Bioactive conjugates, methods for preparing the same, and their use

Abstract

This disclosure provides an antibody-drug conjugate platform comprising a conjugator assembly component, and an antibody-drug conjugate comprising a platform-derived linker payload and an antibody or its antigen-binding fragment. In some embodiments, the antibody-drug conjugate is the following formula, or a pharmaceutically acceptable salt, tautomer, solvate, or stereoisomer thereof, where the values ​​of the variables (e.g., BA, U, V, A, a', W, w', Y, y', PA, x) are as described herein. JPEG2026511515000103.jpg53165

Description

[Technical Field]

[0001] 1. Cross-reference of related applications This application claims priority to International Application No. PCT / CN2023 / 083522 (filed March 23, 2023), the disclosures of which are incorporated herein by reference in their entirety.

[0002] 2. Sequence Listing This application includes a sequence listing submitted electronically in XML format, the entire sequence listing of which is incorporated herein by reference. The aforementioned XML copy was created on 14 March 2024, is named 01368-0070-00PCT-ST26, and is 5,851 bytes in size.

[0003] 3. Field This specification provides an antibody-drug conjugate platform, as well as an antibody-drug conjugate (ADC) comprising the platform and an antibody or its antigen-binding fragment, and the use of the ADC platform and ADC. [Background technology]

[0004] 4.Background technology Antibody-drug conjugates (ADCs) combine the targeting effect of an antibody with the cytotoxic activity of a bioactive molecule, creating a "biological missile." ADCs are induced by antibodies to bind to target cells, which then internalize the cells to release a bioactive molecule payload, treating related diseases. The bioactive molecule is covalently bound to the antibody via a linker.

[0005] Lysine is the most common binding site in antibodies, and its ε-amino group can react with the activated carboxyl group of the linker to form an amide bond. Site-directed binding techniques are currently available, in which the carboxyl group of the linker is activated and then forms an amide bond with a specific lysine ε-amino group in the antibody to complete the binding. However, such amide bonds are readily hydrolyzed under enzymatic action in vivo. As a result, the bioactive molecule and antibody dissociate before reaching the target cell, leading to off-target toxicity. The thiol groups of antibody cysteine ​​residues typically exist in the form of disulfide bonds. These disulfide bonds within the antibody can be cleaved to provide multiple free sulfhydryl groups as binding sites. One method of binding to antibody sulfhydryl groups is through a Michael addition reaction between the free sulfhydryl group and an electrophilic maleimide functional group, or through two Michael addition reactions between a specific substrate and the free sulfhydryl group of the antibody to form a unique sulfur crosslink. WO2016142049 discloses amatoxin as a bioactive molecule and a structure containing amatoxin bound to a methylsulfonyl-substituted oxadiazole-based linker, but does not specifically describe the details of antibody binding. As several researchers in the bioconjugate field have pointed out, the thiol substitution product resulting from the reaction between the electrophilic maleimide functional group and the free thiol of the antibody is subject to slow elimination, thus the reaction is reversed. When this reversible reaction occurs in purified ADC preparations, the reaction is barely detectable because the maleimide and thiols regenerated through the elimination process simply react again to reform the intact conjugate. However, in the presence of other thiols, the net effect may be the transfer of maleimide from the antibody of the ADC to other available thiols. This process occurs in plasma, and the transfer of maleimide from ADC to cysteine ​​34 of serum albumin has been documented in the literature (Alley et al., Bioconjugate Chem. 2008, 19, 759-765). This process has also been reported when ADCs are incubated in the presence of excess cysteine ​​or glutathione (Jununtula et al., Nature Biotech, 2012). This disclosure focuses, in particular, on bioconjugates that do not undergo this transfer reaction. [Prior art documents] [Patent Documents]

[0006] [Patent Document 1] International Publication No. 2016142049 [Non-patent literature]

[0007] [Non-Patent Document 1] Alley et al.,Bioconjugate Chem.2008,19,759-765 [Non-Patent Document 2] Jununtula et al.,Nature Biotech,2012 [Overview of the project] [Means for solving the problem]

[0008] 5. Brief Overview This specification provides an antibody-drug conjugate platform and antibody-drug conjugates (ADCs). It also provides the use of the ADC platform for preparing ADCs.

[0009] In some embodiments, the ADC compound of formula (I) is used herein: [ka] Alternatively, a pharmaceutically acceptable salt, tautomer, solvate, or stereoisomer thereof is provided, in the formula, BA is a binder selected from humanized antibodies, chimeric antibodies, human antibodies, or their antigen-binding fragments. U is an arylene, heteroarylene, or bond. V represents a bond or -C≡C-(CH2) n -and, n is an integer between 0 and 10. A is a residue in the stretcher unit, The subscript a' is either 0 or 1. W is a cleaveable unit, The subscript w' is either 0 or 1. Y is a spacer unit, The subscript y' is either 0 or 1. PA is payload residue. The subscript x is between 1 and 15.

[0010] In some embodiments, the platform is a linker-payload compound of formula (II): [ka] or a pharmaceutically acceptable salt, tautomer, solvate, or stereoisomer thereof, in the formula, U is an arylene, heteroarylene, or bond. V represents a bond or -C≡C-(CH2) n -and, n is an integer between 0 and 10. A is a residue in the stretcher unit, The subscript a' is either 0 or 1. W is a cleaveable unit, The subscript w' is either 0 or 1. Y is a spacer unit, The subscript y' is either 0 or 1. PA is a payload residue.

[0011] In some embodiments, the platform is a linker compound of formula (III): [ka] or a pharmaceutically acceptable salt, tautomer, solvate, or stereoisomer thereof, in the formula, U is an arylene, heteroarylene, or bond. V represents a bond or -C≡C-(CH2) n -and, n is an integer between 0 and 10. A is a stretcher unit, The subscript a' is either 0 or 1.

[0012] Further objectives and advantages are partially described in later embodiments for carrying out the invention, and partially understood or known by embodiments for carrying out the invention. These objectives and advantages are realized and achieved by the elements and combinations detailed in the appended claims.

[0013] Please understand that the above general description and the following embodiments for carrying out the invention are merely illustrative and descriptive, and do not limit the scope of the claims.

[0014] The accompanying drawings incorporated herein, and which constitute part thereof, serve to illustrate embodiments and to illustrate the principles described herein, along with modes for carrying out the invention. [Brief explanation of the drawing]

[0015] [Figure 1-1]The stability data for Conjugator-Antibody Conjugate 3-1 is shown in pH 7.4 and 8.0 buffers, with and without GSH. [Figure 1-2] The stability data for Conjugator-Antibody Conjugate 3-1 is shown in pH 7.4 and 8.0 buffers, with and without GSH. [Figure 2-1] The stability data for Conjugator-Antibody Conjugate 3-2 is shown in pH 7.4 and 8.0 buffers, with and without GSH. [Figure 2-2] The stability data for Conjugator-Antibody Conjugate 3-2 is shown in pH 7.4 and 8.0 buffers, with and without GSH. [Figure 3-1] The stability data for Conjugator-Antibody Conjugate 3-3 is shown in pH 7.4 and 8.0 buffers, with and without GSH. [Figure 3-2] The stability data for Conjugator-Antibody Conjugate 3-3 is shown in pH 7.4 and 8.0 buffers, with and without GSH. [Figure 4-1] The stability data for conjugator-antibody conjugates 3-4 are shown in pH 7.4 and 8.0 buffers, with and without GSH. [Figure 4-2] The stability data for conjugator-antibody conjugates 3-4 are shown in pH 7.4 and 8.0 buffers, with and without GSH. [Figure 5-1] The stability data for conjugator-antibody conjugates 3-5 are shown in pH 7.4 and 8.0 buffers, with and without GSH. [Figure 5-2] The stability data for conjugator-antibody conjugates 3-5 are shown in pH 7.4 and 8.0 buffers, with and without GSH. [Figure 6] This shows the stability data of conjugator-antibody conjugate 3-1 after 168 hours in pH 5.5 histidine buffer. [Figure 7]This shows the stability data of conjugator-antibody conjugate 3-2 after 168 hours in pH 5.5 histidine buffer. [Figure 8-1] This shows the stability data of the conjugator-antibody conjugate 3-3 after 168 hours in pH 5.5 histidine buffer. [Figure 8-2] This shows the stability data of the conjugator-antibody conjugate 3-3 after 168 hours in pH 5.5 histidine buffer. [Figure 9-1] The stability data for conjugator-antibody conjugates 3-4 after 168 hours in pH 5.5 histidine buffer is shown. [Figure 9-2] The stability data for conjugator-antibody conjugates 3-4 after 168 hours in pH 5.5 histidine buffer is shown. [Figure 10-1] This shows the stability data of conjugator-antibody conjugates 3-5 after 168 hours in pH 5.5 histidine buffer. [Figure 10-2] This shows the stability data of conjugator-antibody conjugates 3-5 after 168 hours in pH 5.5 histidine buffer. [Figure 11-1] The stability data for ADC 4-1 in GSH buffer at pH 7.4 or 8.0 is shown. [Figure 11-2] The stability data for ADC 4-1 in GSH buffer at pH 7.4 or 8.0 is shown. [Figure 12-1] The stability data for ADC 4-2 in GSH buffer at pH 7.4 or 8.0 is shown. [Figure 12-2] The stability data for ADC 4-2 in GSH buffer at pH 7.4 or 8.0 is shown. [Figure 13-1] The stability data for ADC 4-3 in GSH buffer at pH 7.4 or 8.0 is shown. [Figure 13-2] The stability data for ADC 4-3 in GSH buffer at pH 7.4 or 8.0 is shown. [Figure 14-1] This shows the stability data of ADC 4-1 in formulation buffer. [Figure 14-2] This shows the stability data of ADC 4-1 in formulation buffer. [Figure 15-1] This shows the stability data of ADC 4-2 in formulation buffer. [Figure 15-2] This shows the stability data of ADC 4-2 in formulation buffer. [Figure 16-1] This shows the stability data of ADC 4-3 in formulation buffer. [Figure 16-2] This shows the stability data of ADC 4-3 in formulation buffer. [Figure 17] This demonstrates the direct killing activity of ADC against HL60 cells. [Figure 18] This demonstrates the direct killing activity of ADC against U937 cells. [Figure 19] This demonstrates the direct killing activity of ADC against TF1 cells. [Figure 20] ADC has shown direct killing activity against B7H3-highly expressing cell lines (H1650). [Figure 21] ADC has shown direct killing activity against B7H3-low-expressing cell lines (Capan-1). [Figure 22] This study demonstrates the direct killing activity of ADC against the B7H3(-) cell line (MB-453). [Figure 23] ADC has shown direct killing activity against B7H3-highly expressing cell lines (H1650). [Figure 24] ADC has shown direct killing activity against B7H3-low-expressing cell lines (Capan-1). [Figure 25] This study demonstrates the direct killing activity of ADC against the B7H3(-) cell line (MB-453). [Figure 26] This shows bystander killing of ADCs against NCI-H358 / MDA-MB-453 (nano-Luc) in a co-culture assay. [Figure 27]This shows bystander killing of ADCs against NCI-H358 / MDA-MB-453 (nano-Luc) in a co-culture assay. [Figure 28] This shows bystander killing of ADCs against NCI-H358 / MDA-MB-453 (nano-Luc) in a co-culture assay. [Figure 29] This shows bystander killing of ADCs against NCI-H358 / MDA-MB-453 (nano-Luc) in a co-culture assay. [Figure 30] This line graph shows the antitumor activity of ADCs in an H1650 xenograft model. [Figure 31] This line graph shows the dose-dependent antitumor activity of ADCs in an H1650 xenograft model. [Figure 32] This is a line graph showing the payload release rate from ADCs in mouse plasma. [Figure 33] This is a line graph showing the payload release rate from ADCs in human plasma. [Figure 34] This is a line graph showing the payload release rate from ADCs in mouse plasma. [Figure 35] This is a line graph showing the payload release rate from ADCs in human plasma. [Figure 36] This is a line graph showing the PK profile of ADC in tumor-bearing mice. [Figure 37] This is a line graph showing the PK profile of ADC in non-tumor-bearing mice. [Modes for carrying out the invention]

[0016] 7. Detailed explanation This specification provides antibody-drug conjugates (ADCs), as well as covalent linkers and linker payloads (platforms) for producing ADCs. ADCs can be used to treat diseases or disorders, such as cancer, by providing compositions containing ADCs. The ADCs disclosed herein are more stable than known ADCs.

[0017] Some conjugators are known to be difficult to conjugate with binders, for example, requiring harsh reaction conditions that can adversely modify the binder or requiring multiple reaction steps. The conjugator assembly of this disclosure efficiently conjugates to cysteine ​​residues on antibodies without requiring, for example, separate hydrolysis steps. Some ADCs are known to undergo deconjugation under physiological conditions. The conjugator-antibody conjugate of this disclosure is less prone to early deconjugation in plasma because the binding of the conjugator assembly to the binder and payload is improved.

[0018] 7.1.Definition In this disclosure, the following terms have the meanings set forth below unless otherwise indicated. Unless otherwise defined, all technical and scientific terms used herein have the meanings commonly understood by those skilled in the art in which this disclosure is made. If there are multiple definitions of a term used herein, the definitions in this section shall prevail unless otherwise noted.

[0019] In this specification, the term “antibody” is used in its broadest sense and specifically encompasses intact monoclonal antibodies, polyclonal antibodies, monospecific antibodies, multispecific antibodies (e.g., bispecific antibodies), and antibody fragments exhibiting desired biological activity. Intact antibodies primarily have two regions: a variable region and a constant region. The variable region binds to and interacts with the target antigen. The variable region includes a complementary determination region (CDR) that recognizes and binds to a specific binding site on a particular antigen. The constant region is recognized by the immune system and can interact with it (see, e.g., Janeway et al., 2001, Immuno. 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, the antibody is derived from a human or mouse. The antibody may be, for example, a human, humanized, or chimeric antibody.

[0020] As used herein, the term “monoclonal antibody” refers to an antibody obtained from a substantially homogeneous antibody population (i.e., the individual antibodies constituting the population are identical except for minor naturally occurring variations that may be present). Monoclonal antibodies are highly specific and target a single antigenic site. The modifier “monoclonal” should not be interpreted as requiring antibody production by any particular method.

[0021] An "intact antibody" is an antibody that, in addition to an antigen-binding variable region, contains a light chain constant domain (CL) and a heavy chain constant domain (CH1, CH2, CH3, and CH4) (depending on the antibody class). The constant domain may be a native sequence constant domain (e.g., a human native sequence constant domain) or an amino acid sequence variant thereof.

[0022] "Antibody fragment" includes a part of an intact antibody that contains an antigen-binding region or variable region. Examples of antibody fragments include Fab, Fab’, F(ab’)2, and Fv fragments, diabodies, triabodies, tetrabodies, linear antibodies, single-chain antibody molecules, scFv, scFv-Fc, antibody fragment(s), multispecific antibody fragments formed from fragment(s) produced by a Fab expression library, or any of the above epitope-binding fragments that immunospecifically bind to a target antigen (e.g., a cancer cell antigen, a viral antigen, or a microbial antigen).

[0023] "Antigen" is an entity to which an antibody specifically binds.

[0024] The terms "specific binding" and "specifically binds" mean that an antibody or antibody derivative binds in a highly selective manner to its corresponding target antigen and does not bind to a large number of other antigens. Typically, an antibody or antibody derivative binds with an affinity of at least about 1×10 -7 M, 10 -8 M, 10 -9 M, 10 -10 M, 10 -11 M, or 10 -12 M and binds to a given antigen with an affinity that is at least 2-fold greater than the affinity in binding to a non-specific antigen (e.g., BSA, casein) other than the given antigen or an antigen closely related thereto.

[0025] The terms "inhibit" or "inhibition of" mean to reduce a measurable amount or to prevent completely.

[0026] The term "therapeutic dose" refers to the amount of a drug effective in treating a disease or disorder in a mammal. In the case of cancer, a therapeutic dose of a drug can reduce the number of cancer cells, shrink tumor size, inhibit (i.e., slow or stop to some extent) cancer cell invasion into peripheral organs, inhibit (i.e., slow or stop to some extent) tumor metastasis, inhibit tumor growth to some extent, and / or alleviate to some extent one or more symptoms associated with cancer. A drug has cell division inhibitory and / or cytotoxic properties to the extent that it can inhibit growth and / or kill existing cancer cells. For cancer treatment, efficacy can be measured, for example, by evaluating the time to progression (TTP) and / or quantifying the response rate (RR).

[0027] The terms “considerable” or “considerable” refer to a majority of the mixture or sample, i.e., more than 50% of the population (for example, more than 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% of the population).

[0028] The terms "intracellularly cleaved" and "intracellular cleavage" refer to an intracellular metabolic process or reaction of a ligand-drug conjugate (e.g., antibody-drug conjugate (ADC)) in which the covalent bond (e.g., linker) between the drug portion (D) and the ligand unit (e.g., antibody (BA or Ab)) is cleaved, resulting in the release of the drug from the antibody or the dissociation of another metabolite of the conjugate within the cell. Therefore, the cleaved portion of a drug-linker-ligand conjugate is an intracellular metabolite.

[0029] The term "cytotoxic activity" refers to the cytotoxic, cell division inhibiting, or antiproliferative activity of a drug-linker-ligand conjugate compound or its intracellular metabolite. Cytotoxic activity can be expressed as an IC50 value, which is the concentration (moles or mass) per unit volume at which half of the cells survive.

[0030] As used herein, the term “cytotoxic agent” refers to a substance that inhibits cellular function and / or causes cellular destruction. This term is intended to include radioisotopes (e.g., radioisotopes of 211At, 131I, 125I, 90Y, 186Re, 188Re, 153Sm, 212Bi, 32P, 60C, and Lu), chemotherapeutic agents, and toxins (e.g., low-molecular-weight toxins or enzymatic toxins of bacterial, fungal, plant, or animal origin) (including their synthetic analogs and derivatives).

[0031] The terms “cancer” and “malignant” refer to or represent a physiological condition or disorder in mammals typically characterized by uncontrolled cell growth. A “tumor” contains one or more cancerous cells. Examples of cancer include, but are not limited to, carcinomas, lymphomas, blastomas, sarcomas, and leukemias or lymphoid malignancies. More specific examples of such cancers include squamous cell carcinoma (e.g., epithelial squamous cell carcinoma), lung cancer (including small cell lung cancer, non-small cell lung cancer ("NSCLC"), lung adenocarcinoma, and lung squamous cell carcinoma), peritoneal cancer, hepatocellular carcinoma, gastric cancer or stomach cancer including gastrointestinal cancer, pancreatic cancer, glioblastoma, cervical cancer, ovarian cancer, liver cancer, bladder cancer, hepatocellular carcinoma, breast cancer, colon cancer, rectal cancer, colorectal cancer, endometrial cancer or uterine cancer, salivary gland cancer, kidney cancer or renal cancer, prostate cancer, vulvar cancer, thyroid cancer, liver cancer, anal cancer, penile cancer, and head and neck cancer.

[0032] In this specification, "autoimmune disease" refers to a disease or disorder that originates from and is directed against the tissues or proteins of an individual.

[0033] Examples of “patients” include, but are not limited to, mammals (e.g., humans, rats, mice, guinea pigs, monkeys, pigs, goats, cattle, horses, dogs, or cats), and birds or poultry. In one embodiment, the patient is a human.

[0034] The terms “to treat” or “treatment” refer to therapeutic treatment and preventive measures to prevent recurrence, unless otherwise indicated by the context, and their purpose is to suppress or slow (mitigate) undesirable physiological changes or impairments (e.g., the development or spread of cancer). For the purposes of this disclosure, beneficial or desirable clinical outcomes include, but are not limited to, symptom relief, reduction of disease severity, stabilization of the disease state (i.e., no worsening), delay or slowing of disease progression, improvement or mitigation of the disease state, and remission (partial or overall), whether detectable or undetectable. “Treatment” may also mean extending survival compared to the survival expected without treatment. Those requiring treatment include those who already have a condition or impairment, as well as those who are prone to developing a condition or impairment.

[0035] In the context of cancer, the term “to treat” includes any or all of the following: inhibiting tumor cells, cancer cells, or tumor growth; inhibiting the replication of tumor cells or cancer cells; reducing the overall tumor burden or decreasing the number of cancer cells; and improving one or more symptoms associated with the disease.

[0036] In the context of autoimmune diseases, the term “treat” includes any or all of the following: inhibiting the replication of cells associated with the autoimmune disease (including, but not limited to, cells that produce autoimmune disease antibodies); reducing the autoimmune antibody load; and improving one or more symptoms of the autoimmune disease.

[0037] As used herein and in the appended claims, the indefinite articles "a" and "an," and the definite article "the," include singular as well as plural referents, unless otherwise explicitly indicated by the context.

[0038] As used herein, and unless otherwise specified, the terms “about” and “approximately” mean, when used in relation to the amount or weight percentage of a component of a composition, an amount or weight percentage that would be recognized by those skilled in the art as producing an equivalent pharmacological effect to that obtained from a specified amount or weight percentage. In certain embodiments, when used in this context, the terms “about” and “approximately” mean an amount or weight percentage that is within 30%, 20%, 15%, 10%, or 5% of a specified amount or weight percentage.

[0039] As used herein, unless otherwise specified, the terms “about” and “approximately” mean, when used in relation to a numerical value or range of values ​​provided to characterize a particular solid form, e.g., a specific temperature or temperature range, e.g., one describing melting, dehydration, desolvation, or glass transition temperature; a mass change, e.g., a mass change as a function of temperature or humidity; a solvent or water content (e.g., in terms of mass or percentage); or a peak position, e.g., in analysis by IR, Raman spectroscopy, or XRPD, that the value or range of values ​​may deviate to an extent that is reasonable to those skilled in the art, but still describe a solid form. Techniques for characterizing crystalline and amorphous solids include, but are not limited to, thermogravimetric analysis (TGA), differential scanning calorimetry (DSC), X-ray powder diffraction (XRPD), single-crystal X-ray diffraction, vibrational spectroscopy, e.g., infrared (IR) and Raman spectroscopy, solid-state and solution nuclear magnetic resonance (NMR) spectroscopy, optical microscopy, hot-stage optical microscopy, scanning electron microscopy (SEM), electron crystallography and quantitative analysis, particle size analysis (PSA), surface area analysis, solubility testing, and dissolution testing. In certain embodiments, the terms “about” and “approximately” used in this context indicate that a numerical value or range may vary within 30%, 20%, 15%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1.5%, 1%, 0.5%, or 0.25% of the given numerical value or range. For example, in some embodiments, the value of the XRPD peak position may vary by up to ±0.2°²θ while describing a particular XRPD peak.

[0040] The "alkyl" group is a saturated, partially saturated, or unsaturated linear or branched acyclic hydrocarbon having 1 to 10 carbon atoms, typically 1 to 8 carbon atoms, and in some embodiments 1 to 6, 1 to 4, or 2 to 6 carbon atoms. Representative alkyl groups include -methyl, -ethyl, -n-propyl, -n-butyl, -n-pentyl, and -n-hexyl. Saturated branched alkyl groups include -isopropyl, -sec-butyl, -isobutyl, -tert-butyl, -isopentyl, 2-methylpentyl, 3-methylpentyl, 4-methylpentyl, and 2,3-dimethylbutyl. Examples of unsaturated alkyl groups include, but are not limited to, vinyl, allyl, -CH=CH(CH3), -CH=C(CH3)2, -C(CH3)=CH2, -C(CH3)=CH(CH3), C(CH2CH3)=CH2, -C≡CH, -C≡C(CH3), -C≡C(CH2CH3), -CH2C≡CH, -CH2C≡C(CH3), and CH2C≡C(CH2CH3). Alkyl groups may be substituted or unsubstituted. In certain embodiments, where an alkyl group described herein is expressed as "substituted," the alkyl group may be substituted with halogens (chloro, iodine, bromo, or fluoro), hydroxyl, alkoxy, alkoxyalkyl, amino, alkylamino, carboxy, nitro, cyano, thiol, thioether, imine, imide, amidine, guanidine, enamine, aminocarbonyl, acylamino, phosphonato, phosphine, thiocarbonyl, sulfonyl, sulfone, sulfonamide, ketone, aldehyde, ester, urea, urethane, oxime, hydroxylamine, alkoxyamine, aralkoxyamine, N-oxide, hydrazine, hydrazide, hydrazone, azide, isocyanate, isothiocyanate, cyanate, thiocyanate, B(OH)2, or O(alkyl)aminocarbonyl, in addition to any substituent(s) shown in the compounds and embodiments disclosed herein.

[0041] The "alkenyl" group is a linear or branched acyclic hydrocarbon having 2 to 10 carbon atoms, typically 2 to 8 carbon atoms, and containing at least one carbon-carbon double bond. Representative linear and branched (C2-C8) alkenyls include -vinyl, -allyl, -1-butenyl, -2-butenyl, -isobutyrenyl, -1-pentenyl, -2-pentenyl, -3-methyl-1-butenyl, -2-methyl-2-butenyl, -2,3-dimethyl-2-butenyl, -1-hexenyl, 2-hexenyl, -3-hexenyl, -1-heptenyl, -2-heptenyl, -3-heptenyl, -1-octenyl, -2-octenyl, and 3-octenyl. The double bond of the alkenyl group may or may not conjugate with another unsaturated group. The alkenyl group may be unsubstituted or substituted.

[0042] A "cycloalkyl" group is a saturated or partially saturated cyclic alkyl group of 3 to 10 carbon atoms having a single cyclic ring or multiple fused or crosslinked rings, which may optionally be substituted with 1 to 3 alkyl groups. In some embodiments, the cycloalkyl group has 3 to 8 ring members, while in other embodiments, the number of ring carbon atoms ranges from 3 to 5, 3 to 6, or 3 to 7. Examples of such cycloalkyl groups include monocyclic structures (e.g., cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, cyclooctyl, 1-methylcyclopropyl, 2-methylcyclopentyl, 2-methylcyclooctyl, etc.) or polycyclic or crosslinked ring structures (e.g., adamantyl, etc.). Examples of unsaturated cycloalkyl groups include, in particular, cyclohexenyl, cyclopentenyl, cyclohexadienyl, butadienyl, pentadienyl, and hexadienyl. Cycloalkyl groups may be substituted or unsubstituted. Examples of such substituted cycloalkyl groups include cyclohexanone.

[0043] An "aryl" group is an aromatic carbocyclic group with 6 to 14 carbon atoms, having a single ring (e.g., phenyl) or multiple fused rings (e.g., naphthyl or anthryl). In some embodiments, the aryl group contains 6 to 14 carbon atoms in the ring portion of the group, and in others 6 to 12, and possibly 6 to 10 carbon atoms. Specific examples of aryls include phenyl, biphenyl, and naphthyl. Aryl groups can be substituted or unsubstituted. The term "aryl group" also includes groups containing fused rings, such as fused aromatic aliphatic ring systems (e.g., indanyl, tetrahydronaphthyl, etc.).

[0044] The "arylene" group is a divalent aryl group as defined herein.

[0045] A "heteroaryl" group is an aryl ring system having 1 to 4 heteroatoms as ring atoms in a heteroaromatic ring system, with the remaining atoms being carbon atoms. In some embodiments, the heteroaryl group contains 5 to 6 ring atoms in the ring portion of the group, and in other cases, 6 to 9 or 6 to 10 atoms. Suitable heteroatoms include oxygen, sulfur, and nitrogen. In certain embodiments, the heteroaryl ring system is monocyclic or bicyclic. Non-limiting examples include, but are not limited to, pyrrolyl, pyrazolyl, imidazolyl, triazolyl, tetrazolyl, oxazolyl, isoxazolyl, thiazolyl, pyrrolyl, pyridyl, pyridadinyl, pyrimidinyl, pyrazinyl, thiophenyl, benzothiophenyl, furanyl, benzofuranyl (e.g., isobenzofuran-1,3-diimine), indolyl, azaindolyl (e.g., pyrrolopyridyl or 1H-pyrrolo[2,3-b]pyridyl), indazolyl, benzimidazolyl (e.g., 1H-benzo Examples of groups include [d]imidazolyl), imidazopyridyl (e.g., azabenzimidazolyl, 3H-imidazo[4,5-b]pyridyl, or 1H-imidazo[4,5-b]pyridyl), pyrazolopyridyl, triazolopyridyl, benzotriazolyl, benzoxazolyl, benzothiazolyl, benzothiadiazolyl, isoxazolopyridyl, thianaphthalenyl, prinyl, xanthinyl, adeninyl, guaninyl, quinolinyl, isoquinolinyl, tetrahydroquinolinyl, quinoxalinyl, and quinazolinyl groups.

[0046] A "heteroarylene" group is a divalent heteroaryl group as defined herein.

[0047] A "heterocyclyl" is an aromatic (also called heteroaryl) or non-aromatic cycloalkyl group in which 1 to 4 of the ring carbon atoms are independently replaced by heteroatoms from the group consisting of O, S, and N. In some embodiments, the heterocyclyl group contains 3 to 10 ring members, while other such groups have 3 to 5, 3 to 6, or 3 to 8 ring members. Furthermore, the heterocyclyl may be bonded to other groups by any ring atom (i.e., any carbon or heteroatom of the heterocyclic ring). The heteroaryl group may be substituted or unsubstituted. Heterocyclyl groups include unsaturated, partially saturated, and saturated ring systems (e.g., imidazolyl, imidazolinyl, and imidazolidinyl groups). The term "heterocyclyl" includes fused ring species (including ring species containing fused aromatic and non-aromatic groups), such as benzotriazolyl, 2,3-dihydrobenzo[1,4]dioxynyl, and benzo[1,3]dioxolyl. This term also includes bridging polycyclic ring systems containing heteroatoms (for example, quinuclidyls, but not limited to them).Examples of heterocyclyl groups, though not limited to them, include azilidinyl, azetidinyl, pyrrolidyl, imidazolidinyl, pyrazolidinyl, thiazolidinyl, tetrahydrothiophenyl, tetrahydrofuranil, dioxolyl, furanil, thiophenyl, pyrrolyl, pyrrolinil, imidazolyl, pyrazolyl, pyrazolinil, triazolyl, tetrazolyl, oxazolyl, isoxazolyl, thiazolyl, thiazolinyl, isothiazolyl, thiadiazolyl, oxadiazolyl, piperidyl, piperazinyl, morpholinil, and thiomorpholinyl. Tetrahydropyranil (e.g., tetrahydro-2H-pyranil), tetrahydrothiopyranil, oxatian, dioxyl, dithianil, pyranil, pyridyl, pyrimidinil, pyridazinil, pyrazinil, triazinil, dihydropyridyl, dihydrodithinyl, dihydrodithionyl, homopiperazinil, quinuclidyl, indolyl, indolinyl, isoindolyl, azaindolyl (pyrrolopyridyl), indazolyl, indolidinyl, benzotriazolyl, benzimidazolyl, benzofuranil, benzothiophenyl, benzothiazolyl Benzoxadiazolyl, benzoxazinyl, benzodithinyl, benzoxathinyl, benzothiadinyl, benzoxazolyl, benzothiazolyl, benzothiadiazolyl, benzo[1,3]dioxolyl, pyrazolopyridyl, imidazopyridyl (azabenzimidazolyl, e.g., 1H-imidazo[4,5-b]pyridyl or 1H-imidazo[4,5-b]pyridin-2(3H)-onyl), triazolopyridyl, isoxazolopyridyl, prinyl, xanthinyl, adeninyl, guaninyl, quinolinyl, isoquinolinyl, ki Examples include noridinyl, quinoxalinyl, quinazolinyl, cinolinyl, phthalazinyl, naphthilidinyl, pteridinyl, thianaphthalenyl, dihydrobenzothiadinyl, dihydrobenzofuranyl, dihydroindolyl, dihydrobenzodioxynyl, tetrahydroindolyl, tetrahydroindazolyl, tetrahydrobenzimidazolyl, tetrahydrobenzotriazolyl, tetrahydropyrrolopyridyl, tetrahydropyrazolopyridyl, tetrahydroimidazopyridyl, tetrahydrotriazolopyridyl, and tetrahydroquinolinyl groups.Typical substituted heterocyclyl groups may be monosubstituted or multiple substituted, such as pyridyl or morpholinyl groups that are 2-, 3-, 4-, 5-, or 6-substituted, or disubstituted with various substituents as listed below.

[0048] The "cycloalkylalkyl" group is a radical of the formula:-alkyl-cycloalkyl (wherein alkyl and cycloalkyl are defined above). A substituted cycloalkylalkyl group may be substituted with the alkyl moiety, the cycloalkyl moiety, or both the alkyl and cycloalkyl moieties of the group. Representative cycloalkylalkyl groups, but not limited to, include cyclopentylmethyl, cyclopentylethyl, cyclohexylmethyl, cyclohexylethyl, and cyclohexylpropyl. Representative substituted cycloalkylalkyl groups may be monosubstituted or multiple substituted.

[0049] The "aralkyl" group is a radical of the formula :-alkyl-aryl (wherein alkyl and aryl are defined above). A substituted aralkyl group may be substituted with the alkyl moiety, the aryl moiety, or both the alkyl and aryl moieties. Representative aralkyl groups, but not limited to, include benzyl and phenethyl groups, as well as condensed (cycloalkylaryl)alkyl groups (e.g., 4-ethyl-indanyl).

[0050] The "heterocyclylalkyl" group is a radical of the formula :-alkyl-heterocyclyl (wherein alkyl and aryl are defined above). A substituted heterocyclylalkyl group may be substituted with the alkyl moiety, the heterocyclyl moiety, or both the alkyl and heterocyclyl moieties of the group. Representative heterocyclylalkyl groups include, but are not limited to, 4-ethyl-morpholinyl, 4-propylmorpholinyl, furan-2-ylmethyl, furan-3-ylmethyl, pyrzin-3-ylmethyl, (tetrahydro-2H-pyran-4-yl)methyl, (tetrahydro-2H-pyran-4-yl)ethyl, tetrahydrofuran-2-ylmethyl, tetrahydrofuran-2-ylethyl, and indole-2-ylpropyl.

[0051] "Halogen" refers to chloro, iodine, bromo, or fluoro compounds.

[0052] A "hydroxyalkyl" group is one in which the aforementioned alkyl group is replaced with one or more hydroxyl groups.

[0053] The "alkoxy" group is O(alkyl) (where alkyl is defined as vapor).

[0054] The "alkoxyalkyl" group is -(alkyl)-O-(alkyl) (where alkyl is defined above).

[0055] As used herein, “alkynyl” refers to a monovalent hydrocarbon radical moiety comprising at least two carbon atoms and one or more carbon-carbon triple bonds. Alkynnyls can be optionally substituted and may be linear, branched, or cyclic. Alkynnyls are, but are not limited to, radicals having 2 to 20 carbon atoms, i.e., C 2~20 Alkynnyl, a radical having 2 to 12 carbon atoms, i.e., C 2~12 Alkynnyl, a radical having 2 to 8 carbon atoms, i.e., C 2~8 Alkynnyl, a radical having 2 to 6 carbon atoms, i.e., C 2~6Alkynnyls and radicals having 2 to 4 carbon atoms, i.e., C 2~4 Alkynnyl is one example. Examples of alkynyl moieties, though not limited to them, include ethynyl, propynyl, and butynyl.

[0056] As used herein, “haloalkyl” means the alkyl group as defined above, where the alkyl group comprises at least one substituent selected from halogens, such as fluorine (F), chlorine (Cl), bromine (Br), or iodine (I). Examples of haloalkyl groups, but not limited to, include -CF3, -CH2CF3, -CCl2F, and -CCl3.

[0057] As used herein, “haloalkoxy” means the alkoxy as defined above, where the alkoxy comprises at least one substituent selected from a halogen, e.g., F, Cl, Br, or I.

[0058] As used herein, “arylalkyl” refers to the monovalent radical portion of an alkyl compound, and the alkyl compound is substituted with an aromatic substituent. That is, the aromatic compound contains a single bond to the alkyl group, and the radical is localized to the alkyl group. The arylalkyl group is bonded to the shown chemical structure via the alkyl group. The arylalkyl group can be represented by the structure, for example, B-CH2-, B-CH2-CH2-, B-CH2-CH2-CH2-, B-CH2-CH2-CH2-CH2-, B-CH(CH3)-CH2-CH2-, B-CH2-CH(CH3)-CH2-, where B is an aromatic moiety, for example, phenyl. The arylalkyl group is optionally substituted; that is, the aryl group and / or alkyl group can be substituted as disclosed herein. Examples of arylalkyl groups, but not limited to, benzyl.

[0059] As used herein, “alkylaryl” refers to the monovalent radical portion of an aryl compound, where the aryl compound is substituted with an alkyl substituent. That is, the aryl compound contains a single bond to the alkyl group, and the radical is localized to the aryl group. The alkylaryl group is bonded to the shown chemical structure via the aryl group. Alkylaryls can be represented by structures such as -B-CH3, -B-CH2-CH3, -B-CH2-CH2-CH3, -B-CH2-CH2-CH2-CH3, -B-CH(CH3)-CH2-CH3, and -B-CH2-CH(CH3)-CH3, where B is an aromatic moiety, such as phenyl. Alkylaryls are optionally substituted; that is, the aryl group and / or alkyl group can be substituted as disclosed herein. Examples of alkylaryls include, but are not limited to, toluyl.

[0060] As used herein, "aryloxy" refers to the monovalent radical portion of an aromatic compound, where the ring atom is a carbon atom and the ring is substituted with an oxygen radical; that is, the aromatic compound contains a single bond to the oxygen atom and the radical is localized to the oxygen atom (e.g., in the case of phenoxy, C6H5-O-). The aryloxy substituent bonds to the compound it substitutes through this oxygen atom. The aryloxy is optionally substituted. The aryloxy can be any radical having 6 to 20 ring carbon atoms, i.e., C6H5-O-. 6~20 Aryloxy radicals are radicals having 6 to 15 ring carbon atoms, i.e., C 6~15 Aryloxy radicals, and radicals having 6-10 ring carbon atoms, i.e., C 6~10 Examples include aryloxy radicals. Examples of aryloxy parts include, but are not limited to, phenoxy, naphthoxy, and anthroxy.

[0061] The "amino" group is a radical with the formula:NH2.

[0062] The "hydroxylamine" group is represented by formula: N(R #) is a radical of OH or NHOH (where R is the radical in the formula). # (wherein this specification is a substituted or unsubstituted alkyl, cycloalkyl, cycloalkylalkyl, aryl, aralkyl, heterocyclyl, or heterocyclylalkyl group).

[0063] The "alkoxyamine" group is represented by the formula: -N(R # ) an O-alkyl or -NHO-alkyl radical (wherein R is the radical in the formula) # (This is as defined above).

[0064] The "aralcoxyamine" group is represented by formula: N(R # ) an O-aryl or NHOaryl radical (wherein R is the radical in the formula) # (This is as defined above).

[0065] The "alkylamine" group is a radical of the formula:NH alkyl or N(alkyl)2 (wherein each alkyl is independently as defined above).

[0066] The "aminocarbonyl" group is represented by the formula: -C(=O)N(R # )2, -C(=O)NH(R # ), or a radical of C(=O)NH2 (wherein each R # (This is as defined above).

[0067] The "acylamino" group is represented by the formula: NHC(=O)(R # ) or N(alkyl)C(=O)(R # ) is a radical (wherein each alkyl and R # (These are independent, as defined above.)

[0068] The "O(alkyl)aminocarbonyl" group has the formula: -O(alkyl)C(=O)N(R # )2,-O(alkyl)C(=O)NH(R # ), or a radical of -O(alkyl)C(=O)NH2 (wherein each R, #(These are independent, as defined above.)

[0069] The "N-oxide" group is represented by the formula: -N + -O - It is radical.

[0070] The "carboxyl" group is a radical with the formula: -C(O)OH.

[0071] The "ketone" group is represented by the formula: C(=O)(R # ) is a radical (where R is in the formula) # (This is as defined above).

[0072] The "aldehyde" group is a radical with the formula -CH(=O).

[0073] The "ester" group is represented by the formula: C(=O)O(R # ) or OC (=O) (R # ) is a radical (where R is in the formula) # (This is as defined above).

[0074] The "urea" group has the formula: -N(alkyl)C(=O)N(R # )2,-N(alkyl)C(=O)NH(R # ), -N(alkyl)C(=O)NH2, -NHC(=O)N(R # )2, -NHC(=O)NH(R # ), or NHC(=O)NH2 # It is a radical of (wherein each alkyl and R # (These are independent, as defined above.)

[0075] The "imine" group is given by the formula: -N=C(R # )2 or -C(R # )=N(R # ) is a radical (where each R # (These are independent, as defined above.)

[0076] The "imide" group is represented by the formula: -C(=O)N(R#)C(=O)(R# ) or N((C=O)(R # ))2 radicals (where each R # (These are independent, as defined above.)

[0077] The "urethane" group is represented by the formula: -OC(=O)N(R # )2, -OC(=O)NH(R # ), -N(R # )C(=O)O(R # ), or -NHC(=O)O(R # ) is a radical (where each R # (These are independent, as defined above.)

[0078] The "amidine" group is represented by the formula: -C(=N(R # ))N(R # )2, -C(=N(R # ))NH(R # ), -C(=N(R # ))NH2, -C(=NH)N(R # )2, -C(=NH)NH(R # ), -C(=NH)NH2, -N=C(R # )N(R # )2, -N=C(R # )NH(R # ), -N=C(R # )NH2, -N(R # )C(R # )=N(R # ), -NHC(R # )=N(R # ), -N(R # )C(R # )=NH, or -NHC(R # )=NH radical (where R # (These are independent, as defined above.)

[0079] The "guanidine" group is represented by the formula: -N(R # )C(=N(R # ))N(R # )2, -NHC(=N(R # ))N(R # )2, -N(R# )(C(=NH)N(R # ))2, -N(R # )(C(=N(R # ))NH(R # ), -N(R # )(C(=N(R # ))NH2, -NHC(=NH)N(R # ))2, -NHC(=N(R # ))NH(R # ), -NHC(=N(R # ))NH2, -NHC(=NH)NH(R # ), -NHC(=NH)NH2, -N=C(N(R # ))2)2, -N=C(NH(R # ))2, or -N=C(NH2)2 radical (where R # is independently as defined above).

[0080] The "enamine" group has the formula: -N(R # )(C(R # )=C(R # ))2, -NHC(R # )=C(R # ))2, -C(N(R # ))2)=C(R # ))2, -C(NH(R # ))=C(R # ))2, -C(NH2)=C(R # ))2, -C(R # )=C(R # )(N(R # ))2), C(R # )=C(R # )(NH(R # ))), or -C(R # )=C(R # )(NH2) radical (where R # is independently as defined above).

[0081] The "oxime" group has the formula: -C(=NO(R # ))(R # ), -C(=NOH)(R # ), -CH(=NO(R #)), or a radical of -CH(=NOH) (where each R # (These are independent, as defined above.)

[0082] The "hydrazide" group is represented by the formula: -C(=O)N(R # )N(R # )2, -C(=O)NHN(R # )2, -C(=O)N(R # )NH(R # ), -C(=O)N(R # )NH2, -C(=O)NHNH(R # )2, or the radical of -C(=O)NHNH2 (where R is the radical in the formula) # (These are independent, as defined above.)

[0083] The "hydrazine" group is represented by the formula: -N(R # )N(R # )2, -NHN(R # )2, -N(R # )NH(R # ) 、 -N(R # )NH2, -NHNH(R # )2, or the radical of -NHNH2 (wherein R, R # (These are independent, as defined above.)

[0084] The "hydrazone" group is represented by the formula: -C(=NN(R # )2)(R # )2, -C(=NNH(R # ))(R # )2, -C(=N-NH2)(R # )2, -N(R # )(N=C(R # )2), or -NH(N=C(R # )2) is a radical (where R is # (These are independent, as defined above.)

[0085] The "azide" group is a radical of formula -N3.

[0086] The "isocyanate" group is a radical with the formula N=C=O.

[0087] The "isothiocyanate" group is a radical with the formula N=C=S.

[0088] The "cyanate" group is the radical of formula OCN.

[0089] The "thiocyanate" group is a radical of formula SCN.

[0090] The "thioether" group is represented by the formula: -S(R # ) is a radical (where R is in the formula) # (These are independent, as defined above.)

[0091] The "thiocarbonyl" group is represented by the formula: -C(=S)(R # ) is a radical (where R is in the formula) # (These are independent, as defined above.)

[0092] The "sulfinyl" group is represented by the formula: -S(=O)(R # ) is a radical (where R is in the formula) # (These are independent, as defined above.)

[0093] The "sulfone" group is represented by the formula: -S(=O)2(R # ) is a radical (where R is in the formula) # (These are independent, as defined above.)

[0094] The "sulfonylamino" group is represented by the formula: -NHSO2(R # ) or -N(alkyl)SO2(R # ) is a radical (wherein each alkyl and R # (as defined above).

[0095] The "sulfonamide" group is represented by the formula: -S(=O)2N(R # )2, -S(=O)2NH(R # ), or -S(=O)2NH2 (wherein each R #(These are independent, as defined above.)

[0096] The "phosphonate" group is represented by the formula: -P(=O)(O(R # ))2, -P(=O)(OH)2, -OP(=O)(O(R # ))(R # ), or -OP(=O)(OH)(R # ) is a radical (where each R # (These are independent, as defined above.)

[0097] The "phosphine" group is represented by the formula: -P(R # )2 radicals (where each R # (These are independent, as defined above.)

[0098] Where a group described herein (with the exception of alkyl groups) is expressed as "substituted," that group may be substituted with any suitable substituent(s) (one or more). Exemplary examples of substituents include those found in the compounds and embodiments disclosed herein, as well as halogens (chloro, iodo, bromo, or fluoro), alkyl, hydroxyl, alkoxy, alkoxyalkyl, amino, alkylamino, carboxy, nitro, cyano, thiol, thioether, imine, imide, amidine, guanidine, enamine, aminocarbonyl, acylamino, phosphonate, phosphine, thiocarbonyl, sulfinyl, sulfone, sulfonamide, ketone, aldehyde, ester, urea, urethane, oxime, hydroxylamine, alkoxyamine, aralkoxyamine, N-oxide, hydrazine, hydrazide, hydrazone, azide, isocyanate, isothiocyanate, cyanate, thiocyanate, oxygen (=O), B(OH)2, O(alkyl)aminocarbonyl, monocyclic, condensed or These include cycloalkyl compounds that may be non-condensed polycyclic (e.g., cyclopropyl, cyclobutyl, cyclopentyl, or cyclohexyl), or heterocyclil compounds that may be monocyclic, condensed, or non-condensed polycyclic (e.g., pyrrolidyl, piperidyl, piperazinyl, morpholinyl, or thiadinyl), monocyclic, condensed, or non-condensed polycyclic aryl or heteroaryl compounds (e.g., phenyl, naphthyl, pyrrolyl, indolyl, furanil, thiophenyl, imidazolyl, oxazolyl, isoxazolyl, thiazolyl, triazolyl, tetrazolyl, pyrazolyl, pyridinyl, quinolinyl, isoquinolinyl, acridinyl, pyrazinyl, pyridadinyl, pyrimidinyl, benzimidazolyl, benzothiophenyl, or benzofuranil), aryloxys, aralkyloxys, heterocyclyloxys, and heterocyclylalkoxys.

[0099] As used herein, “pharmaceutically acceptable salts” means salts prepared from pharmaceutically acceptable, non-toxic acids or bases, including inorganic acids or bases and organic acids or bases.

[0100] As used herein, unless otherwise indicated, the term “solvate” means a compound or a salt thereof that further contains a stoichiometric or non-stoichiometric amount of solvent bonded by non-covalent intermolecular forces. In one embodiment, the solvate is a hydrate.

[0101] As used herein, unless otherwise indicated, the term “hydrate” means a compound or salt thereof that further contains stoichiometric or nonstoichiometric amounts of water bound by non-covalent intermolecular forces.

[0102] As used herein, unless otherwise indicated, the term “prodrug” means a compound derivative that can be hydrolyzed, oxidized, or otherwise reacted under biological conditions (in vitro or in vivo) to provide an active compound. Examples of prodrugs, but not limited to, are derivatives and metabolites of compounds containing biohydrolyzable sites, such as biohydrolyzable amides, biohydrolyzable esters, biohydrolyzable carbamates, biohydrolyzable carbonates, biohydrolyzable ureids, and biohydrolyzable phosphate analogs. In certain embodiments, a prodrug of a compound having a carboxyl functional group is a lower alkyl ester of a carboxylic acid. Carboxylic acid esters can be formed by esterifying any of the carboxylic acid moieties present on the molecule. Typically, prodrugs are formed by well-known methods (e.g., Burger's Medicinal Chemistry and Drug Discovery 6). th It can be prepared using the methods described in (Donald J. Abraham ed., 2001, Wiley) and Design and Application of Prodrugs (H. Bundgaard ed., 1985, Harwood Academic Publishers GmbH).

[0103] Where used herein, unless otherwise specified, the terms “stereoisomer” or “stereoisomerically pure” mean a stereoisomer of a compound that substantially contains no other stereoisomers of that compound. For example, a stereoisomerically pure compound having one chiral center substantially contains no opposite enantiomer of the compound. A stereoisomerically pure compound having two chiral centers substantially contains no other diastereomers of the compound. Typical stereoisomerically pure compounds contain about 80% by weight of one stereoisomer and less than about 20% by weight of the other stereoisomer of the compound, about 90% by weight of one stereoisomer and less than about 10% by weight of the other stereoisomer of the compound, about 95% by weight of one stereoisomer and less than about 5% by weight of the other stereoisomer of the compound, or about 97% by weight of one stereoisomer and less than about 3% by weight of the other stereoisomer of the compound. Compounds may have chiral centers and may exist as racemates, individual enantiomers or diastereomers, and mixtures thereof. All such isomeric forms, including mixtures thereof, are included in the embodiments disclosed herein. The use of stereoisomerically pure forms of such compounds, as well as mixtures of those forms, are encompassed by the embodiments disclosed herein. For example, mixtures containing equimolar or unequal amounts of enantiomers of a particular compound may be used in the methods and compositions disclosed herein. These isomers can be synthesized asymmetrically or resolved using standard techniques (e.g., chiral columns or chiral resolving agents).For example, see Jacques, J., et al., Enantiomers, Racemates and Resolutions (Wiley Interscience, New York, 1981); Wilen, SH, et al., Tetrahedron 33:2725 (1977); Eliel, EL, Stereochemistry of Carbon Compounds (McGraw Hill, NY, 1962); and Wilen, SH, Tables of Resolving Agents and Optical Resolutions p.268 (EL Eliel, Ed., Univ. of Notre Dame Press, Notre Dame, IN, 1972).

[0104] It should also be noted that the compound may include E and Z isomers or mixtures thereof, as well as cis and trans isomers or mixtures thereof. In certain embodiments, the compound is isolated as either a cis or trans isomer. In other embodiments, the compound is a mixture of cis and trans isomers.

[0105] A "tautomer" refers to an isomeric form of a compound that is in equilibrium with itself. The concentration of the isomers depends on the environment in which the compound is found and can vary depending on whether the compound is a solid or in an organic solution or aqueous solution. For example, in aqueous solution, pyrazole may exhibit the following isomers, which are called tautomers of each other. [ka]

[0106] As will be readily apparent to those skilled in the art, a wide variety of functional groups and other structures may exhibit tautomerism, and all tautomers of a compound are within the scope of this disclosure.

[0107] It should also be noted that compounds may contain unnatural proportions of atomic isotopes in one or more atoms. For example, a compound may contain tritium (3 H), Iodine-125( 125 I), Sulfur-35( 35 S), or carbon-14 ( 14 It may be radioactively labeled with radioactive isotopes such as C, or deuterium ( 2 H), carbon-13 ( 13 C), or nitrogen-15( 15 They may be isotope-enriched, such as N). As used herein, “isotopologs” refers to isotope-enriched compounds. The term “isotopologs” means that an atom has an isotope composition different from that of its natural isotope composition. “Isotope-enriched” may also refer to a compound containing at least one atom in which the atom has an isotope composition different from that of its natural isotope composition. The term “isotopologs” refers to the amount of each isotope present relative to a given atom. Radiolabeled and isotope-enriched compounds are useful as therapeutic agents, e.g., cancer and inflammation treatments; research reagents, e.g., binding assay reagents; and diagnostic agents, e.g., in vivo contrast agents. All isotope variations of the compounds described herein, whether radioactive or not, are intended to be included within the scope of the embodiments provided herein. In some embodiments, isotope-substituted compounds of the compounds are provided, for example, the isototopologs are deuterium, carbon-13, or nitrogen-15 enriched compounds.

[0108] Please note that in the event of any inconsistency between the illustrated structure and its name, the illustrated structure will take precedence.

[0109] In this specification, the term “residue” refers to a chemical portion of a compound that remains after a chemical reaction. For example, the terms “amino acid residue” or “N-alkyl amino acid residue” refer to the product of an amide coupling or peptide coupling of an amino acid or N-alkyl amino acid with a suitable coupling partner, for example, a water molecule being expelled after the amide or peptide coupling of an amino acid or N-alkyl amino acid, resulting in the incorporation of an amino acid residue or N-alkyl amino acid residue into the product.

[0110] As used herein, “sugar,” “sugar group,” or “sugar residue” refers to a carbohydrate portion that may include 3-carbon (triose) units, 4-carbon (tetrose) units, 5-carbon (pentose) units, 6-carbon (hexose) units, 7-carbon (heptose) units, or combinations thereof, and may be a monosaccharide, disaccharide, trisaccharide, tetrasaccharide, pentasaccharide, oligosaccharide, or any other polysaccharide. In some cases, “sugar,” “sugar group,” or “sugar residue” may include furanoses (e.g., ribofuranose, fructofuranose) or pyranoses (e.g., glucopyranose, galactopyranose), or combinations thereof. In some cases, “sugar,” “sugar group,” or “sugar residue” may include aldoses or ketoses, or combinations thereof. Non-limiting examples of monosaccharides include ribose, deoxyribose, xylose, arabinose, glucose, mannose, galactose, and fructose. Non-exclusive examples of disaccharides include sucrose, maltose, lactose, lactulose, and trehalose. Other “sugars,” “sugar groups,” or “sugar residues” include, but are not limited to, polysaccharides and / or oligosaccharides, including amylose, amylopectin, glycogen, inulin, and cellulose. In some cases, the “sugar,” “sugar group,” or “sugar residue” is an amino sugar. In some cases, the “sugar,” “sugar group,” or “sugar residue” is a glucamine residue (1-amino-1-deoxy-D-glucitol) (i.e., glucamide) that attaches to the remainder of the molecule via an amino group, forming an amide bond with the remainder of the molecule.

[0111] Certain groups, subgroups, substituents, and atoms are represented, for example, by a wavy line crossing the bond(s)(single or multiple) to indicate the atom to which that group, subgroup, substituent, or atom is bonded. For example, a phenyl group is represented as a propyl group as shown below: [ka] When replaced by , it has the following structure. [ka]

[0112] Diagrams showing substituents attached to an acyclic group via a bond between two atoms mean, unless otherwise specified, that the substituent may be attached to any atom of the bond through which the substituent bond passes, according to the techniques described herein or techniques known in the art to which this disclosure belongs. Therefore, for example, [ka] teeth [ka] It includes.

[0113] As used herein, “binding agent” refers to any molecule (e.g., an antibody) that is capable of binding with specificity to a given binding partner (e.g., an antigen).

[0114] As used herein, the term “amino acid” refers to an organic compound that contains an amino group (-NH2) and a carboxyl group (-COOH) along with a side chain (R group) specific to each amino acid. Amino acids can be either proteinogenic or non-proteinogenic. “Protogenic” means that an amino acid is one of the 20 naturally occurring amino acids found in proteins. Examples of proteinogenic amino acids include alanine, arginine, asparagine, aspartic acid, cysteine, glutamine, glutamic acid, glycine, histidine, isoleucine, leucine, lysine, methionine, phenylalanine, proline, serine, threonine, tryptophan, tyrosine, and valine. “Non-proteinogenic” means that the amino acid is not naturally present in proteins or is not directly produced by cellular mechanisms (e.g., it is a product of post-translational modification). Non-extendable examples of non-proteinogenic amino acids include gamma-aminobutyric acid (GABA), taurine (2-aminoethanesulfonic acid), theanine (L-γ-glutamylethylamide), hydroxyproline, beta-alanine, ornithine, and citrulline.

[0115] As used herein, “peptide” is defined in its broadest sense across its various grammatical forms and refers to a compound of two or more subunit amino acids, amino acid analogs, or other peptide mimetic compounds. The subunits may be linked by peptide bonds or by other bonds (e.g., esters, ethers, etc.). As used herein, the term “amino acid” refers to natural and / or unnatural or synthetic amino acids (including glycine and both D and L optical isomers), as well as amino acid analogs and peptide mimetic compounds. When the peptide chain is short (e.g., two, three, or more amino acids), it is generally called an oligopeptide. When the peptide chain is longer, the peptide is usually called a polypeptide or protein. Full-length proteins, analogs, variants, and fragments of these are included in this definition. The term also includes post-expression modifications of polypeptides, such as glycosylation, acetylation, and phosphorylation. Furthermore, because ionizable amino and carboxyl groups are present in the molecule, certain peptides can be obtained as acidic or basic salts, or in a neutral form. Peptides can be obtained directly from source organisms, or they can be produced through recombinant DNA or synthesis.

[0116] The amino acid sequences of antibodies can be numbered using any known numbering scheme, including those described in Kabat et al. ("Kabat" numbering scheme); Al-Lazikani et al., 1997, J. Mol. Biol., 273:927-948 ("Chothia" numbering scheme); MacCallum et al., 1996, J. Mol. Biol. 262:732-745 ("Contact" numbering scheme); Lefranc et al., Dev. Comp. Immunol., 2003, 27:55-77 ("IMGT" numbering scheme); and Honegge and Pluckthun, J. Mol. Biol., 2001, 309:657-70 ("AHo" numbering scheme). Unless otherwise specified, the numbering scheme used herein is the Kabat numbering scheme. However, the selection of a numbering scheme is not intended to imply sequence differences where they do not exist, and those skilled in the art can easily determine the sequence location by examining the amino acid sequences of one or more antibodies. Unless otherwise stated, the "EU numbering scheme" is generally used to refer to residues within the constant region of the antibody heavy chain (as reported, e.g., by Kabat et al. (cited above)).

[0117] As used herein, the term “anti-HER2 antibody” refers to an antibody that selectively binds to the HER2 receptor (e.g., trastuzumab (Herceptin)). In one embodiment, trastuzumab may be prepared and used as described in US6407213 and US5821337. The entire disclosures of these documents are incorporated herein by reference.

[0118] As used herein, the term "ifinatamab" refers to an antibody that selectively binds to the B7H3 receptor. In one embodiment, ifinatamab may be prepared and used as described in US10117952 or WO2022102695. The entire disclosures of these documents are incorporated herein by reference.

[0119] As used herein, the term "cytotoxic activity" refers to activity that reduces or diminishes the cell viability of the cell line being tested.

[0120] In the following claims and prior descriptions, unless the context requires other meanings by explicit expression or necessary implied meaning, the word “comprise” or variations such as “comprises” or “comprising” are used in a comprehensive sense, that is, to specify the presence of the described features, but not to exclude the presence or addition of further features in various embodiments.

[0121] 7.2. Conjugate In some embodiments, the conjugate, or a pharmaceutically acceptable salt thereof, tautomer, solvate, or stereoisomer thereof, comprises a protein bound to at least one payload or payload residue (also referred herein as a drug unit) and to at least one hydrophilic moiety via a covalent linker. The covalent linker is directly or indirectly bound to each of the protein, payload residue, and hydrophilic moiety. In some embodiments, the protein is a binder such as an antibody or its antigen-binding fragment.

[0122] In some embodiments, the protein is directly bound to a covalent linker (e.g., a linker) as described herein. In such cases, the binder is located one binding site away from the covalent linker. The covalent linker may also be directly bound to a payload residue, such that the covalent linker is located one binding site away from the payload residue. The payload may be any payload as described herein. In some embodiments, the covalent linker is further directly bound to a hydrophilic moiety, such that the covalent linker is located one binding site away from the hydrophilic moiety. The hydrophilic moiety may be any hydrophilic moiety (HG) as described herein.

[0123] In some embodiments, the linker is indirectly attached to the covalent linker such that the linker is present through a plurality of attachment positions from the covalent linker. In such cases, the linker is attached to the covalent linker through another moiety. For example, the linker can be attached to a maleimide group that is attached to a polyethylene glycol group that is attached to the covalent linker.

[0124] In some examples, the covalent linker is further indirectly attached to the payload residue such that the covalent linker is present through a plurality of attachment positions from the payload residue. The covalent linker is attached to the payload through another moiety. For example, the covalent linker can be attached to a dipeptide (e.g., but not limited to, Val-Ala or Val-Cit), the dipeptide can be attached to PAB, and PAB can be attached to the payload residue.

[0125] In some embodiments, the covalent linker is indirectly attached to the hydrophilic moiety such that the covalent linker is present through a plurality of attachment positions from the hydrophilic moiety. The covalent linker is attached to the hydrophilic moiety through another moiety.

[0126] 7.2.1. Aspect 1. Provided herein are, for example, ADCs for use in therapy (e.g., cancer therapy).

[0127] One embodiment is an ADC compound of formula (I): [Chemical formula] Or a pharmaceutically acceptable salt, tautomer, solvate, or stereoisomer thereof, wherein BA is a linker selected from a humanized antibody, chimeric antibody, human antibody, or an antigen-binding fragment thereof, U is an arylene, heteroarylene, or a bond, V is a bond or -C≡C-(CH2) n-and, n is an integer between 0 and 10. A is a residue in the stretcher unit, The subscript a' is either 0 or 1. W is a cleaveable unit, The subscript w' is either 0 or 1. Y is a spacer unit, The subscript y' is either 0 or 1. PA is payload residue. The subscript x is between 1 and 15.

[0128] In one embodiment, the subscript x is 1 to 15. In one embodiment, the subscript x is 1 to 12. In one embodiment, the subscript x is 1 to 10. In one embodiment, the subscript x is 2 to 10. In one embodiment, the subscript x is 3 to 10. In one embodiment, the subscript x is 4 to 10. In one embodiment, the subscript x is 4 to 9. In one embodiment, the subscript x is 4 to 8.

[0129] In some embodiments, U and V are not coupled simultaneously.

[0130] In one embodiment, U is a bond. In one embodiment, U is arylene. In one embodiment, U is phenylene. In some embodiments, U is [ka] In one embodiment, U is a heteroarylene. In one embodiment, U is a divalent pyrimidine ring. In one embodiment, U is [ka] In one embodiment, U is [ka] In one embodiment, U is [ka] That is the case.

[0131] In one embodiment, V is a bond. In one embodiment, V is -C≡C-(CH2) n -. In one embodiment, V is -C≡C-(CH2)3-.

[0132] In some embodiments, A is -(CH2) m -C(=O)-, -CH2-C(=O)-NH-(CH2) m -C(=O)-, -(CH2CH2O) m -CH2CH2-C(=O)-, -CH[-(CH2)] m -COOH]-C(=O)-, -CH2-C(=O)-NH-(CH2) m -C(=O)-NH-(CH2) m -C(=O)-, -C(=O)-(CH2) m -C(=O)-, -NH-(CH2) m -C(=O)-, or -NH-(CH2CH2O) m The equation is -CH2CH2-C(=O)-, where each m independently represents an integer of 1, 2, 3, 4, or 5.

[0133] In some embodiments, W is one of the following equations: [ka] In the formula, HG represents the hydrophilic portion or hydrogen.

[0134] In some embodiments, HG is a saccharide, a phosphate ester, a sulfate ester, a phosphodiester, or a phosphonate.

[0135] In some embodiments, HG is a saccharide selected from β-D-galactose, N-acetyl-P-D-galactosamine, N-acetyl-a-D-galactosamine, N-acetyl-P-D-glucosamine, β-D-glucuronic acid, a-L-iduronic acid, a-D-galactose, a-D-glucose, β-D-glucose, a-D-mannose, β-D-mannose, a-L-fucose, β-D-xylose, neuraminic acid, or any analog or modification thereof, or a sulfate, phosphate, carboxyl, amino, or O-acetyl modification thereof.

[0136] In some embodiments, HG is

Chemical formula

Chemical formula

[0137] In some embodiments, Y is -NH-CH2-O- or -NH-(p-C6H4)-CH2-O-.

[0138] In some embodiments, the ADC compound of formula (I) has one of the following formulas,

Chemical formula

[0139] In some embodiments, the moiety

Chemical formula

Chemical formula

Chemical formula

[0140] In some embodiments, stretcher units (-A-) are present, extending the framework of the covalent linker to increase the distance between the conjugator assembly and the drug unit (payload or payload residue). The conjugator assembly may contain components of formula (I) other than the binder (BA), stretcher unit (A), cleavable unit (W), spacer unit (Y), and payload residue (PA). The conjugator assembly may contain components of formula (II) other than the stretcher unit (A), cleavable unit (W), spacer unit (Y), and payload residue (PA). The conjugator assembly may contain components of formula (III) other than the stretcher unit (A). In various embodiments, the conjugator contains a methylsulfonyl group, a thiadiazole group, U, V, and a carbonyl group of formula (I), (II), and / or (III). A stretcher unit can combine a conjugator assembly with a cleavable unit if one exists, a conjugator assembly with a spacer unit if one does not exist but a spacer unit does, and a conjugator assembly with a drug unit if neither a cleavable unit nor a spacer unit exists. A stretcher unit can be combined with multiple cleavable units, spacer units, and / or drug units. The conjugator assembly, cleavable unit, spacer unit, and drug unit may be the conjugator assembly, cleavable unit, spacer unit, and drug unit described herein, respectively.

[0141] Stretcher units can modify the physicochemical properties of the drug-linker depending on the components of the stretcher unit. In some embodiments, stretcher units can enhance the solubility of the drug-linker and may contain one or more solubility-enhancing groups, such as ionic groups or water-soluble polymers. Water-soluble polymers are soluble in water at room temperature and may contain other polymers, such as polyethyleneimines, in addition to poly(ethylene) glycol groups.

[0142] A stretcher unit can contain one or more stretcher groups. An example of a stretcher group is, for example, -C 1~10 Alkylene-C(O)-, -C 1~10 Alkylene-C(O)-NH-C 1~10 Alkylene -C(O)-, -(CH2CH2O) u -CH2CH2-C(O)-, -CH[-C 1~10 Alkylene-COOH]-C(O)-,-C 1~10 Alkylene-C(O)-NH-C 1~10 Alkylene-C(O)-NH-C 1~10 Alkylene-C(O)-,-C(O)-C 1~10 Alkylene-C(O)-,-NH-C 1~10 Alkylene -C(=O)-, -NH-(CH2CH2O) u -CH2CH2-C(=O)-, -NH-C 1~10 Alkylene-,-NH-C 1~10 Alkylene-NH-C(O)-C 1~10 Alkylene-,-NH-C 1~10 Alkylene-C(O)-NH-C 1~10 Alkylene-,-NH-(CH2CH2O) u -, -NH-(CH2CH2O) u -CH2-, -NH- (CH2CH2NH) u -(CH2) u -NH-(CH2CH2NH) u -(CH2) u -NH-C(O)-(CH2) uExamples include -NH-(C3-C8 carbocyclo)-, -NH-(arylene)-, and -NH-(C3-C8 heterocyclo)-, where each u is independently between 1 and 10.

[0143] In some embodiments, the cleavable unit (-W w’ -) exists, and if a spacer unit exists, it can connect the conjugator assembly to the spacer unit, and if a spacer unit does not exist, it can connect the conjugator assembly to the drug unit. The connection from the conjugator assembly to the spacer unit or drug unit can be performed directly from the conjugator assembly if a stretcher unit does not exist, or via the stretcher unit if a stretcher unit exists.

[0144] In some embodiments, the cleavable unit is directly conjugated at one end to a conjugator assembly and at the other end to a drug unit. In some embodiments, the cleavable unit is directly conjugated at one end to a stretcher unit and at the other end to a drug unit. In further embodiments, the cleavable unit is directly conjugated at one end to a stretcher unit and at the other end to a spacer unit. In further embodiments, the cleavable unit is directly conjugated at one end to a conjugator assembly and at the other end to a spacer unit. In some embodiments, the stretcher unit and / or spacer unit may not be present.

[0145] A cleavable unit can form a cleavable bond with a drug unit (PA) or a spacer unit. Examples of reactive groups for forming a cleavable bond include a sulfhydryl group for forming a disulfide bond, an aldehyde, ketone, or hydrazine group for forming a hydrazone bond, a carboxyl or amino group for forming a peptide bond, and a carboxyl or hydroxyl group for forming an ester bond.

[0146] Examples of cleavable units include disulfide-containing linkers that can be cleaved through disulfide exchange, acid-unstable linkers at acidic pH, or linkers that can be cleaved by enzymes such as hydrolase, peptidase, esterase, and gluconidase. A cleavable unit may contain one or more cleavage sites.

[0147] In some embodiments, the cleavable unit contains one or more (e.g., 1 to 12) amino acids. Examples of cleavable units include monopeptides, dipeptides, tripeptides, tetrapeptides, pentapeptides, hexapeptides, heptapeptides, octapeptides, nonapeptides, decapeptides, undecapeptides, or dodecapeptides.

[0148] Each amino acid may be a natural or non-natural amino acid, and / or a D-isomer or L-isomer thereof, as long as a cleavable bond is available. In some embodiments, the cleavable unit consists only of natural amino acids. Each amino acid may be a proteinogenic or non-proteinogenic amino acid.

[0149] In some embodiments, each amino acid is independently selected from the group consisting of alanine, arginine, aspartic acid, asparagine, histidine, glycine, glutamic acid, glutamine, phenylalanine, lysine, leucine, serine, tyrosine, threonine, isoleucine, proline, tryptophan, valine, cysteine, methionine, selenocysteine, ornithine, penicillamine, β-alanine, aminoalkanoic acid, aminoalkynic acid, aminoalkanediic acid, aminobenzoic acid, aminoheterocycloalkanoic acid, heterocyclocarboxylic acid, citrulline, statin, diaminoalkanoic acid, and derivatives thereof. In some embodiments, each amino acid is independently selected from the group consisting of alanine, arginine, aspartic acid, asparagine, histidine, glycine, glutamic acid, glutamine, phenylalanine, lysine, leucine, serine, tyrosine, threonine, isoleucine, proline, tryptophan, valine, cysteine, methionine, and selenocysteine.

[0150] In some embodiments, each amino acid is independently selected from the following L-isomers of native amino acids: alanine, arginine, aspartic acid, asparagine, histidine, glycine, glutamic acid, glutamine, phenylalanine, lysine, leucine, serine, tyrosine, threonine, isoleucine, tryptophan, and valine. In some embodiments, each amino acid is the following D-isomer of native amino acids: alanine, arginine, aspartic acid, asparagine, histidine, glycine, glutamic acid, glutamine, phenylalanine, lysine, leucine, serine, tyrosine, threonine, isoleucine, tryptophan, and valine.

[0151] In various embodiments, the cleavable unit is the dipeptide -Val-Cit-, -Phe-Lys-, or -Val-Ala.

[0152] In some embodiments, the cleavable unit comprises one or two terminal amino acids, which are bonded to drug units and / or spacer units via functional groups present at the terminal amino acids, for example, their carboxylic acid or amino terminus.

[0153] In some embodiments, the bond between the cleavable unit and the drug unit can be enzymatically cleaved by one or more enzymes, including tumor-associated proteases, to release the drug unit (-PA), which is protonated in vivo upon release to obtain the drug (PA).

[0154] Useful cleavable units can be designed to optimize selectivity for enzymatic cleavage by specific enzymes (e.g., tumor-associated proteases). In one embodiment, the bond between the cleavable unit and the drug unit or spacer unit is a bond whose cleavage is catalyzed by cathepsin B, C, and / or D, or plasmin proteases.

[0155] In some embodiments, the spacer unit (-Y y’-) exists and extends the framework of the covalent linker. Spacer units can link cleavable units and drug units, or stretcher units and drug units, or conjugator assemblies and drug units. Spacer units may contain one or more self-sacrificing or non-self-sacrificing groups. In some embodiments, spacer units contain one or more self-sacrificing groups. In this context, the term “self-sacrificing group” refers to a bifunctional chemical moiety that can covalently bond two separated chemical moieties together to form a normally stable tripartite molecule. The self-sacrificing group spontaneously separates from the second chemical moiety when the bond with the first moiety is broken. In other embodiments, spacer units are not self-sacrificing. In such embodiments, some or all of the spacer unit remains bonded to the drug unit.

[0156] In some embodiments, -Y y’ - is a self-sacrificing group, which is bonded to the cleavable unit via the methylene carbon atom of the self-sacrificing group, and directly to the drug unit via a carbonate group, carbamate group, or ether group.

[0157] In some embodiments, -Y y’ The hyphen (-) represents a p-aminobenzyl alcohol (PAB) unit (e.g., -NH-(C6H4)-CH2-OC(=O)-). The phenylene portion of the PAB unit is optionally substituted with -C1-C8 alkyl, -O-(C1~C8 alkyl), -halogen, -nitro, or -cyano.

[0158] In some embodiments, -Y y’ - is -NH-C 1~10 Alkylene-O- or -NH-(p-C6H4)-C 1~10 It is alkylene-O-.

[0159] In another embodiment, -Y y’ - represents a carbonate group.

[0160] Other examples of self-sacrificing groups include, but are not limited to, aromatic compounds that are electronically similar to the PAB unit, such as 2-aminoimidazole-5-methanol derivatives (see, e.g., Hay et al., 1999, Bioorg. Med. Chem. Lett. 9:2237) and ortho- or para-aminobenzyl acetals. Suitable spacer units include those that undergo cyclization upon amide bond hydrolysis, such as substituted and unsubstituted 4-aminobutyric acid amides (see, e.g., Rodrigues et al., 1995, Chemistry Biology 2:223), appropriately substituted bicyclo[2.2.1] and bicyclo[2.2.2] ring systems (see, e.g., Storm et al., 1972, J. Amer. Chem. Soc. 94:5815), and 2-aminophenylpropionic acid amides (see, e.g., Amsberry et al., 1990, J. Org. Chem. 55:5867). Removal of amine-containing drugs substituted at the α-position of glycine (see, e.g., Kingsbury et al., 1984, J. Med. Chem. 27:1447) is also a suitable example of a self-sacrificing group.

[0161] Other suitable spacer units are disclosed in U.S. Publication No. 2005-0238649, which is incorporated herein by reference.

[0162] Stretcher units, cleavable units, and spacer units suitable for use with the linkers, platforms, and ADCs of this disclosure are described in WO2004 / 010957, WO2007 / 038658, WO2005 / 112919, U.S. Patents 6,214,345, 7,659,241, 7,498,298, 7,968,687, and 8,163,888, and U.S. Publications 2009-0111756, 2009-0018086, and 2009-0274713. Each of these is incorporated herein by reference in whole for any purpose. Binder

[0163] This specification provides, for example, binders (BA or Ab) for use in ADCs as described herein.

[0164] The compound of formula (I) may include any of the BAs described herein.

[0165] In some embodiments, BA is an antibody or its antigen-binding fragment, such as a humanized antibody, a chimeric antibody, or a human antibody, or an antigen-binding fragment thereof.

[0166] In some embodiments, the antibody or its antigen-binding fragment specifically binds to human B7H3. In some embodiments, the antibody or its antigen-binding fragment is ifinatamab.

[0167] In some embodiments, the antibody or its antigen-binding fragment specifically binds to HER2. In some embodiments, the antibody or its antigen-binding fragment is trastuzumab.

[0168] In some embodiments, the antibody or its antigen-binding fragment specifically binds to CLL1. In some embodiments, the antibody or its antigen-binding fragment is 6E7.

[0169] In some embodiments, the antibody or its antigen-binding fragment is a monoclonal antibody, a chimeric antibody, a humanized antibody, a human-modified antibody, a single-chain antibody (scFv), a Fab fragment, a Fab' fragment, or an F(ab')2 fragment. payload

[0170] This specification provides payloads (PAs) for use, for example, in the platforms and / or ADCs described herein.

[0171] The compound of formula (I) may include any PA described herein.

[0172] In some embodiments, each PA is independently a cytotoxic agent.

[0173] In some embodiments, each PA is independently selected from the group consisting of DXd, 7-ethyl-10-hydroxy-camptothecin (SN-38), and monomethyl auristatin E (MMAE).

[0174] In some embodiments, each PA independently comprises a compound of formula (VI): [ka] And, R 9 and R 10 Each of these can independently be hydrogen, halogen, or substituted or unsubstituted C 1~4 It is alkyl.

[0175] In some embodiments, each PA independently [ka] That is the case.

[0176] 7.2.2. Appearance 2. In some embodiments, the ADC compound is represented by one of the following formulas, or by a pharmaceutically acceptable salt, solvate, and / or stereoisomer. [Table 16] In the formula, Ab is a humanized, chimeric, or human antibody, or an antigen-binding fragment thereof, and the subscript x is between 1 and 15. Alternative values ​​for Ab are as described herein (for example, with respect to Embodiment 1). Alternative values ​​for the variable subscript x are as described herein (for example, with respect to the compound of formula (I)).

[0177] 7.2.3. Appearance 3. Furthermore, this specification also provides a platform for use in the preparation of, for example, ADC (for example, the ADC described herein).

[0178] In some embodiments, the platform is a linker-payload compound of formula (II): [ka] or a pharmaceutically acceptable salt, tautomer, solvate, or stereoisomer thereof, in the formula, U is an arylene, heteroarylene, or bond. V represents a bond or -C≡C-(CH2) n -and, n is an integer between 0 and 10. A is a stretcher unit residue, The subscript a' is either 0 or 1. W is a cleaveable unit, The subscript w' is either 0 or 1. Y is a spacer unit, The subscript y' is either 0 or 1. PA is a payload residue.

[0179] In some embodiments, U and V are not coupled simultaneously.

[0180] In one embodiment, U is a bond. In one embodiment, U is arylene. In one embodiment, U is phenylene. In some embodiments, U is [ka] In one embodiment, U is a heteroarylene. In one embodiment, U is a divalent pyrimidine ring. In one embodiment, U is [ka] In one embodiment, U is [ka] In one embodiment, U is [ka] That is the case.

[0181] In one embodiment, V is a bond. In one embodiment, V is -C≡C-(CH2) n -. In one embodiment, V is -C≡C-(CH2)3-.

[0182] In some embodiments, A is -(CH2) m -C(=O)-, -CH2-C(=O)-NH-(CH2) m -C(=O)-, -(CH2CH2O) m -CH2CH2-C(=O)-, -CH[-(CH2)] m -COOH]-C(=O)-, -CH2-C(=O)-NH-(CH2) m -C(=O)-NH-(CH2) m -C(=O)-, -C(=O)-(CH2) m -C(=O)-, -NH-(CH2) m -C(=O)-, or -NH-(CH2CH2O) m The equation is -CH2CH2-C(=O)-, where each m independently represents an integer of 1, 2, 3, 4, or 5.

[0183] In some embodiments, W is one of the following equations: [ka] In the formula, HG represents the hydrophilic portion or hydrogen.

[0184] In some embodiments, HG is a saccharide, a phosphate ester, a sulfate ester, a phosphodiester, or a phosphonate.

[0185] In some embodiments, HG is a saccharide selected from β-D-galactose, N-acetyl-PD-galactosamine, N-acetyl-aD-galactosamine, N-acetyl-PD-glucosamine, β-D-glucuronic acid, aL-iduronic acid, aD-galactose, aD-glucose, β-D-glucose, aD-mannose, β-D-mannose, aL-fucose, β-D-xylose, neuraminic acid, or any analog or modification thereof, or sulfate, phosphate, carboxyl, amino, or O-acetyl modifications thereof.

[0186] In some embodiments, HG is [ka] or these stereoisomers. In some embodiments, HG is [ka] That is the case.

[0187] In some embodiments, Y is -NH-CH2-O- or -NH-(p-C6H4)-CH2-O-.

[0188] In some embodiments, the compound has one of the following formulas: [ka] In the formula, the values ​​of the variables (e.g., PA, A, W, Y, a', w', y', x) are as described above.

[0189] In some embodiments, part [ka] This is one of the following equations: [ka] [ka] In the formula, PA is as described above.

[0190] In some embodiments, the compound is [ka] or a pharmaceutically acceptable salt or solvate thereof.

[0191] 7.2.4. Appearance 4. Furthermore, this specification also provides covalent linkers for use, for example, in the platforms and / or ADCs described herein.

[0192] In some embodiments (for example, an embodiment of a linker for use in a platform), the linker is a compound of formula (III): [ka] or a pharmaceutically acceptable salt, tautomer, or solvate thereof, in the formula, U is an arylene, heteroarylene, or bond. V represents a bond or -C≡C-(CH2) n -and, n is an integer between 0 and 10. A is a stretcher unit, The subscript a' is either 0 or 1.

[0193] In some embodiments, U and V are not coupled simultaneously.

[0194] In one embodiment, U is a bond. In one embodiment, U is arylene. In one embodiment, U is phenylene. In some embodiments, U is [ka] In one embodiment, U is a heteroarylene. In one embodiment, U is a divalent pyrimidine ring. In one embodiment, U is [ka] In one embodiment, U is [ka] In one embodiment, U is [ka] That is the case.

[0195] In one embodiment, V is a bond. In one embodiment, V is -C≡C-(CH2) n -. In one embodiment, V is -C≡C-(CH2)3-.

[0196] In some embodiments, A is a bond, -OH, -CH3, -N(CH3)2, -(CH2) m -C(=O)R 7 -CH2-C(=O)-NH-(CH2) m -C(=O)R 7 -(CH2CH2O) m -CH2CH2-C(=O)R 7 , -CH[-(CH2) m -COOH]-C(=O)R 7 -CH2-C(=O)-NH-(CH2) m -C(=O)-NH-(CH2) m -C(=O)R 7 -C(=O)-(CH2) m -C(=O)R 7 -NH-(CH2) m -C(=O)R 7 , or -NH-(CH2CH2O) m -CH2CH2-C(=O)R 7 And each m is independently 1, 2, 3, 4, or 5, R 7 is OH or NR 8a R 8bAnd R 8a and R 8b Each of these independently represents H, substituted or unsubstituted C. 1~4 Alkyl, substituted, or unsubstituted C 3~5 It is cycloalkyl, or R 8a and R 8b These, together with the atoms to which they bond, are either substituted or unsubstituted C 3~5 It forms a cycloalkyl group.

[0197] In some embodiments, R 7 These are OH, NH2, NHCH3, or N(CH3)2.

[0198] 7.2.5. Appearance 5. In some embodiments, the linker compound is one of the following, or a pharmaceutically acceptable salt and / or solvate thereof. [ka]

[0199] 7.3. Method or process for creating a conjugate This specification provides a method for preparing a conjugate by contacting a linker payload compound with a binder (Ab) under conditions suitable for the formation of a bond between the binder and the linker payload compound. The reaction conditions can be any suitable reaction conditions known in the art. The binder may be an antibody, and the bond may form an antibody-drug conjugate.

[0200] Examples of such reactions are shown in the examples described later.

[0201] In some embodiments, a method for preparing a conjugate includes treating or contacting a compound with a binder under coupling conditions. The compound may include a reactive linker bound to at least one payload. This compound may be any linker compound or platform compound disclosed herein.

[0202] 7.4. Pharmaceutical Compositions This specification also provides compositions (including pharmaceutical compositions) comprising the ADC described herein. In some embodiments, the composition (e.g., a pharmaceutical composition) further comprises pharmaceutically acceptable excipients.

[0203] Pharmaceutical compositions according to this disclosure can be prepared in the form of lyophilized formulations or aqueous solutions by mixing an antibody-drug conjugate of a desired degree of purity with one or more optionally selected pharmaceutically acceptable carriers (Remington's Pharmaceutical Sciences 16th edition, Osol, A. Ed. (1980)). Generally, pharmaceutically acceptable carriers are non-toxic to the recipient at the doses and concentrations used, and such carriers include, but are not limited to, buffers, e.g., phosphates, citrates, and other organic acids; antioxidants (including ascorbic acid and methionine); preservatives (e.g., octadecyldimethylbenzylammonium chloride; hexamethonium chloride; benzalkonium chloride; benzethonium chloride; phenol, butyl, or benzyl alcohol; alkylparabens, e.g., methyl or propylparaben; catechol; resorcinol; cyclohexanol; 3-pentanol; and m-cresol); and low molecular weight (about 1 Polypeptides (less than 0 residues); proteins, e.g., serum albumin, gelatin, or immunoglobulin; hydrophilic polymers, e.g., polyvinylpyrrolidone; amino acids, e.g., glycine, glutamine, asparagine, histidine, arginine, or lysine; monosaccharides, disaccharides, and other carbohydrates (including glucose, mannose, or dextrin); chelating agents, e.g., EDTA; sugars (e.g., sucrose, mannitol, trehalose, or sorbitol); salt-forming counterions, e.g., sodium; metal complexes (e.g., Zn-protein complexes); and / or nonionic surfactants, e.g., polyethylene glycol (PEG). Exemplary pharmaceutically acceptable carriers as used herein include interstitial drug dispersants such as soluble neutral active hyaluronidase glycoproteins (sHASEGP), e.g., human soluble PH-20 hyaluronidase glycoproteins such as rHuPH20 (HYLENEX®; Baxter International, Inc.). Certain exemplary sHASEGPs and uses, including rHuPH20, are described in U.S. Patent Nos. 7,871,607 and 2006 / 0104968.In one embodiment, sHASEGP is combined with one or more glycosaminoglycans (e.g., chondroitinase).

[0204] Examples of lyophilized formulations are described in U.S. Patent No. 6,267,958. Aqueous formulations include those described in U.S. Patent No. 6,171,586 and WO2006 / 044908, the latter of which includes histidine-acetate buffer.

[0205] A sustained-release preparation may be prepared. A suitable example of a sustained-release preparation is a semipermeable matrix of a solid hydrophobic polymer containing an antibody-drug conjugate, the matrix of which may take the form of a molded article (e.g., a film or microcapsule).

[0206] Preparations used for in vivo administration are generally sterile. Sterilization can be easily achieved, for example, by filtration using a sterile filtration membrane.

[0207] 7.5.How to use In some embodiments, this specification provides a method for treating a disease or disorder (e.g., cancer) in a person in need (e.g., a patient), which includes administering an effective amount of an ADC disclosed herein to the patient.

[0208] The antibody-drug conjugates disclosed herein may be administered by any preferred means (including parenteral, intrapulmonary, intranasal, and intrafocal administration as required for local treatment). Parenteral administration may include intramuscular, intravenous, intra-arterial, intraperitoneal, or subcutaneous administration. Administration may be carried out by any preferred route, for example, by injection (e.g., intravenous or subcutaneous injection), depending in part whether the administration is short-term or chronic. Various administration schedules (including, but not limited to, single or multiple doses, bolus administration, and pulse infusion at various time points) are intended.

[0209] The antibody-drug conjugates of this disclosure can be formulated, administered, and given in a manner consistent with good medical practice. Factors to be considered in this context include the specific disorder being treated, the specific mammalian species being treated, the clinical condition of the individual patient, the cause of the disorder, the site of drug delivery, the method of administration, the scheduling of administration, and other factors known to the healthcare provider. [Examples]

[0210] 8. Examples The following examples are illustrative and should not be considered limiting in any way. Unless otherwise specified, the experimental methods in the following examples are conventional. Unless otherwise specified, reagents and materials are commercially available. All solvents and chemicals used are analytical grade or of chemical purity. Solvents were redistilled before use. Anhydrous solvents were prepared according to standard or reference methods. Silica gel for column chromatography (100-200 mesh) and silica gel for thin-layer chromatography (TLC) (GF254) are commercially available from Tsingdao Haiyang Chemical Co., Ltd. or Yantai Chemical Co., Ltd. in China, and both were eluted with petroleum ether (60-90°C) / ethyl acetate (v / v) and visualized with iodine or molybd phosphoric acid solution in ethanol unless otherwise specified. All extraction solvents were dried with anhydrous Na2SO4 unless otherwise specified. 1¹H NMR spectra were recorded using a Bruck-400, Varian 400MR nuclear magnetic resonance spectrometer (using TMS (tetramethylsilane) as an internal standard). Coupling constants are shown in Hertz. Peaks are reported as singlelines (s), doublelines (d), triplelines (t), quadruplines (q), quintuplines (p), hexatlines (h), heptuplines (hept), multilines (m), or combinations thereof. br stands for broad. LC / MS data were recorded using an Agilent 1100, 1200 high-performance liquid chromatography-ion trap mass spectrometer (LC-MSD trap) equipped with a diode array detector (DAD) detecting at 214 nm and 254 nm and an ion trap (ESI source). All compound names except for reagents were generated using ChemDraw® 18.0.

[0211] For the sake of brevity, this specification uses certain abbreviations. One example is the one-letter abbreviations for amino acids. The amino acids and their corresponding three-letter and one-letter abbreviations are as follows: [Table 17]

[0212] In the following examples, the following abbreviations will be used. [Table 18]

[0213] UPLC analysis method Method A: Mobile phase A: 0.1% FA in water, B: MeCN, Gradient: 10% B maintained for 0.2 minutes, 10%~95% B for 5.8 minutes, 95% B maintained for 0.5 minutes, Flow rate: 0.6 mL / min, Column: ACQUITY UPLC(registered trademark) BEH C18 1.7 μm.

[0214] Method B: Mobile phase A: 0.1% FA in water, B: MeCN, Gradient: 10% B maintained for 0.5 minutes, 10%~90% B for 2.5 minutes, 90% B maintained for 0.2 minutes, Flow rate: 0.6 mL / min, Column: ACQUITY UPLC(registered trademark) BEH C18 1.7 μm

[0215] Method C: Mobile phase A: 0.1% FA in water, B: MeCN, Gradient: 10% B maintained for 0.2 minutes, 10%~90% B for 1.3 minutes, 90% B maintained for 0.3 minutes, Flow rate: 0.6 mL / min, Column: ACQUITY UPLC(registered trademark) BEH C18 1.7 μm

[0216] Example 1-1 [ka] Step 1: N,N-dimethylhexa-5-inamide (1-1b)

[0217] To a solution of compound 1-1a (500 mg, 4.46 mmol) in DCM (10 mL), dimethylamine hydrochloride (545 mg, 6.69 mmol), EDCI (1110 mg, 5.8 mmol), HOBt (783 mg, 5.8 mmol), and Et3N (1350 mg, 13.4 mmol) were added. The mixture was stirred at room temperature for 16 hours. The mixture was diluted with EA (200 mL) and washed with 1N HCl (50 mL x 3), saturated NaHCO3 (50 mL x 3), and brine (50 mL x 3). The organic layer was dried over anhydrous Na2SO4, filtered, and concentrated to obtain crude product 1-1b (458 mg, crude) as a pale yellow solid. MS (ESI) m / z: 140.2 [M+H] + .

[0218] Step 2: N,N-dimethyl-6-(2-(methylthio)pyrimidine-5-yl)hexa-5-inamide(1-1d)

[0219] Compound 1-1b (100 mg, 0.72 mmol) was dissolved in DMF (4 mL) and compound 1-1c (162 mg, 0.79 mmol), CuI (14 mg, 0.072 mmol), Pd(PPh3)2Cl2 (36 mg, 0.072 mmol), and Et3N (1 mL) were added. The mixture was stirred at 90°C for 6 hours under an N2 atmosphere. The mixture was filtered through a Celite pad, diluted with EA (100 mL), and washed with brine (50 mL x 4). The organic layer was dried over anhydrous Na2SO4, filtered, and concentrated. The crude product was purified by flash column chromatography (elution at PE:EA = 0-40%). Compound 1-1d (115 mg, yield 60.8%) was obtained as a pale yellow solid. MS (ESI) m / z: 264.1 [M+H] + .

[0220] Step 3: N,N-dimethyl-6-(2-(methylsulfonyl)pyrimidine-5-yl)hexa-5-inamide(1-1)

[0221] m-CPBA (262 mg, 1.52 mmol) was added to a solution of compound 1-1d (100 mg, 0.38 mmol) in DCM (4 mL). The mixture was stirred at room temperature for 30 minutes. The mixture was diluted with EA (150 mL) and washed with saturated NaHCO3 (100 mL) and brine (100 mL × 3). The organic layer was dried over anhydrous Na2SO4, filtered, concentrated, and the residue was purified by preparative HPLC (Method: Column: XBridge Prep C18 OBD 5 μm 19 × 250 mm; Mobile phase: A - Water (0.1% TFA): B - Acetonitrile; Flow rate: 20 mL / min).

[0222] Compound 1-1 (45.8 mg, 99% purity) was obtained as a white solid. MS (ESI) m / z: 296.2 [M+H] + .

[0223] Examples 1-2 [ka] Step 1: 3-bromo-5-(methylthio)-1,2,4-thiadiazole (1-2b)

[0224] Compound 1-2a (5 g, 29.4 mmol) was dissolved in dichloromethane (100 mL), to which Br2 (5.2 g, 32.9 mmol) was added dropwise at 0°C. After addition, the resulting mixture was stirred overnight at room temperature. Excess Na2SO3 and water (50 mL) were added to the reaction mixture to decompose the excess Br2. The mixture was separated, and the separated organic layer was washed with brine (50 mL x 3), dried over Na2SO4, and concentrated under vacuum. The residue was purified by flash column chromatography (elution at PE:EA = 0%~40%) to obtain compound 1-2b (4.2 g, yield 68.2%) as a pale yellow solid. MS (ESI) m / z: 212.9 [M+H] + .

[0225] Step 2: N,N-dimethyl-4-(5-(methylthio)-1,2,4-thiadiazole-3-yl)benzamide (1-2d)

[0226] Compounds 1-2c (110 mg, 0.57 mmol), K3PO4 (201 mg, 0.95 mmol), and Pd(PPh3)4 (36 mg, 0.072 mmol) were added to a solution of compound 1-2b (100 mg, 0.47 mmol) in DMF (4 mL) and H2O (1 mL). The mixture was stirred at 90°C for 6 hours under an N2 atmosphere. The mixture was filtered through a Celite pad, diluted with EA (100 mL), and washed with brine (50 mL x 4). The organic layer was dried over anhydrous Na2SO4, filtered, and concentrated. The crude product was purified by flash column chromatography (elution at PE:EA = 0-40%). Compound 1-2d (35 mg, yield 26.4%) was obtained as a white solid. MS (ESI) m / z: 280.2 [M+H] + .

[0227] Step 3: N,N-dimethyl-4-(5-(methylsulfonyl)-1,2,4-thiadiazole-3-yl)benzamide(1-2)

[0228] m-CPBA (74 mg, 0.43 mmol) was added to a solution of compound 1-2d (30 mg, 0.11 mmol) in DCM (4 mL). The mixture was stirred at room temperature for 3 hours. The mixture was concentrated and purified by preparative HPLC (Method: Column: XBridge Prep C18 OBD 5 μm 19 × 250 mm; Mobile phase: A - Water (0.1% TFA): B - Acetonitrile; Flow rate: 20 mL / min). Compound 1-2 (4.0 mg, yield 12%) was obtained as a white solid.

[0229] MS (ESI) m / z: 312.3 [M+H] + .

[0230] Examples 1-3 [ka] Step 1: tert-butyl(4-(5-(methylthio)-1,2,4-thiadiazole-3-yl)phenyl)carbamate(1-3b)

[0231] Compound 1-2b (150 mg, 0.71 mmol) was dissolved in toluene / EtOH (v:v=7:3, 5 mL), to which 1-3a (202 mg, 0.85 mmol), Na2CO3 (150 mg, 1.42 mmol) in H2O (1 mL), and Pd(PPh3)4 (82 mg, 0.071 mmol) were added. The mixture was heated under N2 reflux for 3 hours. The reaction mixture was then cooled to room temperature and diluted with 5.0 ml of ethyl acetate. The aqueous layer was then separated, the organic layer was washed with saturated NaCl solution, dried over Na2SO4, concentrated, and the residue was purified by flash column chromatography (elution at PE:EA = 0%~40%) to obtain compound 1-3b (75 mg, yield 32.6%).

[0232] MS (ESI) m / z: 324.1 [M+H] + .

[0233] Step 2: 4-(5-(methylthio)-1,2,4-thiadiazole-3-yl)aniline(1-3c)

[0234] TFA (1 mL) was added dropwise to a solution of compound 1-3b (75 mg, 0.23 mmol) in DCM (4 mL). The mixture was stirred at room temperature for 1 hour. The mixture was concentrated and co-evaporated with toluene (3 mL x 3). Compound 1-3b (55 mg, crude) was obtained as a white solid and used in the next step without further purification.

[0235] MS (ESI) m / z: 224.1 [M+H] + .

[0236] Step 3: N-(4-(5-(methylthio)-1,2,4-thiodiazole-3-yl)phenyl)acetamide(1-3d)

[0237] Compound 1-3c (55 mg, 0.25 mmol) was dissolved in THF (4 mL) and acetic anhydride (50.3 mg, 0.49 mmol) and Et3N (50 mg, 0.49 mmol) were added. The mixture was stirred at room temperature for 2 hours. The mixture was filtered, and the filtrate was diluted with EA (50 mL) and washed with saturated NaHCO3 (50 mL x 3) and brine (50 mL x 3), respectively. The organic layer was dried over anhydrous Na2SO4, filtered, and concentrated. The residue was purified by flash column chromatography (elution at PE:EA = 0%~40%). Compound 1-3d (50 mg, yield 76.5%) was obtained as a white solid.

[0238] MS (ESI) m / z: 266.1 [M+H] + .

[0239] Step 4: N-(4-(5-(methylsulfonyl)-1,2,4-thiadiazole-3-yl)phenyl)acetamide(1-3)

[0240] Compound m-CPBA (130 mg, 0.75 mmol) was added to a solution of compound 1-3d (50 mg, 0.19 mmol) in DCM (4 mL). The mixture was stirred at room temperature for 3 hours. The mixture was concentrated and purified by preparative HPLC (Method: Column: XBridge Prep C18 OBD 5 μm 19 × 250 mm; Mobile phase: A - Water (0.1% TFA): B - Acetonitrile; Flow rate: 20 mL / min). Compound 1-3 (12 mg, yield 12.7%) was obtained as a white solid.

[0241] MS (ESI) m / z: 298.1 [M+H] + .

[0242] Examples 1-4 [ka] Step 1: 5-Bromo-N,N-dimethylpyrimidine-2-carboxamide (1-4b)

[0243] To a solution of compound 1-4a (2000 mg, 9.85 mmol) in DMF (40 mL), dimethylamine hydrochloride (1210 mg, 14.78 mmol), HATU (5620 mg, 14.78 mmol), and DIPEA (2550 mg, 19.70 mmol) were added. The mixture was stirred at room temperature under an N2 atmosphere for 3 hours. The mixture was concentrated. The crude product was purified by flash column chromatography (DCM: elution at MeOH = 0-20%). Compound 1-4b (1500 mg, yield 66.1%) was obtained as an off-white solid.

[0244] MS (ESI) m / z: 232.0 [M+H] + .

[0245] Step 2: (2-(dimethylcarbamoyl)pyrimidine-5-yl)boronic acid (1-4c)

[0246] To a solution of compound 1-4b (1.4 g, 6.09 mmol) in 1,4-dioxane (30 mL) and H2O (3 mL), 4,4,4',4',5,5,5',5'-octamethyl-2,2'-bi-(1,3,2-dioxaborolane) (1.85 g, 7.3 mmol) and potassium acetate (1.19 g, 12.2 mmol) were added under a nitrogen atmosphere. The reaction mixture was stirred for 5 minutes, and Pd(dppf)Cl2.DCM (495 mg, 0.60 mmol) was added. The reaction mixture was refluxed for 16 hours. After completely consuming the starting material, the reaction mixture was diluted with water and extracted with ethyl acetate (3 × 100 mL). The combined organic layers were dried over Na2SO4, filtered, and purified by flash column chromatography (eluting at CH2Cl2:MeOH = 0%~20%) to obtain compound 1-4c (1.05 g, yield 88.3%) as a white solid.

[0247] MS (ESI) m / z: 196.0 [M+H] + .

[0248] Step 3: N,N-dimethyl-5-(5-(methylthio)-1,2,4-thiadiazole-3-yl)pyrimidine-2-carboxamide(1-4d)

[0249] Compound 1-2c (150 mg, 0.71 mmol) was dissolved in toluene / EtOH (v:v=7:3, 5 mL), to which 1-2b (166 mg, 0.85 mmol), Na2CO3 (150 mg, 1.42 mmol) in H2O (1 mL), and Pd(PPh3)4 (82 mg, 0.071 mmol) were added. The mixture was heated under N2 reflux for 3 hours. The reaction mixture was then cooled to room temperature and diluted with 5.0 ml of ethyl acetate. The aqueous layer was then separated, the organic layer was washed with saturated NaCl solution, dried over Na2SO4, concentrated, and the residue was purified by flash column chromatography (elution at PE:EA = 0%~40%) to obtain compound 1-4d (45 mg, yield 22.5%).

[0250] MS (ESI) m / z: 282.0 [M+H] + .

[0251] Step 4: N,N-dimethyl-5-(5-(methylsulfonyl)-1,2,4-thiadiazole-3-yl)pyrimidine-2-carboxamide(1-4)

[0252] m-CPBA (123 mg, 0.71 mmol) was added to a solution of compound 1-4d (40 mg, 0.14 mmol) in DCM (3 mL). The mixture was stirred at room temperature for 2 hours.

[0253] The mixture was concentrated and purified by preparative HPLC (Method: Column: XBridge Prep C18 OBD 5um 19×250mm; Mobile phase: A-Water (0.1% TFA):B-Acetonitrile; Flow rate: 20 mL / min). Compounds 1-4 (3.7 mg, yield 9.3%) were obtained as a white solid. MS (ESI) m / z: 314.1 [M+H]+.

[0254] Examples 1-5 [ka] Step 1: (3-bromo-5-(methylsulfonyl)-1,2,4-thiadiazole (1-5a))

[0255] m-CPBA (817.5 mg, 4.737 mmol) was added to a solution of 1-2b (200 mg, 0.947 mmol) in DCM (3 mL). The white suspension was stirred at room temperature for 5 hours. The mixture was quenched with saturated Na2S2O3 (50 mL) and extracted with DCM (30 mL x 2). The combined organic layers were washed with brine (20 mL), dried over Na2SO4, filtered, and the filtrate was concentrated under vacuum to obtain the residue. This was purified by FCC (EA / PE = 0%~20%), and the fraction was concentrated under vacuum to obtain 1-5a (239 mg, yield 95%) as a white solid.

[0256] 1 H NMR (400 MHz, CDCl3) δ 2.76 (s, 1H).

[0257] Step 2: N,N-dimethyl-6-(5-(methylsulfonyl)-1,2,4-thiadiazole-3-yl)hexa-5-inamide(1-5)

[0258] To a mixture of 1-5a (239.0 mg, 0.983 mmol) and 1-5b (150.5 mg, 1.081 mmol) in DMF (4.0 mL), CuI (37.4 mg, 0.197 mmol), Pd(PPh3)Cl2 (69.0 mg, 0.098 mmol), and DIPEA (205 μL, 152.5 mg, 1.18 mmol) were added, and the mixture was degassed three times under a nitrogen atmosphere. The mixture was stirred overnight at room temperature. The mixture was filtered and purified by preparative HPLC (0.1% FA), and the fraction was freeze-dried to obtain 1-5 (13.6 mg, yield 4%) as a yellow oil.

[0259] 1 H NMR (400 MHz, CDCl3) δ 3.38 (s, 3H), 3.01 (s, 3H), 2.95 (s, 3H), 2.60 (t, J = 6.8 Hz, 2H), 2.50 (t, J = 7.2 Hz, 1H), 2.00 (p, J = 7.0 Hz, 2H).

[0260] MS (ESI) m / z: 302.0 [M+H] + .

[0261] 1 H NMR (400 MHz, DMSO) δ 7.09 (s, 2H), 6.99 (t, J = 6.4 Hz, 1H), 4.60 (dd, J = 10.8, 4.0 Hz, 1H), 3.59-3.53 (m, 1H), 3.46-3.38 (m, 1H), 1.32 (s, 9H).

[0262] Example 2-1 [ka] Step 1: N-((((9H-fluoren-9-yl)methoxy)carbonyl)-L-valyl)-O-((2R,3R,4S,5S,6S)-3,4,5-triacetoxy-6-(methoxycarbonyl)tetrahydro-2H-pyran-2-yl)-L-serine(2-1b)

[0263] To a mixture of 2-1a (4.1 g, 4.92 mmol; purchased from MCE) in MeOH (50 mL), THF (100 mL), and DCM (20 mL), wet Pd / C (400 mg, purity 10%) was added. The black suspension was purged three times with an H2 balloon and then stirred at room temperature for 1 hour. The black suspension was filtered through a Celite pad and washed with MeOH (200 mL). The organic layers were combined and concentrated under vacuum to obtain 2-1b (3650 mg, yield 99.8%) as an off-white solid.

[0264] MS (ESI) m / z: 743.6 [M+H] + .

[0265] Step 2: (2R,3R,4S,5S,6S)-2-((S)-2-((S)-2-((((9H-Fluoren-9-yl)methoxy)carbonyl)amino)-3-methylbutanamide)-3-((2-(benzyloxy)-2-oxoethyl)amino)-3-oxopropoxy)-6-(methoxycarbonyl)tetrahydro-2H-pyran-3,4,5-triyltriacetate(2-1d)

[0266] To solutions of 2-1b (3.65 g, 4.92 mmol) and 2-1c (1.66 g, 4.92 mmol) in DMF (50 mL), HATU (1.87 g, 4.92 mmol) and DIPEA (1.59 g, 12.29 mmol) were added. The mixture was stirred at room temperature for 30 minutes. The mixture was purified by FCC (MeOH / DCM = 0-10%), and the fraction was concentrated under vacuum to obtain 2-1d (3.8 g, yield 86.9%) as an off-white foamed solid.

[0267] MS (ESI) m / z: 890.7 [M+H] + .

[0268] Step 3: N-((((9H-fluoren-9-yl)methoxy)carbonyl)-L-valyl)-O-((2R,3R,4S,5S,6S)-3,4,5-triacetoxy-6-(methoxycarbonyl)tetrahydro-2H-pyran-2-yl)-L-cerylglycine(2-1e)

[0269] A mixture of 2-1d (3.8 g, 4.27 mmol) in MeOH (150 mL) and DCM (50 mL) was mixed with wet Pd / C (400 mg, purity 10%). The black suspension was purged three times with an H2 balloon and then stirred at room temperature for 40 minutes. The black suspension was filtered through a Celite pad and washed with MeOH (150 mL). The organic layers were combined and concentrated under vacuum to obtain 2-1e (3.3 g, yield 96.6%) as an off-white solid.

[0270] MS (ESI) m / z: 800.7 [M+H] + .

[0271] Step 4: (2R,3R,4S,5S,6S)-2-((S)-2-((S)-2-((((9H-Fluoren-9-yl)methoxy)carbonyl)amino)-3-methylbutanamide)-3-((Acetoxymethyl)amino)-3-oxopropoxy)-6-(Methoxycarbonyl)tetrahydro-2H-pyran-3,4,5-triyltriacetate(2-1f)

[0272] To a solution of 2-10e (3.3 g, 4.13 mmol) in DMF (30 mL), Pb(OAc)4 (2.74 g, 6.19 mmol), Cu(OAc)2 (74.9 mg, 0.41 mmol), and HOAc (247.8 mg, 4.13 mmol) were added. The resulting dark-colored mixture was purged three times with an N2 balloon and stirred at 65°C for 40 minutes until the mixture turned a deep blue. The mixture was diluted with siRNA (300 mL), washed with brine (100 mL x 3), dried over Na2SO4, filtered, and concentrated under vacuum to obtain the residue. This was purified by FCC (MeOH / DCM = 0-10%), and the fraction was concentrated under vacuum to obtain 2-1f (2.8 g, yield 83.4%) as a pale yellow solid.

[0273] MS (ESI) m / z: 836.6 [M+Na] + .

[0274] Step 5: (2R,3R,4S,5S,6S)-2-((S)-2-((S)-2-((((9H-Fluoren-9-yl)methoxy)carbonyl)amino)-3-methylbutanamide)-3-(((3-(benzyloxy)-2,2-dimethyl-3-oxopropoxy)methyl)amino)-3-oxopropoxy)-6-(methoxycarbonyl)tetrahydro-2H-pyran-3,4,5-triyltriacetate(2-1h)

[0275] A white suspension mixture of 2-1f (300 mg, 0.37 mmol), 2-1g (153.7 mg, 0.74 mmol), and 4Å molecular sieves (200 mg) in anhydrous THF (10 mL) was stirred at room temperature for 10 minutes. Sc(OTf)3 (217.9 mg, 0.44 mmol) was added, and the resulting yellow suspension was stirred at room temperature for 4 hours. The yellow suspension mixture was filtered through a Celite pad and washed with EA. The combined organic layers were washed with saturated NaHCO3 (30 mL) and brine (30 mL), dried over Na2SO4, filtered, and the filtrate was concentrated under vacuum to obtain the residue. This was purified by silica gel column (MeOH / DCM = 0%~5%), and the fraction was concentrated under vacuum to obtain 2-1h (275 mg, yield 77.5%) as a white foamy solid.

[0276] MS (ESI) m / z: 984.8 [M+Na] + .

[0277] Step 6: (5S,8S)-1-(9H-fluoren-9-yl)-5-isopropyl-14,14-dimethyl-3,6,9-trioxo-8-((((2R,3R,4S,5S,6S)-3,4,5-triacetoxy-6-(methoxycarbonyl)tetrahydro-2H-pyran-2-yl)oxy)methyl)-2,12-dioxa-4,7,10-triazapentadecane-15-euic acid(2-1j)

[0278] To a solution of 2-1h (275 mg, 0.29 mmol) in MeOH (10 mL), wet Pd / C (55 mg, 10% purity) was added. The black suspension was purged three times with an H2 balloon and then stirred at room temperature for 2 hours. The mixture was filtered using a syringe head, washed with MeOH (15 mL), and concentrated under vacuum to obtain 2-1j (230 mg, crude) as a white foamy solid.

[0279] MS (ESI) m / z: 894.6 [M+Na] + .

[0280] Step 7: (2R,3R,4S,5S,6S)-2-((S)-2-((S)-2-((((9H-Fluoren-9-yl)methoxy)carbonyl)amino)-3-methylbutanamide)-3-(((3-(((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indolidino[1,2-b]quinoline-1-yl)amino)-2,2-dimethyl-3-oxopropoxy)methyl)amino)-3-oxopropoxy)-6-(methoxycarbonyl)tetrahydro-2H-pyran-3,4,5-triyltriacetate(2-1k)

[0281] DIPEA (102.3 mg, 0.79 mmol) was added to a mixture of 2-10j (230 mg, crude), exatecan mesylate (139.9 mg, 0.26 mmol), and HATU (100.3 mg, 0.26 mmol) in DMF (5 mL). The resulting brown mixture was stirred at room temperature for 1 hour. The mixture was diluted with  (20 mL), washed with brine (20 mL x 3), dried over Na2SO4, filtered, and concentrated under vacuum to obtain the residue. This was purified by FCC (MeOH / DCM = 0%~3%) and concentrated under vacuum to obtain 2-1k (325 mg, yield 95.6%) as an off-white foamy solid.

[0282] MS (ESI) m / z: 1289.9 [M+H] + .

[0283] Step 8: (2S,3S,4S,5R,6R)-6-((S)-2-((S)-2-amino-3-methylbutanamide)-3-(((3-(((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indolidino[1,2-b]quinoline-1-yl)amino)-2,2-dimethyl-3-oxopropoxy)methyl)amino)-3-oxopropoxy)-3,4,5-trihydroxytetrahydro-2H-pyran-2-carboxylic acid(2-1m)

[0284] Et2N (523.2 mg, 5.06 mmol) was added to a solution of 2-1k (325 mg, 0.25 mmol) in DMF (5 mL). The mixture was stirred at room temperature for 20 minutes. After LC-MS indicated completion of the reaction, the mixture was concentrated under vacuum to obtain the crude product. The crude product was dissolved in MeOH (6 mL). K2CO3 (174.7 mg, 1.26 mmol) was added and the mixture was stirred at room temperature for 10 minutes, then H2O (2 mL) was added to the mixture and the mixture was stirred at room temperature for 30 minutes. The mixture was acidified to pH=3 with saturated KHSO4 at 0°C, filtered, and purified by preparative HPLC (0.1% FA). The fraction was lyophilized to obtain 2-1m (140 mg, yield 59.7%) as a pale yellow solid.

[0285] MS (ESI) m / z: 927.4 [M+H] + .

[0286] 1 H NMR (400 MHz, d6-DMSO) δ 9.56 (s, 1H), 8.39 (s, 1H), 8.07 (d, J = 8.4 Hz, 1H), 7.77 (d, J = 11.2 Hz, 1H), 7.31 (s, 1H), 6.52 (s, 1H), 5.54 (dd, J = 13.2, 7.2 Hz, 1H), 5.43 (s, 2H), 5.18 (dd, J = 41.6, 18.8 Hz, 2H), 5.09 - 5.02 (m, 1H), 4.96 (s, 1H), 4.62 (dd, J = 10.0, 6.8 Hz, 1H), 4.56 - 4.44 (m, 2H), 4.19 (d, J = 7.6 Hz, 1H), 3.82 (dd, J = 10.8, 6.8 Hz, 1H), 3.61 (dd, J = 11.6, 6.4 Hz, 2H), 3.17-3.05 (m, 4H), 2.94 (t, J = 8.0 Hz, 1H), 2.39 (s, 3H), 2.11 (dt, J = 21.3, 7.6 Hz, 2H), 2.03 - 1.93 (m, 2H), 1.92 - 1.78 (m, 3H), 1.12 (d, J = 8.0 Hz, 6H), 0.87 (dd, J = 13.0, 6.6 Hz, 9H).

[0287] ステップ9:6-(2-(メチルチオ)ピリミジン-5-イル)ヘキサ-5-イン acid (2-1q)

[0288] To a mixture of 2-1n (2.0 g, 9.751 mmol) and 2-1p (1.31 g, 11.702 mmol) in DMF (30 mL), CuI (185.7 mg, 0.975 mmol), Pd(PPh3)Cl2 (684.4 mg, 0.975 mmol), and TEA (4.1 mL, 2.960 g, 29.254 mmol) were added. The mixture was degassed three times under a nitrogen atmosphere and then stirred at 95°C for 2 hours. The mixture was diluted with  (30 mL), washed with brine (30 mL x 2), dried over Na2SO4, and the filtrate was concentrated under vacuum to obtain the crude product. This was purified by FCC (MeOH / DCM = 0%~3%), and the fraction was concentrated under vacuum to obtain 2-1q (1.6 g, yield approximately 65%) as a yellow solid.

[0289] MS (ESI) m / z: 237.1 [M+H] + .

[0290] Step 10: 6-(2-(methylsulfonyl)pyrimidine-5-yl)hexa-5-ic acid (2-1r)

[0291] To a solution of 2-1q (520 mg, 1.613 mmol) in DCM (10 mL), m-CPBA (1113.4 mg, 6.452 mmol) was added. The resulting yellow suspension was stirred at room temperature for 1.5 hours. The mixture was diluted with EA (50 mL), quenched by adding saturated Na2S2O3 (30 mL), stirred at room temperature for 10 minutes, and extracted with DCM (30 mL x 3). The organic layers were combined, washed with brine (50 mL), dried over Na2SO4, filtered, and concentrated under vacuum to obtain the crude product as a yellow solid. This was purified by preparative HPLC (FA), and the fraction was freeze-dried to obtain 2-1r (78.0 mg, yield 18%) as a white solid.

[0292] MS (ESI) m / z: 269.1 [M+H] + .

[0293] 1H NMR (400 MHz, d6-DMSO) δ 12.17 (s, 1H), 9.12 (s, 2H), 3.41 (s, 3H), 2.60 (t, J = 7.2 Hz, 2H), 2.41 (t, J = 7.2 Hz, 2H), 1.82 (p, J = 6.8 Hz, 2H).

[0294] Step 11: (2S,3S,4S,5R,6R)-6-((S)-3-(((3-(((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indolidino[1,2-b]quinoline-1-yl)amino)-2,2-dimethyl-3-oxopropoxy)methyl)amino)-2-((S)-3-methyl-2-(6-(2-(methylsulfonyl)pyrimidine-5-yl)hexa-5-inamide)butanamide)-3-oxopropoxy)-3,4,5-trihydroxytetrahydro-2H-pyran-2-carboxylic acid(2-1)

[0295] To a solution of 2-1r (5.2 mg, 0.019 mmol) and 2-1m (15.0 mg, 0.016 mmol) in DMF (2 mL), HATU (8.6 mg, 0.024 mmol) and DIPEA (7 μL, 5.2 mg, 0.04 mmol) were added. The mixture was stirred at room temperature for 30 minutes. The mixture was purified by preparative HPLC (FA 0.1%), and the fraction was freeze-dried to obtain 2-1 (7.1 mg, purity 37.3%) as a white solid.

[0296] MS (ESI) m / z: 1177.9 [M+H] + .

[0297] Example 2-2 [ka] Step 1: tert-butyl 3-(2-(methylthio)pyrimidine-5-carboxamide)propanoate (2-2c)

[0298] To a mixture of 2-2 (600 mg, 3.525 mmol) in DMF (15 mL), HATU (1621.9 mg, 4.266 mmol) and DIPEA (1.9 mL, 1503.5 mg, 11.633 mmol) were added. The mixture was stirred at 45°C for 10 minutes, 2-2b (704.6 mg, 3.878 mmol) was added, and the mixture was stirred at 45°C for 40 minutes. The mixture was diluted with  (50 mL), washed with brine (30 mL x 3), dried over Na2SO4, filtered, and concentrated under vacuum to obtain the residue. This was purified by FCC (MeOH / DCM) to obtain 2-1c (1.3 g, crude) as a brown oil.

[0299] MS (ESI) m / z: 298.4 [M+H] + .

[0300] Step 2: tert-butyl 3-(2-(methylsulfonyl)pyrimidine-5-carboxamide)propanoate (2-2d)

[0301] To a solution of 2-2c (500 mg, crude) in DCM (10 mL), m-CPBA (1160.5 mg, 6.725 mmol) was added. The white suspension was stirred at room temperature for 1 hour. The mixture was diluted with EA (50 mL), quenched by adding saturated Na2S2O3 (30 mL), stirred at room temperature for 10 minutes, and extracted with DCM (30 mL x 3). The organic layers were combined, washed with brine (50 mL), dried over Na2SO4, filtered, and concentrated under vacuum to obtain the crude product as a yellow solid. This was purified by FCC (MeOH / DCM), and the fraction was concentrated under vacuum to obtain 2-2d (303 mg, yield approximately 50%) as a white solid.

[0302] MS (ESI) m / z: 330.3 [M+H] + .

[0303] 1H NMR (400 MHz, cdcl3) δ 9.27 (s, 2H), 7.38 (s, 1H), 3.74 (dd, J = 11.6, 6.0Hz, 1H), 3.40 (s, 2H), 2.60 (t, 1H), 1.47 (s, 5H).

[0304] Step 3: 3-(2-(methylsulfonyl)pyrimidine-5-carboxamide)propanoic acid (2-2e)

[0305] The mixture of 2-2d (303.0 mg, 0.92 mmol) in TFA (1 mL) and DCM (2 mL) was stirred at room temperature for 1.5 hours. The mixture was concentrated under vacuum and evaporated three times with toluene to obtain 2-2e (255 mg) as an off-white solid. MS (ESI) m / z: 274.1 [M+H] + .

[0306] Step 4: (2S,3S,4S,5R,6R)-6-((S)-3-(((3-(((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indolidino[1,2-b]quinoline-1-yl)amino)-2,2-dimethyl-3-oxopropoxy)methyl)amino)-2-((S)-3-methyl-2-(3-(2-(methylsulfonyl)pyrimidine-5-carboxamide)propanamide)butanamide)-3-oxopropoxy)-3,4,5-trihydroxytetrahydro-2H-pyran-2-carboxylic acid(2-2)

[0307] To a solution of 2-2e (7.1 mg, 0.026 mmol) and 2-1m (20.0 mg, 0.022 mmol) in DMF (2 mL), HATU (11.5 mg, 0.030 mmol) and DIPEA (9 μL, 7.0 mg, 0.054 mmol) were added. The mixture was stirred at room temperature for 30 minutes. The mixture was purified by preparative HPLC (FA 0.1%), and the fraction was freeze-dried to obtain 2-2 (13.4 mg, purity approximately 52.5%) as a white solid. MS (ESI) m / z: 1182.8 [M+H] + .

[0308] Examples 2-3 [ka] Step 1: Methyl 4-(5-(methylthio)-1,2,4-thiadiazole-3-yl)benzoate (2-3b)

[0309] Compound 1-2b (100 mg, 0.47 mmol) was dissolved in toluene (4 mL) and H2O (1 mL). Compound 2-3a (109.72 mg, 0.568 mmol), K2CO3 (168 mg, 0.947 mmol), and Pd(dppf)Cl2.DCM (34.6 mg, 0.047 mmol) were added. The mixture was stirred at 110°C for 3 hours under an N2 atmosphere. The mixture was filtered through a Celite pad, diluted with EA (100 mL), and washed with brine (50 mL x 4). The organic layer was dried over anhydrous Na2SO4, filtered, and concentrated. The crude product was purified by flash column chromatography (elution at PE:EA = 0-40%). Compound 2-3b (56 mg, yield 44.4%) was obtained as an off-white solid.

[0310] MS (ESI) m / z: 267.1 [M+H] + .

[0311] Step 2: 4-(5-(methylthio)-1,2,4-thiadiazole-3-yl)benzoic acid (2-3c)

[0312] To a solution of compound 2-3b (54 mg, 0.20 mmol) in MeOH (3 mL) and H2O (1 mL), LiOH (17 mg, 0.41 mmol) was added. The mixture was stirred at room temperature for 2 hours. The mixture was adjusted to pH 7 and purified by preparative HPLC (FA conditions) to obtain compound 2-3c (36 mg, yield 70.3%) as a white solid.

[0313] MS (ESI) m / z: 253.1 [M+H] + .

[0314] Step 3: 4-(5-(methylsulfonyl)-1,2,4-thiadiazole-3-yl)benzoic acid (2-3d)

[0315] m-CPBA (96 mg, 0.55 mmol) was added to a solution of compound 2-3c (35 mg, 0.14 mmol) in DCM (3 mL) and THF (3 mL). The mixture was stirred at room temperature for 16 hours. The mixture was concentrated and purified by preparative HPLC (Method: Column: XBridge Prep C18 OBD 5 μm 19 × 250 mm; Mobile phase: A - Water (0.1% TFA): B - Acetonitrile; Flow rate: 20 mL / min). Compound 2-3d (12 mg, 99% purity) was obtained as a white solid.

[0316] MS (ESI) m / z: 284.8 [M+H] + .

[0317] Step 4: (2S,3S,4S,5R,6R)-6-((S)-3-(((3-(((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indolidino[1,2-b]quinoline-1-yl)amino)-2,2-dimethyl-3-oxopropoxy)methyl)amino)-2-((S)-3-methyl-2-(4-(5-(methylsulfonyl)-1,2,4-thiadiazole-3-yl)benzamide)butanamide)-3-oxopropoxy)-3,4,5-trihydroxytetrahydro-2H-pyran-2-carboxylic acid(2-3)

[0318] Compound 2-3d (7.36 mg, 0.026 mmol) was dissolved in DMF (2 mL), to which HATU (9.02 mg, 0.024 mmol) and DIPEA (5.58 mg, 0.043 mmol) were added. The mixture was stirred at room temperature for 30 minutes. Compound 2-1m (20 mg, 0.022 mmol) was added to the mixture and stirred at room temperature for 15 minutes. The reaction product was purified by preparative HPLC (method: column: XBridge Prep C18 OBD 5um 19 × 250 mm; mobile phase: A - water (0.1% TFA): B - acetonitrile; flow rate: 20 mL / min) to obtain compound 2-3 (7.6 mg, yield 29.5%) as a white solid.

[0319] MS (ESI) m / z: 1193.5 [M+H] + .

[0320] Examples 2-4 [ka] Step 1: (2S,3S,4S,5R,6R)-6-((S)-3-(((3-(((1S,10S)-10-ethyl-10-hydroxy-11,14-dioxo-2,3,10,11,14,16-hexahydro-1H,13H-benzo[de][1,3]dioxolo[4,5-g]pyrano[3',4':6,7]indolidino[1,2-b]quino Phosphate-1-yl)amino)-2,2-dimethyl-3-oxopropoxy)methyl)amino)-2-((S)-3-methyl-2-(4-(5-(methylsulfonyl)-1,2,4-thiadiazole-3-yl)benzamide)butanamide)-3-oxopropoxy)-3,4,5-trihydroxytetrahydro-2H-pyran-2-carboxylic acid(2-4)

[0321] To a solution of 2-3d (5.9 mg, 0.021 mmol) in dry DMF (0.5 mL), HATU (7.9 mg, 0.021 mmol) and DIPEA (0.008 mL, 0.043 mmol) were added and the mixture was stirred at room temperature for 15 minutes. Then, 2-1m (trifluoroacetate) (18.0 mg, 0.017 mmol) was added to the above mixture and the mixture was stirred at room temperature for 10 minutes. The resulting solution was purified by preparative HPLC (method: column: XBridge Prep C18 OBD 5um 19 × 150 mm; mobile phase: A-water (0.1% TFA): B-acetonitrile; flow rate: 20 mL / min), and the fraction was lyophilized to obtain 3-1 (5.5 mg, yield 26.7%) as a yellow solid.

[0322] MS (ESI) m / z: 1205.6 [M+H] + .

[0323] Examples 2-5 [ka] 4-((S)-2-((S)-3-methyl-2-(4-(5-(methylsulfonyl)-1,2,4-thiadiazole-3-yl)benzamide)butanamide)-5-ureidopentanamide)benzyl((S)-1-(((S)-1-(((3R,4S,5S)-1-((S)-2-((1R,2R)-3-(((1S,2R)-1-hydroxy-1-phenylpropane-2-yl)amino)-1-methoxy-2-methyl-3-oxopropyl)pyrrolidine-1-yl)-3-methoxy-5-methyl-1-oxoheptan-4-yl)(methyl)amino)-3-methyl-1-oxobutan-2-yl)amino)-3-methyl-1-oxobutan-2-yl)(methyl)carbamate(2-5)

[0324] To a solution of 2-3d (10 mg, 0.035 mmol) in dry DMF (1 mL), HATU (15 mg, 0.040 mmol) and DIPEA (9.1 mg, 0.070 mmol) were added. The mixture was stirred at room temperature for 15 minutes. Then, 2-5a (TFA salt, commercially available) (36 mg, 0.032 mmol) was added to the mixture and stirred at room temperature for another 10 minutes. The resulting solution was purified by preparative HPLC (Method: Column XBridge Prep C18 OBD 5 μm 19 × 150 mm; Mobile phase: A - Water (0.1% TFA): B - Acetonitrile; Flow rate: 20 mL / min), and the fraction was lyophilized to obtain 2-5 (14 mg, yield 31.8%) as a white solid.

[0325] MS (ESI) m / z: 1389.8 [M+H] + .

[0326] Examples 2-6 [ka] Step 1: Benzyl(5S,8S)-1-(9H-fluoren-9-yl)-5-isopropyl-8,14,14-trimethyl-3,6,9-trioxo-2,12-dioxa-4,7,10-triazapentadecane-15-oate(2-6c)

[0327] A white suspension mixture of 2-6a (300 mg, 0.623 mmol), 2-6b (259.6 mg, 1.246 mmol), and 4 Šmolecular sieves in anhydrous THF (10 mL) was stirred at room temperature for 10 minutes. Sc(OTf)3 (368.0 mg, 0.748 mmol) was added, and the resulting yellow suspension was stirred at room temperature for 4 hours. The yellow suspension mixture was filtered through a Celite pad and washed with  (30 mL). The combined organic layers were washed with saturated NaHCO3 (30 mL) and brine (30 mL), dried over Na2SO4, filtered, and the filtrate was concentrated under vacuum to obtain the residue. This was purified by silica gel column (MeOH / DCM = 0%~5%), and the fraction was concentrated under vacuum to obtain 2-6c (274 mg, yield 69.8%) as a white solid.

[0328] MS (ESI) m / z: 652.6 [M+Na] + .

[0329] Step 2: Benzyl 3-(((S)-2-((S)-2-amino-3-methylbutanamide)propanamide)methoxy)-2,2-dimethylpropanoate (2-6d)

[0330] Et2N (477.3 mg, 5.53 mmol) was added to a solution of 2-6c (274.0 mg, 0.44 mmol) in DMF (5 mL). The mixture was stirred at room temperature for 20 minutes. The reaction mixture was concentrated under vacuum and evaporated twice with toluene to obtain 2-6d (275.3 mg, crude) as a brown oil.

[0331] MS (ESI) m / z: 430.4 [M+Na] + .

[0332] Step 3: Benzyl(5S,8S,11S)-5-(3-((((2R,3S,4R,5S)-5-(2-amino-2-oxoethyl)-3,4-dihydroxytetrahydrofuran-2-yl)methyl)amino)-3-oxopropyl)-1-(9H-fluoren-9-yl)-8-isopropyl-11,17,17-trimethyl-3,6,9,12-tetraoxo-2,15-dioxa-4,7,10,13-tetraazaoctadecane-18-oate(2-6f)

[0333] To a solution of 2-6d (275.3 mg, crude) and 2-6e (282.3 mg, 0.52 mmol) in DMF (5 mL), HATU (198.2 mg, 0.52 mmol) and DIPEA (168.4 mg, 1.30 mmol) were added. The mixture was stirred at room temperature for 10 minutes. The mixture was purified by reverse-phase chromatography (C18, 60 g, 30%-70%), and the fraction was freeze-dried to obtain 2-6f (370 mg, yield 91.5%) as a brown solid.

[0334] MS (ESI) m / z: 953.8 [M+Na] + .

[0335] Step 4: (5S,8S,11S,17R)-5-(3-((((2R,3S,4R,5S)-5-(2-amino-2-oxoethyl)-3,4-dihydroxytetrahydrofuran-2-yl)methyl)amino)-3-oxopropyl)-1-(9H-fluoren-9-yl)-17-fluoro-8-isopropyl-11,17-dimethyl-3,6,9,12-tetraoxo-2,15-dioxa-4,7,10,13-tetraazaoctadecane-18-euic acid (2-6g)

[0336] Compound 2-6f (3.0 g, 3.21 mmol) was dissolved in cosolvent DMF-MeOH (40 mL, 1:1, v:v) and Pd / C (10%, 600 mg) was added. The mixture was stirred under an H2 atmosphere (15 psi) for 7 hours. The mixture was filtered through a Celite pad and concentrated to obtain compound 2-6 g (2.5 g, crude) as a white solid.

[0337] MS (ESI) m / z: 863.7 [M+Na] + .

[0338] Step 5: (9H-Fluoren-9-yl)methyl((6S,9S,12S)-1-((2R,3S,4R,5S)-5-(2-amino-2-oxoethyl)-3,4-dihydroxytetrahydrofuran-2-yl)-19-(((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10 ,13,15-Hexahydro-1H,12H-Benzo[de]pyrano[3',4':6,7]indolidino[1,2-b]quinoline-1-yl)amino)-9-isopropyl-12,18,18-trimethyl-3,7,10,13,19-pentaoxo-16-oxa-2,8,11,14-tetraazanonadecane-6-yl)carbamate(2-6h)

[0339] To a solution of exatecan mesylate (1000 mg, 1.18 mmol) in DMF (20 mL), 2-6 g of the compound (692 mg, 1.3 mmol), HATU (675 mg, 1.78 mmol), and DIPEA (459 mg, 3.55 mmol) were added. The mixture was stirred at room temperature for 30 minutes. The mixture was concentrated and purified by silica gel column chromatography (eluent: DCM / MeOH = 100 / 0~20 / 80) to obtain the title compound 2-6 h (1320 mg, yield 88.6%) as an off-white solid.

[0340] MS (ESI) m / z: 1282.1 [M+Na] + .

[0341] Step 6: (S)-2-amino-N5-(((2R,3S,4R,5S)-5-(2-amino-2-oxoethyl)-3,4-dihydroxytetrahydrofuran-2-yl)methyl)-N1-((S)-1-(((S)-1-(((3-(((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3 ,9,10,13,15-Hexahydro-1H,12H-benzo[de]pyrano[3',4':6,7]indolidino[1,2-b]quinoline-1-yl)amino)-2,2-dimethyl-3-oxopropoxy)methyl)amino)-1-oxopropan-2-yl)amino)-3-methyl-1-oxobutan-2-yl)pentanediamide(2-6i)

[0342] Et2NH (580 mg, 7.93 mmol) was added to a solution of compound 2-6h (1000 mg, 0.793 mmol) in DMF (20 mL). The mixture was stirred at room temperature for 30 minutes. The mixture was concentrated under high vacuum to obtain compound 2-6i (824.6 mg, crude) as an off-white solid, which was used directly without further purification.

[0343] MS (ESI) m / z: 1036.9 [M+H] + .

[0344] Step 7: (S)-N5-(((2R,3S,4R,5S)-5-(2-amino-2-oxoethyl)-3,4-dihydroxytetrahydrofuran-2-yl)methyl)-N1-((S)-1-(((S)-1-(((3-(((1S,9S)-9-ethyl-5-fluoro-9-hydroxy-4-methyl-10,13-dioxo-2,3,9,10,13,15-hexahydro-1H,12H -Benzo[de]pyrano[3',4':6,7]indolidino[1,2-b]quinoline-1-yl)amino)-2,2-dimethyl-3-oxopropoxy)methyl)amino)-1-oxopropan-2-yl)amino)-3-methyl-1-oxobutan-2-yl)-2-(4-(5-(methylsulfonyl)-1,2,4-thiadiazole-3-yl)benzamide)pentanediamide(2-6)

[0345] To a solution of 2-3d (15.2 mg, 0.053 mmol) in dry DMF (1.0 mL), HATU (22.2 mg, 0.058 mmol) and DIPEA (0.017 mL, 0.097 mmol) were added and the mixture was stirred at room temperature for 15 minutes. Then, 2-6i (50.0 mg, 0.048 mmol) was added to the above mixture and the mixture was stirred at room temperature for 10 minutes. The resulting solution was purified by preparative HPLC (method: column: XBridge Prep C18 OBD 5um 19 × 150 mm; mobile phase: A-water (0.1% TFA): B-acetonitrile; flow rate: 20 mL / min), and the fraction was lyophilized to obtain 2-6 (32 mg, yield 50.9%) as a yellow solid.

[0346] MS (ESI) m / z: 1303.0 [M+H] + .

[0347] The compounds from Examples 1-1 to 1-5 are shown in Table 1 below. The compounds from Examples 2-1 to 2-6 are shown in Table 2 below. Preparation of conjugator-antibody conjugates and antibody-drug conjugates

[0348] Preparation of DAR8 antibody-drug conjugates / conjugator-antibody conjugates. Antibody (concentration 0.5–25 mg / mL, PBS buffer pH 6.0–8.5) in conjugation buffer was incubated at reducing temperature (0–40°C) for 10 minutes. 8–15 equivalents of TECP solution (5 mM stock in PBS buffer) were added to the reaction mixture, and the reduction reaction was allowed to stand at reducing temperature for 1–8 hours. After cooling the reducing mixture to 0–25°C, organic solvents (e.g., DMSO, DMF, DMA, PG, acetonitrile, 0–25% v / v) and conjugator-linker-payload (see Table 2) or conjugator (see Table 1) stock (10–25 equivalents, 10 mM stock in organic solvent) were added stepwise. The conjugation solution was allowed to stand at 0–25°C for 1–3 hours, and the reaction was quenched with N-acetylcysteine ​​(1 mM stock). The buffer solution was replaced with a storage buffer (for example, histidine acetate buffer with a pH of 5.5-6.5 to which optional additives (e.g., sucrose, trehalose, tween® 20, 60, 80) was added) (by spin desalination column, ultrafiltration, and dialysis).

[0349] Tables 3 and 4 show the conjugator-antibody conjugates and ADCs prepared according to the above method.

[0350] Characterization of ADCs. The following analytical methods were used to characterize the ADCs. The drug-to-antibody ratio (DAR) of the ADCs was quantified by LC-MS or HIC. The SEC purity of all prepared ADCs was over 95%.

[0351] LC-MS method: LC-MS analysis was performed under the following measurement conditions. LC-MS system: Vanquish Flex UHPLC and Orbitrap Exploris 240 mass spectrometer Column: MAbPac® RP, 2.1 × 50 mm, 4 μm, 1,500 Å, Thermo Scientific® Column temperature: 80℃ Mobile phase A: 0.1% formic acid (FA) aqueous solution Mobile phase B: Acetonitrile solution containing 0.1% formic acid (FA) Gradient Programs: 25% B~25% B (0 min~2 min), 25% B~50% B (2 min~18 min), 50% B~90% B (18 min~18.1 min), 90% B~90% B (18.1 min~20 min), 90% B~25% B (20 min~20.1 min), 25% B~25% B (20.1 min~25 min) Injection sample volume: 1 μg MS parameters: Intact and denatured MS data were acquired in HMR mode with R=15k and deconvolved using the ReSpect algorithm and sliding window integration with Thermo Scientific® BioPharma Finder® 4.0 software.

[0352] HIC method: HPLC analysis was performed under the following measurement conditions. Method 1 HPLC System: Waters ACQUITY ARC HPLC System Detector: Measurement wavelength: 280nm Column: Tosoh Bioscience 4.6μm ID×3.5cm, 2.5μm butyl nonporous resin column Column temperature: 25℃ Mobile phase A: 1.5M ammonium sulfate, 50mM phosphate buffer, pH 7.0 Mobile phase B: 50 mM phosphate buffer, 25% (V / V) isopropanol, pH 7.0 Gradient Programs: 0% B~0% B (0 min~2 min), 0% B~100% B (2 min~15 min), 100% B~100% B (15 min~16 min), 100% B~0% B (16 min~17 min), 0% B~0% B (17 min~20 min) Injection sample volume: 20 μg Method 2 HPLC System: Waters ACQUITY ARC HPLC System Detector: Measurement wavelength: 280nm Column: MABPac HIC-10, 5μm, 4.6×10mm (Thermo) Column temperature: 25℃ Mobile phase A: 1.5M ammonium sulfate, 50mM sodium phosphate, pH 7.0 Mobile phase B: 50 mM sodium phosphate, pH 7.0 Gradient program: 20% B to 20% B (0 min to 1 min), 0% B to 0% B (1 min to 35 min), 20% B to 20% B (35 min to 40 min) Flow rate: 0.5mL / min Sample preparation: The sample was diluted with the initial mobile phase to a concentration of 0.5 mg / mL.

[0353] SEC method: HPLC analysis was performed to quantify ADC purity under the following measurement conditions. HPLC System: Waters H-Class UPLC System Detector: Measurement wavelength: 280nm Column: ACQUITY UPLC BEH200 SEC 1.7um 4.6×150mm, Waters Column temperature: Room temperature Mobile phase A: 200 mM phosphate buffer, 250 mM potassium chloride, 15% isopropyl alcohol, pH 7.0 Gradient program: 10 minutes of isocratic elution, flow rate 0.3 mL / min Injection sample volume: 20 μg

[0354] Hydrophobicity Evaluation of ADCs: The hydrophobicity of ADCs was evaluated using the HIC (hydrophobic interaction column) chromatography method described above. It is expected that ADCs with high hydrophobicity will show up in the HIC chromatography with a slow retention time. The results using the DAR8 peak as the reference are shown in Table 4.

[0355] [Table 1]

[0356] [Table 2-1] [Table 2-2]

[0357] [Table 3]

[0358] [Table 4-1] [Table 4-2]

[0359] Antibody information Ifinatamab (anti-B7H3 antibody)

[0360] Light chain sequence (SEQ ID NO: 1) EIVLTQSPATLSLSPGERATLSCRASSRLIYMHWYQQKPGQAPRPLIYATSNLASGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCQQWNSNPPTFGQGTKVEIK RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC

[0361] Heavy chain sequence (SEQ ID NO: 2) QVQLVQSGAEVKKPGSSVKVSCKASGYTFTNYVMHWVRQAPGQGLEWMGYINPYNDDVKYNEKFKGRVTITADESTSTAYMELSSLRSEDTAVYYCARWGYYGSPLYYFDYWG QGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCD KTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEK TISKAKGQPREPQVYTLPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK

[0362] 6E7 (anti-CLL antibody)

[0363] Light chain sequence (SEQ ID NO: 3) DIQMTQSPSSLSASVGDRVTITCRASQSVSTSSYNYMHWYQQKPGKPPKLLIKYASNLESGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQHSWEIPLTFGQGTKVE IKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC

[0364] Heavy chain sequence (SEQ ID NO: 4) EVQLVQSGAEVKKPGASVKVSCKASGYSFTDYYMHWVRQAPGQGLEWIGRINPYAGAAFYSQNFKDRVTLTVDTSTSTAYLELSSLRSEDTAVYYCAIERGADLEGYAMDYWG QGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCD KTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEK TISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK Conjugator-antibody conjugate stability test: ADC storage buffer and GSH solution

[0365] In these studies, the stability of conjugator-antibody conjugates in typical ADC acidic storage buffers was investigated, as well as the deconjugation event when the conjugates were incubated in glutathione (GSH) solution. Specifically, after the conjugation step described above (see "Preparation of DAR8 Antibody-Drug Conjugates / Conjugator-Antibody Conjugates"), the conjugator-antibody conjugates were incubated in formulation buffer (pH 5.5 20 mM histidine buffer) or GSH buffer (pH 7.4 or 8.0), and the solutions were left at 22 or 37°C for 1 to 168 hours. The results are shown in Tables 5 and 6, and Figures 1 to 10. [Table 19]

[0366] [Table 5]

[0367] The results show that the DAR values ​​of all conjugates remained nearly the same as T0 even after incubation for at least 18 hours in GSH buffer (pH 7.4, 22 / 37°C and pH 8.0, 22°C).

[0368] [Table 6]

[0369] The results indicate that the DAR values ​​of all conjugates remained nearly the same as T0 even after incubation in formulation buffer for 168 hours.

[0370] The results of the stability evaluation of ADCs in GSH solution or formulation buffer are shown in Tables 7 and 8, and Figures 11-16, respectively.

[0371] [Table 7]

[0372] The results shown in Table 7 indicate that no deconjugation events were observed in ADC 4-1 to 4-3 after incubation with GSH buffer (pH 7.4, 22 / 37°C and pH 8.0, 22°C).

[0373] [Table 8]

[0374] The results shown in Table 8 indicate that no deconjugation events were observed in ADCs 4-1 to 4-3 after storage in formulation buffer for one week.

[0375] cell line NCI-H1650 (ATCC, CRL-5883). NCI-H1650 is an epithelial cell line isolated in 1987 from the lung tissue of a 27-year-old male smoker with stage 3 bronchoalveolar carcinoma. NCI-H1650 was purchased from ATCC. The basic medium for NCI-H1650 was ATCC-formulated RPMI-1640 medium (ATCC 30-2001). To prepare a full-growth medium, 10% final concentration fetal bovine serum (Gibco, 10099-141C) was added to the basic medium. The cell line was grown at 37°C in a humidified 5% CO2 atmosphere, and the presence or absence of mycoplasma was checked periodically using the MycoAlert® PLUS Mycoplasma Detection Kit (Lonza, LT07-710).

[0376] Capan-1 (ATCC, HTB-79). Capan-1 is an epithelial cell line isolated from the pancreas of a 40-year-old Caucasian male with pancreatic adenocarcinoma, and was purchased from ATCC. The basic medium for Capan-1 was ATCC-formulated Iskov modified Dulbecco's medium (catalog no. 30-2005). To prepare a full-growth medium, fetal bovine serum (Gibco, 10099-141C) at a final concentration of 20% was added to the basic medium. The cell line was grown at 37°C in a humidified 5% CO2 atmosphere, and the presence or absence of mycoplasma was checked periodically using the MycoAlert® PLUS Mycoplasma Detection Kit (Lonza, LT07-710).

[0377] MDA-MB-453 (SIBS). MDA-MB-453 was derived from the pleural effusion of a 48-year-old female patient with metastatic breast cancer that had invaded the lymph nodes, brain, and both the pleural and pericardial cavities. MDA-MB-453 was purchased from SIBS. The basic medium for MDA-MB-453 was RPMI 1640 medium HEPES (Gibco; 22400105). To prepare a full-growth medium, 10% final concentration fetal bovine serum (Gibco, 10099-141C) was added to the basic medium. The cell line was grown at 37°C in a humidified 5% CO2 atmosphere, and the presence or absence of mycoplasma was checked periodically using the MycoAlert® PLUS Mycoplasma Detection Kit (Lonza, LT07-710).

[0378] U937 (ATCC, CRL-1593.2). U-937 is a cell line exhibiting monocyte morphology, derived from malignant cells obtained from the pleural effusion of a 37-year-old Caucasian male with histiocytic lymphoma in 1974. U937 was purchased from ATCC. The basic medium for U937 was ATCC-formulated RPMI-1640 medium (ATCC 30-2001). To prepare a full-growth medium, fetal bovine serum (Gibco, 10099-141C) at a final concentration of 10% was added to the basic medium. The cell line was grown at 37°C in a humidified 5% CO2 atmosphere, and the presence or absence of mycoplasma was checked periodically using the MycoAlert® PLUS Mycoplasma Detection Kit (Lonza, LT07-710).

[0379] HL60 (ATCC, CCL-240). HL-60 cells are promyeloblasts isolated by leukophagesis from the peripheral blood of a 36-year-old Caucasian woman with acute promyelocytic leukemia. HL60 cells were purchased from ATCC. The basic medium for HL60 was ATCC-formulated Iskov modified Dulbecco's medium (catalog number 30-2005). To prepare a complete growth medium, 20% final concentration fetal bovine serum (Gibco, 10099-141C) was added to the basic medium. The cell line was grown at 37°C in a humidified 5% CO2 atmosphere, and the presence or absence of mycoplasma was checked periodically using the MycoAlert® PLUS Mycoplasma Detection Kit (Lonza, LT07-710).

[0380] TF1 (ATCC, CRL-2003). TF-1 erythroblasts were isolated in 1987 from the bone marrow of a 35-year-old Asian male with severe pancytopenia. TF-1 was purchased from ATCC. The basic medium for TF-1 was ATCC-formulated RPMI-1640 medium (catalog number 30-2001). To prepare a full-growth medium, 10% final concentration fetal bovine serum (Gibco, 10099-141C) was added to the basic medium. The cell line was grown at 37°C in a humidified 5% CO2 atmosphere, and the presence or absence of mycoplasma was checked periodically using the MycoAlert® PLUS Mycoplasma Detection Kit (Lonza, LT07-710).

[0381] NCI-H358 (ATCC, CRL-5807). NCI-H358 cells are epithelial-like cells isolated from the bronchi of a male patient with bronchoalveolar carcinoma. NCI-H358 was purchased from ATCC. The basic medium for NCI-H358 was ATCC-formulated RPMI-1640 medium (ATCC 30-2001). To prepare a full-growth medium, 10% final concentration fetal bovine serum (Gibco, 10099-141C) was added to the basic medium. The cell line was grown at 37°C in a humidified 5% CO2 atmosphere, and the presence or absence of mycoplasma was checked periodically using the MycoAlert® PLUS Mycoplasma Detection Kit (Lonza, LT07-710). [Table 10A] [Table 10B]

[0382] Direct killing of ADCs in U937, HL60, TF1, NCI-H1650, Capan-1, and MDA-MB-453 cancer cell lines Direct ADC killing was evaluated in U937, HL60, and TF1 cancer strains. Cells (U937 or HL60 (3E3 / well) or TF1 (6E3 / well)) were seeded in 100 μl / well (containing 150 μg / ml of Fc blocker) in 2D 96-well plates (Greiner: 655090) and incubated overnight at 37°C, 5% CO2. 50 μl / well of fresh growth medium containing various concentrations of ADC was added and incubated at 37°C, 5% CO2 for 6 days. Cell viability was detected using Cell Titer-Glo (Promega, G7573), 70 μl / well. The 2D plates were incubated at room temperature for 10 minutes to stabilize the luminescence signal. The plates were analyzed using a microplate reader.

[0383] Direct ADC killing was evaluated in NCI-1650, Capan-1, and MDA-MB-453 cancer strains. Cells (NCI-1650 or MDA-MB-453 (2E3 / well) or Capan-1 (4E3 / well)) were seeded at 80 μl / well in 3D 96-well plates (Corning: 4520) and incubated overnight at 37°C, 5% CO2. 40 μl / well of fresh growth medium containing various concentrations of ADC was added, and the plates were incubated at 37°C, 5% CO2 for 6 days. Cell viability was detected using 3D reagent (Promega; G9683), 100 μl / well. The 3D plates were incubated at room temperature for 30 minutes to stabilize the luminescence signal. The plates were analyzed using a microplate reader.

[0384] The data is summarized in Figures 17-25 and Tables 11-13. [Table 11] [Table 12] [Table 13]

[0385] Bystander killing of ADCs in co-culture of NCI-H358 and MDA-MB-453-nanoLuc Methods: Construction of the MDA-MB-453-nanoLuc cell line. PT67-nanoLuc cells were cultured, then the cell culture medium (containing the virus (nano-Luc gene)) was collected and filtered. MDA-MB-453 cells were seeded at 1E5 cells / well in a 6-well plate and incubated overnight at 37°C and 5% CO2. PT67-nanoLuc cell medium and 8 μg / ml of polyblen were added. Infection was repeated three times every other day. Then, 1 mg / ml of geneticin was added to the MDA-MB-453-nanoLuc cells and they were cultured for 5 days. The MDA-MB-453-nanoLuc cells were collected, and the nano-Luc transfection efficiency was tested by adding Nano-Glo reagent (Promega: N1120).

[0386] Methods: Bystander killing of ADCs. NCI-H358 and MDA-MB-453-nanoLuc (10:1), or MDA-MB-453-nanoLuc cells alone, were seeded at 80 μl / well in 3D-96 well plates (Corning: 4520) and incubated overnight at 37°C, 5% CO2. Fresh growth medium containing various concentrations of ADC was added at 40 μl / well. Cells were incubated at 37°C, 5% CO2 for 6 days. The 3D plates were centrifuged at 1500 rpm, 25°C, for 5 minutes, and the supernatant was discarded.

[0387] Calu-6-nanoLuc cell viability was detected using Nano-Glo reagent (Promega:N1120), 150 ul / well. 3D plates were incubated at room temperature for 10 minutes to stabilize the luminescence signal. The plates were then analyzed using a microplate reader.

[0388] The results are shown in Tables 14-15 and Figures 26-29. [Table 14] [Table 15]

[0389] In vivo efficacy study of ADC in the H1650 xenograft model In the right ventral region of female BALB / c nude mice, 3 × 10¹⁴ cells were placed per 200 μL of PBS / Matrigel. 6 H1650 cells were subcutaneously transplanted. After inoculation, tumor volume was quantitatively measured two-dimensionally twice a week using calipers, and the formula was: V = 0.5(a × b 2 ) using mm 3 This was represented as follows: The tumor size was approximately 200 mm. 3Once the average volume was reached, the mice were randomly assigned to three groups of seven mice each and administered intravenously with either the vehicle or ADC 4-B, 4-2, 4-3, or 4-4 at a dose of 1 mpk or 3 mpk twice weekly. Partial regression (PR) was defined as three consecutive tumor volume measurements being less than 50% of the initial tumor volume on the first day of administration, and complete regression (CR) was defined as three consecutive tumor volume measurements of 14 mm. 3 It was defined as being smaller than the mean. The data is shown as mean tumor volume ± standard error of the mean (SEM). Tumor growth inhibition rate (TGI) is calculated using the following formula.

number

[0390] The results are shown in Figures 30 and 31. The results show that ADC 4-2, 4-3, and 4-4 each exhibited good antitumor activity (Figures 30 and 31). ADC 4-2 showed better antitumor activity than the reference ADC 4-B (Figure 30). ADC 4-3 and 4-4 showed comparable antitumor activity (Figure 31). ADC 4-3 showed dose-dependent efficacy (Figure 31).

[0391] Plasma stability of ADCs Incubation of ADC with plasma: ADC was diluted in mouse or human plasma to obtain a final solution of 100 μg / mL ADC in plasma. Samples were incubated at 37°C. Aliquots (100 μL) were taken at five time points (0, 2(4), 24, 72, or 168 hours). Samples were frozen at -80°C until analysis.

[0392] Plasma payload concentrations were measured under the following conditions. Equipment: LC-MS / MS (Triple Quad 6500 plus) Monitor: MRM Column: Advanced Materials Technology, HALO AQ-C18 2.7μm 90Å, 50×2.1mm Column temperature: 40℃ Mobile phase A: H2O - 0.1% FA Mobile phase B: ACN-0.1% FA MMAE gradient programs: 15% B~15% B (0 min~0.4 min), 15% B~30% B (0.4 min~0.8 min), 30% B~30% B (0.8 min~1.8 min), 30% B~90% B (1.8 min~1.9 min), 90% B~90% B (1.9 min~2.4 min), 90% B~15% B (2.4 min~2.5 min), 15% B~15% B (2.5 min~3.0 min) DXd gradient programs: 2% B~2% B (0 min~0.2 min), 2% B~98% B (0.2 min~1.2 min), 98% B~98% B (1.2 min~2.0 min), 98% B~2% B (2.0 min~2.01 min), 2% B~2% B (2.01 min~4.0 min) Injection sample volume: 10 μL (DXd or DXd analog or MMAE)

[0393] The results are shown in Figures 32-35. ADCs 4-1 to 4-4 showed lower payload release percentages than ADC-4-B in both mouse plasma (Figure 32) and human plasma (Figure 33). ADC 4-5 showed a payload release rate comparable to ADC 4-A in mouse plasma (Figure 34), and a higher payload release rate than ADC 4-A in human plasma (Figure 35).

[0394] Mouse PK Test 1 After a single intravenous administration of ADC (3 mg / kg) to H1650 tumor-bearing mice, blood samples were collected at 0.0833, 2, 24, 72, 120, and 168 hours, followed by centrifugation (4°C, 3000xg, 7 min) to separate the plasma. The ADC concentration was measured using the in-house developed Meso Scale Discovery (MSD) ligand-binding method. Briefly, a His-tagged B7H3 extracellular domain fusion protein was used as the capture reagent, and biotin-labeled anti-payload Ab was used as the ADC detection reagent. The plasma concentration of the payload was measured using the same method as described above.

[0395] The results are shown in Figure 36. The results indicate that ADC 4-2 to 4-4 showed a PK profile equivalent to that of ADC 4-B in the H1650 efficacy model.

[0396] Mouse PK Test 2 Non-tumor-bearing mice were given a single intravenous dose of ADC (5 mg / kg), and blood samples were collected at 0.5, 2, 8, 24, 72, and 168 hours. Plasma was then separated by centrifugation (4°C, 3000xg, 7 min). The concentration of ADC was measured using the in-house developed Meso Scale Discovery (MSD) ligand binding method. Briefly, a His-tagged CLL1 extracellular domain fusion protein was used as the capture reagent, and biotin-labeled anti-payload Ab was used as the detection reagent for ADC. The plasma concentration of the payload was measured using the same method as described above.

[0397] The results are shown in Figure 37. The results indicate that ADC 4-5 exhibited a PK profile equivalent to that of ADC 4-A in non-tumor-bearing mice.

[0398] Although the above disclosures are presented in some detail through descriptions and examples for the purpose of clarification, it will be apparent to those skilled in the art that certain minor changes and modifications will be made. Therefore, this specification and the examples should not be construed as limiting.

[0399] Where any publication is referenced herein, please understand that such reference does not constitute an admission that such publication constitutes part of the general knowledge in the art in any country.

[0400] All non-patent publications, patents, patent applications, and published patent applications referred to herein are incorporated herein by reference in their entirety.

Claims

1. Compound of formula (I): 【Chemistry 51】 or a pharmaceutically acceptable salt, tautomer, solvate, or stereoisomer thereof. [In the formula, BA is a binder selected from humanized antibodies, chimeric antibodies, human antibodies, or their antigen-binding fragments. U is an arylene, heteroarylene, or bond. V is a bond or -C≡C-(CH ２ ) ｎ - and n is an integer between 0 and 10. A is a residue in the stretcher unit, The subscript a' is either 0 or 1. W is a cleaveable unit, The subscript 'w' is either 0 or 1. Y is a spacer unit, The subscript y' is either 0 or 1. PA is payload residue, The subscript x is between 1 and 15.

2. The compound according to claim 1, wherein U is arylene.

3. The compound according to claim 2, wherein U is phenylene.

4. U 【Chemistry 52】 The compound according to claim 3.

5. The compound according to any one of claims 1 to 4, wherein V is a bond.

6. V is -C ≡ C - (CH ２ ) ｎ - The compound according to any one of claims 1 to 4.

7. V is -C ≡ C - (CH ２ ) ３ - The compound according to claim 6.

8. The compound according to claim 1, wherein U is a heteroarylene.

9. The compound according to claim 8, wherein U is a divalent pyrimidine ring.

10. U 【Chemistry 53】 The compound according to claim 9.

11. The compound according to any one of claims 8 to 10, wherein V is a bond.

12. V is -C ≡ C - (CH ２ ) ｎ - The compound according to any one of claims 8 to 10.

13. V is -C≡C-(CH ２ ), ３ The compound according to claim 12, wherein

14. The compound according to claim 1, wherein U is a bond.

15. V is -C ≡ C - (CH ２ ) ｎ - The compound according to claim 14.

16. V is -C ≡ C - (CH ２ ) ３ - The compound according to claim 15.

17. Aが、-(CH ２ ) ｍ -C(=O)-、-CH ２ -C(=O)-NH-(CH ２ ) ｍ -C(=O)-、 -(CH ２ CH ２ O) ｍ -CH ２ CH ２ -C(=O)-、-CH[-(CH ２ ) ｍ -COOH]-C(=O)-、 -CH ２ -C(=O)-NH-(CH ２ ) ｍ -C(=O)-NH-(CH ２ ) ｍ -C(=O)-、 -C(=O)-(CH ２ ) ｍ -C(=O)-, -NH-(CH ２ ) ｍ -C(=O)-, or -NH-(CH ２ CH ２ O) ｍ -CH ２ CH ２ The compound according to any one of claims 1 to 16, wherein -C(=O)-, and each m independently represents an integer of 1, 2, 3, 4, or 5.

18. The compound according to any one of claims 1 to 17, wherein the subscript a' is 0.

19. The compound according to any one of claims 1 to 17, wherein the subscript a' is 1.

20. W is one of the following equations: 【Chemistry 54】 The compound according to any one of claims 1 to 19, wherein HG is a hydrophilic moiety or hydrogen.

21. The compound according to claim 20, wherein HG is a saccharide, a phosphate ester, a sulfate ester, a phosphodiester, or a phosphonate.

22. The compound according to claim 21, wherein HG is a saccharide, and the saccharide is β-D-galactose, N-acetyl-P-D-galactosamine, N-acetyl-a-D-galactosamine, N-acetyl-P-D-glucosamine, β-D-glucuronic acid, a-L-iduronic acid, a-D-galactose, a-D-glucose, β-D-glucose, a-D-mannose, β-D-mannose, a-L-fucose, β-D-xylose, neuraminic acid, or sulfate, phosphate, carboxyl, amino, or O-acetyl modified products thereof.

23. HG, 【Transformation 55】 The compound according to claim 20.

24. The compound according to any one of claims 1 to 23, wherein the subscript w' is 1.

25. The compound according to any one of claims 1 to 23, wherein the subscript w' is 0.

26. Y is -NH-CH ２ -O- or -NH- (p-C) ６ H ４ ) - CH ２ A compound according to any one of claims 1 to 25, wherein the compound is -O-.

27. The compound according to any one of claims 1 to 26, wherein the subscript y' is 1.

28. The compound according to any one of claims 1 to 26, wherein the subscript y' is 0.

29. The compound according to any one of claims 1 to 28, wherein the compound is one of the following formulas. 【Transformation 56】

30. portion 【Chemistry 57】 However, the compound according to any one of claims 1 to 16 and 29 is one of the following formulas. 【Chemistry 58-1】 【Chemistry 58-2】

31. The compound according to any one of claims 1 to 30, wherein BA is ifinatamab, 6E7, or trastuzumab, or an antigen-binding fragment of ifinatamab, 6E7, or trastuzumab.

32. The compound according to any one of claims 1 to 30, wherein BA is a humanized, chimeric, or human antibody, or an antigen-binding fragment thereof, that binds to one or more receptors selected from HER2, CLL1, or B7H3.

33. The aforementioned compound, 【Chemistry 59】 The compound according to claim 1, or a pharmaceutically acceptable salt, tautomer, or solvate thereof, wherein Ab is ifinatamab.

34. The aforementioned compound, 【Transformation 60】 The compound according to claim 1, or a pharmaceutically acceptable salt, tautomer, or solvate thereof, wherein Ab is 6E7.

35. A compound according to any one of claims 1 to 34, or a pharmaceutically acceptable salt thereof, tautomorphism A pharmaceutical composition comprising a compound, a solvate, or a stereoisomer, and a pharmaceutically acceptable excipient.

36. Compound of formula (II): 【Chemistry 61】 or a pharmaceutically acceptable salt, tautomer, solvate, or stereoisomer thereof. [In the formula, U is an arylene, heteroarylene, or bond. V is a bond or -C≡C-(CH ２ ) ｎ - and n is an integer between 0 and 10. A is a residue in the stretcher unit, The subscript a' is either 0 or 1. W is a cleaveable unit, The subscript 'w' is either 0 or 1. Y is a spacer unit, The subscript y' is either 0 or 1. PA is a payload residue.

37. The compound according to claim 36, wherein U is arylene.

38. The compound according to claim 37, wherein U is phenylene.

39. U 【Transformation 62】 The compound according to claim 38.

40. The compound according to any one of claims 36 to 39, wherein V is a bond.

41. V is -C ≡ C - (CH ２ ) ｎ - The compound according to any one of claims 36 to 39.

42. V is -C ≡ C - (CH ２ ) ３ - The compound according to claim 41.

43. The compound according to claim 36, wherein U is a heteroarylene.

44. The compound according to claim 43, wherein U is a divalent pyrimidine ring.

45. U 【Transformation 63】 The compound according to claim 44.

46. The compound according to any one of claims 43 to 45, wherein V is a bond.

47. V is -C ≡ C - (CH ２ ) ｎ - The compound according to any one of claims 43 to 45.

48. V is -C ≡ C - (CH ２ ) ３ - The compound according to claim 47.

49. The compound according to claim 36, wherein U is a bond.

50. V is -C ≡ C - (CH ２ ) ｎ - The compound according to claim 49.

51. V is -C ≡ C - (CH ２ ) ３ - The compound according to claim 50.

52. A, -(CH) ２ ) ｍ -C(=O)-、-CH ２ -C(=O)-NH-(CH ２ ) ｍ -C(=O)-、 -(CH ２ CH ２ O) ｍ -CH ２ CH ２ -C(=O)-、-CH[-(CH ２ ) ｍ -COOH]-C(=O)-、 -CH ２ -C(=O)-NH-(CH ２ ) ｍ -C(=O)-NH-(CH ２ ) ｍ -C(=O)-、-C(=O)-(CH ２ ) ｍ -C(=O)-、 -NH-(CH ２ ) ｍ -C(=O)-, or -NH-(CH ２ CH ２ O) ｍ -CH ２ CH ２ The compound according to any one of claims 36 to 51, wherein -C(=O)-, and each m independently represents an integer of 1, 2, 3, 4, or 5.

53. The compound according to any one of claims 36 to 52, wherein the subscript a' is 0.

54. The compound according to any one of claims 36 to 52, wherein the subscript a' is 1.

55. W is one of the following equations: 【Chemistry 64】 The compound according to any one of claims 36 to 54, wherein HG is a hydrophilic moiety or hydrogen.

56. The compound according to claim 55, wherein HG is a saccharide, a phosphate ester, a sulfate ester, a phosphodiester, or a phosphonate.

57. HG is a saccharide, and the saccharide is β-D-galactose, N-acetyl-P-D-galactosamine, N-acetyl-a-D-galactosamine, N-acetyl-P-D-glucosamine, β-D-glucuronic acid, a-L-iduronic acid, a-D-galactose, a-D-gluco The compound according to claim 56, which is β-D-glucose, α-D-mannose, β-D-mannose, α-L-fucose, β-D-xylose, neuraminic acid, or a sulfate, phosphate, carboxyl, amino, or O-acetyl modified product thereof.

58. HG, 【Transformation 65】 The compound according to claim 55.

59. The compound according to any one of claims 36 to 58, wherein the subscript w' is 1.

60. The compound according to any one of claims 36 to 58, wherein the subscript w' is 0.

61. Y is -NH-CH ２ -O- or -NH- (p-C) ６ H ４ ) - CH ２ A compound according to any one of claims 36 to 60, wherein it is -O-.

62. The compound according to any one of claims 36 to 61, wherein the subscript y' is 1.

63. The compound according to any one of claims 36 to 61, wherein the subscript y' is 0.

64. The compound according to any one of claims 36 to 63, wherein the compound is one of the following formulas. 【Chemical Formula 66】

65. portion 【Transformation 67】 However, the compound according to any one of claims 36 to 51 and 64 is one of the following formulas. 【Chemistry 68-1】 【Chemistry 68-2】

66. The aforementioned compound, 【Transformation 69】 The compound according to claim 65, or a pharmaceutically acceptable salt, tautomer, or solvate thereof.

67. The compound according to any one of claims 1 to 32 or 36 to 65, wherein each PA is independently a cytotoxic agent.

68. The compound according to claim 67, wherein each PA is independently selected from residues in the group consisting of DXd, 7-ethyl-10-hydroxycamptothecin (SN-38), and monomethyl auristatin E (MMAE).

69. Each PA is independently a compound of formula (VI), 【Transformation 70】 wherein R ９ and R １０ each independently is hydrogen, halogen, or substituted or unsubstituted C １～４ alkyl, the compound according to claim 68.

70. Each PA system operates independently. 【Chemistry 71】 The compound according to claim 69.

71. Compound of formula (III): 【Chemistry 72】 or a pharmaceutically acceptable salt, tautomer, solvate, or stereoisomer thereof. [In the formula, U is an arylene, heteroarylene, or bond. V is a bond or -C≡C-(CH ２ ) ｎ - and n is an integer between 0 and 10. A is a stretcher unit, The subscript a' is either 0 or 1.

72. The compound according to claim 71, wherein U is arylene.

73. The compound according to claim 72, wherein U is phenylene.

74. U 【Transformation 73】 The compound according to claim 73.

75. The compound according to any one of claims 71 to 74, wherein V is a bond.

76. V is -C ≡ C - (CH ２ ) ｎ - The compound according to any one of claims 71 to 74.

77. V is -C≡C-(CH ２ ), ３ a compound according to claim 76, wherein V is -C≡C-

78. The compound according to claim 71, wherein U is a heteroarylene.

79. The compound according to claim 78, wherein U is a divalent pyrimidine ring.

80. U 【Chemistry 74】 The compound according to claim 79.

81. The compound according to any one of claims 78 to 80, wherein V is a bond.

82. V is -C ≡ C - (CH ２ ) ｎ - The compound according to any one of claims 78 to 80.

83. V is -C ≡ C - (CH ２ ) ３ - The compound according to claim 82.

84. The compound according to claim 71, wherein U is a bond.

85. V is -C ≡ C - (CH ２ ) ｎ - The compound according to claim 84.

86. V is -C ≡ C - (CH ２ ) ３ - The compound according to claim 85.

87. A is a bond, -OH, -CH ３ , -N(CH ３ ) ２ ,-(CH ２ ) ｍ -C(=O)R ７ , -CH ２ -C(=O)-NH-(CH ２ ) ｍ -C(=O)R ７ 、 -(CH ２ CH ２ O) ｍ -CH ２ CH ２ -C(=O)R ７ 、 -CH[-(CH ２ ) ｍ -COOH]-C(=O)R ７ 、 -CH ２ -C(=O)-NH-(CH ２ ) ｍ -C(=O)-NH-(CH ２ ) ｍ -C(=O)R ７ 、-C(=O)-(CH ２ ) ｍ -C(=O)R ７ 、 -NH-(CH ２ ) ｍ -C(=O)R ７ , or -NH-(CH ２ CH ２ O) ｍ -CH ２ CH ２ -C(=O)R ７ And, Each m independently represents an integer of 1, 2, 3, 4, or 5. R ７ is OH or NR ８ａ R ８ｂ And, R ８ａ and R ８ｂ Each of these independently consists of H, substituted or unsubstituted C. １～４ Alkyl, substituted, or unsubstituted C ３～５ It is cycloalkyl, or R ８ａ and R ８ｂ These, together with the atoms to which they bond, become substituted or unsubstituted C ３～５ A compound according to any one of claims 71 to 86, which forms a cycloalkyl group.

88. R ７ However, OH, NH ２ , NHCH ３ , or N(CH ３ ) ２ The compound according to claim 87.

89. The compound according to any one of claims 71 to 88, wherein the subscript a' is 0.

90. The compound according to any one of claims 71 to 88, wherein the subscript a' is 1.

91. The aforementioned compound, 【Chemistry 75】 The compound according to claim 88, or a pharmaceutically acceptable salt, tautomer, or solvate thereof.

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