tubulin filament-targeting toxins include maytansinoids (e.g., DM1 and DM4), auristatins (e.g., monomethylauristatin E (MMAE) and monomethylauristatin F (MMAF)), toxoids, tubulysins, cryptophycins, and rhizoxins. Examples of DNA-targeted toxins are calicheamicin: N-acetyl-γ-calicheamicin, CC-1065 analogs, duocarmycin, doxorubicin, methotrexate, benzodiazepines, camptothecin analogs, and anthracyclines. Examples of DNA-targeted toxins are amanitin, splicehostatin, and Thailanstatin. The toxins used in the present invention are defined as drug substances that can kill or inactivate cells. Preferably, targeted toxins are toxins that are only, or at least predominantly, toxic to target cells, but not to off-target cells. The net effect of the targeted toxin is preferably beneficial to the organism as a whole.

[0084] An effector molecule or moiety that is a polypeptide can be, for example, a polypeptide that restores a lost function, such as, for example, an enzyme replacement or a gene regulatory function, or a toxin. Examples of polypeptides as effector molecules include, for example, Cas9; toxins (e.g., saporin, dianthin, gelonin, (de)bouganin, agrostin, ricin (toxin A chain); pokeweed antiviral protein, apoptin, diphtheria toxin, Pseudomonas exotoxin), metabolic enzymes (e.g., argininosuccinate lyase, argininosuccinate synthetase), enzymes of the coagulation cascade, repair enzymes; enzymes of cell signaling; cell cycle regulators; gene regulators (transcription factors, e.g., NF-κB, or gene repressors, e.g., methionine repressor).

[0085] An effector molecule or effector moiety that is a polynucleotide can be, for example, a polynucleotide containing coding information, such as a gene or open reading frame encoding a protein. It can also contain regulatory information, such as a promoter or regulatory element binding region, or a sequence encoding a microRNA. Such polynucleotides can include natural and artificial nucleic acids. Artificial nucleic acids include, for example, peptide nucleic acids (PNAs), morpholinos and locked nucleic acids (LNAs), as well as glycol nucleic acids (GNAs) and threose nucleic acids (TNAs). Each of these is distinguished from naturally occurring DNA or RNA by modifications to the backbone of the molecule. Examples of nucleotides as effector molecules include, but are not limited to, DNA: single-stranded DNA (e.g., adenine phosphoribosyltransferase DNA); linear double-stranded DNA (e.g., coagulation factor IX gene); circular double-stranded DNA (e.g., plasmid); RNA: mRNA (e.g., TAL effector molecule nuclease), tRNA, rRNA, siRNA, miRNA, antisense RNA; antisense oligonucleotides (ASO, AON, e.g., PNA, PMO, LNA, and BNA).

[0086] For example, the term "proteinaceous," as used in "proteinaceous molecule" and "proteinaceous toxin," refers to molecules and toxins comprising at least one string of amino acid residues that can be obtained as an expression product from a single mRNA. Such molecules or toxins can further comprise any post-translational modifications, carbohydrates, e.g., N- or O-linked carbohydrates, disulfide bonds, phosphorylation, sulfation, etc., resulting from any post-translational modification, and / or can further comprise any other modifications, e.g., resulting from chemical modifications (e.g., resulting from the attachment of an effector moiety, saponin, scaffold, ligand, etc., either directly, e.g., to an amino acid side chain, or via at least one linker that is (covalently) bound to the molecule and chemically bound to the proteinaceous molecule to chemically modify the proteinaceous molecule). The term "proteinaceous" also encompasses and includes aggregates of such molecules, e.g., homodimers, heterotrimers, heterohexamers, or complex assemblies, e.g., ribosomes.

[0087] For example, in the context of "specific binding" and "a receptor or molecular target specifically present or expressed on the surface of tumor cells," and the like, the terms "specific" and "specifically" have their ordinary scientific meaning known in the art. Here, for example, it refers to a binding interaction of a first molecule with a second molecule that occurs with a relatively higher affinity relative to any putative binding of the first molecule to an additional molecule different from the second molecule, or, when considering a number of receptors or molecular targets, the expression or higher degree of expression of a cell surface receptor or molecular target on the surface of a first type of cell, such as a tumor cell, autoimmune cell, diseased cell, or abnormal cell, relative to the degree of expression of the same receptor or molecular target on a second type of cell, such as a healthy cell. Expression in a second type of cell may be completely absent or very low relative to any degree of expression on tumor cells. Furthermore, for example, in "specific binding," the term "specific" has its ordinary scientific meaning known in the art, and here refers to a molecule that can interact with another molecule with a higher binding affinity than background interactions between molecules. Similarly, the term "specificity" refers to an interaction, e.g., between two molecules or between a cell and a molecule, that has a higher binding affinity than background interactions between molecules. Binding molecules, such as immunoglobulins, bind via their binding site, e.g., the immunoglobulin variable region of an immunoglobulin, to a binding site on a molecule, e.g., an epitope, cell surface receptor, etc., with a higher binding affinity than background interactions between molecules. In the context of the present invention, background interactions typically have a K of 10E-4 M or less. D Similarly, a "specific binding domain" is a domain that preferentially binds to a binding site on a molecule, e.g., an epitope, cell surface receptor, etc., with a binding affinity higher than background interactions between molecules. In the context of the present invention, a "background interaction" is typically a K of 10E-4 M or less. D Preferably, the specific binding domain has a K of about 10E-5 M. D It binds with higher affinity than

[0088] The term "binding" is defined as an interaction between molecules that can be distinguished from background interactions.

[0089] As used herein, the term "fragment" refers to an amino acid sequence that is part of a protein domain or that makes up an intact protein domain. Binding fragments according to the invention must have binding specificity for their respective targets, e.g., cell surface receptors on the surface of diseased cells, e.g., tumor cells.

[0090] The term "ADC" or "antibody-drug conjugate" has its usual scientific meaning known to those of skill in the art and, herein, refers to a class of biopharmaceutical drugs designed as targeted therapies, for example, to treat cancer. Unlike chemotherapy, ADCs are intended to target and kill tumor cells while sparing healthy cells. ADCs consist of an antibody linked to a bioactive cytotoxic (anticancer) payload or drug. ADCs combine the targeting capabilities of monoclonal antibodies with the cancer-killing capabilities of cytotoxic drugs. They are designed with the intent of discriminating between healthy cells and diseased tissue, such as tumor cells in tumors.

[0091] The term "Saponinum album" has its ordinary meaning and herein refers to a mixture of saponins produced by Merck KGaA (Darmstadt, Germany) containing saponins from Gypsophila paniculata and Gypsophila arostii, including SA1657 and primarily SA1641.

[0092] The term "Quillaja saponin" has its ordinary meaning and refers herein to the saponin fraction of Quillaja saponaria, and therefore the source of all other QS saponins, containing primarily QS-18 and QS-21.

[0093] "QS-21" or "QS21" has its customary scientific meaning and refers herein to a mixture of QS-21 A-apio (~63%), QS-21 A-xylo (~32%), QS-21 B-apio (~3.3%), and QS-21 B-xylo (~1.7%).

[0094] Similarly, "QS-21A" has its customary scientific meaning and refers herein to a mixture of QS-21 A-apio (-65%) and QS-21 A-xylo (-35%).

[0095] Similarly, "QS-21B" has its customary scientific meaning and refers herein to a mixture of QS-21 B-apio (-65%) and QS-21 B-xylo (-35%).

[0096] The term "Quill-A" refers to a commercially available semi-purified extract from Quillaja saponaria, which contains variable quantities of more than 50 distinct saponins, many of which incorporate the triterpene-trisaccharide substructure Gal-(1→2)-[Xyl-(1→3)]-GlcA- at the C-3 beta-OH group, found in QS-7, QS-17, QS18, and QS-21. The saponins found in Quil-A are listed in Table 2 of van Setten (1995) (Dirk C. van Setten, Gerrit van de Werken, Gijsbert Zomer, and Gideon FA Kersten, Glycosyl Compositions and Structural Characteristics of the Potential Immuno-adjuvant Active Saponins in the Quillaja saponaria Molina Extract Quil A, RAPID COMMUNICATIONS IN MASS SPECTROMETRY, VOL. 9, 660-666 (1995)). Quil-A and Quillaja saponin are also fractions of saponins from Quillaja saponaria, and both contain a large variety of different saponins with largely overlapping contents. Because the two fractions are obtained by different purification procedures, the two fractions differ in their specific composition.

[0097] The terms "QS1861" and "QS1862" refer to QS-7 and QS-7 api. QS1861 has a molecular mass of 1861 daltons, and QS1862 has a molecular mass of 1862 daltons. QS1862 is described in Fleck et al. (2019), Table 1, line 28 (Juliane Deise Fleck, Andresa Heemann Betti, Francini Pereira da Silva, Eduardo Artur Troian, Cristina Olivaro, Fernando Ferreira and Simone Gasparin Verza, Saponins from Quillaja saponaria). and Quillaja brasiliensis: Particular Chemical Characteristics and Biological Activities, Molecules 2019, 24, 171; doi:10.3390 / molecules24010171). The structure described is that of the api variant of QS-7, QS1862. The molecular mass is 1862 daltons because this is the nominal mass including the proton in the glucuronic acid. At neutral pH, the molecule is deprotonated. When measured by mass spectrometry in negative ion mode, the measured mass is 1861 daltons.

[0098] The terms first, second, third, and the like in the specification and claims are used to distinguish between similar elements and not necessarily to describe a sequential or chronological order. The terms are interchangeable in appropriate circumstances. Embodiments of the invention may operate in sequences other than those described or illustrated herein.

[0099] Furthermore, although various embodiments may be referred to as "preferably" or "eg" or "for example" or "specifically," these should not be construed as limiting the scope of the invention, but rather as exemplary ways in which the invention may be implemented.

[0100] The term "comprising" used in the claims should not be construed as being limited to the subsequently listed elements or steps; it does not exclude other elements or steps. It needs to be construed as specifying the presence of a stated feature, integer, step, or component, but does not exclude the presence or addition of one or more other features, integers, steps, or components, or groups thereof. Thus, the scope of the phrase "a pharmaceutical composition comprising A and B" should not be limited to a pharmaceutical composition consisting solely of components A and B; rather, for the purposes of the present invention, the recited components of the pharmaceutical composition are only A and B, and the claim should be construed to encompass equivalents of those components. Similarly, the scope of the phrase "a method comprising steps A and B" should not be limited to a method consisting solely of steps A and B; rather, for the purposes of the present invention, the recited steps of the method are only A and B, and the claim should be construed to encompass equivalents of those steps.

[0101] Additionally, unless the context clearly requires that there be only one of the feature, reference to a feature by the indefinite article "a" or "an" does not exclude the possibility that more than one of the feature may be present, e.g., a component, excipient, saponin, etc. Thus, the indefinite article "a" or "an" normally means "at least one." [Brief explanation of the drawings]

[0102] [Figure 1] Figure 1-1: In vivo HSP27 expression in the A431 xenograph "nude" mouse tumor model treated with 30 mg / kg cetuximab-Cys-(SO1861-L-trifunctional linker-L-HSP27BNA), 25 mg / kg cetuximab-(Lys-L-HSP27BNA)4, or 25 mg / kg cetuximab-(Cys-L-SO1861). [Figure 2]Figure 2-1: Enhanced HSP27 gene silencing in vitro in EGFR-expressing A431 cells by treatment with cetuximab-Cys-(SO1861-L-trifunctional linker-L-HSP27BNA), cetuximab-(Lys-L-HSP27BNA)4, or cetuximab-(Cys-L-SO1861). [Figure 3] Figure 3-1: Legend and axes are the same for Figures A, B, C, and D. A. Cytotoxicity of cetuximab, cetuximab + 10 pM cetuximab-saporin, cetuximab-Cys-(dendron(-L-SO1861)4)3,9, and cetuximab-Cys-(dendron(-L-SO1861)4)3,9 + 10 pM cetuximab-saporin in EGFR-expressing cells (MDA-MB-468). B. Cell-killing activity of trastuzumab, trastuzumab + 50 pM trastuzumab-saporin, trastuzumab-Cys-(dendron(-L-SO1861)4)4, and trastuzumab-Cys-(dendron(-L-SO1861)4)4 + 50 pM trastuzumab-saporin in HER2-expressing cells (SK-BR-3). C. Cell-killing activity of cetuximab, cetuximab + 10 pM cetuximab-saporin, cetuximab-Cys-(dendron(-L-SO1861)4)3,9, and cetuximab-Cys-(dendron(-L-SO1861)4)3,9 + 10 pM cetuximab-saporin in EGFR-expressing cells (HeLa). D. Cytotoxic activity of trastuzumab, trastuzumab + 50 pM trastuzumab-saporin, trastuzumab-Cys-(dendron(-L-SO1861)4)4, and trastuzumab-Cys-(dendron(-L-SO1861)4)4 + 50 pM trastuzumab-saporin in HER2-expressing cells (JIMT-1). [Figure 4]Figure 3-1: Legend and axes are the same for Figures A, B, C, and D. A. Cytotoxicity of cetuximab, cetuximab + 10 pM cetuximab-saporin, cetuximab-Cys-(dendron(-L-SO1861)4)3,9, and cetuximab-Cys-(dendron(-L-SO1861)4)3,9 + 10 pM cetuximab-saporin in EGFR-expressing cells (MDA-MB-468). B. Cell-killing activity of trastuzumab, trastuzumab + 50 pM trastuzumab-saporin, trastuzumab-Cys-(dendron(-L-SO1861)4)4, and trastuzumab-Cys-(dendron(-L-SO1861)4)4 + 50 pM trastuzumab-saporin in HER2-expressing cells (SK-BR-3). C. Cell-killing activity of cetuximab, cetuximab + 10 pM cetuximab-saporin, cetuximab-Cys-(dendron(-L-SO1861)4)3,9, and cetuximab-Cys-(dendron(-L-SO1861)4)3,9 + 10 pM cetuximab-saporin in EGFR-expressing cells (HeLa). D. Cytotoxic activity of trastuzumab, trastuzumab + 50 pM trastuzumab-saporin, trastuzumab-Cys-(dendron(-L-SO1861)4)4, and trastuzumab-Cys-(dendron(-L-SO1861)4)4 + 50 pM trastuzumab-saporin in HER2-expressing cells (JIMT-1). [Figure 5]Figure 3-1: Legend and axes are the same for Figures A, B, C, and D. A. Cytotoxicity of cetuximab, cetuximab + 10 pM cetuximab-saporin, cetuximab-Cys-(dendron(-L-SO1861)4)3,9, and cetuximab-Cys-(dendron(-L-SO1861)4)3,9 + 10 pM cetuximab-saporin in EGFR-expressing cells (MDA-MB-468). B. Cell-killing activity of trastuzumab, trastuzumab + 50 pM trastuzumab-saporin, trastuzumab-Cys-(dendron(-L-SO1861)4)4, and trastuzumab-Cys-(dendron(-L-SO1861)4)4 + 50 pM trastuzumab-saporin in HER2-expressing cells (SK-BR-3). C. Cell-killing activity of cetuximab, cetuximab + 10 pM cetuximab-saporin, cetuximab-Cys-(dendron(-L-SO1861)4)3,9, and cetuximab-Cys-(dendron(-L-SO1861)4)3,9 + 10 pM cetuximab-saporin in EGFR-expressing cells (HeLa). D. Cytotoxic activity of trastuzumab, trastuzumab + 50 pM trastuzumab-saporin, trastuzumab-Cys-(dendron(-L-SO1861)4)4, and trastuzumab-Cys-(dendron(-L-SO1861)4)4 + 50 pM trastuzumab-saporin in HER2-expressing cells (JIMT-1). [Figure 6]Figure 4-1: Legend and axes are the same for Figures A, B, C, and D. A. Cytotoxic activity of cetuximab, cetuximab + 10 pM CD71mab-saporin, cetuximab-Cys-(dendron(-L-SO1861)4)3,9, cetuximab-Cys-(dendron(-L-SO1861)4)3,9 + 10 pM CD71mab-saporin, cetuximab-Lys-(dendron(-L-SO1861)4)4,4, and cetuximab-Lys-(dendron(-L-SO1861)4)4,4 + 10 pM CD71mab-saporin in EGFR++ / CD71+ cells (MDA-MB-468). B. Cytotoxic activity of trastuzumab, trastuzumab + 10 pM CD71mab-saporin, trastuzumab-Cys-(dendron(-L-SO1861)4)4, trastuzumab-Cys-(dendron(-L-SO1861)4)4 + 10 pM CD71mab-saporin, trastuzumab-Lys-(dendron(-L-SO1861)4)4,7, or trastuzumab-Lys-(dendron(-L-SO1861)4)4,7 + 10 pM CD71mab-saporin in HER2++ / CD71+ (SK-BR-3) cell line. C. Cytotoxic activity of cetuximab, cetuximab + 10pM CD71mab-saporin, cetuximab-Cys-(dendron(-L-SO1861)4)3,9, cetuximab-Cys-(dendron-L-SO1861)4)3,9 + 10pM CD71mab-saporin, cetuximab-Lys-(dendron(-L-SO1861)4)4,4, or cetuximab-Lys-(dendron(-L-SO1861)4)4,4 + 10pM CD71mab-saporin in EGFR+ / CD71+ cells (CaSki).D. Cytotoxic activity of trastuzumab, trastuzumab + 10 pM CD71mab-saporin, trastuzumab-Cys-(dendron(-L-SO1861)4)4, trastuzumab-Cys-(dendron(-L-SO1861)4)4 + 10 pM CD71mab-saporin, trastuzumab-Lys-(dendron(-L-SO1861)4)4,7, or trastuzumab-Lys-(dendron(-L-SO1861)4)4,7 + 10 pM CD71mab-saporin in HER2+ / - / CD71+ cells (JIMT-1). [Figure 7]Figure 4-1: Legend and axes are the same for Figures A, B, C, and D. A. Cytotoxic activity of cetuximab, cetuximab + 10 pM CD71mab-saporin, cetuximab-Cys-(dendron(-L-SO1861)4)3,9, cetuximab-Cys-(dendron(-L-SO1861)4)3,9 + 10 pM CD71mab-saporin, cetuximab-Lys-(dendron(-L-SO1861)4)4,4, and cetuximab-Lys-(dendron(-L-SO1861)4)4,4 + 10 pM CD71mab-saporin in EGFR++ / CD71+ cells (MDA-MB-468). B. Cytotoxic activity of trastuzumab, trastuzumab + 10 pM CD71mab-saporin, trastuzumab-Cys-(dendron(-L-SO1861)4)4, trastuzumab-Cys-(dendron(-L-SO1861)4)4 + 10 pM CD71mab-saporin, trastuzumab-Lys-(dendron(-L-SO1861)4)4,7, or trastuzumab-Lys-(dendron(-L-SO1861)4)4,7 + 10 pM CD71mab-saporin in HER2++ / CD71+ (SK-BR-3) cell line. C. Cytotoxic activity of cetuximab, cetuximab + 10pM CD71mab-saporin, cetuximab-Cys-(dendron(-L-SO1861)4)3,9, cetuximab-Cys-(dendron-L-SO1861)4)3,9 + 10pM CD71mab-saporin, cetuximab-Lys-(dendron(-L-SO1861)4)4,4, or cetuximab-Lys-(dendron(-L-SO1861)4)4,4 + 10pM CD71mab-saporin in EGFR+ / CD71+ cells (CaSki).D. Cytotoxic activity of trastuzumab, trastuzumab + 10 pM CD71mab-saporin, trastuzumab-Cys-(dendron(-L-SO1861)4)4, trastuzumab-Cys-(dendron(-L-SO1861)4)4 + 10 pM CD71mab-saporin, trastuzumab-Lys-(dendron(-L-SO1861)4)4,7, or trastuzumab-Lys-(dendron(-L-SO1861)4)4,7 + 10 pM CD71mab-saporin in HER2+ / - / CD71+ cells (JIMT-1). [Figure 8]Figure 4-1: Legend and axes are the same for Figures A, B, C, and D. A. Cytotoxic activity of cetuximab, cetuximab + 10 pM CD71mab-saporin, cetuximab-Cys-(dendron(-L-SO1861)4)3,9, cetuximab-Cys-(dendron(-L-SO1861)4)3,9 + 10 pM CD71mab-saporin, cetuximab-Lys-(dendron(-L-SO1861)4)4,4, and cetuximab-Lys-(dendron(-L-SO1861)4)4,4 + 10 pM CD71mab-saporin in EGFR++ / CD71+ cells (MDA-MB-468). B. Cytotoxic activity of trastuzumab, trastuzumab + 10 pM CD71mab-saporin, trastuzumab-Cys-(dendron(-L-SO1861)4)4, trastuzumab-Cys-(dendron(-L-SO1861)4)4 + 10 pM CD71mab-saporin, trastuzumab-Lys-(dendron(-L-SO1861)4)4,7, or trastuzumab-Lys-(dendron(-L-SO1861)4)4,7 + 10 pM CD71mab-saporin in HER2++ / CD71+ (SK-BR-3) cell line. C. Cytotoxic activity of cetuximab, cetuximab + 10pM CD71mab-saporin, cetuximab-Cys-(dendron(-L-SO1861)4)3,9, cetuximab-Cys-(dendron-L-SO1861)4)3,9 + 10pM CD71mab-saporin, cetuximab-Lys-(dendron(-L-SO1861)4)4,4, or cetuximab-Lys-(dendron(-L-SO1861)4)4,4 + 10pM CD71mab-saporin in EGFR+ / CD71+ cells (CaSki).D. Cytotoxic activity of trastuzumab, trastuzumab + 10 pM CD71mab-saporin, trastuzumab-Cys-(dendron(-L-SO1861)4)4, trastuzumab-Cys-(dendron(-L-SO1861)4)4 + 10 pM CD71mab-saporin, trastuzumab-Lys-(dendron(-L-SO1861)4)4,7, or trastuzumab-Lys-(dendron(-L-SO1861)4)4,7 + 10 pM CD71mab-saporin in HER2+ / - / CD71+ cells (JIMT-1). [Figure 9] Figure 5-1: Cytotoxic activity of T-DM1, T-DM1 + ​​25.6 nM trastuzumab, or T-DM1 + ​​5.3 nM trastuzumab-Cys-(dendron(-L-SO1861)4)4 in HER2-expressing cells (SK-BR-3). [Figure 10] Figure 6-1: HSP27 gene silencing activity of HSP27BNA-dendron(-L-SO1861)4 compared to HSP27BNA alone. [Figure 11] Figure 7-1: A. Schematic diagram of the release of SO1861 from dendron (-L-SO1861)4 under acidic conditions. [Figure 12] Figure 8-1: Legend and axes are the same for Figures A and B. A. Cell-killing activity of "naked" dendron (Dendron (NEM)4), dendron (NEM)4 + 10 pM EGF-gianthine, dendron (-L-SO1861)4, or dendron (L-SO1861)4 + 10 pM EGF-gianthine in EGFR-expressing A431 cells. B. Cell-killing activity of "naked" dendron (Dendron (NEM)4), dendron (NEM)4 + 10 pM EGF-gianthine, dendron (-L-SO1861)4, or dendron (L-SO1861)4 + 10 pM EGF-gianthine in EGFR-expressing HeLa cells. [Figure 13] Figure 9-1: Legend and axes are the same for Figures A, B, and C. A. Effect of trastuzumab on cell viability of a range of cancer cells. B. Effect of cetuximab on cell viability of a range of cancer cells. C. Effect of T-DM1 on cell viability of a range of cancer cells. [Figure 14] Figure 9-1: Legend and axes are the same for Figures A, B, and C. A. Effect of trastuzumab on cell viability of a range of cancer cells. B. Effect of cetuximab on cell viability of a range of cancer cells. C. Effect of T-DM1 on cell viability of a range of cancer cells. [Figure 15] Figure 10-1: Schematic diagram of monoclonal antibody-(SO1861-scaffold-antisense BNA oligo) conjugate. [Figure 16] Figure 11-1: Schematic of a one-target, two-component system using a monoclonal antibody conjugated to a toxin and the same monoclonal antibody conjugated to a saponin-containing scaffold. [Figure 17] Figure 12-1: Schematic of a two-target, two-component system using a monoclonal antibody conjugated to a toxin and a different monoclonal antibody with a different target conjugated to a saponin-containing scaffold. [Figure 18] Figure 13-1: Schematic diagram of dendron (-L-SO1861)4. [Figure 19] Figure 14-1: Schematic diagram of dendron (-L-SO1861)8. [Figure 20] Figure 15-1: Schematic diagram of SO1861-L-trifunctional linker-L-HSP27BNA. [Figure 21] Figure 16-1: Reaction scheme for the synthesis of dendron (SO1861)4-HSP27BNA oligoconjugate. [Figure 22] Figure 17-1: Model scaffold consisting of four molecular arms for saponin binding via Schiff bases (imines) and one arm for click chemistry. The polymer structure is a first-generation pentavalent polyethylene glycol-based dendrimer. [Figure 23] Figure 18-1: Cell viability of HER14 cells after treatment with pentameric dendrimer (pentrimer), pentrimer in the presence of SA1641, dianthine-EGF, dianthine-EFG in the presence of SA1641, pentrimer in the presence of dianthine-EFG, and pentrimer in the presence of dianthine-EGF and SA1641. [Figure 24]Figure 19-1: SO1861 structure with highlighted chemical groups for conjugation of saponin to the polymer structure to enhance endosomal escape. Highlighted groups are aldehyde (black circle), carboxylic acid (dashed circle), alkene (dashed pentagon), and alcohol (dashed box). [Figure 25] Figure 20-1: A. Standard molecular structure of the SO-1861-EMCH conjugate. Maleimide groups are marked with circles. B. 3D model of the SO1861-EMCH conjugate. Maleimide groups are marked with circles. [Figure 26] Figure 21-1: Reaction scheme for the production of poly(SO1861) using SO1861-EMCH as the monomer, APS / TMEDA system as the polymerization initiator, and aminopropanethiol as the radical quencher. [Figure 27] Figure 22-1: Schematic of the DNA approach. Use of the principles of DNA origami to generate a DNA-based scaffold capable of conjugating and releasing glycoside molecules. In addition, one of the DNA strands carries a click chemistry moiety that can be used for conjugation to a targeted toxin to form a functionalized scaffold. bp: base pair. [Figure 28] Figure 23-1: Schematic of the poly(peptide-SO1861) approach. Use of peptide sequences that can conjugate and release glycoside molecules and react with them to form poly(peptide-SO1861) constructs. The terminus of the poly(peptide) chain can be further modified with click chemistry moieties (e.g., BCN-NHS linkers) that can be used for conjugation to toxins. [Figure 29] Figure 24-1: Molecular structure of a G4-dendron bearing a protected amino group. [Figure 30] Figure 25-1: Schematic of the basic scaffold with click chemistry functional groups for linking any desired effector molecule. [Figure 31]Figure 26-1: Schematic of a functionalized scaffold with a pre-bound effector molecule and click chemistry functional groups for linking any desired ligand. Optionally, a pH-sensitive linker can be provided to release the effector molecule from the scaffold after reaching the endosome. [Figure 32] Figure 1-2: Antibody-protein toxin + unconjugated SO1861 in vivo study. BT474 tumor-bearing mice treated with various concentrations of trastuzumab-saporin (iv) + 1.5 mg / kg unconjugated SO1861 (subQ injection 1 hour before trastuzumab-saporin treatment). [Figure 33] Figure 2-2: Enhanced endosomal escape and target cell killing mediated by unconjugated saponins. A) Cell viability analysis of HeLa cells (EGFR+) treated with SO1861, SO1832, SO1862 (an isomer of SO1861), or SO1904 with or without 1.5 pM EGF dianthine. B) Cell viability analysis of HeLa cells (EGFR+) treated with EGF dianthine and fixed concentrations of SO1861, SO1832, SO1862 (an isomer of SO1861), or SO1904. C) Cell viability analysis of HeLa cells (EGFR+) treated with SO1861 or GE1741 with or without 1.5 pM EGF dianthine. D) Cell viability analysis of HeLa cells (EGFR+) treated with various QS mixes (saponin mixtures from Quillaia Saponaria) with or without 1.5pMEGF dianthine. [Figure 34]Figure 2-2: Enhanced endosomal escape and target cell killing mediated by unconjugated saponins. A) Cell viability analysis of HeLa cells (EGFR+) treated with SO1861, SO1832, SO1862 (an isomer of SO1861), or SO1904 with or without 1.5 pM EGF dianthine. B) Cell viability analysis of HeLa cells (EGFR+) treated with EGF dianthine and fixed concentrations of SO1861, SO1832, SO1862 (an isomer of SO1861), or SO1904. C) Cell viability analysis of HeLa cells (EGFR+) treated with SO1861 or GE1741 with or without 1.5 pM EGF dianthine. D) Cell viability analysis of HeLa cells (EGFR+) treated with various QS mixes (saponin mixtures from Quillaia Saponaria) with or without 1.5pMEGF dianthine. [Figure 35] Figure 3-2: Activity of unconjugated SO1861 versus SO1861-EMCH. EGFR-targeted antisense BNA oligo delivery and gene silencing in cancer cells according to the present invention. A, B, C) Cell viability analysis of A431 (EGFR++), HeLa (EGFR+), or A2058 (EGFR-) cells treated with SO1861 or SO1861-EMCH with or without 1.5 pM EGGF dianthine. D, E) Cell viability analysis of A431 (EGFR++) or HeLa (EGFR+) cells treated with SO1861 or SO1861-N3 with or without 1.5 pM EGGF dianthine. [Figure 36] Figure 3-2: Activity of unconjugated SO1861 versus SO1861-EMCH. EGFR-targeted antisense BNA oligo delivery and gene silencing in cancer cells according to the present invention. A, B, C) Cell viability analysis of A431 (EGFR++), HeLa (EGFR+), or A2058 (EGFR-) cells treated with SO1861 or SO1861-EMCH with or without 1.5 pM EGGF dianthine. D, E) Cell viability analysis of A431 (EGFR++) or HeLa (EGFR+) cells treated with SO1861 or SO1861-N3 with or without 1.5 pM EGGF dianthine. [Figure 37]Figure 3-2: Activity of unconjugated SO1861 versus SO1861-EMCH. EGFR-targeted antisense BNA oligo delivery and gene silencing in cancer cells according to the present invention. A, B, C) Cell viability analysis of A431 (EGFR++), HeLa (EGFR+), or A2058 (EGFR-) cells treated with SO1861 or SO1861-EMCH with or without 1.5 pM EGGF dianthine. D, E) Cell viability analysis of A431 (EGFR++) or HeLa (EGFR+) cells treated with SO1861 or SO1861-N3 with or without 1.5 pM EGGF dianthine. [Figure 38] Figure 4-2: Unconjugated SO1861 vs. SO1861-EMCH (unstable) vs. SO1861-S (stable). Cell viability analysis of HeLa cells (EGFR+) treated with SO1861, SO1861-S (S=HATU, stable linker), and SO1861-EMCH (unstable linker) with or without EGF dianthin. [Figure 39] Figure 5-2: EGFR-targeted antisense BNA oligonucleotide delivery and gene silencing. HSP27 mRNA expression analysis of A431 (EGFR++) and A2058 (EGFR-) cells treated with cetuximab-(Cys-L-SO1861)3.9 or cetuximab-(Cys-L-SO1861)3.9 + 100 nM HSP27 BNA. [Figure 40] Figure 5-2: EGFR-targeted antisense BNA oligonucleotide delivery and gene silencing. HSP27 mRNA expression analysis of A431 (EGFR++) and A2058 (EGFR-) cells treated with cetuximab-(Cys-L-SO1861)3.9 or cetuximab-(Cys-L-SO1861)3.9 + 100 nM HSP27 BNA. [Figure 41] Figure 6-2: Tumor-targeted antisense BNA oligonucleotide delivery and gene silencing in tumor-bearing mice. Mice treated with HSP27 BNA + cetuximab-(Cys-L-SO1861)3,9 in A431 tumor-bearing mice demonstrate efficient tumor-targeted gene silencing compared to controls. [Figure 42] Figure 1-3: HSP27BNA gene silencing activity of HSP27BNA, HSP27BNA-SO1861, and HSP27BNA-Dendron-(SO1861)4 in A431 cancer cell line. [Figure 43] Figure 1-4: Tumor-targeted protein toxin delivery results in tumor volume reduction and tumor growth inhibition in tumor-bearing mice. A. Dose-escalating (intraperitoneal i.p.) administration of cetuximab-(Cys-L-SO1861)3,9(Lys-S-gianthine)2 in A431 tumor-bearing mice demonstrates tumor volume reduction compared to control. B. Dose-escalating intraperitoneal i.p. administration of cetuximab-(Cys-L-SO1861)3,9(Lys-L-gianthine)2 in A431 tumor-bearing mice demonstrates tumor growth reduction compared to control. Intravenous i.v. dose-escalating cetuximab-(Cys-L-SO1861)3,9(Lys-L-gianthine)2 in A431 tumor-bearing mice demonstrates tumor growth reduction compared to control. [Figure 44] Figure 1-4: Tumor-targeted protein toxin delivery results in tumor volume reduction and tumor growth inhibition in tumor-bearing mice. A. Dose-escalating (intraperitoneal i.p.) administration of cetuximab-(Cys-L-SO1861)3,9(Lys-S-gianthine)2 in A431 tumor-bearing mice demonstrates tumor volume reduction compared to control. B. Dose-escalating intraperitoneal i.p. administration of cetuximab-(Cys-L-SO1861)3,9(Lys-L-gianthine)2 in A431 tumor-bearing mice demonstrates tumor growth reduction compared to control. Intravenous i.v. dose-escalating cetuximab-(Cys-L-SO1861)3,9(Lys-L-gianthine)2 in A431 tumor-bearing mice demonstrates tumor growth reduction compared to control. [Figure 45]Figure 2-4: Tumor-targeted antisense BNA oligonucleotide delivery and gene silencing in tumor-bearing mice compared to controls. 30 mg / kg cetuximab-(Cys-L-SO1861)3,9(Lys-L-HSP27BNA)1,8 in A431 tumor-bearing mice demonstrates induced efficient tumor-targeted gene silencing compared to controls. [Figure 46] Figure 3-4: Tumor-targeted antisense BNA oligonucleotide delivery and gene silencing in tumor-bearing mice compared to controls. 30 mg / kg cetuximab-Cys-(SO1861-L-trifunctional linker-L-HSP27BNA)3,7 in A431 tumor-bearing mice demonstrates induced efficient tumor-targeted gene silencing compared to controls. [Figure 47] Figure 4-4: HER2- or EGFR-targeted protein toxin delivery and cell killing in cancer cells according to the present invention. A. SK-BR-3 cells (HER2++) treated with trastuzumab-(Cys-L-SO1861)3,8(Lys-L-gianthine)1,7 or trastuzumab-(Cys-L-SO1861)3,8(Lys-S-gianthine)1,7 and control. B. MDA-MB-468 cells (HER2-) treated with trastuzumab-(Cys-L-SO1861)3,8(Lys-L-gianthine)1,7 or trastuzumab-(Cys-L-SO1861)3,8(Lys-S-gianthine)1,7 and control. C. A431 cells (EGFR++) treated with cetuximab-(Cys-L-SO1861)3,8(Lys-L-gianthine)1,7 or cetuximab-(Cys-L-SO1861)3,8(Lys-S-gianthine)1,7 and control. D. A2058 cells (EGFR-) treated with cetuximab-(Cys-L-SO1861)3,8(Lys-L-gianthine)1,7 or cetuximab-(Cys-L-SO1861)3,8(Lys-S-gianthine)1,7 and control. [Figure 48]Figure 4-4: HER2- or EGFR-targeted protein toxin delivery and cell killing in cancer cells according to the present invention. A. SK-BR-3 cells (HER2++) treated with trastuzumab-(Cys-L-SO1861)3,8(Lys-L-gianthine)1,7 or trastuzumab-(Cys-L-SO1861)3,8(Lys-S-gianthine)1,7 and control. B. MDA-MB-468 cells (HER2-) treated with trastuzumab-(Cys-L-SO1861)3,8(Lys-L-gianthine)1,7 or trastuzumab-(Cys-L-SO1861)3,8(Lys-S-gianthine)1,7 and control. C. A431 cells (EGFR++) treated with cetuximab-(Cys-L-SO1861)3,8(Lys-L-gianthine)1,7 or cetuximab-(Cys-L-SO1861)3,8(Lys-S-gianthine)1,7 and control. D. A2058 cells (EGFR-) treated with cetuximab-(Cys-L-SO1861)3,8(Lys-L-gianthine)1,7 or cetuximab-(Cys-L-SO1861)3,8(Lys-S-gianthine)1,7 and control. [Figure 49]Figure 4-4: HER2- or EGFR-targeted protein toxin delivery and cell killing in cancer cells according to the present invention. A. SK-BR-3 cells (HER2++) treated with trastuzumab-(Cys-L-SO1861)3,8(Lys-L-gianthine)1,7 or trastuzumab-(Cys-L-SO1861)3,8(Lys-S-gianthine)1,7 and control. B. MDA-MB-468 cells (HER2-) treated with trastuzumab-(Cys-L-SO1861)3,8(Lys-L-gianthine)1,7 or trastuzumab-(Cys-L-SO1861)3,8(Lys-S-gianthine)1,7 and control. C. A431 cells (EGFR++) treated with cetuximab-(Cys-L-SO1861)3,8(Lys-L-gianthine)1,7 or cetuximab-(Cys-L-SO1861)3,8(Lys-S-gianthine)1,7 and control. D. A2058 cells (EGFR-) treated with cetuximab-(Cys-L-SO1861)3,8(Lys-L-gianthine)1,7 or cetuximab-(Cys-L-SO1861)3,8(Lys-S-gianthine)1,7 and control. [Figure 50]Figure 4-4: HER2- or EGFR-targeted protein toxin delivery and cell killing in cancer cells according to the present invention. A. SK-BR-3 cells (HER2++) treated with trastuzumab-(Cys-L-SO1861)3,8(Lys-L-gianthine)1,7 or trastuzumab-(Cys-L-SO1861)3,8(Lys-S-gianthine)1,7 and control. B. MDA-MB-468 cells (HER2-) treated with trastuzumab-(Cys-L-SO1861)3,8(Lys-L-gianthine)1,7 or trastuzumab-(Cys-L-SO1861)3,8(Lys-S-gianthine)1,7 and control. C. A431 cells (EGFR++) treated with cetuximab-(Cys-L-SO1861)3,8(Lys-L-gianthine)1,7 or cetuximab-(Cys-L-SO1861)3,8(Lys-S-gianthine)1,7 and control. D. A2058 cells (EGFR-) treated with cetuximab-(Cys-L-SO1861)3,8(Lys-L-gianthine)1,7 or cetuximab-(Cys-L-SO1861)3,8(Lys-S-gianthine)1,7 and control. [Figure 51] Figure 5-4: EGFR-targeted antisense BNA oligo delivery and gene silencing in cancer cells according to the present invention. A. Cetuximab-(Cys-L-SO1861)3,8(Lys-L-HSP27BNA)1,7 treatment and control for A431 cells (EGFR++). B. Cetuximab-(Cys-L-SO1861)3,8(Lys-L-HSP27BNA)1,7 treatment and control for A2058 cells (EGFR-). [Figure 52] Figure 5-4: EGFR-targeted antisense BNA oligo delivery and gene silencing in cancer cells according to the present invention. A. Cetuximab-(Cys-L-SO1861)3,8(Lys-L-HSP27BNA)1,7 treatment and control for A431 cells (EGFR++). B. Cetuximab-(Cys-L-SO1861)3,8(Lys-L-HSP27BNA)1,7 treatment and control for A2058 cells (EGFR-). [Figure 53]Figure 6-4: HER2-targeted antisense BNA oligo delivery and gene silencing in cancer cells according to the present invention. Trastuzumab-(Cys-L-SO1861)3,8(Lys-L-HSP27BNA)3,5 treatment and control in SK-BR-3 cells (HER2++). [Figure 54] Figure 7-4: EGFR-targeted antisense BNA oligo delivery and gene silencing in cancer cells according to the present invention. A. Cetuximab-Cys-(SO1861-L-trifunctional linker-L-HSP27BNA) 3,7 treatment and control in A431 cells (EGFR++). B. Cetuximab-Cys-(SO1861-L-trifunctional linker-L-HSP27BNA) 3,7 treatment and control in A2058 cells (EGFR-). [Figure 55] Figure 8-4: Control treatments for all cell lines. Cell viability when trastuzumab (A), cetuximab (B), T-DM1, (C) free toxins: saporin and dianthin (D), or saporin coupled to non-cell-binding IgG (D) are used as treatments for the indicated cell lines: SK-BR-3, JIMT-1, MDA-MB-468, A431, CaSki, HeLa, A2058, and BT-474. [Figure 56] Figure 8-4: Control treatments for all cell lines. Cell viability when trastuzumab (A), cetuximab (B), T-DM1, (C) free toxins: saporin and dianthin (D), or saporin coupled to non-cell-binding IgG (D) are used as treatments for the indicated cell lines: SK-BR-3, JIMT-1, MDA-MB-468, A431, CaSki, HeLa, A2058, and BT-474. [Figure 57]Figure 9-4: (S)n-(L)(E) Concept: mAb-(SO1861)n (protein toxin)n SO1861 at a cysteine ​​residue (Cys) and a protein toxin (ribosome-inactivating protein) at a lysine residue are both conjugated to the same antibody (mAb) for delivery and internalization into target cells. 1) mAb-(Cys-L-SO1861)4 (Lys-protein toxin)2 binds to its corresponding cell surface receptor, 2) receptor-mediated endocytosis of the conjugate occurs, 3) at low endolysosomal pH and appropriate concentration, SO1861 becomes active, enabling endolysosomal escape, 4) release of the toxin into the cytoplasm occurs, and 5) the toxin induces cell death. [Figure 58] Figure 10-4: (S)n-(L)(E) Concept: mAb-(SO1861)n (antisense BNA oligo)n. Both SO1861 at a cysteine ​​residue (Cys) and an antisense BNA oligonucleotide at a lysine residue are conjugated to the same antibody (mAb) for delivery and internalization into target cells. 1) mAb-(Cys-SO1861)4 (Lys-BNA oligo)2 binds to its corresponding cell surface receptor, 2) receptor-mediated endocytosis of both conjugates occurs, 3) at low endolysosomal pH and appropriate concentrations, SO1861 becomes active, enabling endolysosomal escape, 4) release of the BNA oligo into the cytoplasm, and 5) induction of target gene silencing. [Figure 59] Figure 11-4: (S)n-(L)(E) Concept: mAb-(SO1861-scaffold-antisense BNA oligo)n. (SO1861-trifunctional linker-BNA oligo)n is conjugated to an antibody (mAb) for delivery and internalization into target cells. 1) mAb-(SO1861-trifunctional linker-BNA oligo)4 binds to its corresponding cell surface receptor, 2) receptor-mediated endocytosis of both conjugates occurs, 3) at low endolysosomal pH and appropriate concentrations, SO1861 becomes active and enables endolysosomal escape, 4) release of the BNA oligo into the cytoplasm occurs, and 5) targeted gene silencing is induced. [Figure 60] Figure 12-4: Antibody-SO1861 conjugation procedure. The coupling reaction for the four moieties of the plant-derived saponin SO1861 to the four cysteines on the light chain of an antibody is shown. First, the disulfide bonds on the IgG are blocked under the influence of exposure to TCEP (tris(2-carboxyethyl)phosphine); second, the saponin SO1861 containing a chemical linker attached to it is added along with trifluoroacetic acid, linking the four saponin moieties to the IgG. To produce a cleavable "ready-to-conjugate" saponin, the aldehyde group of SO1861 was reacted with an EMCH (ε-maleimidocaproic acid hydrazide) linker. The hydrazide group of EMCH forms an acid-cleavable hydrazone bond with the aldehyde of SO1861. At the same time, the EMCH linker presents a maleimide group that is thiol (sulfhydryl) reactive and can therefore be conjugated to a thiol of an IgG, i.e., a ligand moiety, thereby providing an endosomal escape-enhancing conjugate of the invention and / or providing a first binding molecule of the invention. [Figure 61] Figure 1-5: In vivo activity of 1T2C. In A431 tumor-bearing mice, the 1T2C combination of 50 mg / kg cetuximab-(Cys-L-SO1861)4 + 25 mg / kg cetuximab-(-L-HSP27BNA)4 demonstrates robust tumor-targeted gene silencing compared to controls. [Figure 62] Figure 2-5: In vivo activity of 1T2C. The 1T2C combination of 40 mg / kg trastuzumab-(Cys-L-SO1861)4 + 0.02 / 0.03 mg / kg trastuzumab-saporin in a PDX tumor mouse model (high HER2 expression) shows effective tumor growth inhibition. [Figure 63]Figure 3-5: One target, two components. EGFR-targeted cell killing in A431 cells (EGFR++) (A, C) and CaSKi cells (EGFR+) (B, D) by therapeutic combinations according to the invention. A, B) Cetuximab-(Cys-L-SO1861) 3,7 titration + fixed concentration of 10 pM cetuximab-saporin and control on A431 (A) and CaSKi (B) cells. C, D) Cetuximab-saporin titration + fixed concentration of 75 nM cetuximab-(Cys-L-SO1861) 3,7 and control on A431 (C) and CaSKi (D) cells. Note: See Table 19 for target receptor expression data (determined by FACS analysis) for each cell line. [Figure 64] Figure 3-5: One target, two components. EGFR-targeted cell killing in A431 cells (EGFR++) (A, C) and CaSKi cells (EGFR+) (B, D) by therapeutic combinations according to the invention. A, B) Cetuximab-(Cys-L-SO1861) 3,7 titration + fixed concentration of 10 pM cetuximab-saporin and control on A431 (A) and CaSKi (B) cells. C, D) Cetuximab-saporin titration + fixed concentration of 75 nM cetuximab-(Cys-L-SO1861) 3,7 and control on A431 (C) and CaSKi (D) cells. Note: See Table 19 for target receptor expression data (determined by FACS analysis) for each cell line. [Figure 65] Figures 4-5: One target, two components. EGFR-targeted cell killing in HeLa cells (EGFR+ / -) (A, C) and A2058 cells (EGFR-) (B, D) by therapeutic combinations according to the invention. A, B) Cetuximab-(Cys-L-SO1861) 3,7 titration + fixed concentration of 10 pM cetuximab-saporin and control on HeLa (A) and CaSKi (B) cells. C, D) Cetuximab-saporin titration + fixed concentration of 75 nM cetuximab-(Cys-L-SO1861) 3,7 and control on HeLa (C) and A2058 (D) cells. Note: See Table 19 for target receptor expression data (determined by FACS analysis) for each cell line. [Figure 66]Figures 4-5: One target, two components. EGFR-targeted cell killing in HeLa cells (EGFR+ / -) (A, C) and A2058 cells (EGFR-) (B, D) by therapeutic combinations according to the invention. A, B) Cetuximab-(Cys-L-SO1861) 3,7 titration + fixed concentration of 10 pM cetuximab-saporin and control on HeLa (A) and CaSKi (B) cells. C, D) Cetuximab-saporin titration + fixed concentration of 75 nM cetuximab-(Cys-L-SO1861) 3,7 and control on HeLa (C) and A2058 (D) cells. Note: See Table 19 for target receptor expression data (determined by FACS analysis) for each cell line. [Figure 67] Figure 5-5: One target, two components. HER2-targeted cell killing in SKBR3 cells (HER2++) (A, B) by therapeutic combinations according to the invention. A) Trastuzumab-(Cys-L-SO1861)4 titration + fixed concentration of 50 pM trastuzumab-saporin and control on SKBR3 cells. B) Trastuzumab-saporin titration + fixed concentration of 2.5 nM trastuzumab-(Cys-L-SO1861)4 and control on SKBR3 cells. Note: See Table 19 for target receptor expression data (determined by FACS analysis) for each cell line. [Figure 68] Figure 6-5: One target, two components. HER2-targeted cell killing in JIMT-1 cells (HER2+ / -) (A, C) and MDA-MB-468 cells (HER2-) (B, D) by therapeutic combinations according to the invention. A, B) Trastuzumab-(Cys-L-SO1861)4 titration + fixed concentration of 50 pM trastuzumab-saporin and control for JIMT-1 (A) and MDA-MB-468 (B) cells. C, D) Trastuzumab-saporin titration + fixed concentration of 2.5 nM trastuzumab-(Cys-L-SO1861)4 and control for JIMT-1 (C) and MDA-MB-468 (D) cells. Note: See Table 19 for target receptor expression data (determined by FACS analysis) for each cell line. [Figure 69]Figure 6-5: One target, two components. HER2-targeted cell killing in JIMT-1 cells (HER2+ / -) (A, C) and MDA-MB-468 cells (HER2-) (B, D) by therapeutic combinations according to the invention. A, B) Trastuzumab-(Cys-L-SO1861)4 titration + fixed concentration of 50 pM trastuzumab-saporin and control for JIMT-1 (A) and MDA-MB-468 (B) cells. C, D) Trastuzumab-saporin titration + fixed concentration of 2.5 nM trastuzumab-(Cys-L-SO1861)4 and control for JIMT-1 (C) and MDA-MB-468 (D) cells. Note: See Table 19 for target receptor expression data (determined by FACS analysis) for each cell line. [Figure 70] Figure 7-5: Chloroquine inhibits one target, two components. HER2- and EGFR-targeted cell killing in SK-BR-3 (HER2++) and A431 cells (EGFR++) by therapeutic combinations according to the invention. A) Trastuzumab-saporin titration + fixed concentration of 5 nM trastuzumab-(Cys-L-SO1861)4 + 0.5 μM chloroquine and control for SK-BR-3 cells. B) Cetuximab-saporin titration + fixed concentration of 5 nM cetuximab-(Cys-L-SO1861)3,8 + 0.5 μM chloroquine and control for A431 cells. Note: See Table 19 for target receptor expression data (determined by FACS analysis) for each cell line. [Figure 71]Figure 8-5: One target, two components. EGFR-targeted gene silencing in A431 cells (EGFR++) and A2058 cells (EGFR-) by therapeutic combinations according to the invention. A, B) Cetuximab-(Cys-L-SO1861)3,8 titration + fixed concentration of 100 nM cetuximab-(Lys-L-HSP27BNA)4 and control for A431 cells (A) and A2058 cells (B). C, D) Cetuximab-(Lys-L-HSP27BNA)4 titration + fixed concentration of 77 nM cetuximab-(Cys-L-SO1861)3,8 and control for A431 cells (C) and A2058 cells (D). Note: See Table 19 for target receptor expression data (determined by FACS analysis) for each cell line. [Figure 72] Figure 8-5: One target, two components. EGFR-targeted gene silencing in A431 cells (EGFR++) and A2058 cells (EGFR-) by therapeutic combinations according to the invention. A, B) Cetuximab-(Cys-L-SO1861)3,8 titration + fixed concentration of 100 nM cetuximab-(Lys-L-HSP27BNA)4 and control for A431 cells (A) and A2058 cells (B). C, D) Cetuximab-(Lys-L-HSP27BNA)4 titration + fixed concentration of 77 nM cetuximab-(Cys-L-SO1861)3,8 and control for A431 cells (C) and A2058 cells (D). Note: See Table 19 for target receptor expression data (determined by FACS analysis) for each cell line. [Figure 73]Figure 9-5: Two targets, two components. A) EGFR- and HER2-targeted cell killing in MDA-MB-468 cells (EGFR++) and HeLa cells (EGFR+ / -), and HER2-targeted cell killing in SK-BR-3 cells (HER2++) and JIMT-1 cells (HER2+ / -) by therapeutic combinations according to the invention. A) Cetuximab-Cys-(dendron(-L-SO1861)4)3,9 titration + fixed concentration 10 pM cetuximab-saporin and control in MDA-MB-468 cells (A) and HeLa cells (B). Trastuzumab-Cys-(dendron-L-SO1861)4)4 titration + fixed concentration 50 pM trastuzumab-saporin and control in SK-BR-3 cells (C) and JIMT-1 cells (D). Note: See Table 19 for target receptor expression data (determined by FACS analysis) for each cell line. [Figure 74] Figure 9-5: Two targets, two components. A) EGFR- and HER2-targeted cell killing in MDA-MB-468 cells (EGFR++) and HeLa cells (EGFR+ / -), and HER2-targeted cell killing in SK-BR-3 cells (HER2++) and JIMT-1 cells (HER2+ / -) by therapeutic combinations according to the invention. A) Cetuximab-Cys-(dendron(-L-SO1861)4)3,9 titration + fixed concentration 10 pM cetuximab-saporin and control in MDA-MB-468 cells (A) and HeLa cells (B). Trastuzumab-Cys-(dendron-L-SO1861)4)4 titration + fixed concentration 50 pM trastuzumab-saporin and control in SK-BR-3 cells (C) and JIMT-1 cells (D). Note: See Table 19 for target receptor expression data (determined by FACS analysis) for each cell line. [Figure 75]Figure 10-5: One target, two components. SK-BR-3 cells (HER2+ / -) can be efficiently killed by the therapeutic combination trastuzumab-saporin + 2.5 nM trastuzumab-(Cys-L-SO1861)4 according to the invention, but titration of T-DM1 + ​​2.5 nM trastuzumab-(Cys-L-SO1861)4 is not effective at such low toxin concentrations. T-DM1 is trastuzumab-emtansine (Kadcyla®) and carries 3.5 emtansine (DM1) toxin molecules per antibody (DAR 3.5). Note: See Table 19 for target receptor expression data (determined by FACS analysis) for each cell line. [Figure 76] Figure 11-5: Control treatment of all cell lines. A-D) Cell viability when treated with the indicated cell lines SK-BR-3, JIMT-1, MDA-MB-468, A431, CaSki, HeLa, A2058, and BT-474 with trastuzumab (A), cetuximab (B), T-DM1 (C), free toxins: saporin and giantin (D), or saporin coupled to non-cell-binding IgG (D). Note: See Table 19 for target receptor expression data (determined by FACS analysis) for each cell line. [Figure 77] Figure 11-5: Control treatment of all cell lines. A-D) Cell viability when treated with the indicated cell lines SK-BR-3, JIMT-1, MDA-MB-468, A431, CaSki, HeLa, A2058, and BT-474 with trastuzumab (A), cetuximab (B), T-DM1 (C), free toxins: saporin and giantin (D), or saporin coupled to non-cell-binding IgG (D). Note: See Table 19 for target receptor expression data (determined by FACS analysis) for each cell line. [Figure 78]Figure 12-5: One target, two components. EGFR-targeted cell killing in A431 cells (EGFR++) (A), CaSKi cells (EGFR+) (B), and A2058 cells (EGFR-) by therapeutic combinations according to the invention. A, B, C) Cetuximab-(Cys-L-QS mix) 4,1 titration + fixed concentration of 10 pM cetuximab-saporin or 10 pM cetuximab-giantin and control in A431 cells (A), CaSKi cells (B), and A2058 cells (C). QS mix is ​​a mixture of saponins from the extract Quillaja Saponaria. Note: See Table 19 for target receptor expression data (determined by FACS analysis) for each cell line. [Figure 79] Figure 12-5: One target, two components. EGFR-targeted cell killing in A431 cells (EGFR++) (A), CaSKi cells (EGFR+) (B), and A2058 cells (EGFR-) by therapeutic combinations according to the invention. A, B, C) Cetuximab-(Cys-L-QS mix) 4,1 titration + fixed concentration of 10 pM cetuximab-saporin or 10 pM cetuximab-giantin and control in A431 cells (A), CaSKi cells (B), and A2058 cells (C). QS mix is ​​a mixture of saponins from the extract Quillaja Saponaria. Note: See Table 19 for target receptor expression data (determined by FACS analysis) for each cell line. [Figure 80] Figure 13-5: One-target, two-component concept: mAb1-SO1861 + mAb1-protein toxin. SO1861 and a toxin (ribosome-inactivating protein) are each independently conjugated to an antibody (mAb1) for delivery and internalization into target cells. 1) mAb1-SO1861 and mAb1-protein toxin bind to cell surface receptors, 2) receptor-mediated endocytosis of both conjugates occurs, 3) at low endolysosomal pH and appropriate concentrations, SO1861 becomes active and enables endolysosomal escape, 4) release of the toxin into the cytoplasm occurs, and 5) the toxin induces cell death. [Figure 81]Figure 14-5: One-target, two-component concept: mAb1-SO1861 + mAb2-BNA oligo. SO1861 and antisense BNA oligonucleotides are each independently conjugated to an antibody (mAb1) for delivery and internalization into target cells. 1) mAb1-SO1861 and mAb1-BNA oligo bind to cell surface receptors, 2) receptor-mediated endocytosis of both conjugates occurs, 3) at low endolysosomal pH and appropriate concentrations, SO1861 becomes active and enables endolysosomal escape, 4) release of the BNA oligo into the cytoplasm, and 5) target gene silencing occurs. [Figure 82] Figure 15-5: One-target, two-component concept: mAb1-(scaffold(-SO1861)n)n + mAb1-protein toxin. The dendron (-SO1861)n and protein toxin (ribosome-inactivating protein) are each independently conjugated to an antibody (mAb1) for delivery and internalization into target cells. 1) mAb1-dendron (-SO1861)4 and mAb1-protein toxin bind to cell surface receptors, 2) receptor-mediated endocytosis of both conjugates occurs, 3) at low endolysosomal pH and appropriate concentrations, SO1861 becomes active and enables endolysosomal escape, 4) release of the toxin into the cytoplasm occurs, and 5) the toxin induces cell death. [Figure 83] Figure 1-6: The 2T2 component system tested in an A431 tumor-bearing mouse model reveals tumor regression. [Figure 84] Figure 2-6: The 2T2 component system tested in an A431 tumor-bearing mouse model demonstrates tumor regression and eradication. [Figure 85]Figures 3-6: Two targets, two components. EGFR / HER2-targeted cell killing in A431 cells (EGFR++ / HER2+ / -) (A, C) and CaSKi cells (EGFR++ / HER2+ / -) (B, D) by therapeutic combinations according to the invention. A, B) Cetuximab-(Cys-L-SO1861)3,7 titration + fixed concentration of 50 pM trastuzumab-saporin and control for A431 cells. C, D) Trastuzumab-saporin titration + fixed concentration of 75 nM cetuximab-(Cys-L-SO1861)3,7 and control for CaSKi cells. Legend and / or axes are the same for all A, B, C, or D. [Figure 86] Figures 3-6: Two targets, two components. EGFR / HER2-targeted cell killing in A431 cells (EGFR++ / HER2+ / -) (A, C) and CaSKi cells (EGFR++ / HER2+ / -) (B, D) by therapeutic combinations according to the invention. A, B) Cetuximab-(Cys-L-SO1861)3,7 titration + fixed concentration of 50 pM trastuzumab-saporin and control for A431 cells. C, D) Trastuzumab-saporin titration + fixed concentration of 75 nM cetuximab-(Cys-L-SO1861)3,7 and control for CaSKi cells. Legend and / or axes are the same for all A, B, C, or D. [Figure 87] Figures 4-6: Two targets, two components. EGFR / HER2-targeted cell killing in HeLa cells (EGFR+ / - / HER2+ / -) (A, C) and A2058 cells (EGFR- / HER2+ / -) (B, D) by therapeutic combinations according to the invention. A, B) Cetuximab-(Cys-L-SO1861)3,7 titration + fixed concentration of 50 pM trastuzumab-saporin and control for HeLa cells. C, D) Trastuzumab-saporin titration + fixed concentration of 75 nM cetuximab-(Cys-L-SO1861)3,7 and control for A2058 cells. Legend and / or axes are the same for all A, B, C, or D. [Figure 88]Figures 4-6: Two targets, two components. EGFR / HER2-targeted cell killing in HeLa cells (EGFR+ / - / HER2+ / -) (A, C) and A2058 cells (EGFR- / HER2+ / -) (B, D) by therapeutic combinations according to the invention. A, B) Cetuximab-(Cys-L-SO1861)3,7 titration + fixed concentration of 50 pM trastuzumab-saporin and control for HeLa cells. C, D) Trastuzumab-saporin titration + fixed concentration of 75 nM cetuximab-(Cys-L-SO1861)3,7 and control for A2058 cells. Legend and / or axes are the same for all A, B, C, or D. [Figure 89] Figures 5-6: Two targets, two components. HER2 / EGFR-targeted cell killing in SKBR3 cells (HER2++ / EGFR+ / -) (A, B) by a therapeutic combination according to the invention. A) Trastuzumab-(Cys-L-SO1861)4 titration + fixed concentration of 1.5 pM EGF dianthine and control on SKBR3 cells. B) EGF dianthine titration + fixed concentration of 2.5 nM trastuzumab-(Cys-L-SO1861)4 and control on SKBR3 cells. [Figure 90] Figure 6-6: Two targets, two components. HER2 / EGFR-targeted cell killing in JIMT-1 cells (HER2+ / -EGFR+ / -) (A, C) and MDA-MB-468 cells (HER2- / EGFR++) (B, D) by therapeutic combinations according to the invention. A, B) Trastuzumab-(Cys-L-SO1861)4 titration + fixed concentration of 1.5 pM EGF dianthine and control for JIMT-1 cells. C, D) EGF dianthine titration + fixed concentration of 2.5 nM trastuzumab-(Cys-L-SO1861)4 and control for MDA-MB-468 cells. Legend and / or axes are the same for all A, B, C, or D. [Figure 91]Figure 6-6: Two targets, two components. HER2 / EGFR-targeted cell killing in JIMT-1 cells (HER2+ / -EGFR+ / -) (A, C) and MDA-MB-468 cells (HER2- / EGFR++) (B, D) by therapeutic combinations according to the invention. A, B) Trastuzumab-(Cys-L-SO1861)4 titration + fixed concentration of 1.5 pM EGF dianthine and control for JIMT-1 cells. C, D) EGF dianthine titration + fixed concentration of 2.5 nM trastuzumab-(Cys-L-SO1861)4 and control for MDA-MB-468 cells. Legend and / or axes are the same for all A, B, C, or D. [Figure 92] Figure 7-6: Two targets, two components. HER2 / EGFR-targeted cell killing in SKBR3 cells (HER2++ / EGFR+ / -) (A, B) by a therapeutic combination according to the invention. A) Trastuzumab-(Cys-L-SO1861)4 titration + fixed concentration of 10 pM cetuximab-saporin and control on SKBR3 cells. B) Cetuximab-saporin titration + fixed concentration of 2.5 nM trastuzumab-(Cys-L-SO1861)4 and control on SKBR3 cells. [Figure 93] Figure 8-6: Two targets, two components. HER2 / EGFR-targeted cell killing in JIMT-1 cells (HER2+ / -EGFR+ / -) (A, C) and MDA-MB-468 cells (HER2- / EGFR++) (B, D) by therapeutic combinations according to the invention. A, B) Trastuzumab-(Cys-L-SO1861)4 titration + fixed concentration of 10 pM cetuximab-saporin and control for JIMT-1 cells. C, D) Cetuximab-saporin titration + fixed concentration of 2.5 nM trastuzumab-(Cys-L-SO1861)4 and control for MDA-MB-468 cells. Legend and / or axes are the same for all A, B, C, or D. [Figure 94]Figure 8-6: Two targets, two components. HER2 / EGFR-targeted cell killing in JIMT-1 cells (HER2+ / -EGFR+ / -) (A, C) and MDA-MB-468 cells (HER2- / EGFR++) (B, D) by therapeutic combinations according to the invention. A, B) Trastuzumab-(Cys-L-SO1861)4 titration + fixed concentration of 10 pM cetuximab-saporin and control for JIMT-1 cells. C, D) Cetuximab-saporin titration + fixed concentration of 2.5 nM trastuzumab-(Cys-L-SO1861)4 and control for MDA-MB-468 cells. Legend and / or axes are the same for all A, B, C, or D. [Figure 95] Figure 9-6: Chloroquine inhibits two targets, two components. GFR / HER2, EGFR / CD71, or HER2 / CD71-targeted cell killing in EA431 cells (EGFR++ / HER2+ / - / CD71+) (A, B), MDA-MB-468 cells (EGFR++ / HER2- / CD71+) (C), or SK-BR-3 (HER2++ / EGFR+ / - / CD71+) (D) by a therapeutic combination according to the invention + chloroquine. A) Trastuzumab-giantine or trastuzumab-saporin titration + a fixed concentration of 75 nM cetuximab-(Cys-L-SO1861)3,9 + 800 nM chloroquine, and control, on A431 cells. B) CD71mab-saporin titration + fixed concentration of 10.5 nM cetuximab-(Cys-L-SO1861)3,9 + 500 nM chloroquine and control on A431 cells. C) CD71mab-saporin titration + fixed concentration of 10.5 nM cetuximab-(Cys-L-SO1861)3,9 + 500 nM chloroquine and control on MDA-MB-468 cells. D) CD71mab-saporin titration + fixed concentration of 5 nM trastuzumab-(Cys-L-SO1861)3,9 + 500 nM chloroquine and control on SK-BR-3 cells. [Figure 96]Figure 9-6: Chloroquine inhibits two targets, two components. GFR / HER2, EGFR / CD71, or HER2 / CD71-targeted cell killing in EA431 cells (EGFR++ / HER2+ / - / CD71+) (A, B), MDA-MB-468 cells (EGFR++ / HER2- / CD71+) (C), or SK-BR-3 (HER2++ / EGFR+ / - / CD71+) (D) by a therapeutic combination according to the invention + chloroquine. A) Trastuzumab-giantine or trastuzumab-saporin titration + a fixed concentration of 75 nM cetuximab-(Cys-L-SO1861)3,9 + 800 nM chloroquine, and control, on A431 cells. B) CD71mab-saporin titration + fixed concentration of 10.5 nM cetuximab-(Cys-L-SO1861)3,9 + 500 nM chloroquine and control on A431 cells. C) CD71mab-saporin titration + fixed concentration of 10.5 nM cetuximab-(Cys-L-SO1861)3,9 + 500 nM chloroquine and control on MDA-MB-468 cells. D) CD71mab-saporin titration + fixed concentration of 5 nM trastuzumab-(Cys-L-SO1861)3,9 + 500 nM chloroquine and control on SK-BR-3 cells. [Figure 97] Figure 10-6: Two targets, two components. EGFR / HER2-targeted gene silencing in A431 cells (EGFR++ / HER2+ / -) (A) and A2058 cells (EGFR- / HER2+ / -) (B) by a therapeutic combination according to the invention. A) Cetuximab-(Cys-L-SO1861)3,9 titration + a fixed concentration of 100 nM trastuzumab-(Lys-L-HSP27BNA)4,4, and control for A431 cells (A) and A2058 cells (B). C, D) Trastuzumab-(Lys-HSP27BNA)4,4 titration + a fixed concentration of 77 nM cetuximab-(Cys-L-SO1861)3,9, and control for A431 cells (A) and A2058 cells (B). The legend and / or axes are the same for all A, B, C, or D. [Figure 98]Figure 10-6: Two targets, two components. EGFR / HER2-targeted gene silencing in A431 cells (EGFR++ / HER2+ / -) (A) and A2058 cells (EGFR- / HER2+ / -) (B) by a therapeutic combination according to the invention. A) Cetuximab-(Cys-L-SO1861)3,9 titration + a fixed concentration of 100 nM trastuzumab-(Lys-L-HSP27BNA)4,4, and control for A431 cells (A) and A2058 cells (B). C, D) Trastuzumab-(Lys-HSP27BNA)4,4 titration + a fixed concentration of 77 nM cetuximab-(Cys-L-SO1861)3,9, and control for A431 cells (A) and A2058 cells (B). The legend and / or axes are the same for all A, B, C, or D. [Figure 99] Figure 11-6: Two targets, two components. A) EGFR / CD71 or HER2 / CD71 targeted cell killing in MDA-MB-468 cells (EGFR++ / CD71+) (A) HeLa cells (EGFR+ / - / CD71+), SK-BR-3 cells (HER2++ / CD71+) (B), and JIMT-1 cells (HER2+ / - / CD71+) by therapeutic combinations according to the invention. A) Cetuximab-Cys-(Dendron(-L-SO1861)4)3,9 titration + fixed concentration 10 pM CD71mab-saporin and control on MDA-MB-468 cells. B) A) Cetuximab-Cys-(Dendron(-L-SO1861)4)3,9 titration + fixed concentration 10 pM CD71mab-saporin and control on HeLa cells. C) Trastuzumab-Cys-(dendron(-L-SO1861)4)4 titration + fixed concentration 10 pM CD71mab-saporin and control on SK-BR-3 cells. D) Trastuzumab-Cys-(dendron(-L-SO1861)4)4 titration + fixed concentration 10 pM CD71mab-saporin and control on JIMT-1 cells. [Figure 100]Figure 11-6: Two targets, two components. A) EGFR / CD71 or HER2 / CD71 targeted cell killing in MDA-MB-468 cells (EGFR++ / CD71+) (A) HeLa cells (EGFR+ / - / CD71+), SK-BR-3 cells (HER2++ / CD71+) (B), and JIMT-1 cells (HER2+ / - / CD71+) by therapeutic combinations according to the invention. A) Cetuximab-Cys-(Dendron(-L-SO1861)4)3,9 titration + fixed concentration 10 pM CD71mab-saporin and control on MDA-MB-468 cells. B) A) Cetuximab-Cys-(Dendron(-L-SO1861)4)3,9 titration + fixed concentration 10 pM CD71mab-saporin and control on HeLa cells. C) Trastuzumab-Cys-(dendron(-L-SO1861)4)4 titration + fixed concentration 10 pM CD71mab-saporin and control on SK-BR-3 cells. D) Trastuzumab-Cys-(dendron(-L-SO1861)4)4 titration + fixed concentration 10 pM CD71mab-saporin and control on JIMT-1 cells. [Figure 101] Figure 12-6: Two targets, two components versus T-DM1. A431 cells (EGFR++ / HER2+ / -) can be efficiently killed by the therapeutic combination trastuzumab-saporin + 75 nM cetuximab-(Cys-L-SO1861)3,9 according to the invention, but titration of T-DM1 + ​​75 nM cetuximab-(Cys-L-SO1861)3,9 is not effective at such low toxin concentrations. T-DM1 is trastuzumab emtansine (Kadcyla®) and carries 3.5 emtansine (DM1) toxin molecules per antibody. [Figure 102]Figure 13-6: Two targets, two components. EGFR / CD71 and EGFR / HER2 targeted cell killing in A431 cells (EGFR+++ / HER2+ / -) (A), CaSKi cells (EGFR++ / HER2+ / -) (B), and A2058 cells (EGFR- / HER2+ / -) by a therapeutic combination according to the invention. A, B, C) Cetuximab-(Cys-L-QS mix) 4,1 titration + fixed concentration of 10 pM trastuzumab-saporin or 10 pM CD71mab-saporin, and control, on A431 cells (A), CaSKi cells (B), and A2058 cells (C). QS mix is ​​a mixture of saponins from the extract Quillaja Saponaria. [Figure 103] Figure 13-6: Two targets, two components. EGFR / CD71 and EGFR / HER2 targeted cell killing in A431 cells (EGFR+++ / HER2+ / -) (A), CaSKi cells (EGFR++ / HER2+ / -) (B), and A2058 cells (EGFR- / HER2+ / -) by a therapeutic combination according to the invention. A, B, C) Cetuximab-(Cys-L-QS mix) 4,1 titration + fixed concentration of 10 pM trastuzumab-saporin or 10 pM CD71mab-saporin, and control, on A431 cells (A), CaSKi cells (B), and A2058 cells (C). QS mix is ​​a mixture of saponins from the extract Quillaja Saponaria. [Figure 104] Figure 14-6: Control treatment of all cell lines. A-D) Cell viability when treated with trastuzumab (A), cetuximab (B), T-DM1 (C), free toxins: saporin and dianthin (D), or saporin coupled to non-cell-binding IgG (D) indicated cell lines SK-BR-3, JIMT-1, MDA-MB-468, A431, CaSki, HeLa, A2058, BT-474. Legend and / or axes are the same for all A, B, C, or D. [Figure 105]Figure 14-6: Control treatment of all cell lines. A-D) Cell viability when treated with trastuzumab (A), cetuximab (B), T-DM1 (C), free toxins: saporin and dianthin (D), or saporin coupled to non-cell-binding IgG (D) indicated cell lines SK-BR-3, JIMT-1, MDA-MB-468, A431, CaSki, HeLa, A2058, BT-474. Legend and / or axes are the same for all A, B, C, or D. [Figure 106] Figure 14-6: Control treatment of all cell lines. A-D) Cell viability when treated with trastuzumab (A), cetuximab (B), T-DM1 (C), free toxins: saporin and dianthin (D), or saporin coupled to non-cell-binding IgG (D) indicated cell lines SK-BR-3, JIMT-1, MDA-MB-468, A431, CaSki, HeLa, A2058, BT-474. Legend and / or axes are the same for all A, B, C, or D. [Figure 107] Figure 15-6: Two-target, two-component concept: mAb1-SO1861 + mAb2-protein toxin. SO1861 and a toxin (ribosome-inactivating protein) are each separately conjugated to an antibody (mAb) for delivery and internalization into target cells. 1) mAb1-SO1861 and mAb2-protein toxin bind to their corresponding cell surface receptors, 2) receptor-mediated endocytosis of both conjugates occurs, 3) at low endolysosomal pH and appropriate concentrations, SO1861 becomes active and enables endolysosomal escape, 4) release of the toxin into the cytoplasm occurs, and 5) the toxin induces cell death. [Figure 108]Figure 16-6: Two-target, two-component concept: mAb1-SO1861 + mAb2-BNA oligo. SO1861 and antisense BNA oligonucleotides are each separately conjugated to an antibody (mAb) for delivery and internalization into target cells. 1) mAb1-SO1861 and mAb2-BNA oligo bind to their corresponding cell surface receptors, 2) receptor-mediated endocytosis of both conjugates occurs, 3) at low endolysosomal pH and appropriate concentrations, SO1861 becomes active and enables endolysosomal escape, 4) release of the BNA oligo into the cytoplasm, and 5) target gene silencing occurs. [Figure 109] Figure 17-6: [Figure 110] Figure 1-7: EGFR-targeted gene silencing in A431 cells (EGFR++) (A) and A2058 cells (EGFR-) (B) by a combination of unconjugated SO1861-EMCH and HSP27BNA. Also, SO1861-EMCH and cetuximab-(Lys-L-HSP27BNA) titration + fixed concentration of 4000 nM SO1861-EMCH and control in A431 cells (A) and A2058 cells (B). (A) and (B) are graphs based on HSP27BNA or cetuximab-HSP27BNA conjugate (nM). EGFR-targeted gene silencing in A431 cells (EGFR++) (A) and A2058 cells (EGFR-) (B) by unconjugated SO1861-EMCH with free HSP27BNA, and SO1861-EMCH, and conjugated cetuximab-(Lys-L-HSP27BNA) titration + fixed concentration 4000 nM SO1861-EMCH, and control in A431 cells (C) and A2058 cells (D). (C) and (D) are graphs based on HSP27BNA (nM). The legends for Figures 1-7B and D also apply to Figures 1-7A and C. [Figure 111]Figure 1-7: EGFR-targeted gene silencing in A431 cells (EGFR++) (A) and A2058 cells (EGFR-) (B) by a combination of unconjugated SO1861-EMCH and HSP27BNA. Also, SO1861-EMCH and cetuximab-(Lys-L-HSP27BNA) titration + fixed concentration of 4000 nM SO1861-EMCH and control in A431 cells (A) and A2058 cells (B). (A) and (B) are graphs based on HSP27BNA or cetuximab-HSP27BNA conjugate (nM). EGFR-targeted gene silencing in A431 cells (EGFR++) (A) and A2058 cells (EGFR-) (B) by unconjugated SO1861-EMCH with free HSP27BNA, and SO1861-EMCH, and conjugated cetuximab-(Lys-L-HSP27BNA) titration + fixed concentration 4000 nM SO1861-EMCH, and control in A431 cells (C) and A2058 cells (D). (C) and (D) are graphs based on HSP27BNA (nM). The legends for Figures 1-7B and D also apply to Figures 1-7A and C. DETAILED DESCRIPTION OF THE INVENTION

[0103] For a bioactive molecule to work, it must be able to engage its target, for example, in serum, on the cell surface, or within a cell or organelle. The active moiety of nearly all protein-based targeted toxins must enter the cytosol of the target cell to mediate its target-modulating effect. In many constellations, the toxin remains ineffective because (1) the targeting moiety is poorly internalized and remains bound to the outside of the cell, (2) it is recycled to the cell surface after internalization, or (3) it is transported to the endolysosome, where it is degraded. These fundamental issues have been known for decades, and more than 500 targeted toxins have been investigated over the past several decades. However, the problem remains unresolved, and only one antibody-targeted protein toxin, moxetumomab pasudotox-tdfk (LUMOXITI®, AstraZeneca Pharmaceuticals LP), has been approved by the FDA to date for relapsed or refractory hairy cell leukemia.

[0104] Many strategies to overcome these problems have been described, including approaches to target toxins to endogenous membrane transport complexes of the biosynthetic pathway in the endoplasmic reticulum and techniques to block or weaken the membrane integrity of endosomes, i.e., compartments of the endocytic pathway in cells, thereby facilitating endosomal escape. These include the use of lysosomotropic amines, carboxylic acid ionophores, calcium channel antagonists, various cell-penetrating peptides of viral, bacterial, plant, animal, human, and synthetic origin, other organic molecules, and light-inducible techniques. While the efficacy of targeted toxins has typically been increased by 100- or 1,000-fold in cell culture, and in exceptional cases by more than 1,000,000-fold, the requirement to coadminister endosomal escape-enhancing agents with other substances introduces new challenges, including additional side effects, loss of target specificity, difficulty in determining the therapeutic window, and cell-type-dependent variability.

[0105] All strategies involving physicochemical techniques require enhancer molecules that interact more or less directly with membranes and essentially include small chemical molecules, secondary metabolites, peptides, and proteins. A common feature of all these substances is that they are not target cell specific themselves and distribute with kinetics other than targeted toxins. This is one major drawback of current approaches.

[0106] While the present invention will be described with reference to specific embodiments, the invention is not limited thereto but only by the claims. The embodiments of the invention described herein can work in combination and in concert, unless otherwise specified.

[0107] While the present invention has been described in terms of several embodiments, it is contemplated that alternatives, modifications, permutations, and equivalents thereof will become apparent to those skilled in the art upon reading and understanding the specification and upon studying the drawings and graphs. The present invention is in no way limited to the illustrated embodiments. Changes may be made without departing from the scope defined by the appended claims.

[0108] One aspect of the present invention relates to a therapeutic drug molecule having the chemical structure of Compound I: A1 m ((-L9 w )((-L1 q -B1 n ) u ((-L2 r -L3 s )(-L4 v -C) p ) t )) x (Compound I)

[0109] wherein A1 is a first ligand when B1 is a first effector moiety, or A1 is a first effector moiety when B1 is a first ligand; C is saponin; When A1 is a first ligand and B1 is a first effector moiety, then m=0 or 1; When A1 is a first effector moiety and B1 is a first ligand, m=0 to 32; When B1 is a first ligand and A1 is a first effector moiety, or when A1 is a first ligand and B1 is a first effector moiety, n=0 or 1; p = 1 to 128; L1 is at least one linker for covalently coupling two chemical groups; L2 is at least one linker for covalently coupling two chemical groups; L3 is at least one oligomeric or polymeric backbone for covalently coupling two chemical groups; L4 is at least one linker for covalently coupling two chemical groups; L9 is a trifunctional linker for covalently coupling three chemical groups; q=0 or 1; r=0 or 1; s=0 or 1; If s=0, t=0, 1, or 2, and if s=1, t=0 to 16; u=0-32 when A1 is a first ligand and B1 is a first effector moiety, or u=1 when A1 is a first effector moiety and B1 is a first ligand; v=0 or 1; w=1 or 0; x=1~16.

[0110] One embodiment of the invention relates to a therapeutic combination comprising a therapeutic molecule according to the invention and a second therapeutic molecule having the chemical structure of Compound II: A2 a ((-L10 j )((-L5 d -B2 b ) h ((-L6 e -L7 f )(-L8i -C) c ) g )) k (Compound II)

[0111] wherein A2 is a second ligand when B2 is a second effector moiety, or A2 is a second effector moiety when B2 is a second ligand; C is saponin; a=0 or 1 when A2 is a second ligand and B2 is a second effector moiety, or a=0-32 when A2 is a second effector moiety and B2 is a second ligand; b=0 or 1 when B2 is a second ligand and A2 is a second effector moiety, or when A2 is a second ligand and B2 is a second effector moiety; c = 1 to 128; L5 is at least one linker for covalently coupling two chemical groups; L6 is at least one linker for covalently coupling two chemical groups; L7 is at least one oligomeric or polymeric backbone for covalently coupling two chemical groups; L8 is at least one linker for covalently coupling two chemical groups; L10 is a trifunctional linker for covalently coupling three chemical groups; d=0 or 1; e=0 or 1; f=0 or 1; When f=0, g=0, 1, 2, when f=1, g=0 to 16; h=0-32 when A2 is a second ligand and B2 is a second effector moiety, or h=1 when A2 is a second effector moiety and B2 is a second ligand; i=0 or 1; j=1 or 0; k=1 to 16.

[0112] One embodiment is a second therapeutic molecule of the invention, where g=0, 1, or 2 when f=0 and t>0, g=1 or 2 when f=0 and t=0, g=0-16 when f=1 and t>0, or g=1-16 when f=1 and t=0.

[0113] One embodiment is a therapeutic molecule of the invention or a second therapeutic molecule of the invention, wherein the first ligand A1 or B1 and / or the second ligand A2 or B2 comprises or consists of an immunoglobulin, an immunoglobulin binding domain or an immunoglobulin binding fragment, such as an antibody, IgG, a molecule comprising or consisting of a Vhh domain or a Vh domain, a Fab, scFv, Fv, dAb, F(ab)2, Fcab fragment, or comprises or consists of at least one non-proteinaceous ligand and / or at least one proteinaceous ligand, a ligand for binding to a cell surface molecule such as EGF or a cytokine, wherein the first ligand and second ligand are the same or different.

[0114] In one embodiment is a therapeutic molecule of the invention or a second therapeutic molecule of the invention, wherein the first ligand A1 or B1 and / or the second ligand A2 or B2 is preferably selected from the group consisting of CD71, CA125, EpCAM(17-1A), CD52, CEA, CD44v6, FAP, EGF-IR, integrin, syndecan-1, vascular integrin alpha-V beta-3, HER2, EGFR, CD20, CD22, folate receptor 1, CD146, CD56, CD19, CD138, CD27L receptor, PSMA, CanAg, integrin-alpha V, CA6, CD33, mesothelin, Cripto, CD3, CD30, CD239, CD70, CD123, CD352, DLL3, CD25, ephrin A4, MUC1, Trop2, CEACAM5, CEACAM6, HER3, C The first ligand binds to a tumor cell epitope, preferably a tumor cell-specific epitope, on a tumor cell receptor, preferably a tumor cell-specific receptor, selected from D74, PTK7, Notch3, FGF2, C4.4A, FLT3, CD38, FGFR3, CD7, PD-L1, CTLA4, CD52, PDGFRA, VEGFR1, VEGFR2, more preferably CD71, EGFR, and HER2, with the proviso that the first ligand and the second ligand bind to the same or different tumor cell epitopes, preferably tumor cell-specific epitopes, and / or the tumor cell receptor, preferably the tumor cell-specific receptor, to which the first ligand can bind is the same as or different from the tumor cell receptor, preferably the tumor cell receptor, preferably the tumor cell-specific receptor, to which the second ligand can bind.

[0115] In one embodiment is a therapeutic molecule of the invention or a second therapeutic molecule of the invention, wherein the first ligand A1 or B1 and / or the second ligand A2 or B2 is selected from the group consisting of cetuximab, daratumumab, gemtuzumab, trastumumab, panitumumab, brentuximab, inotuzumab, moxetumomab, polatuzumab, obinutuzumab, OKT-9 anti-CD71 monoclonal antibody in IgG form, pertuzumab, rituximab, ofatumumab, herceptin, alemtuzumab, pi The antibody comprises or consists of nutuzumab, OKT-10 anti-CD38 monoclonal antibody, an antibody of Table A2 or Table A3 or Table A4, preferably cetuximab or trastuzumab or OKT-9, or at least one tumor cell receptor binding domain and / or at least one tumor cell receptor binding fragment thereof, which are preferably tumor cell-specific receptor binding domain(s) and / or tumor cell-specific receptor binding fragment(s), with the proviso that the first ligand is the same as or different from the second ligand.

[0116] One embodiment is a therapeutic molecule of the invention or a second therapeutic molecule of the invention, wherein the first ligand A1 or B1 is internalized by the tumor cell following binding of the first ligand to its binding partner on the tumor cell, preferably wherein binding of the first ligand to the tumor cell is followed by tumor cell receptor-mediated internalization, e.g., via endocytosis, of the complex of the first ligand and the binding partner of the first ligand on the tumor cell.

[0117] One embodiment is a therapeutic drug molecule of the invention or a second therapeutic drug molecule of the invention, wherein the second ligand A2 or B2 is internalized by the tumor cell following binding of the second ligand to its binding partner on the tumor cell, preferably wherein binding of the second ligand to the tumor cell is followed by tumor cell receptor-mediated internalization, e.g., via endocytosis, of the complex of the second ligand and the binding partner of the second ligand on the tumor cell.

[0118] In one embodiment, the therapeutic molecule of the invention or the second therapeutic molecule of the invention, wherein the first effector moiety A1 or B1 and / or the second effector moiety A2 or B2 comprises or consists of any one or more of an oligonucleotide, a nucleic acid, and a xenonucleic acid, preferably a vector, a gene, a transgene that induces cell suicide, deoxyribonucleic acid (DNA), ribonucleic acid (RNA), antisense oligonucleotide (ASO, AON), short interfering RNA (siRNA), microRNA (miRNA), a DNA aptamer, an RNA aptamer, mRNA, minicircle DNA, peptide nucleic acid (PNA), phosphoroa The effector moiety is selected from any one or more of midate morpholino oligomer (PMO), locked nucleic acid (LNA), bridged nucleic acid (BNA), 2'-deoxy-2'-fluoroarabinonucleic acid (FANA), 2'-O-methoxyethyl-RNA (MOE), 2'-O,4'-aminoethylene-bridged nucleic acid, 3'-fluorohexitol nucleic acid (FHNA), plasmid, glycol nucleic acid (GNA), and threose nucleic acid (TNA), or a derivative thereof, more preferably a BNA, for example a BNA for silencing HSP27 protein expression, with the proviso that the first effector moiety and the second effector moiety are the same or different.

[0119] In one embodiment, the therapeutic molecule of the invention or the second therapeutic molecule of the invention, wherein the first effector moiety A1 or B1 and / or the second effector moiety A2 or B2 comprises or consists of at least one proteinaceous molecule, preferably selected from any one or more of peptides, proteins, enzymes such as urease and Cre recombinase, proteinaceous toxins, ribosome-inactivating proteins, at least one protein toxin selected from Table A5, and / or bacterial toxins, plant toxins, more preferably viral toxins, such as apoptin; bacterial toxins, such as Shiga toxins, Shiga-like toxins, Pseudomonas The protein toxin is selected from any one or more of: P. aeruginosa exotoxin (PE) or exotoxin A of PE, full-length or truncated diphtheria toxin (DT), cholera toxin; fungal toxins such as alpha-sarcin; plant toxins including ribosome-inactivating proteins and the A chain of type 2 ribosome-inactivating proteins, such as giantin, e.g., giantin-30 or giantin-32, saporin, e.g., saporin-S3 or saporin-S6, bouganin or a deimmunized derivative of bouganin, debouganin, Shiga-like toxin A, pokeweed antiviral protein, ricin, ricin A chain, modeccin, modeccin A chain, abrin, abrin A chain, volkensin, volkensin A chain, viscumin, viscumin A chain; or animal or human toxins such as frog RNase, granzyme B, or angiogenin from humans, or any fragment or derivative thereof. Preferably, the protein toxin is giantin and / or saporin. However, the first effector moiety(s) and the second effector moiety(s) may be the same or different.

[0120] An embodiment is a therapeutic molecule of the invention or a second therapeutic molecule of the invention, wherein the first effector moiety A1 or B1 and / or the second effector moiety A2 or B2 comprises or consists of at least one payload, preferably one or more of a ribosome-targeting toxin, an elongation factor-targeting toxin, a tubulin-targeting toxin, a DNA-targeting toxin, and an RNA-targeting toxin, more preferably emtansine, pasdotox, maytansinoid derivative DM1, maytansinoid derivative DM4, monomethyl auristatin E (MMAE, vedotin), monomethyl auristatin F (MM AF, mafodotin), calicheamicin, N-acetyl-γ-calicheamicin, pyrrolobenzodiazepine (PBD) dimer, benzodiazepine, CC-1065 analog, duocarmycin, doxorubicin, paclitaxel, docetaxel, cisplatin, cyclophosphamide, etoposide, docetaxel, 5-fluorouracil (5-FU), mitoxantrone, tubulysin, indolinobenzodiazepine, AZ13599185, cryptophycin, rhizoxin, methotrexate, anthracyclines, camptothecin analog, SN-38, DX-8951f, exatecan mesylate, Pseudomonas The steroid inhibitor is selected from one or more of a truncated form of P. aeruginosa exotoxin (PE38), a duocarmycin derivative, amanitin, α-amanitin, splicostatin, tylanstatin, ozogamicin, tesirin, amberstatin 269, and soravtansine, or a derivative thereof.

[0121] In one embodiment is a therapeutic molecule of the invention or a second therapeutic molecule of the invention, wherein the therapeutic molecule and / or second therapeutic molecule comprises or consists of any one of gemtuzumab ozogamicin, brentuximab vedotin, trastuzumab emtansine, inotuzumab ozogamicin, moxetumomab pasudotox, and polatuzumab vedotin, and the antibody-drug conjugates of Table A2 and Table A3, or at least one tumor cell-specific receptor binding domain thereof and / or at least one tumor cell-specific receptor binding fragment thereof, which are preferably tumor cell-specific receptor binding domain(s) and / or tumor cell-specific receptor binding fragment(s), with the proviso that the therapeutic molecule and the second therapeutic molecule are the same or different.

[0122] In one embodiment is a therapeutic molecule of the invention or a second therapeutic molecule of the invention, wherein saponin C is a triterpenoid saponin or a bisdesmosidic triterpene saponin belonging to the 12,13-dehydrooleanane type having an aldehyde functional group at position C-23 and optionally a glucuronic acid functional group in the carbohydrate substituent of the C-3 beta-OH group of the saponin, and / or a saponin isolated from Gypsophila species and / or Saponaria species and / or Agrostemma species and / or Quillaja species, e.g., Quillaja saponaria.

[0123] In one embodiment is a therapeutic molecule of the invention or a second therapeutic molecule of the invention, wherein saponin C is a single specific saponin or a mixture of two or more different saponins, e.g., one or more of the saponins in Table A1 or Scheme I, SO1861, SA1657, GE1741, SA1641, QS-21, QS-21A, QS-21 A-api, QS-21 A-xyl, QS-21B, QS-21 B-api, QS-21 B-xyl, QS-7-xyl, QS-7-api, QS-17-api, QS-17-xyl, QS1861, QS1862, Quillajasaponin, Saponinum album, QS-18, Quil-A, Gyp1, gypsoside A, AG1, AG2, SO1542, SO1584, SO1658, SO1674, SO1832, or any of their stereoisomers and / or any combination thereof. Preferably, the saponin is SO1861 and / or GE1741 and / or SA1641 and / or QS-21 and / or a saponin having a quillaric acid aglycone core, a Gal-(1→2)-[Xyl-(1→3)]-GlcA carbohydrate substituent at the C-3 beta-OH group, and a Glc-(1→3)-Xyl-(1→4)-Rha-(1→2)-[Xyl-(1→3)-4-OAc-Qui-(1→4)]-Fuc carbohydrate substituent at the C-28-OH group. and / or 3-O-beta-D-galactopyranosyl-(1→2)-[beta-D-xylopyranosyl-(1→3)]-beta-D-glucuronopyranosylquilayic acid, 28-O-beta-D-glucopyranosyl-(1→3)-beta-D-xylopyranosyl-(1→4)-alpha-L-rhamnopyranosyl-(1→2)-[beta-D-xylopyranosyl-(1→3)-4-OAc-beta-D-quinovopyranosyl-(1→4)]-beta-D-fucopyranoside. More preferably, the saponin is SO1861 and / or QS-21.

[0124] An embodiment is a therapeutic molecule of the invention or a second therapeutic molecule of the invention, wherein saponin C is a bisdesmosidic saponin and has a molecular mass of at least 1,500 daltons, comprises an oleanane-type triterpene containing an aldehyde group at the C-23 position and optionally a hydroxyl group at the C-16 position, has a first branched carbohydrate side chain at the C-3 position, which first branched carbohydrate side chain optionally contains glucuronic acid, and the saponin contains an ester group with a second branched carbohydrate side chain at the C-28 position, which second branched carbohydrate chain preferably contains at least four carbohydrate units, and optionally contains at least one acetyl residue, e.g., two acetyl residues, and / or at least one or at least one saponin is selected from the group consisting of QS-21, QS-21A, QS-21B, QS-21C, QS-21D, QS-21E, QS-21F, QS-21G, QS-21H ... One or more of A-api, QS-21 A-xyl, QS-21B, QS-21 B-api, QS-21 B-xyl, QS-7-xyl, QS-7-api, QS-17-api, QS-17-xyl, QS-18, QS-1861, protonated QS1861 (QS1862), and Quil-A.

[0125] One embodiment is a therapeutic drug molecule of the invention or a second therapeutic drug molecule of the invention, wherein saponin C is a bisdesmosidic triterpene saponin belonging to the 12,13-dehydroanane type having an aldehyde functional group at position C-23, and saponin C is covalently coupled to an amino acid residue of a first ligand A1 or B1 and / or a first effector moiety B1 or A1 and / or a second ligand A2 or B2 and / or a second effector moiety B2 or A2 via the aldehyde functional group on saponin C, preferably said aldehyde functional group at position C-23, preferably via linkers L2, L4, L6, L8, L9, and / or L10, more preferably via cleavable linkers L2, L4, L6, L8, L9, and / or L10, and wherein the amino acid residue is preferably selected from cysteine ​​and lysine.

[0126] One embodiment is a therapeutic drug molecule of the invention or a second therapeutic drug molecule of the invention, wherein saponin C is a bisdesmosidic triterpene saponin belonging to the 12,13-dehydroanane type having an aldehyde functional group at position C-23, and wherein the aldehyde functional group at position C-23 of at least one saponin is covalently coupled to a linker N-ε-maleimidocaproic acid hydrazide, which is covalently coupled via a thio-ether bond to a sulfhydryl group, e.g., a sulfhydryl group of cysteine, on a first ligand A1 or B1 and / or a first effector moiety B1 or A1 and / or a second ligand A2 or B2 and / or a second effector moiety B2 or A2.

[0127] One embodiment is a therapeutic drug molecule of the invention or a second therapeutic drug molecule of the invention, wherein saponin C is a bisdesmosidic triterpene saponin belonging to the 12,13-dehydrooleanane type having an aldehyde functional group at position C-23 and including a glucuronic acid functional group on the carbohydrate substituent at the C-3 beta-OH group of the saponin, and wherein saponin C is covalently coupled to an amino acid residue of a first ligand A1 or B1 and / or a first effector moiety B1 or A1 and / or a second ligand A2 or B2 and / or a second effector moiety B2 or A2 via the glucuronic acid functional group on saponin C, if present, preferably via linkers L2, L4, L6, L8, L9, and / or L10. The amino acid residue is preferably selected from cysteine ​​and lysine, more preferably the amino acid residue is lysine.

[0128] In one embodiment is a therapeutic agent molecule of the invention or a second therapeutic agent molecule of the invention, wherein saponin C is a bisdesmosidic triterpene saponin belonging to the 12,13-dehydrooleanane type having an aldehyde functional group at position C-23 and comprising a glucuronic acid functional group on a carbohydrate substituent at the C-3 beta-OH group of the saponin, wherein at least one of the glucuronic acid functional groups on the carbohydrate substituent at the C-3 beta-OH group of the saponin is linked to the linker 1-(bis(dimethylamino)methylene)-1H-1,2,3-triazolo[4,5-b]pyridinium 3-fluorouracil. The linker is covalently coupled to a phosphate, which in turn is covalently coupled via an amide bond to an amine group on the first ligand A1 or B1 and / or the first effector moiety B1 or A1 and / or the second ligand A2 or B2 and / or the second effector moiety B2 or A2, for example, to a lysine or N-terminal amine group of the first ligand A1 or B1 and / or the first effector moiety B1 or A1 and / or the second ligand A2 or B2 and / or the second effector moiety B2 or A2.

[0129] One embodiment is a therapeutic drug molecule of the invention or a second therapeutic drug molecule of the invention, wherein the first ligand A1 or B1 and / or the first effector moiety B1 or A1 and / or the second ligand A2 or B2 and / or the second effector moiety B2 or A2 comprises one or more covalently attached saponin C, preferably 2, 3, 4, 5, 6, 8, 10, 16, 32, 64, 128, or 1-100 saponins, or any number of saponins in between, e.g., 7, 9, 12 saponins.

[0130] An embodiment is a therapeutic drug molecule of the invention or a second therapeutic drug molecule of the invention, wherein the first ligand A1 or B1 and / or the first effector moiety B1 or A1 and / or the second ligand A2 or B2 and / or the second effector moiety B2 or A2 comprises one or more covalently attached saponin(s) C. The saponin(s) C is / are attached to the first ligand A1 or B1 and / or the first effector moiety B1 or A1 and / or the second ligand A2 or B2 and / or the second effector moiety B2 or A2 (where r, s, v, e, f, and i are 0), preferably to a cysteine ​​and / or a lysine, either directly or via at least one linker L2, L4, L6, L8, L9, and / or L10, or via at least one cleavable linker L2, L4, L6 , L8, L9, and / or L10 and / or via at least one oligomeric or polymeric backbone L3 and / or L7, preferably 1 to 8 such backbones or 2 to 4 such backbones, wherein at least one backbone is optionally based on a dendron, and 1 to 32 saponins, preferably 2, 3, 4, 5, 6, 8, 10, 16, 32 saponins, or any number of saponins in between, e.g., 7, 9, 12 saponins, are covalently attached to the at least one backbone.

[0131] One embodiment is a therapeutic drug molecule of the invention or a second therapeutic drug molecule of the invention, wherein saponin C is a bisdesmosidic triterpene saponin belonging to the 12,13-dehydroanane type having an aldehyde functional group at position C-23, and saponin C is covalently coupled to an amino acid residue of a first ligand A1 or B1 and / or a first effector moiety B1 or A1 and / or a second ligand A2 or B2 and / or a second effector moiety B2 or A2 via the aldehyde functional group on saponin C, preferably said aldehyde functional group at position C-23, preferably via linkers L2, L4, L6, L8, L9, and / or L10, more preferably via cleavable linkers L2, L4, L6, L8, L9, and / or L10, and wherein the amino acid residue is preferably selected from cysteine ​​and lysine. The cleavable linkers L2, L4, L6, L8, L9, and / or L10 undergo cleavage under acidic, reductive, enzymatic, or photoinduced conditions, preferably the cleavable linker comprises a hydrazone or hydrazide bond that undergoes cleavage under acidic conditions when bound to a saponin, and / or comprises a bond that is susceptible to proteolysis, for example proteolysis by cathepsin B, when bound to a saponin, and / or the cleavable linker comprises a disulfide bond that is susceptible to cleavage under reductive conditions.

[0132] One embodiment is a therapeutic drug molecule of the invention or a second therapeutic drug molecule of the invention, wherein saponin C is a bisdesmosidic triterpene saponin belonging to the 12,13-dehydroanane type having an aldehyde functional group at position C-23, and saponin C is covalently coupled to an amino acid residue of a first ligand A1 or B1 and / or a first effector moiety B1 or A1 and / or a second ligand A2 or B2 and / or a second effector moiety B2 or A2 via the aldehyde functional group on saponin C, preferably said aldehyde functional group at position C-23, preferably via linkers L2, L4, L6, L8, L9, and / or L10, more preferably via cleavable linkers L2, L4, L6, L8, L9, and / or L10, and wherein the amino acid residue is preferably selected from cysteine ​​and lysine. Cleavable linkers L2, L4, L6, L8, L9, and / or L10 undergo cleavage in vivo under acidic conditions present in endosomes and / or lysosomes of mammalian cells, preferably human cells, preferably at a pH between 4.0 and 6.5, more preferably at a pH ≦5.5.

[0133] In one embodiment is a therapeutic drug molecule of the invention or a second therapeutic drug molecule of the invention, wherein the polymer or oligomer backbone L3 and / or L7 comprises a polymer or oligomer structure and comprises chemical groups, chemical groups for covalent coupling of the polymer or oligomer backbone L3 and / or L7 to amino acid residues of a first ligand and / or a first effector moiety and / or a second ligand and / or a second effector moiety.

[0134] In one embodiment is a therapeutic agent molecule of the invention or a second therapeutic agent molecule of the invention, wherein saponin C is a bisdesmosidic triterpene saponin belonging to the 12,13-dehydrooleanane type having an aldehyde functional group at position C-23 and comprising a glucuronic acid functional group on a carbohydrate substituent at the C-3 beta-OH group of the saponin, wherein at least one of the glucuronic acid functional groups on the carbohydrate substituent at the C-3 beta-OH group of the saponin is linked to the linker 1-(bis(dimethylamino)methylene)-1H-1,2,3-triazolo[4,5-b]pyridinium 3-fluorouracil. The linker is covalently coupled to a phosphate group on the first ligand A1 or B1 and / or the first effector moiety B1 or A1 and / or the second ligand A2 or B2 and / or the second effector moiety B2 or A2, for example, a lysine or N-terminal amine group on the first ligand A1 or B1 and / or the first effector moiety B1 or A1 and / or the second ligand A2 or B2 and / or the second effector moiety B2 or A2, via an amide bond. At least one saponin is covalently attached to the polymeric or oligomeric structure of backbone L3 and / or L7 via cleavable linkers L4 and / or L8 according to the invention.

[0135] One embodiment is a therapeutic drug molecule of the invention or a second therapeutic drug molecule of the invention, wherein the chemical group of the polymer or oligomer backbone L3 and / or L7 for covalently coupling the backbone to an amino acid residue of the first ligand and / or first effector moiety and / or second ligand and / or second effector moiety is a click chemistry group, preferably selected from a tetrazine, an azide, an alkene, or an alkyne, or cyclic derivatives of these groups; more preferably, the click chemistry group is an azide.

[0136] One embodiment is a therapeutic drug molecule of the invention or a second therapeutic drug molecule of the invention, wherein the first ligand A1 or B1 and / or the first effector moiety B1 or A1 and / or the second ligand A2 or B2 and / or the second effector moiety B2 or A2 comprises one or more covalently attached saponin C, preferably 2, 3, 4, 5, 6, 8, 10, 16, 32, 64, 128, or 1-100 saponins, or any number of saponins in between, e.g., 7, 9, 12 saponins. The at least one saponin is covalently attached to the first ligand and / or first effector moiety and / or second ligand and / or second effector moiety either directly or via at least one linker such as, for example, a bifunctional linker based on N-ε-maleimidocaproic acid hydrazide and / or based on 1-[bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium 3-oxide hexafluorophosphate, or a trifunctional linker L9 when w=1 or a trifunctional linker L10 when j=1, e.g., the trifunctional linkers of Scheme II.

[0137] In one embodiment is a therapeutic molecule of the invention encompassing the previous embodiment, or a second therapeutic molecule of the invention encompassing the previous embodiment, wherein the trifunctional linker L9 when w=1 and / or the trifunctional linker L10 when j=1 comprises a second chemical group to which at least one saponin is covalently attached, a third chemical group for covalently attaching to a first and / or second ligand, and a first chemical group for covalently attaching to at least one first and / or second effector moiety, preferably the trifunctional linker is the trifunctional linker of Scheme II.

[0138] One embodiment is a therapeutic drug molecule of the invention or a second therapeutic drug molecule of the invention, wherein at least one saponin is covalently attached to a first ligand and / or a first effector moiety and / or a second ligand and / or a second effector moiety via at least one linker comprising a trifunctional linker L9 when j=1 and / or a trifunctional linker L10 when w=1, to which both the first ligand and at least one first effector moiety are attached, and / or to which a second ligand and at least one second effector moiety are attached, preferably wherein the trifunctional linker is a trifunctional linker of Scheme II.

[0139] In one embodiment is a therapeutic drug molecule of the invention or a second therapeutic drug molecule of the invention, wherein the polymeric or oligomeric structure of backbone L3 and / or L7 comprises a linear, branched, and / or cyclic polymer, oligomer, dendrimer, dendron, dendronized polymer, dendronized oligomer, DNA, polypeptide, polylysine, polyethylene glycol, or an assembly of these polymeric or oligomeric structures, which assembly is preferably assembled by covalent crosslinking.

[0140] One embodiment is a therapeutic drug molecule of the invention or a second therapeutic drug molecule of the invention, wherein the first ligand A1 or B1 and / or the first effector moiety B1 or A1 and / or the second ligand A2 or B2 and / or the second effector moiety B2 or A2 comprise one or more covalently linked saponin C, preferably 2, 3, 4, 5, 6, 8, 10, 16, 32, 64, 128, or 1-100 saponins, or any number therebetween, e.g., 7, 9, 12 saponins. The first ligand A1 or B1 is covalently linked to the first effector moiety B1 or A1, respectively, via at least one linker L1, and / or the second ligand A2 or B2 is covalently linked to the second effector moiety B2 or A2, respectively, via at least one linker L5.

[0141] In one embodiment is a therapeutic combination of the invention, wherein the first ligand A1 is a monoclonal antibody or at least one binding fragment or domain thereof according to any one of claims 3 to 7, m=1, q=0, n=0, u=0, r=0, L3 is a scaffold according to the invention, s=1, or L3 is absent and s=0, L4 is a linker or cleavable linker according to the invention, v=1, p=2 to 4, and if s=1 then t=2 to 4, and if s=0 then t=0, saponin C is a saponin according to the invention, preferably saponin C is SO1861 and / or QS-21, and effector moiety A2 is an effector moiety selected from any one or more of an oligonucleotide, a nucleic acid, and a xenonucleic acid, and preferably a vector, a gene, a transgene that induces cell suicide, deoxyribonucleic acid (DNA), ribonucleic acid (RNA), or the like. ), antisense oligonucleotides (ASO, AON), short interfering RNA (siRNA), microRNA (miRNA), DNA aptamer, RNA aptamer, mRNA, minicircle DNA, peptide nucleic acid (PNA), phosphoramidate morpholino oligomer (PMO), locked nucleic acid (LNA), bridged nucleic acid (BNA), 2'-deoxy-2'-fluoroarabinonucleic acid (FANA), 2'-O-methoxyethyl-RNA (MOE), 2'-O,4'-aminoethylene-bridged nucleic acid, 3'-fluorohexitol nucleic acid (FHNA), plasmid, glycol nucleic acid (GNA), and threose nucleic acid (TNA), or derivatives thereof, more preferably BNA, for example, BNA for silencing HSP27 protein expression, wherein a=1, d=0, b=0, h=0, e=0, f=0, i=0, c=0, g=0.

[0142] In one embodiment is a therapeutic molecule of the invention, wherein r=0, s=0, v=0, s=0, v=0, p=0, t=0, the ligand A1 is a monoclonal antibody or at least one binding fragment or domain thereof according to any one of the inventions, and m=1, and the effector moiety B1 is an effector moiety selected from any one or more of an oligonucleotide, a nucleic acid, and a xenonucleic acid, preferably a vector, a gene, a transgene that induces cell suicide, a deoxyribonucleic acid (DNA), a ribonucleic acid (RNA), an antisense oligonucleotide (ASO, AON), a short interfering RNA (siRNA), a microRNA (miRNA), a DNA aptamer, an RNA aptamer, an mRNA, a minicircle DNA, a peptide the linker L1 is selected from one or more of a nucleotide sequence selected from the group consisting of nucleotides of 2'-diamino-2'-fluoroarabino nucleic acid (PNA), phosphoramidate morpholino oligomer (PMO), locked nucleic acid (LNA), bridged nucleic acid (BNA), 2'-deoxy-2'-fluoroarabino nucleic acid (FANA), 2'-O-methoxyethyl-RNA (MOE), 2'-O,4'-aminoethylene-bridged nucleic acid, 3'-fluorohexitol nucleic acid (FHNA), plasmid, glycol nucleic acid (GNA), and threose nucleic acid (TNA), or derivatives thereof, more preferably BNA, for example, BNA for silencing HSP27 protein expression, wherein q=0, n=1, and u=2 to 4, or q=1, n=2 to 4, u=2 to 4, and the linker L1 is an oligomeric or polymeric backbone L3 according to the present invention.

[0143] One embodiment is a therapeutic combination of the invention, wherein the therapeutic molecule is a therapeutic molecule of the previous embodiment, the second ligand A2 is a monoclonal antibody in accordance with the invention or at least one binding fragment or domain thereof, a=1, d=0, b=0, h=0, e=0, L7 is a scaffold in accordance with the invention and f=1 or L7 is absent and f=0, L8 is a linker or cleavable linker in accordance with the invention and i=1, c=2-4, when f=1 g=2-4, when f=0 g=0, and saponin C is a saponin in accordance with the invention, preferably SO1861 and / or QS-21, and wherein ligand A1 and ligand A2 are the same or different.

[0144] Certain aspects of the present invention relate to therapeutic combinations, comprising: (a) a first pharmaceutical composition comprising a therapeutic molecule having the chemical structure of Compound I in accordance with the present invention, the first pharmaceutical composition optionally further comprising a pharmaceutically acceptable excipient; and (b) a second pharmaceutical composition comprising a second therapeutic molecule having the chemical structure of Compound II in accordance with the present invention, the second pharmaceutical composition optionally further comprising a pharmaceutically acceptable excipient.

[0145] One aspect of the present invention relates to a first pharmaceutical composition of the present invention for use as a medicament.

[0146] One aspect of the present invention relates to a therapeutic combination for use in treating or preventing cancer in a human subject, the therapeutic combination comprising (a) a first pharmaceutical composition of the present invention and (b) a second pharmaceutical composition of the present invention, wherein ligand A1 or B1 and ligand A2 or B2 are capable of binding to a tumor cell epitope, preferably a tumor cell-specific epitope, on a tumor cell surface molecule, preferably on a tumor cell-specific surface molecule, with the proviso that the tumor cell epitope or tumor cell-specific epitope to which ligand A1 or B1 can bind is the same as or different from the tumor cell epitope or tumor cell-specific epitope to which ligand A2 or B2 can bind.

[0147] One aspect of the present invention relates to a first pharmaceutical composition of the present invention for use in treating or preventing cancer in a patient in need thereof, wherein the ligand A1 or B1 is capable of binding to a tumor cell epitope, preferably a tumor cell-specific epitope, on a tumor cell surface molecule, preferably a tumor cell-specific surface molecule.

[0148] In one embodiment is a first pharmaceutical composition for use in accordance with the invention or a combination of therapeutic agents for use in accordance with the invention, wherein a second pharmaceutical composition of the invention and a first pharmaceutical composition of the invention are administered to a patient in need thereof.

[0149] One aspect of the present invention relates to a first pharmaceutical composition of the present invention, further comprising a second therapeutic agent molecule of the present invention.

[0150] One aspect of the present invention relates to a first pharmaceutical composition of the present invention, further comprising a second therapeutic drug molecule of the present invention, for use as a medicament.

[0151] One aspect of the present invention relates to a first pharmaceutical composition of the present invention, further comprising a second therapeutic molecule of the present invention, for use in treating or preventing cancer in a human subject.

[0152] An aspect of the present invention of the first series of aspects and embodiments of the present invention relates to a scaffold suitable for covalently attaching at least one biologically active molecule to a carrier molecule, wherein the scaffold comprises a polymeric or oligomeric structure, and wherein at least one of said biologically active molecules is covalently attached to said polymeric or oligomeric structure, and wherein the scaffold further comprises a first chemical group for covalent coupling of the scaffold to the carrier molecule.

[0153] An embodiment of the first series of aspects and embodiments of the present invention is a scaffold according to the present invention, wherein the at least one biologically active molecule has a molecular mass of 3,000 daltons or less, preferably 2,500 daltons or less, more preferably 2,300 daltons or less, most preferably 2,000 daltons or less, for example between 1,700 and 1,950 daltons.

[0154] An embodiment of the first series of aspects and embodiments of the present invention is a scaffold according to the present invention, wherein the at least one biologically active molecule is an amphiphilic molecule.

[0155] One embodiment of the first series of aspects and embodiments of the present invention is a scaffold according to the present invention, wherein when more than one biologically active molecule is covalently attached to the polymeric or oligomeric structure comprised by the scaffold, at least one biologically active molecule is a single specific molecule or a mixture of different molecules.

[0156] One embodiment of the first series of aspects and embodiments of the present invention is a scaffold according to the present invention, wherein at least one biologically active molecule is a glycoside, preferably a bisdesmosidic triterpene or triterpenoid saponin, more preferably a bisdesmosidic triterpene saponin, most preferably a bisdesmosidic triterpene saponin belonging to the 12,13-dehydrooleanane type having an aldehyde functionality at position C-23 and optionally a glucuronic acid functionality on the carbohydrate substituent at the C-3 beta-OH group of the saponin.

[0157] An embodiment of the first series of aspects and embodiments of the present invention is a scaffold according to the present invention, wherein the at least one bioactive molecule is a saponin that may be isolated from Gypsophila species and / or Saponaria species and / or Agrostemma species and / or Quillaja species, such as Quillaja saponaria. Alternatively, it may be a single specific saponin or a mixture of two or more different saponins, for example, one or more of the saponins of Table A1 or Scheme I, SO1861, SA1657, GE1741, SA1641, QS-21, QS-21A, QS-21 A-api, QS-21 A-xyl, QS-21B, QS-21 B-api, QS-21 B-xyl, QS-7-xyl, QS-7-api, QS-17-api, QS-17-xyl, QS1861, QS1862, Quillaja saponin, Saponinum albumenum album, QS-18, Quil-A, Gyp1, gypsoside A, AG1, AG2, SO1542, SO1584, SO1658, SO1674, SO1832, or any stereoisomer thereof and / or any combination thereof. Preferably, the saponin is SO1861 and / or GE1741 and / or SA1641 and / or QS-21, and / or a saponin having a quillaic acid aglycone core, a Gal-(1→2)-[Xyl-(1→3)]-GlcA carbohydrate substituent at the C-3 beta-OH group, and a Glc-(1→3)-Xyl-(1→4)-Rha-(1→2)-[Xyl-(1→3)-4-OAc-Qui-(1→4)]-Fuc carbohydrate substituent at the C-28-OH group. and / or 3-O-beta-D-galactopyranosyl-(1→2)-[beta-D-xylopyranosyl-(1→3)]-beta-D-glucuronopyranosylquilayic acid, 28-O-beta-D-glucopyranosyl-(1→3)-beta-D-xylopyranosyl-(1→4)-alpha-L-rhamnopyranosyl-(1→2)-[beta-D-xylopyranosyl-(1→3)-4-OAc-beta-D-quinovopyranosyl-(1→4)]-beta-D-fucopyranoside. More preferably, the saponin is SO1861 and / or QS-21.

[0158] One embodiment of this first series of aspects and embodiments of the present invention is a scaffold according to the present invention, wherein the at least one biologically active molecule is a bisdesmosidic saponin, having a molecular mass of at least 1,500 daltons, comprising an oleanane-type triterpene containing an aldehyde group at the C-23 position and optionally a hydroxyl group at the C-16 position, and a first branched carbohydrate side chain at the C-3 position, which optionally contains glucuronic acid. The saponin contains an ester group with a second branched carbohydrate side chain at the C-28 position, this second branched carbohydrate chain preferably containing at least four carbohydrate units, optionally containing at least one acetyl residue, for example two acetyl residues, and / or optionally containing a deoxy carbohydrate, and / or optionally containing quinovose, and / or optionally containing glucose, and / or optionally containing 4-methoxycinnamic acid, and / or optionally containing 5-O-[5-O-Ara / Api-3,5-dihydroxy-6-methyl-octanoyl]-3,5-dihydroxy-6-methyl-octanoic acid, and / or optionally containing 5-O-[5-O-Rha-(1→2)-Ara / Api-3,5-dihydroxy-6-methyl-octanoyl]-3,5-dihydroxy-6-methyl-octanoic acid, linked to the carbohydrate via an ester bond. Alternatively, the at least one saponin is QS-21, or QS-21A, QS-21 One or more of A-api, QS-21 A-xyl, QS-21B, QS-21 B-api, QS-21 B-xyl, QS-7-xyl, QS-7-api, QS-17-api, QS-17-xyl, QS-18, QS-1861, protonated QS1861 (QS1862), and Quil-A.

[0159] An embodiment of the first series of aspects and embodiments of the present invention is a scaffold according to the present invention, wherein at least one biologically active molecule is covalently attached to the polymeric or oligomeric structure via a non-cleavable bond or via a cleavable bond, preferably said cleavable bond is susceptible to cleavage under acidic, reductive, enzymatic or photoinduced conditions, more preferably the cleavable bond is a hydrazone or hydrazide bond that is susceptible to cleavage under acidic conditions, and / or a bond that is susceptible to proteolysis, e.g. proteolysis by cathepsin B, and / or a bond that is susceptible to cleavage under reductive conditions, e.g. a disulfide bond.

[0160] One embodiment of the first series of aspects and embodiments of the present invention is a scaffold according to the present invention, wherein at least one biologically active molecule is covalently attached to the polymeric or oligomeric structure via a cleavable bond, said cleavable bond undergoing cleavage in vivo under acidic conditions present in the endosomes and / or lysosomes of mammalian cells, preferably human cells, preferably at a pH between 4.0 and 6.5, more preferably at a pH ≦ 5.5.

[0161] One embodiment of the first series of aspects and embodiments of the present invention is a scaffold according to the present invention, wherein at least one biologically active molecule is covalently attached to the polymeric or oligomeric structure of the scaffold via an imine bond, a hydrazone bond, a hydrazide bond, an oxime bond, a 1,3-dioxolane bond, a disulfide bond, a thio-ether bond, an amide bond, a peptide bond, or an ester bond, preferably via at least one linker.

[0162] One embodiment of the first series of aspects and embodiments of the present invention is a scaffold according to the present invention, wherein an aldehyde functional group at position C-23 of at least one saponin is involved in covalent attachment to the polymeric or oligomeric structure of the scaffold, and / or, if present, a glucuronic acid functional group on a carbohydrate substituent at the C-3 beta-OH group of at least one saponin is involved in covalent attachment to the polymeric or oligomeric structure of the scaffold, either via a direct bond or via at least one linker.

[0163] One embodiment of the first series of aspects and embodiments of the invention is a scaffold according to the invention, wherein the aldehyde functionality at position C-23 of at least one saponin is covalently coupled to a linker N-ε-maleimidocaproic acid hydrazide, which is covalently coupled via a thio-ether bond to a sulfhydryl group, such as a sulfhydryl group of a cysteine, on the polymeric or oligomeric structure of the scaffold.

[0164] One embodiment of the first series of aspects and embodiments of the invention is a scaffold according to the invention, wherein a glucuronic acid functionality on a carbohydrate substituent at the C-3 beta-OH group of at least one saponin is covalently coupled to a linker, 1-[bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium 3-oxide hexafluorophosphate, which is covalently coupled via an amide bond to an amine group on the polymeric or oligomeric structure of the scaffold, for example a lysine or N-terminal amine group of a proteinaceous molecule.

[0165] An embodiment of the first series of aspects and embodiments of the present invention is a scaffold according to the present invention, wherein the chemical group for covalent coupling of the scaffold to a carrier molecule is a click chemistry group.

[0166] An embodiment of the first series of aspects and embodiments of the present invention is a scaffold according to the present invention, wherein the click chemistry group is a tetrazine, azide, alkene, or alkyne, or a cyclic derivative of any of these groups, preferably an azide.

[0167] An embodiment of the first series of aspects and embodiments of the present invention is a scaffold according to the present invention, wherein the scaffold is a trifunctional linker and comprises a second chemical group to which at least one biologically active molecule is covalently attached, comprises a third chemical group for covalent attachment to a molecule, and comprises a first chemical group for covalent attachment to a carrier, preferably the trifunctional linker is of Scheme II and Structure B.

[0168] One embodiment of the first series of aspects and embodiments of the invention is a scaffold according to the invention, wherein at least one biologically active molecule is a defined number or range of glycoside molecules, preferably 1 to 128 or at least 2, 3, 4, 5, 6, 8, 10, 16, 32, 64, or 128 glycoside molecules, or any number therebetween, e.g., 7, 9, 12 glycoside molecules.

[0169] An embodiment of the first series of aspects and embodiments of the present invention is a scaffold according to the present invention, wherein the polymeric or oligomeric structure comprises a linear, branched, and / or cyclic polymer, oligomer, dendrimer, dendron, dendronized polymer, dendronized oligomer, DNA, polypeptide, polylysine, polyethylene glycol, or an assembly of these polymeric or oligomeric structures, which assembly is preferably assembled by covalent crosslinking.

[0170] An embodiment of the first series of aspects and embodiments of the invention is a scaffold according to the invention, wherein the carrier molecule comprises or consists of a covalent conjugate or complex of any of a proteinaceous molecule, a protein, a peptide, a nucleic acid, an oligonucleotide, a lipid, a fat, a fatty acid, a nanoparticle, a carbohydrate, or a combination thereof.

[0171] An embodiment of the first series of aspects and embodiments of the invention is a scaffold according to the invention, wherein the carrier molecule comprises or consists of an immunoglobulin, at least one binding domain of an immunoglobulin, and / or at least one binding fragment of an immunoglobulin, such as an antibody, an IgG, a molecule comprising or consisting of a Vhh domain or a Vh domain, a Fab, scFv, Fv, dAb, F(ab)2, Fcab fragment, or comprises or consists of at least one non-proteinaceous ligand and / or at least one proteinaceous ligand for binding to a cell surface molecule such as EGF or a cytokine.

[0172] One embodiment of the first series of aspects and embodiments of the invention is a scaffold according to the invention, wherein the carrier molecule is selected from the group consisting of CD71, CA125, EpCAM (17-1A), CD52, CEA, CD44v6, FAP, EGF-IR, integrin, syndecan-1, vascular integrin alpha-V beta-3, HER2, EGFR, CD20, CD22, folate receptor 1, CD146, CD56, CD19, CD138, CD27L receptor, PSMA, CanAg, integrin-alpha V, CA6, CD33, mesothelin, Cripto, CD3, CD30, CD239, CD70, CD1 23, CD352, DLL3, CD25, ephrinA4, MUC1, Trop2, CEACAM5, CEACAM6, HER3, CD74, PTK7, Notch3, FGF2, C4.4A, FLT3, CD38, FGFR3, CD7, PD-L1, CTLA4, CD52, PDGFRA, VEGFR1, VEGFR2, preferably CD71, EGFR, HER2.

[0173] An embodiment of the first series of aspects and embodiments of the invention is a scaffold according to the invention, wherein the carrier molecule comprises or consists of any one of cetuximab, daratumumab, gemtuzumab, trastuzumab, panitumumab, brentuximab, inotuzumab, moxetumomab, polatuzumab, obinutuzumab, OKT-9 anti-CD71 monoclonal antibody of the IgG type, pertuzumab, rituximab, ofatumumab, herceptin, alemtuzumab, pinatuzumab, OKT-10 anti-CD38 monoclonal antibody, an antibody of Table A2 or Table A3 or Table A4, preferably cetuximab or trastuzumab or OKT-9, or at least one tumor cell receptor binding fragment and / or at least one tumor cell receptor binding domain thereof, such as at least one tumor cell specific receptor binding fragment and / or at least one tumor cell specific receptor binding domain thereof.

[0174] An embodiment of the first series of aspects and embodiments of the present invention is a scaffold according to the present invention, wherein the scaffold is suitable for forming a covalent bond with a carrier molecule, said covalent bond preferably involving a cysteine ​​side chain of the carrier molecule and / or a lysine side chain of the carrier molecule, wherein the carrier molecule comprises at least cysteine ​​and / or lysine.

[0175] An embodiment of the first series of aspects and embodiments of the invention is a scaffold according to the invention, wherein the carrier molecule comprises or consists of at least one effector molecule, or the carrier further comprises at least one effector molecule, which is at least one of a drug substance, such as a payload, a toxin, a drug, a polypeptide, an oligonucleotide, a nucleic acid, a xenonucleic acid, an enzyme, such as urease and Cre recombinase, a protein toxin, or a ribosome-inactivating protein.

[0176] An embodiment of the first series of aspects and embodiments of the present invention is a scaffold according to the present invention, wherein the scaffold according to the present invention is a protein toxin selected from Table A5, and / or a viral toxin, such as apoptin; a bacterial toxin, such as Shiga toxin, Shiga toxin-like toxin, Pseudomonas aeruginosa exotoxin (PE), or exotoxin A of PE, full-length or truncated diphtheria toxin (DT), cholera toxin; fungal toxins, such as alpha-sarcin; ribosome-inactivating proteins, and plant toxins including the A chain of type 2 ribosome-inactivating proteins, such as giantin, e.g., giantin-30 or giantin-32, saporin, e.g., saporin-S3 or saporin-S6, bouganin or a deimmunized derivative of bouganin, debouganin, Shiga-like toxin A, pokeweed antiviral protein, ricin, ricin A chain, modeccin, modeccin A chain, abrin, abrin A chain, volkensin, volkensin A chain, viscumin, viscumin A chain; or animal or human toxins, such as frog RNase, or granzyme B, or angiogenin from humans, or any fragment or derivative thereof; preferably, the protein toxin is giantin and / or saporin.

[0177] In one embodiment of the first series of aspects and embodiments of the invention, the scaffold according to the invention is a vector, a gene, a transgene that induces cell suicide, deoxyribonucleic acid (DNA), ribonucleic acid (RNA), antisense oligonucleotide (ASO, AON), short interfering RNA (siRNA), microRNA (miRNA), DNA aptamer, RNA aptamer, mRNA, minicircle DNA, peptide nucleic acid (PNA), phosphoramidate morpholino oligomer. The nucleic acid may comprise or consist of one or more of: (PMO), locked nucleic acid (LNA), bridged nucleic acid (BNA), 2'-deoxy-2'-fluoroarabinonucleic acid (FANA), 2'-O-methoxyethyl-RNA (MOE), 2'-O,4'-aminoethylene-bridged nucleic acid, 3'-fluorohexitol nucleic acid (FHNA), plasmid, glycol nucleic acid (GNA), and threose nucleic acid (TNA), or derivatives thereof, preferably BNA, for example, BNA for silencing HSP27 protein expression.

[0178] An embodiment of the first series of aspects and embodiments of the present invention is a scaffold according to the present invention, wherein the effector molecule comprises or consists of at least one payload, preferably any one or more of a ribosome-targeting toxin, an elongation factor-targeting toxin, a tubulin-targeting toxin, a DNA-targeting toxin, and an RNA-targeting toxin, more preferably emtansine, pasudotox, maytansinoid derivative DM1, maytansinoid derivative DM4, monomethyl auristatin E (MMAE, vedotin), monomethyl auristatin F (MMAF, mafodotin), caliche Amicin, N-acetyl-γ-calicheamicin, pyrrolobenzodiazepine (PBD) dimer, benzodiazepine, CC-1065 analog, duocarmycin, doxorubicin, paclitaxel, cisplatin, cyclophosphamide, etoposide, docetaxel, 5-fluorouracil (5-FU), mitoxantrone, tubulysin, indolinobenzodiazepine, AZ13599185, cryptophycin, rhizoxin, methotrexate, anthracycline, camptothecin analog, SN-38, DX-8951f, exatecan mesylate, Pseudomonas The steroid inhibitor is selected from one or more of a truncated form of P. aeruginosa exotoxin (PE38), a duocarmycin derivative, amanitin, α-amanitin, splicostatin, tylanstatin, ozogamicin, tesirin, amberstatin 269, and soravtansine, or a derivative thereof.

[0179] An embodiment of the first series of aspects and embodiments of the invention is a scaffold according to the invention, wherein the carrier molecule comprises or consists of a covalently linked combination of an effector molecule and a monoclonal antibody, preferably selected from gemtuzumab ozogamicin, brentuximab vedotin, trastuzumab emtansine, inotuzumab ozogamicin, moxetumomab pasudotox, and polatuzumab vedotin, and the antibody-drug conjugates of Table A2 and Table A3.

[0180] An embodiment of the first series of aspects and embodiments of the present invention relates to a method for producing a scaffold suitable for covalently linking at least one biologically active molecule to a carrier molecule, the method comprising: a) providing a polymeric or oligomeric structure comprising a first chemical group for covalently coupling the polymeric or oligomeric structure to the carrier molecule and comprising at least one second chemical group different from the first chemical group, each second chemical group being for covalently coupling one of the at least one biologically active molecule to the oligomeric or polymeric structure; and b) covalently coupling at least one biologically active molecule to said polymeric or oligomeric structure via the second chemical group(s), preferably wherein the biologically active molecule(s) is / are any one of the biologically active molecules of the present invention, more preferably SO1861 and / or GE1741 and / or SA1641 and / or QS-21, thereby providing a scaffold.

[0181] An embodiment of the first series of aspects and embodiments of the present invention relates to a method according to the present invention, wherein a scaffold comprises at least one covalently attached biologically active molecule, the method comprising: a) providing a scaffold comprising at least one biologically active molecule covalently attached to the polymeric or oligomeric structure of said scaffold, preferably a scaffold according to the present invention or a scaffold obtainable by a method of the present invention or a scaffold obtained by a method of the present invention; and b) covalently coupling the scaffold of a) to a carrier molecule according to the present invention, thereby providing a scaffold covalently attached to the carrier molecule, wherein the scaffold comprises at least one covalently attached biologically active molecule.

[0182] One embodiment of the first series of aspects and embodiments of the invention is a method according to the invention, wherein a scaffold is capable of increasing endosomal escape and / or lysosomal escape of an effector molecule according to the invention, wherein said effector molecule is either covalently attached to the scaffold and contacted with a mammalian cell, or wherein said effector molecule is contacted with a mammalian cell in the presence of the scaffold.

[0183] An aspect of the invention of the second series of aspects and embodiments of the invention relates to a first proteinaceous molecule comprising a first binding site for binding to a first epitope of a first cell surface molecule, wherein the first proteinaceous molecule is provided with at least one saponin covalently attached to an amino acid residue of said first proteinaceous molecule via at least one linker and / or via an oligomeric or polymeric backbone or directly covalently attached to an amino acid residue of said first proteinaceous molecule.

[0184] An embodiment of the second series of aspects and embodiments of the invention is a first proteinaceous molecule according to the invention, wherein the first binding site comprises or consists of an immunoglobulin, or at least one binding domain of an immunoglobulin, and / or at least one binding fragment of an immunoglobulin, such as an antibody, an IgG, a molecule comprising or consisting of a Vhh domain or a Vh domain, a Fab, scFv, Fv, dAb, F(ab)2, Fcab fragment, and / or comprises or consists of at least one ligand for binding to a cell surface molecule such as EGF or a cytokine.

[0185] One embodiment of the second series of aspects and embodiments of the present invention is a first proteinaceous molecule according to the present invention, wherein the first epitope of the first cell surface molecule is a tumor cell-specific first epitope of a first tumor cell surface molecule, more preferably a tumor cell-specific first epitope of a first tumor cell surface receptor that is specifically present on tumor cells.

[0186] An embodiment of the second series of aspects and embodiments of the present invention is a first proteinaceous molecule according to the present invention, wherein at least one saponin is a triterpenoid saponin and / or a bisdesmosidic triterpene saponin belonging to the 12,13-dehydrooleanane type having an aldehyde functional group at position C-23 and optionally containing a glucuronic acid functional group in the carbohydrate substituent of the C-3 beta-OH group of the saponin, and / or a saponin isolated from Gypsophila species and / or Saponaria species and / or Agrostemma species and / or Quillaja species, e.g. Quillaja saponaria.

[0187] An embodiment of the second series of aspects and embodiments of the invention is a first proteinaceous molecule according to the invention, wherein the at least one saponin is a single specific saponin or a mixture of two or more different saponins, such as one or more of the saponins of Table A1 or Scheme I, SO1861, SA1657, GE1741, SA1641, QS-21, QS-21A, QS-21 A-api, QS-21 A-xyl, QS-21B, QS-21 B-api, QS-21 B-xyl, QS-7-xyl, QS-7-api, QS-17-api, QS-17-xyl, QS1861, QS1862, Quillaja saponin, Saponinum albumenum album, QS-18, Quil-A, Gyp1, gypsoside A, AG1, AG2, SO1542, SO1584, SO1658, SO1674, SO1832, or any stereoisomer thereof and / or any combination thereof. Preferably, the saponin is SO1861 and / or GE1741 and / or SA1641 and / or QS-21, and / or a saponin having a quillaic acid aglycone core, a Gal-(1→2)-[Xyl-(1→3)]-GlcA carbohydrate substituent at the C-3 beta-OH group, and a Glc-(1→3)-Xyl-(1→4)-Rha-(1→2)-[Xyl-(1→3)-4-OAc-Qui-(1→4)]-Fuc carbohydrate substituent at the C-28-OH group. and / or 3-O-beta-D-galactopyranosyl-(1→2)-[beta-D-xylopyranosyl-(1→3)]-beta-D-glucuronopyranosylquilayic acid, 28-O-beta-D-glucopyranosyl-(1→3)-beta-D-xylopyranosyl-(1→4)-alpha-L-rhamnopyranosyl-(1→2)-[beta-D-xylopyranosyl-(1→3)-4-OAc-beta-D-quinovopyranosyl-(1→4)]-beta-D-fucopyranoside. More preferably, the saponin is SO1861 and / or QS-21.

[0188] An embodiment of the second series of aspects and embodiments of the present invention is a first proteinaceous molecule according to the present invention, wherein at least one saponin is a bisdesmosidic saponin and has a molecular mass of at least 1,500 daltons and comprises an oleanane-type triterpene containing an aldehyde group at the C-23 position and optionally a hydroxyl group at the C-16 position, and has a first branched carbohydrate side chain at the C-3 position, which first branched carbohydrate side chain optionally contains glucuronic acid. The saponin contains an ester group with a second branched carbohydrate side chain at the C-28 position, this second branched carbohydrate chain preferably containing at least four carbohydrate units, optionally containing at least one acetyl residue, for example two acetyl residues, and / or optionally containing a deoxy carbohydrate, and / or optionally containing quinovose, and / or optionally containing glucose, and / or optionally containing 4-methoxycinnamic acid, and / or optionally containing 5-O-[5-O-Ara / Api-3,5-dihydroxy-6-methyl-octanoyl]-3,5-dihydroxy-6-methyl-octanoic acid, and / or optionally containing 5-O-[5-O-Rha-(1→2)-Ara / Api-3,5-dihydroxy-6-methyl-octanoyl]-3,5-dihydroxy-6-methyl-octanoic acid, linked to the carbohydrate via an ester bond. Alternatively, the at least one saponin is QS-21 or any one or more of QS-21A, QS-21 A-api, QS-21 A-xyl, QS-21B, QS-21 B-api, QS-21 B-xyl, QS-7-xyl, QS-7-api, QS-17-api, QS-17-xyl, QS-18, QS-1861, protonated QS1861 (QS1862), and Quil-A.

[0189] An embodiment of the second series of aspects and embodiments of the present invention is a first proteinaceous molecule according to the present invention, wherein at least one saponin is a bisdesmosidic triterpene saponin belonging to the type of 12,13-dehydrooleanane having an aldehyde functional group at position C-23, and wherein the at least one saponin is covalently coupled via the aldehyde functional group on the saponin, preferably said aldehyde functional group at position C-23, to an amino acid residue of the first proteinaceous molecule, preferably via at least one linker, preferably at least one linker, more preferably at least one cleavable linker, wherein the amino acid residue is preferably selected from cysteine ​​and lysine.

[0190] One embodiment is a first proteinaceous molecule according to the present invention, wherein at least one saponin is a bisdesmosidic triterpene saponin belonging to the 12,13-dehydrooleanane type having an aldehyde functional group at position C-23 and comprising a glucuronic acid functional group on the carbohydrate substituent at the C-3 beta-OH group of the saponin, and the at least one saponin is covalently coupled to an amino acid residue of the first proteinaceous molecule via the glucuronic acid functional group on the carbohydrate substituent at the C-3 beta-OH group of the saponin, preferably via at least one linker, the amino acid residue being preferably selected from cysteine ​​and lysine.

[0191] An embodiment of the second series of aspects and embodiments of the present invention is a first proteinaceous molecule according to the present invention, wherein the aldehyde functional group at position C-23 of at least one saponin is covalently bound to a linker N-ε-maleimidocaproic acid hydrazide, which linker is covalently bound to a sulfhydryl group, such as a sulfhydryl group of a cysteine, in the first proteinaceous molecule via a thio-ether bond.

[0192] An embodiment of the second series of aspects and embodiments of the present invention is a first proteinaceous molecule according to the present invention, wherein a glucuronic acid functional group in the carbohydrate substituent at the C-3 beta-OH group of at least one saponin is covalently attached to the linker 1-[bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium 3-oxide hexafluorophosphate, which linker is covalently attached via an amide bond to an amine group in the first proteinaceous molecule, for example an amine group of a lysine or N-terminal lysine of the first proteinaceous molecule.

[0193] An embodiment of the second series of aspects and embodiments of the invention is a first proteinaceous molecule according to the invention, wherein the first epitope of the first cell surface molecule to which the first binding site of the first proteinaceous molecule binds is preferably CD71, CA125, EpCAM (17-1A), CD52, CEA, CD44v6, FAP, EGF-IR, integrin, syndecan-1, vascular integrin alpha-V beta-3, HER2, EGFR, CD20, CD22, folate receptor 1, CD146, CD56, CD19, CD138, CD27L receptor, PSMA, CanAg, integrin-α, α-β, α-β, β ... The first tumor cell-specific epitope of a tumor cell-specific receptor selected from IL-1, IL-1, IL-2, IL-3, IL-4, IL-5, IL-6, IL-8, IL-1, IL-1, IL-2, IL-3, IL-4, IL-5, IL-6, IL-1, IL-2, IL-1, IL-2, IL-3, IL-4 ...

[0194] An embodiment of the second series of aspects and embodiments of the present invention is a first proteinaceous molecule according to the present invention, wherein the tumor cell-specific first epitope, first tumor cell surface molecule or first tumor cell-specific receptor is a first epitope or first molecule or first receptor that is internalized by a tumor cell after binding of the first proteinaceous molecule of the present invention to the first epitope, first molecule or first receptor, and preferably the first proteinaceous molecule, when bound to a cell surface molecule comprising the first epitope, tumor cell surface molecule or tumor cell-specific receptor, is subject to internalization mediated by the tumor cell receptor, e.g. via endocytosis, or internalization mediated by the tumor cell surface molecule, e.g. via endocytosis.

[0195] An embodiment of the second series of aspects and embodiments of the invention is a first proteinaceous molecule according to the invention, wherein the first binding site of the first proteinaceous molecule is selected from the group consisting of cetuximab, daratumumab, gemtuzumab, trastumumab, panitumumab, brentuximab, inotuzumab, moxetumomab, polatuzumab, obinutuzumab, OKT-9 anti-CD71 monoclonal antibody of IgG type, pertuzumab, rituximab, ofatumumab, herceptin, alemtuzumab, pinatuzumab , OKT-10 anti-CD38 monoclonal antibody, an antibody of Table A2 or Table A3 or Table A4, preferably cetuximab or trastuzumab or OKT-9, or at least one tumor cell receptor binding fragment and / or at least one tumor cell receptor binding domain thereof, preferably at least one tumor cell-specific receptor binding fragment and / or at least one tumor cell-specific receptor binding domain thereof.

[0196] One aspect of the invention of the second series of aspects and embodiments of the invention relates to a combination of therapeutic agents, the combination of therapeutic agents comprising: (a) a first pharmaceutical composition comprising a first proteinaceous molecule according to the invention, and optionally a pharmaceutically acceptable excipient; and (b) a second pharmaceutical composition comprising a second proteinaceous molecule different from the first proteinaceous molecule, wherein the second proteinaceous molecule comprises a second binding site for binding to a second epitope of the second cell surface molecule different from the first cell surface molecule and comprises an effector moiety, wherein the second pharmaceutical composition optionally further comprises a pharmaceutically acceptable excipient, and wherein the second epitope is different from the first epitope.

[0197] One embodiment of the second series of aspects and embodiments of the present invention is a therapeutic combination in accordance with the present invention, the therapeutic combination comprising: (a) a first pharmaceutical composition in accordance with the present invention, comprising a first proteinaceous molecule in accordance with the present invention, wherein the first epitope on the first cell surface molecule is a first tumor cell-specific epitope on a first tumor cell-specific surface molecule, preferably a first tumor cell-specific epitope on a first cell surface receptor that is specifically present on tumor cells; and (b) a second pharmaceutical composition in accordance with the present invention, wherein the second cell surface molecule is a second tumor cell-specific surface molecule different from the first tumor cell-specific surface molecule, preferably a second cell surface receptor that is specifically present on tumor cells different from the first cell surface receptor that is specifically present on tumor cells. The second epitope is a second tumor cell-specific epitope.

[0198] One embodiment of the second series of aspects and embodiments of the present invention is a therapeutic combination in accordance with the present invention, comprising: (a) a first pharmaceutical composition comprising a first proteinaceous molecule in accordance with the present invention and comprising a first binding site for binding to a first epitope on a first cell surface molecule, the first pharmaceutical composition optionally further comprising a pharmaceutically acceptable excipient; and a third pharmaceutical composition comprising a third proteinaceous molecule, the third proteinaceous molecule comprising a first binding site for binding to a first epitope on a cell surface molecule of (a) and an effector moiety, the third pharmaceutical composition optionally further comprising a pharmaceutically acceptable excipient. The first binding site of the first proteinaceous molecule and the first binding site of the third proteinaceous molecule are the same. The first cell surface molecule and the first epitope on the first cell surface molecule to which the first proteinaceous molecule can bind are the same as the first cell surface molecule and the first epitope on the first cell surface molecule to which the third proteinaceous molecule can bind.

[0199] One embodiment of the second series of aspects and embodiments of the present invention is a therapeutic combination according to the present invention, the therapeutic combination comprising (a) a first pharmaceutical composition according to the present invention and (b) a third pharmaceutical composition according to the present invention, wherein the first cell surface molecule is expressed on the surface of a tumor cell, preferably the first cell surface molecule is a tumor cell-specific surface molecule, and preferably the first epitope is a first tumor cell-specific epitope.

[0200] An embodiment of the second series of aspects and embodiments of the invention is a first proteinaceous molecule according to the invention, or a combination of therapeutic agents according to the invention, wherein the first binding site for binding to a first epitope on a first cell surface molecule is a binding site for a tumor cell-specific first epitope on a first cell surface receptor that is specifically present on tumor cells.

[0201] An embodiment of the second series of aspects and embodiments of the invention is a therapeutic combination according to the invention, wherein the second binding site of the second proteinaceous molecule and / or the first binding site of the third proteinaceous molecule comprises or consists of an immunoglobulin, at least one binding domain of an immunoglobulin, and / or at least one binding fragment of an immunoglobulin, such as an antibody, an IgG, a molecule comprising or consisting of a Vhh domain or a Vh domain, a Fab, scFv, Fv, dAb, F(ab)2, Fcab fragment, and / or comprises or consists of at least one ligand for binding to a cell surface molecule such as EGF or a cytokine.

[0202] An embodiment of the second series of aspects and embodiments of the invention is a therapeutic combination according to the invention, wherein the second binding site of the second proteinaceous molecule for binding to a second epitope is a second binding site for a tumor cell-specific second epitope on a second cell surface receptor that is specifically present on tumor cells, and the second binding site is different from the first binding site.

[0203] One embodiment of the second series of aspects and embodiments of the invention is a first proteinaceous molecule according to the invention, or a combination of therapeutic agents according to the invention, wherein said first and second proteinaceous molecules comprise first and second binding sites for binding to first and second tumor cell-specific epitopes on first and second tumor cell-specific receptors, respectively. The receptors are different and present on the same tumor cell. The first and second binding sites are different, and the first and second tumor cell-specific epitopes are different.

[0204] An embodiment of the second series of aspects and embodiments of the present invention is a first proteinaceous molecule according to the present invention, or a combination of therapeutic agents according to the present invention, wherein said first and third proteinaceous molecules comprise the same first binding site for binding to a first tumor cell-specific epitope on a first tumor cell-specific receptor.

[0205] An embodiment of the second series of aspects and embodiments of the invention is a first proteinaceous molecule according to the invention, or a combination of therapeutic agents according to the invention, wherein the first receptor and / or the second receptor is selected from the group consisting of CD71, CA125, EpCAM (17-1A), CD52, CEA, CD44v6, FAP, EGF-IR, integrin, syndecan-1, vascular integrin alpha-V beta-3, HER2, EGFR, CD20, CD22, folate receptor 1, CD146, CD56, CD19, CD138, CD27L receptor, PSMA, CanA g, integrin-alpha V, CA6, CD33, mesothelin, Cripto, CD3, CD30, CD239, CD70, CD123, CD352, DLL3, CD25, ephrin A4, MUC1, Trop2, CEACAM5, CEACAM6, HER3, CD74, PTK7, Notch3, FGF2, C4.4A, FLT3, CD38, FGFR3, CD7, PD-L1, CTLA4, CD52, PDGFRA, VEGFR1, VEGFR2, and preferably selected from CD71, EGFR, and HER2.

[0206] An embodiment of the second series of aspects and embodiments of the present invention is a first proteinaceous molecule according to the invention and / or a combination of therapeutic agents according to the invention, wherein the first and second tumor cell-specific receptors are internalized by the tumor cell after binding to the first proteinaceous molecule according to the invention and / or the second proteinaceous molecule according to the invention. Preferably, binding of the first proteinaceous molecule and / or the second proteinaceous molecule to the first and second tumor cell-specific receptors, respectively, results in tumor cell receptor-mediated internalization of the complex of the first proteinaceous molecule with the first tumor cell-specific receptor and the complex of the second proteinaceous molecule with the second tumor cell-specific receptor, e.g. via endocytosis.

[0207] An embodiment of the second series of aspects and embodiments of the present invention is a therapeutic combination or a first pharmaceutical composition according to the present invention, wherein the first tumor cell receptor, preferably a first tumor cell-specific receptor, is internalized by the tumor cell after binding to the first proteinaceous molecule according to the present invention and / or after binding to the third proteinaceous molecule according to the present invention. Preferably, binding of the first proteinaceous molecule and / or the third proteinaceous molecule to the first tumor cell receptor, e.g., the first tumor cell-specific receptor, is followed by tumor cell receptor-mediated internalization of the complex of the first proteinaceous molecule with the first tumor cell receptor and the complex of the third proteinaceous molecule with the first tumor cell receptor, e.g., via endocytosis.

[0208] An embodiment of the second series of aspects and embodiments of the present invention is a first proteinaceous molecule according to the invention and / or a therapeutic combination according to the invention, wherein the first binding site and / or the second binding site is or comprises a monoclonal antibody, or at least one cell surface molecule binding fragment and / or domain thereof, preferably selected from the group consisting of cetuximab, daratumumab, gemtuzumab, trastuzumab, panitumumab, brentuximab, inotuzumab, and any one of the antibodies of Table A4, preferably cetuximab or trastuzumab or OKT-9, or at least one cell surface molecule-binding fragment or domain thereof, with the proviso that the first binding site of the first proteinaceous molecule is different from the second binding site of the second proteinaceous molecule.

[0209] An embodiment of the second series of aspects and embodiments of the present invention is a therapeutic combination according to the present invention, or a first pharmaceutical composition according to the present invention, wherein the first binding site of the first proteinaceous molecule and the third proteinaceous molecule comprises at least one monoclonal antibody or cell surface molecule binding domain and / or fragment thereof, preferably cetuximab, daratumumab, gemtuzumab, trastumumab, panitumumab, brentuximab, inotuzumab, moxetumomab, tuzumab, polatuzumab, obinutuzumab, OKT-9 anti-CD71 monoclonal antibody of the IgG type, pertuzumab, rituximab, ofatumumab, herceptin, alemtuzumab, pinatuzumab, OKT-10 anti-CD38 monoclonal antibody, an antibody of Table A2 or Table A3 or Table A4, preferably cetuximab or trastuzumab or OKT-9, or at least one cell surface molecule binding fragment and / or domain thereof, with the proviso that the first binding site of the first proteinaceous molecule is the same as the first binding site of the third proteinaceous molecule.

[0210] An embodiment of the second set of aspects and embodiments of the invention is a therapeutic combination according to the invention, wherein the second binding site of the second proteinaceous molecule and / or the first binding site of the third proteinaceous molecule is or comprises a monoclonal antibody or at least one cell surface molecule-binding fragment or domain thereof, and preferably comprises or consists of gemtuzumab ozogamicin, brentuximab vedotin, trastuzumab emtansine, inotuzumab ozogamicin, moxetumomab pasudotox, and polatuzumab vedotin, and any one of the antibody-drug conjugates of Table A2 and Table A3.

[0211] An embodiment of the second series of aspects and embodiments of the invention is a therapeutic combination according to the invention, wherein the effector moiety comprised by the second proteinaceous molecule and / or the third proteinaceous molecule comprises or consists of one or more of an oligonucleotide, a nucleic acid, a xenonucleic acid, preferably a vector, a gene, a transgene inducing cell suicide, deoxyribonucleic acid (DNA), ribonucleic acid (RNA), an antisense oligonucleotide (ASO, AON), a short interfering RNA (siRNA), a microRNA (miRNA), a DNA aptamer, an RNA aptamer, an mRNA, a mini-RNA, a ribonucleic acid (Xenonucleic acid), a vector, a gene, a transgene inducing cell suicide, a deoxyribonucleic acid (DNA), a ribonucleic acid (RNA), an antisense oligonucleotide (ASO, AON), a small interfering RNA (siRNA), a microRNA (miRNA), a DNA aptamer, an RNA aptamer, an mRNA, a mini-RNA ... The nucleic acid may be selected from any one or more of clone DNA, peptide nucleic acid (PNA), phosphoramidate morpholino oligomer (PMO), locked nucleic acid (LNA), bridged nucleic acid (BNA), 2'-deoxy-2'-fluoroarabinonucleic acid (FANA), 2'-O-methoxyethyl-RNA (MOE), 2'-O,4'-aminoethylene-bridged nucleic acid, 3'-fluorohexitol nucleic acid (FHNA), plasmid, glycol nucleic acid (GNA), and threose nucleic acid (TNA), or derivatives thereof, more preferably BNA, for example, BNA for silencing HSP27 protein expression.

[0212] An embodiment of the second series of aspects and embodiments of the invention is a therapeutic combination according to the invention, wherein the effector moiety comprised by the second proteinaceous molecule and / or the third proteinaceous molecule comprises or consists of at least one proteinaceous molecule, preferably selected from any one or more of peptides, proteins, enzymes such as urease and Cre recombinase, ribosome-inactivating proteins, proteinaceous toxins, more preferably proteinaceous toxins selected from Table A5, and / or viral toxins, such as apoptin; bacterial toxins, such as Shiga toxins, Shiga-like toxins, Pseudomonas The protein toxin is selected from any one or more of: P. aeruginosa exotoxin (PE), or exotoxin A of PE, full-length or truncated diphtheria toxin (DT), cholera toxin; fungal toxins, such as alpha-sarcin; plant toxins, including ribosome-inactivating proteins and the A chain of type 2 ribosome-inactivating proteins, such as giantin, e.g., giantin-30 or giantin-32, saporin, e.g., saporin-S3 or saporin-S6, bouganin or a deimmunized derivative of bouganin, debouganin, Shiga-like toxin A, pokeweed antiviral protein, ricin, ricin A chain, modeccin, modeccin A chain, abrin, abrin A chain, volkensin, volkensin A chain, viscumin, viscumin A chain; or animal or human toxins, such as frog RNase, granzyme B, or angiogenin from humans, or any fragment or derivative thereof. Preferably, the protein toxin is giantin and / or saporin.

[0213] An embodiment of the second series of aspects and embodiments of the invention is a therapeutic combination according to the invention, wherein the effector moiety comprised by the second proteinaceous molecule and / or the third proteinaceous molecule comprises or consists of at least one payload, preferably any one or more of a ribosome-targeting toxin, an elongation factor-targeting toxin, a tubulin-targeting toxin, a DNA-targeting toxin, and an RNA-targeting toxin, more preferably emtansine, passudotox, maytansinoid derivative DM1, maytansinoid derivative DM4, monomethyl auristatin E (MMAE, vedotin), monomethyl auristatin B (MMAE, vedotin), or a toxin that targets ribosomes, elongation factors, tubulin, DNA, or RNA, more preferably emtansine, passudotox, maytansinoid derivative DM1, maytansinoid derivative DM4, monomethyl auristatin C (MMAE, vedotin), monomethyl auristatin D (MMAE, vedotin), or a toxin that targets RNA. methicone F (MMAF, mafodotin), calicheamicin, N-acetyl-γ-calicheamicin, pyrrolobenzodiazepine (PBD) dimer, benzodiazepine, CC-1065 analog, duocarmycin, doxorubicin, paclitaxel, docetaxel, cisplatin, cyclophosphamide, etoposide, docetaxel, 5-fluorouracil (5-FU), mitoxantrone, tubulysin, indolinobenzodiazepine, AZ13599185, cryptophycin, rhizoxin, methotrexate, anthracyclines, camptothecin analog, SN-38, DX-8951f, exatecan mesylate, Pseudomonas The steroid inhibitor is selected from one or more of a truncated form of P. aeruginosa exotoxin (PE38), a duocarmycin derivative, amanitin, α-amanitin, splicostatin, tylanstatin, ozogamicin, tesirin, amberstatin 269, and soravtansine, or a derivative thereof.

[0214] An embodiment of the second set of aspects and embodiments of the present invention is a first proteinaceous molecule according to the present invention, wherein the first proteinaceous molecule comprises more than one saponin, preferably 2, 3, 4, 5, 6, 8, 10, 16, 32, 64, or 1-100 saponins, or any number of saponins in between, for example 7, 9, 12 saponins, covalently attached directly to an amino acid residue of the first proteinaceous molecule, preferably to a cysteine ​​and / or a lysine, and / or via at least one linker and / or via at least one cleavable linker, and / or to at least one polymeric or oligomeric backbone, preferably via 1 to 8 such backbones or 2 to 4 such backbones. At least one scaffold is optionally dendron-based, and 1 to 32 saponins, for example, 2, 3, 4, 5, 6, 8, 10, 16, 32 saponins, or any number therebetween, for example, 7, 9, 12 saponins, are covalently attached to the at least one scaffold.

[0215] An embodiment of the second series of aspects and embodiments of the invention is a first proteinaceous molecule according to the invention, wherein at least one linker is a non-cleavable linker or a cleavable linker. A cleavable linker is subject to cleavage, for example, under acidic, reductive, enzymatic or photoinduced conditions, preferably the cleavable linker comprises a hydrazone or hydrazide bond that is subject to cleavage under acidic conditions when bound to saponin, and / or comprises a bond that is susceptible to proteolysis, for example proteolysis by cathepsin B, when bound to saponin, and / or is a bond that is susceptible to cleavage under reductive conditions, for example a disulfide bond.

[0216] An embodiment of the second set of aspects and embodiments of the present invention is a first proteinaceous molecule according to the invention, wherein the cleavable linker undergoes cleavage in vivo under acidic conditions present in the endosomes and / or lysosomes of mammalian cells, preferably human cells, when the cleavable linker is attached to a saponin, preferably at a pH between 4.0 and 6.5, more preferably at a pH ≦ 5.5.

[0217] An embodiment of the second series of aspects and embodiments of the present invention is a first proteinaceous molecule according to the present invention, wherein the oligomeric or polymeric backbone comprises a polymeric or oligomeric structure and comprises chemical groups, chemical groups for covalent coupling of the backbone to amino acid residues of said first proteinaceous molecule.

[0218] An embodiment of the second set of aspects and embodiments of the present invention is a first proteinaceous molecule according to the present invention, wherein at least one saponin is covalently attached to the polymer or oligomeric structure of the oligomeric or polymer backbone via at least one cleavable linker according to the present invention.

[0219] An embodiment of the second series of aspects and embodiments of the present invention is a first proteinaceous molecule according to the present invention, wherein the chemical group of the oligomeric or polymeric backbone for covalent coupling of the oligomeric or polymeric backbone to an amino acid residue of said first proteinaceous molecule is a click chemistry group, preferably selected from tetrazine, azide, alkene or alkyne, or cyclic derivatives of these groups, more preferably said chemical group is azide.

[0220] An embodiment of the second series of aspects and embodiments of the present invention is a first proteinaceous molecule according to the present invention, wherein the polymer or oligomeric structure of the oligomeric or polymer backbone comprises a linear, branched, and / or cyclic polymer, oligomer, dendrimer, dendron, dendronized polymer, dendronized oligomer, DNA, polypeptide, polylysine, polyethylene glycol, or an assembly of these polymer or oligomeric structures, which assembly is preferably assembled by covalent crosslinking.

[0221] An aspect of the invention of the second series of aspects and embodiments of the invention relates to a composition, comprising a first proteinaceous molecule according to the invention and a second proteinaceous molecule according to the invention.

[0222] An aspect of the invention of the second series of aspects and embodiments of the invention relates to a composition, comprising a first proteinaceous molecule according to the invention and a third proteinaceous molecule according to the invention.

[0223] An embodiment of the second series of aspects and embodiments of the present invention is a composition according to the present invention, wherein the effector moiety comprised by the second proteinaceous molecule or the third proteinaceous molecule is any one of the effector moieties according to the present invention, preferably a BNA.

[0224] A second series of aspects and certain aspects of embodiments of the present invention relate to compositions comprising a first proteinaceous molecule according to the present invention and a target molecule, preferably a vector, a gene, a transgene inducing cell suicide, deoxyribonucleic acid (DNA), ribonucleic acid (RNA), antisense oligonucleotides (ASO, AON), short interfering RNA (siRNA), microRNA (miRNA), DNA aptamer, RNA aptamer, mRNA, minicircle DNA, peptide nucleic acid (PNA), phosphoramidate morpholino oligomer (PMO), locked nucleic acid (LN). A), an oligonucleotide, a nucleic acid, and a xenonucleic acid selected from at least one of bridged nucleic acids (BNA), 2'-deoxy-2'-fluoroarabinonucleic acid (FANA), 2'-O-methoxyethyl-RNA (MOE), 2'-O,4'-aminoethylene-bridged nucleic acid, 3'-fluorohexitol nucleic acid (FHNA), a plasmid, glycol nucleic acid (GNA), and threose nucleic acid (TNA), or derivatives thereof, more preferably BNA, for example, BNA for silencing HSP27 protein expression.

[0225] An aspect of the invention of the second series of aspects and embodiments of the invention relates to an antibody-drug conjugate or a ligand-drug conjugate, comprising a first proteinaceous molecule according to the invention and an effector moiety.

[0226] In one embodiment of the second series of aspects and embodiments of the invention is an antibody-drug conjugate or a ligand-drug conjugate according to the invention, wherein the antibody is selected from the group consisting of CD71, CA125, EpCAM (17-1A), CD52, CEA, CD44v6, FAP, EGF-IR, integrin, syndecan-1, vascular integrin alpha-V beta-3, HER2, EGFR, CD20, CD22, folate receptor 1, CD146, CD56, CD19, CD138, CD27L receptor, PSMA, CanAg, integrin. The antibody may bind to any one of: leukocyte-alpha V, CA6, CD33, mesothelin, Cripto, CD3, CD30, CD239, CD70, CD123, CD352, DLL3, CD25, ephrin A4, MUC1, Trop2, CEACAM5, CEACAM6, HER3, CD74, PTK7, Notch3, FGF2, C4.4A, FLT3, CD38, FGFR3, CD7, PD-L1, CTLA4, CD52, PDGFRA, VEGFR1, VEGFR2, preferably CD71, HER2, and EGFR. and / or is or comprises any one of cetuximab, daratumumab, gemtuzumab, trastuzumab, panitumumab, brentuximab, inotuzumab, moxetumomab, polatuzumab, obinutuzumab, OKT-9 anti-CD71 monoclonal antibody of the IgG type, pertuzumab, rituximab, ofatumumab, herceptin, alemtuzumab, pinatuzumab, OKT-10 anti-CD38 monoclonal antibody, an antibody of Table A2 or Table A3 or Table A4, preferably cetuximab or trastuzumab or OKT-9, or at least one tumor cell receptor-binding fragment thereof and / or at least one tumor cell receptor-binding domain thereof. and / or the antibody-drug conjugate comprises gemtuzumab ozogamicin, brentuximab vedotin, trastuzumab emtansine, inotuzumab ozogamicin, moxetumomab pasudotox, and polatuzumab vedotin, and any one of the antibody-drug conjugates of Tables A2 and A3. Alternatively, the ligand-drug conjugate comprises at least one ligand for binding to a cell surface molecule, such as EGF or a cytokine.

[0227] An embodiment of the second set of aspects and embodiments of the invention is an antibody-drug conjugate or a ligand-drug conjugate according to the invention, wherein the effector moiety is any one or more of the effector moieties according to the invention.

[0228] An aspect of the invention of the second series of aspects and embodiments of the invention relates to a pharmaceutical composition, comprising a composition according to the invention, or an antibody-drug conjugate according to the invention, or a ligand-drug conjugate according to the invention, and optionally further comprising a pharmaceutically acceptable excipient.

[0229] An aspect of the invention in the second series of aspects and embodiments of the invention relates to a therapeutic combination, or composition, or antibody-drug conjugate, or ligand-drug conjugate, or pharmaceutical composition according to the invention for use as a medicament.

[0230] An aspect of the invention in the second series of aspects and embodiments of the invention relates to a therapeutic combination, or composition, or antibody-drug conjugate, or ligand-drug conjugate, or pharmaceutical composition according to the invention for use in the treatment or prevention of cancer or autoimmune disease.

[0231] An aspect of the invention of the third series of aspects and embodiments of the invention relates to an effector moiety capable of inducing an intracellular effect when present in a mammalian cell, wherein the effector moiety is conjugated to at least one saponin, which is covalently attached to the effector moiety via at least one linker or directly covalently attached to said effector moiety.

[0232] An embodiment of the third series of aspects and embodiments of the invention is an effector moiety according to the invention, comprising or consisting of an oligonucleotide, a nucleic acid, a xenonucleic acid, preferably a vector, a gene, a transgene inducing cell suicide, deoxyribonucleic acid (DNA), ribonucleic acid (RNA), antisense oligonucleotides (ASO, AON), short interfering RNA (siRNA), microRNA (miRNA), a DNA aptamer, an RNA aptamer, mRNA, minicircle DNA, peptide nucleic acid (PNA), phosphoramidate, The nucleic acid may be selected from one or more of a morpholino oligomer (PMO), a locked nucleic acid (LNA), a bridged nucleic acid (BNA), a 2'-deoxy-2'-fluoroarabinonucleic acid (FANA), a 2'-O-methoxyethyl-RNA (MOE), a 2'-O,4'-aminoethylene-bridged nucleic acid, a 3'-fluorohexitol nucleic acid (FHNA), a plasmid, a glycol nucleic acid (GNA), and a threose nucleic acid (TNA), or a derivative thereof, more preferably a BNA, for example a BNA for silencing HSP27 protein expression.

[0233] An embodiment of the third series of aspects and embodiments of the invention is an effector moiety according to the invention comprising at least one proteinaceous molecule, preferably selected from a peptide, a protein, an enzyme such as urease and Cre recombinase, a ribosome-inactivating protein, a proteinaceous toxin, such as any one or more of the protein toxins selected from Table A5, and / or any one or more bacterial or plant toxins, more preferably a viral toxin, such as apoptin; a bacterial toxin, such as Shiga toxin, Shiga-like toxin, Pseudomonas The protein toxin is selected from any one or more of: P. aeruginosa exotoxin (PE), or exotoxin A of PE, full-length or truncated diphtheria toxin (DT), cholera toxin; fungal toxins, such as alpha-sarcin; plant toxins, including ribosome-inactivating proteins and type 2 ribosome-inactivating protein A chains, such as giantin-30 or giantin-32, saporin-S3 or saporin-S6, bouganin or the deimmunized derivative of bouganin, debouganin, Shiga-like toxin A, pokeweed antiviral protein, ricin, ricin A chain, modeccin, modeccin A chain, abrin, abrin A chain, volkensin, volkensin A chain, viscumin, viscumin A chain; or animal or human toxins, such as frog RNase, or granzyme B or angiogenin from humans, or any fragment or derivative thereof. Preferably, the protein toxin is giantin and / or saporin.

[0234] An embodiment of the third series of aspects and embodiments of the invention is an effector moiety according to the invention, comprising at least one payload, preferably any one or more of a ribosome-targeting toxin, an elongation factor-targeting toxin, a tubulin-targeting toxin, a DNA-targeting toxin, and an RNA-targeting toxin, more preferably emtansine, pasudotox, maytansinoid derivative DM1, maytansinoid derivative DM4, monomethyl auristatin E (MMAE, vedotin), monomethyl auristatin F (MMAF, mafodotin), calicheamicin, N-acetyl-γ-calicheamicin, pyrrolobenzodiazepine (PBD) dimer, benzodiazepine, CC-1065 analog, duocarmycin, doxorubicin, paclitaxel, docetaxel, cisplatin, cyclophosphamide, etoposide, docetaxel, 5-fluorouracil (5-FU), mitoxantrone, tubulysin, indolinobenzodiazepine, AZ13599185, cryptophycin, rhizoxin, methotrexate, anthracycline, camptothecin analog, SN-38, DX-8951f, exatecan mesylate, Pseudomonas The steroid inhibitor is selected from one or more of a truncated form of P. aeruginosa exotoxin (PE38), a duocarmycin derivative, amanitin, α-amanitin, splicostatin, tylanstatin, ozogamicin, tesirin, amberstatin 269, and soravtansine, or a derivative thereof.

[0235] An embodiment of the third series of aspects and embodiments of the present invention is an effector moiety according to the present invention, wherein at least one saponin is a triterpenoid saponin or a bisdesmosidic triterpene saponin belonging to the 12,13-dehydrooleanane type having an aldehyde functionality at position C-23 and optionally a glucuronic acid functionality in the carbohydrate substituent of the C-3 beta-OH group of the saponin, and / or a saponin isolated from Gypsophila species and / or Saponaria species and / or Agrostemma species and / or Quillaja species, e.g. Quillaja saponaria.

[0236] An embodiment of the third series of aspects and embodiments of the invention is an effector moiety according to the invention, wherein the at least one saponin is a single particular saponin or a mixture of two or more different saponins.

[0237]

[0023] An embodiment of the third series of aspects and embodiments of the invention is an effector moiety according to the invention, wherein the saponin is one or more of the saponins of Table A1 or Scheme I, SO1861, SA1657, GE1741, SA1641, QS-21, QS-21A, QS-21 A-api, QS-21 A-xyl, QS-21B, QS-21 B-api, QS-21 B-xyl, QS-7-xyl, QS-7-api, QS-17-api, QS-17-xyl, QS1861, QS1862, Quillaja saponin, Saponinum albumenum album, QS-18, Quil-A, Gyp1, gypsoside A, AG1, AG2, SO1542, SO1584, SO1658, SO1674, SO1832, or any stereoisomer thereof and / or any combination thereof. Preferably, the saponin is SO1861 and / or GE1741 and / or SA1641 and / or QS-21, and / or a saponin having a quillaic acid aglycone core, a Gal-(1→2)-[Xyl-(1→3)]-GlcA carbohydrate substituent at the C-3 beta-OH group, and a Glc-(1→3)-Xyl-(1→4)-Rha-(1→2)-[Xyl-(1→3)-4-OAc-Qui-(1→4)]-Fuc carbohydrate substituent at the C-28-OH group. and / or 3-O-beta-D-galactopyranosyl-(1→2)-[beta-D-xylopyranosyl-(1→3)]-beta-D-glucuronopyranosylquilayic acid, 28-O-beta-D-glucopyranosyl-(1→3)-beta-D-xylopyranosyl-(1→4)-alpha-L-rhamnopyranosyl-(1→2)-[beta-D-xylopyranosyl-(1→3)-4-OAc-beta-D-quinovopyranosyl-(1→4)]-beta-D-fucopyranoside. More preferably, the saponin is SO1861 and / or QS-21.

[0238] An embodiment of the third series of aspects and embodiments of the present invention is an effector moiety according to the present invention, wherein the saponin is a bisdesmosidic saponin and has a molecular mass of at least 1,500 daltons and comprises an oleanane-type triterpene containing an aldehyde group at the C-23 position and optionally a hydroxyl group at the C-16 position, and a first branched carbohydrate side chain at the C-3 position, which optionally contains glucuronic acid. The saponin comprises an ester group with a second branched carbohydrate side chain at the C-28 position, the second branched carbohydrate chain preferably comprising at least four carbohydrate units, optionally comprising at least one acetyl residue, e.g., two acetyl residues, and / or optionally comprising one or more deoxycarbohydrates and / or quinovose and / or glucose and / or 4-methoxycinnamic acid, and / or optionally comprising 5-O-[5-O-Ara / Api-3,5-dihydroxy-6-methyl-octanoyl]-3,5-dihydroxy-6-methyl-octanoic acid, and / or optionally comprising 5-O-[5-O-Rha-(1→2)-Ara / Api-3,5-dihydroxy-6-methyl-octanoyl]-3,5-dihydroxy-6-methyl-octanoic acid, linked to the carbohydrate via an ester bond. Alternatively, the at least one saponin is QS-21 or any one or more of QS-21A, QS-21 A-api, QS-21 A-xyl, QS-21B, QS-21 B-api, QS-21 B-xyl, QS-7-xyl, QS-7-api, QS-17-api, QS-17-xyl, QS-18, QS-1861, protonated QS1861 (QS1862), and Quil-A.

[0239] One embodiment of the third series of aspects and embodiments of the present invention is an effector moiety according to the present invention, wherein at least one saponin is a bisdesmosidic triterpene saponin belonging to the 12,13-dehydrooleanane type having an aldehyde functional group at position C-23. When present, the at least one saponin is covalently coupled to an amino acid residue of the effector moiety via an aldehyde functional group on the saponin, preferably at position C-23, preferably via at least one linker, more preferably via at least one cleavable linker. The amino acid residue is preferably selected from cysteine ​​and lysine.

[0240] One embodiment of the third series of aspects and embodiments of the present invention is an effector moiety according to the present invention, wherein at least one saponin is a bisdesmosidic triterpene saponin belonging to the 12,13-dehydrooleanane type having an aldehyde functional group at position C-23 and a glucuronic acid functional group on the carbohydrate substituent at the C-3 beta-OH group of the saponin. When present, the at least one saponin is covalently coupled to an amino acid residue of the effector moiety via the glucuronic acid functional group on the carbohydrate substituent at the C-3 beta-OH group of the saponin, preferably via at least one linker. The amino acid residue is preferably selected from cysteine ​​and lysine.

[0241] An embodiment of the third series of aspects and embodiments of the invention is an effector moiety according to the invention, wherein at least one linker comprises at least one non-cleavable linker and / or at least one cleavable linker, optionally wherein said cleavable linker undergoes cleavage under acidic, reductive, enzymatic or photoinduced conditions. Preferably, the cleavable linker comprises a cleavable bond selected from hydrazone and hydrazide bonds that undergo cleavage under acidic conditions, and / or bonds that are sensitive to proteolysis, e.g., proteolysis by cathepsin B, and / or bonds that undergo cleavage under reductive conditions, e.g., disulfide bonds.

[0242] An embodiment of the third series of aspects and embodiments of the invention is an effector moiety according to the invention, wherein at least one linker comprises at least one cleavable linker, which undergoes cleavage in vivo under acidic conditions present in the endosomes and / or lysosomes of mammalian cells, preferably human cells, preferably at a pH between 4.0 and 6.5, more preferably at a pH ≦5.5.

[0243]

[0013] One embodiment of the third series of aspects and embodiments of the invention is an effector moiety according to the invention, wherein at least one saponin is covalently attached to a lysine side chain forming an amide bond and / or to a cysteine ​​side chain forming a thio-ether linkage or a disulfide bond. The lysine and / or cysteine ​​is comprised by the effector moiety. The saponin is attached to the effector moiety directly or via at least one linker.

[0244] An embodiment of the third series of aspects and embodiments of the invention is an effector moiety according to the invention, wherein at least one saponin is covalently attached to the effector moiety via at least one linker, the linker being or comprising a scaffold comprising a polymeric or oligomeric structure and chemical groups for covalent coupling of the scaffold to the effector moiety.

[0245] An embodiment of the third series of aspects and embodiments of the invention is an effector moiety according to the invention, wherein at least one saponin is covalently attached to the backbone polymeric or oligomeric structure via a cleavable bond.

[0246] An embodiment of the third series of aspects and embodiments of the invention is an effector moiety according to the invention, wherein the cleavable bond is subject to cleavage under any of acidic, reductive, enzymatic and photoinduced conditions. More preferably, the cleavable bond is a hydrazone or hydrazide bond that is subject to cleavage under acidic conditions, and / or a bond that is susceptible to proteolysis, e.g., proteolysis by cathepsin B, and / or a bond that is subject to cleavage under reductive conditions, e.g., a disulfide bond.

[0247] An embodiment of the third series of aspects and embodiments of the invention is an effector moiety according to the invention, wherein the cleavable bond undergoes cleavage in vivo under acidic conditions present in the endosomes and / or lysosomes of mammalian cells, preferably human cells, preferably at a pH between 4.0 and 6.5, more preferably at a pH ≦ 5.5.

[0248] An embodiment of the third series of aspects and embodiments of the present invention is an effector moiety according to the present invention, wherein at least one saponin is covalently attached to a backbone polymeric or oligomeric structure via an imine, hydrazone, hydrazide, oxime, 1,3-dioxolane, disulfide, thio-ether, amide, peptide and / or ester bond, preferably via at least one linker, preferably an amide, hydrazide, thio-ether and / or hydrazone bond.

[0249] One embodiment of the third series of aspects and embodiments of the invention is an effector moiety according to the invention, wherein at least one saponin is covalently attached to a backbone polymeric or oligomeric structure, and, if present, at least one aldehyde functional group at position C-23 of the saponin is involved in the covalent bond, which covalent bond is preferably an imine bond, or a hydrazone bond, or an amide bond, or a thio-ether bond, or a disulfide bond, and / or, if present, at least one glucuronic acid functional group on a carbohydrate substituent at the C-3 beta-OH group of the saponin is involved in the covalent bond, which covalent bond is preferably an amide bond, or a disulfide bond, or a thio-ether bond.

[0250] One embodiment of the third series of aspects and embodiments of the invention is an effector moiety according to the invention, wherein the aldehyde functionality at position C-23 of at least one saponin is covalently coupled to a linker N-ε-maleimidocaproic acid hydrazide, which is covalently coupled to a sulfhydryl group, such as a cysteine ​​sulfhydryl group, on the backbone polymeric or oligomeric structure via a thio-ether bond.

[0251] An embodiment of this third series of aspects and embodiments of the invention is an effector moiety according to the invention, wherein the glucuronic acid functionality in the carbohydrate substituent at the C-3 beta-OH group of at least one saponin is covalently attached to the linker 1-[bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium 3-oxide hexafluorophosphate, which linker is covalently coupled via an amide bond to an amine group on a backbone polymer or oligomeric structure, for example to the amine group of a polymer or N-terminal lysine of the backbone polymer or oligomeric structure.

[0252] An embodiment of the third series of aspects and embodiments of the present invention is an effector moiety according to the present invention, wherein the chemical group of the scaffold for covalent coupling of the scaffold to the effector moiety is a click chemistry group, preferably selected from tetrazine, azide, alkene or alkyne, or cyclic derivatives of these groups, more preferably the click chemistry group is azide.

[0253] An embodiment of the third series of aspects and embodiments of the present invention is an effector moiety according to the present invention, wherein the backbone polymeric or oligomeric structure comprises a linear, branched and / or cyclic polymer, oligomer, dendrimer, dendron, dendronized polymer, dendronized oligomer, DNA, polypeptide, polylysine, polyethylene glycol, or an assembly of these polymeric or oligomeric structures, which assembly is preferably assembled by covalent crosslinking.

[0254] An embodiment of the third series of aspects and embodiments of the invention is an effector moiety according to the invention, wherein the at least one saponin is a defined number or range of saponins, preferably 1 to 128 saponins, or at least 2, 3, 4, 5, 6, 8, 10, 16, 32, 64, or 128 saponins, or any number therebetween, for example 7, 9, 12 saponins.

[0255] An embodiment of the third series of aspects and embodiments of the present invention is an effector moiety according to the present invention, wherein the effector moiety comprises more than one saponin, preferably 2, 3, 4, 5, 6, 8, 10, 16, 32, 64, or 1-100 saponins, or any number therebetween, for example 7, 9, 12 saponins, covalently attached directly to an amino acid residue of the effector moiety, preferably to a cysteine ​​and / or a lysine, and / or via at least one linker and / or via at least one cleavable linker, and / or via at least one polymeric or oligomeric backbone of any one of claims 14 to 23, preferably via 1 to 8 such backbones or 2 to 4 such backbones. Between 1 and 32 saponins, preferably 2, 3, 4, 5, 6, 8, 10, 16, or 32 saponins, are covalently attached to at least one backbone.

[0256] An aspect of the invention in the third series of aspects and embodiments of the invention relates to an antibody-drug conjugate comprising an effector moiety according to the invention or a ligand-drug conjugate comprising an effector moiety according to the invention.

[0257] In one embodiment of the third series of aspects and embodiments of the invention is an antibody-drug conjugate or a ligand-drug conjugate according to the invention, wherein the antibody is selected from the group consisting of CD71, CA125, EpCAM (17-1A), CD52, CEA, CD44v6, FAP, EGF-IR, integrin, syndecan-1, vascular integrin alpha-V beta-3, HER2, EGFR, CD20, CD22, folate receptor 1, CD146, CD56, CD19, CD138, CD27L receptor, PSMA, CanAg, integrin. The antibody may bind to any one of: leukocyte-alpha V, CA6, CD33, mesothelin, Cripto, CD3, CD30, CD239, CD70, CD123, CD352, DLL3, CD25, ephrin A4, MUC1, Trop2, CEACAM5, CEACAM6, HER3, CD74, PTK7, Notch3, FGF2, C4.4A, FLT3, CD38, FGFR3, CD7, PD-L1, CTLA4, CD52, PDGFRA, VEGFR1, VEGFR2, preferably CD71, HER2, and EGFR. and / or the antibody of the antibody-drug conjugate is or comprises any one of cetuximab, daratumumab, gemtuzumab, trastumumab, panitumumab, brentuximab, inotuzumab, moxetumomab, polatuzumab, obinutuzumab, OKT-9 anti-CD71 monoclonal antibody of the IgG type, pertuzumab, rituximab, ofatumumab, herceptin, alemtuzumab, pinatuzumab, OKT-10 anti-CD38 monoclonal antibody, an antibody of Table A2 or Table A3 or Table A4, preferably cetuximab or trastuzumab or OKT-9, or at least one tumor cell-specific receptor-binding fragment thereof and / or at least one tumor cell-specific receptor-binding domain thereof. and / or the antibody-drug conjugate comprises gemtuzumab ozogamicin, brentuximab vedotin, trastuzumab emtansine, inotuzumab ozogamicin, moxetumomab pasudotox, and polatuzumab vedotin, and any one of the antibody-drug conjugates of Table A2 and Table A3.And / or the ligand-drug conjugate comprises or consists of at least one non-proteinaceous ligand and / or at least one proteinaceous ligand for binding to a cell surface molecule such as EGF or a cytokine.

[0258] Certain aspects of the invention in the third series of aspects and embodiments of the invention relate to therapeutic combinations comprising: (a) an effector moiety according to the invention, and optionally a pharmaceutically acceptable excipient; and (b) an antibody-drug conjugate or a ligand-drug conjugate, and optionally a pharmaceutically acceptable excipient.

[0259] In one embodiment of the third series of aspects and embodiments of the invention is a therapeutic combination according to the invention, wherein the antibody-drug conjugate is selected from the group consisting of tumor cell receptors CD71, CA125, EpCAM (17-1A), CD52, CEA, CD44v6, FAP, EGF-IR, integrins, syndecan-1, vascular integrin alpha-V beta-3, HER2, EGFR, CD20, CD22, folate receptor 1, CD146, CD56, CD19, CD138, CD27L receptor, PSMA, CanAg, integrins - capable of binding to any one of alpha V, CA6, CD33, mesothelin, Cripto, CD3, CD30, CD239, CD70, CD123, CD352, DLL3, CD25, ephrin A4, MUC1, Trop2, CEACAM5, CEACAM6, HER3, CD74, PTK7, Notch3, FGF2, C4.4A, FLT3, CD38, FGFR3, CD7, PD-L1, CTLA4, CD52, PDGFRA, VEGFR1, VEGFR2, preferably CD71, HER2, EGFR. and / or the antibody of the antibody-drug conjugate is or comprises any one of cetuximab, daratumumab, gemtuzumab, trastumumab, panitumumab, brentuximab, inotuzumab, moxetumomab, polatuzumab, obinutuzumab, OKT-9 anti-CD71 monoclonal antibody of the IgG type, pertuzumab, rituximab, ofatumumab, herceptin, alemtuzumab, pinatuzumab, OKT-10 anti-CD38 monoclonal antibody, an antibody of Table A2 or Table A3 or Table A4, preferably cetuximab or trastuzumab or OKT-9, or at least one tumor cell-specific receptor-binding fragment thereof and / or at least one tumor cell-specific receptor-binding domain thereof. and / or the antibody-drug conjugate comprises gemtuzumab ozogamicin, brentuximab vedotin, trastuzumab emtansine, inotuzumab ozogamicin, moxetumomab pasudotox, and polatuzumab vedotin, and any one of the antibody-drug conjugates of Table A2 and Table A3.And / or the ligand-drug conjugate comprises or consists of at least one non-proteinaceous ligand and / or at least one proteinaceous ligand for binding to a cell surface molecule such as EGF or a cytokine.

[0260] An aspect of the invention in the third series of aspects and embodiments of the invention relates to a pharmaceutical composition comprising an effector moiety according to the invention, or an antibody-drug conjugate according to the invention, or a ligand-drug conjugate according to the invention, and optionally a pharmaceutically acceptable excipient.

[0261] An aspect of the invention in the third series of aspects and embodiments of the invention relates to an effector moiety according to the invention, or an antibody-drug conjugate according to the invention, or a therapeutic combination according to the invention, or a ligand-drug conjugate according to the invention, or a pharmaceutical composition according to the invention, for use as a medicament.

[0262] An aspect of the invention in the third series of aspects and embodiments of the invention relates to an effector moiety according to the invention, or an antibody-drug conjugate according to the invention, or a ligand-drug conjugate according to the invention, or a therapeutic combination according to the invention, or a pharmaceutical composition according to the invention, for use in the treatment or prevention of cancer or an autoimmune disease.

[0263] An aspect of the invention in the fourth series of aspects and embodiments of the invention relates to a conjugate comprising a cell surface targeting molecule and at least one effector moiety, and further comprising at least one covalently attached saponin.

[0264] An embodiment of the fourth series of aspects and embodiments of the invention is a conjugate of the invention, wherein at least one saponin is a triterpenoid saponin or a bisdesmosidic triterpene saponin belonging to the 12,13-dehydrooleanane type having an aldehyde functional group at position C-23 and optionally a glucuronic acid functional group in the carbohydrate substituent of the C-3 beta-OH group of the saponin, and / or a saponin isolated from Gypsophila species and / or Saponaria species and / or Agrostemma species and / or Quillaja species, e.g. Quillaja saponaria.

[0265] An embodiment of the fourth set of aspects and embodiments of the invention is a conjugate of the invention, wherein the at least one saponin is a single specific saponin or a mixture of two or more different saponins.

[0266] An embodiment of the fourth set of aspects and embodiments of the invention is a conjugate of the invention, wherein at least one saponin has a molecular mass of 3,000 daltons or less, preferably 2,500 daltons or less, more preferably 2,300 daltons or less, most preferably 2,000 daltons or less, for example between 1,500 and 1,900 daltons.

[0267]

[0023] In one embodiment of the fourth series of aspects and embodiments of the invention is a conjugate of the invention, wherein the at least one saponin is one or more of the saponins of Table A1 or Scheme I, SO1861, SA1657, GE1741, SA1641, QS-21, QS-21A, QS-21 A-api, QS-21 A-xyl, QS-21B, QS-21 B-api, QS-21 B-xyl, QS-7-xyl, QS-7-api, QS-17-api, QS-17-xyl, QS1861, QS1862, Quillaja saponin, Saponinum albumenum album, QS-18, Quil-A, Gyp1, gypsoside A, AG1, AG2, SO1542, SO1584, SO1658, SO1674, SO1832, or any stereoisomer thereof and / or any combination thereof. Preferably, the saponin is SO1861 and / or GE1741 and / or SA1641 and / or QS-21, and / or a saponin having a quillaic acid aglycone core, a Gal-(1→2)-[Xyl-(1→3)]-GlcA carbohydrate substituent at the C-3 beta-OH group, and a Glc-(1→3)-Xyl-(1→4)-Rha-(1→2)-[Xyl-(1→3)-4-OAc-Qui-(1→4)]-Fuc carbohydrate substituent at the C-28-OH group. and / or 3-O-beta-D-galactopyranosyl-(1→2)-[beta-D-xylopyranosyl-(1→3)]-beta-D-glucuronopyranosylquilayic acid, 28-O-beta-D-glucopyranosyl-(1→3)-beta-D-xylopyranosyl-(1→4)-alpha-L-rhamnopyranosyl-(1→2)-[beta-D-xylopyranosyl-(1→3)-4-OAc-beta-D-quinovopyranosyl-(1→4)]-beta-D-fucopyranoside. More preferably, the saponin is SO1861 and / or QS-21.

[0268] An embodiment of the fourth set of aspects and embodiments of the invention is a conjugate of the invention, wherein at least one saponin is a bisdesmosidic saponin and comprises an oleanane-type triterpene having a molecular mass of at least 1,500 daltons and containing an aldehyde group at the C-23 position and optionally a hydroxyl group at the C-16 position, and a first branched carbohydrate side chain at the C-3 position, which first branched carbohydrate side chain optionally contains glucuronic acid. The saponin contains an ester group with a second branched carbohydrate side chain at the C-28 position, this second branched carbohydrate chain preferably containing at least four carbohydrate units, optionally containing at least one acetyl residue, for example two acetyl residues, and / or optionally containing a deoxy carbohydrate, and / or optionally containing quinovose, and / or optionally containing glucose, and / or optionally containing 4-methoxycinnamic acid, and / or optionally containing 5-O-[5-O-Ara / Api-3,5-dihydroxy-6-methyl-octanoyl]-3,5-dihydroxy-6-methyl-octanoic acid, and / or optionally containing 5-O-[5-O-Rha-(1→2)-Ara / Api-3,5-dihydroxy-6-methyl-octanoyl]-3,5-dihydroxy-6-methyl-octanoic acid, linked to the carbohydrate via an ester bond. Alternatively, the at least one saponin is QS-21 or any one or more of QS-21A, QS-21 A-api, QS-21 A-xyl, QS-21B, QS-21 B-api, QS-21 B-xyl, QS-7-xyl, QS-7-api, QS-17-api, QS-17-xyl, QS-18, QS-1861, protonated QS1861 (QS1862), and Quil-A.

[0269] An embodiment of the fourth series of aspects and embodiments of the invention is a conjugate of the invention, wherein the cell surface molecule targeting molecule comprises or consists of a ligand, proteinaceous ligand, or proteinaceous binding molecule for binding to the cell surface molecule.

[0270] An embodiment of the fourth series of aspects and embodiments of the invention is a conjugate of the invention, wherein the cell surface molecule targeting molecule comprises or consists of a non-proteinaceous and / or proteinaceous ligand for binding to a cell surface molecule such as EGF or a cytokine, and / or comprises or consists of an immunoglobulin, at least one binding domain of an immunoglobulin, and / or at least one binding fragment of an immunoglobulin, such as an antibody, an IgG, a molecule comprising or consisting of a Vhh domain or a Vh domain, a Fab, scFv, Fv, dAb, F(ab)2, Fcab fragment, which is capable of binding to a cell surface molecule.

[0271] An embodiment of the fourth series of aspects and embodiments of the invention is a conjugate of the invention, wherein the cell surface molecule targeting molecule is a tumor cell surface molecule, preferably a tumor cell receptor, such as a tumor cell specific receptor, more preferably CD71, CA125, EpCAM (17-1A), CD52, CEA, CD44v6, FAP, EGF-IR, integrin, syndecan-1, vascular integrin alpha-V beta-3, HER2, EGFR, CD20, CD22, folate receptor 1, CD146, CD56, CD19, CD138, CD27L receptor, PSMA. , CanAg, integrin-alpha V, CA6, CD33, mesothelin, Cripto, CD3, CD30, CD239, CD70, CD123, CD352, DLL3, CD25, ephrin A4, MUC1, Trop2, CEACAM5, CEACAM6, HER3, CD74, PTK7, Notch3, FGF2, C4.4A, FLT3, CD38, FGFR3, CD7, PD-L1, CTLA4, CD52, PDGFRA, VEGFR1, VEGFR2, preferably CD71, HER2, and EGFR.

[0272] An embodiment of the fourth series of aspects and embodiments of the invention is a conjugate of the invention, wherein the tumor cell receptor is internalized by the tumor cell after binding to the cell surface molecule targeting molecule of the invention and the conjugate of the invention. Preferably, binding of the conjugate to the tumor cell receptor is followed by internalization of the complex of the conjugate and the tumor cell receptor mediated by the tumor cell receptor, for example via endocytosis.

[0273] An embodiment of the fourth set of aspects and embodiments of the invention is a conjugate of the invention, wherein the cell surface molecule targeting molecule is or comprises a monoclonal antibody or at least one cell surface molecule binding fragment or domain thereof, preferably comprising or consisting of cetuximab, daratumumab, gemtuzumab, trastuzumab, panitumumab, brentuximab, inotuzumab, moxetumomab, polatuzumab, obinutuzumab, OKT-9 anti-CD71 monoclonal antibody of the IgG type, pertuzumab, rituximab, ofatumumab, herceptin, alemtuzumab, pinatuzumab, OKT-10 anti-CD38 monoclonal antibody, any one of the antibodies of Table A2 or Table A3 or Table A4, preferably cetuximab or trastuzumab or OKT-9, or at least one cell surface molecule binding fragment or domain thereof.

[0274] An embodiment of the fourth series of aspects and embodiments of the invention is a conjugate of the invention, wherein at least one effector moiety comprises or consists of any one or more of an oligonucleotide, a nucleic acid, and a xenonucleic acid, preferably a vector, a gene, a transgene that induces cell suicide, deoxyribonucleic acid (DNA), ribonucleic acid (RNA), antisense oligonucleotide (ASO, AON), short interfering RNA (siRNA), microRNA (miRNA), a DNA aptamer, an RNA aptamer, mRNA, minicircle DNA, peptide nucleic acid ( PNA), phosphoramidate morpholino oligomer (PMO), locked nucleic acid (LNA), bridged nucleic acid (BNA), 2'-deoxy-2'-fluoroarabinonucleic acid (FANA), 2'-O-methoxyethyl-RNA (MOE), 2'-O,4'-aminoethylene-bridged nucleic acid, 3'-fluorohexitol nucleic acid (FHNA), plasmid, glycol nucleic acid (GNA), and threose nucleic acid (TNA), or derivatives thereof, more preferably BNA, for example, BNA for silencing HSP27 protein expression.

[0275] An embodiment of the fourth series of aspects and embodiments of the invention is a conjugate of the invention, wherein the at least one effector moiety comprises or consists of at least one proteinaceous molecule, preferably selected from peptides, proteins, enzymes such as urease and Cre recombinase, ribosome-inactivating proteins, proteinaceous toxins selected from Table A5, more preferably viral toxins such as apoptin; bacterial toxins such as Shiga toxins, Shiga-like toxins, Pseudomonas The protein toxin is selected from any one or more of: P. aeruginosa exotoxin (PE), or exotoxin A of PE, full-length or truncated diphtheria toxin (DT), cholera toxin; fungal toxins, such as alpha-sarcin; plant toxins, including ribosome-inactivating proteins and type 2 ribosome-inactivating protein A chains, such as giantin-30 or giantin-32, saporin-S3 or saporin-S6, bouganin or the deimmunized derivative of bouganin, debouganin, Shiga-like toxin A, pokeweed antiviral protein, ricin, ricin A chain, modeccin, modeccin A chain, abrin, abrin A chain, volkensin, volkensin A chain, viscumin, viscumin A chain; or animal or human toxins, such as frog RNase, or granzyme B or angiogenin from humans, or any fragment or derivative thereof. Preferably, the protein toxin is giantin and / or saporin.

[0276] An embodiment of the fourth series of aspects and embodiments of the invention is a conjugate of the invention, wherein the at least one effector moiety comprises or consists of at least one payload, preferably any one or more of a ribosome-targeting toxin, an elongation factor-targeting toxin, a tubulin-targeting toxin, a DNA-targeting toxin, and an RNA-targeting toxin, more preferably emtansine, pasudotox, maytansinoid derivative DM1, maytansinoid derivative DM4, monomethyl auristatin E (MMAE, vedotin), monomethyl auristatin F (MMAF, mafodotin), Calicheamicin, N-acetyl-γ-calicheamicin, pyrrolobenzodiazepine (PBD) dimer, benzodiazepine, CC-1065 analog, duocarmycin, doxorubicin, paclitaxel, docetaxel, cisplatin, cyclophosphamide, etoposide, docetaxel, 5-fluorouracil (5-FU), mitoxantrone, tubulysin, indolinobenzodiazepine, AZ13599185, cryptophycin, rhizoxin, methotrexate, anthracycline, camptothecin analog, SN-38, DX-8951f, exatecan mesylate, Pseudomonas The steroid inhibitor is selected from one or more of a truncated form of P. aeruginosa exotoxin (PE38), a duocarmycin derivative, amanitin, α-amanitin, splicostatin, tylanstatin, ozogamicin, tesirin, amberstatin 269, and soravtansine, or a derivative thereof.

[0277] An embodiment of the fourth series of aspects and embodiments of the invention is a conjugate of the invention, wherein at least one effector moiety is covalently attached to the cell surface molecule targeting molecule via at least one linker or directly attached to the cell surface molecule targeting molecule.

[0278] An embodiment of the fourth set of aspects and embodiments of the invention is a conjugate of the invention, wherein at least one effector moiety is covalently attached to a cell surface targeting molecule, thereby forming the antibody-drug conjugate gemtuzumab ozogamicin, brentuximab vedotin, trastuzumab emtansine, inotuzumab ozogamicin, moxetumomab pasudotox, and polatuzumab vedotin, and any one of the antibody-drug conjugates of Table A2 and Table A3.

[0279] An embodiment of the fourth series of aspects and embodiments of the invention is a conjugate of the invention, wherein at least one saponin is covalently attached to a cell surface molecular targeting molecule, preferably an amino acid residue of the cell surface molecular targeting molecule, via an aldehyde functional group on the saponin, and / or to at least one effector moiety, preferably via an amino acid residue on the at least one effector moiety, via an aldehyde functional group on the saponin, preferably an aldehyde functional group at position C-23 of a bisdesmosidic triterpene saponin belonging to the 12,13-dehydrooleanane type.

[0280] In one embodiment of the fourth series of aspects and embodiments of the invention, the conjugate of the invention comprises an aldehyde functional group of at least one saponin, preferably an aldehyde functional group at position C-23 of at least one saponin, covalently attached to a linker, N-ε-maleimidocaproic acid hydrazide, which is covalently coupled via a thio-ether bond to a sulfhydryl group, e.g., a sulfhydryl group of a cysteine, on a cell surface targeting molecule and / or at least one effector moiety.

[0281] One embodiment of the fourth series of aspects and embodiments of the invention is a conjugate of the invention, wherein at least one saponin is a bisdesmosidic triterpene saponin belonging to the 12,13-dehydrooleanane type, having an aldehyde functional group at position C-23 and comprising a glucuronic acid functional group on a carbohydrate substituent at the C-3 beta-OH group of the saponin, and the saponin is covalently attached to an amino acid residue of a cell surface targeting molecule and / or to at least one effector moiety via said glucuronic acid functional group, preferably via an amino acid residue of at least one effector moiety.

[0282] In one embodiment of this fourth series of aspects and embodiments of the invention, a conjugate of the invention is provided in which a glucuronic acid functional group on the carbohydrate substituent at the C-3 beta-OH group of at least one saponin is covalently coupled to the linker 1-[bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium 3-oxide hexafluorophosphate, which is covalently coupled via an amide bond to an amine group on the cell surface molecule targeting molecule and / or at least one effector moiety, for example, a lysine or N-terminal amine group of the cell surface molecule targeting molecule and / or at least one effector moiety.

[0283] An embodiment of the fourth series of aspects and embodiments of the invention is a conjugate of the invention, wherein at least one saponin is covalently attached to a cell surface molecular targeting molecule and / or at least one effector moiety either directly or via at least one linker, such as a bifunctional linker based on N-ε-maleimidocaproic acid hydrazide and / or 1-[bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium 3-oxide hexafluorophosphate, or a trifunctional linker, for example the trifunctional linker of Scheme II.

[0284] An embodiment of the fourth series of aspects and embodiments of the invention is a conjugate of the invention, wherein the trifunctional linker comprises a second chemical group having at least one saponin covalently attached thereto, a third chemical group for covalently attaching to a cell surface targeting molecule, and a first chemical group for covalently attaching to at least one effector moiety. Preferably, the trifunctional linker is the trifunctional linker of Scheme II and Structure B.

[0285] One embodiment of this fourth series of aspects and embodiments of the invention is a conjugate of the invention, wherein at least one saponin is covalently attached to a cell surface molecule targeting molecule and to at least one effector moiety via at least one linker comprising a trifunctional linker, to which both the cell surface molecule targeting molecule and the at least one effector moiety are attached. Preferably, the trifunctional linker is the trifunctional linker of Scheme II and Structure B.

[0286]

[0013] An embodiment of the fourth series of aspects and embodiments of the invention is a conjugate of the invention, wherein at least one linker comprises at least one cleavable linker. Optionally, said cleavable linker undergoes cleavage under acidic, reductive, enzymatic, or photoinduced conditions. Preferably, the cleavable linker comprises a cleavable bond selected from hydrazone and hydrazide bonds that undergo cleavage under acidic conditions, and / or bonds that are sensitive to proteolysis, e.g., proteolysis by cathepsin B, and / or bonds that undergo cleavage under reductive conditions, e.g., disulfide bonds.

[0287] An embodiment of the fourth set of aspects and embodiments of the invention is a conjugate of the invention, wherein at least one linker comprises at least one cleavable linker, said cleavable linker undergoing cleavage in vivo under acidic conditions present in endosomes and / or lysosomes of mammalian cells, preferably human cells, preferably at a pH between 4.0 and 6.5, more preferably at a pH ≦ 5.5.

[0288]

[0013] In one embodiment of this fourth series of aspects and embodiments of the present invention, the conjugate of the present invention comprises at least one saponin covalently attached to a lysine side chain forming an amide bond and / or to a cysteine ​​side chain forming a thio-ether linkage or a disulfide bond. The lysine and / or cysteine ​​are comprised by a cell surface targeting molecule and / or by at least one effector moiety. The at least one saponin is attached directly to the lysine and / or cysteine ​​or via at least one linker and / or trifunctional linker, optionally including a cleavable linker, such as the trifunctional linker of Scheme II and Structure B.

[0289] An embodiment of the fourth series of aspects and embodiments of the invention is a conjugate of the invention, wherein the linker is based on N-ε-maleimidocaproic acid hydrazide, and / or 1-[bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium 3-oxide hexafluorophosphate, a trifunctional linker, such as the trifunctional linker of Scheme II and Structure B, a cleavable linker, and / or involves one or more of a disulfide bond, a thio-ether bond, an amide bond, a hydrazide bond.

[0290] An embodiment of the fourth set of aspects and embodiments of the invention is a conjugate of the invention, wherein at least one saponin is covalently attached to a cell surface molecule targeting molecule and / or at least one effector moiety via at least one linker, the linker being or comprising a scaffold comprising a polymeric or oligomeric structure and further comprising at least one fourth chemical group for covalently coupling the scaffold to the cell surface molecule targeting molecule and / or at least one effector moiety.

[0291] An embodiment of the fourth series of aspects and embodiments of the invention is a conjugate of the invention, wherein at least one saponin is covalently attached to a backbone polymeric or oligomeric structure via a cleavable bond and / or via a non-cleavable bond.

[0292] An embodiment of the fourth series of aspects and embodiments of the invention is a conjugate of the invention, wherein the cleavable bond undergoes cleavage under any of acidic, reductive, enzymatic, and photoinduced conditions. Preferably, the cleavable bond comprises a hydrazone bond and a hydrazide bond that undergoes cleavage under acidic conditions, and / or a bond that is susceptible to proteolysis, e.g., proteolysis by cathepsin B, and / or a bond that undergoes cleavage under reductive conditions, e.g., a disulfide bond.

[0293] An embodiment of the fourth set of aspects and embodiments of the invention is a conjugate of the invention, wherein the cleavable bond undergoes cleavage in vivo under acidic conditions present in the endosomes and / or lysosomes of mammalian cells, preferably human cells, preferably at a pH between 4.0 and 6.5, more preferably at a pH ≦ 5.5.

[0294] An embodiment of the fourth series of aspects and embodiments of the invention is a conjugate of the invention, wherein at least one saponin is covalently attached to a backbone polymeric or oligomeric structure via any one or more of an imine bond, hydrazone bond, hydrazide bond, oxime bond, 1,3-dioxolane bond, disulfide bond, thio-ether bond, amide bond, peptide bond or ester bond, preferably via at least one linker.

[0295] One embodiment of the fourth series of aspects and embodiments of the invention is a conjugate of the invention, wherein at least one saponin is covalently attached to a backbone polymeric or oligomeric structure via any one or more of an imine bond, a hydrazone bond and a hydrazide bond, which bond is preferably cleavable in accordance with the invention, and preferably the at least one saponin is covalently attached to the backbone polymeric or oligomeric structure via an aldehyde functionality at position C-23 of the at least one saponin.

[0296] One embodiment of the fourth series of aspects and embodiments of the invention is a conjugate of the invention, wherein the aldehyde functionality at position C-23 of at least one saponin is covalently attached to a linker N-ε-maleimidocaproic acid hydrazide, which is covalently coupled to a sulfhydryl group, such as a cysteine ​​sulfhydryl group, on a backbone polymeric or oligomeric structure via a thio-ether bond.

[0297] An embodiment of the fourth series of aspects and embodiments of the invention is a conjugate of the invention, wherein at least one saponin is covalently attached to the backbone polymeric or oligomeric structure via an amide bond. Preferably, at least one saponin, when present, is covalently attached to the backbone polymeric or oligomeric structure via a glucuronic acid functionality on a carbohydrate substituent of the C-3 beta-OH group of at least one saponin.

[0298] An embodiment of the fourth series of aspects and embodiments of the invention is a conjugate of the invention, wherein a glucuronic acid functionality in the carbohydrate substituent at the C-3 beta-OH group of at least one saponin is covalently attached to the linker 1-[bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium 3-oxide hexafluorophosphate, which is covalently coupled via an amide bond to an amine group on a backbone polymer or oligomeric structure, for example to the amine group of a polymer or an N-terminal lysine of the backbone polymer or oligomeric structure.

[0299] One embodiment of the fourth series of aspects and embodiments of the invention is a conjugate of the invention, wherein at least one saponin is covalently attached to the backbone polymeric or oligomeric structure, if present, via an aldehyde functionality at position C-23 of at least one saponin, and / or, if present, via a glucuronic acid functionality on a carbohydrate substituent of the C-3 beta-OH group of at least one saponin.

[0300] An embodiment of the fourth series of aspects and embodiments of the invention is a conjugate of the invention, wherein at least one fourth chemical group of the scaffold for covalent coupling of the scaffold to a cell surface targeting molecule and / or to at least one effector moiety is a click chemistry group, preferably selected from any one or more of a tetrazine, an azide, an alkene, or an alkyne, or cyclic derivatives of these groups, preferably an azide group.

[0301] An embodiment of the fourth series of aspects and embodiments of the invention is a conjugate of the invention, wherein the backbone polymeric or oligomeric structure comprises a linear, branched, and / or cyclic polymer, oligomer, dendrimer, dendron, dendronized polymer, dendronized oligomer, DNA, polypeptide, polylysine, polyethylene glycol, or an assembly of these polymeric or oligomeric structures, which assembly is preferably assembled by covalent crosslinking.

[0302] An embodiment of the fourth set of aspects and embodiments of the invention is a conjugate of the invention, wherein the at least one saponin is a defined number or range of saponins, preferably 1 to 128 saponins, or at least 2, 3, 4, 5, 6, 8, 10, 16, 32, 64, or 128 saponins, or any number therebetween, for example 7, 9, 12 saponins.

[0303] An embodiment of the fourth set of aspects and embodiments of the invention is a conjugate of the invention, wherein the conjugate comprises more than one saponin, preferably 2, 3, 4, 5, 6, 8, 10, 16, 32, 64, or 1-100 saponins, or any number of saponins in between, for example 7, 9, 12 saponins, covalently attached directly to an amino acid residue of a cell surface targeting molecule and / or to at least one effector moiety, preferably via an amino acid residue on at least one effector moiety, preferably to a cysteine ​​and / or lysine, and / or via at least one linker and / or via at least one cleavable linker, and / or to at least one polymeric or oligomeric backbone of the invention, preferably via 1 to 8 such backbones or 2 to 4 such backbones. Between 1 and 32 saponins, for example, 2, 3, 4, 5, 6, 8, 10, 16, or 32 saponins, or any number therebetween, for example, 7, 9, 12 saponins, are covalently attached to at least one backbone.

[0304] One embodiment of the fourth series of aspects and embodiments of the present invention is a conjugate of the present invention, in which at least one saponin is covalently linked to a cell surface molecule targeting molecule and to at least one effector moiety via a trifunctional linker. The trifunctional linker comprises a second chemical group to which at least one saponin is covalently linked, either directly or via a linker such as a cleavable linker and / or via a backbone comprising a polymeric or oligomeric structure, and a fourth chemical group according to the present invention for covalently coupling the backbone to the trifunctional linker. The trifunctional linker further comprises a third chemical group for covalently linking to the cell surface molecule targeting molecule and a first chemical group for covalently linking to at least one effector moiety. The cell surface molecule targeting molecule is linked to the third chemical group, and at least one effector moiety is linked to the first chemical group. Preferably, the trifunctional linker is the trifunctional linker of Scheme II and Structure B.

[0305] An aspect of the invention of the fourth series of aspects and embodiments of the invention relates to a pharmaceutical composition, comprising a conjugate of the invention and optionally a pharmaceutically acceptable excipient and / or a pharmaceutically acceptable diluent.

[0306] An aspect of the invention of the fourth series of aspects and embodiments of the invention relates to a conjugate of the invention or a pharmaceutical composition of the invention for use as a medicament.

[0307] An aspect of the invention in the fourth series of aspects and embodiments of the invention relates to a conjugate of the invention or a pharmaceutical composition of the invention for use in the treatment or prevention of cancer or an autoimmune disease.

[0308] One aspect of the invention within a fifth set of aspects and embodiments of the invention relates to a therapeutic combination for use as a pharmaceutical, comprising: (a) a fourth pharmaceutical composition comprising a fourth proteinaceous molecule comprising a binding site for binding to an epitope on a cell surface molecule, and at least one saponin covalently attached to said fourth proteinaceous molecule, preferably to an amino acid residue of said fourth proteinaceous molecule (the fourth pharmaceutical composition optionally further comprising a pharmaceutically acceptable excipient); and (b) a fifth pharmaceutical composition comprising a fifth proteinaceous molecule. The fifth proteinaceous molecule comprises a binding site for binding to the epitope on the cell surface molecule of (a) and an effector moiety. The fifth pharmaceutical composition optionally further comprises a pharmaceutically acceptable excipient. The binding site of the fourth proteinaceous molecule and the binding site of the fifth proteinaceous molecule are the same. The cell surface molecule and epitope on the cell surface molecule to which the fourth proteinaceous molecule can bind are the same as the cell surface molecule and epitope on the cell surface molecule to which the fifth proteinaceous molecule can bind.

[0309] An aspect of the invention of the fifth series of aspects and embodiments of the invention relates to the pharmaceutical composition of the fourth series of the invention for use as a medicament.

[0310] An aspect of the invention of the fifth series of aspects and embodiments of the invention relates to a therapeutic combination for use in treating or preventing cancer in a human subject, the therapeutic combination comprising (a) the fourth pharmaceutical composition of the invention and (b) the fifth pharmaceutical composition of the invention, wherein the cell surface molecule is expressed on the surface of a tumor cell, preferably the cell surface molecule is a tumor cell-specific surface molecule, and preferably the epitope is a tumor cell-specific epitope.

[0311] An aspect of the invention of the fifth series of aspects and embodiments of the invention relates to the pharmaceutical composition of the fourth series of the invention for use in treating or preventing cancer in a patient in need thereof, wherein the cell surface molecule is expressed on the surface of a tumor cell. Preferably, the cell surface molecule is a tumor cell-specific surface molecule, and preferably the epitope is a tumor cell-specific epitope.

[0312] An embodiment of the fifth set of aspects and embodiments of the invention is a fourth pharmaceutical composition for use according to the invention, or a combination of therapeutic agents for use according to the invention, wherein the fifth pharmaceutical composition of the invention and the fourth pharmaceutical composition are administered to a patient in need thereof.

[0313] An embodiment of the fifth set of aspects and embodiments of the invention is a therapeutic combination for use according to the invention, or a fourth pharmaceutical composition for use according to the invention, wherein the fourth proteinaceous molecule and the binding site of the fifth proteinaceous molecule comprise or consist of an immunoglobulin, or a binding fragment or binding domain of said immunoglobulin, such as an antibody, an IgG, a molecule comprising or consisting of a Vhh domain or a Vh domain, a Fab, scFv, Fv, dAb, F(ab)2, Fcab fragment, and / or comprise or consist of at least one ligand, a ligand for binding to a cell surface molecule such as EGF or a cytokine.

[0314] An embodiment of the fifth series of aspects and embodiments of the invention is a therapeutic combination for use according to the invention, or a fourth pharmaceutical composition for use according to the invention, wherein the tumor cell surface molecule is a cell surface receptor that is specifically present on tumor cells, and preferably the epitope is a tumor cell-specific epitope.

[0315] An embodiment of the fifth series of aspects and embodiments of the invention is a therapeutic combination for use according to the invention, or a fourth pharmaceutical composition for use according to the invention, wherein at least one saponin is a triterpenoid saponin or a bisdesmosidic triterpene saponin belonging to the 12,13-dehydrooleanane type having an aldehyde functional group at position C-23 and optionally a glucuronic acid functional group in the carbohydrate substituent of the C-3 beta-OH group of the saponin, and / or a saponin isolated from Gypsophila species and / or Saponaria species and / or Agrostemma species and / or Quillaja species, e.g. Quillaja saponaria.

[0316] Certain embodiments of the fifth set of aspects and embodiments of the invention are therapeutic combinations for use according to the invention, or fourth pharmaceutical compositions for use according to the invention, wherein the at least one saponin is a single particular saponin or a mixture of two or more different saponins, for example one or more of the saponins of Table A1 or Scheme I, SO1861, SA1657, GE1741, SA1641, QS-21, QS-21A, QS-21 A-api, QS-21 A-xyl, QS-21B, QS-21 B-api, QS-21 B-xyl, QS-7-xyl, QS-7-api, QS-17-api, QS-17-xyl, QS1861, QS1862, Quillaja saponin, Saponinum albumenum album, QS-18, Quil-A, Gyp1, gypsoside A, AG1, AG2, SO1542, SO1584, SO1658, SO1674, SO1832, or any stereoisomer thereof and / or any combination thereof. Preferably, the saponin is SO1861 and / or GE1741 and / or SA1641 and / or QS-21, and / or a saponin having a quillaic acid aglycone core, a Gal-(1→2)-[Xyl-(1→3)]-GlcA carbohydrate substituent at the C-3 beta-OH group, and a Glc-(1→3)-Xyl-(1→4)-Rha-(1→2)-[Xyl-(1→3)-4-OAc-Qui-(1→4)]-Fuc carbohydrate substituent at the C-28-OH group. and / or 3-O-beta-D-galactopyranosyl-(1→2)-[beta-D-xylopyranosyl-(1→3)]-beta-D-glucuronopyranosylquilayic acid, 28-O-beta-D-glucopyranosyl-(1→3)-beta-D-xylopyranosyl-(1→4)-alpha-L-rhamnopyranosyl-(1→2)-[beta-D-xylopyranosyl-(1→3)-4-OAc-beta-D-quinovopyranosyl-(1→4)]-beta-D-fucopyranoside. More preferably, the saponin is SO1861 and / or QS-21.

[0317] An embodiment of the fifth set of aspects and embodiments of the invention is a therapeutic combination for use according to the invention, or a fourth pharmaceutical composition for use according to the invention, wherein the at least one biologically active molecule is a bisdesmosidic saponin, comprising an oleanane-type triterpene having a molecular mass of at least 1,500 daltons and containing an aldehyde group at the C-23 position and optionally a hydroxyl group at the C-16 position, and having a first branched carbohydrate side chain at the C-3 position, which first branched carbohydrate side chain optionally contains glucuronic acid. The saponin contains an ester group at the C-28 position with a second branched carbohydrate side chain, which preferably contains at least four carbohydrate units, optionally containing at least one acetyl residue, for example two acetyl residues, and / or optionally at least one deoxy carbohydrate and / or quinovose and / or glucose and / or 4-methoxycinnamic acid, and / or optionally 5-O-[5-O-Ara / Api-3,5-dihydroxy-6-methyl-octanoyl]-3,5-dihydroxy-6-methyl-octanoic acid, and / or optionally 5-O-[5-O-Rha-(1→2)-Ara / Api-3,5-dihydroxy-6-methyl-octanoyl]-3,5-dihydroxy-6-methyl-octanoic acid, linked to the carbohydrate via an ester bond. Alternatively, the at least one saponin is QS-21 or any one or more of QS-21A, QS-21 A-api, QS-21 A-xyl, QS-21B, QS-21 B-api, QS-21 B-xyl, QS-7-xyl, QS-7-api, QS-17-api, QS-17-xyl, QS-18, QS-1861, protonated QS1861 (QS1862), and Quil-A.

[0318]

[0013] An embodiment of the fifth series of aspects and embodiments of the present invention is a therapeutic combination for use according to the present invention, or a fourth pharmaceutical composition for use according to the present invention, wherein at least one saponin is a bisdesmosidic triterpene saponin belonging to the 12,13-dehydroanane type having an aldehyde functional group at position C-23. The saponin is covalently coupled to a fourth proteinaceous molecule, preferably to an amino acid residue of the fourth proteinaceous molecule, via an aldehyde functional group on the saponin, preferably said aldehyde functional group at position C-23, preferably via at least one linker and / or at least one cleavable linker. The amino acid residue is preferably selected from cysteine ​​and lysine.

[0319] One embodiment of the fifth series of aspects and embodiments of the invention is a therapeutic combination for use according to the invention, or a fourth pharmaceutical composition for use according to the invention, wherein the aldehyde functionality at position C-23 of at least one saponin is covalently coupled to a linker N-ε-maleimidocaproic acid hydrazide, which is covalently coupled to a sulfhydryl group, such as a cysteine ​​sulfhydryl group, on a fourth proteinaceous molecule via a thio-ether bond.

[0320] One embodiment of the fifth series of aspects and embodiments of the present invention is a therapeutic combination for use according to the present invention, or a fourth pharmaceutical composition for use according to the present invention, wherein at least one saponin is a bisdesmosidic triterpene saponin belonging to the 12,13-dehydrooleanane type having an aldehyde functional group at position C-23 and containing a glucuronic acid functional group on the carbohydrate substituent at the C-3 beta-OH group of the saponin. The saponin is covalently coupled to an amino acid residue of a fourth proteinaceous molecule via the glucuronic acid functional group on the carbohydrate substituent at the C-3 beta-OH group of the saponin, preferably via at least one linker. The amino acid residue is preferably selected from cysteine ​​and lysine.

[0321] An embodiment of the fifth series of aspects and embodiments of the invention is a therapeutic combination for use according to the invention, or a fourth pharmaceutical composition for use according to the invention, wherein the glucuronic acid functionality on the carbohydrate substituent at the C-3 beta-OH group of at least one saponin is covalently attached to the linker 1-[bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium 3-oxide hexafluorophosphate, which is covalently attached by an amide bond to an amine group on a fourth proteinaceous molecule, for example to a lysine or N-terminal amine group of the fourth proteinaceous molecule.

[0322] An embodiment of the fifth set of aspects and embodiments of the invention is a therapeutic combination for use according to the invention, or a fourth pharmaceutical composition for use according to the invention, wherein the epitope is an epitope on a tumor cell receptor, preferably a tumor cell specific epitope, and the receptor is preferably a tumor cell specific receptor, more preferably CD71, CA125, EpCAM (17-1A), CD52, CEA, CD44v6, FAP, EGF-IR, integrin, syndecan-1, vascular integrin alpha-V beta-3, HER2, EGFR, CD20, CD22, folate receptor 1, CD146, CD56, CD1 9, CD138, CD27L receptor, PSMA, CanAg, integrin-alpha V, CA6, CD33, mesothelin, Cripto, CD3, CD30, CD239, CD70, CD123, CD352, DLL3, CD25, ephrin A4, MUC1, Trop2, CEACAM5, CEACAM6, HER3, CD74, PTK7, Notch3, FGF2, C4.4A, FLT3, CD38, FGFR3, CD7, PD-L1, CTLA4, CD52, PDGFRA, VEGFR1, VEGFR2, most preferably CD71, HER2, and EGFR.

[0323] An embodiment of the fifth set of aspects and embodiments of the invention is a therapeutic combination for use according to the invention, or a fourth pharmaceutical composition for use according to the invention, wherein the tumor cell receptor, preferably a tumor cell-specific receptor, is internalized by the tumor cell after binding to the fourth proteinaceous molecule of the invention and / or after binding to the fifth proteinaceous molecule of the invention. Preferably, binding of the fourth proteinaceous molecule and / or the fifth proteinaceous molecule to the tumor cell receptor, e.g., the tumor cell-specific receptor, is followed by internalization of the complex of the fourth proteinaceous molecule and the tumor cell receptor and the complex of the fifth proteinaceous molecule and the tumor cell receptor, mediated by the tumor cell receptor, e.g., via endocytosis.

[0324] An embodiment of the fifth set of aspects and embodiments of the invention is a therapeutic combination for use according to the invention, or a fourth pharmaceutical composition for use according to the invention, wherein the fourth proteinaceous molecule and the binding site of the fifth proteinaceous molecule comprise at least one of a monoclonal antibody, or a cell surface molecule binding domain and / or fragment thereof, preferably selected from the group consisting of cetuximab, daratumumab, gemtuzumab, trastumumab, panitumumab, brentuximab, inotuzumab, comprising or consisting of any one of moxetumomab, polatuzumab, obinutuzumab, OKT-9 anti-CD71 monoclonal antibody of the IgG type, pertuzumab, rituximab, ofatumumab, herceptin, alemtuzumab, pinatuzumab, OKT-10 anti-CD38 monoclonal antibody, an antibody of Table A2 or Table A3 or Table A4, preferably cetuximab or trastuzumab or OKT-9, or at least one cell surface molecule-binding fragment or domain thereof, with the proviso that the fourth binding site is the same as the fifth binding site.

[0325] An embodiment of the fifth series of aspects and embodiments of the present invention is a therapeutic combination for use according to the present invention, wherein the effector moiety comprised by the fifth proteinaceous molecule comprises or consists of any one or more of an oligonucleotide, a nucleic acid, and a xenonucleic acid, preferably a vector, a gene, a transgene inducing cell suicide, deoxyribonucleic acid (DNA), ribonucleic acid (RNA), an antisense oligonucleotide (ASO, AON), a short interfering RNA (siRNA), a microRNA (miRNA), a DNA aptamer, an RNA aptamer, an mRNA, a minicircle DNA, a A, peptide nucleic acid (PNA), phosphoramidate morpholino oligomer (PMO), locked nucleic acid (LNA), bridged nucleic acid (BNA), 2'-deoxy-2'-fluoroarabinonucleic acid (FANA), 2'-O-methoxyethyl-RNA (MOE), 2'-O,4'-aminoethylene-bridged nucleic acid, 3'-fluorohexitol nucleic acid (FHNA), plasmid, glycol nucleic acid (GNA), and threose nucleic acid (TNA), or derivatives thereof, more preferably BNA, for example, BNA for silencing HSP27 protein expression.

[0326] An embodiment of the fifth series of aspects and embodiments of the invention is a therapeutic combination for use according to the invention, wherein the effector moiety comprised by the fifth proteinaceous molecule comprises or consists of at least one proteinaceous molecule, preferably selected from any one or more of peptides, proteins, enzymes such as urease and Cre recombinase, proteinaceous toxins, ribosome-inactivating proteins, protein toxins selected from Table A5, and / or bacterial toxins, plant toxins, more preferably viral toxins such as apoptin; bacterial toxins such as Shiga toxins, Shiga-like toxins, Pseudomonas The protein toxin is selected from any one or more of: P. aeruginosa exotoxin (PE), or exotoxin A of PE, full-length or truncated diphtheria toxin (DT), cholera toxin; fungal toxins, such as alpha-sarcin; plant toxins, including ribosome-inactivating proteins and type 2 ribosome-inactivating protein A chains, such as giantin-30 or giantin-32, saporin-S3 or saporin-S6, bouganin or the deimmunized derivative of bouganin, debouganin, Shiga-like toxin A, pokeweed antiviral protein, ricin, ricin A chain, modeccin, modeccin A chain, abrin, abrin A chain, volkensin, volkensin A chain, viscumin, viscumin A chain; or animal or human toxins, such as frog RNase, or granzyme B or angiogenin from humans, or any fragment or derivative thereof. Preferably, the protein toxin is giantin and / or saporin.

[0327] An embodiment of the fifth series of aspects and embodiments of the invention is a therapeutic combination for use according to the invention, wherein the effector moiety comprised by the fifth proteinaceous molecule comprises or consists of at least one payload, preferably any one or more of a ribosome-targeting toxin, an elongation factor-targeting toxin, a tubulin-targeting toxin, a DNA-targeting toxin, and an RNA-targeting toxin, more preferably emtansine, pasudotox, maytansinoid derivative DM1, maytansinoid derivative DM4, monomethyl auristatin E (MMAE, vedotin), monomethyl auristatin F (MM AF, mafodotin), calicheamicin, N-acetyl-γ-calicheamicin, pyrrolobenzodiazepine (PBD) dimer, benzodiazepine, CC-1065 analog, duocarmycin, doxorubicin, paclitaxel, docetaxel, cisplatin, cyclophosphamide, etoposide, docetaxel, 5-fluorouracil (5-FU), mitoxantrone, tubulysin, indolinobenzodiazepine, AZ13599185, cryptophycin, rhizoxin, methotrexate, anthracyclines, camptothecin analog, SN-38, DX-8951f, exatecan mesylate, Pseudomonas The steroid inhibitor is selected from one or more of a truncated form of P. aeruginosa exotoxin (PE38), a duocarmycin derivative, amanitin, α-amanitin, splicostatin, tylanstatin, ozogamicin, tesirin, amberstatin 269, and soravtansine, or a derivative thereof.

[0328] An embodiment of the fifth set of aspects and embodiments of the invention is a therapeutic combination for use according to the invention, wherein the fifth proteinaceous molecule comprises or consists of any one of gemtuzumab ozogamicin, brentuximab vedotin, trastuzumab emtansine, inotuzumab ozogamicin, moxetumomab pasudotox, and polatuzumab vedotin, and the antibody-drug conjugates of Table A2 and Table A3, or at least one tumor cell receptor-binding domain thereof, and / or at least one tumor cell receptor-binding fragment thereof, wherein said domain(s) or fragment(s) comprise an effector moiety and are preferably tumor cell-specific receptor-binding domain(s) and / or tumor cell-specific receptor-binding fragment(s).

[0329] An embodiment of the fifth series of aspects and embodiments of the invention is a therapeutic combination for use according to the invention, wherein the fourth proteinaceous molecule comprises more than one covalently attached saponin, preferably 2, 3, 4, 5, 6, 8, 10, 16, 32, 64, 128, or 1-100 saponins, or any number therebetween, for example 7, 9, 12 saponins.

[0330] One embodiment of the fifth series of aspects and embodiments of the invention is a therapeutic combination for use according to the invention, wherein at least one saponin is covalently attached directly to an amino acid residue, preferably a cysteine ​​and / or a lysine, of the fourth proteinaceous molecule, and / or via at least one linker and / or via at least one cleavable linker and / or via at least one oligomeric or polymeric backbone, preferably 1 to 8 such backbones or 2 to 4 such backbones, wherein at least one backbone is optionally dendron-based and wherein 1 to 32 saponins are covalently attached to the at least one backbone, for example 2, 3, 4, 5, 6, 8, 10, 16, 32 saponins, or any number in between, for example 7, 9, 12 saponins.

[0331] An embodiment of the fifth series of aspects and embodiments of the invention is a therapeutic agent combination for use according to the invention, wherein the cleavable linker is susceptible to cleavage under acidic, reductive, enzymatic, or photoinduced conditions. Preferably, the cleavable linker comprises a cleavable bond selected from hydrazone and hydrazide bonds that are susceptible to cleavage under acidic conditions, and / or bonds that are susceptible to proteolysis, e.g., proteolysis by cathepsin B, and / or bonds that are susceptible to cleavage under reductive conditions, e.g., disulfide bonds.

[0332] An embodiment of the fifth series of aspects and embodiments of the invention is a therapeutic combination for use according to the invention, wherein the cleavable linker undergoes cleavage in vivo under acidic conditions present in the endosomes and / or lysosomes of mammalian cells, preferably human cells, preferably at a pH between 4.0 and 6.5, more preferably at a pH ≦ 5.5.

[0333] An embodiment of the fifth series of aspects and embodiments of the invention is a therapeutic combination for use according to the invention, wherein the oligomeric or polymeric backbone comprises a polymeric or oligomeric structure and comprises at least one chemical group, at least one chemical group for covalent coupling of the backbone to an amino acid residue of said fourth proteinaceous molecule.

[0334] An embodiment of the fifth series of aspects and embodiments of the invention is a therapeutic combination for use according to the invention, wherein at least one saponin is covalently attached to a backbone polymeric or oligomeric structure via a cleavable linker according to the invention.

[0335] An embodiment of the fifth series of aspects and embodiments of the invention is a therapeutic combination for use according to the invention, wherein at least one saponin is covalently attached to a backbone polymeric or oligomeric structure via any one or more of an imine bond, a hydrazone bond, a hydrazide bond, an oxime bond, a 1,3-dioxolane bond, a disulfide bond, a thio-ether bond, an amide bond, a peptide bond or an ester bond, preferably via at least one linker.

[0336] One embodiment of the fifth series of aspects and embodiments of the present invention is a therapeutic combination for use according to the invention, wherein at least one saponin comprises an aldehyde functionality at position C-23 and, optionally, a glucuronic acid functionality on the carbohydrate substituent at the C-3 beta-OH group of at least one saponin, wherein the aldehyde functionality is involved in covalent attachment to a backbone polymeric or oligomeric structure, and / or, when present, the glucuronic acid functionality is involved in covalent attachment to a backbone polymeric or oligomeric structure, either via a direct bond or via at least one linker.

[0337] One embodiment of the fifth series of aspects and embodiments of the invention is a therapeutic combination for use according to the invention, wherein the aldehyde functionality at position C-23 of at least one saponin is covalently coupled to a linker N-ε-maleimidocaproic acid hydrazide, which is covalently coupled to a sulfhydryl group, such as a cysteine ​​sulfhydryl group, on a backbone polymeric or oligomeric structure via a thio-ether bond.

[0338] An embodiment of the fifth series of aspects and embodiments of the present invention is a therapeutic combination for use according to the invention, wherein the glucuronic acid functionality on the carbohydrate substituent at the C-3 beta-OH group of at least one saponin is covalently coupled to a linker 1-[bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium 3-oxide hexafluorophosphate, which is covalently coupled via an amide bond to an amine group on a backbone polymeric or oligomeric structure, for example a lysine or N-terminal amine group of a proteinaceous molecule.

[0339] An embodiment of the fifth series of aspects and embodiments of the present invention is a therapeutic combination for use according to the present invention, wherein at least one chemical group of the backbone for covalent coupling of the oligomeric or polymeric backbone to an amino acid residue of said fourth proteinaceous molecule is a click chemistry group, preferably selected from tetrazine, azide, alkene or alkyne, or cyclic derivatives of these groups. More preferably, the click chemistry group is azide.

[0340] An embodiment of the fifth series of aspects and embodiments of the invention is a therapeutic combination for use according to the invention, wherein the backbone polymeric or oligomeric structure comprises a linear, branched and / or cyclic polymer, oligomer, dendrimer, dendron, dendronized polymer, dendronized oligomer, DNA, polypeptide, polylysine, polyethylene glycol, or an assembly of these polymeric or oligomeric structures, which assembly is preferably assembled by covalent crosslinking.

[0341] An embodiment of the fifth set of aspects and embodiments of the invention is a therapeutic combination for use according to the invention, wherein the fourth pharmaceutical composition and the fifth pharmaceutical composition are administered to a patient in need thereof.

[0342] An aspect of the invention of the fifth series of aspects and embodiments of the invention relates to a pharmaceutical composition of the fourth invention, which further comprises a fifth proteinaceous molecule of the invention.

[0343] An aspect of the invention of the fifth series of aspects and embodiments of the invention relates to a pharmaceutical composition of the fourth invention further comprising a proteinaceous molecule of the fifth invention for use as a medicament.

[0344] An aspect of the invention of the fifth series of aspects and embodiments of the invention relates to a pharmaceutical composition of the fourth invention further comprising a fifth proteinaceous molecule of the invention for use in the treatment or prevention of cancer in a patient in need thereof.

[0345] One aspect of the present invention in the sixth series of aspects and embodiments of the present invention relates to a therapeutic combination for use as a pharmaceutical, the therapeutic combination comprising: (a) a sixth pharmaceutical composition comprising a sixth proteinaceous molecule comprising a sixth binding site for binding to a sixth cell surface molecule and at least one saponin covalently attached to said sixth proteinaceous molecule, preferably covalently attached to an amino acid residue of said sixth proteinaceous molecule (the sixth pharmaceutical composition optionally further comprising a pharmaceutically acceptable excipient); and (b) a seventh pharmaceutical composition comprising a seventh proteinaceous molecule, preferably different from the...

Claims

1. A therapeutic drug molecule having the chemical structure of Compound I: A1 ｍ ((-L9 ｗ )((-L1 ｑ -B1 ｎ ) ｕ ((-L2 ｒ -L3 ｓ )(-L4 ｖ -C) ｐ ) ｔ ) ｘ (Compound I) wherein A1 is a first ligand when B1 is a first effector moiety, or A1 is a first effector moiety when B1 is a first ligand; C is a saponin; When A1 is a first ligand and B1 is a first effector moiety, then m=1; when A1 is a first effector moiety and B1 is a first ligand, m=1-32; When B1 is the first ligand and A1 is the first effector moiety, or when A1 is the first ligand and B1 is the first effector moiety, n=1; p=any of 1 to 128; L1 is at least one linker for covalently coupling two chemical groups; L2 is at least one linker for covalently coupling two chemical groups; L3 is at least one oligomeric or polymeric backbone for covalently coupling two chemical groups; L4 is at least one linker for covalently coupling two chemical groups; L9 is a trifunctional linker for covalently coupling three chemical groups; q=1; r=0 or 1; s=0 or 1; When s=0, t=1 or 2, when s=1, t=1 to 16; When A1 is a first ligand and B1 is a first effector moiety, then u=any of 1-32; or if A1 is a first effector moiety and B1 is a first ligand, then u=1; v=1; w=1 or 0; x=1 to 16.

2. The therapeutic drug molecule of claim 1, wherein the first ligand A1 or B1 comprises or consists of an immunoglobulin, a binding domain or binding fragment of an immunoglobulin, an antibody, an IgG, a molecule comprising or consisting of a Vhh domain or a Vh domain, a Fab, a scFv, an Fv, a dAb, an F(ab)2, an Fcab fragment, at least one non-proteinaceous ligand, and / or at least one proteinaceous ligand, and is a ligand for binding to a cell surface molecule, EGF or a cytokine.

3. 3. The therapeutic drug molecule of claim 1 or 2, wherein the first ligand A1 or B1 is selected from proteins, polypeptides, peptides, small molecules, interferons, transferrin, lectins, epidermal growth factor (EGF) and EGF-like domains, gastrin releasing peptide (GRP), platelet derived growth factor (PDGF), transforming growth factor (TGF), vaccinia growth factor (VGF), insulin and insulin-like growth factors, other suitable hormones, thyrotropin releasing hormone (TRH), melanocyte stimulating hormone (MSH), steroid hormones, somatostatin, lymphokines, colony stimulating factors, bombesin, gastrin, Arg-Gly-Asp or RGD, aptamers, small molecules, vitamins, carbohydrates.

4. The first ligand A1 or B1 binds between an epitope of a tumor cell or a tumor cell specific epitope and a tumor cell receptor or a tumor cell specific receptor, or binds to CD71, CA125, EpCAM (17-1A), CD52, CEA, CD44v6, FAP, EGF-IR, integrin, syndecan-1, vascular integrin alpha-V beta-3, HER2, EGFR, CD20, CD22, folate receptor 1, CD146, CD56, CD19, CD138, CD27L receptor, PSMA, CanAg , integrin alpha V, CA6, CD33, mesothelin, Crypto, CD3, CD30, CD239, CD70, CD123, CD352, DLL3, CD25, ephrin A4, MUC1, Trop2, CEACAM5, CEACAM6, HER3, CD74, PTK7, Notch3, FGF2, C4.4A, FLT3, CD38, FGFR3, CD7, PD-L1, CTLA4, CD52, PDGFRA, VEGFR1, VEGFR2.

5. The first ligand A1 or B1 is selected from the group consisting of cetuximab, daratumumab, gemtuzumab, trastuzumab, panitumab, brentuximab, inotuzumab, moxetumomab, polatuzumab, obinutuzumab, OKT-9 anti-CD71 monoclonal antibody, pertuzumab, rituximab, ofatumumab, alemtuzumab, pinatuzumab, OKT-10 anti-CD38 monoclonal antibody, tositumomab, ibritumomab, oregomab. , edrecolomab, nimotuzumab, huMy9-6 anti-CD33 monoclonal antibody, etaracizumab, epratuzumab, labetuzumab, bivatuzumab, sibrotuzumab, huB4 anti-CD19 monoclonal antibody, huC242 anti-CanAg monoclonal antibody, huN901 anti-CD56 monoclonal antibody, DS6 anti-CA6 monoclonal antibody, cixutuzumab, 3B7 anti-IGF-IR monoclonal antibody, CNTO 95 anti-integrin monoclonal antibody, B-B4 anti-syndecan-1 monoclonal antibody, or at least one tumor cell receptor binding domain and / or at least one tumor cell receptor binding fragment thereof, or the antibody drug conjugate comprises or consists of Gemtuzumab ozogamicin, Brentuximab vedotin, Trastuzumab emtansine, Inotuzumab ozogamicin, Moxetumomab pasudotox, Polatuzumab vedotin and Affilutin, IMGN-779, Neuradiab, AGS-67E, ASG-15ME, vandortuzumab vedotin, CDX-014, AGS-16M18, vorsetuzumab mafodotin, denintuzumab mafodotin, SGN-CD70A, RG-7636, SC-006, MM-310, PF-06647263, PF-06263507, PF-06650808, XMT-1522, AMG-595, pinatuzumab vedotin, cantuzumab ravtansine, AVE-9633, BIWI-1, RG-7882, ASG-5ME, DCDS-0780A, SC-004, RG-7600, sofituzumabvedotin, IMGN-289, SAR-428926, SGNCD-19B, SGNCD-123A, SGNCD-352A, RG-7841, IMGN-388, lorvotuzumab mertansine, BAY-794620, RG-7598, Oncolysin B, ADCT-502, AMG-172, ImmuRAIT-LL2, indusatumab vedotin, clivatuzumab tetraxetan, depatuxizumab mafodotin, vadastuximab talirine, MLN-2704, coltuximab ravtansine, lifastuzumab vedotin, Bismab-A, Avicidin, SGN-15, ASP-6183, SAR-566658, Oncolysin S, glembatumumab vedotin, MM-302, HTI-1511, ZW-33, HuMax-CD74-ADC, sacituzumab govitecan, ibritumomab tiuxetan, rovalpituzumab The therapeutic molecule of any one of claims 1 to 4, which is tesirine, or at least one tumor cell receptor binding domain and / or at least one tumor cell receptor binding fragment thereof, which is a tumor cell specific receptor binding domain(s) and / or a tumor cell specific receptor binding fragment(s).

6. The therapeutic molecule of any one of claims 1 to 5, wherein the first ligand A1 is internalized by a tumor cell following binding of the first ligand to its binding partner on the tumor cell and / or binding of the first ligand to the tumor cell is followed by internalization of the complex of the first ligand and its binding partner on the tumor cell by a tumor cell receptor, e.g. via endocytosis.

7. 7. The therapeutic molecule of any one of claims 1 to 6, wherein the first effector moiety A1 or B comprises or consists of at least one of any one or more of an oligonucleotide, a nucleic acid, and a xenonucleic acid, or is selected from any one or more of a vector, a gene, a transgene inducing cell suicide, deoxyribonucleic acid (DNA), ribonucleic acid (RNA), an antisense oligonucleotide (ASO, AON), a short interfering RNA (siRNA), a microRNA (miRNA), a DNA aptamer, an RNA aptamer, an mRNA, a minicircle DNA, a peptide nucleic acid (PNA), a phosphoramidate morpholino oligomer (PMO), a locked nucleic acid (LNA), a bridged nucleic acid (BNA), a 2'-deoxy-2'-fluoroarabinonucleic acid (FANA), a 2'-O-methoxyethyl-RNA (MOE), a 2'-O,4'-aminoethylene bridged nucleic acid, a 3'-fluorohexitol nucleic acid (FHNA), a plasmid, a glycol nucleic acid (GNA), and a threose nucleic acid (TNA), or a derivative thereof.

8. The therapeutic molecules include gemtuzumab ozogamicin, brentuximab vedotin, trastuzumab emtansine, inotuzumab ozogamicin, moxetumomab pasudotox, polatuzumab vedotin, and affilin, ADCT-502, AGS-16M18, AGS-67E, AMG-172, AMG-595, ASG-15ME, ASG-5ME, ASP-6183, AVE-9633, avicidin, BAY-794620, bisumab-A, BIWI-1, cantuzumab ravtansine. Shin, CDX-014, cortuximab ravtansine, DCDS-0780A, denintuzumab mafodotin, HTI-1511, HuMax-CD74-ADC, IMGN-289, IMGN-388, IMGN-779, ImmuRAIT-LL2, industamab vedotin, rifastamab vedotin, lorvotuzumab mertansine, MLN-2704, MM-310, Oncolysin S, PF-06263507, PF-06647263, PF-06650808, pinotuzumab buvedotin, RG-7598, RG-7600, RG-7636, RG-7841, RG-7882, sacituzumab govitecan, SAR-428926, SAR-566658, SC-004, SC-006, SGN-15, SGNCD-123A, SGNCD-19B, SGNCD-352A, SGN-CD70A, sofituzumab vedotin, bundurumab vedotin, borsetuzumab mafodotin, XMT-1522, ZW-33, trastuzumab emtansine, MM-302, The therapeutic molecule according to any one of claims 1 to 7, comprising or consisting of any one of ibritumomab tiuxetan, inotuzumab ozogamicin, rovalpituzumab tesirin, neuradiab, clivatuzumab tetraxetan, depatuximab mafodotin, vadastoximab butarilin, glembatuzumab vedotin, and oncolysin B, and comprising or consisting of at least one tumor cell specific receptor binding domain and / or tumor cell specific receptor binding fragment.

9. 9. The therapeutic molecule of any one of claims 1 to 8, wherein Saponin C is a triterpenoid saponin or a bisdesmosidic triterpene saponin belonging to the type of 12,13-dehydrooleanane having an aldehyde functionality at position C-23 and optionally a glucuronic acid functionality in the carbohydrate substituent of the C-3 beta-OH group of the saponin, and / or a saponin isolated from Gypsophila species and / or Saponaria species and / or Agrostenma species and / or Quillaja species, Quillaja saponaria.

10. 10. The therapeutic agent molecule of any one of claims 1 to 9, wherein Saponin C is a triterpenoid saponin and / or a bisdesmosidic triterpene saponin belonging to the 12,13-dehydrooleanane type and having an aldehyde functional group at the C-23 position and selected from the group consisting of saponins having a quillajaic acid aglycone core and saponins having a gypsogenin aglycone core, said saponin comprising a first branched carbohydrate side chain at the C-3 position, said first branched carbohydrate side chain comprising a glucuronic acid functional group, and said saponin comprising an ester group comprising a second branched carbohydrate side chain comprising a deoxy carbohydrate at the C-28 position.

11. 11. The therapeutic molecule of any one of claims 1 to 10, wherein the saponin is a triterpenoid saponin and / or a bisdesmosidic triterpene saponin, belonging to the 12,13-dehydrooleanane type, has an aldehyde functional group at the C-23 position, and is selected from the group consisting of saponins having a quillaic acid aglycone core and saponins having a gypsogenin aglycone core, and / or wherein said at least one saponin comprises a first branched carbohydrate side chain comprising a glucuronic acid functional group at the C-3 position and said saponin comprises an ester group comprising a second branched carbohydrate side chain comprising a deoxy carbohydrate at the C-28 position.

12. Saponin C may be a single specific saponin or a mixture of two or more different saponins, such as one or more of the saponins in Table A1 or Scheme I, SO1861, SA1657, GE1741, SA1641, QS-21, QS-21A, QS-21 A-api, QS-21 A-xyl, QS-21B, QS-21 B-api, QS-21 B-xyl, QS-7-xyl, QS-7-api, QS-17-api, QS-17-xyl, QS1861, QS1862, Quillaja saponin, Saponinum albumensis, album), QS-18, Quil-A, Gyp1, gypsoside A, AG1, AG2, SO1542, SO1584, SO1658, SO1674, SO1832, or any of their stereoisomers and / or any combination thereof, or the saponin is SO1861 and / or GE1741 and / or SA1641 and / or QS-21 and / or a quillaric acid aglycone core, a Gal-(1→2)-[Xyl-(1→3)]-GlcA carbohydrate substituent at the C-3 beta-OH group, and a Glc-(1→3)-Xyl-(1→4)-Rha-( 12. The therapeutic molecule of any one of claims 1 to 11, which is a saponin having a 1→2)-[Xyl-(1→3)-4-OAc-Qui-(1→4)]-Fuc carbohydrate substituent; and / or 3-O-beta-D-galactopyranosyl-(1→2)-[beta-D-xylopyranosyl-(1→3)]-beta-D-glucuronopyranosylquilayic acid, 28-O-beta-D-glucopyranosyl-(1→3)-beta-D-xylopyranosyl-(1→4)-alpha-L-rhamnopyranosyl-(1→2)-[beta-D-xylopyranosyl-(1→3)-4-OAc-beta-D-quinovopyranosyl-(1→4)]-beta-D-fucopyranoside.

13. Saponin C is a bisdesmosidic saponin having a molecular mass of at least 1.500 Daltons and comprising an oleanane-type triterpene containing an aldehyde group at the C-23 position and optionally a hydroxyl group at the C-16 position, a first branched carbohydrate side chain at the C-3 position, which first branched carbohydrate side chain optionally contains glucuronic acid, the saponin contains an ester group at the C-28 position and optionally a second branched carbohydrate side chain, which second branched carbohydrate chain contains at least four carbohydrate units, optionally at least one acetyl residue, two acetyl residues, and / or at least one deoxy carbohydrate; and / or quinovose, and / or glucose, and / or 4-methoxycinnamic acid, and / or optionally 5-O-[5-O-Ara / Api-3,5-dihydroxy-6-methyl-octanoyl]-3,5-dihydroxy-6-methyl-octanoic acid, and / or optionally 5-O-[5-O-Rha-(1→2)-Ara / Api-3,5-dihydroxy-6-methyl-octanoyl]-3,5-dihydroxy-6-methyl-octanoic acid, linked to the carbohydrate via an ester bond, or at least one saponin is selected from the group consisting of QS-21, QS-21A, QS-21B, QS-21C, QS-21D, QS-21E, QS-21F, QS-21G, QS-21H ... The therapeutic molecule of any one of claims 1 to 12, which is one or more of: A-api, QS-21 A-xyl, QS-21B, QS-21 B-api, QS-21 B-xyl, QS-7-xyl, QS-7-api, QS-17-api, QS-17-xyl, QS-18, QS-1861, protonated QS1861 (QS1862), and Quil-A.

14. The therapeutic drug molecule of any one of claims 1 to 13, wherein saponin C is a bisdesmosidic triterpene saponin belonging to the type of 12,13-dehydroanane having an aldehyde functional group at position C-23, and saponin C is covalently coupled to an amino acid residue of the first ligand A1 or B1 and / or the first effector moiety B1 or A1 via the aldehyde functional group on saponin C or said aldehyde functional group at position C-23, optionally via a linker L2, L4, L9 or via a cleavable linker L2, L4, L9, or the amino acid residue is selected from cysteine ​​and lysine.

15. 15. The therapeutic drug molecule of any one of claims 1 to 14, wherein saponin C is a bisdesmosidic triterpene saponin belonging to the type of 12,13-dehydroanane having an aldehyde function at position C-23, and wherein the aldehyde function at position C-23 of at least one saponin is covalently coupled to a linker N-ε-maleimidocaproic acid hydrazide, which is covalently coupled via a thio-ether bond to a sulfhydryl group on the first ligand A1 or B1 and / or the first effector moiety B1 or A1, or to a sulfhydryl group of a cysteine.

16. 16. The therapeutic drug molecule of any one of claims 1 to 15, wherein Saponin C is a bisdesmosidic triterpene saponin belonging to the 12,13-dehydrooleanane type having an aldehyde functionality at position C-23 and comprising a glucuronic acid functionality on a carbohydrate substituent at the C-3 beta-OH group of the saponin, and Saponin C is covalently coupled to an amino acid residue of the first ligand A1 or B1 and / or the first effector moiety B1 or A1 via the glucuronic acid functionality on Saponin C, if present, or via a linker L2, L4, L9, or the amino acid residue is selected from cysteine ​​and lysine, or the amino acid residue is lysine.

17. 17. The therapeutic drug molecule of any one of claims 1 to 16, wherein saponin C is a bisdesmosidic triterpene saponin belonging to the type of 12,13-dehydrooleanane having an aldehyde functional group at position C-23 and comprising a glucuronic acid functional group on a carbohydrate substituent at the C-3 beta-OH group of the saponin, wherein at least one glucuronic acid functional group on a carbohydrate substituent at the C-3 beta-OH group of the saponin is covalently coupled to a linker 1-(bis(dimethylamino)methylene)-1H-1,2,3-triazolo[4,5-b]pyridinium 3-fluorophosphate, which is covalently coupled via an amide bond to an amine group of the first ligand A1 or B1 and / or the first effector moiety B1 or A1, or to a lysine or N-terminal amine group of the first ligand A1 or B1 and / or the first effector moiety B1 or A1.

18. 18. The therapeutic molecule of any one of claims 1 to 17, wherein the first ligand A1 or B1 and / or the first effector moiety B1 or A1 comprises one or more than one covalently attached saponin C, or 2, 3, 4, 5, 6, 8, 10, 16, 32, 64, 128, or 1-100 saponins, or any number in between, or 7, 9, 12 saponins.

19. The first ligand A1 or B1 and / or the first effector moiety B1 or A1 comprises one or more covalently bound saponin C, wherein the saponin C(s) is / are directly covalently bound to an amino acid of the first ligand A1 or B1 and / or the first effector moiety B1 or A1 (wherein r, s, v, e, f, and i are 0), optionally to a cysteine ​​and / or a lysine, and / or via at least one linker L2, L4, L9, or via at least one cleavable linker.

19. The therapeutic molecule of any one of claims 1 to 18, wherein the saponins are covalently linked via possible linkers L2, L4, L9 and / or via at least one oligomeric or polymeric backbone L3, or 1 to 8 such backbones or 2 to 4 such backbones, at least one backbone optionally being based on a dendron, and wherein 1 to 32 saponins, or 2, 3, 4, 5, 6, 8, 10, 16, 32 saponins, or any number of saponins in between, or 7, 9, 12 saponins are covalently linked to the at least one backbone.

20. The therapeutic drug molecule of claim 14, wherein the cleavable linkers L2, L4, L9 are cleaved under acidic, reductive, enzymatic or photoinduced conditions, and optionally the cleavable linkers comprise a hydrazone or hydrazide bond that is cleaved under acidic conditions when bound to saponin, and / or a bond that is susceptible to proteolysis, e.g., proteolysis by cathepsin B, when bound to saponin, and / or the cleavable linkers comprise a disulfide bond that is susceptible to cleavage under reductive conditions.

21. The therapeutic drug molecule of any one of claims 14 to 20, wherein the cleavable linkers L2, L4, L9 undergo cleavage in vivo under acidic conditions present in endosomes and / or lysosomes of mammalian or human cells or at a pH of 4.0 to 6.

5.

22. 22. The therapeutic drug molecule of any one of claims 1 to 21, wherein the polymer or oligomeric backbone L3 comprises a polymer or oligomeric structure and comprises chemical groups for covalent coupling of the polymer or oligomeric backbone L3 to amino acid residues of the chemical group, the first ligand and / or the first effector moiety.

23. 23. The therapeutic drug molecule of any one of claims 1 to 22, wherein at least one saponin is covalently attached to the polymeric or oligomeric structure of the backbone L3 via a cleavable linker L4 as described in any one of claims 18 to 22.

24. 24. The therapeutic drug molecule of any one of claims 18 to 23, wherein the chemical group of the polymer or oligomer backbone L3 for covalently coupling the backbone to an amino acid residue of the first ligand and / or the first effector moiety is a click chemistry group or is selected from a tetrazine, an azide, an alkene or an alkyne or cyclic derivatives of these groups, or the click chemistry group is an azide.

25. 25. The therapeutic drug molecule of any one of claims 18-24, wherein the at least one saponin is covalently attached to the first ligand and / or first effector moiety either directly or via at least one linker, for example a bifunctional linker based on N-ε-maleimidocaproic acid hydrazide and / or based on 1-[bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium 3-oxide hexafluorophosphate, or a trifunctional linker L9 when w=1 and / or a trifunctional linker L10 when j=1, e.g. a trifunctional linker of Scheme II.

26. The therapeutic drug molecule of claim 25, wherein the trifunctional linker L9 when w=1 comprises a second chemical group to which at least one saponin is covalently attached, a third chemical group for covalently attaching to a first ligand, and a first chemical group for covalently attaching to at least one first effector moiety.

27. 27. The therapeutic molecule of any one of claims 1 to 26, wherein the at least one saponin is covalently attached to the first ligand and / or the first effector moiety via at least one linker comprising a trifunctional linker L9, where j=1, to which both the first ligand and the at least one first effector moiety are attached.

28. The therapeutic drug molecule of any one of claims 18 to 27, wherein the polymeric or oligomeric structure of backbone L3 comprises a linear, branched, and / or cyclic polymer, oligomer, dendrimer, dendron, dendronized polymer, dendronized oligomer, DNA, polypeptide, polylysine, polyethylene glycol, or an assembly of these polymeric or oligomeric structures, which assembly is assembled by covalent crosslinking.

29. The therapeutic molecule of any one of claims 18 to 27, wherein the first ligand A1 or B1 is covalently linked to the first effector moiety B1 or A1, respectively, via at least one linker L1.

30. The therapeutic molecule of claim 1, wherein r=0, s=0, v=0, s=0, v=0, p=0, t=0, the ligand A1 is a monoclonal antibody or at least one binding fragment or domain thereof of any one of claims 3-7, m=1, the effector moiety B1 is an effector moiety of claim 7, either q=0, n=1, u=2-4 or q=1, n=2-4, u=2-4, and the linker L1 is an oligomeric or polymeric scaffold L3 of any one of claims 19-28.

31. A first pharmaceutical composition comprising a therapeutic drug molecule having the chemical structure of Compound I according to any one of claims 1 to 30, the first pharmaceutical composition optionally further comprising a pharma- ceutically acceptable excipient.

32. 32. The first pharmaceutical composition of claim 31 for use as a medicament.

33. The first pharmaceutical composition according to claim 31 or 32 for use in the treatment or prevention of cancer or an autoimmune disease.

34. A first pharmaceutical composition according to any one of claims 31 to 33, wherein ligand A1 or B1 is capable of binding to a tumor cell epitope, or a tumor cell specific epitope, on a tumor cell surface molecule, or a tumor cell specific surface molecule, for use in the treatment or prevention of cancer in a patient in need thereof.

35. The first pharmaceutical composition for use according to claim 31 or 32, wherein the first pharmaceutical composition according to any one of claims 31 to 34 is administered to a patient in need thereof.

36. An antibody-drug conjugate (ADC) or antibody-oligonucleotide (AOC) or semi-finished ADC conjugate or semi-finished AOC conjugate, comprising an antibody-saponin conjugate, comprising at least one effector molecule and / or at least one saponin, having the structure C: A(-S)b(-E)c structure C, A is an antibody; S is a saponin; E is an effector molecule; b is 0-64, or 0, 1, 2, 3, 4, 8, 16, 32, 64 or any integer or fraction therebetween; c is 0-8, or 0, 1, 2, 3, 4, 6, 8 or any integer or fraction therebetween; S binds to A and / or E, E binds to A and / or S, A is selected from an anti-EGFR antibody, an anti-HER2 antibody, an anti-CD71 antibody, and / or wherein S is any one or more of saponins, triterpenoid saponins and / or bisdesmosidic triterpene saponins belonging to the type of 12,13-dehydrooleanane having an aldehyde function at the C-23 position, optionally containing a glucuronic acid function in the carbohydrate substituent of the C-3 β-OH group of the saponin, such as SO1861, GE1741, SA1641, Quil-A, QS-21, and saponins in the water-soluble saponin fraction of Quillaja saponaria; The saponin and / or effector molecule are covalently attached via at least one linker or cleavable linker, and / or at least one oligomeric or polymeric scaffold, or via linkers based on N-ε-maleimidocaproic acid hydrazide (EMCH), succinimidyl 3-(2-pyridyldithio)propionate or 3-(2-pyridyldithio)propionic acid N-hydroxysuccinimide ester (SPDP), and 1-[bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium 3-oxide hexafluorophosphate (HATU), and / or dendrons such as G4-dendrons or trifunctional linkers or the trifunctional linkers of Scheme II. An antibody-drug conjugate (ADC) or antibody-oligonucleotide (AOC) or semi-finished ADC conjugate or semi-finished AOC conjugate, wherein the antibody first binding site or the monoclonal antibody or a fragment or domain thereof is covalently linked via a functional linker-based scaffold and / or at least one lysine side chain and / or cysteine ​​side chain of the antibody first binding site or the monoclonal antibody or a fragment or domain thereof is involved in a covalent bond with the saponin and / or effector molecule and / or linker and / or cleavable linker and / or scaffold, or the saponin and / or effector molecule is covalently linked to the antibody first binding site or the monoclonal antibody, wherein the covalent bond comprises or consists of an amide bond, a hydrazone bond, a disulfide bond.

37. 37. An antibody-drug conjugate (ADC) or an antibody-oligonucleotide (AOC) or a semi-finished ADC conjugate or a semi-finished AOC conjugate according to claim 36 for use as a medicament.

38. 38. An antibody-drug conjugate (ADC) or antibody-oligonucleotide (AOC) or semi-finished ADC conjugate or semi-finished AOC conjugate according to claim 36 or 37 for use in the treatment or prevention of cancer or an autoimmune disease.

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