Camptothecin conjugate

Abstract

Antibody conjugates of camptothecin compounds are described, along with their use and preparation methods.

Description

[0001] Cross-reference to related applications

[0002] This application claims priority to U.S. Provisional Patent Application Serial No. 62 / 681,847, filed June 7, 2018, and U.S. Provisional Patent Application Serial No. 62 / 777,491, filed December 10, 2018, both of which are incorporated herein by reference in their entirety for all purposes. Background Technology

[0003] Antibodies (mAbs) have been investigated for targeted delivery of cytotoxic agents to tumor cells. While various drug classes have been evaluated for targeted delivery via antibodies, only a few have proven to be sufficiently active as antibody-drug conjugates, possessing suitable toxicity and other pharmacological properties to warrant clinical development. One drug class of interest is camptothecin.

[0004] The design of antibody-drug conjugates (ADCs) involves linking a cytotoxic agent to an antibody via a linker, typically involving consideration of several factors, including the presence of a conjugate stalk on the drug for linking to the linker and the linker technique for linking the drug to the antibody in a conditionally stable manner. In this category, the conjugate stalk of the parent compound is a C20 hydroxyl functional group, with the linker linked via a carbonate functional group (see, for example, Walker, MA et al., Bioorganic & Medicinal Chemistry Letters (2002) 12(2):217-219). However, carbonate functional groups are generally susceptible to hydrolytic instability, which causes premature release of the free drug into the systemic circulation, potentially leading to reduced ADC efficacy, insufficient immunospecificity of the conjugate, and increased toxicity. Therefore, there is a need in the art for camptothecin conjugates with improved stability to increase the amount of drug delivered to the desired site of action. This invention addresses those and other needs. Invention Overview

[0006] This invention specifically provides camptothecin conjugates, camptothecin-linker compounds, and camptothecin compounds, methods for preparing and using them, and intermediates thereof. The camptothecin conjugates of this invention are stable in circulation, but can cause cell death once the conjugate releases free drug near or inside tumor cells.

[0007] In one main embodiment, a camptothecin conjugate or a salt thereof having the following formula is provided:

[0008] L-(QD) p

[0009] in

[0010] L stands for ligand unit;

[0011] The subscript p is an integer from 1 to 16;

[0012] Q is a connection subunit having a formula selected from the following:

[0013] -ZA-,-ZA-RL-,-ZA-RL-Y-,-ZAS * -RL-,-ZAS * -RL-Y-,

[0014] ZAS * -W-,-ZAS * -W-RL-,-ZAB(S * )-RL-,-ZAB(S * )-W-,

[0015] -ZAB(S * -W-RL- and -ZAB(S * )-RL-Y-,

[0016] Where Z is the stretcher unit.

[0017] A stands for key or connector unit;

[0018] B stands for Parallel connector unit;

[0019] S * It is a partitioning agent;

[0020] RL stands for releaseable connector;

[0021] W stands for amino acid unit;

[0022] Y represents the spacer unit; and

[0023] D is a drug unit selected from the following:

[0024]

[0025] in

[0026] R B The derivative is selected from H, C1-C8 alkyl, C1-C8 haloalkyl, C3-C8 cycloalkyl, (C3-C8 cycloalkyl)-C1-C4 alkyl, phenyl, and phenyl-C 1- Members of C4 alkyl groups;

[0027] R C Selected from C1-C6 alkyl and C 3-Members of C6 cycloalkyl groups;

[0028] R F and R F’ Each is independently selected from -H, C1-C8 alkyl, C1-C8 hydroxyalkyl, C1-C8 aminoalkyl, (C1-C4 alkylamino)-C1-C8 alkyl, N,N-(C1-C4 hydroxyalkyl)(C1-C4 alkyl)amino-C1-C8 alkyl, N,N-bis(C1-C4 alkyl)amino-C 1- C8 alkyl, N-C1-C4 hydroxyalkyl-C1-C8 aminoalkyl, C1-C8 alkyl C(O)-, C1-C8 hydroxyalkyl-C(O)-, C1-C8 aminoalkyl C(O)-, C3-C 10 cycloalkyl, (C3-C 10 Cycloalkyl)-C1-C4 alkyl, C3-C 10 Heterocyclic alkyl groups, (C3-C 10 Members of heterocyclic alkyl-C1-C4 alkyl, phenyl, phenyl-C1-C4 alkyl, diphenyl-C1-C4 alkyl, heteroaryl, and heteroaryl-C1-C4 alkyl; or

[0029] R F and R F’ It combines with the nitrogen atoms to form a 5-, 6-, or 7-membered ring having 0 to 3 substituents selected from halogens, C1-C4 alkyl groups, -OH, -OC1-C4 alkyl groups, -NH2, and -NHC. 1- C4 alkyl and -N(C1-C4 alkyl)2; and

[0030] Where R B R C R F and R F’ The cycloalkyl, heterocycloalkyl, phenyl, and heteroaryl moieties are composed of 0 to 3 elements selected from halogens, C 1- C4 alkyl, -OH, -OC1-C4 alkyl, -NH2, -NHC 1- C4 alkyl and -N(C 1- Substituents of C4 alkyl)2; and

[0031] Where Q is -ZA-RL-, -ZA-RL-Y-, -ZAS * -RL-、-ZAB(S * -RL-、-ZAS * -RL-Y- or -ZAB(S) *)-RL-Y-, where RL is any of the releasable linkers disclosed herein, and the connection point between D and Q passes through a heteroatom of any of the hydroxyl, primary, or secondary amine functional groups present on CPT1, CPT2, CPT3, CPT4, CPT5, CPT6, or CPT7, or

[0032] Where Q is -ZA- or -ZAS * -W- or -ZAB(S * -W-, or when Q is -ZAS * -RL-、-ZAB(S * -RL-、-ZAS * -W-RL- or -ZAB(S * When RL is a releasable unit other than a glucuronide unit, the connection point between D and Q passes through the oxygen atom of the hydroxyl substituent in the lactone ring of CPT1, CPT2, CPT3, CPT4, CPT5, CPT6, or CPT7; and

[0033] The condition is that when the linkage point is a nitrogen atom of a CPT6 primary or secondary amino group, R F and R F’ At least one of them is -H,

[0034] The condition is that when D is a CPT1 with nitrogen atom linked through its primary amino group, -ZA-RL-, -ZA-RL-Y-, -ZAS * -RL-、-ZAB(S * -RL-、-ZAS * -RL-Y- and -ZAB(S * )-RL-Y- of -ZA- is not optionally succinimide-hexanoyl-β-alanyl with a hydrolyzed succinimide ring.

[0035] Other principal embodiments described above are camptothecin-linker compounds that can be used as intermediates for the preparation of camptothecin conjugates, wherein the camptothecin-linker compound consists of camptothecin and a linker unit (Q), wherein the linker unit consists of an extended subunit precursor (Z') capable of forming a covalent bond with a targeting ligand providing a ligand unit and a releasable linker (RL), the releasable linker being a glucuronide unit in some aspects where Q does not have an amino acid unit.

[0036] In another aspect, this article provides a method for treating cancer, which includes administering the camptothecin conjugate described herein to subjects in need of it.

[0037] In another aspect, this article provides a kit containing the camptothecin conjugate described herein. Attached Figure Description

[0038] Figure 1A-1B Evaluation of glucuronide-based camptothecin ADCs in an in vitro bystander activity model of multiple myeloma. A. Dose-responsive titration of anti-Ag5 camptothecin DAR8 ADC (Ag5-(67)) against MM.1R(Ag5+) cell lines. (Used by CellTitre-Globe) TM Viability was assessed. Anti-Ag5 camptothecin DAR8 ADC (Ag5-(67)) dose-response titration was performed on a 3:1 co-culture mixture of B.MM.1R and MM.1R Ag5 KO (crispr cas knockout) luc+ cell lines. Viability was assessed by Bright-Globe assay. TM Assess survivability.

[0039] Figure 2 Results of an ADC stability study in mouse plasma are presented. A. camptothecin DAR8 ADC was incubated in mouse plasma (BalbC) at 37°C. Plasma samples were collected at 6 h, 24 h, 72 h, and 7 days. ADC was isolated from plasma using IgSelect, deglycosylated with PNGase, and reduced with dithiothreitol. Heavy and light chains of the ADC were assessed by PLRP-MS to quantify drug load at each time point.

[0040] Figure 3 This image shows the mean tumor volume of a 786O renal cell carcinoma subcutaneous mouse xenograft model with anti-Ag4 camptothecin DAR4 ADC (Ag4-(67)). Animals were transplanted with 786O cells. On day 7, animals were classified into groups with a mean tumor size of 100 mm. 3 The animals were then treated with single doses of camptothecin ADC at doses of 3 and 10 mg / kg. Tumor size and vital signs were assessed in the animals during the study.

[0041] Figure 4 This image shows the mean tumor volume of a subcutaneous mouse xenograft model of Hodgkin's lymphoma treated with anti-Ag2 camptothecin DAR4 ADC (Ag2-(67)). Animals were transplanted with L540cy cells. After 7 days, animals were classified into groups with a mean tumor size of 100 mm. 3 The group was then treated with a single dose of camptothecin ADC at 3 mg / kg. Tumor size and vital signs were assessed in the animals during the study.

[0042] Figure 5The results of a bystander subcutaneous xenograft tumor model of Karpas 299 / Karpas299-BVR anaplastic large cell lymphoma treated with camptothecin ADCs are shown in the figure. The figure shows the mean tumor volume against anti-Ag2 camptothecin DAR4 ADC (Ag2-(67)) and unbound DAR4 ADC (h00-(67)). Animals were transplanted with a 1:1 mixture of Ag2(+) Karpas299 and Ag2(-) Karpas299-bentuximab-resistant (Karpas299-BVR) cells. After 8 days, the animals were classified into groups with a mean tumor size of 100 mm. 3 The animals were then treated with single doses of camptothecin ADC at doses of 3, 10, and 30 mg / kg. The dose of unbound camptothecin ADC was 30 mg / kg. Tumor size and vital signs were assessed in the animals during the study.

[0043] Figure 6 This shows the average tumor volume in a mouse xenograft model of Caki-1 renal cell carcinoma using a caki-1 subcutaneous xenograft model with camptothecin DAR4 ADC. Solid caki-1 tumors were transplanted into the animals via cannula. On day 13, the animals were classified as having an average tumor size of 100 mm. 3 The animals were then treated with single doses of camptothecin ADC at doses of 10 and 30 mg / kg. Tumor size and vital signs were assessed during the study.

[0044] Figure 7 This shows the average tumor volume in a 786O renal cell carcinoma subcutaneous mouse xenograft model using a glucuronide-camptothecin-DAR4 ADC. Animals were transplanted with 786O cells. On day 7, animals were classified as having an average tumor size of 100 mm. 3 The animals were then treated with a single dose of camptothecin ADC at 10 mg / kg. Tumor size and vital signs were assessed during the study.

[0045] Figure 8 This image shows the mean tumor volume of a subcutaneous mouse xenograft model of Hodgkin's lymphoma treated with anti-Ag2 and unbound h00 glucuronide camptothecin ADC. Animals were subcutaneously transplanted with L540cy cells. After 7 days, the animals were classified into groups with a mean tumor size of 100 mm. 3 The animals were then treated with Ag2-(58) or Ag2-(61)DAR4 camptothecin ADC at doses of 10 and 30 mg / kg, respectively, for q4dx3. The corresponding dose of the unbound h00 ADC was 30 mg / kg q4dx3. Tumor size and vital signs were assessed in the animals during the study.

[0046] Figure 9Pharmacokinetic curves of h00 mAb and h00-camptothecin ADC in Sprague-Dawley rats are shown. Rats were injected with 1 mg / kg of either the parental unbound humanized antibody (h00) or h00-(67)DAR8 camptothecin ADC. Samples from predetermined blood draws were processed, and h00 parental antibody and ADC were captured from plasma using biotin-conjugated mouse anti-human light chain κ mAb and streptavidin-coated magnetic beads. h00 antibody and ADC were quantified by ELISA using the AF647-anti-human κ assay kit.

[0047] Figure 10 Image showing the mean tumor volume of camptothecin glucuronide DAR8ADC in a Del-BVR (resistant to benzoximab, MDR+) xenograft model. Del-BVR cells were subcutaneously transplanted into the animals. Four days later, the animals were classified into groups with a mean tumor size of 100 mm. 3 The animals were divided into two groups and treated with a single dose of camptothecin DAR8 ADC. Tumor size and vital signs were assessed in the animals during the study.

[0048] Figure 11 This image shows the mean tumor volume of a Caki-1 renal cell carcinoma subcutaneous mouse xenograft model using DAR4 glucuronide-camptothecin ADC. Animals were transplanted with Caki-1 cells. On day 11, animals were classified as having a mean tumor size of 100 mm. 3 The animals were then treated with a single dose of camptothecin ADC at 10 mg / kg. Tumor size and vital signs were assessed during the study.

[0049] Figure 12 This is a graph showing the activity of the selected camptothecin ADC in a nude mouse RCC xenograft model.

[0050] Figure 13 The figures and tables illustrate the plasma stability of the selected camptothecin ADC in the mouse model.

[0051] Figure 14 Further results are shown for the subcutaneous xenograft tumor model of Karpas 299 / Karpas299-BVR anaplastic large cell lymphoma treated with camptothecin ADC.

[0052] Figure 15 The camptothecin conjugate exhibits unexpected stability, wherein the conjugation is carried out by substituting the C20 hydroxyl group of the lactone ring. Invention Details

[0054] definition

[0055] Unless otherwise stated, the following terms and expressions used herein are intended to have the following meanings. When a trade name is used herein, unless the context otherwise indicates, the trade name includes the product formulation of the trade name product, the generic name of the drug, and the active pharmaceutical ingredient.

[0056] As used herein, the term "antibody" is used in the broadest sense and specifically encompasses complete monoclonal antibodies, polyclonal antibodies, monospecific antibodies, multispecific antibodies (e.g., bispecific antibodies), and antibody fragments exhibiting desired biological activity. The natural form of an antibody is a tetramer and consists of two identical pairs of immunoglobulin chains, each pair having one light chain and one heavy chain. In each pair, the light and heavy chains have variable regions (V... L and V H Together, they are primarily responsible for binding to antigens. The variable domains of the light and heavy chains consist of a framework region interrupted by three hypervariable regions (also known as "complementarity-determining regions" or "CDRs"). The constant regions are recognized and interact with by the immune system. (See, for example, Janeway et al., 2001, Immunol. Biology, 5th ed., Garland Publishing, New York). Antibodies can be of any type (e.g., IgG, IgE, IgM, IgD, and IgA), class (e.g., IgG1, IgG2, IgG3, IgG4, IgA1, and IgA2), or subclass. Antibodies can be derived from any suitable species. In some embodiments, antibodies are human or murine. Antibodies can be, for example, human antibodies, humanized antibodies, or chimeric antibodies.

[0057] As used herein, the term "monoclonal antibody" refers to an antibody derived from a substantially homogeneous population of antibodies (i.e., the individual antibodies constituting the population are identical except for the possibility of naturally occurring mutations that may exist in small amounts). Monoclonal antibodies are highly specific, targeting a single antigenic site. The modifier "monoclonal" indicates that the antibody acquires this characteristic from a substantially homogeneous population of antibodies, and should not be construed as requiring the antibody to be produced through any specific method.

[0058] As appropriate for antibody class, a "complete antibody" is one that includes an antigen-binding variable region and a light chain constant domain (C). L ) and heavy chain constant structural domain C H 1. C H 2. C H 3 and C H 4. The constant domain can be a natural sequence constant domain (e.g., a human natural sequence constant domain) or a variant of its amino acid sequence.

[0059] An "antibody fragment" comprises a portion of a complete antibody, including its antigen-binding or variable region. Examples of antibody fragments include Fab, Fab', F(ab')2, and Fv fragments, double-chain antibodies, triple-chain antibodies, quadruple-chain antibodies, linear antibodies, single-chain antibody molecules, scFv, scFv-Fc, multispecific antibody fragments formed from one or more antibody fragments, one or more fragments generated from a Fab expression library, or an epitope-binding fragment of any of the above that binds immune-specifically to a target antigen (e.g., cancer cell antigen, viral antigen, or microbial antigen).

[0060] An "antigen" is an entity that an antibody specifically binds to.

[0061] The terms "specific binding" and "specifically binding" refer to the ability of an antibody or antibody derivative to bind to the corresponding epitope of its target antigen in a highly selective manner, without binding to a variety of other antigens. Typically, the antibody or antibody derivative binds at least about 1 x 10-1 epitopes. -7 M, preferably 10 -8 M to 10 -9 M, 10 -10 M, 10 -11 M or 10 -12 M binds with its affinity and binds to the predetermined antigen with an affinity at least twice as great as that it binds to non-specific antigens other than the predetermined antigen or closely related antigens (e.g., BSA, casein).

[0062] The term “inhibition” or “inhibition of…” refers to reducing a measurable amount or preventing it entirely.

[0063] The term "therapeuticly effective amount" refers to the amount of a conjugate that is effective in treating a disease or symptom in mammals. In the case of cancer, a therapeutically effective amount of a conjugate can reduce the number of cancer cells; reduce tumor size; inhibit (i.e., to some extent slow down and preferably stop) the invasion of cancer cells into surrounding organs; inhibit (i.e., to some extent slow down and preferably stop) tumor metastasis; inhibit tumor growth to some extent; and / or alleviate one or more symptoms associated with cancer to some extent. In terms of the extent to which the drug can inhibit growth and / or kill existing cancer cells, it can be cytoseptic and / or cytotoxic. For cancer treatment, efficacy can be measured, for example, by assessing time to progression (TTP) and / or determining the response rate (RR).

[0064] The term "basic" or "essentially" refers to the majority of a mixture or sample population, i.e., >50%, preferably more than 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% of the population.

[0065] The term "cytotoxic activity" refers to the cytotoxic effect of a drug, a camptothecin conjugate, or a camptothecin conjugate's intracellular metabolite. Cytotoxic activity can be expressed as the IC50 value, which is the concentration (moles or mass) of a drug that allows half of the cells to survive.

[0066] The term "cell inhibitory activity" refers to the antiproliferative effect of a drug, a camptothecin conjugate, or a camptothecin conjugate's intracellular metabolite.

[0067] As used herein, the term "cytotoxic agent" refers to a substance that has cytotoxic activity and causes cell destruction. This term is intended to include chemotherapeutic agents and toxins such as small molecule toxins or enzyme-active toxins of bacterial, fungal, plant, or animal origin, including their synthetic analogues and derivatives.

[0068] As used herein, the term "cell inhibitor" refers to a substance that inhibits cellular function, including substances that inhibit cell growth or proliferation. Cell inhibitors include inhibitors such as protein inhibitors, and for example, enzyme inhibitors. Cell inhibitors possess cellular inhibitory activity.

[0069] The terms “cancer” and “carcinoma” refer to or describe a physiological condition or disease in mammals that is typically characterized by uncontrolled cell growth. A “tumor” contains one or more cancer cells.

[0070] As used in this article, "autoimmune disease" refers to a disease or condition that originates from or targets an individual's own tissues or proteins.

[0071] As used herein, "patient" refers to a subject who has been given the camptothecin conjugate of the present invention. Patients include, but are not limited to, humans, rats, mice, guinea pigs, non-human primates, pigs, goats, cattle, horses, dogs, cats, birds, and poultry. Typically, patients are rats, mice, dogs, humans, or non-human primates, but more typically humans.

[0072] Unless the context otherwise indicates, the term "treat" or "treatment" refers to therapeutic and preventative treatment aimed at suppressing or slowing (alleviating) undesirable physiological changes or conditions, such as the development or spread of cancer. For the purposes of this invention, beneficial or desired clinical outcomes include, but are not limited to, reduction of symptoms, lessening of disease severity, stabilization of the disease state (i.e., non-deterioration), delay or slowing of disease progression, improvement or mitigation of the disease state, and remission (whether partial or complete), whether detectable or undetectable. "Treatment" may also imply a prolonged survival compared to expected survival without treatment. Those requiring treatment include both those already suffering from the disease or condition and those susceptible to it.

[0073] Within the context of cancer, the term "treatment" includes any or all of the following: killing tumor cells; inhibiting the growth of tumor cells, cancer cells, or tumors; inhibiting the replication of tumor cells or cancer cells; reducing the overall tumor burden or the number of cancer cells; and improving one or more symptoms of accompanying diseases.

[0074] Within the scope of autoimmune diseases, the term "treatment" includes any or all of the following: suppressing the replication of cells associated with an autoimmune disease state, including but not limited to cells that produce autoantibodies; reducing the autoantibody load; and improving one or more symptoms of the autoimmune disease.

[0075] As used herein, the term "compound" refers to and encompasses the compound itself, whether explicitly stated otherwise, unless the context clearly indicates that such salt forms are excluded. The term "compound" also encompasses the solvate form of the compound, wherein the solvent is non-covalently bonded to the compound or reversibly covalently bonded to the compound, such as when the carbonyl group of the compound is hydrated to form a geminal glycol. Solvate forms include the compound itself and its salt forms, and include hemisolvates, monosolvates, disolvates, including hydrates; when the compound can bind to two or more solvent molecules, the two or more solvent molecules can be the same or different.

[0076] In some cases, compounds of the present invention will include one or more of the expressly mentioned forms above, such as salts and solvates, which does not imply any solid form of the compound. However, such mentions are for emphasis only and should not be construed as excluding any other forms identified above. Furthermore, when the salt and / or solvate form of a compound or ligand pharmaceutical conjugate composition is not expressly mentioned, the omission should not be construed as excluding the salt and / or solvate form of the compound or conjugate unless the context clearly indicates the exclusion of such salt and / or solvate form.

[0077] As used herein, the phrase "its salt" refers to the salt form of a compound (e.g., a drug, a drug linker compound, or a ligand-drug conjugate compound). The salt form of a compound is one or more internal salt forms and / or involves the inclusion of another molecule such as an acetate ion, a succinate ion, or other counter ion. The counter ion of the salt form of a compound is typically an organic or inorganic moiety that stabilizes the charge on the parent compound. The salt form of a compound has one or more charged atoms in its structure. In cases where multiple charged atoms are part of the salt form, multiple counter ions and / or multiple charged counter ions are present. Therefore, the salt form of a compound typically has one or more charged atoms corresponding to those of the non-salt form of the compound and one or more counter ions. In some aspects, the non-salt form of a compound contains at least one amino or other basic moiety and thus, in the presence of an acid, yields an acid addition salt with a basic moiety. In other aspects, the non-salt form of a compound contains at least one carboxylic acid group or other acidic moiety and thus, in the presence of a base, yields a carboxylate or other anionic moiety. Exemplary salts include, but are not limited to, sulfates, trifluoroacetates, citrates, acetates, oxalates, chlorides, bromides, iodides, nitrates, bisulfates, phosphates, acid phosphates, isonicotinic acid esters, lactates, salicylates, citrates, tartrates, oleates, tannates, pantothenates, hydrogen tartrates, ascorbic acid salts, succinates, maleates, gentianates, fumarates, gluconates, glucurons, sucrose salts, formates, benzoates, glutamates, methanesulfonates, ethanesulfonates, benzenesulfonates, p-toluenesulfonates, and bis(hydroxynaphthyl)ates (i.e., 1,1'-methylenebis-(2-hydroxy-3-naphthylcarbamate)).

[0078] Pharmaceutically acceptable salts are salt forms of compounds suitable for administration to the subjects described herein, and in some respects include countercations or counteranions as described in PHStahl and CGWermuth, Handbook of Pharmaceutical Salts: Properties, Selection and Use, Weinheim / Zürich: Wiley-VCH / VHCA, 2002.

[0079] The linker unit is a bifunctional portion in a camptothecin conjugate that links camptothecin to a ligand unit. The linker unit of the present invention has several components (e.g., in some embodiments, an extended subunit having a basic unit; a connector unit that may or may not be present; a parallel connector unit that may or may not be present; a releasable linker; and a spacer subunit that may or may not be present).

[0080] As used herein, “PEG,” “PEG unit,” or “polyethylene glycol” refers to an organic moiety consisting of repeating ethylene-oxy subunits and can be polydisperse, monodisperse, or discrete (i.e., having a discrete number of ethylene-oxy subunits). Polydisperse PEG is a non-homogeneous mixture of sizes and molecular weights, while monodisperse PEG is typically purified from a non-homogeneous mixture and therefore has a single chain length and molecular weight. Preferred PEG units are discrete PEGs, which are compounds synthesized stepwise rather than via a polymerization process. Discrete PEGs provide a single molecule with a defined and specified chain length.

[0081] The PEG units provided herein comprise one or more polyethylene glycol chains, each consisting of one or more ethyleneoxy subunits covalently linked to each other. The polyethylene glycol chains may be linked together, for example, in a linear, branched, or star configuration. Typically, prior to introduction into the camptothecin conjugate, at least one polyethylene glycol chain is derivatized at one end with an electrophilically substituted alkyl portion to covalently link it to the urethane nitrogen of a methylene carbamate unit (i.e., representing an example of R). Typically, the terminal ethyleneoxy subunits in each polyethylene glycol chain that are not covalently linked to the remainder of the linking subunit are modified with a PEG end-capping unit, typically an optionally substituted alkyl group such as –CH3, CH2CH3, or CH2CH2CO2H. Preferred PEG units have a single polyethylene glycol chain having 4 to 24 –CH2CH2O- subunits covalently linked in series and terminated at one end by a PEG end-capping unit.

[0082] Unless otherwise specified, the term "alkyl" on its own or as part of another term refers to a substituted or unsubstituted straight-chain or branched, saturated or unsaturated hydrocarbon having a specified number of carbon atoms (e.g., "-C1-C8 alkyl" or "-C1-C...). 10"alkyl refers to an alkyl group having 1 to 8 or 1 to 10 carbon atoms. When the number of carbon atoms is not specified, the alkyl group has 1 to 8 carbon atoms. Representative straight-chain '-C1-C8 alkyl' groups include, but are not limited to, -methyl, -ethyl, -n-propyl, -n-butyl, -n-pentyl, -n-hexyl, -n-heptyl, and -n-octyl; while branched -C3-C8 alkyl groups include, but are not limited to, -isopropyl, -sec-butyl, -isobutyl, -tert-butyl, -isopentyl, and -2-methylbutyl; unsaturated -C2-C8 alkyl groups include, but are not limited to, -vinyl, - Allyl, -1-butenyl, -2-butenyl, -isobutenyl, -1-pentenyl, -2-pentenyl, -3-methyl-1-butenyl, -2-methyl-2-butenyl, -2,3-dimethyl-2-butenyl, -1-hexyl, 2-hexyl, -3-hexyl, -ethynyl, -propynyl, -1-butynyl, -2-butynyl, -1-pentynyl, -2-pentynyl, and -3-methyl-1-butynyl. Sometimes the alkyl group is unsubstituted. The alkyl group may be substituted by one or more groups. In other respects, the alkyl group will be saturated.

[0083] Unless otherwise specified, "alkylene" itself, or as part of another term, refers to a substituted or unsubstituted saturated, branched, straight-chain, or cyclic hydrocarbon group having the number of carbon atoms mentioned, typically 1-10, and having two monovalent groups at the center obtained by removing two hydrogen atoms from the same or two different carbon atoms of the parent alkane. Typical alkylene groups include, but are not limited to: methylene (CH2), 1,2-ethylene (CH2CH2), 1,3-propylene (CH2CH2CH2), 1,4-butylene (CH2CH2CH2CH2), etc. In a preferred aspect, the alkylene is a branched or straight-chain hydrocarbon (i.e., it is not a cyclic hydrocarbon).

[0084] Unless otherwise specified, "aryl" itself, or as part of another term, refers to a monovalent carbon cyclic aromatic hydrocarbon group having the referred number of carbon atoms, typically 6-20, obtained by removing a hydrogen atom from a single carbon atom of the parent aromatic ring system, whether substituted or unsubstituted. Some aryl groups are represented by "Ar" in exemplary structures. Typical aryl groups include, but are not limited to, groups derived from benzene, substituted benzene, naphthalene, anthracene, biphenyl, etc. An exemplary aryl group is a phenyl group.

[0085] Unless otherwise stated, "arylene" itself or as part of another term refers to an aryl group as defined above that has two covalent bonds (i.e., it is divalent) and can be oriented in the ortho, meta, or para positions as shown in the structure below, with phenyl as an example group:

[0086]

[0087] Unless otherwise specified, "C3-C8 heterocycle" itself, or as part of another term, refers to a monocyclic or non-aromatic monocyclic or bicyclic ring system having 3 to 8 carbon atoms (also called ring members) and one to four independent heteroatom ring members selected from N, O, P, or S, obtained by removing a hydrogen atom from a ring atom of the parent ring system, whether substituted or unsubstituted. One or more N, C, or S atoms in the heterocycle may be oxidized. The ring containing the heteroatom can be aromatic or non-aromatic. A heterocycle in which all ring atoms are involved in the structure of an aromatic compound is called a heteroaryl ring; otherwise, it is called a heterocarbon ring.

[0088] Unless otherwise specified, heterocycles are attached to their side groups at any heteroatom or carbon atom that will produce a stable structure. Thus, heteroaryl groups can be bonded to the aromatic carbons of their aromatic ring system, referred to as C-linked heteroaryl groups, or to non-double-bonded nitrogen atoms (i.e., not =N-) in their aromatic ring system, referred to as N-linked heteroaryl groups. Therefore, nitrogen-containing heterocycles can be C-linked or N-linked and include pyrrole moieties such as pyrrole-1-yl (N-linked) and pyrrole-3-yl (C-linked), and imidazole moieties such as imidazole-1-yl and imidazole-3-yl (both N-linked) and imidazole-2-yl, imidazole-4-yl, and imidazole-5-yl moieties (all C-linked).

[0089] Unless otherwise specified, “C3-C8 heteroaryl” refers to an aromatic C3-C8 heterocycle, where the subscript indicates the total number of carbons in the cyclic system of the heterocycle or the total number of aromatic carbons in the aromatic ring system of the heteroaryl group, and does not imply the size of the ring system or the presence or absence of ring fusion. Representative examples of C3-C8 heterocycles include, but are not limited to, pyrrolidinyl, azacyclic butyl, piperidinyl, morpholinyl, tetrahydrofuranyl, tetrahydropyranyl, benzofuranyl, benzothiophene, indolyl, benzopyrazolyl, pyrrolidinyl, thiophene, furanyl, thiazolyl, imidazolyl, pyrazolyl, pyrimidinyl, pyridinyl, pyrazinyl, isothiazolyl, and isoxazolyl.

[0090] When explicitly given, the size of a heterocyclic or heteroaryl ring system is described by the total number of atoms in the ring. For example, specifying a 5- or 6-membered heteroaryl indicates the total number of aromatic atoms (i.e., 5 or 6) in the heteroaromatic ring system of the heteroaryl, but does not imply the number of aromatic heteroatoms or aromatic carbons in the ring system. Fused heteroaryls are explicitly referred to or so implied by the context, and are generally described by the number of aromatic atoms in each aromatic ring fused together to form a fused heteroaromatic ring system. For example, a 5,6-membered heteroaryl is an aromatic 5-membered ring fused to an aromatic 6-membered ring, where one or both rings have one or more aromatic heteroatoms or where heteroatoms are shared between the two rings.

[0091] A heterocycle that is fused with an aryl or heteroaryl group through connection to a non-aromatic portion of a fused ring system, thereby maintaining the heterocycle's non-aromatic nature and being part of a larger structure, is an example of an optionally substituted heterocycle, wherein the heterocycle is replaced by ring fusion with the aryl or heteroaryl group. Similarly, an aryl or heteroaryl group fused with a heterocycle or carbocyclic ring that is part of a larger structure through connection to an aromatic portion of a fused ring system is an example of an optionally substituted aryl or heterocycle, wherein the aryl or heterocycle is replaced by ring fusion with the heterocycle or carbocyclic ring.

[0092] Unless otherwise stated, "C3-C8 heterocyclo" itself, or as part of another term, refers to the C3-C8 heterocyclic group as defined above, in which one of the hydrogen atoms of the heterocycle is replaced by a bond (i.e., it is divalent). Unless otherwise stated, "C3-C8 heteroaryl" itself, or as part of another term, refers to the C3-C8 heteroaryl group as defined above, in which one of the hydrogen atoms of the heteroaryl group is replaced by a bond (i.e., it is divalent).

[0093] Unless otherwise specified, "C3-C8 carbocyclic ring" itself, or as part of another term, refers to a 3-, 4-, 5-, 6-, 7-, or 8-membered, monovalent, substituted or unsubstituted, saturated or unsaturated non-aromatic monocyclic or bicyclic carbocyclic ring obtained by removing a hydrogen atom from a ring atom of the parent ring system. Representative C3-C8 carbocyclic rings include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclopentadienyl, cyclohexyl, cyclohexenyl, 1,3-cyclohexadienyl, 1,4-cyclohexadienyl, cycloheptyl, 1,3-cycloheptadienyl, 1,3,5-cycloheptanetrienyl, cyclooctyl, and cyclooctadienyl.

[0094] Unless otherwise indicated, “C3-C8 carbocyclic group” by itself or as part of another term refers to another C3-C8 carbocyclic group as defined above, in which the hydrogen atom of the carbocyclic group is replaced by a bond (i.e., it is divalent).

[0095] Unless otherwise specified, the term "heteroalkyl" itself, or in combination with another term, unless otherwise stated, refers to a stable straight-chain or branched hydrocarbon or combination thereof that is fully saturated or contains 1 to 3 degrees of unsaturation, consisting of the number of carbon atoms mentioned and one to ten, preferably one to three, heteroatoms selected from O, N, Si, and S, wherein nitrogen and sulfur atoms may optionally be oxidized and nitrogen heteroatoms may optionally be quaternized. One or more heteroatoms O, N, and S may be located at any internal position of the heteroalkyl group or at the position where the alkyl group is attached to the rest of the molecule. Heteroatoms Si may be located at any position of the heteroalkyl group, including the position where the alkyl group is attached to the rest of the molecule.

[0096] Examples include –CH2-CH2-O-CH3, -CH2-CH2-NH-CH3, -CH2-CH2-N(CH3)-CH3, -CH2-S-CH2-CH3, -CH2-CH2-S(O)-CH3, -NH-CH2-CH2-NH-C(O)-CH2-CH3, -CH2-CH2-S(O)2-CH3, -CH=CH-O-CH3, -Si(CH3)3, -CH2-CH=NO-CH3, and –CH=CH-N(CH3)-CH3. At most two heteroatoms can be consecutive, such as, for example, -CH2-NH-OCH3 and –CH2-O-Si(CH3)3. Typically, C1 to C4 heteroalkyl or heteroalkylene groups have 1 to 4 carbon atoms and 1 or 2 heteroatoms, while C1 to C3 heteroalkyl or heteroalkylene groups have 1 to 3 carbon atoms and 1 or 2 heteroatoms. In some respects, heteroalkyl and heteroalkylene compounds are saturated.

[0097] Unless otherwise specified, the term "heteroalkyl" on its own, or in combination with another term, refers to a divalent group derived from a heteroalkyl group (as discussed above), such as –CH2-CH2-S-CH2-CH2- and –CH2-S-CH2-CH2-NH-CH2-. For heteroalkyl groups, the heteroatom may also occupy any one or both of the chain ends. Furthermore, the orientation of the linking group is not implied for alkylene and heteroalkylene linkages.

[0098] Unless otherwise specified, "aminoalkyl" on its own or in combination with another term refers to a heteroalkyl group in which the alkyl moiety as defined herein is replaced by an amino, alkylamino, dialkylamino, or cycloalkylamino group. Exemplary non-limiting aminoalkyl groups include –CH2NH2, -CH2CH2NH2, -CH2CH2NHCH3, and -CH2CH2N(CH3)2, and also include branched types such as –CH(CH3)NH2 and -C(CH3)CH2NH2 in (R)- or (S)- configurations. Alternatively, aminoalkyl can be an alkyl moiety, group, or substituent as defined herein, wherein the sp(s) of the radical carbon are... 3 The carbon has been replaced by an amino or alkylamino moiety, in which its sp 3 nitrogen replaced alkyl sp 3 Carbon, provided that at least one sp is retained. 3 Carbon. When the aminoalkyl moiety is referred to as a substituent of a larger structure or another moiety, the aminoalkyl moiety is covalently attached to that structure or moiety via a carbon radical of the alkyl moiety of the aminoalkyl moiety.

[0099] Unless otherwise stated, “alkylamino” and “cycloalkylamino” on their own or in combination with another term refer to an alkyl or cycloalkyl group as described herein, wherein the radical carbon of the alkyl or cycloalkyl group has been replaced by a nitrogen radical, provided that at least one sp is retained. 3 Carbon. In those cases where the alkylamino group is replaced at its nitrogen atom by another alkyl moiety, the resulting substituted group is sometimes referred to as a dialkylamino moiety, group, or substituent, wherein the alkyl moiety replacing the nitrogen atom is chosen independently.

[0100] Exemplary and non-limiting amino, alkylamino, and dialkylamino substituents include those having a –N(R')2 structure, wherein R' in these examples is independently selected from hydrogen or C. 1-6 Alkyl groups are typically hydrogen or methyl, while in cycloalkylamines containing heterocyclic alkyl groups, the two R's and the nitrogen atoms to which they are attached define the heterocyclic ring. When both R's are hydrogen or alkyl, the moiety is sometimes described as a primary amine group and a tertiary amine group, respectively. When one R' is hydrogen and the other is alkyl, the moiety is sometimes described as a secondary amine group. Primary and secondary alkylamine moieties are more reactive as nucleophiles to electrophilic centers containing carbonyl groups, while tertiary amines are more basic.

[0101] "Substituted alkyl" and "substituted aryl" refer to alkyl and aryl groups, respectively, in which one or more hydrogen atoms (usually one) are independently replaced by substituents. Typical substituents include, but are not limited to, -X, -R', -OH, -OR', -SR', -N(R')2, -N(R')3, =NR', -CX3, -CN, -NO2, -NR'C(=O)R', -C(=O)R', -C(=O)N(R')2, -S(=O)2R', -S(=O)2NR', -S(=O)R', -OP(=O)(OR')2, -P(=O)(OR')2, -PO3 ＝ , PO3H2, -C(=O)R', -C(=S)R', -CO2R', -CO2 - -C(=S)OR', -C(=O)SR', -C(=S)SR', -C(=O)N(R')2, -C(=S)N(R')2 and -C(=NR)N(R')2, wherein each X is independently selected from halogens: -F, -Cl, -Br and -I; and each R' is independently selected from -H, -Cl-C 20 Alkyl, -C6-C 20 Aryl, -C3-C 14 Heterocyclic rings, protecting groups, and prodrug moieties.

[0102] More typically, the substituents are selected from -X, -R', -OH, -OR', -SR', -N(R')2, -N(R')3, =NR', -NR'C(=O)R', -C(=O)R', -C(=O)N(R')2, -S(=O)2R', -S(=O)2NR', -S(=O)R', -C(=O)R', -C(=S)R', -C(=O)N(R')2, -C(=S)N(R')2, and -C(=NR)N(R')2, wherein each X is independently selected from –F and -Cl, or selected from -X, -R', -OH, -OR', -N(R')2, -N(R')3, -NR'C(=O)R', -C(=O)N(R')2, -S(=O)2R', -S(=O)2NR', -S(=O)R', -C(=O)R', -C(=O)N(R')2, -C(=NR)N(R')2, protecting groups, and prodrug moieties, wherein each X is –F; and wherein each R' is independently selected from hydrogen, -Cl-C 20 Alkyl, -C6-C 20 Aryl, -C3-C 14 Heterocyclic rings, protecting groups, and prodrug moieties.

[0103] In some respects, the alkyl substituent is selected from -N(R')2, -N(R')3 and -C(=NR)N(R')2, wherein R' is selected from hydrogen and -C1-C 20 Alkyl groups. In other respects, the alkyl group is defined by a series of ethyleneoxy groups. Alkylenes, carbocyclics, carbocyclic groups, arylenes, heteroalkyl groups, heteroalkylenes, heterocyclic groups, heteroaryl groups, and heteroarylenes, as described above, may also be similarly substituted.

[0104] As used in this article, a "protecting group" refers to the part that prevents or reduces the ability of the atoms or functional groups attached to it to participate in undesirable reactions. Greene (1999), "P ROTECTIVE G ROUPS I N O RGANIC S YNTHESIS The third edition of Wiley Interscience provides typical protecting groups for atoms or functional groups. In some cases, protecting groups are used against heteroatoms such as oxygen, sulfur, and nitrogen to minimize or avoid undesirable reactions with electrophilic compounds. In other cases, protecting groups are used to reduce or eliminate the nucleophilicity and / or basicity of unprotected heteroatoms. Non-limiting examples of protected oxygen are given by -OR. PR Given, where R PRThe protecting group is the hydroxyl group, which is typically protected in the form of an ester (e.g., acetate, propionate, or benzoate). Other protecting groups on the hydroxyl group will prevent interference with the nucleophilicity of organometallic reagents or other highly basic reagents, where the hydroxyl group is typically protected in the form of an ether, including alkyl or heterocyclic alkyl ethers (e.g., methyl or tetrahydropyranyl ethers), alkoxymethyl ethers (e.g., methoxymethyl or ethoxymethyl ethers), optionally substituted aryl ethers, and silyl ethers (e.g., trimethylsilyl (TMS), triethylsilyl (TES), tert-butyldiphenylsilyl (TBDPS), tert-butyldimethylsilyl (TBS / TBDMS), triisopropylsilyl (TIPS), and [2-(trimethylsilyl)ethoxy]-methylsilyl (SEM)). Nitrogen protecting groups include those for primary or secondary amines, such as those in -NHR. PR or -N(R) PR In )2-, where R PR At least one of them is a nitrogen-protecting group or two R groups. PR Together they form a protecting group.

[0105] A protecting group is suitable when it can prevent or avoid undesirable side reactions or premature loss of the protecting group during the purification of the newly formed molecule under the reaction conditions required to achieve the desired chemical transformation elsewhere in the molecule, and when desired, and can be removed without adversely affecting the structural or stereochemical integrity of the newly formed molecule. By way of example and not limitation, suitable protecting groups may include those described above for protecting functional groups. Suitable protecting groups are sometimes those used in peptide coupling reactions.

[0106] "Aromatic alcohol" itself, or as part of a larger structure, refers to an aromatic ring system substituted with a hydroxyl functional group (-OH). Therefore, an aromatic alcohol is any aryl, heteroaryl, arylene, or heteroarylene moiety as described herein, having a hydroxyl functional group bonded to an aromatic carbon in its aromatic ring system. When the aromatic ring system of an aromatic alcohol is a substituent of this moiety, the aromatic alcohol can be part of a larger portion, or can be inserted into a larger portion via ring fusion, and can optionally be substituted with a moiety as described herein, including one or more other hydroxyl substituents. Phenolic alcohols are aromatic alcohols having a phenolic group as an aromatic ring.

[0107] "Aliphatic alcohol" refers, either on its own or as part of a larger structure, to a portion of a non-aromatic carbon bonded to a hydroxyl functional group (-OH). The hydroxyl-bearing carbon may be unsubstituted (i.e., methanol), or may have one, two, or three optionally substituted branched or unbranched alkyl substituents defining a primary alcohol, or an aliphatic secondary or tertiary alcohol, within a linear or cyclic structure. When part of a larger structure, the alcohol may be a substituent of the structure substituted via a hydroxyl-bearing carbon, via an alkyl or other moiety as described herein, or via a substituent of that alkyl or other moiety. Aliphatic alcohols encompass non-aromatic cyclic structures (i.e., carbocyclic and heterocyclic rings, optionally substituted) in which the hydroxyl functional group is bonded to a non-aromatic carbon of its cyclic ring system.

[0108] As used herein, “arylalkyl” or “heteroarylalkyl” refers to a substituent, part, or group wherein the aryl moiety is bonded to the alkyl moiety, i.e., aryl-alkyl-, wherein the alkyl and aryl groups are as described above, for example, C6H5-CH2- or C6H5-CH(CH3)CH2-. Arylalkyl or heteroarylalkyl groups are defined by the sp... 3 Carbon is combined with larger structures or parts.

[0109] As used herein, an "electron-withdrawing group" refers to a functional group or electronegative atom that inductively and / or resonantly withdraws electrons from the atoms to which it is bonded, either inductively or resonantly, which can be more dominant (i.e., the functional group or atom may inductively withdraw electrons, but overall may donate electrons via resonance) and tends to stabilize the anion or electron-rich moiety. The electron-withdrawing effect is typically transmitted inductively (albeit in a decaying form) to other atoms connected to the bonded atom, which is now electron-deficient due to the electron-withdrawing group (EWG), thus affecting the electrophilicity of more distant reaction centers. Exemplary electron-withdrawing groups include, but are not limited to, -C(=O), -CN, -NO2, -CX3, -X, -C(=O)OR', -C(=O)N(R')2, -C(=O)R', -C(=O)X, -S(=O)2R', -S(=O)2OR', -S(=O)2NHR', -S(=O)2N(R')2, -P(=O)(OR')2, -P(=O)(CH3)NHR', -NO, and -N(R')3. + Where X is -F, -Br, -Cl, or -I, and in some respects, R' is independently selected from hydrogen and C each time it appears. 1-6 Alkyl groups, and certain O-linked moieties such as acyloxy groups as described herein.

[0110] Exemplary EWGs may also include aryl groups (e.g., phenyl), depending on the substitutions and certain heteroaryl groups (e.g., pyridine). Therefore, the term "electron-withdrawing group" also includes aryl or heteroaryl groups further substituted with electron-withdrawing groups. Typically, the electron-withdrawing group on the aryl or heteroaryl group is -C (=O), -CN, -NO2, -CX3, and –X, where X is independently selected from halogens, typically -F or -Cl. Depending on its substituents, the alkyl moiety may also be an electron-withdrawing group.

[0111] "Leaving group capability" refers to the ability of an alcoholic, thiol-containing, amine-containing, or amide-containing compound corresponding to camptothecin in a camptothecin conjugate to be released as a free drug from the conjugate upon activation of a self-immolative event within the conjugate. This release may be variable without the benefit of a methylene carbamate unit to which its camptothecin is attached (i.e., when camptothecin is directly attached to the self-immolative portion and does not have an inserted methylene carbamate unit). A good leaving group is typically a weak base, and the more acidic the functional group expended from such a conjugate, the weaker the base of the conjugate. Therefore, in the absence of a methylene carbamate unit (i.e., where camptothecin is directly attached to the self-immolative portion), the leaving group capability of the free drug containing an alcoholic, thiol-containing, amine-containing, or amide from the camptothecin will be related to the pKa of the drug functional group expended from the conjugate. Thus, a lower pKa of the functional group will increase its leaving group capability. While other factors may contribute to the release of the free drug from the conjugate without the benefits of methylene carbamates, generally, drugs with functional groups having lower pKa values ​​are better leaving groups than those linked via functional groups having higher pKa values. Another consideration is that functional groups with excessively low pKa values ​​may lead to unacceptable activity properties due to the premature loss of camptothecin via spontaneous hydrolysis. For conjugates employing methylene carbamate units, a universal functional group (i.e., carbamic acid) with a pKa value that allows for efficient release of the free drug is generated upon self-digestion, without suffering unacceptable camptothecin loss.

[0112] As used herein, the term "succinimide moiety" refers to an organic moiety consisting of a succinimide ring system present in an extended subunit (Z) of a type, which typically also includes an alkylene portion of the imide nitrogen bonded to the ring system. The succinimide moiety is typically generated by Michael addition of the thiol group of the ligand unit to the maleimide ring system of the extended subunit precursor (Z'). Thus, the succinimide moiety is composed of a sulfur-substituted succinimide ring system, and when present in a camptothecin conjugate, its imide nitrogen is substituted by the remainder of the linking subunit of the camptothecin conjugate and optionally by one or more substituents present on the maleimide ring system of Z'.

[0113] As used herein, “acid-amide moiety” refers to succinic acid with an amide substituent, which is generated by the hydrolysis of a sulfo-substituted succinimide ring system of the succinimide moiety, resulting in the breaking of one of its carbonyl-nitrogen bonds. The linker unit provided by the hydrolysis of the succinic acid-amide moiety is unlikely to suffer premature loss of the ligand unit bonded to it via the elimination of the antibody-sulfo substituent. The hydrolysis of the succinimide ring system of the sulfo-substituted succinimide moiety is expected to provide regiochemical isomers of the acid-amide moiety, as the difference in reactivity between the two carbonyl carbons of the succinimide ring system can be attributed at least in part to any substituents present in the maleimide ring system of the extended subunit precursor and the sulfo substituent introduced by the targeting ligand.

[0114] As used herein, the term "prodrug" refers to a compound with low or no biological activity that is converted in vivo into a more biologically active compound via a chemical or biological process (i.e., a chemical reaction or enzymatic biotransformation). Typically, the biological activity of a biologically active compound is reduced (i.e., converted into a prodrug) by chemically modifying the compound with a prodrug portion. In some respects, prodrugs are type II prodrugs, which are bioactivated extracellularly (e.g., in digestive fluids) or in the human circulatory system (e.g., in the blood). Exemplary prodrugs are esters and β-D-glucopyranoside.

[0115] In many cases, the assemblies of conjugates, linkers, and components described herein will refer to reactive groups. A “reactive group” or RG is a group containing a reactive site (RS) capable of forming a bond with a component of the linker unit (i.e., A, W, Y) or camptothecin D. RS is the reactive site within the reactive group (RG). Reactive groups include thiol groups forming disulfide or thioether bonds, aldehyde, ketone, or hydrazine groups forming hydrazone bonds, carboxyl or amino groups forming peptide bonds, carboxyl or hydroxyl groups forming ester bonds, sulfonic acids forming sulfonamide bonds, alcohols forming carbamate bonds, and amines forming sulfonamide or carbamate bonds.

[0116] The table below is a schematic of reactive groups, reactive sites, and exemplary functional groups that may be formed after the reaction at the reactive sites. This table is not limiting. Those skilled in the art will understand that the R' and R” portions mentioned in the table are in fact any organic portion (e.g., alkyl groups, aryl groups, heteroaryl groups, or substituted alkyl, aryl, or heteroaryl groups) that is compatible with the bond formation provided in converting RG into one of the exemplary functional groups. It should also be understood that, as applicable to embodiments of the invention, R' may represent one or more components of a self-stabilizing linker or optional auxiliary linker, and R” may represent one or more components of an optional auxiliary linker, camptothecin, stabilizing unit, or detection unit.

[0117] Detailed Implementation Plan

[0118] Many embodiments of the invention are described below, which are not intended to limit the invention in any way, and are followed by a more detailed discussion of the components constituting the conjugates. Those skilled in the art will understand that each conjugate identified and any selected embodiment thereof is intended to encompass the full scope of each component and linker.

[0119] Camptothecin conjugate

[0120] In one main embodiment, a camptothecin conjugate or a salt thereof having the following formula is provided:

[0121] L-(QD) p

[0122] in

[0123] L stands for ligand unit;

[0124] The subscript p is an integer from 1 to 16;

[0125] Q is a connection subunit having a formula selected from the following:

[0126] -ZA-,-ZA-RL-,-ZA-RL-Y-,ZAS * -W-,

[0127] ZAS * -RL-,-ZAB(S * )-RL-,-ZAS * -W-RL-,-ZAS * -RL-Y-,

[0128] and -ZAB(S * )-RL-Y-,

[0129] Where Z is the stretcher unit.

[0130] A stands for key or connector unit;

[0131] B stands for Parallel connector unit;

[0132] S * It is a partitioning agent;

[0133] W stands for amino acid unit;

[0134] RL is a releasable unit;

[0135] Y represents the spacer unit; and

[0136] D is a drug unit selected from the following:

[0137]

[0138]

[0139] in

[0140] R B The derivative is selected from H, C1-C8 alkyl, C1-C8 haloalkyl, C3-C8 cycloalkyl, (C3-C8 cycloalkyl)-C1-C4 alkyl, phenyl, and phenyl-C 1- The C4 alkyl portion;

[0141] R C Selected from C1-C6 alkyl and C 3- The C6 cycloalkyl moiety;

[0142] R F and R F’ Each is independently selected from -H, C1-C8 alkyl, C1-C8 hydroxyalkyl, C1-C8 aminoalkyl, (C1-C4 alkylamino)-C1-C8 alkyl, N,N-(C1-C4 hydroxyalkyl)(C1-C4 alkyl)amino-C1-C8 alkyl, N,N-bis(C1-C4 alkyl)amino-C 1- C8 alkyl, N-C1-C4 hydroxyalkyl-C1-C8 aminoalkyl, C1-C8 alkyl C(O)-, C1-C8 hydroxyalkyl-C(O)-, C1-C8 aminoalkyl-C(O)-, C3-C 10 cycloalkyl, (C3-C 10 Cycloalkyl)-C1-C4 alkyl, C3-C 10 Heterocyclic alkyl groups, (C3-C 10 The portions of heterocyclic alkyl-C1-C4 alkyl, phenyl, phenyl-C1-C4 alkyl, diphenyl-C1-C4 alkyl, heteroaryl, and heteroaryl-C1-C4 alkyl; or

[0143] R F and R F’ It combines with the nitrogen atoms to form a 5-, 6-, or 7-membered ring having 0 to 3 substituents selected from halogens, C1-C4 alkyl groups, -OH, -OC1-C4 alkyl groups, -NH2, and -NHC. 1- C4 alkyl and -N(C1-C4 alkyl)2; and wherein R B R C R F and R F’The cycloalkyl, heterocycloalkyl, phenyl, and heteroaryl moieties are composed of 0 to 3 elements selected from halogens, C 1- C4 alkyl, -OH, -OC1-C4 alkyl, -NH2, -NHC 1- C4 alkyl and -N(C 1- Substituents of C4 alkyl)2; and

[0144] Where Q is -ZA-RL-, -ZA-RL-Y-, -ZAS * -RL-、-ZAB(S * -RL-、-ZAS * -RL-Y- or -ZAB(S) * When RL is any of the releasable linkers disclosed herein, the connection point between D and Q is via a heteroatom of any of the hydroxyl, primary, or secondary amine functional groups present on CPT1, CPT2, CPT3, CPT4, CPT5, CPT6, or CPT7, or

[0145] Where Q is -ZA- or -ZAS * -W- or -ZAB(S * -W-, or when Q is -ZAS * -RL-、-ZAB(S * -RL-、-ZAS * -W-RL- or -ZAB(S * When RL is a releasable unit other than a glucuronide unit, the connection point between D and Q is through the oxygen atom of the hydroxyl substituent in the lactone ring of CPT1, CPT2, CPT3, CPT4, CPT5, CPT6, or CPT7; and

[0146] The condition is that when the linkage point is the nitrogen atom of a CPT6 amino group, R F and R F’ At least one of them is -H,

[0147] The condition is that when D is a CPT1 with a amino group linked to it, -ZA-RL-, -ZA-RL-Y-, -ZAS * -RL-、-ZAB(S * -RL-、-ZAS * -RL-Y- and -ZAB(S * The -ZA- of )-RL-Y- is different from the succinimide-hexanoyl-β-alanyl group which optionally has a succinimide ring in a hydrolyzed form.

[0148] In one set of implementations, D has the formula CPT5.

[0149] In one set of implementations, D has the formula CPT2.

[0150] In one set of implementations, D has the formula CPT3.

[0151] In one set of implementations, D has the formula CPT4.

[0152] In one set of implementations, D has the formula CPT1.

[0153] In one set of implementations, D has the formula CPT6.

[0154] In one set of implementations, D has the formula CPT7.

[0155] In one set of implementations, Q has a form selected from the following:

[0156] -ZA-RL- and -ZA-RL-Y-,

[0157] Wherein RL is a releasable linker as a glucuronide unit, and groups Z, A, and Y have the meanings provided above and in any of the embodiments specifically described herein.

[0158] In one set of implementations, Q has a form selected from the following:

[0159] -ZAS * -RL- and -ZAS * -RL-Y-,

[0160] RL is a releasable linker that serves as a glucuronide unit, and the groups Z, A, and S... * Y has the meanings provided above and in any of the embodiments specifically described herein.

[0161] In one set of implementations, Q has a form selected from the following:

[0162] -ZAB(S * -RL- and -ZAB(S * )-RL-Y-,

[0163] RL is a releasable linker that serves as a glucuronide unit, and the groups Z, A, and S... * B and Y have the meanings provided above and in any of the embodiments specifically described herein.

[0164] In another set of implementations, Q has a form selected from the following:

[0165] -ZA- or -ZA-RL-,

[0166] Wherein RL is a releasable linker other than the glucuronide unit, and groups Z and A have the meanings provided above and in any of the embodiments specifically described herein.

[0167] In another set of implementations, Q has a form selected from the following:

[0168] -ZAS * -RL- and -ZAB(S * )-RL-

[0169] Where RL is a releasable linker other than the glucuronide unit, and the groups Z, A, and S * B has the meanings provided above and in any of the embodiments specifically described herein.

[0170] In another set of implementations, Q has a form selected from the following:

[0171] -ZAS * -W- and -ZAB(S * )-W-,

[0172] Among them, the groups Z, A, and S * B and W have the meanings provided above and in any of the embodiments specifically described herein.

[0173] In another set of implementations, Q has a form selected from the following:

[0174] -ZAS * -W-RL- and -ZAB(S * )-W-RL-,

[0175] Where RL is a releasable linker other than the glucuronide unit, and the groups Z, A, and S * B and W have the meanings provided above and in any of the embodiments specifically described herein.

[0176] In one set of implementations, Q has the formula -ZA-RL-, -ZA-RL-Y-, -ZAS * -RL-, -ZAS*-RL-Y-, -ZAB(S) * -RL- or –ZAB(S * Camptothecin conjugates consisting of drug units having the formula CPT1 are represented by the following formula:

[0177]

[0178]

[0179] Specifically, RL is any of the releasable linkers disclosed herein, preferably RL is a glucuronide unit, and the groups L, Z, A, S * B and Y have the meanings provided above and in any of the embodiments specifically described herein, provided that the -ZA- of formulas CPT1iN, CPT1iiN, CPT1iiiN, CPT1ivN, CPT1vN and CPT1viN is different from the succinimide-hexanoyl-β-alanyl group optionally having a succinimide ring in a hydrolyzed form.

[0180] In other embodiments, camptothecin conjugates comprising the formula -ZA-, -ZA-RL-, -ZAS*-W-, -ZAB(S*)-W-, -ZAS*-RL-, –ZAB(S*)-RL-, -ZAS*-W-RL-, and –ZAB(S*)-W-RL- and composed of pharmaceutical units having the formula CPT1 are represented by the following formula:

[0181]

[0182]

[0183] Specifically, RL is a releasable linker other than the glucuronide unit, and the groups L, Z, A, and S... * B and W have the meanings provided above and in any of the embodiments specifically described herein.

[0184] In another set of embodiments, the camptothecin conjugate, wherein Q has the formula -ZA-RL-, -ZA-RL-Y-, -ZAS*-RL-, -ZAS*-RL-Y-, -ZAB(S*)-RL-, or –ZAB(S*)-RL-Y- and is composed of pharmaceutical units having the formula CPT2, is represented by the following formula:

[0185]

[0186] Specifically, RL is any of the releasable linkers disclosed herein, preferably RL is a glucuronide unit, and the groups L, Z, A, S * B and Y have the meanings provided above and in any of the embodiments specifically described herein.

[0187] In other embodiments, camptothecin conjugates comprising pharmaceutical units having the formula -ZA-, -ZA-RL-, -ZAS*-W-, -ZAB(S*)-W-, -ZAS*-RL-, –ZAB(S*)-RL-, -ZAS*-W-RL-, and –ZAB(S*)-W-RL- and consisting of pharmaceutical units having the formula CPT2 are represented by the following formula:

[0188]

[0189] Specifically, RL is a releasable linker other than the glucuronide unit, and the groups L, Z, A, and S... * B and W have the meanings provided above and in any of the embodiments specifically described herein.

[0190] In one set of implementations, R in formulas CPT2iOa, CPT2iiOa, CPT2iiiOa, CPT2ivOa, CPT2vOa, CPT2viOa, CPT2iOb, CPT2iiOb, CPT2iiiOb, CPT2ivOb, CPT2vOb, CPT2viOb, CPT2viiOb, or CPT2viiiOb B It is a portion selected from -H, C1-C8 alkyl and C1-C8 haloalkyl.

[0191] In another set of embodiments, R in formulas CPT2iOa, CPT2iiOa, CPT2iiiOa, CPT2ivOa, CPT2vOa, CPT2viOa, CPT2iOb, CPT2iiOb, CPT2iiiOb, CPT2ivOb, CPT2vOb, CPT2viOb, CPT2viiOb, or CPT2viiiOb B It is a portion selected from C3-C8 cycloalkyl, (C3-C8 cycloalkyl)-C1-C4 alkyl-, phenyl, and phenyl-C1-C4 alkyl-, and wherein R B The cycloalkyl and phenyl moieties are substituted with 0-3 substituents selected from halogen, C1-C4 alkyl, -OH, -OC1-C4 alkyl, -NH2, -NHC1-C4 alkyl and -N(C1-C4 alkyl)2.

[0192] In another set of embodiments, the camptothecin conjugate, wherein Q has the formula -ZA-RL-, -ZA-RL-Y-, -ZAS*-RL-, -ZAS*-RL-Y-, -ZAB(S*)-RL-, or –ZAB(S*)-RL-Y- and is composed of pharmaceutical units having the formula CPT3, is represented by the following formula:

[0193]

[0194]

[0195] Specifically, RL is any of the releasable linkers disclosed herein, preferably RL is a glucuronide unit, and the groups L, Z, A, S * B and Y have the meanings provided above and in any of the embodiments specifically described herein.

[0196] In other embodiments, camptothecin conjugates comprising the formula -ZA-, -ZA-RL-, -ZAS*-W-, -ZAB(S*)-W-, -ZAS*-RL-, –ZAB(S*)-RL-, -ZAS*-W-RL-, and –ZAB(S*)-W-RL- and composed of pharmaceutical units having the formula CPT3 are represented by the following formula:

[0197]

[0198] Specifically, RL is a releasable linker other than the glucuronide unit, and the groups L, Z, A, and S... * B and W have the meanings provided above and in any of the embodiments specifically described herein.

[0199] In one set of implementations, R in formulas CPT3iOa, CPT3iiOa, CPT3iiiOa, CPT3ivOa, CPT3vOa, CPT3viOa, CPT3iO'a, CPT3iiO'a, CPT3iiiO'a, CPT3ivO'a, CPT3vO'a, CPT3viO'a, CPT3iOb, CPT3iiOb, CPT3iiiOb, CPT3ivOb, CPT3vOb, CPT3viOb, CPT3viiOb, or CPT3viiiOb C It is a C1-C6 alkyl group.

[0200] In one set of implementations, R in formulas CPT3iOa, CPT3iiOa, CPT3iiiOa, CPT3ivOa, CPT3vOa, CPT3viOa, CPT3iO'a, CPT3iiO'a, CPT3iiiO'a, CPT3ivO'a, CPT3vO'a, CPT3viO'a, CPT3iOb, CPT3iiOb, CPT3iiiOb, CPT3ivOb, CPT3vOb, CPT3viOb, CPT3viiOb, or CPT3viiiOb C It is a C3-C6 cycloalkyl group.

[0201] In another set of embodiments, the camptothecin conjugate, wherein Q has the formula -ZA-RL-, -ZA-RL-Y-, -ZAS*-RL-, -ZAS*-RL-Y-, -ZAB(S*)-RL-, or –ZAB(S*)-RL-Y- and is composed of pharmaceutical units having the formula CPT4, is represented by the following formula:

[0202]

[0203]

[0204] Specifically, RL is any of the releasable linkers disclosed herein, preferably RL is a glucuronide unit, and the groups L, Z, A, S * B and Y have the meanings provided above and in any of the embodiments specifically described herein.

[0205] In other embodiments, camptothecin conjugates comprising the formula -ZA-, -ZA-RL-, -ZAS*-W-, -ZAB(S*)-W-, -ZAS*-RL-, –ZAB(S*)-RL-, -ZAS*-W-RL-, and –ZAB(S*)-W-RL- and composed of pharmaceutical units having the formula CPT4 are represented by the following formula:

[0206]

[0207]

[0208] Specifically, RL is a releasable linker other than the glucuronide unit, and the groups L, Z, A, and S... * B and W have the meanings provided above and in any of the embodiments specifically described herein.

[0209] In another set of embodiments, the camptothecin conjugate, wherein Q has the formula -ZA-RL-, -ZA-RL-Y-, -ZAS*-RL-, -ZAS*-RL-Y-, -ZAB(S*)-RL-, or –ZAB(S*)-RL-Y- and is composed of pharmaceutical units having the formula CPT5, is represented by the following formula:

[0210]

[0211]

[0212] Specifically, RL is any of the releasable linkers disclosed herein, preferably RL is a glucuronide unit, and the groups L, Z, A, S *B and Y have the meanings provided above and in any of the embodiments specifically described herein.

[0213] In other embodiments, camptothecin conjugates comprising the formula -ZA-, -ZA-RL-, -ZAS*-W-, -ZAB(S*)-W-, -ZAS*-RL-, –ZAB(S*)-RL-, -ZAS*-W-RL-, and –ZAB(S*)-W-RL- and composed of pharmaceutical units having the formula CPT5 are represented by the following formula:

[0214]

[0215]

[0216] Specifically, RL is a releasable linker other than the glucuronide unit, and the groups L, Z, A, and S... * B and W have the meanings provided above and in any of the embodiments specifically described herein.

[0217] In another set of embodiments, the camptothecin conjugate, wherein Q has the formula -ZA-RL-, -ZA-RL-Y-, -ZAS*-RL-, -ZAS*-RL-Y-, -ZAB(S*)-RL-, or –ZAB(S*)-RL-Y- and is composed of pharmaceutical units having the formula CPT6, is represented by the following formula:

[0218]

[0219]

[0220] Specifically, RL is any of the releasable linkers disclosed herein, preferably RL is a glucuronide unit, and the groups L, Z, A, S * B and Y have the meanings provided above and in any of the embodiments specifically described herein.

[0221] In other embodiments, camptothecin conjugates comprising pharmaceutical units having the formula -ZA-, -ZA-RL-, -ZAS*-W-, -ZAB(S*)-W-, -ZAS*-RL-, –ZAB(S*)-RL-, -ZAS*-W-RL-, and –ZAB(S*)-W-RL- are represented by the following formula:

[0222]

[0223]

[0224] Specifically, RL is a releasable linker other than the glucuronide unit, and the groups L, Z, A, and S... * B and W have the meanings provided above and in any of the embodiments specifically described herein.

[0225] In one set of implementations, R in formula CPT6iN, CPT6iiN, CPT6iiiN, CPT6ivN, CPT6vN or CPT6viN F Yes, it's -H.

[0226] In one set of implementations, R in formulas CPT6iOa, CPT6iiOa, CPT6iiiOa, CPT6ivOa, CPT6vOa, CPT6viOa, CPT6iOb, CPT6iiOb, CPT6iiiOb, CPT6ivOb, CPT6vOb, CPT6viOb, CPT6viiOb, or CPT6viiiOb F and R F’ Both are -H.

[0227] In one set of implementations, R in formula CPT6iN, CPT6iiN, CPT6iiiN, CPT6ivN, CPT6vN or CPT6viN F It is a part selected from the following group: C1-C8 alkyl, C1-C8 hydroxyalkyl, C1-C8 aminoalkyl, (C1-C4 alkylamino)-C1-C8 alkyl-, N,N-(C1-C4 hydroxyalkyl)(C1-C4 alkyl)amino-C 1- C8 alkyl-, N,N-bis(C1-C4 alkyl)amino-C1-C8 alkyl-, N-(C1-C4 hydroxyalkyl)-C1-C8 aminoalkyl-, C1-C8 alkyl-C(O)-, C1-C8 hydroxyalkyl-C(O)-, and C1-C8 aminoalkyl-C(O)-.

[0228] In one set of implementations, R in formula CPT6iN, CPT6iiN, CPT6iiiN, CPT6ivN, CPT6vN or CPT6viN F This is a selection from the following group: C3-C 10 cycloalkyl, (C3-C 10 Cycloalkyl)-C1-C4 alkyl-, C3-C 10 Heterocyclic alkyl groups, (C3-C 10 Heterocyclic alkyl)-C1-C4 alkyl-, phenyl, phenyl-C1-C4 alkyl-, diphenyl-C1-C4 alkyl-, heteroaryl and heteroaryl-C1-C4 alkyl-, and wherein R FThe cycloalkyl, heterocycloalkyl, phenyl, and heteroaryl moieties are substituted by 0-3 substituents independently selected from the group consisting of halogen, C1-C4 alkyl, -OH, -OC1-C4alkyl, -NH2, -NHC1-C4alkyl, and -N(C1-C4alkyl)2.

[0229] In one set of implementations, R in formula CPT6iN, CPT6iiN, CPT6iiiN, CPT6ivN, CPT6vN or CPT6viN F It is selected from the following group: -H, C3-C 10 cycloalkyl, (C3-C 10 Cycloalkyl)-C1-C4 alkyl-, C3-C 10 Heterocyclic alkyl groups, (C3-C 10 Heterocyclic alkyl)-C1-C4 alkyl, phenyl, phenyl-C1-C4 alkyl-, diphenyl-C1-C4 alkyl, heteroaryl and heteroaryl-C1-C4 alkyl-, and wherein R F The cycloalkyl, heterocycloalkyl, phenyl, and heteroaryl moieties are substituted by 0-3 substituents independently selected from the group consisting of halogen, C1-C4 alkyl, -OH, -OC1-C4 alkyl, -NH2, -NHC1-C4 alkyl, and -N(C1-C4 alkyl)2.

[0230] In one set of implementations, R in formulas CPT6iOa, CPT6iiOa, CPT6iiiOa, CPT6ivOa, CPT6vOa, CPT6viOa, CPT6iOb, CPT6iiOb, CPT6iiiOb, CPT6ivOb, CPT6vOb, CPT6viOb, CPT6viiOb, or CPT6viiiOb F and R F’ It combines with the nitrogen atoms to form a 5-, 6-, or 7-membered ring having 0 to 3 substituents selected from: halogens, C1-C4 alkyl groups, -OH, -OC1-C4 alkyl groups, -NH2, and -NHC. 1- C4 alkyl and -N(C 1- C4 alkyl)2.

[0231] In one set of implementations, R in formulas CPT6iOa, CPT6iiOa, CPT6iiiOa, CPT6ivOa, CPT6vOa, CPT6viOa, CPT6iOb, CPT6iiOb, CPT6iiiOb, CPT6ivOb, CPT6vOb, CPT6viOb, CPT6viiOb, or CPT6viiiOb F and R F’At least one of them is a moiety independently selected from the group consisting of: C1-C8 alkyl, C1-C8 hydroxyalkyl, C1-C8 aminoalkyl, (C1-C4 alkylamino)-C 1- C8 alkyl, N,N-(C 1- C4 hydroxyalkyl)(C 1- C4 alkyl)amino-C 1- C8 alkyl-,N,N-di(C 1- C4 alkyl)amino-C 1- C8 alkyl-, N-(C 1- (C4-hydroxyalkyl)-C1-C8-aminoalkyl-, C1-C8-alkylC(O)-, C1-C8-hydroxyalkyl-C(O)-, and C1-C8-aminoalkyl-C(O)-, and the others are portions selected from the group consisting of: –H, C1-C8-alkyl, C1-C8-hydroxyalkyl, C1-C8-aminoalkyl, (C1-C4-alkylamino)-C1-C8-alkyl-, N,N-(C1-C4-hydroxyalkyl)(C1-C4-alkyl)amino-C 1- C8 alkyl-, N,N-bis(C1-C4 alkyl)amino-C1-C8 alkyl-, N-(C1-C4 hydroxyalkyl)-C1-C8 aminoalkyl-, C1-C8 alkyl-C(O)-, C1-C8 hydroxyalkyl-C(O)-, and C1-C8 aminoalkyl-C(O)-.

[0232] In one set of implementations, each R in formula CPT6iO, CPT6iiO, CPT6iiiO, CPT6ivO, CPT6vO or CPT6viO F and R F’ It is an independent selection from the following group: C 1- C8 alkyl, C 1- C8 hydroxyalkyl, C 1- C8 aminoalkyl, (C 1- C4 alkylamino)-C 1- C8 alkyl-, N,N-(C1-C4 hydroxyalkyl)(C 1- C4 alkyl)amino-C1-C8 alkyl-, N,N-bis(C 1- C4 alkyl)amino-C1-C8 alkyl-, N-(C1-C4 hydroxyalkyl)-C 1- C8 aminoalkyl, C 1- C8 alkyl-C(O)-, C1-C8 hydroxyalkyl-C(O)-, and C1-C8 aminoalkyl-C(O)-.

[0233] In one set of implementations, R in formula CPT6iO, CPT6iiO, CPT6iiiO, CPT6ivO, CPT6vO or CPT6viO F and R F’At least one of them is a part independently selected from the following groups: C3-C 10 cycloalkyl, C3-C 10 Cycloalkyl-C1-C4 alkyl-, C3-C 10 Heterocyclic alkyl groups, (C3-C 10 Heterocyclic alkyl)-C1-C4 alkyl-, phenyl, phenyl-C1-C4 alkyl, diphenyl-C1-C4 alkyl, heteroaryl and heteroaryl-C1-C4 alkyl-, and wherein R F Or R F’ The cycloalkyl, heterocycloalkyl, phenyl, and heteroaryl moieties are substituted by 0-3 substituents independently selected from the group consisting of: halogen, C1-C4 alkyl, -OH, -OC1-C4 alkyl, -NH2, -NHC 1- C4 alkyl and -N(C 1- C4 alkyl)2, and the others are selected from the group consisting of: –H, C1-C8 alkyl, C1-C8 hydroxyalkyl, C1-C8 aminoalkyl, (C1-C4 alkylamino)-C1-C8 alkyl-, N,N-(C1-C4 hydroxyalkyl)(C1-C4 alkyl)amino-C 1- C8 alkyl-, N,N-bis(C1-C4 alkyl)amino-C1-C8 alkyl-, N-(C1-C4 hydroxyalkyl)-C1-C8 aminoalkyl-, C1-C8 alkyl-C(O)-, C1-C8 hydroxyalkyl-C(O)-, and C1-C8 aminoalkyl-C(O)-)2.

[0234] In one set of implementations, R in formula CPT6iO, CPT6iiO, CPT6iiiO, CPT6ivO, CPT6vO or CPT6viO F and R F’ At least one of them is a part independently selected from the following groups: C3-C 10 cycloalkyl, C3-C 10 Cycloalkyl-C1-C4 alkyl-, C3-C 10 Heterocyclic alkyl groups, (C3-C 10 Heterocyclic alkyl)-C1-C4 alkyl-, phenyl, phenyl-C 1- C4 alkyl, diphenyl C 1- C4 alkyl, heteroaryl and heteroaryl-C 1- C4 alkyl-, and the others are selected from the group consisting of: –H, C3-C 10 cycloalkyl, (C3-C 10 Cycloalkyl)-C1-C4 alkyl-, C3-C 10 Heterocyclic alkyl groups, (C3-C 10 Heterocyclic alkyl)-C1-C4 alkyl-, phenyl, phenyl-C1-C4 alkyl-, diphenyl-C1-C4 alkyl-, heteroaryl and heteroaryl-C1-C4 alkyl-, wherein RF Or R F’ The cycloalkyl, heterocycloalkyl, phenyl, and heteroaryl moieties are substituted by 0-3 independent substituents selected from the group consisting of: halogen, C 1- C4 alkyl, -OH, -OC 1- C4 alkyl, -NH2, -NHC 1- C4 alkyl and -N(C1-C4 alkyl)2.

[0235] In one set of implementations, each R in formula CPT6iO, CPT6iiO, CPT6iiiO, CPT6ivO, CPT6vO, or CPT6viO F and R F’ It is a part that is independently selected from the following group: -H, C3-C 10 cycloalkyl, (C3-C 10 Cycloalkyl)-C1-C4 alkyl-, C3-C 10 Heterocyclic alkyl groups, (C3-C 10 Heterocyclic alkyl)-C1-C4 alkyl-, phenyl, phenyl-C1-C4 alkyl-, diphenyl-C1-C4 alkyl-, heteroaryl and heteroaryl-C1-C4 alkyl-, and wherein R F Or R F’ The cycloalkyl, heterocycloalkyl, phenyl, and heteroaryl moieties are substituted by 0-3 independent substituents selected from the group consisting of: halogen, C 1- C4 alkyl, -OH, -OC 1- C4 alkyl, -NH2, -NHC1-C alkyl and -N(C1-C4 alkyl)2.

[0236] In another set of embodiments, the camptothecin conjugate, wherein Q has the formula -ZA-RL-, -ZA-RL-Y-, -ZAS*-RL-, -ZAS*-RL-Y-, -ZAB(S*)-RL-, or –ZAB(S*)-RL-Y- and is composed of pharmaceutical units having the formula CPT7, is represented by the following formula:

[0237]

[0238]

[0239]

[0240] Specifically, RL is any of the releasable linkers disclosed herein, preferably RL is a glucuronide unit, and the groups L, Z, A, S * B and Y have the meanings provided above and in any of the embodiments specifically described herein.

[0241] In other embodiments, camptothecin conjugates comprising the formula -ZA-, -ZA-RL-, -ZAS*-W-, -ZAB(S*)-W-, -ZAS*-RL-, –ZAB(S*)-RL-, -ZAS*-W-RL-, and –ZAB(S*)-W-RL- and composed of pharmaceutical units having the formula CPT5 are represented by the following formula:

[0242]

[0243]

[0244] Specifically, RL is a releasable linker other than the glucuronide unit, and the groups L, Z, A, and S... * B and W have the meanings provided above and in any of the embodiments specifically described herein.

[0245] Camptothecin-linker compounds

[0246] In some embodiments, when preparing camptothecin conjugates, a complete drug-linker assembly needs to be synthesized before conjugation with a target agent. In such embodiments, the camptothecin-linker compound as described herein is an intermediate compound. In those embodiments, the elongated subunit in the camptothecin-linker compound is not yet covalently linked to the ligand unit (i.e., is the elongated subunit precursor Z') and therefore has functional groups for conjugation with the target ligand. In one embodiment, the camptothecin-linker compound consists of camptothecin (shown herein as formulas CPT1, CPT2, CPT3, CPT4, CPT5, CPT6, and CPT7) and a linker unit (Q) containing a glucuronide unit as a releasable linker (RL) through which the ligand unit is linked to camptothecin.

[0247] In another embodiment, the camptothecin-linker compound comprises a camptothecin compound of formula CPT1, CPT2, CPT3, CPT4, CPT5, CPT6, or CPT7, and a linker unit (Q) comprising a releasable linker (RL) different from a glucuronide unit, through which the ligand unit is conjugated to the camptothecin compound. Thus, in another embodiment, in addition to RL, the linker unit also comprises an extension subunit precursor (Z') containing functional groups for conjugation with a target agent (which is a precursor to the ligand unit) and thus enabling the RL to be linked to the ligand unit (directly or indirectly). In some of those embodiments, a parallel linker unit (B) may be present when it is desired to add a dispensing agent (S*) as a side-chain attachment. In any of those embodiments, a linker unit (A) is present when it is desired to increase the distance between the extension subunit and RL.

[0248] In one set of embodiments, the camptothecin-linker compound comprises a camptothecin compound having the formula CPT1, CPT2, CPT3, CPT4, CPT5, CPT6 or CPT7 and a linker unit (Q), wherein Q comprises a releasable linker (RL) as a glucuronide unit directly linked to an extended subunit precursor (Z') or indirectly linked to Z' through a linker unit of the camptothecin-linker compound via one or more intermediate components (i.e., A, S* and / or B(S*)) of the linker unit, wherein Z' is composed of functional groups capable of forming covalent bonds with the target agent.

[0249] In another set of embodiments, the camptothecin-linker compound consists of camptothecin having the formula CPT1, CPT2, CPT3, CPT4, CPT5, CPT6 or CPT7 and a linker unit (Q), wherein Q comprises a releasable linker (RL) other than a glucuronide unit, which is directly linked to the extended subunit precursor (Z') or indirectly linked to Z' through a linker unit of the camptothecin-linker compound via one or more intermediate components (i.e., A, S* and / or B(S*)) of the linker unit, wherein Z' is composed of functional groups capable of forming covalent bonds with the target agent.

[0250] In the context of camptothecin conjugates and / or camptothecin-linker compounds, assemblies are preferably described according to their constituent functional groups. Although some procedures are also described herein, those skilled in the art will well understand the order of assembly and the general conditions for the preparation of conjugates and compounds.

[0251] Component groups

[0252] Ligand unit:

[0253] In some embodiments of the invention, a ligand unit is present. The ligand unit (L-) is a targeting agent that specifically binds to a target moiety. In one set of embodiments, the ligand unit specifically and selectively binds to a cellular component (cell-binding agent) or another target molecule of interest. The role of the ligand unit is to target and present camptothecin (CPT1, CPT2, CPT3, CPT4, CPT5, CPT6, or CPT7) to a specific target cell population that interacts with the ligand unit due to the presence of its target component or molecule, allowing the free drug to be subsequently released within (i.e., intracellularly) or nearby (i.e., extracellularly) the target cells. The ligand unit L includes, but is not limited to, proteins, polypeptides, and peptides. Suitable ligand units include, for example, antibodies, such as full-length antibodies and their antigen-binding fragments, interferons, lymphokines, hormones, growth factors and colony-stimulating factors, vitamins, nutrient transport molecules (such as, but not limited to, transferrin), or any other cell-binding molecule or substance. In some embodiments, the ligand unit (L) is derived from an antibody or non-antibody protein targeting agent.

[0254] In one set of embodiments, the ligand unit is bonded to a Q (linker unit) comprising a glucuronide-releasing linker. As described above, other linking components may also be present in the conjugates described herein to provide additional spacing between the camptothecin drug compound and the ligand unit (e.g., an extension unit and optionally a linker unit A) or to provide compositional properties to increase solubility (e.g., a dispensing agent S*). In some of these embodiments, the ligand unit is bonded to the Z of the linker unit via a heteroatom of the ligand unit. Heteratomes that may be present on the ligand unit for this bonding include sulfur (in one embodiment, from a thiol group of the targeting ligand), oxygen (in one embodiment, from a carboxyl or hydroxyl group of the targeting ligand), and nitrogen, optionally substituted (in one embodiment, from a primary or secondary amine functional group of the targeting ligand, or in another embodiment, from an optionally substituted amide nitrogen). These heteroatoms may be present on the targeting ligand in their native state (e.g., in naturally occurring antibodies) or may be introduced into the targeting ligand via chemical modification or bioengineering.

[0255] In one embodiment, the target agent, which serves as a precursor to the ligand unit, has a thiol functional group, such that the ligand unit is bonded to the linker unit via the sulfur atom of the thiol functional group.

[0256] In another embodiment, the target agent, which is a precursor of the ligand unit, has one or more lysine residues that are capable of reacting with an activated ester (such esters include, but are not limited to, N-hydroxysuccinimide, pentafluorophenyl, and p-nitrophenyl esters) of the extended subunit precursor of the camptothecin-linker compound intermediate and thus providing an amide bond consisting of the nitrogen atom of the ligand unit and the C=O group of the extended subunit of the linker unit.

[0257] In another aspect, the target agent serving as a precursor to the ligand unit has one or more lysine residues that can be chemically modified to introduce one or more thiol groups. In those embodiments, the ligand unit is covalently linked to the linker unit via the sulfur atom of the thiol functional group. Reagents that can be used to modify lysine in this manner include, but are not limited to, N-succinimide-S-acetylthioacetate (SATA) and 2-iminothiacyclopentane hydrochloride (Traut reagent).

[0258] In another embodiment, the target agent serving as a precursor to the ligand unit has one or more carbohydrate groups that can be modified to provide one or more thiol functional groups. In the camptothecin conjugate, the chemically modified ligand unit is bonded to the linker subunit component (e.g., the extended subunit) via the sulfur atom of the thiol functional group.

[0259] In yet another embodiment, the target agent serving as a precursor to the ligand unit has one or more carbohydrate groups that can be oxidized to provide an aldehyde (-CHO) functional group (see, for example, Laguzza et al., 1989, J. Med. Chem. 32(3): 548-55). In these embodiments, the corresponding aldehyde interacts with a reactive site on the elongation unit precursor to form a bond between the elongation unit and the ligand unit. Reactive sites on the elongation unit precursor capable of interacting with reactive carbonyl-containing functional groups on the target ligand unit include, but are not limited to, hydrazine and hydroxylamine. Other schemes for modifying proteins to link linker units (Q) or related substances are described in Coligan et al., Current Protocols in Protein Science, vol. 2, John Wiley & Sons (2002) (incorporated herein by reference).

[0260] In some aspects, the target agent, as a precursor of the ligand unit, can form a covalent bond between the extended subunit (Z) and the ligand unit whose structure corresponds to the target agent by interacting with reactive functional groups on the extended subunit precursor (Z'). The functional groups of Z' capable of interacting with the target agent will depend on the nature of the target agent whose structure corresponds to the ligand unit. In some embodiments, the reactive group is maleimide, which is present on the extended subunit before being linked to form the ligand unit (i.e., the maleimide portion of the extended subunit precursor). The covalent connection between the ligand unit and the extended subunit is achieved by the interaction of the thiol functional group of the target agent, as a precursor of the ligand unit, with the maleimide functional group of Z' to form a sulfur-substituted succinimide. The thiol functional group may be present on the target agent in its native state (e.g., in naturally occurring residues) or may be introduced into the target agent via chemical modification or bioengineering.

[0261] In yet another embodiment, the ligand unit is derived from the antibody, and the thiol group is generated by the reduction of the antibody's interchain disulfide. Accordingly, in some embodiments, the linker unit is conjugated to a cysteine ​​residue from one or more reduced interchain disulfides.

[0262] In yet another embodiment, the ligand unit is derived from the antibody, and the thiol functional group is chemically introduced into the antibody, for example, through the introduction of a cysteine ​​residue. Accordingly, in some embodiments, the linker unit (with or without a linker, camptothecin) is conjugated to the ligand unit via the cysteine ​​residue introduced by the ligand unit.

[0263] For bioconjugates, the site of drug conjugation has been observed to affect numerous parameters, including ease of conjugation, drug-linker stability, impact on the biophysical properties of the resulting bioconjugate, and in vitro cytotoxicity. Regarding drug-linker stability, in some cases, the conjugation site between the drug-linker moiety and the ligand unit can affect the ability of the conjugated drug-linker moiety to undergo elimination reactions, leading to premature release of the free drug. Conjugation sites on the target ligand include, for example, reduced interchain disulfides and selected cysteine ​​residues at engineered sites. In some embodiments, the conjugation method for forming camptothecin conjugates as described herein uses thiol residues at genetically engineered sites (e.g., position 239 according to the EU index as described in Kabat) that are less prone to elimination reactions compared to conjugation methods using thiol residues from reduced disulfide bonds. In other embodiments, the conjugation method for forming camptothecin conjugates as described herein uses thiol residues generated from the reduction of interchain disulfide bonds.

[0264] In some embodiments, the camptothecin conjugate comprises a non-immunoreactive protein, polypeptide, or peptide as its ligand unit. Accordingly, in some embodiments, the ligand unit is derived from a non-immunoreactive protein, polypeptide, or peptide. Examples include, but are not limited to, transferrin, epidermal growth factor (“EGF”), dermalin, gastrin, gastrin-releasing peptide, platelet-derived growth factor, IL-2, IL-6, transforming growth factor (“TGF”) such as TGF-α and TGF-β, vaccinia virus growth factor (“VGF”), insulin and insulin-like growth factor I and II, somatostatin, lectins, and apoproteins derived from low-density lipoprotein.

[0265] Particularly preferred ligand units are derived from antibodies. Therefore, in any embodiment described herein, the ligand unit is derived from an antibody. Available polyclonal antibodies are a heterogeneous population of antibody molecules derived from the serum of immunized animals. Available monoclonal antibodies are a homogeneous population of antibodies against specific antigenic determinants (e.g., cancer cell antigens, viral antigens, microbial antigens, proteins, peptides, carbohydrates, chemicals, nucleic acids, or fragments thereof). In some embodiments, monoclonal antibodies (mAbs) against antigens of interest are prepared using any techniques known in the art, which provide the production of antibody molecules through continuous cell lines in culture.

[0266] Available monoclonal antibodies include, but are not limited to, human monoclonal antibodies, humanized monoclonal antibodies, or chimeric human-mouse (or other species) monoclonal antibodies. Antibodies include full-length antibodies and their antigen-binding fragments. Human monoclonal antibodies can be prepared by any of a variety of techniques known in the art (e.g., Teng et al., 1983, Proc. Natl. Acad. Sci. USA. 80:7308-7312; Kozbor et al., 1983, Immunology Today 4:72-79; and Olsson et al., 1982, Meth. Enzymol. 92:3-16).

[0267] The antibodies that can be used to carry out the present invention are intact antibodies or functionally active fragments, derivatives, or analogs of antibodies, wherein the antibodies or fragments thereof are capable of immune-specific binding to target cells (e.g., cancer cell antigens, viral antigens, or microbial antigens) or other antibodies that bind to tumor cells or matrix. In this respect, “functionally active” means that the fragments, derivatives, or analogs are capable of immune-specific binding to target cells. In order to determine which CDR sequences will bind to the antigen, in some embodiments, synthetic peptides containing CDR sequences are used in the binding assay of the antigen by any binding assay method known in the art (e.g., BIA core assay) (see, for example, Kabat et al., 1991, Sequences of Proteins of Immunological Interest, 5th ed., National Institute of Health, Bethesda, Md.; Kabat E et al., 1980, J. Immunology 125(3):961-969).

[0268] Other available antibodies include fragments of antibodies, such as, but not limited to, F(ab')2 fragments, Fab fragments, Fvs, single-chain antibodies, bi-chain antibodies, triple-chain antibodies, quadruple-chain antibodies, scFv, scFv-FV, or any other molecule with the same specificity as the antibody.

[0269] Additionally, in some embodiments, recombinant antibodies containing human and non-human portions, such as chimeric and humanized monoclonal antibodies, prepared using standard recombinant DNA technology, are available. Chimeric antibodies are molecules in which the different portions are derived from different animal species, such as those having, for example, a variable region derived from a mouse monoclonal antibody and a constant region of a human immunoglobulin. (See, for example, U.S. Patent Nos. 4,816,567 and 4,816,397, which are incorporated herein by reference in their entirety). Humanized antibodies are antibody molecules derived from non-human species that have one or more complementarity-determining regions (CDRs) derived from the non-human species and a framework region derived from a human immunoglobulin molecule. (See, for example, U.S. Patent No. 5,585,089, which is incorporated herein by reference in its entirety). In some embodiments, such chimeric and humanized monoclonal antibodies are produced using recombinant DNA techniques known in the art, for example using International Publication No. WO 87 / 02671; European Patent Publication No. 0 184 187; European Patent Publication No. 0 171 496; European Patent Publication No. 0 173 494; International Publication No. WO 86 / 01533; US Patent No. 4,816,567; Berter et al., Science (1988) 240:1041-1043; Liu et al., Proc. Nat'l. Acad. Sci. USA (1987) 84:3439-3443; Liu et al., J. Immunol. (1987) 139:3521-3526; Sun et al., Proc. Nat'l. Acad. Sci. USA (1987) 84:214-218; Nishimura et al., Cancer. Res. (1987) 47:999-1005; Wood et al., Nature (1985) 314:446-449; Shaw et al., J. Nat'l. Cancer The methods described in Inst. (1988) 80:1553-1559; Morrison, Science (1985) 229:1202-1207; Oi et al., BioTechniques (1986) 4:214-221; US ​​Patent No. 5,225,539; Jones et al., Nature (1986) 321:552-525; Verhoeyan et al., Science (1988) 239:1534-1536; and Beidler et al., J. Immunol. (1988) 141:4053-4060 are all incorporated herein by reference in their entirety.

[0270] In some cases (e.g., when immunogenicity to non-human or chimeric antibodies may occur), fully human antibodies are preferred and are produced in some embodiments using transgenic mice that are unable to express endogenous immunoglobulin heavy and light chain genes but are able to express human heavy and light chain genes.

[0271] Antibodies include modified analogs and derivatives, i.e., analogs and derivatives modified by covalent linkage of any type of molecule, as long as such covalent linkage allows the antibody to maintain its antigen-binding immune specificity. For example, but not limited to, antibody derivatives and analogs include those that have been further modified, such as by glycosylation, acetylation, PEGylation, phosphorylation, amidation, derivatization by known protecting / blocking groups, proteolytic cleavage, linkage to cellular antibody units or other proteins, etc. In some embodiments, one or more of these numerous chemical modifications are performed using known techniques, including but not limited to specific chemical cleavage, acetylation, formylation, metabolic synthesis in the presence of tunicamycin, etc. In other embodiments, antibody analogs or derivatives contain one or more non-natural amino acids, which are sometimes used in conjunction with one or more of the aforementioned chemical modifications.

[0272] In some embodiments, the antibody may have one or more modifications (e.g., substitution, deletion, or addition) in the amino acid residues that interact with the Fc receptor. Those modifications include amino acid residues identified as being involved in the interaction between the anti-Fc domain and the FcRn receptor (see, for example, International Publication No. WO 97 / 34631, which is incorporated herein by reference in its entirety).

[0273] In some implementations, antibodies with immune specificity against cancer cell antigens are commercially available or produced using methods known to those skilled in the art, such as recombinant expression techniques. The nucleotide sequences encoding antibodies with immune specificity against cancer cell antigens are sometimes obtained, for example, from GenBank or similar databases, literature publications, or through routine cloning and sequencing.

[0274] In one specific implementation, known antibodies used to treat cancer may be used.

[0275] In another specific embodiment, the compositions and methods according to the invention use antibodies for treating autoimmune diseases.

[0276] In some embodiments, the available antibody binds to a receptor or receptor complex expressed on activated lymphocytes. In some embodiments, the receptor or receptor complex may comprise an immunoglobulin gene superfamily member, a TNF receptor superfamily member, an integrin, a cytokine receptor, a chemokine receptor, a major histocompatibility protein, a lectin, or a complement regulatory protein.

[0277] In some implementations, the antibody introduced into the camptothecin conjugate will specifically bind to CD19, CD30, CD33, CD70, or LIV-1.

[0278] Camptothecin compounds:

[0279] The camptothecin compounds used in the various implementation schemes described herein are represented by the following formula:

[0280]

[0281]

[0282] R B The derivative is selected from H, C1-C8 alkyl, C1-C8 haloalkyl, C3-C8 cycloalkyl, (C3-C8 cycloalkyl)-C1-C4 alkyl, phenyl, and phenyl-C 1- The C4 alkyl portion;

[0283] R C Selected from C1-C6 alkyl and C 3- The C6 cycloalkyl moiety;

[0284] R F and R F’ Each is independently selected from -H, C1-C8 alkyl, C1-C8 hydroxyalkyl, C1-C8 aminoalkyl, (C1-C4 alkylamino)-C1-C8 alkyl, N,N-(C1-C4 hydroxyalkyl)(C1-C4 alkyl)amino-C1-C8 alkyl, N,N-bis(C1-C4 alkyl)amino-C 1- C8 alkyl, N-C1-C4 hydroxyalkyl-C1-C8 aminoalkyl, C1-C8 alkyl C(O)-, C1-C8 hydroxyalkyl-C(O)-, C1-C8 aminoalkyl-C(O)-, C3-C 10 cycloalkyl, (C3-C 10 Cycloalkyl)-C1-C4 alkyl, C3-C 10 Heterocyclic alkyl groups, (C3-C 10 The portions of heterocyclic alkyl-C1-C4 alkyl, phenyl, phenyl-C1-C4 alkyl, diphenyl-C1-C4 alkyl, heteroaryl, and heteroaryl-C1-C4 alkyl; or

[0285] R F and R F’ It combines with the nitrogen atoms respectively to form a 5-, 6-, or 7-membered ring having 0 to 3 substituents selected from halogens, C1-C4 alkyl, -OH, -OC1-C4 alkyl, -NH2, -NHC1-C4 alkyl, and -N(C1-C4 alkyl)2;

[0286] Where R BR C R F and R F’ The cycloalkyl, heterocycloalkyl, phenyl, and heteroaryl moieties are composed of 0 to 3 elements selected from halogens, C 1- C4 alkyl, -OH, -OC1-C4 alkyl, -NH2, -NHC1-C4 alkyl, and -N(C 1- Substituents of C4 alkyl)2.

[0287] Other camptothecin compounds used in the context of the camptothecin conjugates and camptothecin linker compounds described herein are camptothecin compounds 14a-14z of Table I and compounds 18a-18r of Table J, as well as camptothecin compounds having a pentacyclic or hexacyclic fused skeleton with structures similar to those provided by formulas CPT1, CPT2, CPT3, CPT4, CPT5, CPT6, CPT7, 14a-14z, and 18a-18r, which in some embodiments have other groups, including but not limited to hydroxyl, thiol, amine, or amide functional groups, whose oxygen, sulfur, or optionally substituted nitrogen atoms are capable of binding to the linker and being released as a free drug from the camptothecin conjugate. In some embodiments, this functional group provides a unique site on the camptothecin compound that can be used to link to the linker unit (Q). The resulting drug-linker moiety of the camptothecin conjugate is a compound capable of releasing an active free drug at the site targeted by its ligand unit to exert cytotoxic, cell-inhibiting, or immunosuppressive effects.

[0288] "Free drug" refers to a drug because it exists once released from the drug-linker portion. In some embodiments, the free drug comprises a fragment of a releasable linker or spacer unit (Y) group. The free drug comprising the releasable linker or spacer unit (Y) fragment is released from the remaining drug-linker portion via cleavage of the releasable linker, or via cleavage of the bonds in the spacer unit (Y) group, and is biologically active after release. In some embodiments, the free drug differs from a conjugated drug in that the functional group of the free drug used for attachment to the self-dissolving assembly unit is no longer conjugated with the components of the camptothecin conjugate (except for previously shared heteroatoms). For example, the free hydroxyl functional group of an alcohol-containing drug may be represented by DO*H, while in the conjugated form, an oxygen heteroatom represented by O* is introduced into the methylene carbamate unit of the self-dissolving unit. After the activation of the self-dissolving portion and the release of the free drug, the covalent bond with O* will be replaced by a hydrogen atom, so that the oxygen heteroatom represented by O* exists on the free drug in the form of -OH.

[0289] Connecting subunit (Q)

[0290] As described above, in some embodiments, the connecting subunit (Q) has a formula selected from the following:

[0291] -ZA-RL-;-ZA-RL-Y-;-ZAS * -RL-;-ZAB(S * )-RL-;

[0292] -ZAS * -RL-Y-; and -ZAB(S * )-RL-Y-;

[0293] Where Z represents an extension sub-unit; A represents a key or connector unit; B represents a branch unit; S * RL is a dispensing agent; RL is a releasable linker as a glucuronide; and Y is a spacer unit; and

[0294] The connection point between D and Q is achieved through a hydroxyl group and any heteroatom of a primary or secondary amine present on any of CPT1, CPT2, CPT3, CPT4, CPT5, CPT6 or CPT7, or compounds 14a-14z in Table I and compounds 18a-18r in Table J.

[0295] In other embodiments, the connection subunit (Q) has a formula selected from the following:

[0296] -ZA-;-ZA-RL-;-ZAS * -W-;-ZAB(S * )-W-;-ZAS * -RL-;-ZAB(S * )-RL-;

[0297] -ZAS * -W-RL-; and -ZAB(S * )-W-RL-;

[0298] Where Z represents an extended sub-unit; A represents a key or connector unit; B represents a parallel connector unit; S * For dispensing agents; RL is a releasable linker other than the glucuronic acid unit; and W is an amino acid unit; and

[0299] The connection point of Q is achieved through a hydroxyl substituent on the lactone ring of any of CPT1, CPT2, CPT3, CPT4, CPT5, CPT6 or CPT7, or compounds 14a-14z in Table I and compounds 18a-18r in Table J.

[0300] In one set of implementations, Q has a form selected from the following: -ZAS * -RL- and -ZAS * -RL-Y-.

[0301] In another set of embodiments, Q has a form selected from the following: -ZAB(S * -RL- and -ZAB(S * )-RL-Y-.

[0302] In yet another set of implementations, Q has a formula selected from the following: -ZA-RL- and -ZA-RL-Y-.

[0303] Extended subunit (Z) or (Z'):

[0304] The extension subunit (Z) is a component of the camptothecin conjugate or camptothecin-linker compound or other intermediate, and its function is to link the ligand unit to the rest of the conjugate. In this respect, the extension subunit has functional groups that can form bonds with functional groups of the target ligand before being linked to the ligand unit (i.e., the extension subunit precursor Z').

[0305] In some embodiments, the elongation unit precursor (Z') has an electrophilic group capable of interacting with a reactive nucleophilic group present on the ligand unit (e.g., antibody) to provide a covalent bond between the ligand unit and the elongation unit of the linker unit. Nucleophilic groups on the antibody with this capability include, but are not limited to, thiol, hydroxyl, and amino functional groups. The heteroatoms of the antibody's nucleophilic group are reactive with the electrophilic group on the elongation unit precursor and provide a covalent bond between the ligand unit and the elongation unit of the linker unit or drug-linker moiety. Electrophilic groups available for this purpose include, but are not limited to, maleimide, haloacetamide groups, and NHS esters. The electrophilic group provides a convenient site for antibody linking to form camptothecin conjugates or ligand unit-linker intermediates.

[0306] In other embodiments, the extension unit precursor has a reactive site with a nucleophilic group that reacts with an electrophilic group present on the ligand unit (e.g., antibody). Electrophilic groups available on the antibody for this purpose include, but are not limited to, aldehyde and ketone carbonyl groups. The heteroatoms of the nucleophilic group on the extension unit precursor can react with the electrophilic group on the antibody and form a covalent bond with the antibody. Nucleophilic groups available on the extension unit precursor for this purpose include, but are not limited to, acylhydrazides, hydroxylamines, amino groups, hydrazides, thioureas, carboxylic acid hydrazides, and aryl acylhydrazides. The electrophilic group on the antibody provides a convenient site for antibody linkage to form a camptothecin conjugate or a ligand unit-linker intermediate.

[0307] In some embodiments, the sulfur atom of the ligand unit is bonded to the succinimide ring system formed by the reaction of the thiol functional group of the targeting ligand with the maleimide moiety of the corresponding extension unit precursor. In other embodiments, the thiol functional group of the ligand unit reacts with the α-haloacetamide moiety to provide a sulfur-bonded extension unit via nucleophilic substitution of its halogen substituent.

[0308] Representative extended sub-units of this implementation scheme include those with the following structures:

[0309]

[0310] Among them, R 17 Adjacent wavy lines indicate parallel connector unit (B) or connector unit (A) (if B does not exist) or dispenser (S). * (If B does not exist) the connection, another wavy line indicates covalent connection with the sulfur atom of the ligand unit, and R 17 -C1-C 10 Alkylene-, C1-C 10 Heteroalkyl-, -C3-C8 carbocyclo-, -O-(C1-C8 alkylene)-, -arylene-, -C1-C 10 alkylene-arylene-, -arylene-C1-C 10 Alkylene-, -C1-C 10 Alkylene-(C3-C8 carbocyclic)-, -(C3-C8 carbocyclic)-C1-C 10 alkylene-, -C3-C8 heterocyclic-, -C1-C 10 alkylene-(C3-C8 heterocyclic)-, -(C3-C8 heterocyclic)-C1-C 10 Alkylene-, -C1-C 10 Alkylene C(=O)-, C1-C 10 Heteroalkyl-C(=O)-, -C3-C8 carbocyclic-C(=O)-, -O-(C1-C8 alkylene)-C(=O)-, -arylene-C(=O)-, -C1-C 10 alkylene-arylene-C(=O)-, -arylene-C1-C 10 Alkylene -C(=O)-, -C1-C 10 Alkylene-(C3-C8 carbocyclic)-C(=O)-, -(C3-C8 carbocyclic)-C1-C 10 Alkylene-C(=O)-, -C3-C8 heterocyclic-C(=O)-, -C1-C 10 Alkylene-(C3-C8 heterocyclic)-C(=O)-, -(C3-C8 heterocyclic)-C1-C 10Alkylene -C(=O)-, -C1-C 10 Alkylene-NH-, C1-C 10 Heteroalkyl-NH-, -C3-C8 carbocyclic-NH-, -O-(C1-C8 alkylene)-NH-, -arylene-NH-, -C1-C 10 alkylene-arylene-NH-, -arylene-C1-C 10 Alkylene -NH-, -C1-C 10 Alkylene-(C3-C8 carbocyclic)-NH-, -(C3-C8 carbocyclic)-C1-C 10 Alkylene-NH-, -C3-C8 heterocyclic-NH-, -C1-C 10 alkylene-(C3-C8 heterocyclic)-NH-, -(C3-C8 heterocyclic)-C1-C 10 Alkylene -NH-, -C1-C 10 Alkylene-S-, C1-C 10 Heteroalkyl-S-, -C3-C8 carbide-S-, -O-(C1-C8 alkylene)-S-, -arylene-S-, -C1-C 10 alkylene-arylene-S-, -arylene-C1-C 10 Alkylene -S-, -C1-C 10 Alkylene-(C3-C8 carbocyclic)-S-, -(C3-C8 carbocyclic)-C1-C 10 Alkylene-S-, -C3-C8 heterocyclic-S-, -C1-C 10 alkylene-(C3-C8 heterocyclic)-S- or -(C3-C8 heterocyclic)-C1-C 10 Alkylene-S-.

[0311] In some implementation schemes, R 17 The group is optionally surrounded by a basic unit (BU), such as an aminoalkyl moiety, for example –(CH2). x NH2、–(CH2) x NHR a and –(CH2) x NR a Replaced by 2, where the subscript x is an integer from 1 to 4 and each R a Selected independently from C 1-6 Alkyl and C 1-6 Halogenated alkyl, or two R a The groups combine with the nitrogen atoms to which they are attached to form nitrogen heterocyclic butyl, pyrrolidinyl, or piperidinyl groups.

[0312] An illustrative extended subunit is an extended subunit of formula Za or Za-BU, where R 17 -C1-C10 Alkylene -C(=O)-, -C1-C 10 Heteroalkyl-C(=O)-, -C3-C8 carbocyclic-C(=O)-, -O-(C1-C8 alkylene)-C(=O)-, -arylene-C(=O)-, -C1-C 10 alkylene-arylene-C(=O)-, -arylene-C1-C 10 Alkylene -C(=O)-, -C1-C 10 Alkylene-(C3-C8 carbocyclic)-C(=O)-, -(C3-C8 carbocyclic)-C1-C 10 Alkylene-C(=O)-, -C3-C8 heterocyclic-C(=O)-, -C1-C 10 alkylene-(C3-C8 heterocyclic)-C(=O)- or -(C3-C8 heterocyclic)-C1-C 10 Alkylene-C(=O)-.

[0313] Therefore, some preferred embodiments are represented by equations Za and Za-BU:

[0314]

[0315] The wavy line adjacent to the carbonyl carbon atom indicates a connection to B, A, or S* in the above formula (depending on the presence of A and / or B), while the other wavy line indicates a covalent bond between the succinimide ring carbon atom and the sulfur atom of the ligand unit. During synthesis, the basic amino functional group of the basic unit (BU) can be protected by a protecting group.

[0316] A more preferred embodiment of the extended subunits of Za and Za-BU is as follows:

[0317]

[0318] The wavy line adjacent to the carbonyl carbon atom indicates the connection with B, A, or S* in the above formula (depending on the presence of A and / or B), and the other wavy line indicates the covalent bond between the carbon atom of the succinimide ring and the sulfur atom of the ligand unit.

[0319] It will be understood that ligand-substituted succinimides can exist in one or more hydrolytic forms. The following examples of those forms are illustrated with respect to the hydrolysis of Za or Za-BU, wherein the structures representing the regioisomers derived from this hydrolysis have the formulas Zb and Zc or Zb-BU and Zc-BU.

[0320] Therefore, in other preferred embodiments, the extended subunit (Z) is composed of the succinic acid-amide portion as shown below:

[0321]

[0322]

[0323] This depends on the presence of A and / or B, and the bonding to R. 17 The wavy lines adjacent to the carbonyl carbon atom and the wavy lines adjacent to the carbon atom of the acid-amide moiety are as defined for Za or Za-BU; and R 17 It is –C1-C5 alkylene-, wherein in Zb-BU and Zc-BU the alkylene is replaced by a basic unit (BU), wherein BU is –(CH2). x NH2、–(CH2) x NHR a Or –(CH2) x N(R a )2, where the subscript x is an integer from 1 to 4 and each R a Selected independently from C 1-6 Alkyl and C 1-6 Halogenated alkyl, or two R a Together with the nitrogen to which they are attached, they define a nitrogen heterocyclic butyl, pyrrolidinyl, or piperidinyl group.

[0324] In a more preferred embodiment, -ZA- comprises a portion derived from the maleimide-alkanoic acid moiety or the mDPR moiety. See, for example, WO 2013 / 173337. In one set of embodiments, ZA- is derived from the maleimide-propionyl moiety.

[0325] Therefore, in some preferred embodiments, the extended subunit (Z) is composed of a succinic acid-amide moiety represented by the following structures: Zb', Zc', (R / S)-Zb'-BU, (S)-Zb'-BU, (R / S)-Zc'-BU, or (S)-Zc'-BU:

[0326]

[0327] The wavy lines are defined as for Za or Za-BU.

[0328] In a particularly preferred embodiment, the extended subunit (Z) consists of a succinimide moiety represented by the following structure:

[0329]

[0330] Or it may consist of a succinic acid-amide moiety represented by the following structure:

[0331]

[0332] An exemplary extension subunit bonded to the connector unit (A) is composed of Za', Zb', or Zc', wherein the –R of Za, Zb, or Zc17 It is –CH2- or -CH2CH2-, or composed of Za'-BU, Zb'-BU, or Zc'-BU, where the –R of Za-BU, Zb-BU, or Zc-BU is... 17 (BU)- is –CH(CH2NH2)- and has the following structure:

[0333]

[0334] The wavy lines are defined as for Za or Za-BU.

[0335] Other extended subunits bonded to the ligand unit (L) and the connector unit (A) have the above-described structures, wherein the A in any of the above-described -Za-A-, -Za(BU)-A-, -Za'-A-, -Za'(BU)-A-, -Zb-A-, -Zb(BU)-A-, -Zb'-A-, -Zb'(BU)-, -Zc'-A-, and Zc'(BU)-A- structures is replaced by a parallel connector unit having the following structure:

[0336]

[0337] Where the subscript n is 8-24; R PEG The terminator is the PEG unit end capping group, preferably -CH3 or –CH2CH2CO2H. An asterisk (*) indicates a covalent connection with the extended subunit corresponding to the formula Za, Za', Zb' or Zc', and a wavy line indicates a covalent connection with the releasable linker (RL).

[0338] An exemplary extended subunit (i.e., the extended subunit precursor) prior to conjugation with the ligand unit is composed of a maleimide moiety and is represented by a structure including formula Z'a:

[0339]

[0340] The wavy line adjacent to the carbonyl carbon atom indicates a connection to B, A, or S* in the above formula (depending on the presence of A and / or B); R 17 For –(CH2) 1-5 - Optional basic unit such as an optional substituted aminoalkyl group, for example –(CH2). x NH2、–(CH2) x NHR a and –(CH2) x N(R a Replaced by 2, where the subscript x is an integer from 1 to 4 and each R a Selected independently from C 1-6 Alkyl and C 1-6 Halogenated alkyl, or two R aThe groups combine with the nitrogen atoms to which they are attached to form nitrogen heterocyclic butyl, pyrrolidinyl, or piperidinyl groups.

[0341] The other illustrative extended subunits (i.e., extended subunit precursors) preceding the ligand unit are composed of maleimide moieties and are represented by structures including the formula Z'a-BU.

[0342]

[0343] The wavy line adjacent to the carbonyl carbon atom indicates a connection to B, A, or S* in the above formula (depending on the presence of A and / or B), R 17 For –(CH2) 1-5 - Optional basic unit such as an optional substituted aminoalkyl group, for example –(CH2). x NH2、–(CH2) x NHR a and –(CH2) x N(R a Replaced by )2, where the subscript x is an integer from 1 to 4, preferably R. 17 It is -CH2- or -CH2CH2- and the subscript x is 1 or 2, and each R a Selected independently from C 1-6 Alkyl and C 1-6 Halogenated alkyl, or two R a The groups combine with the nitrogen atoms to which they are attached to form nitrogen heterocyclic butyl, pyrrolidinyl, or piperidinyl groups.

[0344] In some preferred embodiments of formula Z'a, the extended subunit precursor is represented by one of the following structures:

[0345]

[0346] The wavy line adjacent to the carbonyl group is as defined for Z'a or Z'a-BU.

[0347] In a more preferred embodiment, the extended subunit precursor (Z') is composed of a maleimide moiety and is represented by the following structure:

[0348]

[0349] The wavy line adjacent to the carbonyl group is as defined for Za' and the amino group is optionally protonated or protected by an amino protecting group.

[0350] In extended subunits with the BU moiety, it should be understood that during synthesis, the amino functional groups of this moiety are typically protected by amino protecting groups such as acid-instable protecting groups (e.g., BOC).

[0351] With the joint unit composed of structures from Za' or Z'a-BU (where –R 17 -or–R 17 (BU)- is a schematic extended subunit precursor covalently linked to –CH2-, -CH2CH2-, or –CH(CH2NH2)-, and has the following structure:

[0352]

[0353] The wavy line adjacent to the carbonyl group is as defined for Z'a or Z'a-BU.

[0354] Other extended subunit precursors bonded to the connector unit (A) have the above-described structure, wherein the A in either of the Z'-A- and Z'(BU)-A- structures is replaced by a parallel connector unit and a dispensing agent (-B(S*)-) having the following structure:

[0355]

[0356] Where the subscript n is in the range of 8 to 24; R PEG The terminator is the PEG unit end capping group, preferably –CH3 or –CH2CH2CO2H. An asterisk (*) indicates a covalent connection to the extended subunit monomer corresponding to the formula Za or Za', and a wavy line indicates a covalent connection to RL. In those cases shown here, the PEG group shown is an example of various dispensing agents, including PEG groups of different lengths and other dispensing agents that can be directly attached or modified to attach to parallel connector units.

[0357] In another embodiment, the extended subunit is connected to the ligand unit via a disulfide bond between the sulfur atom of the ligand unit and the sulfur atom of the extended subunit. A representative extended subunit of this embodiment is depicted within square brackets of formula Zb:

[0358]

[0359] The wavy line indicates a parallel connector unit (B) or connector unit (A) (if B does not exist) or dispenser (S). * (If A and B do not exist) connection, and R 17 -C1-C 10 Alkylene-, C1-C 10 Heteroalkyl-, -C3-C8 carbocyclo-, -O-(C1-C8 alkylene)-, -arylene-, -C1-C 10 alkylene-arylene-, -arylene-C1-C 10 Alkylene-, -C1-C 10 Alkylene-(C3-C8 carbocyclic)-, -(C3-C8 carbocyclic)-C1-C10 alkylene-, -C3-C8 heterocyclic-, -C1-C 10 alkylene-(C3-C8 heterocyclic)-, -(C3-C8 heterocyclic)-C1-C 10 Alkylene-, -C1-C 10 Alkylene C(=O)-, C1-C 10 Heteroalkyl-C(=O)-, -C3-C8 carbocyclic-C(=O)-, -O-(C1-C8 alkylene)-C(=O)-, -arylene-C(=O)-, -C1-C 10 alkylene-arylene-C(=O)-, -arylene-C1-C 10 Alkylene -C(=O)-, -C1-C 10 Alkylene-(C3-C8 carbocyclic)-C(=O)-, -(C3-C8 carbocyclic)-C1-C 10 Alkylene-C(=O)-, -C3-C8 heterocyclic-C(=O)-, -C1-C 10 Alkylene-(C3-C8 heterocyclic)-C(=O)-, -(C3-C8 heterocyclic)-C1-C 10 Alkylene -C(=O)-, -C1-C 10 Alkylene-NH-, C1-C 10 Heteroalkyl-NH-, -C3-C8 carbocyclic-NH-, -O-(C1-C8 alkylene)-NH-, -arylene-NH-, -C1-C 10 alkylene-arylene-NH-, -arylene-C1-C 10 Alkylene -NH-, -C1-C 10 Alkylene-(C3-C8 carbocyclic)-NH-, -(C3-C8 carbocyclic)-C1-C 10 Alkylene-NH-, -C3-C8 heterocyclic-NH-, -C1-C 10 alkylene-(C3-C8 heterocyclic)-NH-, -(C3-C8 heterocyclic)-C1-C 10 Alkylene -NH-, -C1-C 10 Alkylene-S-, C1-C 10 Heteroalkyl-S-, -C3-C8 carbocyclo-S-, -O-(C1-C8 alkylene)-S-, -arylene-S-, -C1-C 10 alkylene-arylene-S-, -arylene-C1-C 10 Alkylene -S-, -C1-C 10 Alkylene-(C3-C8 carbocyclic)-S-, -(C3-C8 carbocyclic)-C1-C 10 Alkylene-S-, -C3-C8 heterocyclic-S-, -C1-C10 alkylene-(C3-C8 heterocyclic)-S- or -(C3-C8 heterocyclic)-C1-C 10 Alkylene-S-.

[0360] In yet another embodiment, the reactive group of the extended subunit precursor contains a reactive site that can form a bond with a primary or secondary amino group of the ligand unit. Examples of such reactive sites include, but are not limited to, activated esters such as succinimidyl esters, 4-nitrophenyl esters, pentafluorophenyl esters, tetrafluorophenyl esters, acid anhydrides, acyl chlorides, sulfonyl chlorides, isocyanates, and isothiocyanates. Representative extended subunits of this embodiment are depicted in square brackets of formulas Zci, Zcii, and Zciii:

[0361]

[0362] The wavy line indicates a parallel connector unit (B) or connector unit (A) (if B does not exist) or dispenser (S). * (If A and B do not exist) the connection, and R 17 -C1-C 10 Alkylene-, C1-C 10 Heteroalkyl-, -C3-C8 carbocyclo-, -O-(C1-C8 alkylene)-, -arylene-, -C1-C 10 alkylene-arylene-, -arylene-C1-C 10 Alkylene-, -C1-C 10 Alkylene-(C3-C8 carbocyclic)-, -(C3-C8 carbocyclic)-C1-C 10 alkylene-, -C3-C8 heterocyclic-, -C1-C 10 alkylene-(C3-C8 heterocyclic)-, -(C3-C8 heterocyclic)-C1-C 10 Alkylene-, -C1-C 10 Alkylene-C(=O)-, C1-C 10 Heteroalkyl-C(=O)-, -C3-C8 carbocyclic-C(=O)-, -O-(C1-C8 alkylene)-C(=O)-, -arylene-C(=O)-, -C1-C 10 alkylene-arylene-C(=O)-, -arylene-C1-C 10 Alkylene -C(=O)-, -C1-C 10 Alkylene-(C3-C8 carbocyclic)-C(=O)-, -(C3-C8 carbocyclic)-C1-C 10 Alkylene-C(=O)-, -C3-C8 heterocyclic-C(=O)-, -C1-C 10Alkylene-(C3-C8 heterocyclic)-C(=O)-, -(C3-C8 heterocyclic)-C1-C 10 Alkylene -C(=O)-, -C1-C 10 Alkylene-NH-, C1-C 10 Heteroalkyl-NH-, -C3-C8 carbocyclic-NH-, -O-(C1-C8 alkylene)-NH-, -arylene-NH-, -C1-C 10 alkylene-arylene-NH-, -arylene-C1-C 10 Alkylene -NH-, -C1-C 10 Alkylene-(C3-C8 carbocyclic)-NH-, -(C3-C8 carbocyclic)-C1-C 10 Alkylene-NH-, -C3-C8 heterocyclic-NH-, -C1-C 10 alkylene-(C3-C8 heterocyclic)-NH-, -(C3-C8 heterocyclic)-C1-C 10 Alkylene -NH-, -C1-C 10 Alkylene-S-, C1-C 10 Heteroalkyl-S-, -C3-C8 carbocyclo-S-, -O-(C1-C8 alkylene)-S-, -arylene-S-, -C1-C 10 alkylene-arylene-S-, -arylene-C1-C 10 Alkylene -S-, -C1-C 10 Alkylene-(C3-C8 carbocyclic)-S-, -(C3-C8 carbocyclic)-C1-C 10 Alkylene-S-, -C3-C8 heterocyclic-S-, -C1-C 10 alkylene-(C3-C8 heterocyclic)-S- or -(C3-C8 heterocyclic)-C1-C 10 Alkylene-S-.

[0363] In other embodiments, the reactive group of the extended subunit precursor contains a reactive nucleophile capable of reacting with an electrophile present on or introduced onto the ligand unit. For example, the carbohydrate moiety on the targeting ligand can be gently oxidized using a reagent such as sodium periodate, and the electrophilic functional group (-CHO) generated by the oxidized carbohydrate can condense with the extended subunit precursor containing the reactive nucleophile, such as acylhydrazides, oximes, primary or secondary amines, hydrazides, thioureas, carboxylic acid hydrazides, or aryl acylhydrazides, as described in Kaneko, T. et al. (1991) Bioconjugate Chem. 2:133-41. Representative extended subunits of this embodiment are depicted in square brackets of formulas Zdi, Zdii, and Zdiii:

[0364]

[0365]

[0366] The wavy line indicates a parallel connector unit (B) or connector unit (A) or dispensing agent (S). * (If A and B do not exist) the connection, and R 17 -C1-C 10 Alkylene-, C1-C 10 Heteroalkyl-, -C3-C8 carbocyclo-, -O-(C1-C8 alkylene)-, -arylene-, -C1-C 10 alkylene-arylene-, -arylene-C1-C 10 Alkylene-, -C1-C 10 Alkylene-(C3-C8 carbocyclic)-, -(C3-C8 carbocyclic)-C1-C 10 alkylene-, -C3-C8 heterocyclic-, -C1-C 10 alkylene-(C3-C8 heterocyclic)-, -(C3-C8 heterocyclic)-C1-C 10 Alkylene-, -C1-C 10 Alkylene-C(=O)-, C1-C 10 Heteroalkyl-C(=O)-, -C3-C8 carbocyclic-C(=O)-, -O-(C1-C8 alkylene)-C(=O)-, -arylene-C(=O)-, -C1-C 10 alkylene-arylene-C(=O)-, -arylene-C1-C 10 Alkylene -C(=O)-, -C1-C 10 Alkylene-(C3-C8 carbocyclic)-C(=O)-, -(C3-C8 carbocyclic)-C1-C 10 Alkylene-C(=O)-, -C3-C8 heterocyclic-C(=O)-, -C1-C 10 Alkylene-(C3-C8 heterocyclic)-C(=O)-, -(C3-C8 heterocyclic)-C1-C 10 Alkylene -C(=O)-, -C1-C 10 Alkylene-NH-, C1-C 10 Heteroalkyl-NH-, -C3-C8 carbocyclic-NH-, -O-(C1-C8 alkylene)-NH-, -arylene-NH-, -C1-C 10 alkylene-arylene-NH-, -arylene-C1-C 10 Alkylene -NH-, -C1-C 10 Alkylene-(C3-C8 carbocyclic)-NH-, -(C3-C8 carbocyclic)-C1-C10 Alkylene-NH-, -C3-C8 heterocyclic-NH-, -C1-C 10 alkylene-(C3-C8 heterocyclic)-NH-, -(C3-C8 heterocyclic)-C1-C 10 Alkylene -NH-, -C1-C 10 Alkylene-S-, C1-C 10 Heteroalkyl-S-, -C3-C8 carbocyclo-S-, -O-(C1-C8 alkylene)-S-, -arylene-S-, -C1-C 10 alkylene-arylene-S-, -arylene-C1-C 10 Alkylene -S-, -C1-C 10 Alkylene-(C3-C8 carbocyclic)-S-, -(C3-C8 carbocyclic)-C1-C 10 Alkylene-S-, -C3-C8 heterocyclic-S-, -C1-C 10 alkylene-(C3-C8 heterocyclic)-S- or -(C3-C8 heterocyclic)-C1-C 10 Alkylene-S-.

[0367] In some aspects of the invention, the mass of the extended subunit is not more than about 1,000 Daltons, not more than about 500 Daltons, not more than about 200 Daltons, is about 30, 50 or 100 Daltons to about 1,000 Daltons, is about 30, 50 or 100 Daltons to about 500 Daltons, or is about 30, 50 or 100 Daltons to about 200 Daltons.

[0368] Connector unit (A)

[0369] In some embodiments, the camptothecin conjugate or camptothecin-linker compound contains a linker unit (A) to add additional distance between the extended subunit (Z) or its precursor (Z') and the releasable linker, if required. In some embodiments, the additional distance facilitates activation within the RL. Accordingly, when present, the linker unit (A) extends the framework of the linker unit. At this point, the linker unit (A) is covalently bonded at one end to the extended subunit (or its precursor) and at its other end to an optional parallel linker unit or dispensing agent (S*).

[0370] Those skilled in the art will understand that the linker unit can be any group used to provide a releasable connection between the linker and the remainder of the linker unit (Q). The linker unit can, for example, consist of one or more (e.g., 1-10, preferably 1, 2, 3, or 4) natural or non-natural amino acids, amino alcohols, amino aldehydes, or diamino residues. In some embodiments, the linker unit is a single natural or non-natural amino acid, amino alcohol, amino aldehyde, or diamino residue. An exemplary amino acid that can serve as a linker unit is β-alanine.

[0371] In some of those implementations, the connector unit has the following formula:

[0372]

[0373]

[0374] The wavy line represents the connection between the camptothecin conjugate or camptothecin linker compound and the linker unit; and R... 111 Independently selected from hydrogen, p-hydroxybenzyl, methyl, isopropyl, isobutyl, sec-butyl, -CH2OH, -CH(OH)CH3, -CH2CH2SCH3, -CH2CONH2, -CH2COOH, -CH2CH2CONH2, -CH2CH2COOH, -(CH2)3NHC(=NH)NH2, -(CH2)3NH2, -(CH2)3NHCOCH3, -(CH2)3NHCHO, -(CH2)4NHC(=NH)NH2, -(CH2)4NH2, -(CH2)4NHCOCH3, -(CH2)4NHCHO, -(CH2)3NHCONH2, -(CH2)4NHCONH2, -CH2CH2CH(OH)CH2NH2, 2-pyridylmethyl-, 3-pyridylmethyl-, 4-pyridylmethyl-,

[0375]

[0376] And each R 100 The components are independently selected from hydrogen or -C1-C3 alkyl groups, preferably hydrogen or CH3; and the subscript c is an integer independently selected from 1 to 10, preferably 1 to 3.

[0377] It has a carbonyl group so as to react with the dispensing agent (S) * ) or with –B(S * A representative connector unit for this type of connection is as follows:

[0378]

[0379] In each case, R 13Independently selected from -C1-C6 alkylene-, -C3-C8 carbocyclo-, -arylene-, -C1-C 10 Heteroalkyl-, -C3-C8 heterocyclic-, -C1-C 10 alkylene-arylene-, -arylene-C1-C 10 Alkylene-, -C1-C 10 Alkylene-(C3-C8 carbocyclic)-, -(C3-C8 carbocyclic)-C1-C 10 Alkylene-, -C1-C 10 alkylene-(C3-C8 heterocyclic)- and -(C3-C8 heterocyclic)-C1-C 10 Alkylene, and the subscript c is an integer in the range of 1 to 4. In some embodiments, R 13 It is a -C1-C6 alkylene group, and c is 1.

[0380] It has a carbonyl group so as to react with the dispensing agent (S) * ) or with –B(S * Another representative connector unit for the )- connection is as follows:

[0381]

[0382] Where R 13 It is -C1-C6 alkylene-, -C3-C8 carbocyclo-, -arylene-, -C1-C 10 Heteroalkyl-, -C3-C8 heterocyclic-, -C1-C 10 alkylene-arylene-, -arylene-C1-C 10 Alkylene-, -C1-C 10 Alkylene-(C3-C8 carbocyclic)-, -(C3-C8 carbocyclic)-C1-C 10 Alkylene-, -C1-C 10 alkylene-(C3-C8 heterocyclic)-, or -(C3-C8 heterocyclic)-C1-C 10 Alkylene-. In some embodiments, R 13 It is a -C1-C6 alkylene group.

[0383] With dispensing agent (S) * ) or with –B(S * A representative connector unit for the NH section of the connection is as follows:

[0384]

[0385] In each case, R 13 Independently selected from -C1-C6 alkylene-, -C3-C8 carbocyclo-, -arylene-, -C1-C10 Heteroalkyl-, -C3-C8 heterocyclic-, -C1-C 10 alkylene-arylene-, -arylene C1-C 10 Alkylene-, -C1-C 10 Alkylene-(C3-C8 carbocyclic)-, -(C3-C8 carbocyclic)-C1-C 10 Alkylene-, -C1-C 10 alkylene-(C3-C8 heterocyclic)- and -(C3-C8 heterocyclic)-C1-C 10 Alkylene-, and the subscript c is 1 to 14. In some embodiments, R 13 It is a -C1-C6 alkylene group, and the subscript c is 1.

[0386] With dispensing agent (S) * ) or with –B(S * Another representative connector unit for the NH section of the connection is as follows:

[0387]

[0388] Where R 13 It is -C1-C6 alkylene-, -C3-C8 carbocyclo-, -arylene-, -C1-C 10 Heteroalkyl-, -C3-C8 heterocyclic-, -C1-C 10 alkylene-arylene-, -arylene-C1-C 10 Alkylene-, -C1-C 10 Alkylene-(C3-C8 carbocyclic)-, -(C3-C8 carbocyclic)-C1-C 10 Alkylene-, -C1-C 10 alkylene-(C3-C8 heterocyclic)-, -(C3-C8 heterocyclic)-C1-C 10 Alkylene-, –C(=O)C1-C6 alkylene-, or -C1-C6 alkylene-C(=O)-C1-C6 alkylene.

[0389] Selected implementations of the connector unit include those with the following structures:

[0390]

[0391] The wavy line adjacent to nitrogen indicates a covalent bond with the extended subunit (Z) (or its precursor Z'), and the wavy line adjacent to the carbonyl group indicates a bond with the partitioning agent (S). * ) or with –B(S * The covalent connection of )-; and m is an integer in the range of 1 to 6, preferably 2 to 6, more preferably 2 to 4.

[0392] Releasable Connector (RL):

[0393] The glucuronide unit is a releasable linker that provides a mechanism for the separation of camptothecin from the ligand unit and other components of the linker unit by activating a self-elimination cascade reaction within the linker unit. In this embodiment, the self-elimination cascade reaction is activated by manipulating a glycosidase on the carbohydrate moiety of the glucuronide unit. Many sugars can be used in the embodiments described herein. Specific carbohydrate moieties include those of galactose, glucose, mannose, xylose, arabinose, mannose-6-phosphate, fucose, rhamnose, fructose, allose, 6-deoxyglucose, lactose, maltose, cellobiose, gentiobiose, maltotriose, GlcNAc, GalNAc, and maltohexasose.

[0394] A glycosidic unit typically comprises a sugar moiety (Su) linked to a self-eliminating spacer via an oxyglycosidic bond. Cleavage of the oxyglycosidic bond initiates a self-elimination sequence, resulting in the release of a free drug. In some embodiments, the self-elimination sequence is activated by the cleavage of a glucuronide unit, which is an exemplary glycosidic unit, by a β-glucuronidase. The glucuronide unit comprises an activation unit and a self-eliminating spacer unit. The glucuronide unit comprises a sugar moiety (Su) linked to the self-eliminating spacer unit via an oxyglycosidic bond.

[0395] In some embodiments, the glucuronide unit comprises a sugar moiety (Su) linked via an oxyglycosidic bond (-O'-) to a self-eliminating unit (SP) of the following formula:

[0396]

[0397] The wavy line represents a drug unit covalently connected to any one of formulas CPT1, CPT2, CPT3, CPT4, CPT5, CPT6, and CPT7, depending on the specific circumstances, or a spacer subunit covalently connected to a drug unit (camptothecin compound), and a dispensing agent (S) via a connector unit (A) or a parallel connector unit (B). * The joint unit or a combination of the joint unit and the parallel joint unit are directly or indirectly covalently connected to the extension subunit (Z) or its precursor (Z').

[0398] O-glycosidic bonds (-O'-) are typically cleavage sites of β-glucuronidase (i.e., Su comes from glucuronic acid), such as glycosidic bonds that can be cleaved by human lysosomal β-glucuronidase.

[0399] In some implementations, the glucuronide unit may be represented by the formula Ga or Gb:

[0400]

[0401] Where Su is the sugar moiety, and -O'- represents an oxyglycosidic bond; R1S R 2S and R 3S Independently, it is hydrogen, halogen, -CN, -NO2 or other electron-withdrawing or electron-donating groups; and wherein the wavy line indicates a connection to the extended subunit (Z) (or its precursor (Z'), directly or indirectly through a linker unit or parallel linker unit or linker unit and parallel linker unit); and # indicates a connection to camptothecin or a spacer (directly or indirectly through an intermediate functional group or other part).

[0402] In the preferred embodiment, R 1S R 2S and R 3S Independently selected from hydrogen, halogen, -CN, or -NO2. In other preferred embodiments, R 1S R 2S and R 3S Each is hydrogen. In other preferred embodiments, R 2S It is an electron-withdrawing group, preferably NO2, and R 1S and R 3S Each is hydrogen.

[0403] In some of these respects, the activatable self-eliminating group capable of being cleaved by glycosidases to initiate a self-elimination reaction sequence is represented by the formula Gc:

[0404]

[0405] Where R 4S CH2OH or –CO2H, the wavy line indicates direct or indirect covalent connection to the extended subunit (Z) (or its precursor Z') through a joint unit or parallel joint unit or joint unit and parallel joint unit, and the hash (#) indicates covalent connection to the methylene carbamate unit.

[0406] In some embodiments in which the activatable self-eliminating portion is composed of glucuronide units, it is represented by the following formula:

[0407]

[0408] The wavy line indicates direct or indirect covalent connection to the extended subunit (Z) (or its precursor Z') via a joint unit or parallel joint unit or joint unit and parallel joint unit, and the hash symbol (#) indicates a benzyl carbon covalently connected to a spacer or functional group connected to camptothecin.

[0409] Another type of releasable linker, which provides a mechanism for separating camptothecin from the ligand unit and other components of the linker unit by activating a self-elimination cascade reaction within the linker unit, consists of a p-aminobenzyloxycarbonyl (PAB) moiety, whose phenylene component is... mSubstitution, wherein the subscript m indicating the number of substituents is an integer from 0 to 4, and each J is independently -C1-C8 alkyl, -O-(C1-C8 alkyl), -halogen, -nitro, or -cyano.

[0410] In some embodiments, RL is a self-eliminating group capable of releasing -D without a separate hydrolysis step or subsequent self-elimination event. In some embodiments, -RL- is a PAB moiety linked to the carbonyl group of -W- via the amino nitrogen atom of the PAB group and directly linked to the PAB portion of -D via a carbonate group. In related embodiments, -RL- is composed of a PAB moiety that is linked to -A- and -S via the amino nitrogen atom of the PAB group. * - or -B- carbonyl linkage, and directly linked to -D via a carbonate group. Not bound by any particular theory or mechanism, Toki et al. (2002) J Org. Chem. 67: 1866-1872 showed a possible mechanism for drug release from RL composed of a PAB moiety, in which RL is directly linked to -D via a carbonate group.

[0411] In some implementations, the RL element containing the PAB portion is represented by the following formula:

[0412]

[0413] The subscript m is an integer from 0 to 4, and each J is independently -C1-C8 alkyl, -O-(C1-C8 alkyl), -halogen, -nitro or -cyano.

[0414] Other examples of self-eliminating groups include, but are not limited to, electronically similar to the PAB moiety, such as 2-aminoimidazolium-5-methanol derivatives (Hay et al. (1999) Bioorg. Med. Chem. Lett. 9: 2237) and aromatic compounds of ortho- or p-aminobenzyl acetals. Other RLs undergo cyclization upon hydrolysis of the amide bond, such as substituted and unsubstituted 4-aminobutyric amides (Rodrigues et al., Chemistry Biology, 1995, 2, 223), appropriately substituted bicyclic [2.2.1] and bicyclic [2.2.2] cyclic systems (Storm et al., J. Amer. Chem. Soc., 1972, 94, 5815) and 2-aminophenylpropionic amides (Amsberry et al., J. Org. Chem., 1990, 55, 5867).

[0415] In one implementation, RL is a branched bis(hydroxymethyl)styrene (BHMS) unit.

[0416] In some implementations, RL has the following formula:

[0417]

[0418] The wavy lines marked with ** indicate the connection sites with D; and the wavy lines marked with * indicate the connection sites with other connector components of Q.

[0419] In some embodiments, the linker portion comprises a heterocyclic "self-eliminating moiety" of formula I, II, or III that binds to the drug and incorporates an amide group that is initiated by hydrolysis by an intracellular protease, the reaction ultimately cleaving the self-eliminating moiety from the drug, thereby releasing the drug from the conjugate in its active form. The linker portion also comprises a peptide sequence adjacent to the self-eliminating moiety, which is a substrate of an intracellular enzyme such as a cathepsin (e.g., cathepsin B), which cleaves the peptide at an amide bond shared with the self-eliminating moiety. For the embodiments disclosed herein, the PAB-containing RL is directly linked to a tertiary hydroxyl group of the lactone ring present in each of CPT1-CPT7, each of compounds 14-14z in Table I, or each of compounds 18a-18r in Table J.

[0420] In some embodiments, the heterocyclic self-eliminating group (RL) is selected from formulas I, II, and III:

[0421]

[0422] The wavy line represents the covalently linked site with the cell-specific ligand and the drug moiety, where U represents O, S, or NR. 6 Q is CR 4 Or N; V 1 V 2 and V 3 Independent for CR 4 Or N, provided that for equations II and III, Q and V 1 and V 2 At least one of them is N; T is O that is suspended from CPT1, CPT2, CPT3, CPT4, CPT5, CPT6 or CPT7;

[0423] R 1 R 2 R 3 and R 4 Independently selected from H, F, Cl, Br, I, OH, -N(R) 5 )2、-N(R 5 )3 + C1-C8 alkyl halides, carboxylic acid esters, sulfate esters, aminosulfonates, sulfonates, -SO2R 5 -S(=O)R 5 -SR 5 -SO2N(R) 5 )2、-C(=O)R5 -CO2R 5 -C(=O)N(R) 5 )2, -CN, -N3, -NO2, C1-C8 alkoxy, C1-C8 haloalkyl, polyvinyloxy, phosphonate, phosphate, C1-C8 alkyl, C1-C8 substituted alkyl, C2-C8 alkenyl, C2-C8 substituted alkenyl, C2-C8 alkynyl, C2-C8 substituted alkynyl, C6-C 20 Aryl, C6-C 20 Substituted aryl, C1-C 20 Heterocyclic rings and C1-C 20 Substitute heterocyclic rings; or R 2 and R 3 Together they form a carbonyl group (=O) or a spirocarbocyclic ring with 3-7 carbon atoms; and

[0424] R 5 and R 6 Independently selected from H, C1-C8 alkyl, C1-C8 substituted alkyl, C2-C8 alkenyl, C2-C8 substituted alkenyl, C2-C8 alkynyl, C2-C8 substituted alkynyl, C6-C 20 Aryl, C6-C 20 Substituted aryl, C1-C 20 Heterocyclic rings and C1-C 20 Replacement of heterocyclic rings;

[0425] Among them, C1-C8 substituted alkyl, C2-C8 substituted alkenyl, C2-C8 substituted alkynyl, C6-C 20 Substituted aryl and C2-C 20 The substituted heterocycle is independently substituted by one or more substituents selected from the following: F, Cl, Br, I, OH, -N(R) 5 )2、-N(R 5 )3 + C1-C8 alkyl halides, carboxylic acid esters, sulfate esters, aminosulfonates, sulfonates, C1-C8 alkylsulfonates, C1-C8 alkylamino, 4-dialkylaminopyridine, C1-C8 alkylhydroxy, C1-C8 alkylthiols, -SO2R 5 -S(=O)R 5 -SR 5 -SO2N(R) 5 )2,-C(=O)R 5 ,-CO2R 5 -C(=O)N(R) 5 )2, -CN, -N3, -NO2, C1-C8 alkoxy, C1-C8 trifluoroalkyl, C1-C8 alkyl, C3-C 12 Carbon rings, C6-C 20 Aryl, C2-C 20Heterocyclic compounds, polyethylene oxides, phosphonates, and phosphate esters.

[0426] The conjugate is stable outside the cell or in the absence of enzymes capable of cleaving the amide bond of the self-eliminating moiety. However, upon entering the cell or being exposed to a suitable enzyme, the amide bond is cleaved, triggering a spontaneous self-elimination reaction that leads to the cleavage of the covalent bond linking the self-eliminating moiety to the drug, thereby releasing the drug in its underivative or pharmacologically active form.

[0427] The self-eliminating portion of the conjugates of the present invention is further incorporated with one or more heteroatoms, thereby providing improved solubility, increased cleavage rate, and / or reduced tendency for conjugate aggregation. In some cases, these improvements of the heterocyclic self-eliminating linker constructs of the present invention relative to non-heterocyclic PAB-type linkers lead to surprising and unexpected biological properties, such as increased efficacy, reduced toxicity, and / or improvements in one or more desired pharmacokinetic and / or pharmacodynamic properties.

[0428] It should be understood that T in Formulas I-III is O, because it is derived from the tertiary hydroxyl (-OH) on the lactone ring moiety of any of CPT1, CPT2, CPT3, CPT4, CPT5, CPT6, CPT7, compounds 14a-14z in Table I, and compounds 18a-18r in Table J.

[0429] Regardless of theory or any specific mechanism, the presence of electron-withdrawing groups on the heterocycles of Formula I, II, or III can sometimes reduce the cleavage rate.

[0430] In one embodiment, the self-eliminating moiety is a group of formula I, where Q is N and U is O or S. This group has a non-linear structural characteristic that improves the solubility of the conjugate. In this case, R is sometimes H, methyl, nitro, or CF3. In one embodiment, Q is N and U is O, thus forming an oxazole ring, and R is H. In another embodiment, Q is N and U is S, thus forming a thiazole ring optionally substituted with a Me or CF3 group at R.

[0431] In another exemplary embodiment, the self-eliminating moiety is a group of formula II, where Q is N and V 1 and V 2 Independently, it is N or CH. In another implementation, Q, V 1 and V 2 Each is N. In another implementation, Q and V 1 For N, and V 2 For CH. In another implementation, Q and V 2 For N, and V 1 For CH. In another implementation, Q and V 1 Both are CH, and V2 For N. In another implementation, Q is N, and V is N. 1 and V 2 All are CH.

[0432] In another embodiment, the self-eliminating moiety is a group of formula III, wherein Q, V 1 V 2 and V 3 Each can be N or CH independently. In another implementation, Q is N, while V is N. 1 V 2 and V 3 Each is N. In another implementation, Q and V... 1 and V 2 It is CH, and V 3 For N. In another implementation, Q, V 2 and V 3 They are CH and V respectively. 1 For N. In another implementation, Q, V 1 and V 3 Both are CH, while V 2 For N. In another implementation, Q and V 2 Both are N, while V 1 and V 3 Both are CH. In another implementation, Q and V 2 Both are CH, while V 1 and V 3 Both are N. In another implementation, Q and V 3 Both are N, while V 1 and V 2 All are CH.

[0433] Unbound by theory, scheme 1a describes the mechanism of release of free drug from camptothecin drug unit, which is linked to a releasable linker as a glucuronide unit via a nitrogen atom from an amine substituent of the free drug.

[0434] Option 1a:

[0435]

[0436] Dispenser (S) * ):

[0437] The camptothecin conjugates described herein may also contain a partitioning agent (S). * The dispensing portion can be used, for example, to mask the hydrophobicity of a specific camptothecin drug unit or other linking unit component.

[0438] Representative dispensing agents include polyethylene glycol (PEG) units, cyclodextrin units, polyamides, hydrophilic peptides, polysaccharides, and dendritic polymers.

[0439] When Q contains polyethylene glycol (PEG) units, cyclodextrin units, polyamides, hydrophilic peptides, polysaccharides, or dendritic polymers, these groups may be present as "in-line" components or as side-chain or branched components. For those embodiments where branched forms are present, the linker unit will typically contain lysine residues (or parallel linker units B), which provide, for example, simple functional coupling between the PEG unit and the remainder of the linker unit.

[0440] Polyethylene glycol (PEG) unit

[0441] Polydisperse PEG, monodisperse PEG, and discrete PEG can be used to prepare the compounds of the present invention. Polydisperse PEG is a non-homogeneous mixture of size and molecular weight, while monodisperse PEG is typically purified from a non-homogeneous mixture and therefore has a single chain length and molecular weight. Preferred PEG units are discrete PEG, which are compounds synthesized stepwise rather than via a polymerization process. Discrete PEG provides a single molecule with a defined and specified chain length.

[0442] The PEG unit described herein comprises one or more polyethylene glycol chains. In some embodiments, the polyethylene glycol chains may be linked together, for example, in a linear, branched, or star configuration. Typically, at least one of the PEG chains is derivatized at one end to covalently attach to a suitable site on a component (e.g., B) of the linker subunit, or may serve as a tandem (e.g., bifunctional) linker group therein to covalently bond two linker subunit components (e.g., ZAS). * -RL-、ZAS * -RL-Y-). The connections within exemplary linker units are either unconditionally cleavable bonds or via conditionally cleavable bonds. Exemplary connections are via amide bonds, ether bonds, ester bonds, hydrazone bonds, oxime bonds, disulfide bonds, peptide bonds, or triazole bonds. In some embodiments, the connections within linker units are unconditionally cleavable bonds. In some embodiments, the connections within linker units are not via ester bonds, hydrazone bonds, oxime bonds, or disulfide bonds. In some embodiments, the connections within linker units are not via hydrazone bonds.

[0443] Conditionally cleavable bonds are those that are essentially insensitive to cleavage when circulating in plasma but sensitive to cleavage in the intracellular or tumor environment. Unconditionally cleavable bonds are those that are essentially insensitive to cleavage in any biological environment. Examples of conditionally cleavable bonds include the chemical hydrolysis of hydrazones, the reduction of disulfides, and the enzymatic cleavage of peptide or glycosidic bonds.

[0444] In some embodiments, the PEG unit is directly connected to the parallel linker unit B. The other end (or terminal) of the PEG unit can be free and unbound and can be in the form of a methoxy, carboxylic acid, alcohol, or other suitable functional group. The methoxy, carboxylic acid, alcohol, or other suitable functional group acts as a cap for the terminal PEG subunit of the PEG unit. Unbound means that the PEG unit is not connected to camptothecin, an antibody, or another linker component at the unbound site. Those skilled in the art will understand that the PEG unit may contain non-PEG material (e.g., to facilitate coupling of multiple PEG chains to each other) in addition to containing repeating polyethylene glycol subunits. Non-PEG material refers to atoms in the PEG unit that are not part of the repeating –CH2CH2O- subunit. In some embodiments provided herein, the PEG unit comprises two monomeric PEG chains linked to each other via a non-PEG component. In other embodiments provided herein, the PEG unit comprises two linear PEG chains connected to a central core or parallel linker unit (i.e., the PEG unit itself is branched).

[0445] Those skilled in the art can utilize numerous PEG-linking methods [see, for example, Goodson, et al. (1990) Bio / Technology 8:343 (PEGylation of interleukin-2 at its glycosylation site after site-directed mutagenesis); EP 0 401 384 (coupling PEG to G-CSF); Malik, et al. (1992) Exp. Hematol. 20:1028-1035 (PEGylation of GM-CSF using tresyl chloride); PCT Publication No. WO 90 / 12874 (PEGylation of erythropoietin containing a recombinantly introduced cysteine ​​residue using a cysteine-specific mPEG derivative); US Patent No. 5,757,078 (PEGylation of EPO peptides); US Patent No. 5,672,662 (Poly(ethyleneglycol) and related polymers monosubstituted with propionic orbutanoic acids and functional derivatives)]. (the thereof for biotechnical applications); US Patent No. 6,077,939 (PEGylation of an N-terminal alpha-carbon of a peptide); Veronese et al., (1985) Appl. Biochem. Bioechnol 11:141-142 (PEGylation of an N-terminal alpha-carbon of a peptide with PEG-nitrophenylcarbonate ("PEG-NPC") or PEG-trichlorophenylcarbonate); and Veronese (2001) Biomaterials 22:405-417 (a review article on peptide and protein PEGylation).

[0446] For example, PEG can be covalently bound to amino acid residues via reactive groups. Reactive groups are those that the activated PEG molecule can bind to (e.g., free amino or carboxyl groups). For example, N-terminal amino acid residues and lysine (K) residues have free amino groups; while C-terminal amino acid residues have free carboxyl groups. Thiol groups (e.g., those present on cysteine ​​residues) can also be used as reactive groups for linking PEG. In addition, enzyme-assisted methods for specifically introducing activating groups (e.g., hydrazides, aldehydes, and aromatic amino groups) at the C-terminus of peptides have been described (see Schwarz et al. (1990) Methods Enzymol. 184:160; Rose et al. (1991) Bioconjugate Chem. 2:154; and Gaertner et al. (1994) J. Biol. Chem. 269:7224).

[0447] In some embodiments, methoxylated PEG ("mPEG") with different reactive moieties can be used to link the PEG molecule to the amino group. Non-limiting examples of such reactive moieties include succinimide succinate (SS), succinimide carbonate (SC), mPEG-imino ester, p-nitrophenyl carbonate (NPC), succinimide propionate (SPA), and cyanuric chloride. Non-limiting examples of such mPEGs include mPEG-succinimide succinate (mPEG-SS), mPEG2-succinimide succinate (mPEG2-SS); mPEG-succinimide carbonate (mPEG-SC), mPEG2-succinimide carbonate (mPEG2-SC); mPEG-imino ester, mPEG-p-nitrophenyl carbonate (mPEG-NPC), mPEG-imino ester; mPEG2-p-nitrophenyl carbonate... PET ester (mPEG2-NPC); mPEG-succinimide propionate (mPEG2-SPA); mPEG2-succinimide propionate (mPEG,--SPA); mPEG-N-hydroxy-succinimide (mPEG-NHS); mPEG2-N-hydroxy-succinimide (mPEG2--NHS); mPEG-cyanuric chloride; mPEG2-cyanuric chloride; mPEG2-lysine alcohol-NPC and mPEG2-Lys-NHS.

[0448] Typically, at least one of the PEG chains constituting a PEG unit is functionalized, enabling it to be covalently linked to other linker components.

[0449] Functionalization includes, for example, via amines, thiols, NHS esters, maleimides, alkynes, azides, carbonyl groups, or some other functional groups. In some embodiments, the PEG unit also comprises a non-PEG material (i.e., a material not composed of –CH2CH2O-) that provides coupling with other linker components or promotes coupling of two or more PEG chains.

[0450] The presence of a PEG unit (or other partitioning agent) in the linker unit may have two potential effects on the pharmacokinetics of the resulting camptothecin conjugate. The desired effect is a decrease in clearance (and consequently an increase in exposure), resulting from reduced nonspecific interactions caused by exposure to the hydrophobic components of the camptothecin conjugate or to the hydrophobic components of camptothecin itself. The second, undesirable effect is a decrease in volume and a reduction in partition rate, which sometimes results from an increase in the molecular weight of the camptothecin conjugate.

[0451] Increasing the number of PEG subunits increases the hydrodynamic radius of the conjugate, which typically leads to reduced diffusivity. This reduced diffusivity, in turn, generally weakens the ability of the camptothecin conjugate to penetrate into the tumor (Schmidt and Wittrup, Mol Cancer Ther 2009; 8:2861-2871). Due to these two competing pharmacokinetic effects, it is desirable to use a sufficiently large PEG to reduce the clearance of the camptothecin conjugate, thereby increasing plasma exposure, but not so large as to significantly reduce its diffusivity to the extent that it interferes with the camptothecin conjugate's ability to reach the intended target cell population. For methods of selecting the optimal PEG size for a specific drug-linker, see examples (e.g., Examples 1, 18, and 21 of US2016 / 0310612, which are incorporated herein by reference).

[0452] In one set of embodiments, the PEG unit comprises one or more linear PEG chains, each chain having at least 2 subunits, at least 3 subunits, at least 4 subunits, at least 5 subunits, at least 6 subunits, at least 7 subunits, at least 8 subunits, at least 9 subunits, at least 10 subunits, at least 11 subunits, at least 12 subunits, at least 13 subunits, at least 14 subunits, at least 15 subunits, at least 16 subunits, at least 17 subunits, at least 18 subunits, at least 19 subunits, at least 20 subunits, at least 21 subunits, at least 22 subunits, at least 23 subunits, or at least 24 subunits. In a preferred embodiment, the PEG unit comprises a total of at least 4 subunits, at least 6 subunits, at least 8 subunits, at least 10 subunits, or at least 12 subunits. In some such embodiments, the PEG unit comprises no more than about 72 subunits in total, preferably no more than about 36 subunits in total.

[0453] In another set of embodiments, the PEG unit comprises a total of 4 to 72, 4 to 60, 4 to 48, 4 to 36, or 4 to 24 subunits; 5 to 72, 5 to 60, 5 to 48, 5 to 36, or 5 to 24 subunits; 6 to 72, 6 to 60, 6 to 48, 6 to 36, or 6 to 24 subunits; 7 to 72, 7 to 60, 7 to 48, 7 to 36, or 7 to 24 subunits; 8 to 72, 8 to 60, 8 to 48, 8 to 36, or 8 to 24 subunits; 9 to 72, 9 to 60, 9 ... 48, 9 to 36 or 9 to 24 subunits; 10 to 72, 10 to 60, 10 to 48, 10 to 36 or 10 to 24 subunits; 11 to 72, 11 to 60, 11 to 48, 11 to 36 or 11 to 24 subunits; 12 to 72, 12 to 60, 12 to 48, 12 to 36 or 12 to 24 subunits; 13 to 72, 13 to 60, 13 to 48, 13 to 36 or 13 to 24 subunits; 14 to 72, 14 to 60, 14 to 48, 14 to 3 6 or 14 to 24 subunits, 15 to 72, 15 to 60, 15 to 48, 15 to 36 or 15 to 24 subunits, 16 to 72, 16 to 60, 16 to 48, 16 to 36 or 16 to 24 subunits, 17 to 72, 17 to 60, 17 to 48, 17 to 36 or 17 to 24 subunits, 18 to 72, 18 to 60, 18 to 48, 18 to 36 or 18 to 24 subunits, 19 to 72, 19 to 60, 19 to 48, 19 to 36 or 19 to 24 subunits, 20 to 72, 20 to 60, 20 to 48, 20 to 36 or 20 to 24 subunits, 21 to 72, 21 to 60, 21 to 48, 21 to 36 or 21 to 24 subunits, 22 to 72, 22 to 60, 22 to 48, 22 to 36 or 22 to 24 subunits, 23 to 72, 23 to 60, 23 to 48, 23 to 36 or 23 to 24 subunits, or 24 to 72, 24 to 60, 24 to 48, 24 to 36 or 24 subunits.

[0454] The following are illustrative linear PEG units that can be used in any of the embodiments provided herein:

[0455]

[0456] The wavy line indicates the connection point with the parallel connector unit (B), and each n is independently selected from 4 to 72, 6 to 72, 8 to 72, 10 to 72, 12 to 72, 6 to 24, or 8 to 24. In some embodiments, the subscript b is about 4, about 8, about 12, or about 24.

[0457] As described herein, the PEG unit is selected to improve the clearance of the resulting camptothecin conjugate without significantly affecting the conjugate's ability to penetrate into the tumor. In embodiments, the PEG unit to be used preferably has 4 to about 24 subunits, more preferably about 4 to about 12 subunits.

[0458] In preferred embodiments of this disclosure, the PEG unit has a density of about 300 Daltons to about 5,000 Daltons; about 300 Daltons to about 4,000 Daltons; about 300 Daltons to about 3,000 Daltons; about 300 Daltons to about 2,000 Daltons; or about 300 Daltons to about 1,000 Daltons. In some such aspects, the PEG unit has at least 6 subunits or at least 8, 10, or 12 subunits. In some such aspects, the PEG unit has at least 6 subunits or at least 8, 10, or 12 subunits, but not more than 72 subunits, preferably not more than 36 subunits.

[0459] It should be understood that when referring to PEG subunits, and depending on the context, the number of subunits may represent an average number, such as when referring to a group of camptothecin conjugates or camptothecin-linker compounds and when using polydisperse PEG.

[0460] Parallel connector unit (B):

[0461] In some embodiments, the camptothecin conjugate and camptothecin linker compound will contain parallel linker units to provide connection sites with the dispensing agent (indicated as -B(S*)- in the linker unit). As a general embodiment, the PEG unit may be linked to a parallel linker unit such as lysine, as shown below, where the wavy line and asterisk indicate covalent bonding within the linker unit of the camptothecin conjugate or camptothecin linker compound:

[0462]

[0463] Spacing sub-unit (Y):

[0464] In some embodiments, the camptothecin conjugates provided herein will have a spacer (Y) between the releasable linker (RL) and camptothecin. The spacer unit may be a functional group that facilitates the linking of RL and camptothecin, or it may provide additional structural components to further facilitate the release of camptothecin from the remainder of the conjugate (e.g., a methylene carbamate unit).

[0465] In embodiments that further promote the release of camptothecin units as free drugs, exemplary spacer subunits are represented by the following formula:

[0466]

[0467] Where EWG is an electron-withdrawing group, R1 It is –H or C1-C4 alkyl, and the subscript n is 1 or 2. In some embodiments, EWG is selected from -CN, -NO2, -CX3, -X, C(=O)OR', -C(=O)N(R')2, -C(=O)R', -C(=O)X, -S(=O)2R', -S(=O)2OR', -S(=O)2NHR', -S(=O)2N(R')2, -P(=O)(OR')2, -P(=O)(CH3)NHR', -NO, -N(R')3 + , wherein X is -F, -Br, -Cl or -I, and R' is independently selected from hydrogen and C1-C6 alkyl, and wherein the wavy line adjacent to the nitrogen atom in each of formulas (a), (a'), (a”), (b) and (b') is a point covalently connected to RL, and the wavy line adjacent to the carbonyl carbon atom in (b) and (b') is a point covalently connected to the heteroatom of the hydroxyl or primary or secondary amine of any of the camptothecin compounds of formulas CPT1, CPT2, CPT3, CPT4, CPT5, CPT6 or CPT7 or any of the compounds 14a-14z in Table I and any of the compounds 18a-18r in Table J, and wherein:

[0468] Formulas (a), (a'), and (a”) represent exemplary methylene carbamate units, wherein T* is a heteroatom of a hydroxyl, primary, or secondary amine functional group from any of the camptothecin compounds of formulas CPT1, CPT2, CPT3, CPT4, CPT5, CPT6, or CPT7, or any of compounds 14a-14z in Table I and any of compounds 18a-18r in Table J, and wherein the wavy line adjacent to T* is a point covalently connected to the remainder of the camptothecin pharmaceutical unit corresponding to the structure of the camptothecin compound.

[0469] In other embodiments, the spacer subunits that serve as methylene carbamate units are represented by the following formula:

[0470]

[0471] Each R is independently H or a C1-C4 alkyl group. Formulas (a1) and (a1') represent methylene carbamate units, wherein O* is a hydroxyl substituent of the lactone ring of a camptothecin compound of formula CPT1, CPT2, CPT3, CPT4, CPT5, CPT6, or CPT7, or any of compounds 14a-14z in Table I and any of compounds 18a-18r in Table J, or another hydroxyl substituent of a camptothecin compound of formula CPT5 or CPT7, or R from CPT6. F Or R F’ The oxygen atom of the hydroxyl substituent, wherein R F and R F’At least one of them is a C1-C8 hydroxyalkyl N,N-(C1-C4 hydroxyalkyl)(C1-C4 alkyl)-amino-C1-C8 alkyl- or N-C1-C4 hydroxyalkyl-C1-C8 aminoalkyl-, C1-C8 alkyl C(O)-,

[0472] Furthermore, the wavy lines in equations (a1), (a1'), and (b1) retain the meanings derived from equations (a), (a'), and (b), respectively. In equation (a1'), –CH2CH2N + (R)2 represents an exemplary basic unit in protonated form.

[0473] Unbound by theory, Scheme 1b describes a mechanism for the release of free drug from camptothecin from a methylene carbamate unit attached to a camptothecin conjugate having a self-eliminating moiety. In this scheme, T* is a heteroatom from a hydroxyl group or a primary or secondary amine of the camptothecin compound incorporated into the methylene carbamate unit.

[0474] Option 1b:

[0475]

[0476] subscript "p"

[0477] In one set of embodiments of the invention, the subscript p denotes the number of drug linker portions on the ligand unit of a single camptothecin conjugate and is preferably an integer in the range of 1 to 16, 1 to 12, 1 to 10, or 1 to 8. A single camptothecin conjugate may also be referred to as a camptothecin conjugate compound. In that set of embodiments, there are 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, or 16 drug linker portions conjugated to a single camptothecin conjugate ligand unit. In another set of embodiments of the invention, camptothecin conjugates describe a group (i.e., camptothecin conjugate compositions) of single camptothecin conjugate compounds that are substantially the same except for the number of camptothecin drug linker portions bonded to each ligand unit, such that the subscript p denotes the average number of camptothecin drug linker portions bonded to the ligand unit of the camptothecin conjugate composition. In this set of embodiments, the subscript p is a number ranging from 1 to about 16, 1 to about 12, 1 to about 10, or 1 to about 8, 2 to about 16, 2 to about 12, 2 to about 10, or 2 to about 8. In some embodiments, the value of the subscript p refers to the average drug load and the drug load of the dominant ADC in the composition.

[0478] In some embodiments, the conjugation will proceed via an interchain disulfide and one to eight camptothecin linker compound molecules will be conjugated with the target agent that becomes the ligand unit. In some embodiments, the conjugation will proceed via an introduced cysteine ​​residue and an interchain disulfide and one to ten, one to twelve, one to fourteen, or one to sixteen camptothecin linker compound moieties will be conjugated with the ligand unit. In some embodiments, the conjugation will proceed via an introduced cysteine ​​residue and two or four camptothecin linker compound molecules will be conjugated with the ligand unit.

[0479] Camptothecin drugs - linker compounds

[0480]

[0481]

[0482] glucuronic acid residues are replaced by mannose.

[0483] mDPR = Maleimide-Aminopropionyl:

[0484] mPR = Maleimide-propionyl:

[0485] PropargOPr=-(C=O)CH2CH2OCH2C≡CH

[0486] Other camptothecin drug linker compounds

[0487]

[0488]

[0489]

[0490] Camptothecin conjugate mixtures and compositions

[0491] This invention provides mixtures of camptothecin conjugates and pharmaceutical compositions comprising any of the camptothecin conjugates described herein. The mixtures and pharmaceutical compositions comprise multiple conjugates. In some embodiments, each conjugate in the mixture or composition is identical or substantially identical; however, the distribution of drug-linker compounds on the ligands in the mixture or composition may vary, and the drug loading may also vary. For example, in some embodiments, the conjugation techniques used to conjugate the drug-linker to the antibody as a target agent may produce compositions or mixtures that are non-uniform in terms of the distribution of camptothecin linker compounds on the antibody (ligand unit) within the mixture and / or composition. In some of those embodiments, in such mixtures or compositions of molecules, the loading of camptothecin linker compounds on each antibody molecule is an integer ranging from 1 to 16.

[0492] In these embodiments, when the composition is referred to as a whole, the drug-linker load is a number ranging from 1 to about 16. A small percentage of unconjugated antibodies may sometimes be present within the composition or mixture. The average number of drug-linkers per ligand unit in the mixture or composition (i.e., the average drug load) is an important property because it relates to the maximum amount of drug that can be delivered to target cells. Typically, the average drug load is 1, 2 or about 2, 3 or about 3, 4 or about 4, 5 or about 5, 6 or about 6, 7 or about 7, 8 or about 8, 9 or about 9, 10 or about 10, 11 or about 11, 12 or about 12, 13 or about 13, 14 or about 14, 15 or about 15, 16 or about 16.

[0493] In some embodiments, the mixture and pharmaceutical composition comprise multiple (i.e., a group) conjugates; however, these conjugates are identical or substantially identical and substantially homogeneous in terms of the distribution of drug-linkers on ligand molecules within the mixture and / or composition and in terms of the loading of drug-linkers on ligand molecules within the mixture and / or composition. In some such embodiments, the loading of drug-linkers on antibody ligand units is 2 or 4. A small percentage of unconjugated antibodies may also be present within the composition or mixture. In such embodiments, the average drug loading is about 2 or about 4. Typically, such compositions and mixtures are produced by the use of site-specific conjugation techniques, and the conjugation is attributed to the introduction of cysteine ​​residues.

[0494] The average number of camptothecin or camptothecin-linker compounds per ligand unit in formulations derived from conjugation reactions is typically characterized using conventional methods such as mass spectrometry, ELISA, and HPLC (e.g., HIC). In those cases, the quantitative distribution of the camptothecin conjugate with respect to subscript p is usually determined. In other cases, the separation, purification, and characterization of homogeneous camptothecin conjugates are typically achieved using conventional methods such as reversed-phase HPLC or electrophoresis.

[0495] In some embodiments, the composition is a pharmaceutical composition comprising the camptothecin conjugate described herein and a pharmaceutically acceptable carrier. In some of those embodiments, the pharmaceutical composition is in liquid form. In other embodiments, the pharmaceutical composition is a lyophilized powder.

[0496] Compositions, including pharmaceutical compositions, may be provided in a purified form. As used herein, "purified" means, when isolated, that the isolate contains at least 95% by weight of the isolate, or at least 98% in other embodiments, a conjugate.

[0497] How to use

[0498] Cancer treatment

[0499] Camptothecin conjugates can be used to inhibit the proliferation of tumor cells or cancer cells, induce apoptosis in tumor or cancer cells, or treat cancer in patients. Camptothecin conjugates are accordingly used in a variety of settings for cancer treatment. Camptothecin conjugates are intended to deliver drugs to tumor cells or cancer cells. Not bound by theory, in one embodiment, the ligand unit of the camptothecin conjugate binds to or conjugates an antigen associated with cancer cells or tumor cells, and the camptothecin conjugate is absorbed (internalized) within the tumor cells or cancer cells via receptor-mediated endocytosis or other internalization mechanisms. In some embodiments, the antigen is linked to tumor cells or cancer cells, or may be an extracellular matrix protein associated with tumor cells or cancer cells. Once inside the cell, the drug is released intracellularly via activation of the activating unit. In an alternative embodiment, the free drug is released from the camptothecin conjugate outside the tumor cells or cancer cells, and the free drug subsequently penetrates the cell.

[0500] In one implementation, the ligand unit binds to tumor cells or cancer cells.

[0501] In another implementation, the ligand unit binds to tumor cell or cancer cell antigens on the surface of tumor cells or cancer cells.

[0502] In another embodiment, the ligand unit binds to a tumor cell or cancer cell antigen, which is an extracellular matrix protein associated with tumor cells or cancer cells.

[0503] The specificity of the ligand unit to specific tumor cells or cancer cells is an important consideration in determining the tumor or cancer to be treated most effectively. For example, camptothecin conjugates targeting cancer cell antigens present in hematopoietic system cancers are used to treat hematologic malignancies (e.g., anti-CD30, anti-CD70, anti-CD19, and anti-CD33 binding ligand units (e.g., antibodies) are used to treat hematologic malignancies). In some embodiments, camptothecin conjugates targeting cancer cell antigens present on solid tumors can be used to treat such solid tumors.

[0504] Cancers intended for treatment with camptothecin conjugates include, but are not limited to, hematopoietic system cancers such as lymphomas (Hodgkin's lymphoma and non-Hodgkin's lymphoma), as well as leukemia and solid tumors. Examples of hematopoietic system cancers include follicular lymphoma, anaplastic large cell lymphoma, mantle cell lymphoma, acute myeloid leukemia, chronic myeloid leukemia, chronic lymphocytic leukemia, diffuse large B-cell lymphoma, and multiple myeloma. Examples of solid tumors include fibrosarcoma, myxosarcoma, liposarcoma, chondrosarcoma, osteosarcoma, chordoma, angiosarcoma, endothelial sarcoma, lymphangiosarcoma, lymphangioendothelial sarcoma, synovoma, mesothelioma, Ewing's tumor, leiomyosarcoma, rhabdomyosarcoma, colon cancer, colorectal cancer, kidney cancer, pancreatic cancer, bone cancer, breast cancer, ovarian cancer, prostate cancer, esophageal cancer, gastric cancer, oral cancer, nasal cancer, pharyngeal cancer, squamous cell carcinoma, basal cell carcinoma, adenocarcinoma, sweat gland carcinoma, sebaceous gland carcinoma, and papillary carcinoma. Cancer, papillary adenocarcinoma, cystic adenocarcinoma, medullary carcinoma, bronchial carcinoma, renal cell carcinoma, liver cancer, bile duct carcinoma, choriocarcinoma, seminoma, embryonal carcinoma, Wilms' tumor, cervical cancer, uterine cancer, testicular cancer, small cell lung cancer, bladder cancer, lung cancer, epithelial carcinoma, glioma, glioblastoma multiforme, astrocytoma, medulloblastoma, craniopharyngioma, ependymoma, pineal tumor, hemangioblastoma, acoustic neuroma, oligodendroglioma, meningioma, neuroblastoma, and retinoblastoma.

[0505] In a preferred embodiment, the cancer being treated is either lymphoma or leukemia.

[0506] Multimodal therapy for cancer

[0507] The intention is to treat or inhibit cancer, including but not limited to tumors, metastases or other diseases or conditions characterized by uncontrolled cell growth, by applying a therapeutically effective amount of camptothecin conjugates.

[0508] In one set of embodiments, a method for treating cancer is provided, comprising administering an effective amount of a camptothecin conjugate and a chemotherapeutic agent to a patient in need. In one embodiment, the chemotherapeutic agent is one for which the cancer has not been found to be refractory. In another embodiment, the chemotherapeutic agent is one for which the cancer has been found to be refractory.

[0509] In another set of embodiments, camptothecin conjugates are administered to patients who have also undergone surgical treatment for cancer. In such embodiments, chemotherapy agents are typically administered over a series of treatment cycles, or one or a combination of chemotherapy agents, such as one or more standard care chemotherapy agents.

[0510] In another set of implementations, the patient also receives additional treatment, such as radiotherapy. In one specific implementation, the camptothecin conjugate is administered concurrently with chemotherapy or radiotherapy. In another specific implementation, chemotherapy or radiotherapy is administered before or after the administration of the camptothecin conjugate.

[0511] Additionally, methods of treating cancer with camptothecin conjugates are offered as alternatives to chemotherapy or radiation therapy, where chemotherapy or radiation therapy has been shown or may be shown to be too toxic to the treated subject, for example, causing unacceptable or intolerable side effects. The treated patient may optionally be treated with another cancer treatment such as surgery, radiation therapy, or chemotherapy, depending on which treatment is found to be acceptable or tolerable.

[0512] Treatment of autoimmune diseases

[0513] Camptothecin conjugates are intended to kill or inhibit unwanted replication of cells that produce autoimmune diseases or to treat autoimmune diseases.

[0514] Camptothecin conjugates are accordingly used in a variety of settings to treat autoimmune diseases in patients. Camptothecin conjugates are commonly used to deliver camptothecin drugs to target cells. Unbound by theory, in one embodiment, the camptothecin conjugate conjugate is conjugated to an antigen on the surface of pro-inflammatory or inappropriately stimulated immune cells, and then absorbed into the target cells via receptor-mediated endocytosis. Once inside the cell, the linker unit is cleaved, resulting in the release of the camptothecin drug unit as a free drug. The free camptothecin drug can then migrate within the cytosol and induce cytotoxic or inhibitory cell activity. In an alternative embodiment, the camptothecin drug unit is cleaved from the camptothecin conjugate outside the target cells, subsequently releasing the resulting free camptothecin drug that permeates the cells.

[0515] In one implementation, the ligand unit binds to an autoimmune antigen. In this implementation, the antigen is on the surface of the cells involved in the autoimmune disease.

[0516] In one implementation, the ligand unit binds to activated lymphocytes associated with an autoimmune disease state.

[0517] In yet another implementation, camptothecin conjugates kill or inhibit the proliferation of cells that produce autoantibodies associated with specific autoimmune diseases.

[0518] Specific types of autoimmune diseases for which camptothecin conjugates are intended for treatment include, but are not limited to, Th2 lymphocyte-related diseases (e.g., atopic dermatitis, atopic asthma, rhinoceros conjunctivitis, allergic rhinitis, Omenn syndrome, systemic sclerosis, and graft-versus-host disease); Th1 lymphocyte-related diseases (e.g., rheumatoid arthritis, multiple sclerosis, psoriasis, Sjögren's syndrome, Hashimoto's thyroiditis, Grave's disease, primary biliary cirrhosis, Wegener's granulomatosis, and tuberculosis); and activated B lymphocyte-related diseases (e.g., systemic lupus erythematosus, pulmonary hemorrhage-nephritis syndrome, rheumatoid arthritis, and type I diabetes).

[0519] Multidrug therapy for autoimmune diseases

[0520] The method for treating autoimmune diseases is also disclosed, which involves administering an effective amount of camptothecin conjugate and another therapeutic agent known for treating autoimmune diseases to a patient in need.

[0521] Composition and method of application

[0522] This invention provides pharmaceutical compositions comprising the camptothecin conjugate described herein and at least one pharmaceutically acceptable carrier. The pharmaceutical composition is in any form that allows the compound to be administered to a patient to treat a condition related to the expression of an antigen bound to the ligand unit. For example, the conjugate is in liquid or solid form. A preferred route of administration is parenteral. Parenteral administration includes subcutaneous injection, intravenous, intramuscular, intrasternal injection, or infusion techniques. In one embodiment, the composition is administered parenterally. In one embodiment, the conjugate is administered intravenously. Administration is carried out by any convenient route, such as by infusion or bolus.

[0523] Pharmaceutical compositions are formulated such that the camptothecin conjugate is bioavailable after administration to a patient. The compositions are sometimes in the form of one or more dosage units.

[0524] The materials used in the preparation of the pharmaceutical composition are preferably non-toxic in the amount used. It will be apparent to those skilled in the art that the optimal dosage of one or more active ingredients in a pharmaceutical composition will depend on a variety of factors. These factors include, but are not limited to, the type of animal (e.g., human), the specific form of the compound, the method of administration, and the composition used.

[0525] In some embodiments, the composition is in liquid form. In some of those embodiments, the liquid is used for delivery by injection. In some embodiments, in addition to camptothecin conjugates, the composition for injection administration contains one or more excipients selected from: surfactants, preservatives, wetting agents, dispersants, suspending agents, buffers, stabilizers, and isotonic agents.

[0526] In some embodiments, the liquid compositions, whether solutions, suspensions, or other similar forms, comprise one or more of the following: a sterile diluent (such as water for injection), an aqueous saline solution (preferably physiological saline), Ringer's solution, isotonic sodium chloride, a non-volatile oil that can act as a solvent or suspension medium, such as synthetic mono- or diglycerides of glycerol, polyethylene glycol, glycerol, cyclodextrin, propylene glycol, or other solvents; an antibacterial agent, such as benzyl alcohol or methylparaben; an antioxidant, such as ascorbic acid or sodium bisulfite; a chelating agent, such as ethylenediaminetetraacetic acid; a buffer, such as amino acids, acetates, citrates, or phosphates; a detergent, such as a nonionic surfactant or a polyol; and a tension modulator, such as sodium chloride or dextrose. Parenteral compositions are sometimes packaged in ampoules, disposable syringes, or multi-dose vials made of glass, plastic, or other materials. Physiological saline is an exemplary adjuvant. Injectable compositions are preferably sterile.

[0527] In some implementations, the amount of conjugate effective in treating a particular condition or symptom will depend on the nature of the disease or symptom and will be determined using standard clinical techniques. Additionally, in vitro or in vivo assays may be optionally employed to help identify the optimal dosage range. The precise dosage to be used in the composition also depends on the route of administration and the severity of the disease or symptom, and should be determined based on the practitioner's judgment and the individual patient's situation.

[0528] The composition contains an effective amount of camptothecin conjugate to obtain a suitable dosage. Typically, this amount is at least about 0.01% by weight of the compound in the composition.

[0529] For intravenous administration, the pharmaceutical composition typically comprises about 0.01 to about 100 mg of camptothecin conjugate per kg of animal body weight. In one embodiment, the composition may comprise about 1 to about 100 mg of camptothecin conjugate per kg of animal body weight. In another aspect, the amount administered will be in the range of about 0.1 to about 25 mg of compound / kg body weight. Depending on the drug used, the dose may be even lower, for example, from 1.0 μg / kg to 5.0 mg / kg, 4.0 mg / kg, 3.0 mg / kg, 2.0 mg / kg, or 1.0 mg / kg, or from 1.0 μg / kg to 500.0 μg / kg of subject body weight.

[0530] Typically, the dose of the conjugate administered to the patient is typically from about 0.01 mg / kg to about 100 mg / kg of the subject's body weight, or from 1.0 μg / kg to 5.0 mg / kg of the subject's body weight. In some embodiments, the dose administered to the patient is between about 0.01 mg / kg and about 15 mg / kg of the subject's body weight. In some embodiments, the dose administered to the patient is between about 0.1 mg / kg and about 15 mg / kg of the subject's body weight. In some embodiments, the dose administered to the patient is between about 0.1 mg / kg and about 20 mg / kg of the subject's body weight. In some embodiments, the dose administered is between about 0.1 mg / kg and about 5 mg / kg, or between about 0.1 mg / kg and about 10 mg / kg of the subject's body weight. In some embodiments, the dose administered is between about 1 mg / kg and about 15 mg / kg of the subject's body weight. In some embodiments, the dose administered is between about 1 mg / kg and about 10 mg / kg of the subject's body weight. In some implementations, the dose administered during a treatment cycle is between about 0.1 to 4 mg / kg, or even more preferably 0.1 to 3.2 mg / kg, or even more preferably 0.1 to 2.7 mg / kg of the subject's body weight.

[0531] The term "carrier" refers to a diluent, adjuvant, or excipient that is administered with a compound. In some embodiments, such drug carriers are liquids, such as water and oils, including those of petroleum, animal, plant, or synthetic origin, such as peanut oil, soybean oil, mineral oil, and sesame oil. Other carriers include saline, gum arabic, gelatin, starch paste, talc, keratin, colloidal silica, and urea. Additionally, excipients, stabilizers, thickeners, lubricants, and colorants are sometimes used. In one embodiment, the camptothecin conjugate and its combinations, along with the pharmaceutically acceptable carrier, are sterile when administered to a patient.

[0532] Water is an exemplary carrier when the compound is administered intravenously. Saline solutions, as well as aqueous solutions of dextran and glycerol, are frequently used as liquid carriers, especially for injectable solutions. Suitable drug carriers also include excipients such as starch, glucose, lactose, sucrose, gelatin, malt, rice, flour, chalk, silica gel, sodium stearate, glyceryl monostearate, talc, sodium chloride, skim milk powder, glycerol, propylene, ethylene glycol, water, and ethanol. If desired, the compositions of the present invention may also contain small amounts of wetting agents, emulsifiers, or pH buffers.

[0533] In one embodiment, the conjugate is formulated into a pharmaceutical composition suitable for intravenous administration to animals, particularly humans, according to standard procedures. Typically, the carrier or medium for intravenous administration is a sterile isotonic buffer solution. If necessary, the composition contains a solubilizer. The composition for intravenous administration optionally contains a local anesthetic such as lidocaine to relieve pain at the injection site. Typically, the components are provided separately or mixed together in unit dosage forms, for example, as a dry lyophilized powder or anhydrous concentrate in a closed container such as an ampoule or capsule indicating the amount of active agent. When the conjugate is administered by infusion, it is typically dispensed, for example, in an infusion bottle containing sterile pharmaceutical-grade water or saline. When the conjugate is administered by injection, sometimes an ampoule of sterile water for injection or saline is provided so that the components can be mixed prior to administration.

[0534] Pharmaceutical compositions are typically formulated to be sterile, substantially isotonic, and fully compliant with all Good Manufacturing Practices (GMP) requirements of the U.S. Food and Drug Administration.

[0535] Methods for preparing camptothecin conjugates

[0536] The camptothecin conjugates described herein are prepared in a tandem configuration of antibody, linker, and drug unit, or by assembling the components and then completing the assembly steps in a convergent manner. The Coursius rearrangement or chloramine synthesis can be used to provide methyl carbamate linkers (spacers), which are used in many embodiments of the conjugates described herein.

[0537] Option 2: Prepare exemplary camptothecin drug-linker compounds of the formula Z'-A-RL-YD, Z'-AS*-RL-YD, or Z'-AB(S*)-RL-YD using the Curvature rearrangement reaction, wherein Y has the formula (a'):

[0538]

[0539] Scheme 2 illustrates a synthetic strategy for the Coursius rearrangement of acyl azide derivatives involving free drug, where CPT is the camptothecin drug unit, the structure of which corresponds to a camptothecin compound with a hydroxyl functional group, the oxygen atom of the hydroxyl functional group represented by O* being incorporated into a methylene carbamate unit formed as a rearrangement, Z' is the elongation unit precursor, RL is the releasable linker, and X is -A-, -AS*-, or –AB(S*)-, where A is the linker unit, S* is the partitioning agent, and B is the parallel linker unit. This strategy is applicable to camptothecin drugs containing multiple alcohols or other heteroatoms as a means of obtaining regioselectivity, as there are many complementary alkylation methods to form acyl azides such as: alkylation of haloesters, alkylation of haloacids, or insertion of metal carbides with ethyl diazonate or methyl diazonate; see Doyle, M. et al., Modern Catalytic Methods for Organic Synthesis with Diazo Compounds; Wiley: New York, 1998. The acyl azide is then heated together with at least a stoichiometric amount of an alcohol-containing linker intermediate of the formula Z'-X-RL-OH.

[0540] Option 3: Alternative preparation of exemplary camptothecin drug-linker compounds of the formula Z'-A-RL-YD, Z'-AS*-RL-YD, or Z'-AB(S*)-RL-YD via N-chloromethylamine, wherein the spacer subunit Y is a methylene carbamate unit of formula (a) or (a'):

[0541]

[0542] Where R 1 It is hydrogen or C1-C4 alkyl, R is -H or –CH2CH2SO2Me, and the meaning of other variable groups comes from Scheme 2.

[0543] The synthesis of N-chloromethylamine is an alternative to the Coursius rearrangement because it allows the introduction of unmodified alcohols or other camptothecin compounds containing heteroatoms, the use of which may be incompatible with the conditions required to form the acyl azide of Scheme 2, and is carried out by condensation with reactive N-chloromethylamine. This method is also more suitable for introducing certain types of methylene carbamate units, as shown, for example, in Scheme 4.

[0544] Scheme 4 clarifies the synthesis of exemplary camptothecin-linker compounds in which the spacer subunit (Y) is a methylene carbamate unit of formula (a”) of formula Z'-A-RL-YD, Z'-AS*-RL-YD, or Z'-AB(S*)-RL-YD. The reaction of nitrophenyl carbonate with a cyclic amino group provides a carbamate, which is then converted to a chlorocycloalkylamine for alkylation with a nucleophilic reagent from the mercapto, hydroxyl, amine, or amide functional groups of the free camptothecin drug. Alternatively, the drug moiety can be stored... The carbamate was treated with acid to assemble the drug-linker intermediate shown. The alkylation product was deprotected, and the resulting free amine was then condensed with N-hydroxysuccinimide 3-maleimide propionate to introduce an extended subunit precursor covalently linked to the linker unit, thereby providing a camptothecin-linker compound. The resulting camptothecin-linker compound was then condensed with a thiol-containing target agent to provide a camptothecin conjugate having a spacer subunit comprising a self-eliminating moiety and a carbamate methylene ester unit of formula a”.

[0545] Option 4: Prepare an exemplary camptothecin drug linker compound of formula Z'-A-RL-YD, wherein the spacer unit Y is a methylene carbamate unit of formula (a”).

[0546]

[0547] For camptothecin-linker compounds and camptothecin conjugates containing a methylene carbamate unit, where T* is the nitrogen atom of a primary or secondary amine substituent of the camptothecin compound, direct alkylation with chloromethylamine after the general steps provided in Scheme 3 or Scheme 4 may not be suitable because the nitrogen heteroatom of the amine functional group from the free drug is over- or undesirably over-alkylated. In those cases, the method embodied in Scheme 5 can be used.

[0548] Option 5

[0549]

[0550] In Scheme 5, an intermediate carbamate is prepared, which already has a basic unit (i.e., a dimethylaminoethyl moiety) as an R substituent for a carbamate unit of formula (a1'). The nitrogen of this carbamate is condensed with formaldehyde, and the resulting intermediate is quenched with the amine functional group of a camptothecin drug containing an aliphatic amine. N* represents the nitrogen atom from this functional group. This condensation forms a carbamate of formula (a1') covalently linked to the camptothecin drug unit, wherein R... 1 R is hydrogen and is dimethylaminoethyl. The phenyl nitro group is then reduced to an amine, thereby providing a process for the sequential introduction of the linker unit (A) and the extension subunit precursor (Z').

[0551] Implementation plan with numbering

[0552] The following numbered embodiments describe various non-limiting aspects of the invention.

[0553] 1. A method having the formula L-(QD) p Camptothecin conjugates or salts thereof, wherein L is a ligand unit; subscript p is an integer from 1 to 16; Q is a linker unit having the following formulas: -ZA-, -ZA-RL-, -ZA-RL-Y-, -ZAS * -RL-、-ZAS * -RL-Y-、ZAS * -W-、-ZAS * -W-RL-、-ZAB(S * )-RL-、-ZAB(S * )-W-、-ZAB(S * -W-RL- and -ZAB(S * )-RL-Y-, where Z is the extended sub-element; A is the key or connector element; B is the parallel connector element; S * RL is a dispensing agent; W is a releasable linker; Y is a spacer unit; and D is a pharmaceutical unit selected from the following CPT1, CPT2, CPT3, CPT4, CPT5, CPT6, and CPT7:

[0554]

[0555]

[0556] Where R B The derivative is selected from H, C1-C8 alkyl, C1-C8 haloalkyl, C3-C8 cycloalkyl, (C3-C8 cycloalkyl)-C1-C4 alkyl, phenyl, and phenyl-C 1- Members of C4 alkyl groups; R C Selected from C1-C6 alkyl and C 3- Members of C6 cycloalkyl groups; R F and R F’ Each is independently selected from -H, C1-C8 alkyl, C1-C8 hydroxyalkyl, C1-C8 aminoalkyl, (C1-C4 alkylamino)-C1-C8 alkyl, N,N-(C1-C4 hydroxyalkyl)(C1-C4 alkyl)amino-C1-C8 alkyl, N,N-bis(C1-C4 alkyl)amino-C 1- C8 alkyl, N-C1-C4 hydroxyalkyl-C1-C8 aminoalkyl, C1-C8 alkyl C(O)-, C1-C8 hydroxyalkyl-C(O)-, C1-C8 aminoalkyl-C(O)-, C3-C 10 cycloalkyl, (C3-C10 Cycloalkyl)-C1-C4 alkyl, C3-C 10 Heterocyclic alkyl groups, (C3-C 10 Members of heterocyclic alkyl (C1-C4 alkyl), phenyl, phenyl-C1-C4 alkyl, diphenyl-C1-C4 alkyl, heteroaryl and heteroaryl-C1-C4 alkyl; or R F and R F’ It combines with the nitrogen atoms to form a 5-, 6-, or 7-membered ring having 0 to 3 substituents selected from halogens, C1-C4 alkyl groups, -OH, -OC1-C4 alkyl groups, -NH2, and -NHC. 1- C4 alkyl and -N(C1-C4 alkyl)2; and wherein R B R C R F and R F’ The cycloalkyl, heterocycloalkyl, phenyl, and heteroaryl moieties are composed of 0 to 3 elements selected from halogens, C 1- C4 alkyl, -OH, -OC1-C4 alkyl, -NH2, -NHC 1- C4 alkyl and -N(C 1- Substituents of C4 alkyl group 2; or

[0557] D is a pharmaceutical unit selected from 13a-13c of Table H, 14a-14z of Table I and 18a-18r of Table J.

[0558] And where Q is -ZA-RL-, -ZA-RL-Y-, -ZAS * -RL-、-ZAB(S * -RL-、-ZAS * -RL-Y- or -ZAB(S) * When )-RL-Y-, the covalent bonding site of D is any one of the hydroxyl, aliphatic primary amino, or secondary amino substituents in any one of CPT1, CPT2, CPT3, CPT4, CPT5, CPT6, and CPT7, or any one of the heteroatoms in any one of the substituents in 13a-13c of Table H, 14a-14z of Table I, and 18a-18r of Table J.

[0559] Where Q is -ZA- or -ZAS * -W- or -ZAB(S * -W-, or when Q is -ZAS * -RL-、-ZAB(S * -RL-、-ZAS * -W-RL- or -ZAB(S *When RL is a releasable unit other than a glucuronide unit, the covalent linker of D is the oxygen atom of the hydroxyl substituent in the lactone ring of any one of CPT1, CPT2, CPT3, CPT4, CPT5, CPT6, or CPT7, or any one of 13a-13c in Table H, 14a-14z in Table I, and 18a-18r in Table J; and

[0560] The condition is that when the covalent linkage site is the nitrogen atom of an aliphatic primary or secondary amino substituent in CPT6, R F and R F’ At least one of them is -H, and the condition is that when D is a compound CPT1 having nitrogen atoms covalently linked through its amino substituents, -ZA-RL-, -ZA-RL-Y-, -ZAS * -RL-、-ZAB(S * -RL-、-ZAS * -RL-Y- and -ZAB(S * The -ZA- of )-RL-Y- is not a succinimide-hexanoyl-β-alanyl moiety, which optionally has a succinimide ring in a hydrolyzed form as a succinic amide moiety.

[0561] 2. The camptothecin conjugate according to embodiment 1, wherein Q is a linker unit having the formula selected from –ZA-RL-, -ZA-RL-Y-, -ZAS*-RL-, -ZAB(S*)-RL-, -ZAS*-RL-Y- and –ZAB(S*)-RL-Y-, wherein A is a linker unit and RL is a glucuronic acid unit.

[0562] 3. The camptothecin conjugate according to embodiment 1 or 2, wherein the covalent linking point of D is the oxygen atom of the hydroxyl substituent on the lactone ring of any one of CPT1, CPT2, CPT3, CPT4, CPT5, CPT6 and CPT7.

[0563] 4. The camptothecin conjugate according to embodiment 1 or 2, wherein the covalent linking point of D is the oxygen atom of the hydroxyl substituent on the lactone ring of any one of Table H 13a-13c, Table I 14a-14z or Table J 18a-18r.

[0564] 5. Camptothecin conjugate according to implementation scheme 1 or 2, wherein the covalent linkage site of D is through R of CPT6. F Or R F’ The oxygen atom of the hydroxyl substituent, wherein R F and R F’At least one of them is C1-C8 hydroxyalkyl N,N-(C1-C4 hydroxyalkyl)(C1-C4 alkyl)-amino-C1-C8 alkyl-, N-C1-C4 hydroxyalkyl-C1-C8 aminoalkyl-, or C1-C8 alkyl C(O)-.

[0565] 6. Camptothecin conjugate according to implementation scheme 1 or 2, wherein the covalent linkage site of D is through R of CPT6. F Or R F’ The nitrogen atom of the amine substituent, where R F and R F’ At least one of them is C1-C8 aminoalkyl, (C1-C4 alkylamino)-C1-C8 alkyl-, N-(C1-C4 hydroxyalkyl)-C1-C8 aminoalkyl- or C1-C8 aminoalkylC(O)-.

[0566] 7. Camptothecin conjugate according to embodiment 1 or 2, wherein the covalent linking point of D is the nitrogen atom of the amino substituent of CPT1, provided that -ZA- is not succinimide-hexanoyl-β-alanyl group with an optional hydrolyzed succinimide ring as the succinimide moiety.

[0567] 8. The camptothecin conjugate according to embodiment 1 or 2, wherein the covalent linking point of D is the nitrogen atom of the amino substituent of CPT4.

[0568] 9. A camptothecin conjugate according to embodiment 1 or 2, wherein the covalent linkage is through the nitrogen atom of the substituent on CPT6, provided that R F and R F’ At least one of them is -H.

[0569] 10. A camptothecin conjugate according to any one of embodiments 1-9, wherein Q is a connecting subunit having a formula selected from -ZA-RL- and -ZA-RL-Y-.

[0570] 11. A camptothecin conjugate according to any one of embodiments 1-9, wherein Q is a compound having a derivative selected from -ZAS. * -RL- and -ZAS * -RL-Y- style connection subunit.

[0571] 12. A camptothecin conjugate according to any one of embodiments 1-9, wherein Q is a connecting subunit having a formula selected from -ZAB(S*)-RL- and –ZAB(S*)-RL-Y-.

[0572] 13. A camptothecin conjugate according to any one of implementation schemes 1-3 and 7, wherein D is CPT1.

[0573] 14. A camptothecin conjugate according to any one of implementation schemes 1-3, wherein D is CPT2.

[0574] 15. A camptothecin conjugate according to any one of implementation schemes 1-3, wherein D is CPT3.

[0575] 16. A camptothecin conjugate according to any one of implementation schemes 1-3 and 8, wherein D is CPT4.

[0576] 17. A camptothecin conjugate according to any one of implementation schemes 1-3, wherein D is CPT5.

[0577] 18. A camptothecin conjugate according to any one of implementation schemes 1-3, 5, 6 and 9, wherein D is CPT6.

[0578] 18. A camptothecin conjugate according to any one of implementation schemes 1-3, wherein D is CPT7.

[0579] 19. A camptothecin conjugate according to any one of embodiments 2-18, wherein RL is a glucuronic acid unit having the following formula:

[0580]

[0581] Where Su is the hexose form of a monosaccharide, particularly a glucuronic acid or mannose residue; O' represents the oxygen atom of a glycosidic bond that can be cleaved by glycosidases; the wavy line marked with a single asterisk (*) indicates that R is related to the oxygen atom in the glycosidic bond. F and R F’ At least one of the following is a site covalently linked to the nitrogen atom of an aliphatic primary or secondary amino substituent of any of the following CPT1, CPT4, CPT6 of -H, or 13b and 13c of Table H, 14a-14f and 14i-14o, 14s and 14u-14z of Table I, and 18q and 18r of Table J, or a site covalently linked to the spacer subunit (Y), or a site covalently linked to the oxygen atom of a hydroxyl substituent in the lactone ring of any of the following CPT1, CPT2, CPT3, CPT4, CPT5, CPT6 or CPT7, or 13a-13c of Table H, 14a-14z of Table I, or 18a-18r of Table J; and a wavy line marked with a double asterisk (**) indicates a site covalently linked to the remainder of Q.

[0582] 20. The camptothecin conjugate according to embodiment 19, wherein Q is a connecting subunit having the formula -ZA-RL-Y-, -ZAS*-RL-Y-, or -ZAB(S*)-RL-Y-; and the spacer subunit (Y) has the following formula:

[0583]

[0584] Where EWG is an electron-withdrawing group; O* represents the oxygen atom from the hydroxyl functional group of D; the wavy line adjacent to the nitrogen atom indicates the site covalently bonded to the carbonyl carbon atom of the glucuronide unit; and the wavy line adjacent to O* indicates the site covalently bonded to the rest of D.

[0585] 21. The camptothecin conjugate according to embodiment 19, wherein Q is a connecting subunit having the formula -ZA-RL-Y-, -ZAS*-RL-Y-, or -ZAB(S*)-RL-Y-; D is selected from R F and R F’ Each of them is -H CPT1, CPT4 and CPT6, and the spacer subunit (Y) has the following formula:

[0586]

[0587] Where EWG is an electron-withdrawing group; the wavy line adjacent to the nitrogen atom indicates the site covalently bonded to the carbonyl carbon atom of the glucuronide unit; and the wavy line adjacent to the carbonyl carbon atom indicates the site covalently bonded to the R atom. F and R F’ Each of these is a site where the nitrogen atom of an amino substituent of CPT1, CPT4, or CPT6 is covalently linked.

[0588] 22. The camptothecin conjugate according to embodiment 19, wherein Q is a connecting subunit having the formula -ZA-RL-Y-, -ZAS*-RL-Y-, or -ZAB(S*)-RL-Y-; D is selected from R F and R F’ Each of them is -H CPT1, CPT4 and CPT6, and the spacer subunit (Y) has the following formula:

[0589]

[0590] Where EWG is an electron-withdrawing group; the wavy line adjacent to the nitrogen atom indicates the site covalently bonded to the carbonyl carbon atom of the glucuronide unit; and the wavy line adjacent to the carbonyl carbon atom indicates the site covalently bonded to the R atom. F and R F’ Each of these is a site where the nitrogen atom of an amino substituent of CPT1, CPT4, or CPT6 is covalently linked.

[0591] 23. A camptothecin conjugate according to any one of embodiments 1-22, wherein A is a linker unit, wherein the linker unit is composed of a triazole moiety, wherein the triazole moiety is optionally formed by 1,3-dipolar cycloaddition of an azide substituent from a chemically modified target agent, said target agent being a precursor of a conjugate ligand unit of an alkynyl moiety of a drug linker compound.

[0592] 24. A camptothecin conjugate according to any one of embodiments 1-22, wherein -ZA- is composed of a succinimide-alkanoyl moiety or a succinimide and triazole moiety, each optionally having a hydrolyzed succinimide ring as the succinamide moiety, or a succinamide moiety derivatized from the mDPR moiety of the camptothecin-linker compound, provided that -ZA- is composed of a succinimide and triazole moiety, optionally having a hydrolyzed succinimide ring as the succinamide moiety, or a succinamide moiety derivatized from the mDPR moiety when CPT1 is covalently linked through the nitrogen atom of its amino substituent.

[0593] 25. A camptothecin conjugate according to any one of embodiments 1-22, wherein -ZA- is composed of a succinimide-alkanoyl-β-alanyl moiety, which optionally has a succinimide ring in a hydrolyzed form as the succinic amide moiety, provided that when D is CPT1, D has a covalently bonded oxygen atom to a hydroxyl substituent on its lactone ring.

[0594] 26. A camptothecin conjugate according to any one of embodiments 1-22, wherein -ZA- is composed of a succinic amide moiety derived from the mDPR portion of the camptothecin-linker compound.

[0595] 27. A camptothecin conjugate or a salt thereof according to embodiment 26, wherein Q has the following formula:

[0596]

[0597] The succinimide ring is the hydrolyzed form of the succinic amide moiety; the wavy line marked with a single asterisk (*) indicates that R... F and R F’ It is a site covalently linked to the nitrogen atom of CPT1, CPT4 or CPT6 of -H, or a site covalently linked to the spacer subunit; and a wavy line marked with a three-star symbol (***) indicates a site covalently linked to the sulfur atom of L.

[0598] 28. A camptothecin conjugate according to any one of embodiments 2-26, wherein RL is a glucuronic acid unit having the following formula:

[0599]

[0600] The wavy line marked with a single asterisk (*) indicates the oxygen atom of the hydroxyl substituent on the lactone ring of CPT1, CPT2, CPT3, CPT4, CPT5, CPT6, or CPT7, or the oxygen atom of the R substituent therein. F and R F’Each of these is a nitrogen atom of an amino substituent of CPT1, CPT4, or CPT6 with -H, or a point covalently connected to a spacer subunit (Y); and the wavy lines marked with double asterisks (**) indicate connections to A, B, and S. * Or points covalently connected by Z.

[0601] 29. The camptothecin conjugate according to embodiment 28, wherein Q is a linker unit having the formula selected from -ZAS*-RL-; -ZAB(S*)-RL-; -ZAS*-RL-Y-; and -ZAB(S*)-RL-Y-, wherein -ZA- is composed of a succinimide-propionyl moiety.

[0602] 30. The camptothecin conjugate according to embodiment 29, wherein Q is a compound having the following formula: -ZAS*-RL-

[0603]

[0604] The subscript n is an integer from 1 to 50; the wavy line marked with a single asterisk (*) indicates the site of covalent connection with D or spacer subunit (Y); and the wavy line marked with a triplet (***) indicates the point of covalent connection with sulfur atom of L.

[0605] 31. The camptothecin conjugate according to implementation scheme 30, wherein the subscript n is 4.

[0606] 32. The camptothecin conjugate or its salt according to embodiment 2, wherein -QD has the following structure:

[0607]

[0608] It may optionally have a succinimide ring in a hydrolyzed form as the succinamide moiety, where the wavy line indicates a covalent connection with the sulfur atom of the ligand unit.

[0609] 33. The camptothecin conjugate or its salt according to embodiment 2, wherein -QD has the following structure:

[0610]

[0611] The wavy line indicates a covalent bond with the sulfur atom of the ligand unit, and the succinimide ring is the hydrolyzed form of the succinamide moiety.

[0612] 34. The camptothecin conjugate or its salt according to embodiment 2, wherein -QD has the following structure:

[0613]

[0614] It may optionally have a succinimide ring in a hydrolyzed form as the succinamide moiety, where the wavy line indicates a covalent connection with the sulfur atom of the ligand unit.

[0615] 35. A camptothecin conjugate according to any one of embodiments 1-34, wherein L is derived from an antibody.

[0616] 36. The camptothecin conjugate according to embodiment 35, wherein the antibody specifically binds to an antigen selected from CD19, CD30, CD33, CD70 and LIV-1.

[0617] 37. The camptothecin conjugate according to embodiment 1, wherein Q is a linker unit having a formula selected from -ZA-, -ZAS*-W- and -ZAB(S*)-W-, wherein A is a linker unit, or Q is a linker unit having a formula selected from -ZA-RL-, -ZAS*-RL-, -ZAB(S*)-RL-, -ZAS*-W-RL- and –ZAB(S*)-W-RL, wherein A is a linker unit and RL is a releasable linker other than a glucuronide unit.

[0618] 38. The camptothecin conjugate according to embodiment 37, wherein Q is a connecting subunit having the formula -ZAS*-W- or -ZAS*-W-RL-.

[0619] 39. The camptothecin conjugate according to embodiment 37, wherein Q is a compound having the formula -ZA-RL- or -ZAS. * -RL- Connector Subunit.

[0620] 40. The camptothecin conjugate according to embodiment 37, wherein Q is a connecting subunit having the formula -ZA-.

[0621] 41. A camptothecin conjugate according to any one of implementation schemes 37-40, wherein D is CPT2.

[0622] 42. A camptothecin conjugate according to any one of implementation schemes 37-40, wherein D is CPT3.

[0623] 43. A camptothecin conjugate according to any one of implementation schemes 37-40, wherein D is CPT1.

[0624] 44. A camptothecin conjugate according to any one of implementation schemes 37-40, wherein D is CPT4.

[0625] 45. A camptothecin conjugate according to any one of implementation schemes 37-40, wherein D is CPT5.

[0626] 46. ​​A camptothecin conjugate according to any one of implementation schemes 37-40, wherein D is CPT6.

[0627] 47. A camptothecin conjugate according to any one of implementation schemes 37-40, wherein D is CPT7.

[0628] 48. A camptothecin conjugate according to any one of embodiments 37-47, wherein Q is a connecting subunit having the formula selected from –ZA-RL-, -ZAS*-RL- and –ZAS*-W-RL-, and RL has the following formula:

[0629]

[0630] The wavy lines marked with double asterisks (**) indicate sites covalently connected to D; and the wavy lines marked with single asterisks (*) indicate sites covalently connected to A, S*, or W.

[0631] 49. The camptothecin conjugate according to embodiment 48, wherein -QD has the formula -ZAS*-W-RL-D, where D is the formula where R is the camptothecin conjugate. F and R F’ Each of them is a -H CPT1, CPT4, or CPT6, which is covalently linked to the nitrogen atom of the amine functional group; and W is an amino acid unit selected from N-methyl-glycine (sarcosine), N-methyl-alanine, N-methyl-β-alanine, valine, N-methyl-valine, or D is a CPT1, CPT2, CPT3, CPT4, CPT5, CPT6, or CPT7 covalently linked to the oxygen atom of the hydroxyl substituent on the lactone ring; and W is an amino acid unit selected from glutamic acid or lysine.

[0632] 50. A camptothecin conjugate according to any one of embodiments 37-49, wherein -ZA- is composed of a succinimide-alkanoyl moiety or a succinimide and triazole moiety, each optionally having a succinimide ring in a hydrolyzed form as the succinamide moiety, or a succinamide moiety derived from the mDPR of the camptothecin-linker compound.

[0633] 51. A camptothecin conjugate according to any one of embodiments 37-49, wherein -ZA- is composed of a succinimide-alkanoyl moiety having optionally been hydrolyzed as a succinimide ring as a succinic amide moiety.

[0634] 52. A camptothecin conjugate according to any one of embodiments 37-49, wherein -ZA- is composed of a succinic acid amide moiety derived from the mDPR portion of the camptothecin-linker compound.

[0635] 53. A camptothecin conjugate according to any one of embodiments 37-49, wherein Q is a connecting subunit having a formula selected from -ZAS*-RL- and –ZAS*-W-RL-, wherein -ZA- has a formula selected from:

[0636]

[0637] The succinimide ring in hydrolyzed form is selected as the succinamide moiety, wherein the wavy line marked with a double asterisk (**) indicates the site covalently linked to S*; and the wavy line marked with a triple star (***) indicates the point covalently linked to the sulfur atom of L.

[0638] 54. A camptothecin conjugate according to any one of embodiments 37-49, wherein Q is a connecting subunit having a formula selected from -ZAS*-RL- and –ZAS*-W-RL-, wherein S* has the following formula:

[0639]

[0640] The subscript n is an integer between 2 and 36.

[0641] 55. A camptothecin conjugate according to any one of embodiments 37-49, wherein -ZA- has the following formula:

[0642]

[0643] The succinimide ring in hydrolyzed form is selected as the succinamide moiety, wherein the wavy line marked with a double asterisk (**) indicates the site covalently linked to S*; and the wavy line marked with a triple star (***) indicates the point covalently linked to the sulfur atom of L.

[0644] 56. A camptothecin conjugate according to any one of embodiments 37-49, wherein Q is a linker unit having the formula –ZAS*-W- or –ZASW-RL-, wherein RL is not a glucuronide unit, and wherein –ZAS*-W- in either formula has the following formula:

[0645]

[0646] The succinamide moiety is optionally defined as a succinimide ring in a hydrolyzed form, wherein the subscript n is an integer from 2 to 10; a wavy line marked with a double asterisk (**) indicates a site covalently bonded to D or RL; and a wavy line marked with a triple star (***) indicates a site covalently bonded to the sulfur atom of L.

[0647] 57. Camptothecin conjugate according to implementation scheme 56, wherein the subscript n is an integer from 2 to 4.

[0648] 58. The camptothecin conjugate or its salt according to embodiment 49, wherein –QD has the following structure:

[0649]

[0650] It may optionally have a succinimide ring in a hydrolyzed form as the succinamide moiety, where the wavy line indicates the point covalently connected to the sulfur atom of the ligand unit.

[0651] 59. The camptothecin conjugate or its salt according to embodiment 49, wherein –QD has the following structure:

[0652]

[0653] It may optionally have a succinimide ring in a hydrolyzed form as the succinamide moiety, where the wavy line indicates the point covalently connected to the sulfur atom of the ligand unit.

[0654] 60. A camptothecin conjugate according to any one of implementation schemes 37-59, wherein L is an antibody.

[0655] 61. The camptothecin conjugate according to embodiment 60, wherein the antibody specifically binds to an antigen selected from CD19, CD30, CD33, CD70 and LIV-1.

[0656] 62. A method having the following properties selected from formulas Z'-A-RL-D(i), Z'-A-RL-YD(ii), and Z'-AS * -RL-D(iii), -Z'-AS*-RL-YD(iv), Z'-AB(S * )-RL-D(v), Z'-AB(S * Camptothecin-linker compounds of Z'-RL-YD(vi), Z'-AD(vii), Z'-AS*-WD(viii), Z'-AB(S*)-WD(ix), Z'-AS*-W-RL-D(x) and Z'-AB(S*)-W-RL-D(xi), wherein in each formula Z' is the extended subunit precursor; A is the bond or linker unit; B is the parallel linker unit; S * is a distributing agent; RL is a releasable linker; Y is a spacer subunit; and D is a camptothecin compound selected from the following CPT1, CPT2, CPT3, CPT4, CPT5, CPT6 and CPT7:

[0657]

[0658]

[0659] Where R BThe derivative is selected from -H, C1-C8 alkyl, C1-C8 haloalkyl, C3-C8 cycloalkyl, (C3-C8 cycloalkyl)-C1-C4 alkyl, phenyl, and phenyl-C. 1- Members of C4 alkyl groups; R C Selected from C1-C6 alkyl and C 3- C6 cycloalkyl moiety; R F and R F’ Each is independently selected from -H, C1-C8 alkyl, C1-C8 hydroxyalkyl, C1-C8 aminoalkyl, (C1-C4 alkylamino)-C1-C8 alkyl, N,N-(C1-C4 hydroxyalkyl)(C1-C4 alkyl)amino-C1-C8 alkyl, N,N-bis(C1-C4 alkyl)amino-C 1- C8 alkyl, N-C1-C4 hydroxyalkyl-C1-C8 aminoalkyl, C1-C8 alkyl C(O)-, C1-C8 hydroxyalkyl-C(O)-, C1-C8 aminoalkyl-C(O)-, C3-C 10 cycloalkyl, (C3-C 10 Cycloalkyl)-C1-C4 alkyl, C3-C 10 Heterocyclic alkyl groups, (C3-C 10 Heterocyclic alkyl)-C1-C4 alkyl, phenyl, phenyl-C1-C4 alkyl, diphenyl-C1-C4 alkyl, heteroaryl and heteroaryl-C1-C4 alkyl portions; or R F and R F’ It combines with the nitrogen atoms to form a 5-, 6-, or 7-membered ring having 0 to 3 substituents selected from halogens, C1-C4 alkyl groups, -OH, -OC1-C4 alkyl groups, -NH2, and -NHC. 1- C4 alkyl and -N(C1-C4 alkyl)2; and wherein R B R C R F and R F’ The cycloalkyl, heterocycloalkyl, phenyl, and heteroaryl moieties are composed of 0 to 3 elements selected from halogens, C 1- C4 alkyl, -OH, -OC1-C4 alkyl, -NH2, -NHC 1- C4 alkyl and -N(C 1- Substituents of C4 alkyl group 2; or

[0660] D is a pharmaceutical unit selected from 13a-13c of Table H, 14a-14z of Table I and 18a-18r of Table J.

[0661] And where Q is -ZA-RL-, -ZA-RL-Y-, -ZAS * -RL-、-ZAB(S * -RL-、-ZAS* -RL-Y- or -ZAB(S) * When )-RL-Y-, the covalent bonding site of D is any one of the hydroxyl, aliphatic primary amino, or secondary amino substituents in any one of CPT1, CPT2, CPT3, CPT4, CPT5, CPT6, and CPT7, or any one of the heteroatoms in any one of the substituents in 13a-13c of Table H, 14a-14z of Table I, and 18a-18r of Table J.

[0662] Where Q is -ZA- or -ZAS * -W- or -ZAB(S * -W-, or when Q is -ZAS * -RL-、-ZAB(S * -RL-、-ZAS * -W-RL- or -ZAB(S * When RL is a releasable unit other than a glucuronide unit, the covalent linker of D is the oxygen atom of the hydroxyl substituent in the lactone ring of any one of CPT1, CPT2, CPT3, CPT4, CPT5, CPT6, or CPT7, or any one of 13a-13c in Table H, 14a-14z in Table I, and 18a-18r in Table J; and

[0663] The condition is that when the covalent linkage site is the nitrogen atom of the amino substituent in CPT6, R F and R F’ At least one of them is -H, and the condition is that when D is a CPT1 having a nitrogen atom covalently linked by its amino substituent, the Z'-A- of the camptothecin-linker compounds of formulas (i), (ii), (iii), (iv), (v) and (vi) is not a succinimide-hexanoyl-β-alanyl moiety.

[0664] 63. The camptothecin-linker compound according to embodiment 62 has a formula selected from formula (i), formula (ii), formula (iii), formula (iv), formula (v) and formula (vi), wherein A is a linker unit and RL is a glucuronide unit.

[0665] 64. The camptothecin-linker compound according to embodiment 62 or 63, wherein the covalent linking point of D is an oxygen atom of a hydroxyl substituent on the lactone ring of CPT1, CPT2, CPT3, CPT4, CPT5, CPT6 or CPT7.

[0666] 65. The camptothecin-linker compound according to embodiment 62 or 63, wherein the covalent linking point of D is the oxygen atom of a hydroxyl substituent on the lactone ring of any one of Table H 13a-13c, Table I 14a-14z and Table J 18a-18r.

[0667] 66. The camptothecin-linker compound according to embodiment 62 or 63, wherein the covalent linking site of D is through R of CPT6. F Or R F’ The oxygen atom of the hydroxyl substituent, wherein R F and R F’ At least one of them is C1-C8 hydroxyalkyl N,N-(C1-C4 hydroxyalkyl)(C1-C4 alkyl)-amino-C1-C8 alkyl-, N-C1-C4 hydroxyalkyl-C1-C8 aminoalkyl-, or C1-C8 alkyl C(O)-.

[0668] 67. The camptothecin-linker compound according to embodiment 62 or 63, wherein the covalent linking site of D is through R of CPT6. F Or R F’ The nitrogen atom of the amine substituent, where R F and R F’ At least one of them is C1-C8 aminoalkyl, (C1-C4 alkylamino)-C1-C8 alkyl-, N-(C1-C4 hydroxyalkyl)-C1-C8 aminoalkyl- or C1-C8 aminoalkylC(O)-.

[0669] 68. Camptothecin-linker compound according to embodiment 62 or 63, wherein the covalent linking point of D is the nitrogen atom of the amino substituent of CPT1, provided that -ZA- is not maleimide-hexanoyl-β-alanyl.

[0670] 69. The camptothecin-linker compound according to embodiment 62 or 63, wherein the linking point of D is the nitrogen atom of the amino substituent of CPT4.

[0671] 70. A camptothecin-linker compound according to embodiment 62 or 63, wherein the linking point of D is through the nitrogen atom of the substituent on CPT6, and the condition is R F and R F’ At least one of them is -H.

[0672] 71. The camptothecin-linker compound according to any one of embodiments 62-70, having formula (i) or formula (ii).

[0673] 72. The camptothecin-linker compound according to any one of embodiments 62-70, having formula (iii) or formula (iv).

[0674] 73. The camptothecin-linker compound according to any one of embodiments 62-70, having formula (v) or formula (vi).

[0675] 74. The camptothecin-linker compound according to any one of embodiments 62-70, having formula (i).

[0676] 75. A camptothecin-linker compound according to any one of embodiments 62-70, having formula (ii).

[0677] 76. A camptothecin-linker compound according to any one of embodiments 62-64, 66, 67 and 70-75, wherein D is CPT6.

[0678] 77. A camptothecin-linker compound according to any one of embodiments 62-64, 69 and 71-75, wherein D is CPT4.

[0679] 78. A camptothecin-linker compound according to any one of embodiments 62-64 and 71-75, wherein D is selected from CPT1, CPT2, CPT3 and CPT5.

[0680] 79. A camptothecin-linker compound according to any one of embodiments 62-78, wherein Z' is a maleimide moiety.

[0681] 80. A camptothecin-linker compound according to any one of embodiments 62-78, wherein Z'-A- is maleimide propionyl, maleimide propionyl-β-alanyl or mDPR, and its basic nitrogen is optionally protonated or protected by an acid-insecure protecting matrix, provided that Z'-A- is not a maleimide-hexanoyl-β-alanyl moiety when D is a CPT1 having a nitrogen atom covalently linked through its amino substituent.

[0682] 81. A camptothecin-linker compound according to any one of embodiments 62-70, having formulas (iii), (iv), (v), and (vi), wherein S * It is a PEG group.

[0683] 82. A camptothecin-linker compound according to any one of embodiments 63-81, wherein RL is a glucuronide unit having the following structure:

[0684]

[0685] A wavy line marked with a single asterisk (*) indicates a site covalently connected to D or the spacer subunit (Y); and a wavy line marked with a double asterisk (**) indicates a site covalently connected to A, B, or S. * Points that are covalently connected.

[0686] 83. The camptothecin-linker compound according to embodiment 82, having formula (ii), formula (iv) or formula (vi), wherein the spacer subunit (Y) has the following formula:

[0687]

[0688] Where EWG is an electron-withdrawing group; O* represents the oxygen atom from the hydroxyl functional group of D; the wavy line adjacent to the nitrogen atom indicates the site covalently bonded to the carbonyl carbon atom of the glucuronide unit; and the wavy line adjacent to O* indicates the site covalently bonded to the rest of D.

[0689] 84. The camptothecin-linker compound according to embodiment 82, having formula (ii), formula (iv) or formula (vi), wherein D is selected from R. F and R F’ Each of them is -H CPT1, CPT4 and CPT6, and the spacer subunit (Y) has the following formula:

[0690]

[0691] Where EWG is an electron-withdrawing group; the wavy line adjacent to the nitrogen atom indicates the site covalently bonded to the carbonyl carbon atom of the glucuronide unit; and the wavy line adjacent to the carbonyl carbon atom indicates the site covalently bonded to the R atom. F and R F’ Each of these is a site where the nitrogen atom of an amino substituent of CPT1, CPT4, or CPT6 is covalently linked.

[0692] 85. The camptothecin-linker compound according to embodiment 82, having formula (ii), formula (iv) or formula (vi), wherein D is selected from R. F and R F’ Each of them is -H CPT1, CPT4 and CPT6, and the spacer subunit (Y) has the following formula:

[0693]

[0694] Where EWG is an electron-withdrawing group; the wavy line adjacent to the nitrogen atom indicates the site covalently bonded to the carbonyl carbon atom of the glucuronide unit; and the wavy line adjacent to the carbonyl carbon atom indicates the site covalently bonded to the R atom. F and R F’ Each of these is a site where the nitrogen atom of an amino substituent of CPT1, CPT4, or CPT6 is covalently linked.

[0695] 86. A camptothecin-linker compound according to any one of embodiments 62-85, wherein A is composed of an alkynyl moiety capable of 1,3-dipolar cycloaddition with an azide substituent from a chemically modified targeting agent that is a precursor of a ligand unit of the camptothecin conjugate, thereby providing a conjugate having a linker unit composed of a triazole moiety.

[0696] 87. A camptothecin-linker compound according to any one of embodiments 62-85, wherein Z'-A- is composed of a maleimide-alkanoyl moiety or a maleimide and triazole moiety, provided that Z'-A- is composed of a maleimide and triazole moiety when CPT1 has a covalently linked nitrogen atom through its amino substituent.

[0697] 88. A camptothecin-linker compound according to any one of embodiments 62-85, wherein Z'-A- is composed of a maleimide-alkanoyl-β-alanyl moiety, provided that when D is CPT1, D has a covalent connection to the oxygen atom of the hydroxyl substituent on its lactone ring.

[0698] 89. A camptothecin-linker compound according to any one of embodiments 58-79, wherein Z'-A- is composed of mDPR, and its basic nitrogen is optionally protonated or protected by an acid-instable protecting matrix.

[0699] 90. The camptothecin-linker compound according to embodiment 62, having formula (vii), formula (viii) or formula (ix), wherein A is a linker unit, or having formula (i), formula (iii), formula (x) or formula (xi), wherein A is a linker unit and RL is a releasable linker other than a glucuronide unit.

[0700] 91. The camptothecin-linker compound according to embodiment 62 or 90, having formula (viii) or formula (x).

[0701] 92. The camptothecin-linker compound according to embodiment 62 or 90, having formula (i) or formula (iii).

[0702] 93. The camptothecin-linker compound according to embodiment 62 or 90 has formula (vii).

[0703] 94. A camptothecin-linker compound according to any one of embodiments 62 and 90-93, wherein D has the formula CPT2.

[0704] 95. A camptothecin-linker compound according to any one of embodiments 62 and 90-93, wherein D has the formula CPT3.

[0705] 96. A camptothecin-linker compound according to any one of embodiments 62 and 90-93, wherein D has the formula CPT1.

[0706] 97. A camptothecin-linker compound according to any one of embodiments 62 and 90-93, wherein D has the formula CPT4.

[0707] 98. A camptothecin-linker compound according to any one of embodiments 62 and 90-93, wherein D has the formula CPT5.

[0708] 99. A camptothecin-linker compound according to any one of embodiments 62 and 90-93, wherein D has the formula CPT6.

[0709] 100. A camptothecin-linker compound according to any one of embodiments 62 and 90-99, having formula (i), (iii), or (x), wherein RL has the following formula:

[0710]

[0711] The wavy lines marked with double asterisks (**) indicate sites covalently connected to D; and the wavy lines marked with single asterisks (*) indicate sites covalently connected to A, S*, or W.

[0712] 101. The camptothecin-linker compound according to embodiment 100, having formula (x), wherein W is an amino acid unit selected from N-methyl-glycine (sarcosine), N-methyl-alanine, N-methyl-β-alanine, valine and N-methyl-valine.

[0713] 102. A camptothecin-linker compound according to any one of embodiments 62-101, wherein Z'-A- is composed of a maleimide-alkanoyl moiety, a maleimide group and a triazole moiety or mDPR, and its basic nitrogen atom is optionally protonated or protected by an acid-instable protecting matrix.

[0714] 102. The camptothecin-linker compound according to any one of embodiments 62-101, wherein Z'-A- is composed of a maleimide-alkanoyl moiety.

[0715] 103. The camptothecin-linker compound according to any one of embodiments 62-101, wherein Z'-A- is composed of mDPR.

[0716] 104. A camptothecin-linker compound according to any one of embodiments 62-70, having formula (iii) or formula (x), wherein Z'-A- has a formula selected from:

[0717]

[0718] The wavy lines marked with double asterisks (**) indicate sites covalently connected to S*.

[0719] 105. A camptothecin-linker compound according to any one of embodiments 62-70 and 94-104, having formula (iii) or formula (x), wherein S* has the following formula:

[0720]

[0721] The subscript n is an integer between 2 and 36.

[0722] 106. A camptothecin-linker compound according to any one of embodiments 62-70 and 94-104, wherein Z'-A- has the following formula:

[0723]

[0724] The wavy lines marked with double asterisks (**) indicate sites covalently connected to S*.

[0725] 107. A camptothecin-linker compound according to any one of embodiments 62-70 and 94-105, having formula (viii) or formula (x), wherein Z'-AS*-W- has the following formula:

[0726]

[0727] The subscript n is an integer from 2 to 10; the wavy line marked with a double asterisk (**) indicates a site covalently bonded to D or RL; and the wavy line marked with a triple star (***) indicates a site covalently bonded to the sulfur atom of L.

[0728] 108. The camptothecin-linker compound according to embodiment 107, wherein the subscript n is an integer from 2 to 4.

[0729] 109. The camptothecin-linker compound or its salt according to embodiment 62 has the following structure:

[0730]

[0731] 110. The camptothecin-linker compound or its salt according to embodiment 62 has the following structure:

[0732]

[0733] 111. The camptothecin-linker compound or its salt according to embodiment 62 has the following structure:

[0734]

[0735] 112. The camptothecin-linker compound or its salt according to embodiment 62 has the following structure:

[0736]

[0737] 113. The camptothecin-linker compound or its salt according to embodiment 62 has the following structure:

[0738]

[0739] 114. Use of camptothecin conjugates in the preparation of medicaments for treating cancer in a subject, wherein the camptothecin conjugate has the formula of any one of embodiments 1-113.

[0740] 115. The use according to embodiment 114, wherein the cancer is selected from lymphoma, leukemia and solid tumors.

[0741] 116. The use according to embodiment 114, wherein the cancer is lymphoma or leukemia.

[0742] 117. Use of camptothecin conjugates in the preparation of a medicament for treating an autoimmune disease in a subject, wherein the camptothecin conjugate has the formula of any one of embodiments 1-113, and the autoimmune disease is selected from Th2 lymphocyte-associated diseases, Th1 lymphocyte-associated diseases, and activated B lymphocyte-associated diseases.

[0743] 118. A method for preparing a camptothecin conjugate of any one of embodiments 1-61, the method comprising the step of contacting an antibody having a functional group reactive to the Z' of a camptothecin-linker compound of any one of embodiments 62-113.

[0744] 1A. A method having the formula L-(QD) p Camptothecin conjugates or salts thereof, wherein L is a ligand unit; Q is a linker unit having a formula selected from: -ZA-RL-, -ZA-RL-Y-, -ZAS * -RL-、-ZAB(S * -RL-, -ZAS*-RL-Y-, and –ZAB(S*)-RL-Y-, where Z is an extended sub-element; A is a key or connector element; B is a parallel connector element; S * For dispensing agents; RL for glucuronide units; and Y for spacer units; D for pharmaceutical units selected from the following:

[0745]

[0746] Where R BThe derivative is selected from H, C1-C8 alkyl, C1-C8 haloalkyl, C3-C8 cycloalkyl, (C3-C8 cycloalkyl)-C1-C4 alkyl, phenyl, and phenyl-C 1- Members of C4 alkyl groups; R C Selected from C1-C6 alkyl and C 3- Members of C6 cycloalkyl groups; R F and R F’ Each is independently selected from -H, C1-C8 alkyl, C1-C8 hydroxyalkyl, C1-C8 aminoalkyl, (C1-C4 alkylamino)-C1-C8 alkyl, N,N-(C1-C4 hydroxyalkyl)(C1-C4 alkyl)amino-C1-C8 alkyl, N,N-bis(C1-C4 alkyl)amino-C 1- C8 alkyl, N-C1-C4 hydroxyalkyl-C1-C8 aminoalkyl, C1-C8 alkyl C(O)-, C1-C8 hydroxyalkyl-C(O)-, C1-C8 aminoalkyl-C(O)-, C3-C 10 cycloalkyl, (C3-C 10 Cycloalkyl)-C1-C4 alkyl, C3-C 10 Heterocyclic alkyl groups, (C3-C 10 Members of heterocyclic alkyl (C1-C4 alkyl), phenyl, phenyl-C1-C4 alkyl, diphenyl-C1-C4 alkyl, heteroaryl and heteroaryl-C1-C4 alkyl; or R F and R F’ It combines with the nitrogen atoms to form a 5-, 6-, or 7-membered ring having 0 to 3 substituents selected from halogens, C1-C4 alkyl groups, -OH, -OC1-C4 alkyl groups, -NH2, and -NHC. 1- C4 alkyl and -N(C1-C4 alkyl)2; and wherein R B R C R F and R F’ The cycloalkyl, heterocycloalkyl, phenyl, and heteroaryl moieties are composed of 0 to 3 elements selected from halogens, C 1- C4 alkyl, -OH, -OC1-C4 alkyl, -NH2, -NHC 1- C4 alkyl and -N(C 1- Substituents of C4 alkyl)2; subscript p is an integer from 1 to 16; and

[0747] Where Q is linked by any hydroxyl and amino group present on CPT1, CPT2, CPT3, CPT4, CPT5, or CPT6; and where -ZA- is not maleimide-hexanoyl-β-alanyl when D is CPT1, linked by an amino group of CPT1.

[0748] 2A. A camptothecin conjugate according to embodiment 1A, wherein Q is a connecting subunit having a formula selected from -ZA-RL- and -ZA-RL-Y-.

[0749] 3A. A camptothecin conjugate according to embodiment 1A, wherein Q is a compound having a composition selected from -ZAS. * -RL- and -ZAS * -RL-Y- style connection subunit.

[0750] 4A. A camptothecin conjugate according to embodiment 1A, wherein Q is a connecting subunit having a formula selected from -ZAB(S*)-RL- and –ZAB(S*)-RL-Y-.

[0751] 5A. A camptothecin conjugate according to any one of embodiments 1A-4A, wherein D has the formula CPT1.

[0752] 6A. A camptothecin conjugate according to any one of embodiments 1A-4A, wherein D has the formula CPT2.

[0753] 7A. A camptothecin conjugate according to any one of embodiments 1A-4A, wherein D has the formula CPT3.

[0754] 8A. A camptothecin conjugate according to any one of embodiments 1A-4A, wherein D has the formula CPT4.

[0755] 9A. A camptothecin conjugate according to any one of embodiments 1A-4A, wherein D has the formula CPT5.

[0756] 10A. A camptothecin conjugate according to any one of embodiments 1A-4A, wherein D has the formula CPT6.

[0757] 11A. A camptothecin conjugate according to any one of embodiments 1A-4A, wherein L is an antibody.

[0758] 12A. A camptothecin conjugate according to embodiment 1A, wherein Q comprises a glucuronide unit (RL) having the following formula:

[0759]

[0760] Sugar is a hexose form of a natural or non-natural monosaccharide, and RL is linked to a primary amine of any one of CPT1, CPT4, or CPT6; a wavy line marked with a single asterisk (*) indicates a site linked to a primary amine of CPT1, CPT4, or CPT6 or to a spacer subunit (Y), and a wavy line marked with a ** indicates a site linked to other linker components of Q.

[0761] 13A. A camptothecin conjugate according to embodiment 12A, wherein a spacer subunit (Y) is present and comprises:

[0762]

[0763] EWG is an electron-withdrawing group.

[0764] 14A. Camptothecin conjugate according to embodiment 12A, wherein A comprises a triazole formed from an alkyne and an azide when using knock chemistry.

[0765] 15A. A camptothecin conjugate according to embodiment 12A, wherein -ZA- comprises a maleimide-alkanoic acid component, a maleimide group and a triazole component, or an mDPR component.

[0766] 16A. A camptothecin conjugate according to embodiment 12A, wherein -ZA- comprises a maleimide-alkanoyl-β-alanyl group.

[0767] 17A. Camptothecin conjugate according to embodiment 12A, wherein ZA- comprises the mDPR component.

[0768] 18A. The camptothecin conjugate according to embodiment 12A, wherein Q has the following formula:

[0769]

[0770] The wavy lines marked with a single asterisk (*) indicate sites connected to primary amines of CPT1, CPT4, or CPT6, or to spacer subunits, while the wavy lines marked with a *** indicate sites connected to sulfur atoms of L.

[0771] 19A. The camptothecin conjugate according to embodiment 1A, wherein the glucuronic acid unit has the following formula:

[0772]

[0773] The wavy lines marked with a single asterisk (*) indicate points connected to the hydroxyl groups of CPT1, CPT2, CPT3, CPT4, CPT5, or CPT6, or to the spacer subunit (Y); and the wavy lines marked with a double asterisk (**) indicate points connected to A, B, S. * Or the points connected by Z.

[0774] 20A. A camptothecin conjugate according to embodiment 19A, wherein a spacer subunit is present and comprises:

[0775]

[0776] EWG is an electron-withdrawing group.

[0777] 21A. A camptothecin conjugate according to embodiment 19A, wherein a spacer subunit is present and comprises:

[0778]

[0779] 22A. Camptothecin conjugate according to embodiment 19A, wherein A comprises a triazole formed from an alkyne and an azide when using knock chemistry.

[0780] 23A. A camptothecin conjugate according to embodiment 19A, wherein -ZA- comprises a maleimide-alkanoic acid component, a maleimide group and a triazole component, or an mDPR component.

[0781] 24A. A camptothecin conjugate according to embodiment 19A, wherein -ZA- contains a maleimide-alkanoyl-β-alanyl group.

[0782] 25A. Camptothecin conjugate according to embodiment 19A, wherein -ZA- comprises the mDPR component.

[0783] 26A. A camptothecin conjugate according to embodiment 19A, wherein -ZA- comprises a maleimide propionyl group and a dispensing agent (S * It exists in the connecting subunit.

[0784] 27A. The camptothecin conjugate according to embodiment 19A, wherein Q has the following formula:

[0785]

[0786] Where n is an integer from 1 to 50, a wavy line marked with a single asterisk (*) indicates a site connected to D or to a spacer subunit (Y); and a wavy line marked with a *** indicates a site connected to a sulfur atom of L.

[0787] 28A. Camptothecin conjugate according to implementation scheme 27A, wherein n is 4.

[0788] 29A. A camptothecin conjugate according to any one of embodiments 1A-28A, wherein L is an antibody that specifically binds to an antigen selected from CD19, CD30, CD33, CD70 and LIV-1.

[0789] 30A. A method having the following properties selected from Z'-A-RL-D(i), Z'-A-RL-YD(ii), Z'-AS * -RL-D(iii), -Z'-AS*-RL-YD(iv), Z'-AB(S * )-RL-D(v) and Z'-AB(S *The camptothecin-linker compound of the formula )-RL-YD(vi), where Z' is the extended subunit; A is the bond or linker unit; B is the parallel linker unit; S * The partitioning agent is RL, which is a glucuronide unit; Y is a spacer unit; and D is a camptothecin compound selected from the following:

[0790]

[0791]

[0792] Where R B The derivative is selected from H, C1-C8 alkyl, C1-C8 haloalkyl, C3-C8 cycloalkyl, (C3-C8 cycloalkyl)-C1-C4 alkyl, phenyl, and phenyl-C 1- Members of C4 alkyl groups; R C Selected from C1-C6 alkyl and C 3- Members of C6 cycloalkyl groups; R F and R F’ Each is independently selected from -H, C1-C8 alkyl, C1-C8 hydroxyalkyl, C1-C8 aminoalkyl, (C1-C4 alkylamino)-C1-C8 alkyl, N,N-(C1-C4 hydroxyalkyl)(C1-C4 alkyl)amino-C1-C8 alkyl, N,N-bis(C1-C4 alkyl)amino-C 1- C8 alkyl, N-C1-C4 hydroxyalkyl-C1-C8 aminoalkyl, C1-C8 alkyl C(O)-, C1-C8 hydroxyalkyl-C(O)-, C1-C8 aminoalkyl-C(O)-, C3-C 10 cycloalkyl, (C3-C 10 Cycloalkyl)-C1-C4 alkyl, C3-C 10 Heterocyclic alkyl groups, (C3-C 10 Members of heterocyclic alkyl (C1-C4 alkyl), phenyl, phenyl-C1-C4 alkyl, diphenyl-C1-C4 alkyl, heteroaryl and heteroaryl-C1-C4 alkyl; or R F and R F’ It combines with the nitrogen atoms to form a 5-, 6-, or 7-membered ring having 0 to 3 substituents selected from halogens, C1-C4 alkyl groups, -OH, -OC1-C4 alkyl groups, -NH2, and -NHC. 1- C4 alkyl and -N(C1-C4 alkyl)2; and wherein R B R C R F and R F’ The cycloalkyl, heterocycloalkyl, phenyl, and heteroaryl moieties are composed of 0 to 3 elements selected from halogens, C 1- C4 alkyl, -OH, -OC1-C4 alkyl, -NH2, -NHC1- C4 alkyl and -N(C 1- Substituents of C4 alkyl)2;

[0793] The subscript p is an integer from 1 to 16; and where Q is linked by any hydroxyl and amino group present on CPT1, CPT2, CPT3, CPT4, CPT5, or CPT6; and where -ZA- is not maleimide-hexanoyl-β-alanyl when D is CPT1 and linked by an amino group of CPT1.

[0794] 31A. The camptothecin-linker compound according to embodiment 30A, having formula (i) or formula (ii).

[0795] 32A. The camptothecin-linker compound according to embodiment 30A, having formula (iii) or formula (iv).

[0796] 33A. The camptothecin-linker compound according to embodiment 30A, having formula (v) or formula (vi).

[0797] 34A. The camptothecin-linker compound according to embodiment 30A has formula (i).

[0798] 35A. The camptothecin-linker compound according to embodiment 30A has formula (ii).

[0799] 36A. A camptothecin-linker conjugate according to any one of embodiments 30A-34A, wherein D is CPT6.

[0800] 37A. A camptothecin-linker conjugate according to any one of embodiments 30A-34A, wherein D is CPT4.

[0801] 38A. A camptothecin-linker conjugate according to any one of embodiments 30A-34A, wherein D is selected from CPT1, CPT2, CPT3 and CPT5.

[0802] 39A. A camptothecin-linker conjugate according to any one of embodiments 30A-34A, wherein Z' is a maleimide group.

[0803] 40A. A camptothecin-linker conjugate according to any one of embodiments 30A-34A, wherein Z'-A– is maleimide propionyl, mDPR, or maleimide propionyl-β-alanyl.

[0804] 41A. A camptothecin-linker conjugate according to any one of embodiments 30A and 32A-34A, wherein S * It is a PEG group.

[0805] 42A. The camptothecin-linker compound according to embodiment 30A, wherein the glucuronide unit has the following formula:

[0806]

[0807] The wavy lines marked with a single asterisk (*) indicate sites connected to D or spacer units (Y); and the wavy lines marked with a double asterisk (**) indicate sites connected to other linker components of the camptothecin-linker compound, A, B, S. * Or the points connected by Z.

[0808] 42A. A camptothecin-linker compound according to embodiment 30A, wherein a spacer subunit is present and comprises:

[0809]

[0810] EWG is an electron-withdrawing group.

[0811] 43A. A camptothecin-linker compound according to embodiment 42A, wherein a spacer subunit is present and comprises:

[0812]

[0813] 44A. Camptothecin-linker compound according to embodiment 42A, wherein A comprises a triazole formed from an alkyne and an azide when using knock chemistry.

[0814] 45A. Camptothecin-linker compound according to embodiment 42A, wherein Z'-A- comprises a maleimide-alkanoic acid component, a maleimide group and a triazole component, or an mDPR component.

[0815] 46A. Camptothecin-linker compound according to embodiment 42A, wherein Z'-A- comprises maleimide-alkanoyl-β-alanyl group.

[0816] 47A. Camptothecin-linker compound according to embodiment 42A, wherein Z'-A- comprises the mDPR component.

[0817] 48A. A camptothecin-linker compound according to embodiment 42A, wherein Z'-A- comprises a maleimide propionyl group and a dispensing agent (S * It exists in the connecting subunit.

[0818] 49A. A camptothecin-linker compound according to embodiment 30A, wherein formulas (i) and (ii) comprise the following:

[0819]

[0820] The wavy lines marked with a single asterisk (*) indicate sites connected to primary amines of CPT1, CPT4, or CPT6, or to spacer subunits.

[0821] 50A. A method for treating cancer in a subject in need, the method comprising administering to the subject a camptothecin conjugate of any one of embodiments 1A-29A.

[0822] 51A. The method according to embodiment 50A, wherein the cancer is selected from lymphoma, leukemia and solid tumors.

[0823] 52A. The method according to embodiment 50A, wherein the cancer is lymphoma or leukemia.

[0824] 53A. The method according to any one of embodiments 50A-53A further includes an additional therapeutic agent.

[0825] 54A. The method according to embodiment 53A, wherein the additional therapeutic agent is one or more chemotherapeutic agents or radiotherapy.

[0826] 55A. A method for treating an autoimmune disease in a subject in need, the method comprising administering to the subject a camptothecin conjugate of any one of embodiments 1A-29A.

[0827] 56A. The method according to embodiment 55A, wherein the autoimmune disease is selected from Th2 lymphocyte-associated diseases, Th1 lymphocyte-associated diseases, and activated B lymphocyte-associated diseases.

[0828] 57A. A method for preparing a camptothecin conjugate of any one of embodiments 1A-29A, said method comprising reacting an antibody with a camptothecin-linker compound of any one of embodiments 39A-49A.

[0829] 58A. A kit comprising a camptothecin conjugate of any one of embodiments 1A-29A.

[0830] 59A. The kit according to embodiment 58A further contains additional therapeutic agents.

[0831] 1B. A method having the formula L-(QD) p The camptothecin conjugate or its salt, wherein L is a ligand unit; Q is a linker unit having a formula selected from: -ZA-, -ZA-RL-, -ZAS*-W-, -ZAB(S*)-W-, -ZAS*-RL-, -ZAB(S*)-RL-, -ZAS*-W-RL-, and -ZAB(S*)-W-RL-, wherein Z is an extension unit; A is a bond or linker unit; B is a parallel linker unit; S* is a dispensing agent; RL is a releasable linker; and W is an amino acid unit; D is a pharmaceutical unit selected from the following:

[0832]

[0833] Where R B The derivative is selected from H, C1-C8 alkyl, C1-C8 haloalkyl, C3-C8 cycloalkyl, (C3-C8 cycloalkyl)-C1-C4 alkyl, phenyl, and phenyl-C 1- Members of C4 alkyl groups; R C Selected from C1-C6 alkyl and C 3- Members of C6 cycloalkyl groups; R F and R F’ Each is independently selected from -H, C1-C8 alkyl, C1-C8 hydroxyalkyl, C1-C8 aminoalkyl, (C1-C4 alkylamino)-C1-C8 alkyl, N,N-(C1-C4 hydroxyalkyl)(C1-C4 alkyl)amino-C1-C8 alkyl, N,N-bis(C1-C4 alkyl)amino-C 1- C8 alkyl, N-C1-C4 hydroxyalkyl-C1-C8 aminoalkyl, C1-C8 alkyl C(O)-, C1-C8 hydroxyalkyl-C(O)-, C1-C8 aminoalkyl-C(O)-, C3-C 10 cycloalkyl, (C3-C 10 Cycloalkyl)-C1-C4 alkyl, C3-C 10 Heterocyclic alkyl groups, (C3-C 10 Members of heterocyclic alkyl (C1-C4 alkyl), phenyl, phenyl-C1-C4 alkyl, diphenyl-C1-C4 alkyl, heteroaryl and heteroaryl-C1-C4 alkyl; or R F and R F’ It combines with the nitrogen atoms to form a 5-, 6-, or 7-membered ring having 0 to 3 substituents selected from halogens, C1-C4 alkyl groups, -OH, -OC1-C4 alkyl groups, -NH2, and -NHC. 1- C4 alkyl and -N(C1-C4 alkyl)2; and wherein R B R C R F and R F’ The cycloalkyl, heterocycloalkyl, phenyl, and heteroaryl moieties are composed of 0 to 3 elements selected from halogens, C 1- C4 alkyl, -OH, -OC1-C4 alkyl, -NH2, -NHC 1- C4 alkyl and -N(C 1- Substituents of C4 alkyl group 2; and

[0834] The connection point between D and Q is through the oxygen atom of the hydroxyl substituent of the lactone ring of CPT1, CPT2, CPT3, CPT4, CPT5, or CPT6.

[0835] 2B. Camptothecin conjugate according to embodiment 1B, wherein Q is a compound having the formula -ZAS * -W- connecting subunit.

[0836] 3B. Camptothecin conjugate according to embodiment 1B, wherein Q is a compound having the formula -ZAS * -W-RL- connecting subunit.

[0837] 4B. Camptothecin conjugate according to embodiment 1B, wherein Q is a linker unit having the formula -ZA-.

[0838] 5B. A camptothecin conjugate according to any one of implementation schemes 1B-4B, wherein D has the formula CPT2.

[0839] 6B. A camptothecin conjugate according to any one of implementation schemes 1B-4B, wherein D has the formula CPT3.

[0840] 7B. A camptothecin conjugate according to any one of embodiments 1B-4B, wherein D has the formula CPT1.

[0841] 8B. A camptothecin conjugate according to any one of embodiments 1B-4B, wherein D has the formula CPT4.

[0842] 9B. A camptothecin conjugate according to any one of embodiments 1B-4B, wherein D has the formula CPT5.

[0843] 10B. A camptothecin conjugate according to any one of embodiments 1B-4B, wherein D has the formula CPT6.

[0844] 11B. A camptothecin conjugate according to any one of implementation schemes 1B-4B, wherein L is an antibody.

[0845] 12B. The camptothecin conjugate according to embodiment 3B, wherein RL has the following formula:

[0846]

[0847] The wavy lines marked with ** represent points connected to D, and the wavy lines marked with * represent points connected to another component of Q.

[0848] 13B. The camptothecin conjugate according to embodiment 12B, wherein W is an amino acid unit selected from N-methylglycine, N-methylalanine, N-methyl-β-alanine, valine, and N-methylvaline.

[0849] 14B. A camptothecin conjugate according to embodiment 12B, wherein -ZA- comprises a maleimide-alkanoyl moiety, or a maleimide and triazole moiety, or an mDPR moiety.

[0850] 15B. A camptothecin conjugate according to embodiment 12B, wherein -ZA- comprises a maleimide-alkanoyl moiety.

[0851] 16B. The camptothecin conjugate according to embodiment 12B, wherein -ZA- has a formula selected from:

[0852]

[0853] The wavy lines marked with ** indicate sites connected to S*; and the wavy lines marked with *** indicate sites connected to sulfur atoms of L.

[0854] 17B. The camptothecin conjugate according to embodiment 12B, wherein S* has the following formula:

[0855]

[0856] And the index n is an integer between 2 and 36.

[0857] 18B. A camptothecin conjugate according to embodiment 12B, wherein W is an amino acid unit selected from N-methylglycine, N-methylalanine, N-methyl-β-alanine, valine, and N-methylvaline.

[0858] 19B. A camptothecin conjugate according to embodiment 12B, wherein -ZA- comprises a maleimide-alkanoyl moiety, or a maleimide and triazole moiety, or an mDPR moiety.

[0859] 20B. Camptothecin conjugate according to embodiment 2B, wherein -ZA- comprises a maleimide-alkanoyl moiety.

[0860] 21B. The camptothecin conjugate according to embodiment 2B, wherein -ZA- has a formula selected from the following:

[0861]

[0862] The wavy lines marked with ** indicate sites connected to S*; and the wavy lines marked with *** indicate sites connected to sulfur atoms of L.

[0863] 22B. The camptothecin conjugate according to embodiment 2B, wherein S* has the following formula:

[0864]

[0865] 23B. The camptothecin conjugate according to embodiment 18B, wherein -ZA- is

[0866]

[0867] The wavy lines marked with ** indicate sites connected to S*; and the wavy lines marked with *** indicate sites connected to sulfur atoms of L.

[0868] 24B. The camptothecin conjugate according to embodiment 18B, wherein Q has the following formula:

[0869]

[0870] Where n is an integer from 2 to 10; the wavy line marked with ** indicates the site connected to D; and the wavy line marked with *** indicates the site connected to the sulfur atom of L.

[0871] 25B. Camptothecin conjugate according to implementation scheme 27B, wherein n is 2-4.

[0872] 26B. Camptothecin conjugates according to embodiments 1B-25B, wherein L is an antibody that specifically binds to antigens selected from CD19, CD30, CD33, CD70 and LIV-1.

[0873] 27B. A method having the following properties selected from Z'-A-RL-D(i), Z'-A-RL-YD(ii), Z'-AS * -RL-D(iii), -Z'-AS*-RL-YD(iv), Z'-AB(S * )-RL-D(v) and Z'-AB(S * The camptothecin-linker compound of the formula )-W-RL-D, where Z' is the elongation subunit; A is the bond or linker unit; B is the parallel linker unit; S * is a dispensing agent; RL is a releasable linker subunit; and D is a drug unit selected from the following:

[0874]

[0875] Where R B The derivative is selected from H, C1-C8 alkyl, C1-C8 haloalkyl, C3-C8 cycloalkyl, (C3-C8 cycloalkyl)-C1-C4 alkyl, phenyl, and phenyl-C 1- Members of C4 alkyl groups; R C Selected from C1-C6 alkyl and C 3- Members of C6 cycloalkyl groups; R F and R F’Each is independently selected from -H, C1-C8 alkyl, C1-C8 hydroxyalkyl, C1-C8 aminoalkyl, (C1-C4 alkylamino)-C1-C8 alkyl, N,N-(C1-C4 hydroxyalkyl)(C1-C4 alkyl)amino-C1-C8 alkyl, N,N-bis(C1-C4 alkyl)amino-C 1- C8 alkyl, N-C1-C4 hydroxyalkyl-C1-C8 aminoalkyl, C1-C8 alkyl C(O)-, C1-C8 hydroxyalkyl-C(O)-, C1-C8 aminoalkyl-C(O)-, C3-C 10 cycloalkyl, (C3-C 10 Cycloalkyl)-C1-C4 alkyl, C3-C 10 Heterocyclic alkyl groups, (C3-C 10 Members of heterocyclic alkyl-C1-C4 alkyl, phenyl, phenyl-C1-C4 alkyl, diphenyl-C1-C4 alkyl, heteroaryl, and heteroaryl-C1-C4 alkyl; or

[0876] R F and R F’ It combines with the nitrogen atoms to form a 5-, 6-, or 7-membered ring having 0 to 3 substituents selected from halogens, C1-C4 alkyl groups, -OH, -OC1-C4 alkyl groups, -NH2, and -NHC. 1- C4 alkyl and -N(C1-C4 alkyl)2; and wherein R B R C R F and R F’ The cycloalkyl, heterocycloalkyl, phenyl, and heteroaryl moieties are composed of 0 to 3 elements selected from halogens, C 1- C4 alkyl, -OH, -OC1-C4 alkyl, -NH2, -NHC 1- C4 alkyl and -N(C 1- Substituents of C4 alkyl)2; and wherein R B R C R F and R F’ The cycloalkyl, heterocycloalkyl, phenyl, and heteroaryl moieties are composed of 0 to 3 elements selected from halogens, C 1- C4 alkyl, -OH, -OC1-C4 alkyl, -NH2, -NHC 1- C4 alkyl and -N(C 1- Substituents of C4 alkyl group 2; and

[0877] The connection point between D and Q is through the oxygen atom of the hydroxyl substituent of the lactone ring of CPT1, CPT2, CPT3, CPT4, CPT5, or CPT6.

[0878] 28B. The camptothecin-linker compound according to embodiment 27B, having formula (i) or formula (ii).

[0879] 29B. The camptothecin-linker compound according to embodiment 27B, having formula (iii) or formula (iv).

[0880] 30B. The camptothecin-linker compound according to embodiment 27B, having formula (v) or formula (vi).

[0881] 31B. The camptothecin-linker compound according to embodiment 27B has formula (i).

[0882] 32B. The camptothecin-linker compound according to embodiment 27B has formula (ii).

[0883] 33B. A camptothecin conjugate according to any one of implementation schemes 27B-31B, wherein D is CPT2.

[0884] 34B. A camptothecin-linker conjugate according to any one of embodiments 27B-31B, wherein D is CPT3.

[0885] 35B. A camptothecin-linker conjugate according to any one of embodiments 27B-31B, wherein D is selected from CPT1, CPT4, CPT5 and CPT6.

[0886] 36B. A camptothecin-linker conjugate according to any one of embodiments 27B-31B, wherein Z' is a maleimide group.

[0887] 37B. A camptothecin-linker conjugate according to any one of embodiments 27B-31B, wherein Z'-A– is maleimide propionyl, mDPR, or maleimide propionyl-β-alanyl.

[0888] 38B. A camptothecin-linker conjugate according to any one of embodiments 27B and 29B-31B, wherein S * It is a PEG group.

[0889] 39B. Camptothecin-linker conjugate according to embodiment 27B, wherein RL has the following formula:

[0890]

[0891] The wavy lines marked with ** represent points connected to D, and the wavy lines marked with * represent points connected to another component of Q.

[0892] 40B. Camptothecin-linker conjugate according to embodiment 39B, wherein Z'-A- comprises a maleimide-alkanoic acid moiety, or a maleimide and triazole moiety, or an mDPR moiety.

[0893] 41B. Camptothecin-linker conjugate according to embodiment 39B, wherein Z'-A- comprises maleimide-alkanoyl-β-alanyl group.

[0894] 42B. Camptothecin-linker conjugate according to embodiment 39B, wherein Z'-A- comprises the mDPR moiety.

[0895] 43B. A camptothecin-linker conjugate according to embodiment 39B, wherein Z'-A- comprises a maleimide propionyl moiety and wherein a partitioning agent (S) is present. * ).

[0896] 44B. The camptothecin-linker conjugate according to embodiment 39B has the following formula:

[0897]

[0898] 45B. The camptothecin-linker conjugate according to embodiment 39B has the following formula:

[0899]

[0900] 46B. A method of treating cancer in a subject in need, the method comprising administering to the subject a camptothecin conjugate of any one of embodiments 1B-29B.

[0901] 47B. The method according to embodiment 46B, wherein the cancer is selected from lymphoma, leukemia and solid tumors.

[0902] 48B. The method according to implementation scheme 46B, wherein the cancer is lymphoma or leukemia.

[0903] 49B. The method according to any one of embodiments 46B-48B further includes the application of an additional therapeutic agent.

[0904] 50B. The method according to embodiment 39B, wherein the additional therapeutic agent is one or more chemotherapeutic agents or radiotherapy.

[0905] 51B. A method for treating an autoimmune disease in a subject in need, the method comprising administering to the subject a camptothecin conjugate of any one of embodiments 1B-26B.

[0906] 52B. The method according to embodiment 51B, wherein the autoimmune disease is selected from Th2 lymphocyte-associated diseases, Th1 lymphocyte-associated diseases, and activated B lymphocyte-associated diseases.

[0907] 53B. A method for preparing a camptothecin conjugate of any one of embodiments 1B-26B, said method comprising reacting an antibody having free thiol with a camptothecin-linker compound of any one of embodiments 27B-43B.

[0908] 54B. A kit comprising a camptothecin conjugate of any one of embodiments 1B-26B.

[0909] 55B. The kit according to implementation plan 54B further contains additional therapeutic agents.

[0910] 1C. A method having the formula L-(QD) p The camptothecin conjugate or its salt, wherein L is a ligand unit derived from a target agent, particularly from an antibody that selectively binds to cancer cell antigens; the subscript p is an integer from 1 to 16; and Q is a linker unit having a formula selected from: -ZA-, -ZA-RL-, -ZA-RL-Y-, -ZAS * -RL-、-ZAS * -RL-Y-、-ZAS * -W-、-ZAS * -W-RL-、-ZAB(S * )-RL-、-ZAB(S * -W- and -ZAB(S) * -W-RL- and -ZAB(S * )-RL-Y-, where

[0911] Z represents an extended sub-unit; A represents a key or connector unit; B represents a parallel connector unit; S * RL is a dispensing agent; W is a releasable linker; Y is a spacer unit; and D is a pharmaceutical unit selected from the following CPT1, CPT2, CPT3, CPT4, CPT5, CPT6, and CPT7:

[0912]

[0913]

[0914] Where R B The derivative is selected from H, C1-C8 alkyl, C1-C8 haloalkyl, C3-C8 cycloalkyl, (C3-C8 cycloalkyl)-C1-C4 alkyl, phenyl, and phenyl-C 1- Members of C4 alkyl groups; RC Selected from C1-C6 alkyl and C 3- Members of C6 cycloalkyl groups; R F and R F’ Each is independently selected from -H, C1-C8 alkyl, C1-C8 hydroxyalkyl, C1-C8 aminoalkyl, (C1-C4 alkylamino)-C1-C8 alkyl, N,N-(C1-C4 hydroxyalkyl)(C1-C4 alkyl)amino-C1-C8 alkyl, N,N-bis(C1-C4 alkyl)amino-C 1- C8 alkyl, N-C1-C4 hydroxyalkyl-C1-C8 aminoalkyl, C1-C8 alkyl C(O)-, C1-C8 hydroxyalkyl-C(O)-, C1-C8 aminoalkyl-C(O)-, C3-C 10 cycloalkyl, (C3-C 10 Cycloalkyl)-C1-C4 alkyl, C3-C 10 Heterocyclic alkyl groups, (C3-C 10 Members of heterocyclic alkyl (C1-C4 alkyl), phenyl, phenyl-C1-C4 alkyl, diphenyl-C1-C4 alkyl, heteroaryl and heteroaryl-C1-C4 alkyl; or R F and R F’ It combines with the nitrogen atoms to form a 5-, 6-, or 7-membered ring having 0 to 3 substituents selected from halogens, C1-C4 alkyl groups, -OH, -OC1-C4 alkyl groups, -NH2, and -NHC. 1- C4 alkyl and -N(C1-C4 alkyl)2; and wherein R B R C R F and R F’ The cycloalkyl, heterocycloalkyl, phenyl, and heteroaryl moieties are composed of 0 to 3 elements selected from halogens, C 1- C4 alkyl, -OH, -OC1-C4 alkyl, -NH2, -NHC 1- C4 alkyl and -N(C 1- Camptothecin compounds in which D is any one of 13a-13c of Table H, 14a-14z of Table I, and 18a-18r of Table J;

[0915] Where Q is -ZA-RL-, -ZA-RL-Y-, -ZAS * -RL-、-ZAB(S * -RL-、-ZAS * -RL-Y- or -ZAB(S) *When Q is -RL-Y-, the covalent bonding site of D is a heteroatom of any of the hydroxyl or amino substituents in any of CPT1, CPT2, CPT3, CPT4, CPT5, CPT6 or CPT7, or any of the hydroxyl or amino substituents in any of Table H 13a-13c, Table I 14a-14z and Table J 18a-18r, or where Q is -ZA-, -ZAS- * -W- or -ZAB(S * -W-, or when Q is -ZAS * -RL-、-ZAB(S * -RL-、-ZAS * -W-RL- or -ZAB(S * When RL is a releasable unit other than a glucuronide unit, the covalent linker of D is the oxygen atom of the hydroxyl substituent in the lactone ring of any one of CPT1, CPT2, CPT3, CPT4, CPT5, CPT6 or CPT7, or 13a-13c of Table H, 14a-14z of Table I, and 18a-18r of Table J; and

[0916] The condition is that when the covalent linkage site is the nitrogen atom of the amino substituent in CPT6, R F and R F’ At least one of them is -H, and the condition is that when D is a CPT1 having nitrogen atoms covalently linked through its amino substituents, -ZA-RL-, -ZA-RL-Y-, -ZAS * -RL-、-ZAB(S * -RL-、-ZAS * -RL-Y- and -ZAB(S * The -ZA- of )-RL-Y- is not the succinimide-hexanoyl-β-alanyl moiety of the succinimide ring in hydrolytic form.

[0917] 2C. The camptothecin conjugate according to embodiment 1C, wherein Q is a linker unit having the formula selected from –ZA-RL-, -ZA-RL-Y-, -ZAS*-RL-, -ZAB(S*)-RL-, -ZAS*-RL-Y- and –ZAB(S*)-RL-Y-, wherein A is a linker unit and RL is a glucuronic acid unit.

[0918] 3C. The camptothecin conjugate according to embodiment 2C, wherein the covalent linking point of D is the oxygen atom of the hydroxyl substituent on the lactone ring of any one of CPT1-CPT7.

[0919] 4C. The camptothecin conjugate according to embodiment 2C, wherein D is CPT1, CPT4, CPT6, or CPT7, wherein the covalent link with CPT1 is through the nitrogen atom of its amine functional group, provided that -ZA- is not an optional succinimide-hexanoyl-β-alanyl group with a hydrolyzed succinimide ring as the succinamide moiety.

[0920] The covalent linker of CPT4 is through the nitrogen atom of its amine functional group, and the covalent linker of CRF6 is through the nitrogen atom of its amine functional group, provided that R... F and R F’ At least one of them is -H, and the covalent connection point with CPT7 is through an oxygen atom, one of its primary hydroxyl functional groups.

[0921] 5C. Camptothecin conjugates according to embodiments 2C, 3C, or 4C, wherein the glucuronic acid unit has the following formula:

[0922]

[0923] Where Su is the hexose form of the monosaccharide; O' represents the oxygen atom of the glycosidic bond that can be cleaved by glycosidases; the wavy line marked with a single asterisk (*) indicates that R... F and R F’ At least one of these is a site covalently linked to the nitrogen atom of an amino substituent on CPT1, CPT4, or CPT6 of CPT1, or a site covalently linked to a spacer subunit (Y), or a site covalently linked to the oxygen atom of a hydroxyl substituent in the lactone ring of any one of CPT1-CPT7; and a wavy line marked with a double asterisk (**) indicates a site covalently linked to the remainder of Q.

[0924] In particular, the glucuronide unit has the following formula:

[0925]

[0926] 6C. The camptothecin conjugate according to embodiment 5C, wherein Q is a connecting subunit having the formula -ZA-RL-Y-, -ZAS*-RL-Y-, or -ZAB(S*)-RL-Y-; and the spacer subunit (Y) has the following formula:

[0927]

[0928] Where EWG is an electron-withdrawing group; O* represents the oxygen atom from the hydroxyl substituent of D; the wavy line adjacent to the nitrogen atom indicates the site covalently bonded to the carbonyl carbon atom of the glucuronide unit; and the wavy line adjacent to O* indicates the site covalently bonded to the rest of D, or

[0929] The spacer subunit (Y) has the following formula:

[0930]

[0931] When D is one of R F and R F’ Each of them is a -H CPT1, CPT4, or CPT6, and EWG is an electron-withdrawing group; the wavy line adjacent to the nitrogen atom indicates the site covalently linked to the carbonyl carbon atom of the glucuronide unit; and the wavy line adjacent to the carbonyl carbon atom indicates the site covalently linked to the nitrogen atom of the amino substituent of CPT1, CPT4, or CPT6.

[0932] 7C. The camptothecin conjugate according to embodiment 5C, wherein -ZA- is composed of a succinimide-alkanoyl moiety or a succinimide and triazole moiety, each optionally having a hydrolyzed succinimide ring as the succinic amide moiety, wherein the triazole moiety is optionally formed by the 1,3-dipolar cycloaddition of an azide substituent from a chemically modified target agent to an alkynyl moiety of a drug linker compound, wherein the target agent is a precursor of the conjugate ligand unit, or

[0933] -ZA- is composed of a succinic amide moiety derived from the mDPR portion of camptothecin-linker compounds, or of a succinimide-propionyl moiety optionally having a hydrolyzed succinimide ring.

[0934] The condition is that D has a nitrogen atom covalently linked through its amino substituent and -ZA- is composed of a succinimide group and a triazole group, which optionally has a succinimide ring in a hydrolyzed form as the succinamide moiety, or when D is CPT1, it is composed of a succinamide moiety that can be derived from the mDPR moiety, or the condition is that D has an oxygen atom covalently linked through a hydroxyl substituent on its lactone ring and when D is CPT1, -ZA- is composed of a succinimide-alkanoyl-β-alanyl group, which optionally has a succinimide ring in a hydrolyzed form as the succinamide moiety.

[0935] 8C. A camptothecin conjugate or a salt thereof according to embodiment 7C, wherein Q has the following formula:

[0936]

[0937] Wherein -ZA- is preferably a succinimide-alkanoyl-β-alanyl moiety having a hydrolyzed succinimide ring as the succinamide moiety, wherein the succinimide ring can be derived from the mDPR moiety of the camptothecin-linker compound; the wavy line marked with a single asterisk (*) indicates the oxygen atom of the hydroxyl functional group of the lactone ring that substitutes for any one of CPT1-CPT7, or the oxygen atom of the R in the lactone ring. Fand R F’ It is a nitrogen atom of an amine functional group of -H CPT1, CPT4, or CPT6, or a site covalently connected to a spacer subunit; and a wavy line marked with a three-star symbol (***) indicates a point covalently connected to a sulfur atom of L, or

[0938] Q has the following formula:

[0939]

[0940] When Q is -ZAS*-RL, a succinimide ring in hydrolyzed form is selected as the succinamide moiety, wherein the subscript n is an integer from 1 to 50, preferably 4; a wavy line marked with a single asterisk (*) indicates a heteroatom of any of the hydroxyl or amine functional groups of CPT1-CPT7 or a site covalently connected to the spacer subunit (Y); and a wavy line marked with a triplet (***) indicates a point covalently connected to the sulfur atom of L.

[0941] 9C. The camptothecin conjugate or its salt according to embodiment 6C, wherein -QD has the following structure:

[0942]

[0943] It optionally includes a succinimide ring in a hydrolyzed form as the succinamide moiety, where the wavy line indicates the site where the succinimide ring is covalently linked to the sulfur atom of the ligand unit, or

[0944] -QD has the following structure:

[0945]

[0946] Or its salts, or -QD, have the following structures:

[0947]

[0948] Or its salt, wherein the wavy line represents the site where the succinimide ring is covalently linked to the sulfur atom of the ligand unit, wherein the succinimide ring is the succinic amide moiety in hydrolytic form.

[0949] 10C. A camptothecin conjugate according to embodiment 1C, wherein Q is a connecting subunit having the form selected from –ZA-, -ZAS*-W- and –ZAB(S*)-W-, wherein A is a joint unit, or Q is a unit having the form selected from -ZA-RL-, -ZAS... * Connector units of the formulas -RL-, -ZAB(S*)-RL-, -ZAS*-W-RL-, and -ZAB(S*)-W-RL-, where A is a connector unit and RL is a releasable connector other than a glucuronide unit.

[0950] 11C. A camptothecin conjugate according to embodiment 10C, wherein Q is a connecting subunit having the formula selected from –ZA-RL-, -ZAS*-RL- and –ZAS*-W-RL-, wherein RL has the following formula:

[0951]

[0952] The wavy lines marked with double asterisks (**) indicate sites covalently connected to D; and the wavy lines marked with single asterisks (*) indicate sites covalently connected to A, S*, or W.

[0953] 12C. Camptothecin conjugates according to embodiment 10C or 11C, wherein -QD has the formula -ZAS*-W-RL-D, where D is the formula where R is the base of the conjugate. F and R F’ Each of them is a -H CPT1, CPT4, or CPT6, each having a nitrogen atom covalently linked to an amine functional group; and W is an amino acid unit selected from N-methyl-glycine (sarcosine), N-methyl-alanine, N-methyl-β-alanine, valine, N-methyl-valine, or D is a CPT1-CPT7 having an oxygen atom covalently linked to a hydroxyl substituent on the lactone ring; and W is an amino acid unit selected from glutamic acid or lysine.

[0954] 13C. The camptothecin conjugate according to embodiment 12C, wherein -ZA- is composed of a succinimide-alkanoyl moiety or a succinimide and triazole moiety, each optionally having a succinimide ring in a hydrolyzed form as the succinamide moiety, or a succinamide moiety derivable from the mDPR of the camptothecin-linker compound, or wherein -ZA- has the following formula:

[0955]

[0956] The succinimide ring in hydrolyzed form is optionally used as the succinamide moiety, wherein the wavy line marked with a double asterisk (**) indicates the site covalently linked to S*; and the wavy line marked with a triple star (***) indicates the point covalently linked to the sulfur atom of L.

[0957] 14C. A camptothecin conjugate according to embodiment 11C, wherein Q is a connecting subunit having a formula selected from -ZAS*-RL- and –ZAS*-W-RL-, wherein S* has the following formula:

[0958]

[0959] Where the subscript n is an integer from 2 to 36, the wavy line adjacent to the nitrogen atom indicates the site covalently bonded to the carbonyl carbon atom of A, and the wavy line adjacent to the carbonyl carbon atom indicates the site covalently bonded to the nitrogen atom of the amine functional group of RL in -ZAS*-RL- or W in –ZAS*-W-RL-, especially the –Z A- of Q in any formula has the following:

[0960]

[0961] The wavy lines marked with double asterisks (**) indicate sites covalently bonded to the nitrogen atom of the amine functional group of S*; and the wavy lines marked with triple asterisks (***) indicate sites covalently bonded to the sulfur atom of L.

[0962] 15C. Camptothecin conjugate according to embodiment 11C, wherein Q is a connecting subunit having the formula –ZAS*-W- or –ZASW-RL-, wherein –ZAS*-W- in either formula has the following expression:

[0963]

[0964] The succinimide ring in hydrolyzed form is optionally used as the succinamide moiety, wherein the subscript n is an integer from 2 to 10, preferably an integer from 2 to 4; wherein the wavy line marked with a double asterisk (**) indicates the site covalently connected to D or RL; and the wavy line marked with a triple star (***) indicates the point covalently connected to the sulfur atom of L.

[0965] 16C. A camptothecin conjugate or a salt thereof according to embodiment 10C, wherein –QD has the following structure:

[0966]

[0967]

[0968] The wavy line represents the point where the succinimide ring, which is optionally hydrolyzed as the succinamide moiety, is covalently bonded to the sulfur atom of the ligand unit.

[0969] 17C. Camptothecin-linker compounds having a formula selected from the following:

[0970] (i)Z'-A-RL-D;

[0971] (ii)Z'-A-RL-YD;

[0972] (iii)Z'-AS * -RL-D;

[0973] (iv)Z'-AS * -RL-YD;

[0974] (v)Z'-AB(S * )-RL-D;

[0975] (vi)Z'-AB(S * )-RL-YD;

[0976] (vii)Z'-AD

[0977] (viii)Z'-AS*-WD

[0978] (ix)Z'-AB(S*)-WD

[0979] (x)Z'-AS*-W-RL-D; and

[0980] (xi)Z'-AB(S*)-W-RL-D

[0981] Where Z' is the precursor of the extended subunit; A is the key or connector unit; B is the parallel connector unit; S * is a distributing agent; RL is a releasable linker; Y is a spacer subunit; and D is a camptothecin compound selected from the following CPT1, CPT2, CPT3, CPT4, CPT5, CPT6 and CPT7:

[0982]

[0983] R B The derivative is selected from H, C1-C8 alkyl, C1-C8 haloalkyl, C3-C8 cycloalkyl, (C3-C8 cycloalkyl)-C1-C4 alkyl, phenyl, and phenyl-C 1- C4 alkyl moiety; R C Selected from C1-C6 alkyl and C 3- C6 cycloalkyl moiety; R F and R F’ Each is independently selected from -H, C1-C8 alkyl, C1-C8 hydroxyalkyl, C1-C8 aminoalkyl, (C1-C4 alkylamino)-C1-C8 alkyl, N,N-(C1-C4 hydroxyalkyl)(C1-C4 alkyl)amino-C1-C8 alkyl, N,N-bis(C1-C4 alkyl)amino-C 1- C8 alkyl, N-C1-C4 hydroxyalkyl-C1-C8 aminoalkyl, C1-C8 alkyl C(O)-, C1-C8 hydroxyalkyl-C(O)-, C1-C8 aminoalkyl-C(O)-, C3-C 10 cycloalkyl, (C3-C 10 Cycloalkyl)-C1-C4 alkyl, C3-C 10 Heterocyclic alkyl groups, (C3-C 10Members of heterocyclic alkyl (C1-C4 alkyl), phenyl, phenyl-C1-C4 alkyl, diphenyl-C1-C4 alkyl, heteroaryl and heteroaryl-C1-C4 alkyl; or R F and R F’ It combines with the nitrogen atoms to form a 5-, 6-, or 7-membered ring having 0 to 3 substituents selected from halogens, C1-C4 alkyl groups, -OH, -OC1-C4 alkyl groups, -NH2, and -NHC. 1- C4 alkyl and -N(C1-C4 alkyl)2; and wherein R B R C R F and R F’ The cycloalkyl, heterocycloalkyl, phenyl, and heteroaryl moieties are composed of 0 to 3 elements selected from halogens, C 1- C4 alkyl, -OH, -OC1-C4 alkyl, -NH2, -NHC 1- C4 alkyl and -N(C 1- Substituents of C4 alkyl group 2; and

[0984] Wherein, when the camptothecin-linker compound is of formula (i), (ii), (iii), (iv), (v), or (vi), the covalent linking point of D is a heteroatom of any one of the hydroxyl or amino substituents of any one of CPT1-CPT7; or when the camptothecin-linker compound is of formula (vii), (viii), or (ix), or when the camptothecin-linker compound is of formula (iii), (iv), (x), or (xi), where RL is a releasable unit other than the glucuronide unit, the covalent linking point of D is an oxygen atom of the hydroxyl substituent on the lactone ring of any one of CPT1-CPT7; and

[0985] The condition is that when the covalent linkage site is the nitrogen atom of the amino substituent in CPT6, R F and R F’ At least one of them is -H, and the condition is that when D is a CPT1 having a nitrogen atom covalently linked by its amino substituent, the Z'-A- of the camptothecin-linker compounds of formulas (i), (ii), (iii), (iv), (v) and (vi) is not a maleimide-hexanoyl-β-alanyl moiety.

[0986] 18C. The camptothecin-linker compound according to embodiment 17C has a formula selected from formulas (i), (ii), (iii), (iv), (v), and (vi), wherein A is a linker unit and RL is a glucuronide unit, and particularly has the following formula:

[0987]

[0988] A wavy line marked with a single asterisk (*) indicates a site covalently connected to D or to the spacer subunit (Y); and a wavy line marked with a double asterisk (**) indicates a site covalently connected to A, B, or S*.

[0989] 19C. Camptothecin-linker compound according to embodiment 18C, wherein the covalent linking point of D is the oxygen atom of a hydroxyl substituent on the lactone ring of any one of CPT1-CPT7.

[0990] 20C. Camptothecin-linker compounds according to embodiment 18C, wherein D is CPT1, CPT4, or CPT6, wherein the linker of CPT1 is through the nitrogen atom of its amine functional group, provided that Z'-A- is not maleimide-hexanoyl-β-alanyl; the linker of CPT4 is through the nitrogen atom of its amine functional group; and the linker of CPT6 is through the nitrogen atom of its amine functional group, provided that R... F and R F’ At least one of them is -H.

[0991] 21C. The camptothecin-linker compound according to embodiment 17C, having formulas (iii), (iv), (v) and (vi), wherein S * It is a PEG group.

[0992] 22C. The camptothecin-linker compound according to embodiment 18C, having formula (ii), formula (iv) or formula (vi), wherein D is any one of CPT1-CPT7; and the spacer subunit (Y) has the following formula:

[0993]

[0994] Where EWG is an electron-withdrawing group; O* represents the oxygen atom from the hydroxyl functional group of D; the wavy line adjacent to the nitrogen atom indicates the site covalently bonded to the carbonyl carbon atom of the glucuronide unit; and the wavy line adjacent to O* indicates the site covalently bonded to the rest of D, or D is selected from R. F and R F’ Each of them is -H CPT1, CPT4 and CPT6; and the spacer subunit (Y) has the following formula:

[0995]

[0996] Where EWG is an electron-withdrawing group; the wavy line adjacent to the nitrogen atom indicates the site covalently bonded to the carbonyl carbon atom of the glucuronide unit; and the wavy line adjacent to the carbonyl carbon atom indicates the site covalently bonded to the R atom. F and R F’ Each of these is a site covalently linked to the nitrogen atom of an amine functional group of CPT1, CPT4, or CPT6 with a -H atom.

[0997] 23C. A camptothecin-linker compound according to any one of embodiments 17C-22C, wherein A is composed of an alkynyl moiety capable of 1,3-dipolar cycloaddition with an azide substituent from a chemically modified target agent that is a precursor of a ligand unit of the camptothecin conjugate, thereby providing a conjugate having a linker unit composed of a triazole moiety.

[0998] 23C. A camptothecin-linker compound according to any one of embodiments 17C-22C, wherein Z'-A- is composed of a maleimide-alkanoyl moiety or mDPR, the basic nitrogen atom of which is optionally protonated or protected by an acid-labile protecting matrix, provided that when D is a CPT1 having a nitrogen atom covalently linked through its amino substituent, Z'-A- is composed of mDPR, particularly Z'-A- is composed of mDPR or a maleimide-alkanoyl-β-alanyl moiety, provided that when D is a CPT1, D has a covalently linked oxygen atom to a hydroxyl substituent on its lactone ring.

[0999] 24C. The camptothecin-linker compound according to embodiment 17C, having formula (vii), formula (viii) or formula (ix), wherein A is a linker unit, or having formula (i), formula (iii), formula (x) or formula (xi), wherein A is a linker unit and RL is a releasable linker other than a glucuronide unit.

[1000] 25C. The camptothecin-linker compound according to embodiment 24C, having formula (i), formula (iii), or formula (x), wherein RL has the following formula:

[1001]

[1002] The wavy lines marked with double asterisks (**) indicate sites covalently connected to D; and the wavy lines marked with single asterisks (*) indicate sites covalently connected to A, S*, or W.

[1003] 26C. The camptothecin-linker compound according to embodiment 25C has the formula (x), wherein W is an amino acid unit selected from N-methyl-glycine (sarcosine), N-methyl-alanine, N-methyl-β-alanine, valine and N-methyl-valine.

[1004] 27C. Camptothecin-linker compounds according to embodiments 24C, 25C or 26C, wherein Z'-A- is composed of a maleimide-alkanoyl moiety or mDPR, and its basic nitrogen atom is optionally protonated or protected by an acid-instable protecting matrix.

[1005] 28C. Camptothecin-linker compounds according to embodiments 24C, 25C or 26C, having formula (iii) or formula (x), wherein Z'-A- has a formula selected from:

[1006]

[1007] The wavy lines marked with double asterisks (**) indicate sites covalently connected to S*.

[1008] 29C. Camptothecin-linker compounds according to embodiments 24C, 25C or 26C, having formula (iii) or formula (x), wherein S* has the following formula:

[1009]

[1010] The subscript n is an integer between 2 and 36.

[1011] 30C. Camptothecin-linker compounds of formula (viii) or formula (x) according to embodiment 24C or 25C, wherein Z'-AS*-W- has the following formula:

[1012]

[1013] The subscript n is an integer from 2 to 10, preferably an integer from 2 to 4; the wavy line marked with a double asterisk (**) indicates a site covalently connected to D or RL.

[1014] 31C. The camptothecin-linker compound or its salt according to embodiment 17C has the following structure:

[1015]

[1016]

[1017]

[1018] 32C. Use of camptothecin conjugates in the preparation of medicaments for treating cancer in a subject, wherein the camptothecin conjugate has the formula of embodiment 1C, particularly wherein the cancer is selected from lymphoma, leukemia and solid tumors, preferably lymphoma or leukemia.

[1019] 33C. A pharmaceutically acceptable composition comprising the camptothecin conjugate of embodiment 1C and at least one pharmaceutically acceptable excipient.

[1020] 34C. A composition for treating cancer in a subject who requires it, wherein the composition comprises an effective amount of the camptothecin conjugate of embodiment 1C, wherein the cancer is selected from lymphoma, leukemia and solid tumors, preferably lymphoma or leukemia.

[1021] 35C. A method for preparing the camptothecin conjugate of embodiment 1, the method comprising the step of contacting a targeting agent having a functional group reactive to Z' of the camptothecin-linker compound of claim 17, thereby forming a covalent bond between the ligand unit and the extended subunit (Z) of the camptothecin conjugate, whose structures respectively correspond to the targeting agent and Z', particularly

[1022] The targeting agent is an antibody having at least one cysteine ​​residue, wherein the reactive functional group is a thiol and Z' is composed of a maleimide moiety, or the targeting agent is an antibody modified to have an azide-containing residue as a reactive functional group and Z' is composed of an alkyne functional group, wherein the azide and alkyne functional groups are capable of undergoing a 1,3-dipolar cycloaddition reaction to form a triazole ring system.

[1023] Detailed Implementation Plan

[1024] Materials and methods

[1025] Unless otherwise stated, the following materials and methods are applicable to the synthetic procedures described in this section. All commercially available anhydrous solvents can be used without further purification. Starting materials, reagents, and solvents were purchased from commercial suppliers (SigmaAldrich and Fischer). Biotage Isolera One was used. TM The rapid purification system (Charlotte, NC) purifies the product using rapid column chromatography. Using the UPLC method shown in Table AF, in conjunction with Waters Acquity... TM UPLC-MS was performed on a Waters single quadrupole mass spectrometer with a UPLC system interface. Preparative HPLC was performed on a Waters 2454 binary gradient module solvent delivery system equipped with a Wasters 2998 PDA detector. A Phenomenex Max-RP 4μm Synergid lens of suitable diameter was used. TM Purify the product using a 250 mm reversed-phase column, eluting with 0.05% trifluoroacetic acid / water and 0.05% trifluoroacetic acid / acetonitrile, unless otherwise specified.

[1026] Table A: Column - Waters Acuity UPLC BEH C18 2.1 x 50 mm, 1.7 μm, reversed-phase column, solvent A - 0.1% formic acid aqueous solution, solvent B - acetonitrile containing 0.1% formic acid (method A).

[1027] Time (min) Flow rate (mL / min) A％ B％ gradient Start 0.5 97 3 1.70 0.5 40 60 linear 2.00 0.5 5 95 linear 2.50 0.5 5 95 linear 2.80 0.5 97 3 linear 3.00 0.5 97 3 linear

[1028] Table B: Column - Waters Acuity UPLC BEH C18 2.1 x 50 mm, 1.7 μm, reversed-phase column, solvent A - 0.1% formic acid aqueous solution, solvent B - acetonitrile containing 0.1% formic acid (method B).

[1029] Time (min) Flow rate (mL / min) A％ B％ gradient Start 0.6 97 3 1.50 0.6 5 95 linear 2.40 0.6 5 95 linear 2.50 0.6 97 3 linear 2.80 0.6 97 3 linear

[1030] Table C: Column - Kinetex F5 1.7μm 2.1 x 50 mm, reversed-phase column, solvent A - 0.1% formic acid aqueous solution, solvent B - acetonitrile containing 0.1% formic acid (method C).

[1031] Time (min) Flow rate (mL / min) A％ B％ gradient Start 0.5 97 3 2.50 0.5 40 95 linear 3.50 0.5 5 95 linear 3.75 0.5 97 3 linear 4.00 0.5 97 3 linear

[1032] Table D: Column - Waters CORTECS C18 1.6μm, 2.1 x 50mm, reversed-phase column, solvent A - 0.1% formic acid aqueous solution, solvent B - acetonitrile containing 0.1% formic acid (method D).

[1033] Time (min) Flow rate (mL / min) A％ B％ gradient Start 0.6 97 3 1.70 0.6 40 60 linear 2.00 0.6 5 95 linear 2.50 0.6 5 95 linear 2.80 0.6 97 3 linear 3.00 0.6 97 3 linear

[1034] Table E: Column - Waters CORTECS C18 1.6μm, 2.1 x 50mm, reversed-phase column, solvent A - 0.1% formic acid aqueous solution, solvent B - acetonitrile containing 0.1% formic acid (Method E).

[1035] Time (min) Flow rate (mL / min) A％ B％ gradient Start 0.6 97 3 1.50 0.6 5 95 linear 2.40 0.6 5 95 linear 2.50 0.6 97 3 linear 2.80 0.6 97 3 linear

[1036] Table F: Column - Waters CORTECS C8 1.6μm, 2.1 x 50mm, reversed-phase column, solvent A - 0.1% formic acid aqueous solution, solvent B - acetonitrile containing 0.1% formic acid (method F).

[1037]

[1038]

[1039] Table G: List of Abbreviations

[1040]

[1041] Preparation of camptothecin compounds

[1042] The camptothecin compounds provided in the following examples can be used to prepare camptothecin-linker compounds and camptothecin conjugates as described herein.

[1043] Example 1

[1044]

[1045] SN-38 (compound 1, 160.0 mg, 0.4077 mmol), purchased from MedChemExpress, was suspended in anhydrous DCM (2 mL). DIPEA (0.22 mL, 1.3 mmol) was added, followed by TBSCl (154 mg, 1.02 mmol). The reaction was stirred for 30 min until compound 1 became soluble and complete conversion was observed by UPLC-MS. The reaction was quenched with MeOH, filtered through a silica gel stopper, and concentrated under vacuum. The resulting colorless oil was ground with Hex. The product precipitated from the solution. The precipitate was collected by filtration and washed with Hex to give compound 2 (TBS-SN-38) (200 mg, 0.395 mmol, 97%) as a grayish-white solid. LC-MS (Method B): t R = 1.86 min; MS (m / z) [M+H] + C 28 H 35 The calculated value of N2O5Si is 507.23, and the measured value is 506.96.

[1046] Example 2

[1047]

[1048] Compound 3 was synthesized according to the procedure described in Bioconjugate Chem. 2009, 20, 1242–1250. Compound 3 (50 mg, 0.108 mmol) was dissolved in DCM (1 mL). DMAP (13 mg, 0.11 mmol) was added to the reaction mixture, followed by Boc2O (24 mg, 0.11 mmol). The reaction was stirred for 5 minutes, at which point complete conversion to the desired product was observed. The protected product was purified by column chromatography using 10 G Biotage Ultra, 0–5% MeOH / DCM. The fraction containing the desired product was concentrated under vacuum to give compound 4 (49 mg, 0.087 mmol, 80%) as a yellow solid. LC-MS (Method A): t R = 2.24 min; MS(m / z) [M+H] + C 30 H 34 The calculated value of N3O8 is 564.23, and the measured value is 564.10.

[1049] Compound 4 (49 mg, 0.087 mmol) was dissolved in anhydrous DCM (2 mL). DMAP (37 mg, 0.304 mmol) was added and the reaction was cooled to 0 °C. Triphosgene (12 mg, 0.039 mmol) dissolved in DCM at a concentration of 10 mg / mL was added dropwise over 15 minutes. A 2 μL aliquot was quenched in 98 μL of MeOH diluent and injected onto UPLC-MS. Complete conversion to the MeOH adduct was observed by UPLC-MS. This reaction mixture (compound 5) can be used directly in coupling steps with a suitable linker. LC-MS (Method A): t R = 2.09 min; MS (m / z) [M+H] + C 32 H 36 N3O 10 Calculated value: 622.24; measured value: 622.02.

[1050] Example 3

[1051]

[1052] Compound 6 (150 mg, 0.334 mmol) was synthesized according to the method described in Bioconjugate Chem. (2009) 20:1242–1250 and dissolved in anhydrous DCM (2 mL). DMAP (143 mg, 1.17 mmol) was added. Triphosgene (45 mg, 0.15 mmol) (50 mg / mL) dissolved in anhydrous DCM was added dropwise over 5 minutes. The reaction was stirred at room temperature for 30 minutes. A 2 μL aliquot of the reaction mixture was quenched in 98 μL of MeOH diluent. Almost complete conversion to MeOH carbonate was observed, indicating the formation of chloroformate. Compound 7 thus obtained can be used directly in the coupling step with a suitable linker without further purification. LC-MS (Method A): t R = 1.55 min; MS (m / z) [M+H] + C 27 H 27 The calculated value of N2O8 is 507.18, and the measured value is 507.06.

[1053] Example 4

[1054]

[1055] 6-Amino-3,4-(methylenedioxy)acetophenone (8, 5.00 g, 27.9 mmol), obtained from TCI Research Chemicals (Cat. No. A1356), was dissolved in DCM (100 mL). The reaction was cooled to 0 °C and DIPEA (7.29 mL, 41.9 mmol) was added, followed by the slow addition of acetyl chloride (2.49 mL, 34.9 mL). The reaction was heated to room temperature and stirred for 30 min. Complete conversion was observed by UPLC-MS. The reaction was quenched with MeOH (5 mL) and concentrated under vacuum to give compound 9 as a white solid, which was used in the next step without further purification. LC-MS (Method A): t R =1.37; MS(m / z)[M+H] + C 11 H 12 The calculated value of NO4 is 222.08, and the measured value is 222.11.

[1056] Compound 9 (27.9 mmol) was dissolved in AcOH (100 mL). 33% w / w HBr / AcOH (9.78 mL, 55.8 mmol) was slowly added. Bromine (1.44 mL, 27.9 mmol) was added dropwise over 15 minutes. The reaction was stirred for 30 minutes, at which point the conversion to the desired product was observed. The reaction mixture was poured over ice water, and the precipitate was collected by filtration and washed with water. The filtrate was dried to give a yellow powder, which was a mixture of the desired product compound 10 with the starting material and dibromination product impurities, used in the next step without further purification (7.2 g, 24 mmol, 86%). LC-MS (Method A): t R = 1.58 min; MS (m / z) [M+H] + C 11 H 11 The calculated value of BrNO4 is 299.99, and the measured value is 299.90.

[1057] Compound 10 (7.2 g, 24 mmol) was dissolved in EtOH (100 mL). Concentrated HBr (5 mL) was added, and the reaction was heated to reflux and maintained for 60 minutes. Almost complete conversion to the deprotected product was observed. The reaction was concentrated under vacuum and diluted with DCM (200 mL) and H2O (200 mL). The aqueous phase was extracted with DCM (3 x 200 mL), and the collected organic phase was dried over MgSO4, filtered, and concentrated under vacuum. The crude product was purified by column chromatography with 0–10% MeOH / DCM. The fraction containing the desired product and minor impurities was concentrated to give compound 11 (4.05 g, 15.7 mmol, 65%) as a yellow powder. LC-MS (Method A): t R= 1.57 min; MS (m / z) [M+H] + The calculated value of C9H9BrNO3 is 257.98, and the measured value is 257.71.

[1058] Example 5

[1059]

[1060] Compound 11 (1.00 g, 3.87 mmol), p-TSA (667 mg, 3.87 mmol), and 4-ethyl-4-hydroxy-7,8-dihydro-1H-pyrano[3,4-f]indazine-3,6,10(4H)-trione (1.02 g, 3.87 mmol, obtained from Avra ​​Laboratories Pvt. Ltd.) were added to a flask to homogenize the solid, and then evaporated under nitrogen. The purified solid was then heated to 120 °C under high vacuum (1 mbar) for 60 min. The reaction was cooled to room temperature, and the crude product was precipitated with H2O, filtered, and washed with H2O. The precipitate was purified by column chromatography with 0–10% MeOH / DCM. The fraction containing the desired product was concentrated under vacuum to give compound 12 (989 mg, 2.04 mmol, 53%) as a brown solid. LC-MS (Method A): t R = 1.62 min (Universal method UPLC); MS (m / z) [M+H] + C 22 H 17 The calculated value of BrN2O6 is 485.03, and the measured value is 484.95.

[1061] Example 6

[1062]

[1063] Compound 12 (188 mg, 0.387 mmol) was dissolved in EtOH (5 mL). Hexamethylenetetramine (163 mg, 1.16 mmol) was added, and the reaction was stirred under reflux for 90 minutes. The reaction was cooled, and concentrated HCl aqueous solution (0.1 mL) was added. The reaction was concentrated, and purified by preparative-HPLC. The fraction containing the desired product was lyophilized to give compound 13 (109 mg, 0.259 mmol, 67%) as a white solid. LC-MS (Method A): t R =0.89; MS(m / z)[M+H] + C 22 H 20 The calculated value of N3O6 is 422.14, and the measured value is 422.16.

[1064] Table H: Camptothecin compound (7-MAD-MDCPT) prepared from compound 13

[1065]

[1066] The camptothecin compounds in Table H are exemplary compounds of formula W-CPT, which are incorporated into camptothecin conjugates of formula -ZAS*-WD or -ZAB(S*)-WD, or incorporated into pharmaceutical linker compounds of formula Z'-AS*-WD or Z'-AB(S*)-WD by covalent linking to the oxygen or nitrogen atom of the primary hydroxyl or amine functional group, respectively.

[1067] Example 7

[1068]

[1069] Compound 12 (10.0 mg, 20.6 μmol) from Example 4 was dissolved in anhydrous DMF (0.25 mL). Methylamine (2 M in THF, 0.031 mL, 62 μmol) was added. The reaction was stirred for 30 min and then quenched with AcOH (20 μL). The reaction was purified by preparative HPLC. The fraction containing the desired product (14) was lyophilized to give a yellow solid (3.27 mg, 7.51 μmol, 36%). LC-MS (Method D): t R = 1.57 min; MS (m / z) [M+H] + The calculated value of C9H9BrNO3 is 257.98, and the measured value is 257.71. R = 0.93 min (Method A). MS(m / z)[M+H] + C 23 H 22 The calculated value of N3O6 is 436.15, and the measured value is 435.78.

[1070] Table I: Other camptothecin compounds prepared according to Example 7

[1071]

[1072]

[1073]

[1074]

[1075]

[1076]

[1077] *LC-MS Method A

[1078] Example 8

[1079]

[1080] 6-Nitro-1,3-benzodioxono-5-carboxynitrile (compound 15, 2.00 g, 10.4 mmol) was prepared as described in Heterocycles, (2007) 71:39-48, and then dissolved in EtOH (50 mL). The reaction was carried out under a nitrogen atmosphere. Pd / C (2.22 g, 10% w / w, 2.08 mmol) was added to the reaction, and the reaction was carried out under a hydrogen atmosphere. The reaction was stirred for 2 hours. The reaction was filtered through a diatomaceous earth (Celite) bed and then washed with MeOH. The eluent was concentrated under vacuum and purified by rapid chromatography with 0-10% DCM / MeOH. The fraction containing the desired product was concentrated to give compound 16 (1.46 g, 9.00 mmol, 87%) as a red solid. LC-MS (Method D): t R = 1.14 min; MS (m / z) [M+H] + The calculated value of C8H7N2O2 is 163.05, and the measured value is 162.37.

[1081] Example 9

[1082]

[1083] 6-Amino-1,3-benzodioxono-5-carboxynitrile (compound 16, 50 mg, 0.31 mmol) was dissolved in anhydrous THF (1 mL) under a nitrogen atmosphere. CuBr (1.5 mg, 0.010 mmol) was added, followed by 1 M 4-fluorophenyl magnesium bromide in THF (1.23 mL). The reaction was heated to 60 °C and held for 30 min, then cooled to room temperature. A 15% H₂SO₄ solution was slowly added to the solution, followed by stirring for 30 min. The reaction mixture was poured into saturated NaHCO₃ (50 mL) and extracted with EtOAc (3 x 50 mL). The organic matter was dried over MgSO₄, filtered, and concentrated under vacuum. The crude product was purified by column chromatography using 10 G Biotage Ultra, 0-10% EtOAc / Hex. The fraction containing the desired product was concentrated under vacuum to give compound 17 (46.2 mg, 0.178 mmol, 58%) as a red solid. LC-MS (Method D):t R = 1.81 min; MS (m / z) [M+H] + C 14 H 11 The calculated value of FNO3 is 260.07, and the measured value is 259.46.

[1084] Example 10

[1085]

[1086] Compound 17 (46.2 mg, 0.178 mmol), p-TSA (30.7 mg, 0.178 mmol), and 4-ethyl-4-hydroxy-7,8-dihydro-1H-pyrano[3,4-f]indazine-3,6,10(4H)-trione (46.9 mg, 0.178 mmol, obtained from Avra ​​Laboratories Pvt. Ltd.) were added to a scintillation vial to homogenize the solid. The solvent was concentrated under nitrogen. The purified solid was heated to 120 °C under high vacuum (1 mbar) for 60 min. The reaction was reconstituted in DCM (50 mL), washed with H2O, the organic phase was dried over MgSO4, filtered, and concentrated under vacuum. The crude product was purified by column chromatography using 10 G Biotage Ultra, 0–10% MeOH / DCM. Vacuum concentration of the fraction containing the desired product (18) yielded a red solid (32.9 mg, 0.0676 mmol, 38%). LC-MS (Method D): t R = 1.81 min; MS (m / z) [M+H] + C 27 H 20 The calculated value of FN2O6 is 487.13, and the measured value is 487.19.

[1087] Table J: Other camptothecin compounds prepared according to the methods of Examples 9 and 10

[1088]

[1089]

[1090]

[1091]

[1092] *LC-MS method A, except for 18q (method E) and 18r (method D)

[1093] Example 11

[1094]

[1095] SN-38 (compound 1, 76.0 mg, 0.19 mmol) obtained from MedChemExpress was dissolved in dichloromethane, followed by the addition of triethylamine (128 μL, 0.92 mmol) and DMAP (2.60 mg, 0.02 mmol). The mixture was cooled to 0 °C in an ice bath, and then acetyl chloride (15.9 μL, 0.22 mmol) was added dropwise. The reaction mixture was stirred at room temperature for 16 hours. The reaction mixture was diluted with dichloromethane and washed with saturated NH4Cl, water, and brine. The organic phase was then dried over MgSO4, filtered, concentrated, and purified on silica gel by Biotage rapid column chromatography (CH2Cl2 / MeOH 0-15%) to yield acetylated SN-38 (19). MS (m / z) calculated value 435.15 (M+H) + The measured value is 435.07.

[1096] Table K: Camptothecin compounds prepared as described in this article

[1097]

[1098]

[1099] Example 12

[1100]

[1101] Ecinotecan mesylate (compound 21a, 20.0 mg, 0.0376 mmol, obtained from MedChemExpressCat.No.:HY-13631A) was suspended in anhydrous DCM (1 mL). DIPEA (20.0 μL, 0.0146 mmol) was added, followed by acetoxyacetyl chloride (5.0 μL, 0.046 mmol). The reaction was stirred for 30 min, then quenched with MeOH and concentrated under vacuum. The reaction mixture was redissolved in MeOH (1 mL). LiOH (20 mg) was added. Complete deprotection with acetic acid was observed. AcOH was quenched. Purification was performed by preparative HPLC (10 mmol / L 95% MeCN in 0.05% TFA in H₂O). The fraction containing the desired product was concentrated under vacuum to give compound 21b (15.3 mg, 0.0310 mmol, 82%) as a yellow solid. LC-MS (Method A): t R = 1.46 min; MS (m / z) [M+H] + C 26 H 25 The calculated value of N3O6 is 494.17, and the measured value is 494.05.

[1102] Example 13

[1103]

[1104] Ecinotecan mesylate (compound 21a, 20.0 mg, 0.0376 mol) was dissolved in MeCN (1 mL) and NaHCO3 0.75 M H2O. Fmoc-OSu (19.0 mg, 0.0564 mmol) was added, and the reaction was stirred for 2 hours and 30 minutes. The reaction mixture was diluted with H2O (50 mL), the pH was adjusted to neutral, and the mixture was extracted with DCM (3 x 50 mL). The organic phase was dried over MgSO4, filtered, and concentrated under vacuum. The crude product was purified by preparative TLC using 0–5% MeOH in DCM. The band containing the desired product was scraped off, filtered, washed with 10% MeOH in DCM, and the eluent was concentrated under vacuum to give compound 22 (19.1 mg, 0.0290 mmol, 77%) as an orange solid. LC-MS (Method A): t R = 2.23 min; MS (m / z) [M+H] + C 39 H 33 The calculated value of FN3O6 is 658.24, and the measured value is 658.09.

[1105] Example 14

[1106]

[1107] Compound 2 (132 mg, 0.260 mmol) prepared according to Example 1 was dissolved in 2 mL of anhydrous DCM. DMAP (111 mg, 0.911 mmol) was added. Triphosgene (34.8 mg, 0.117 mmol) was dissolved in anhydrous DCM at a concentration of 50 mg / mL, and the solution was added dropwise to the stirred reaction solution over 5 minutes. 2 μL of the reaction solution was quenched in 98 μL of MeOH diluent. After 15 minutes, almost complete conversion to Me-carbonate was observed by UPLC-MS. The reaction mixture containing compound 24 was immediately used in the coupling reaction described herein.

[1108] Preparation of camptothecin drug linker compounds

[1109] Example 15

[1110]

[1111] Ecinotecan mesylate (compound 21a, 5.00 mg, 9.41 μmol) was dissolved in anhydrous DCM. DIPEA (5.0 μL, 28 μmol) was added, followed by compound 25 (17.2 mg, 18.8 μmol), previously described by Bioconjugate Chem. (2006) 17:831-840. The reaction was stirred at 40 °C for 3 h. The reaction was quenched with MeOH and concentrated under vacuum. The crude reaction mixture was used for the next step. LC-MS (Method A): t R = 2.32 min; MS (m / z) [M+H] + C 63 H 61 FN5O 19 Calculated value: 1210.39; measured value: 1210.08.

[1112] The crude compound 26 (9.41 μmol) from the previous step was dissolved in a solution of THF (1 mL) and 1 M LiOH in MeOH (1 mL). The reaction was stirred for 5 min, then H₂O was added and stirred for another 5 min. The reaction was quenched with AcOH (100 μL), concentrated under vacuum, and purified by preparative HPLC 21 mm 5-60-95% MeCN in 0.05% TFA in H₂O. The fraction containing the desired product was lyophilized to give compound 27 (1.1 mg, 1.3 μmol) as a yellow powder. LC-MS (Method A): t R = 1.29 min; MS (m / z) [M+H] + C 41 H 43 FN5O 14 Calculated value: 848.28; Measured value: 848.03.

[1113] Example 16

[1114]

[1115] Compound 27 (1.1 mg, 1.3 μmol) was dissolved in anhydrous DMF (0.5 mL). DIPEA (1 μL) was added, followed by N-succinimide-3-maleimide propionate (28, 0.63 mg, 2.4 μmol), purchased from TCI (CAS: 55750-62-4). The reaction was stirred for 5 min. Complete conversion was observed by UPLC-MS. The reaction was quenched with AcOH (10 μL) and purified by preparative HPLC 10 mm 5-60-95% MeCN in 0.05% TFA in H2O. The fraction containing the desired product was lyophilized to give compound 29 (1.21 mg, 1.21 μmol, 93%) as a yellow powder. LC-MS (Method A): tR = 1.52 min; MS (m / z) [M+H] + C 48 H 48 FN6O 17 The calculated value is 999.31, and the measured value is 999.07.

[1116] Example 17

[1117]

[1118] Compound 30 (210 mg, 0.234 mmol), prepared as described in Examples 21 and 22, was dissolved in standard DCM (3 mL). Paraformaldehyde (300–600 mg, xs) was added. The mixture was stirred vigorously, and TMSBr (0.1 mL) was added. The reaction mixture was stirred for 10 min, at which point complete conversion was observed by UPLC-MS. The reaction mixture was filtered through a syringe filter, washed with DCM (2 x 3 mL), and toluene (3 mL) was added to azeotropically bring the final mixture to a boil. The mixture was concentrated under vacuum to give a white solid. No further purification was required for the next step. The MeOH-quenched adduct was observed by UPLC-MS using MeOH diluent. LC-MS (Method A): t R = 2.19 min; MS (m / z) [M+Na] + C 44 H 51 N3NaO 18 The calculated value of S is 964.28, and the measured value is 965.17.

[1119] Example 18

[1120]

[1121] Prior to use, compound 20c (20 mg, 0.047 mmol), designated 7-BAD-MDCPT, was azeotropically reacted three times with toluene and dried under high vacuum. Crude compound 31 (231 mg, 0.234 mmol) from Example 16 was dissolved in anhydrous DCM, and 1,2,2,6,6-pentamethylpiperidine (PMP, 51.4 μL, 0.284 mmol) was added. Minimal hydrolysis of compound 31 in solution was observed by UPLC-MS after the addition of base. The solution of compound 31 was added directly to the drug reaction vessel and then heated to reflux. Compound 20c was only slightly soluble in DCM. The degree of reaction completion was monitored by UPLC-MS, which required reflux for 3 days. The reaction was then quenched with MeOH, concentrated under vacuum, and purified by DCM solution of 0-10% MeOH using an FCC Biotage 10G Ultra. The fraction containing the desired product (compound 32) was concentrated to give a yellow solid (50 mg, ~50% w / w, 0.019 mmol, 40%), which was a mixture of approximately 50% w / w with the dimeric hydrolyzed linker. R = 1.46 min (Universal method UPLC); MS (m / z) [M+H] + C 65 H 66 N5O 24 The calculated value of S is 1332.38, and the measured value is 1332.54.

[1122] Example 19

[1123]

[1124] Compound 32 (50 mg, 50% w / w, 0.019 mmol) was dissolved in MeOH:THF 1:1 (1 mL). LiOH (20 mg, 0.84 mmol) was added and the mixture was stirred for 60 minutes. Water (0.5 mL) was added. Complete conversion was observed by UPLC-MS. The reaction was quenched with AcOH, concentrated under vacuum, and purified by preparative HPLC. The fraction containing the desired product was lyophilized to give the desired product, compound 33 (5 mg, 0.005 mmol, 27%), as a yellow solid. LC-MS (Method A): t R =0.84 min; MS(m / z)[M+H] + C 43 H 48 N5O 19 The calculated value of S is 970.27, and the measured value is 969.92.

[1125] Example 20

[1126]

[1127] Compound 33 (5 mg, 0.005 mmol) was dissolved in DMF (0.5 mL). DIPEA (10 μL) was added, followed by N-hydroxysuccinimide 3-(maleimide)-propionic acid (28, 4.1 mg, 0.016 mmol), and the mixture was stirred for 45 min, at which point complete conversion was observed by UPLC-MS. The mixture was quenched with AcOH (20 μL) and purified by preparative HPLC 10 mm Max-RP C12 5-60-95% MeCN H2O solution. The fraction containing the desired product compound 34 was lyophilized to give a yellow powder (2.33 mg, 2.08 μmol, 40.3%). LC-MS (method): t R = 1.35 min; MS (m / z) [M+H] + C 50 H 53 N6O 22 The calculated value of S is 1121.29, and the measured value is 1121.25.

[1128] Example 21

[1129]

[1130] Compound 35 (2.00 g, 4.12 mmol), prepared according to the method in Bioconjugate Chem. (2006) 17:831-840, was dissolved in anhydrous DCM (20 mL). DIPEA (3.59 mL, 20.60 mmol) was added, followed by ...

Claims

1. A method having the formula L-(QD) p Camptothecin conjugates or salts thereof, wherein L represents a ligand unit, wherein the ligand unit is an antibody or its antigen-binding fragment that selectively binds to cancer cell antigens; The subscript p is an integer from 1 to 16; -QD is It is marked with a single asterisk. The wavy line represents the point covalently connected to Z; Z is the extended sub-unit, where Z is , or , Marked with a double asterisk The wavy lines represent sites that are covalently connected to the rest of the QD. Marked with a three-star symbol The wavy lines represent the points covalently bonded to the sulfur atoms of L; and R 17 -C1-C 10 Alkylene-C(=O)-.

2. The camptothecin conjugate or its salt according to claim 1, wherein R 17 -(CH2) 2-5 -C(=O)-.

3. The camptothecin conjugate or its salt according to claim 1, wherein Z is... Optionally having a succinimide ring in hydrolyzed form as the succinamide moiety, wherein the ring is marked with a double asterisk. The wavy lines indicate sites covalently connected to the rest of the QD; and the three-star markings indicate sites... The wavy line represents the point where the sulfur atom of L is covalently bonded.

4. The camptothecin conjugate or its salt according to claim 1, wherein Z is... .

5. The camptothecin conjugate or its salt according to claim 1, wherein Z is... .

6. The camptothecin conjugate or its salt according to claim 1, wherein Z is... .

7. The camptothecin conjugate or its salt according to claim 1, wherein -QD- has the following formula: It is marked with a single asterisk. The wavy line represents the point covalently connected to L.

8. A camptothecin-linker compound or a salt thereof, having the following structure: in Marked with a single asterisk The wavy line represents the point covalently connected to Z'; Z' is the precursor of the extended subunit, where Z' is ， in: Marked with a double asterisk The wavy lines represent sites that are covalently connected to the rest of the camptothecin-linker structure; R 17 -C1-C 10 Alkylene-C(=O)-.

9. The camptothecin-linker compound or a salt thereof according to claim 8, wherein R 17 -(CH2) 2-5 -C(=O)-.

10. The camptothecin-linker compound or its salt according to claim 8, having formula (iii) or formula (x), wherein Z' is , It is marked with a double asterisk. The wavy lines represent sites that are covalently connected to the rest of the camptothecin-linker structure.

11. The camptothecin-linker compound or its salt according to claim 8, having the following structure: , or its salt.

12. Use of camptothecin conjugates in the preparation of medicaments for treating cancer in a subject, wherein the camptothecin conjugate has the formula of claim 1.

13. The use according to claim 12, wherein the cancer is selected from lymphoma, leukemia and solid tumors.

14. The use according to claim 12, wherein the cancer is lymphoma or leukemia.

15. A pharmaceutically acceptable composition comprising the camptothecin conjugate of claim 1 and at least one pharmaceutically acceptable excipient.

16. A composition for treating cancer in a subject who requires it, wherein the composition comprises an effective amount of the camptothecin conjugate of claim 1.

17. The use according to claim 16, wherein the cancer is selected from lymphoma, leukemia and solid tumors.

18. The use according to claim 16, wherein the cancer is lymphoma or leukemia.

19. A method for preparing the camptothecin conjugate of claim 1, the method comprising the step of contacting a targeting agent having a functional group reactive to Z' of the camptothecin-linker compound of claim 8, thereby forming a covalent bond between the ligand unit and the extended subunit (Z) of the camptothecin conjugate, whose structures respectively correspond to the targeting agent and Z', wherein The target is an antibody or its antigen-binding fragment having at least one cysteine ​​residue, wherein the reactive functional group is a thiol and Z' is composed of a maleimide moiety.

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