Immunomodulatory purine derivative compounds, complexes thereof, and methods of use thereof

Abstract

There is disclosed a purine derivative compound of general formula (I) (wherein X is O or NH; Y is CH or N; R 1 is optionally substituted alkyl; n is from 1 to 4; m is from 0 to 4; R 2 is H, halogen, alkyl or alkoxy; R 3 and R 1 are each H, OH, NR 6 R 7 optionally substituted alkyl, alkenyl, cycloalkyl, heterocycloalkyl, aryl or heteroaryl, or R 3 and R 4 and N may together form a ring). Also disclosed are immunopotentiating antibody conjugates (ISAC) containing such compounds, and methods for producing and using such compounds for the treatment of diseases such as cancer, for example. TIFF2025524965000338.tif48164

Description

Technical Field

[0001] The present disclosure relates to the field of therapeutic agents, and in particular, to purine analogs, complexes containing such purine derivative compounds, their use for modulating an immune response in cells or subjects, and the use of such compounds and complexes for the treatment of diseases (such as cancer, inflammation, etc.).

Background Art

[0002] Immunoactivating compounds and immunocomplexes containing such compounds (such as antibody-drug conjugates (ADCs) such as immune-stimulating antibody complexes (ISAC)) have previously been evaluated for the treatment of various diseases including cancer. However, such compounds and complexes are still not clinically proven mainly due to dose-limiting cytotoxicity (such as cytokine release syndrome (CRS)) that may limit their use to doses where sufficient efficacy cannot be obtained.

[0003] Furthermore, current immunotherapies lack efficacy, especially in solid tumors, due to the highly suppressive tumor microenvironment (TME) and the lack of associated effector immune cells (such as T cells). Compounds that stimulate toll-like receptor 7 (TLR7) are immunoactivating compounds that can locally "reprogram" the TME at the tumor site, but significant tolerability issues that limit the systemic administration of these compounds remain unresolved.

[0004] Clinical data support the use of TLR7 agonists in cancer treatment, although such treatments require local administration. For example, imiquimod, a TLR7 agonist, is approved for topical use in the treatment of skin tumors, including basal cell carcinoma and actinic keratosis (see Geisse et al. J Am Acad Dermatol. 2004;50(5):722-33, and Korman et al. Arch Dermatol. 2005;141(4):467-473). Other uses in invasive skin cancers (e.g., squamous cell carcinoma, Bowen's disease, melanoma, and / or malignant lentigo) are similarly effective when locally applied to surface lesions (see Meyer et al. Expert Opin Investig Drugs. 2008;17(7):1051-65, and Wolf et al. Arch Dermatol. 2003;139(3):273-6).

[0005] Unfortunately, studies have reported that systemic administration of imiquimod and other TLR agonists leads to dose-limiting toxicity at sub-effective doses (see Dudek et al. Clin Cancer Res 2007;13:7119-7125). Furthermore, overexpression and / or activation of some TLRs, including TLR7, can result in opposing anti-tumor / tumor-promoting activities, and in some cases, contribute to, rather than reduce, inflammation, tumor growth, cell survival, metastasis, and upregulation of inflammatory cytokines (see Kaczanowska S, et al. J Leukoc Biol. 2013;93(6):847-863). SUMMARY OF THE INVENTION

[0006] Purine derivative compounds, such as compounds having a structure according to any one of formulas (I)-(IV) herein, and complexes containing such compounds, such as complexes according to formula (X), are disclosed herein. Further, the present disclosure discloses methods for producing the compounds and complexes described herein, and their use, for example, for the treatment of diseases such as cancer.

[0007] The in vitro agonism of TLR7 using the compounds and complexes of the present disclosure was demonstrated by incubating immune cells expressing TLR7 (e.g., PBMC) with the compounds and complexes of the present disclosure and measuring one or more downstream effects of TLR7 agonism, such as cytokine induction (see, e.g., Example 3). Further, an in vivo immunostimulatory antibody-drug conjugate (ISAC) therapy utilizing an antibody targeting a tumor-associated antigen (e.g., Her2) conjugated to at least one compound of the present disclosure can elicit a potent anti-tumor response evidenced by suppression of tumor growth rate and / or reduction of tumor volume (see, e.g., Example 6). Surprisingly, a marked decrease in off-target effects, and thus improved tolerability, was confirmed during in vivo testing of certain ISACs comprising the TLR7-stimulating compounds of the present disclosure when compared to the use of conventional immunostimulatory compounds (see, e.g., Example 7).

[0008] More specifically, in various embodiments, the present disclosure relates to a compound of formula (I):

Chemical formula

Chemical formula

[0009] In some embodiments, the compound of formula (I) has the structure according to formula (II):

Chemical formula

Chemical formula

[0010] In various embodiments, the present disclosure relates to a compound of formula (III):

Chemical formula

Chemical formula

[0011] In certain embodiments of the present disclosure, the compound according to formula (III) has the formula (IV):

Chemical formula

Chemical formula

[0012] In some embodiments, the disclosure relates to a method of stimulating TLR7, the method comprising contacting a cell expressing TLR7 with a compound of any one of formulas (I)-(IV), thereby stimulating TLR7.

[0013] In some embodiments, the disclosure relates to a method of inducing cytokine release from a cell expressing TLR7, the method comprising contacting the cell with a compound of any one of formulas (I)-(IV), thereby inducing cytokine release from the cell.

[0014] In some embodiments, the present disclosure relates to a method of inhibiting the growth of cancer cells, the method comprising contacting a cell population comprising cancer cells and immune cells expressing TLR7 with an effective amount of any one of the compounds of formulas (I)-(IV). Such a method may further comprise stimulating TLR7 with any one of the compounds of formulas (I)-(IV), thereby inhibiting the growth of cancer cells.

[0015] In some embodiments, the present disclosure relates to a method of killing cancer cells, the method comprising contacting a cell population comprising cancer cells and immune cells expressing TLR7 with an effective amount of any one of the compounds of formulas (I)-(IV). Such a method may further comprise stimulating TLR7 with any one of the compounds of formulas (I)-(IV), thereby killing cancer cells.

[0016] Other embodiments of the present disclosure relate to a method of stimulating an immune response in a subject in need thereof, the method comprising administering to the subject an effective amount of any one of the compounds of formulas (I)-(IV). In some embodiments, the compound stimulates TLR7 in the subject, thereby stimulating an immune response in the subject.

[0017] Another embodiment of the present disclosure relates to a method of treating cancer in a subject in need thereof, the method comprising administering to the subject an effective amount of any one of the compounds of formulas (I)-(IV). Such a method may further comprise stimulating TLR7 in the subject, thereby further treating cancer in the subject.

[0018] In various embodiments, the present disclosure relates to a complex comprising one or more of any one of the compounds of formulas (I)-(IV). In various embodiments, such a complex has the structure according to formula (X): (X) T-[L-(C) p r having the structure according to (wherein T is a targeting moiety,​ L is a linker; C is a compound of any one of formulas (I)-(IV); p is an integer from 1 to 5; r has a value of from about 1 to about 8).

[0019] In various embodiments, the targeting moiety is an antibody or an antigen-binding fragment thereof.

[0020] The present disclosure also relates to a method of stimulating TLR7, the method comprising contacting a cell expressing TLR7 with a complex of formula (X), thereby stimulating TLR7.

[0021] Another embodiment herein relates to a method of inducing cytokine release from cells expressing TLR7, the method comprising contacting the cells with a complex of formula (X), thereby inducing cytokine release from the cells.

[0022] Another embodiment herein relates to a method of inhibiting cancer cell growth, the method comprising contacting a cell population comprising cancer cells and immune cells expressing TLR7 with an effective amount of a complex of formula (X). Such a method may further comprise stimulating TLR7 with a compound of the complex of formula (X), thereby inhibiting cancer cell growth.

[0023] Another embodiment of the present disclosure relates to a method of killing cancer cells, the method comprising contacting a cell population comprising cancer cells and immune cells expressing TLR7 with an effective amount of a complex of formula (X). Such a method may further comprise stimulating TLR7 with a compound of the complex of formula (X) (e.g., a compound of any one of formulas (I)-(IV) included in the complex), thereby killing the cancer cells.

[0024] Another embodiment of the present specification relates to a method for stimulating an immune response in a subject in need of stimulation of the immune response, the method comprising administering to the subject an effective amount of a complex of formula (X). In some embodiments, the complex stimulates TLR7 in the subject, for example, by a compound of formula (I)-(IV), thereby stimulating an immune response in the subject.

[0025] Another embodiment of the present disclosure relates to a method for treating cancer in a subject in need of treatment of cancer, the method comprising administering to the subject an effective amount of a complex of formula (X). In some embodiments, the complex stimulates TLR7 in the subject, for example, by a compound of formula (I)-(IV), thereby treating cancer in the subject.

[0026] The embodiments disclosed herein are shown by way of example and not by way of limitation in the accompanying drawings. The present specification and the drawings are for illustrative purposes only and are merely aids to understanding, and are not intended as definitions of limitations of the compounds, complexes and methods of the present disclosure.

Brief Description of the Drawings

[0027]

Figure 1

[0028]

Figure 2

[0029]

Figure 3

[0030]

Figure 4A

Figure 4B

Figure 4C

Figure 4D

Figure 4E

Figure 4F

Figure 4G

[0031]

Figure 5

Mode for Carrying Out the Invention

[0032] In various embodiments, the present disclosure relates to an immunostimulatory purine derivative compound having a structure according to any one of formulas (I)-(IV) that can stimulate TLR7. Also disclosed herein are compound-linker constructs of formula (A) comprising a compound of the present disclosure attached to a linker moiety. For example, by using the compound-linker constructs of formula (A) described herein, an immunostimulatory complex of formula (X) comprising one or more purine derivative compounds (e.g., compounds of formulas (I)-(IV)) described herein attached to a targeting moiety via a linker is further disclosed herein. For example, methods for generating and using the compounds, compound-linker constructs, and complexes for the treatment of cancer are also disclosed.

[0033] Definitions Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art.

[0034] The term "about", as used herein in connection with a numerical value or range, generally refers to ±10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, or 1% of the recited or claimed numerical value or range, unless otherwise specified. In various embodiments, the term "about" refers to a variation of approximately ±10% from a given value. In other embodiments, the term "about" refers to a variation of approximately ±5% from a given value. In yet another embodiment, the term "about" refers to a variation of approximately ±1% from a given value. It is to be understood that such variations are always included in any given value described herein, whether or not specifically recited.

[0035] The use of the word "a" or "an", as used herein in conjunction with the term "comprising", may mean "one" in some cases, but is also consistent with the meaning of "one or more", "at least one", and "one or more than one".

[0036] As used herein, the terms "comprising," "having," "including," and "containing," and grammatical variations thereof, are inclusive and do not exclude additional elements and / or method steps not recited. The term "consisting essentially of," when used herein in connection with a compound, composition, use, or method, indicates that additional elements and / or method steps may be present, but that these additional elements and / or method steps do not substantially affect the manner in which the recited compound, composition, method, or use functions. The term "consisting of," when used herein in connection with a compound, composition, use, or method, excludes the presence of additional elements and / or method steps. Compounds, compositions, uses, or methods described herein as including certain elements and / or steps may also, in certain embodiments, consist essentially of those elements and / or steps, and in other embodiments, consist of those elements and / or steps, whether or not these embodiments are expressly recited.

[0037] The terms "alkyloxycarbonyl" and "alkoxycarbonyl" can be used interchangeably herein and refer to a group -C(O)OR where R is alkyl.

[0038] As used herein, the term "alkyl" refers to a straight or branched saturated hydrocarbon group containing a specified number of carbon atoms. Examples of alkyl include, but are not limited to, methyl, ethyl, n - propyl, isopropyl, n - butyl, sec - butyl, isobutyl, t - butyl, pentyl, isopentyl, t - pentyl, neopentyl, 1 - methylbutyl, 2 - methylbutyl, n - hexyl, and the like.

[0039] As used herein, the term "alkylcycloalkyl" refers to an alkyl group as defined herein substituted with one cycloalkyl group as defined herein. Examples include (C1-C6 alkyl)-cycloalkyl, such as (C1-C2 alkyl)-cycloalkyl or (C1-C4 alkyl)-cycloalkyl. Thus, as used herein, the terms "alkylheterocycloalkyl", "alkylaryl", and "alkylheteroaryl" each refer to an alkyl group as defined herein substituted with one heterocycloalkyl group, aryl group, or heteroaryl group as further defined herein. Examples include (C1-C6 alkyl)-heterocycloalkyl, (C1-C6 alkyl)-aryl, and (C1-C6 alkyl)-heteroaryl. In various embodiments, any cycloalkyl, heterocycloalkyl, aryl, or heteroaryl group itself may be substituted with one or more of -NH2, -CO2H, -OH, carbonyl, halogen, C1-C2 alkyl, C1-C2 hydroxyalkyl, (C0-C2 alkyl)-cycloalkyl, (C0-C2 alkyl)-heterocycloalkyl, (C0-C2 alkyl)-aryl, and (C0-C2 alkyl)-heteroaryl as described herein.

[0040] As used herein, the term "amide" refers to the group -C(O)NRR', where R and R' are independently hydrogen, alkyl, aryl, heteroaryl, cycloalkyl, or heterocycloalkyl.

[0041] As used herein, the term "amino" refers to the group -NRR', where R and R' are independently hydrogen, alkyl, aryl, heteroaryl, cycloalkyl, or heterocycloalkyl.

[0042] The term "carboxy", as used herein, refers to the group -C(O)OR, where R is H, alkyl, aryl, heteroaryl, cycloalkyl or heterocycloalkyl.

[0043] The term "haloalkyl", as used herein, refers to an alkyl group as defined herein substituted with one or more halogen atoms.

[0044] The terms "halogen" and "halo", as used herein, refer to fluorine (F), bromine (Br), chlorine (Cl) and iodine (I).

[0045] The term "aminoalkyl", as used herein, refers to an alkyl group as defined herein substituted with one or more amino groups, for example, 1, 2 or 3 amino groups.

[0046] The term "aminoaryl", as used herein, refers to an aryl group as defined herein substituted with one amino group.

[0047] The term "aryl", as used herein, unless otherwise defined, refers to a monocyclic or bicyclic hydrocarbon ring system of 6 to 12 members in which at least one ring is aromatic. Examples of aryl include, but are not limited to, phenyl, naphthalenyl, 1,2,3,4 - tetrahydro - naphthalenyl, 5,6,7,8 - tetrahydro - naphthalenyl, indanyl, etc.

[0048] The term "cycloalkyl", as used herein, unless otherwise defined, refers to a monocyclic or bicyclic saturated hydrocarbon containing a specific number of carbon atoms. Examples of cycloalkyl include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptane, bicyclo[1.1.1]pentane, bicyclo[2.2.1]heptane, bicyclo[3.1.1]heptane, etc.

[0049] As used herein, the term "heteroaryl", unless otherwise defined, refers to a 5- to 12-membered monocyclic or bicyclic ring system containing a specific number of carbon atoms, having at least one ring atom being a heteroatom, and having at least one ring being aromatic (e.g., including C3-C7, such as imidazole, thiazole, etc.). Examples of heteroatoms include, but are not limited to, O, S, and N. Examples of heteroaryl include, but are not limited to, pyridyl, benzofuranyl, pyrazinyl, pyridazinyl, pyrimidinyl, triazinyl, quinolinyl, benzoxazolyl, benzothiazolyl, isoquinolinyl, quinazolinyl, quinoxalinyl, pyrrolyl, indolyl, etc.

[0050] As used herein, the term "heterocycloalkyl" refers to a monocyclic or bicyclic non-aromatic ring system containing a specific number of carbon atoms and having at least one ring atom being a heteroatom, such as O, S, or N (e.g., including C2-C6, such as aziridine, piperazine, etc.). A heterocyclyl substituent can be attached via any available ring atom, such as a ring carbon or a ring nitrogen. Examples of heterocycloalkyl include, but are not limited to, aziridinyl, azetidinyl, piperidinyl, morpholinyl, piperazinyl, pyrrolidinyl, etc.

[0051] As used herein with respect to an alkyl or heteroalkyl ring system, the term "bicyclic" generally includes both fused and spiro ring systems unless otherwise defined herein.

[0052] As used herein, the terms "hydroxy" and "hydroxyl" refer to the group -OH.

[0053] As used herein, the term "hydroxyalkyl" refers to a straight or branched alkyl group as defined herein substituted with one or more hydroxy groups. In some embodiments, such one or more hydroxy groups may be terminal hydroxy groups.

[0054] As used herein, the term "alkylthio" refers to the group -SR, where R is a C1-C6 alkyl group. In some embodiments, the alkyl group may be optionally substituted with one or more of -NH2, -CO2H, -OH, carbonyl, halogen, C1-C2 alkyl, and C1-C2 hydroxyalkyl.

[0055] As used herein, the terms "thio" and "thiol" refer to the group -SH.

[0056] Unless otherwise specifically stated as "unsubstituted", any alkyl (e.g., any haloalkyl, hydroxyalkyl, aminoalkyl, etc.), cycloalkyl, heterocycloalkyl, spirocycloalkyl, heterospirocycloalkyl, aryl, or heteroaryl group referred to herein is understood to be "optionally substituted", i.e., each such reference includes both unsubstituted and substituted forms of these groups. For example, a reference to a "-C1-C6 alkyl" group includes both unsubstituted -C1-C6 alkyl and C1-C6 alkyl substituted with one or more substituents further described herein.

[0057] In various embodiments, any one of the substituents of Formulas (I)-(IV) in any of the embodiments herein may be optionally substituted with one or more groups selected from: -NH2, -CO2H, -OH, carbonyl, halogen, C1-C4 alkyl, C1-C4 hydroxyalkyl, C1-C4 aminoalkyl, C1-C4 carboxyalkyl, (C0-C4 alkyl)-cycloalkyl, (C0-C4 alkyl)-heterocycloalkyl, (C0-C4 alkyl)-spirocycloalkyl, (C0-C4 alkyl)-heterospirocycloalkyl, (C0-C4 alkyl)-aryl, and (C0-C4 alkyl)-heteroaryl, wherein the alkyl, hydroxyalkyl, C1-C4 aminoalkyl, C1-C4 carboxyalkyl, cycloalkyl, heterocycloalkyl, spirocycloalkyl, heterospirocycloalkyl, aryl or heteroaryl group may itself be substituted with one or more of -NH2, -CO2H, -OH, carbonyl, halogen, unsubstituted C1-C4 alkyl, unsubstituted C1-C4 hydroxyalkyl, unsubstituted C1-C4 aminoalkyl, or unsubstituted C1-C4 carboxyalkyl.

[0058] In other embodiments, any one of the substituents of Formulas (I)-(IV) in any of the embodiments herein may be optionally substituted with one or more groups selected from: -NH2, -CO2H, -OH, carbonyl, halogen, unsubstituted C1-C4 alkyl, unsubstituted C1-C4 hydroxyalkyl, unsubstituted C1-C4 aminoalkyl, unsubstituted C1-C4 carboxyalkyl, unsubstituted (C0-C4 alkyl)-cycloalkyl, unsubstituted (C0-C2 alkyl)-heterocycloalkyl, unsubstituted (C0-C2 alkyl)-aryl, and unsubstituted (C0-C2 alkyl)-heteroaryl.

[0059] In yet another embodiment, any one of the substituents of Formulas (I)-(IV) in any of the embodiments herein may be optionally substituted with one or more groups selected from the following: -NH2, -CO2H, -OH, carbonyl, halogen, unsubstituted C1-C4 alkyl, unsubstituted C1-C4 hydroxyalkyl, unsubstituted C1-C4 aminoalkyl, and unsubstituted C1-C4 carboxyalkyl.

[0060] Generally, a "substituted" substituent of any one of Formulas (I)-(IV) described herein may include one substituent, or a plurality of substituents up to the maximum valence of substitution of that group. For example, a methyl group can include 1, 2, or 3 substituents, and a phenyl group can include 1, 2, 3, 4, or 5 substituents. When a group is substituted with two or more substituents, the substituents may be the same or different.

[0061] Throughout this disclosure, unless otherwise specified, the following system was used for numbering the ring atoms in the purine moiety of the compounds described herein (

Chemical formula

Chemical formula

[0062] The terms "subject" and "patient" can be used interchangeably herein and refer to an animal in need of treatment. The animal in need of treatment may be a human or non-human animal, such as a mammal, bird or fish. In certain embodiments, the subject or patient is a mammal. In some embodiments, the subject is a human. In other embodiments, the subject is a rodent or non-human primate.

[0063] The "effective amount" of a compound or complex described herein with respect to a particular result to be achieved is an amount sufficient to achieve the desired result. For example, the "effective amount" of a compound when referred to in relation to killing cancer cells refers to the amount of that compound sufficient to bring about a killing effect.

[0064] It should be further understood that the affirmative listing of a feature in one embodiment serves as a basis for excluding that feature in alternative embodiments. In particular, when a list of options is presented for a given embodiment or claim, one or more of the options may be removed, and the shortened list may form an alternative embodiment, whether or not such an alternative embodiment is explicitly recited.

[0065] Any embodiment discussed herein is contemplated to be implementable with respect to any method, use, or composition disclosed herein, and vice versa.

[0066] The compounds of the present disclosure In various embodiments, the present disclosure discloses purine derivative compounds capable of stimulating TLR7.

[0067] In some embodiments, the present disclosure relates to formula (I): [Chemical formula] and relates to compounds having, their tautomers, protected forms, and / or pharmaceutically acceptable salts (wherein, X is O or NH; Y is N or CH; R 1 is optionally substituted C2-C6 alkyl or branched optionally substituted C3-C8 hydroxyalkyl; R 2 is H, halogen, optionally substituted C1-C6 alkyl, or optionally substituted C1-C6 alkoxy; R 3 and R 4is, independently, H, Q-R 5 or R 3 and R 4 together with N optionally form a C2-C6-heterocycloalkyl ring, an optionally substituted C3-C7-heterobicycloalkyl ring, an optionally substituted C6-C 10 -heterospirocycloalkyl ring or an optionally substituted heteroaryl ring; Q is a bond, an optionally substituted C1-C6-alkyl, an optionally substituted C2-C6-alkenyl or

Chemical formula

[0068] In some embodiments, in the compound of formula (I), R 2 is halogen, optionally substituted C1-C6 alkyl, or optionally substituted C1-C6 alkoxy. In some of these embodiments, R 2 is halogen or optionally substituted C1-C6 alkoxy. In certain embodiments, R 2 is halogen or unsubstituted C1-C6 alkoxy. In some embodiments, R 2 may be fluorine or methoxy. In some embodiments, R 2 is fluorine. In other embodiments, R 2 is methoxy.

[0069] Compounds of formula (I) wherein X is O are disclosed herein. In other embodiments, in the compound of formula (I), X is NH.

[0070] In some embodiments, in the compound of formula (I), R 1 is optionally substituted C2-C6 alkyl. In some embodiments, R1 is an unsubstituted C2-C6 alkyl, for example, an unsubstituted C2-C4 alkyl. In these embodiments, X may be O.

[0071] In yet another embodiment, R 1 is a branched, optionally substituted C3-C8 hydroxyalkyl. In some embodiments, R 1 may be a branched, unsubstituted C3-C8 hydroxyalkyl. In some embodiments, R 1 is a branched, unsubstituted C4-C6 hydroxyalkyl.

[0072] In such embodiments, the compound of formula (I) may be

Chemical formula

[0073] In some embodiments, in the compound of formula (I), R 3 and R 4 are independently H or Q-R 5 . In some embodiments, R 3 and R 4 may both be H. In other embodiments, R 3 and R 4 are both Q-R 5 . R 3 and R4 are both Q-R 5 In some embodiments where is, Q may be an optionally substituted C1-C6-alkyl, and R 5 may be H or OH. R 3 and R 4 are both Q-R 5 In some embodiments where is, Q is an unsubstituted C1-C6-alkyl, and R 5 is H.

[0074] In other embodiments, in the compound of formula (I), Q is a bond, and R 5 is an unsubstituted C3-C6-carboxyalkyl.

[0075] In various other embodiments, in the compound of formula (I), Q is a bond, optionally substituted C1-C6-alkyl, optionally substituted C2-C6-alkenyl, or

Chemical formula

[0076] R 3 and R 4 are both H. In some embodiments, the compound of formula (I) may be the following.

Chemical formula

[0077] R 3 and R 4 are both Q-R 5 In some embodiments, the compound of formula (I) may be the following.

Chemical formula

[0078] In yet another embodiment, in the compound of formula (I), R 3 is H and R 4 is Q-R 5 In such embodiments, Q may be a bond and R 5 may be optionally substituted C3-C6-carboxyalkyl, optionally substituted C3-C6-cycloalkyl, optionally substituted C2-C6-heterocycloalkyl, optionally substituted C3-C7-heterobicycloalkyl, optionally substituted C6-C 10 -heterospirocycloalkyl, optionally substituted aryl, or optionally substituted heteroaryl. In such embodiments, the compound of formula (I) may be selected from the compounds shown in Subtable 1C.

[0079] In some embodiments, in the compound of formula (I), R 3 is H, and R 4 is Q-R 5 where Q is optionally substituted C1-C6-alkyl and R 5 is H, OH, optionally substituted C3-C6-carboxyalkyl, optionally substituted C3-C6-cycloalkyl, optionally substituted C2-C6-heterocycloalkyl, optionally substituted C3-C7-heterobicycloalkyl, optionally substituted C6-C 10 -heterospirocycloalkyl, optionally substituted aryl, or optionally substituted heteroaryl. In some embodiments, Q is unsubstituted C1-C6-alkyl. In other embodiments, Q is C1-C6-alkyl substituted with one or more of -NH2, -CO2H, -OH, carbonyl, halogen, C1-C2 alkyl, C1-C2 hydroxyalkyl, (C0-C2 alkyl)-cycloalkyl, (C0-C2 alkyl)-heterocycloalkyl, (C0-C2 alkyl)-aryl, and (C0-C2 alkyl)-heteroaryl. In other embodiments, Q is C1-C6-alkyl substituted with one or more of -NH2, -CO2H, -OH, carbonyl, halogen, C1-C2 alkyl, and C1-C2 hydroxyalkyl. In some embodiments, the compound of formula (I) may be selected from the compounds shown in Sub-Table 1D.

[0080] In some embodiments, in the compound of formula (I), R 3 is H, and R 4 is Q-R 5 where Q is optionally substituted C2-C6-alkenyl and R 5 is H, OH, optionally substituted C3-C6-carboxyalkyl, optionally substituted C3-C6-cycloalkyl, optionally substituted C2-C6-heterocycloalkyl, optionally substituted C3-C7-heterobicycloalkyl, optionally substituted C6-C 10- Heterospirocycloalkyl, optionally substituted aryl, or optionally substituted heteroaryl. In some embodiments, Q is unsubstituted C2-C6-alkenyl.

[0081] In various embodiments, R 3 and R 4 One or more of is Q-R 5 In the compound of formula (I), Q is C3-C6-cycloalkyl, C2-C6-heterocycloalkyl, C3-C7-heterobicycloalkyl, C6-C 10 - Heterospirocycloalkyl, aryl, or C1-C6-alkyl substituted with one or more of heteroaryl. In some of these embodiments, such a ring substituent (referred to hereinafter as "W") may be a bridging moiety, whereby Q is

Chemical formula

[0082] In some of these embodiments, W is unsubstituted C3-C6-cycloalkyl, unsubstituted C2-C6-heterocycloalkyl, unsubstituted C3-C7-heterobicycloalkyl, unsubstituted C6-C 10 - Heterospirocycloalkyl, unsubstituted aryl, or unsubstituted heteroaryl. In such embodiments, Q is

Chemical formula

[0083] In some embodiments, in the compound of formula (I), R 3 and R 4 together with N form an optionally substituted C2-C6-heterocycloalkyl ring, an optionally substituted C3-C7-heterobicycloalkyl ring, an optionally substituted C6-C 10 -heterospirocycloalkyl ring or an optionally substituted heteroaryl ring. In some embodiments, R 3 and R 4 together with N form a C3-C6-heterocycloalkyl ring, a C3-C6-heterobicycloalkyl ring, a C6-C 10 -heterospirocycloalkyl ring, or a heteroaryl ring, and the ring moiety is optionally substituted with one or more of -NH2, -CO2H, -OH, carbonyl, halogen, alkylthio, C1-C2 alkyl, C1-C2 hydroxyalkyl, (C0-C2 alkyl)-cycloalkyl, and (C0-C2 alkyl)-heterocycloalkyl.

[0084] In some embodiments, R 3 and R 4 together with N

Chemical formula

[0085] In some embodiments, in the compound of formula (I), R 3 and R 4 together with N

Chemical formula

[0086] In some embodiments, in the compound of formula (I), R 3 and R 4 together with N form the ring moiety described herein, and the compound is selected from the compounds shown in Subtable 1B.

[0087] In some embodiments, at least one of R 3 and R 4 is Q-R 5 In the compound of formula (I), R 5 may be NR 6 R 7 In such embodiments, R 6 and R 7 may independently be H, optionally substituted C1-C6 alkyl, or optionally substituted C1-C6 alkoxycarbonyl. In some embodiments, R 6 and R 7 are both H. In some embodiments, R 6 and R 7 are both optionally substituted C1-C6 alkyl, and R 6 is the same as R 7 In such embodiments, R 6 and R 7 may both be C1-C6 alkyl substituted with one or more of -NH2, -CO2H, -OH, carbonyl, halogen, C1-C2 alkylthio, C1-C2 alkyl, and C1-C2 hydroxyalkyl. In some embodiments, R 6 and R 7 are C1-C2 alkyl substituted with one or more of -NH2, -CO2H, -OH, or halogen. In other embodiments, R 6 is H and R 7 is optionally substituted C1-C6 alkyl. In some embodiments, R 6 is H and R 7may be C1-C6 alkyl substituted with one or more of -NH2, -CO2H, -OH, carbonyl, halogen, C1-C2 alkylthio, C1-C2 alkyl, C1-C2 hydroxyalkyl, optionally substituted C3-C6-cycloalkyl, optionally substituted C2-C6-heterocycloalkyl, optionally substituted aryl, and optionally substituted heteroaryl.

[0088] In some embodiments, R 3 and R 4 at least one of which is Q-R 5 in the compound of formula (I), R 5 may be NR 6 R 7 and R 6 and R 7 together with N form an optionally substituted C2-C6-heterocycloalkyl ring, an optionally substituted C3-C7-heterobicycloalkyl ring, an optionally substituted C6-C 10 -heterospirocycloalkyl ring or an optionally substituted heteroaryl ring. In some embodiments, such ring moieties may be optionally substituted with one or more of -NH2, -CO2H, -OH, carbonyl, halogen, C1-C2 alkylthio, C1-C2 alkyl, C1-C3 aminoalkyl and C1-C2 hydroxyalkyl. In some embodiments, R 6 and R 7 together with N

Chemical formula

[0089] In some embodiments, the compound of formula (I) is selected from the compounds shown in Subtable 1E.

[0090] In some embodiments, in the compound of formula (I), n is 1.

[0091] In some embodiments, in the compound of formula (I), m is an integer from 0 to 3 or from 1 to 3. In some embodiments, m is 0. In some embodiments, m is 1. In some embodiments, m is 2. In some embodiments, m is 3.

[0092] In some embodiments, in the compound of formula (I), R 2 is H, (a) R 1 is a branched optionally substituted C3-C8 hydroxyalkyl, and / or (b) X is NH, R 1 is an optionally substituted C5-C6 alkyl, and / or (c) m is 0 or 1, R 3 and R 4 together with N form an unsubstituted piperazinyl ring or an optionally substituted C6-C 10 -heterospirocycloalkyl ring. In some embodiments, R 2 is H, R 1 is a branched optionally substituted C3-C8 hydroxyalkyl. In such embodiments, R 1 may be a branched unsubstituted C3-C8 hydroxyalkyl. In other embodiments, R 2 is H, X is NH, R 1 is an optionally substituted C5-C6 alkyl. In some of those embodiments, R 1 is an unsubstituted C5-C6 alkyl. In yet another embodiment, R 2 may be H, m may be 0 or 1, R 3 and R 4 together with N may form an unsubstituted piperazinyl ring, or an optionally substituted C6-C 10 -heterospirocycloalkyl ring. In some embodiments, R 3 and R 4 together with N form an unsubstituted piperazinyl ring. In yet another embodiment, R 3 and R 4 together with N may form an optionally substituted C6-C 10- forms a heterospirocycloalkyl ring.

[0093] In some embodiments, the compound of formula (I) is selected from Table 1 herein, including Sub-tables 1A - 1F.

[0094] In certain embodiments of the present disclosure, the compound of formula (I) has the formula (II): [Chemical formula] having the structure of (wherein, X is O or NH; Y is N or CH; R 1 is optionally substituted C2 - C6 alkyl or branched optionally substituted C3 - C8 hydroxyalkyl; R 3 and R 4 are independently H, Q - R 5 or R 3 and R 4 together with N form an optionally substituted C2 - C6 - heterocycloalkyl ring, an optionally substituted C3 - C7 - heterobicycloalkyl ring, an optionally substituted C6 - C 10 - heterospirocycloalkyl ring or an optionally substituted heteroaryl ring; Q is a bond, optionally substituted C1 - C6 - alkyl, optionally substituted C2 - C6 - alkenyl or [Chemical formula] wherein, * is the point of attachment to R 5 and # is the point of attachment to N; R 5 is H, OH, NR 6 R 7 , optionally substituted C3 - C6 - carboxyalkyl, optionally substituted C3 - C6 - cycloalkyl, optionally substituted C2 - C6 - heterocycloalkyl, optionally substituted C3 - C7 - heterobicycloalkyl, optionally substituted C6 - C 10-heterospirocycloalkyl, optionally substituted aryl, or optionally substituted heteroaryl; R 6 and R 7 are independently H, optionally substituted C1-C6 alkyl, optionally substituted C1-C6 alkoxycarbonyl, or R 6 and R 7 together with N form an optionally substituted C2-C6 heterocycloalkyl ring, an optionally substituted C3-C7 heterobicycloalkyl ring, an optionally substituted C6-C 10 -heterospirocycloalkyl ring or an optionally substituted heteroaryl ring; W is optionally substituted C3-C6 cycloalkyl, optionally substituted C2-C6 heterocycloalkyl, optionally substituted C3-C7 heterobicycloalkyl, optionally substituted C6-C 10 -heterospirocycloalkyl, optionally substituted aryl, or optionally substituted heteroaryl; m is an integer from 0 to 4; (a and b are independently integers from 0 to 4).

[0095] In some embodiments, in the compound of formula (II), X is O and R 1 is optionally substituted C2-C6 alkyl. In some of these embodiments, R 1 is unsubstituted C2-C6 alkyl. In some specific embodiments, R 1 is ethyl or n-butyl.

[0096] In some embodiments, in the compound of formula (II), Y is CH.

[0097] In some embodiments, in the compound of formula (II), m is 0 or 1, 1 or 2, or 1 or 3. In such embodiments, m may be 0. In other embodiments, m is 1, 2, or 3. In some embodiments, m is 1. In some embodiments, m is 2. In yet another embodiment, m is 3.

[0098] In some embodiments, in the compound of formula (II), R 3 and R4 are both H. In other embodiments, R 3 and R 4 are both Q-R 5 . In such embodiments, Q may be optionally substituted C1-C6-alkyl, and R 5 may be H or OH. In some of these embodiments, Q is unsubstituted C1-C6-alkyl and R 5 is H. In other embodiments, Q is a bond and R 5 is unsubstituted C3-C6-carboxyalkyl.

[0099] In yet another embodiment, in the compound of formula (II), R 3 is H and R 4 is Q-R 5 . In such embodiments, Q may be a bond and R 5 may be optionally substituted C3-C6-carboxyalkyl, optionally substituted C3-C6-cycloalkyl, optionally substituted C2-C6-heterocycloalkyl, optionally substituted C3-C7-heterobicycloalkyl, optionally substituted C6-C 10 -heterospirocycloalkyl, optionally substituted aryl, or optionally substituted heteroaryl.

[0100] In some embodiments, in the compound of formula (II), R 3 is H and R 4 is Q-R 5wherein Q is optionally substituted C1-C6-alkyl, and R 5 is H, OH, optionally substituted C3-C6-carboxyalkyl, optionally substituted C3-C6-cycloalkyl, optionally substituted C2-C6-heterocycloalkyl, optionally substituted C3-C7-heterobicycloalkyl, optionally substituted C6-C 10 -heterospirocycloalkyl, optionally substituted aryl, or optionally substituted heteroaryl. In some embodiments, Q is unsubstituted C1-C6-alkyl. In other embodiments, Q is C1-C6-alkyl substituted with one or more of -NH2, -CO2H, -OH, carbonyl, halogen, C1-C2 alkylthio, C1-C2 alkyl, C1-C2 hydroxyalkyl, (C0-C2 alkyl)-cycloalkyl, (C0-C2 alkyl)-heterocycloalkyl, (C0-C2 alkyl)-aryl, and (C0-C2 alkyl)-heteroaryl. In other embodiments, Q is C1-C6-alkyl substituted with one or more of -NH2, -CO2H, -OH, carbonyl, halogen, C1-C2 alkyl, and C1-C2 hydroxyalkyl.

[0101] In some embodiments, in the compound of formula (II), R 3 is H, and R 4 is Q-R 5 wherein Q is optionally substituted C2-C6-alkenyl, and R 5 is H, OH, optionally substituted C3-C6-carboxyalkyl, optionally substituted C3-C6-cycloalkyl, optionally substituted C2-C6-heterocycloalkyl, optionally substituted C3-C7-heterobicycloalkyl, optionally substituted C6-C 10 -heterospirocycloalkyl, optionally substituted aryl, or optionally substituted heteroaryl. In some embodiments, Q is unsubstituted C2-C6-alkenyl.

[0102] In some of these embodiments, the compound is as follows.

Chemical Formula

[0103] In various embodiments, R 3 and R 4 In the compound of formula (II) in which one or more of are Q-R 5 Q is C1-C6-alkyl substituted with one or more of C3-C6-cycloalkyl, C2-C6-heterocycloalkyl, C3-C7-heterobicycloalkyl, C6-C 10 -heterospirocycloalkyl, aryl, or heteroaryl. In some of these embodiments, such ring substituents (referred to below as "W") may be a bridging moiety, whereby Q is [Chemical formula] wherein W is optionally substituted C3-C6-cycloalkyl, optionally substituted C2-C6-heterocycloalkyl, optionally substituted C3-C7-heterobicycloalkyl, optionally substituted C6-C 10 -heterospirocycloalkyl, optionally substituted aryl, or optionally substituted heteroaryl; a and b are independently integers from 0 to 4, * is the point of attachment to R 5 and # is the point of attachment to N.

[0104] In such embodiments, W is an unsubstituted ring moiety, whereby Q is [Chemical formula] may be selected from the group consisting of, wherein a and b are independently 1, 2, or 3.

[0105] In some embodiments, in the compound of formula (II), R 3 and R 4is a C2-C6-heterocycloalkyl ring optionally substituted together with N, a C3-C7-heterobicycloalkyl ring optionally substituted, a C6-C 10 -heterospirocycloalkyl ring or a heteroaryl ring optionally substituted. In some embodiments, R 3 and R 4 form, together with N, a C2-C6-heterocycloalkyl ring, a C3-C7-heterobicycloalkyl ring, a C6-C 10 -heterospirocycloalkyl ring, or a heteroaryl ring, and the ring moiety is optionally substituted with one or more of -NH2, -CO2H, -OH, carbonyl, halogen, alkylthio, C1-C2 alkylthio, C1-C2 alkyl, C1-C2 hydroxyalkyl, (C0-C2 alkyl)-cycloalkyl, or (C0-C2 alkyl)-heterocycloalkyl. In some embodiments, R 3 and R 4 form, together with N,

Chemical formula

[0106] In some embodiments, in the compound of formula (II), R 3 and R 4 form, together with N,

Chemical formula

[0107] In some embodiments, at least one of R 3 and R 4 is Q-R 5In the compound of formula (II), R 5 is NR 6 R 7 which may be. In such embodiments, R 6 and R 7 may each independently be H, optionally substituted C1-C6 alkyl, or optionally substituted C1-C6 alkoxycarbonyl. In some embodiments, R 6 and R 7 are both H. In some embodiments, R 6 and R 7 are both optionally substituted C1-C6 alkyl, whereby R 6 is the same as R 7 . In such embodiments, R 6 and R 7 may both be C1-C6 alkyl substituted with one or more of -NH2, -CO2H, -OH, carbonyl, halogen, C1-C2 alkylthio, C1-C2 alkyl, and C1-C2 hydroxyalkyl. In some embodiments, R 6 and R 7 are C1-C2 alkyl substituted with one or more of -NH2, -CO2H, -OH, and halogen. In other embodiments, R 6 is H and R 7 is optionally substituted C1-C6 alkyl. In some embodiments, R 6 is H and R 7 may be C1-C6 alkyl substituted with one or more of -NH2, -CO2H, -OH, carbonyl, halogen, C1-C2 alkylthio, C1-C2 alkyl, C1-C2 hydroxyalkyl, optionally substituted C3-C6-cycloalkyl, optionally substituted C2-C6-heterocycloalkyl, optionally substituted aryl, and optionally substituted heteroaryl.

[0108] In some embodiments, in the compound of formula (II), R 5 is NR 6 R 7 and R6 and R7 together with N are optionally substituted C2-C 6-Heterocycloalkyl ring, optionally substituted C 3 ~C7-Heterobicycloalkyl ring, optionally substituted C6~C 10 -Heterospirocycloalkyl ring or optionally substituted heteroaryl ring is formed. In some embodiments, such ring moieties may be optionally substituted with one or more of -NH2, -CO2H, -OH, carbonyl, halogen, C1~C2 alkylthio, C1~C2 alkyl, C1~C3 aminoalkyl and C1~C2 hydroxyalkyl. In some embodiments, R 6 and R 7 together with N

Chemical formula

[0109] In some embodiments, the compound of formula (II) is selected from Subtable 1A.

[0110] In some embodiments, the compound of formula (I) or formula (II) is as follows.

Chemical formula

[0111] In various embodiments, the present disclosure relates to a compound of formula (III):

Chemical formula

[0112] In some embodiments, in the compound of formula (III), R 1a is optionally substituted C2-C6 alkyl. In such embodiments, R 1a may be unsubstituted C2-C6 alkyl. In some of these embodiments, R 1a may be ethyl or n-butyl.

[0113] In some embodiments, in the compound of formula (III), R 2a is halogen or optionally substituted C1-C6 alkoxy. In such embodiments, R 2a may be halogen or unsubstituted C1-C6 alkoxy. In certain embodiments, R 2a is fluorine or methoxy.

[0114] In some embodiments, in the compound of formula (III), R 3a and R 4a are both H. In other embodiments, R 3a and R 4a are both Q-R 5a In such embodiments, Q may be optionally substituted C1-C6-alkyl, and R 5a may be H or OH. In some of these embodiments, Q is unsubstituted C1-C6-alkyl and R 5a is H. In other embodiments, Q is a bond and R 5a is unsubstituted C3-C6-carboxyalkyl.

[0115] In yet another embodiment, in the compound of formula (III), R 3a is H and R4a is Q-R 5a In such embodiments, Q may be a bond and R 5a may be optionally substituted C3-C6-carboxyalkyl, optionally substituted C3-C6-cycloalkyl, optionally substituted C2-C6-heterocycloalkyl, optionally substituted C3-C7-heterobicycloalkyl, optionally substituted C6-C 10 -heterospirocycloalkyl, optionally substituted aryl, or optionally substituted heteroaryl.

[0116] In some embodiments, in the compound of formula (III), R 3a is H and R 4a is Q-R 5a where Q is optionally substituted C1-C6-alkyl and R 5a is H, OH, optionally substituted C3-C6-carboxyalkyl, optionally substituted C3-C6-cycloalkyl, optionally substituted C2-C6-heterocycloalkyl, optionally substituted C3-C7-heterobicycloalkyl, optionally substituted C6-C 10 -heterospirocycloalkyl, optionally substituted aryl, or optionally substituted heteroaryl. In some embodiments, Q is unsubstituted C1-C6-alkyl. In other embodiments, Q is C1-C6-alkyl substituted with one or more of -NH2, -CO2H, -OH, carbonyl, halogen, C1-C2 alkylthio, C1-C2 alkyl, C1-C2 hydroxyalkyl, (C0-C2 alkyl)-cycloalkyl, (C0-C2 alkyl)-heterocycloalkyl, (C0-C2 alkyl)-aryl, and (C0-C2 alkyl)-heteroaryl. In other embodiments, Q is C1-C6-alkyl substituted with one or more of -NH2, -CO2H, -OH, carbonyl, halogen, C1-C2 alkyl and C1-C2 hydroxyalkyl.

[0117] In some embodiments, in the compound of formula (III), R 3a is H and R 4a is Q-R5a and Q is optionally substituted C2-C6-alkenyl, and R 5a is H, OH, optionally substituted C3-C6-carboxyalkyl, optionally substituted C3-C6-cycloalkyl, optionally substituted C2-C6-heterocycloalkyl, optionally substituted C3-C7-heterobicycloalkyl, optionally substituted C6-C 10 -heterospirocycloalkyl, optionally substituted aryl, or optionally substituted heteroaryl. In some embodiments, Q is unsubstituted C2-C6-alkenyl.

[0118] In various embodiments, R 3a and R 4a one or more of which is Q-R 5a in the compound of formula (III), Q is C1-C6-alkyl substituted with one or more of C3-C6-cycloalkyl, C2-C6-heterocycloalkyl, C3-C7-heterobicycloalkyl, C6-C 10 -heterospirocycloalkyl, aryl, or heteroaryl. In some of these embodiments, such ring substituents (hereinafter referred to as "W") may be bridging moieties, whereby Q is [Chemical formula] wherein W is optionally substituted C3-C6-cycloalkyl, optionally substituted C2-C6-heterocycloalkyl, optionally substituted C3-C7-heterobicycloalkyl, optionally substituted C6-C 10 -heterospirocycloalkyl, optionally substituted aryl, or optionally substituted heteroaryl; a and b are independently integers from 0 to 4, * is the point of attachment to R 5 and # is the point of attachment to N.

[0119] In some of these embodiments, W is an unsubstituted ring moiety, whereby Q is [Chemical formula] may be selected from the group consisting of, wherein a and b are independently 1, 2, or 3.

[0120] In some embodiments, in the compound of formula (III), R 3a and R 4a together with N optionally form a C2-C6-heterocycloalkyl ring, an optionally substituted C3-C7-heterobicycloalkyl ring, an optionally substituted C6-C 10 -heterospirocycloalkyl ring or an optionally substituted heteroaryl ring. In some embodiments, R 3a and R 4a together with N form a C2-C6-heterocycloalkyl ring, a C3-C7-heterobicycloalkyl ring, a C6-C 10 -heterospirocycloalkyl ring, or a heteroaryl ring, and the ring moiety may be optionally substituted with one or more of -NH2, -CO2H, -OH, carbonyl, halogen, C1-C2 alkylthio, C1-C2 alkyl, C1-C2 hydroxyalkyl, (C0-C2 alkyl)-cycloalkyl, and (C0-C2 alkyl)-heterocycloalkyl.

[0121] In some embodiments, R 3a and R 4a together with N

Chemical formula

[0122] In some embodiments, in the compound of formula (III), R 3a and R 4a together with N [Chemistry] forms a ring selected from the group consisting of.

[0123] In some embodiments, in the compound of formula (III), R 5a is NR 6a R 7a In such embodiments, R 6a and R 7a may each independently be H, optionally substituted C1-C6 alkyl, or optionally substituted C1-C6 alkoxycarbonyl. In some embodiments, R 6a and R 7a are both H. In other embodiments, R 6a and R 7a are both optionally substituted C1-C6 alkyl, and R 6a is the same as R 7a In such embodiments, R 6a and R 7a may both be C1-C6 alkyl substituted with one or more of -NH2, -CO2H, -OH, carbonyl, halogen, C1-C2 alkylthio, C1-C2 alkyl, and C1-C2 hydroxyalkyl. In some embodiments, R 6a and R 7a are C1-C2 alkyl substituted with one or more of -NH2, -CO2H, -OH, or halogen. In other embodiments, R 6a is H and R 7a is optionally substituted C1-C6 alkyl. In some embodiments, R 6a is H and R 7a may be C1-C6 alkyl substituted with one or more of -NH2, -CO2H, -OH, carbonyl, halogen, C1-C2 alkylthio, C1-C2 alkyl, C1-C2 hydroxyalkyl, optionally substituted C3-C6-cycloalkyl, optionally substituted C2-C6-heterocycloalkyl, optionally substituted aryl, and optionally substituted heteroaryl.

[0124] In some embodiments, R 3a and R 4a in which at least one of 5a is Q-R 5a In the compound of formula (III), R 6a R 7a may be, and R 6a and R 7a together with N, optionally substituted C2-C6 heterocycloalkyl ring, optionally substituted C3-C7 heterobicycloalkyl ring, optionally substituted C6-C 10 -heterospirocycloalkyl ring or an optionally substituted heteroaryl ring. In some embodiments, such ring moieties may be optionally substituted with one or more of -NH2, -CO2H, -OH, carbonyl, halogen, C1-C2 alkylthio, C1-C2 alkyl, C1-C3 aminoalkyl and C1-C2 hydroxyalkyl. In some embodiments, R 6 and R 7 together with N

Chemical formula

[0125] In various embodiments, in the compound of formula (III), u is 1.

[0126] In various embodiments, in the compound of formula (III), v is an integer from 0 to 4, from 1 to 4, from 1 to 3 or from 1 to 2. In some embodiments, v is 0. In some embodiments, v is 1. In other embodiments, v is 2. In still other embodiments, v is 3.

[0127] In some embodiments, the compound of formula (III) is selected from Sub-Table 1A.

[0128] In various embodiments of the present disclosure, the compounds herein of formula (III) have the formula (IV):

Chemical formula

Chemical formula

[0129] In some embodiments, in the compound of formula (IV), R 1a is unsubstituted C2-C6 alkyl, for example, unsubstituted C2-C4 alkyl. In other embodiments, R 1a is branched unsubstituted C3-C8 hydroxyalkyl.

[0130] In some embodiments, in the compound of formula (IV), R 3a and R4a are both H. In other embodiments, R 3a and R 4a are both Q-R 5a . In such embodiments, Q may be optionally substituted C1-C6-alkyl, and R 5a may be H or OH. In some of these embodiments, Q is unsubstituted C1-C6-alkyl and R 5a is H. In other embodiments, Q is a bond and R 5a is unsubstituted C3-C6-carboxyalkyl.

[0131] In yet another embodiment, in the compound of formula (IV), R 3a is H and R 4a is Q-R 5a . In such embodiments, Q may be a bond and R 5ais H, an optionally substituted C3-C6-carboxyalkyl, an optionally substituted C3-C6-cycloalkyl, an optionally substituted C2-C6-heterocycloalkyl, an optionally substituted C3-C7-heterobicycloalkyl, an optionally substituted C6-C 10 -heterospirocycloalkyl, an optionally substituted aryl, or an optionally substituted heteroaryl.

[0132] In some embodiments, in the compound of formula (IV), R 3a is H, and R 4a is Q-R 5a where Q is an optionally substituted C1-C6-alkyl, and R 5a is H, OH, an optionally substituted C3-C6-carboxyalkyl, an optionally substituted C3-C6-cycloalkyl, an optionally substituted C2-C6-heterocycloalkyl, an optionally substituted C3-C7-heterobicycloalkyl, an optionally substituted C6-C 10 -heterospirocycloalkyl, an optionally substituted aryl, or an optionally substituted heteroaryl. In some embodiments, Q is an unsubstituted C1-C6-alkyl. In other embodiments, Q is a C1-C6-alkyl substituted with one or more of -NH2, -CO2H, -OH, carbonyl, halogen, C1-C2 alkylthio, C1-C2 alkyl, C1-C2 hydroxyalkyl, (C0-C2 alkyl)-cycloalkyl, (C0-C2 alkyl)-heterocycloalkyl, (C0-C2 alkyl)-aryl, and (C0-C2 alkyl)-heteroaryl. In yet another embodiment, Q is a C1-C6-alkyl substituted with one or more of -NH2, -CO2H, -OH, carbonyl, halogen, C1-C2 alkyl and C1-C2 hydroxyalkyl.

[0133] In some embodiments, in the compound of formula (IV), R 3a is H, and R 4a is Q-R 5a where Q is an optionally substituted C2-C6-alkenyl, and R 5ais H, OH, optionally substituted C3-C6-carboxyalkyl, optionally substituted C3-C6-cycloalkyl, optionally substituted C2-C6-heterocycloalkyl, optionally substituted C3-C7-heterobicycloalkyl, optionally substituted C6-C 10 -heterospirocycloalkyl, optionally substituted aryl, or optionally substituted heteroaryl. In some embodiments, Q is unsubstituted C2-C6-alkenyl.

[0134] In various embodiments, R 3a and R 4a one or more of which is Q-R 5a In the compound of formula (IV), Q is C1-C6-alkyl substituted with one or more of C3-C6-cycloalkyl, C2-C6-heterocycloalkyl, C3-C7-heterobicycloalkyl, C6-C 10 -heterospirocycloalkyl, aryl, or heteroaryl. In some of these embodiments, such a ring substituent (hereinafter referred to as "W") may be a bridging moiety, whereby Q is

Chemical formula

[0135] In such embodiments, W is an unsubstituted ring moiety, whereby Q is

Chemical formula

[0136] In some embodiments, in the compound of formula (IV), R 3a and R 4a together with N form an optionally substituted C2-C6-heterocycloalkyl ring, an optionally substituted C3-C7-heterobicycloalkyl ring, an optionally substituted C6-C 10 -heterospirocycloalkyl ring or an optionally substituted heteroaryl ring. In some embodiments, R 3a and R 4a together with N form a C2-C6-heterocycloalkyl ring, a C3-C7-heterobicycloalkyl ring, a C6-C 10 -heterospirocycloalkyl ring, or a heteroaryl ring, and the ring moiety may be optionally substituted with one or more of -NH2, -CO2H, -OH, carbonyl, halogen, C1-C2 alkylthio, C1-C2 alkyl, C1-C2 hydroxyalkyl, (C0-C2 alkyl)-cycloalkyl, or (C0-C2 alkyl)-heterocycloalkyl. In some embodiments, R 3a and R 4a together with N

Chemical formula

[0137] In some embodiments, in the compound of formula (IV), R 3a and R 4a together with N

Chemical formula

[0138] In some embodiments, one or more of R 3a and R 4a is Q-R 5a In the compound of formula (IV), R 5a may be NR 6a R 7a In such embodiments, R 6a and R 7a may independently be H, optionally substituted C1-C6 alkyl, or optionally substituted C1-C6 alkoxycarbonyl. In some embodiments, R 6a and R 7a are both H. In some embodiments, R 6a and R 7a are both optionally substituted C1-C6 alkyl, and R 6a is the same as R 7a In such embodiments, R 6a and R 7a may be C1-C6 alkyl substituted with one or more of -NH2, -CO2H, -OH, carbonyl, halogen, C1-C2 alkylthio, C1-C2 alkyl, and C1-C2 hydroxyalkyl. In some embodiments, R 6a and R 7a are C1-C2 alkyl substituted with one or more of -NH2, -CO2H, -OH, or halogen. In other embodiments, R 6a is H and R 7a is optionally substituted C1-C6 alkyl. In some embodiments, R 6a is H and R 7a may be C1-C6 alkyl substituted with one or more of -NH2, -CO2H, -OH, carbonyl, halogen, C1-C2 alkylthio, C1-C2 alkyl, C1-C2 hydroxyalkyl, optionally substituted C3-C6-cycloalkyl, optionally substituted C2-C6-heterocycloalkyl, optionally substituted aryl, and optionally substituted heteroaryl.

[0139] In some embodiments, R 3a and R 4a in one or more of the compounds of formula (IV) which is Q-R 5a , R 5a may be NR 6a R 7a , and R 6a and R 7a together with N form an optionally substituted C2-C6-heterocycloalkyl ring, an optionally substituted C3-C7-heterobicycloalkyl ring, an optionally substituted C6-C 10 -heterospirocycloalkyl ring or an optionally substituted heteroaryl ring. In some embodiments, such ring moieties may be optionally substituted with one or more of -NH2, -CO2H, -OH, carbonyl, halogen, C1-C2 alkylthio, C1-C2 alkyl, C1-C3 aminoalkyl and C1-C2 hydroxyalkyl. In some embodiments, R 6 and R 7 together with N

Chemical formula

[0140] In some embodiments, in the compounds of formula (IV), v is 0. In other embodiments, v is 1. In some embodiments, v is 2. In still other embodiments, v is 3.

[0141] In some embodiments, the compounds of formula (IV) are selected from the compounds shown in Sub-table 1A.

[0142] In some embodiments, the compounds of the present disclosure, for example, any one of the compounds of formulas (I)-(IV) herein, are selected from Table 1, including Sub-tables 1A-1F.

Table 1-1

Table 1-2

Table 1-3

Table 1-4

Table 1-5

Table 1-6

Table 1-7

Table 1-8

Table 1-9

Table 1-10

Table 1-11

Table 1-12

Table 1-13

Table 1-14

Table 1-15

Table 1-16

Table 1-17

Table 1-18

Table 1-19

Table 1-20

Table 1-21

Table 1-22

Table 1-23

Table 1-24

Table 1-25

Table 1-26

Table 1-27

Table 1-28

Table 1-29

Table 1-30

Table 1-31

Table 1-32

Table 1-33

Table 1-34

Table 1-35

Table 1-36

Table 1-37

[0143] Throughout the present disclosure, reference to a compound of formula (I) is understood in various embodiments to include compounds of formulas (I)-(IV) to the same extent as embodiments specifically listing each of these formulas individually.

[0144] In certain embodiments, the compounds having formula (I) described herein may have sufficiently acidic groups, sufficiently basic groups, or both functional groups, and thus may react with many organic and inorganic bases, or organic and inorganic acids, to form pharmaceutically acceptable salts. The term "pharmaceutically acceptable salts" as used herein refers to salts of compounds having formula (I) that are substantially non-toxic to organisms. Typical pharmaceutically acceptable salts include salts prepared by the reaction of a compound having formula (I) with a pharmaceutically acceptable inorganic or organic acid or organic or inorganic base. Such salts are known as acid addition salts and base addition salts.

[0145] Acids commonly used to form acid addition salts include, but are not limited to, inorganic acids such as hydrochloric acid, hydrobromic acid, hydroiodic acid, sulfuric acid, and phosphoric acid, as well as organic acids such as p-toluenesulfonic acid, methanesulfonic acid, oxalic acid, p-bromophenylsulfonic acid, carbonic acid, succinic acid, citric acid, benzoic acid, and acetic acid. Examples of pharmaceutically acceptable salts include, but are not limited to, sulfate, pyrosulfate, bisulfate, sulfite, phosphate, monohydrogen phosphate, dihydrogen phosphate, metaphosphate, pyrophosphate, bromide, iodide, acetate, propionate, decanoate, caprylate, acrylate, formate, hydrochloride, dihydrochloride, isobutyrate, caproate, heptanoate, propiolate, oxalate, malonate, succinate, suberate, sebacate, fumarate, maleate, butyne-1,4-dioate, hexyne-1,6-dioate, benzoate, chlorobenzoate, methylbenzoate, hydroxybenzoate, methoxybenzoate, phthalate, xylenesulfonate, phenylacetate, phenylpropionate, phenylbutyrate, citrate, lactate, gamma-hydroxybutyrate, glycolate, tartrate, methanesulfonate, propanesulfonate, naphthalene-1-sulfonate, naphthalene-2-sulfonate, and mandelate. Particularly targeted pharmaceutically acceptable acid addition salts are salts formed with inorganic acids such as hydrochloric acid and hydrobromic acid, as well as salts formed with organic acids such as maleic acid and methanesulfonic acid.

[0146] Salts of amine groups may also include quaternary ammonium salts in which the amino nitrogen has a suitable organic group such as an alkyl, lower alkenyl, lower alkynyl, or aralkyl moiety.

[0147] Examples of the base addition salts include salts derived from inorganic bases such as ammonium or alkali or alkaline earth metal hydroxides, carbonates, bicarbonates and the like. Examples of the bases useful for preparing pharmaceutically acceptable salts include, but are not limited to, sodium hydroxide, potassium hydroxide, ammonium hydroxide, potassium carbonate, sodium carbonate, sodium bicarbonate, potassium bicarbonate, calcium hydroxide and calcium carbonate.

[0148] Those skilled in the art will understand that specific counterions that form part of a pharmaceutically acceptable salt are usually not of critical nature, provided that the entire salt is pharmacologically acceptable and the counterion does not impart undesirable properties to the entire salt.

[0149] As described herein, the compounds of formulas (I) to (IV) may contain one or more free amino, hydroxy, carbonyl (e.g., keto or aldehyde) and / or carboxylic acid groups. In another case, the protected forms of the compounds of formulas (I) to (IV), in which the free amino, hydroxy, carbonyl (e.g., keto or aldehyde) and / or carboxylic acid groups are protected by appropriate protecting groups, are also encompassed by the present disclosure. The term "protecting group" refers to a chemical group that, when attached to a potentially reactive functional group, masks, reduces or prevents the reactivity of the functional group. Typically, the protecting group can be selectively removed when desired during the synthesis process.

[0150] Protecting groups are well-known in the art, and various examples are described, for example, in “Protective Groups in Organic Chemistry” (Greene, W. & Wuts, P. G. M., 2006, John Wiley & Sons). Examples of amino protecting groups include, but are not limited to, formyl, acetyl, trifluoroacetyl, benzyl (Bn), benzoyl (Bz), benzyloxycarbonyl (CBZ), tert-butoxycarbonyl (Boc), trimethylsilyl (TMS), 2-trimethylsilyl-ethanesulfonyl (TES), trityl, substituted trityl, tosyl, phthalimide, allyloxycarbonyl (Alloc), and 9-fluorenylmethyloxycarbonyl (FMOC). Examples of hydroxy protecting groups include, but are not limited to, acetyl, benzyl (Bn), t-butyl, benzoyl (Bz), β-methoxyethoxymethyl ether (MEM), dimethoxytrityl (DMT), methoxymethyl ether (MOM), methoxytrityl [(4-methoxyphenyl)diphenylmethyl] (MMT), p-methoxybenzyl ether (PMB), p-methoxyphenyl ether (PMP), methylthiomethyl ether, pivaloyl (Piv), tetrahydropyranyl (THP), tetrahydrofuran (THF), trityl, trimethylsilyl (TMS), tert-butyldimethylsilyl (TBDMS or TBS), triisopropylsilyloxymethyl (TOM), and tri-iso-propylsilyl (TIPS). Examples of carbonyl protecting groups include, but are not limited to, acetal, hemi-acetal, and ketal. Examples of carboxylic acid protecting groups include, but are not limited to, methyl ester, benzyl ester, tert-butyl ester, silyl ester, orthoester, and oxazoline.

[0151] Certain embodiments relate to pharmaceutically acceptable solvates of compounds having formulas (I)-(IV). Those skilled in the art will understand that certain compounds having formulas (I)-(IV) can combine with solvents such as water, methanol, ethanol, or acetonitrile to form pharmaceutically acceptable solvates such as the corresponding hydrates, methanolates, ethanolates or acetonitriles. Other examples of solvents that can be used to prepare solvates include isopropanol, dimethyl sulfoxide, ethyl acetate, acetic acid, ethanolamine and acetone, as well as miscible formulations of solvate mixtures that will be known to those skilled in the art.

[0152] In various embodiments, the compounds of the present disclosure, for example, compounds having a structure according to any one of formulas (I)-(IV), are agonists of TLR7. In some embodiments, the compounds of the present disclosure (e.g., compounds according to any one of formulas (I)-(IV)) can induce the production of one or more cytokines in immune cells (e.g., PBMC). In such embodiments, the compounds can induce the production of, for example, IL6, IFN-α and / or TNF-α in immune cells (e.g., PBMC) when such cells are contacted with the compounds.

[0153] In some embodiments, the compounds of the present disclosure, for example, compounds having a structure according to any one of formulas (I)-(IV), have an EC with respect to TLR7 agonism of about 1 μM or less, 750 nM or less, 650 nM or less, 500 nM or less, 300 nM or less, 275 nM or less, 250 nM or less, 225 nM or less, 200 nM or less, 175 nM or less, 150 nM or less, 125 nM or less, 100 nM or less, 75 nM or less, 50 nM or less, 25 nM or less, 20 nM or less, 15 nM or less, or about 10 nM or less as determined, for example, in a reporter gene assay 50may have. TLR7 may be human TLR7, mouse TLR7, or both. In some embodiments, any one compound of formulas (I)-(IV) has an EC 50 value of about 1 μM or less with respect to stimulating human and / or mouse TLR7. In some embodiments, any one compound of formulas (I)-(IV) has an EC 50 value of 500 nM or less with respect to stimulating human and / or mouse TLR7. In some embodiments, any one compound of formulas (I)-(IV) has an EC 50 value of 300 nM or less with respect to stimulating human and / or mouse TLR7. In some embodiments, any one compound of formulas (I)-(IV) has an EC 50 value of 100 nM or less with respect to stimulating human and / or mouse TLR7. In other embodiments, any one compound of formulas (I)-(IV) has an EC 50 value of 50 nM or less with respect to stimulating human and / or mouse TLR7. In yet another embodiment, any one compound of formulas (I)-(IV) has an EC 50 value of 25 nM or less with respect to stimulating human and / or mouse TLR7. As used herein, the EC 50 value generally refers to the half maximal effective concentration of each compound, and the value of EC 50 refers to the concentration of a compound that induces a baseline and maximal intermediate biological response (e.g., TLR7 agonism, stimulation of the immune system, cytokine production by immune cells, and / or killing of tumor cells) after a defined exposure period.

[0154] In some embodiments, the present disclosure relates to purine derivative compounds having an EC with respect to stimulating TLR7 of from about 1 μM to about 750 nM, from about 1 μM to about 500 nM, from about 1 μM to about 300 nM, from about 1 μM to about 200 nM, from about 1 μM to about 100 nM, from about 1 μM to about 50 nM, from about 500 nM to about 25 nM, from about 500 nM to about 15 nM, from about 500 nM to about 5 nM, from about 500 nM to about 1 nM, from about 500 nM to about 0.1 nM, from about 500 nM to about 0.01 nM, from about 300 nM to about 25 nM, from about 300 nM to about 15 nM, from about 300 nM to about 5 nM, from about 300 nM to about 1 nM, from about 300 nM to about 0.1 nM, from about 300 nM to about 0.01 nM, from about 200 nM to about 25 nM, from about 200 nM to about 15 nM, from about 200 nM to about 5 nM, from about 200 nM to about 1 nM, from about 200 nM to about 0.1 nM, from about 200 nM to about 0.01 nM, from about 100 nM to about 25 nM, from about 100 nM to about 15 nM, from about 100 nM to about 5 nM, from about 100 nM to about 1 nM, from about 100 nM to about 0.1 nM, from about 100 nM to about 0.01 nM, from about 50 nM to about 25 nM, from about 50 nM to about 15 nM, from about 50 nM to about 5 nM, from about 50 nM to about 1 nM, from about 50 nM to about 0.1 nM, or from about 50 nM to about 0.01 nM. 50 It relates to purine derivative compounds having the structure according to any one of formulas (I) to (IV), for example. In various embodiments, TLR7 is human TLR7.

[0155] The EC with respect to the TLR7 agonism of the compounds of the present disclosure, for example, compounds having the structure according to any one of formulas (I) to (IV). 50 Values can be determined using various methods known in the art, for example, using a reporter gene assay with TLR7 reporter cells in vitro. In such embodiments, the EC 50 Values can be determined using a reporter gene assay using HEK-Blue™ TLR7 reporter cells (available, for example, from Invivogen, San Diego, CA) in vitro, for example, as described in Example 3 herein.

[0156] In some embodiments, surprisingly, as further described herein, particularly when R 2 contains a substituent (i.e., R 2 ≠H), purine derivative compounds of formulas (I)-(IV) can provide potent immunomodulatory activity both in vitro and in vivo when used both as free molecules and in the form of immunopotentiating antibody-drug conjugates (ISACs). Such compounds can elicit their immunomodulatory activity in both mouse and human environments targeting mouse and human TLR7. In various embodiments, such a substituent at the R 2 position may be an alkoxy moiety or a halogen atom. In certain embodiments, the R 2 substituent may be methoxy or fluorine. Such compounds herein, when determined, for example, using a reporter gene assay (RGA) as described herein, may have an EC 50 of 500 nM or less, 100 nM, 50 nM, 30 nM, 20 nM, 10 nM or less, or lower with respect to stimulating TLR7.

[0157] In other embodiments, when R 2 is H, compounds of formula (I) in which (i) R 1 is a branched, optionally substituted C3-C8 hydroxyalkyl, (ii) X is NH and R 1 is an optionally substituted C5-C6 alkyl, or (iii) m is 0 or 1 and R 3 and R 4 together with N form an unsubstituted piperazinyl may have potent immunomodulatory properties that can be useful for the treatment of diseases such as cancer.

[0158] In some embodiments, the compounds of the present disclosure, such as compounds having a structure according to any one of formulas (I)-(IV), are each about 5 μM or less, 3 μM or less, 1 μM or less, 750 nM or less, about 500 nM or less, about 300 nM or less, about 275 nM or less, about 250 nM or less, about 225 nM or less, about 200 nM or less, about 175 nM or less, about 150 nM or less, about 125 nM, about 100 nM or less, about 50 nM or less, about 25 nM or less, or about 10 nM or less in terms of inducing the production of one or more human and / or mouse cytokines from human or mouse immune cells, with an EC 50 that may be present. In some embodiments, the compounds of the present disclosure, such as compounds having a structure according to any one of formulas (I)-(IV), are each about 5 μM to about 5 nM, about 1 μM to about 1 nM, about 750 nM to about 1 nM, about 500 nM to about 1 nM, about 300 nM to about 1 nM, or about 200 nM to about 1 nM in terms of inducing the production of one or more human and / or mouse cytokines from human or mouse immune cells, with an EC 50 that may be present. In various embodiments, the one or more human and / or mouse cytokines include IL6. In some embodiments, the human immune cells include PBMCs, and the mouse immune cells include mouse spleen cells. In various embodiments, the EC 50 values for inducing the production of human or mouse IL6 from PBMCs or mouse spleen cells are each determined using, for example, a human PBMC assay or a mouse spleen cell assay as described in Example 3 herein.

[0159] In some embodiments, the compounds of the present disclosure, such as compounds having a structure according to any one of formulas (I)-(IV), are about 10 nM or less, 5 nM or less, 1 nM or less, or 0.1 nM or less in terms of inducing the production of human interferon-α (hIFN-α), with an EC 50may have. In some embodiments, the compounds of the present disclosure, for example, compounds having a structure according to any one of formulas (I)-(IV), have an EC with respect to inducing the production of hIFN-α of from about 10 nM to about 1 nM, from about 1 nM to about 0.1 nM, or less than 0.1 nM 50 may have. In various embodiments, the EC with respect to inducing the production of hIFN-α from PBMC 50 values are determined using a human PBMC assay, as described, for example, in Example 3 herein.

[0160] In certain embodiments, the EC with respect to inducing the production of cytokines from PBMC by the compounds of the present disclosure, for example, compounds having a structure according to any one of formulas (I)-(IV) 50 is determined in vitro by treating PBMC isolated from peripheral blood with a concentration titration of the test compound, followed by assaying for the cytokine, for example, by homogeneous time-resolved fluorescence (HTRF). An exemplary method for determining the EC with respect to inducing the production of cytokines from PBMC 50 values is shown in Example 3 herein.

[0161] The purine derivative compounds of the present disclosure, for example, the compounds of formulas (I)-(IV), can be prepared from commercially available starting materials and reagents by standard synthetic organic chemical methods. Representative examples of suitable synthetic routes are described in detail in Example 1 shown herein (see also Figure 1). One of ordinary skill in the art will recognize that alternative methods are available for synthesizing the purine derivative compounds of the present disclosure, and thus, the approaches described herein are not intended to be exhaustive.

[0162] Compound-linker construct The purine derivative compounds of the present disclosure can be linked to a linker moiety. In certain embodiments, the present disclosure relates to a compound-linker construct comprising one or more compounds (C) of the present disclosure linked to a linker moiety (L), for example, a compound having a structure according to any one of formulas (I) to (IV), for example, any of the compounds listed in Table 1.

[0163] In various embodiments, the present disclosure provides a compound of formula (A): (A) L-(C) p relating to a compound-linker construct having (wherein, L is a linker; C is a compound of any one of formulas (I) to (IV); p is an integer from 1 to 5).

[0164] In various embodiments of the compound-linker construct, p may be an integer from 1 to 4. In other embodiments, p is an integer from 1 to 3. In yet another embodiment, p is an integer from 1 to 2. In various embodiments, p is 1. In other embodiments, p is 2. In yet another embodiment, p is 3.

[0165] The linker (L) herein may be a bifunctional or polyfunctional moiety capable of linking one or more immunomodulatory purine derivative compounds C of the present disclosure to another molecule, for example, a targeting moiety T, as further described elsewhere herein.

[0166] The bifunctional (or monovalent) linker L links one compound C to one site (e.g., a functional group) on the targeting moiety T. In such embodiments, p is 1. The polyfunctional (or polyvalent) linker L links two or more compounds C to one site (e.g., a functional group) on the targeting moiety T. A linker that links one compound C to two or more sites on the targeting moiety T can also be considered polyfunctional. However, in embodiments where the linker (L) is polyvalent and links two or more compounds (C) to another moiety, e.g., the targeting moiety (T), p can be an integer >1, e.g., from 2 to 5.

[0167] The linker L of the compound-linker constructs herein may include a functional group capable of reacting with a target group(s) on the targeting moiety T and at least one functional group capable of reacting with a target group on the compounds (e.g., compounds according to any one of formulas (I) - (IV)) C of the present disclosure. Suitable functional groups are known in the art and include, for example, those described in Bioconjugate Techniques (G.T. Hermanson, 2013, Academic Press). Targeting moieties T that can function as target groups for linker attachment and functional groups on the purine derivative compounds C of the present disclosure include, but are not limited to, thiol, hydroxyl, carboxyl, amine, aldehyde, and ketone groups.

[0168] In some embodiments, the linker herein comprises, or consists of, a substituted or unsubstituted alkyl or substituted or unsubstituted heteroalkyl chain containing first and second terminal functional groups. In some embodiments, the first terminal functional group of the linker forms a first linkage or bond with a first reactive functional group on a first conjugation partner (e.g., targeting moiety T), and the second terminal functional group of the linker forms a second linkage or bond with a second reactive functional group on a second conjugation partner (e.g., compound C of formulas (I)-(IV)). In some embodiments, the linker comprises a substituted or unsubstituted hydrocarbon backbone. In some embodiments, the substituted or unsubstituted hydrocarbon backbone is interrupted by one or more heteroatoms (e.g., O, N, S, P), thereby forming, for example, a heteroalkyl linker.

[0169] In some embodiments, the linker (L) is a cleavable linker. In other embodiments, the linker (L) is a non-cleavable linker. The cleavable linker herein is a linker that is readily cleaved under specific conditions, such as intracellular conditions (e.g., within endosomes or lysosomes) or in the vicinity of the target cell (e.g., tumor microenvironment). Examples include protease-sensitive, acid-sensitive, or reduction-sensitive linkers. In contrast, non-cleavable linkers may generally rely on the degradation of the targeting moiety (e.g., antibody) conjugated with the linker in the cell, which results in the release of the amino acid-linker-compound moiety. In some embodiments, examples of the linker include alkylene oxides, such as poly(alkylene oxides), or poly(ethylene glycol) (PEG) moieties.

[0170] In some embodiments, the linker (L) of the compound-linker construct herein comprises a poly(ethylene glycol) (PEG) moiety. Such a PEG moiety may have a molecular weight of from about 500 Da to about 5 kDa, from about 500 Da to about 3 kDa, from about 500 Da to about 1 kDa, from about 500 Da to about 1 kDa, or from about 100 Da to 500 Da.

[0171] Examples of cleavable linkers include, for example, linkers containing an amino acid sequence that is a cleavage recognition sequence of a protease. Many such cleavage recognition sequences are known in the art. In embodiments where the compound-linker constructs herein are used in complexes not intended to be internalized by cells, for example, amino acid sequences recognized and cleaved by proteases present in the extracellular matrix surrounding target cells such as cancer cells may be utilized. Examples of extracellular tumor-related proteases include, for example, plasmin, matrix metalloprotease (MMP), elastase, and kallikrein-related peptidases.

[0172] In embodiments where the compound-linker constructs herein are used in complexes intended to be internalized by cells, linker L may comprise an amino acid sequence recognized and cleaved by endosomal or lysosomal proteases. Examples of such proteases include, for example, cathepsin B, C, D, H, L, and S, as well as legumain.

[0173] The cleavage recognition array may be, for example, a dipeptide, tripeptide, or tetrapeptide. Non-limiting examples of dipeptide recognition arrays that may be included in the cleavable linkers described herein include Ala-(D)Asp, Ala-Lys, Ala-Phe, Asn-Lys, Asn-(D)Lys, Asp-Val, His-Val, Ile-Cit, Ile-Pro, Ile-Val, Leu-Cit, Me3Lys-Pro, Met-Lys, Met-(D)Lys, NorVal-(D)Asp, Phe-Arg, Phe-Cit, Phe-Lys, phenylGly-(D)Lys, Pro-(D)Lys, Trp-Cit, Val-Ala, Val-(D)Asp, Val-Cit, Val-Gly, Val-Gln, and Val-Lys, but are not limited thereto. Examples of tri- and tetrapeptide cleavage arrays include Ala-Ala-Asn, Ala-Val-Cit, (D)Ala-Phe-Lys, Asp-Val-Ala, Asp-Val-Cit, Gly-Cit-Val, Lys-Val-Ala, Lys-Val-Cit, Met-Cit-Val, (D)Phe-Phe-Lys, Asn-Pro-Val, Ala-Leu-Ala-Leu, Gly-Phe-Leu-Gly, Gly-Gly-Phe-Gly, and Gly-Phe-Gly-Gly, but are not limited thereto. In some embodiments, the linker (L) comprises a dipeptide, tripeptide, tetrapeptide, or a combination thereof. In some of these embodiments, the linker (L) comprises a dipeptide. In other embodiments, the linker (L) comprises a tripeptide. In yet other embodiments, the linker (L) comprises a tetrapeptide.

[0174] As further examples of cleavable linkers, there are included disulfide-containing linkers such as N-succinimidyl 4-(2-pyridyldithio) butanoate (SPDB) and N-succinimidyl 4-(2-pyridyldithio)-2-sulfobutanoate (sulfo-SPDB). Disulfide-containing linkers may optionally include additional groups that provide steric hindrance adjacent to the disulfide bond, such as, for example, geminal dimethyl groups, to improve the extracellular stability of the linker. Other cleavable linkers include linkers that are hydrolysable within a specific pH or pH range, such as hydrazone linkers. Linkers containing combinations of these functional groups may also be useful, for example, linkers containing both hydrazone and disulfide are known in the art.

[0175] A further example of a cleavable linker is a linker comprising a β-glucuronide that is cleavable by β-glucuronidase, an enzyme present in lysosomes and the tumor stroma (see, for example, De Graaf et al., 2002, Curr. Pharm. Des. 8: 1391-1403, and International Patent Publication No. WO2007 / 011968). β-Glucuronide can also function to improve the hydrophilicity of linker L.

[0176] Another example of a linker that is cleaved intracellularly and improves hydrophilicity is a linker that includes a pyrophosphate diester moiety (see, for example, Kern et al., 2016, J Am Chem Soc., 138: 2430-1445).

[0177] In certain embodiments, linker L included in the compound-linker construct of formula (A) is a cleavable linker. In some embodiments, such linker L includes a cleavage recognition sequence. In some embodiments, such linker L may include an amino acid sequence, for example, an amino acid sequence that is 2 or 3 residues in length and is recognized and cleaved by a protease.

[0178] Cleavable linkers may optionally further comprise one or more additional functional groups such as self-destructive groups and self-cleaving groups, stretchers, hydrophilic moieties, or combinations thereof.

[0179] Examples of self-destructive and self-cleaving groups that can be used for the linkers herein include, for example, p-aminobenzyl (PAB) and p-aminobenzyloxycarbonyl (PABC) groups, as well as methylated ethylenediamine (MED). Other examples of self-destructive groups include aromatic compounds electronically similar to PAB or PABC groups, such as heterocyclic derivatives, for example, 2-aminoimidazole-5-methanol derivatives described in U.S. Patent No. 7,375,078, but are not limited thereto. Other examples include groups that cyclize upon amide bond hydrolysis, such as substituted and unsubstituted 4-aminobutyramide (Rodrigues et al., 1995, Chemistry Biology 2:223-227) and 2-aminophenylpropionic acid amide (Amsberry, et al., 1990, J. Org. Chem. 55:5867-5877). Self-destructive / self-cleaving groups are generally attached to the amino or hydroxyl groups of compound C. Self-destructive / self-cleaving groups are often included in peptide-based linkers, either alone or in combination, but may also be included in other types of linkers.

[0180] Examples of stretchers that can be used for the linkers for the complexes further described herein include, for example, alkylene groups and fatty acid, dibasic acid, amine or diamine-based stretchers, such as diglycolate, malonate, caproate and caproamide. Other stretchers include, for example, glycine-based stretchers and polyethylene glycol (PEG) or monomethoxypolyethylene glycol (mPEG) stretchers.

[0181] PEG and mPEG stretchers can also function as hydrophilic moieties within the linkers of the present disclosure. For example, PEG or mPEG can be included in the linker either as an "inline" or pendant group to enhance the hydrophilicity of the linker (see, e.g., U.S. Patent Application Publication No. US2016 / 0310612). Various PEG-containing linkers are commercially available from companies such as Quanta BioDesign, Ltd (Plain City, OH). Other hydrophilic groups that may optionally be incorporated into linker L include, for example, β-glucuronide, sulfonate group, carboxylate group, and pyrophosphate diester.

[0182] In some embodiments, the compound-linker constructs of the present disclosure may comprise or consist of a compound of formula (I)-(IV) herein attached to any of the linker moieties L1-L18 shown in Table 2A (

Chemical formula

Table 2-1

Table 2-2

Table 2-3

[0183] In certain embodiments, the compound-linker construct of formula (A) may comprise a cleavable linker. In some embodiments, the compound-linker construct of formula (A) may comprise a peptide-containing linker. In some embodiments, the compound-linker construct of formula (A) may comprise a protease-cleavable linker.

[0184] In various embodiments, with respect to formula (I), the linker moiety (L) may be attached to a compound having the structure of formula (I) at any suitable atom. Suitable groups on compound C of formulas (I)-(IV) for attachment of linker L in any of the above approaches include, but are not limited to, thiol groups, amine groups, carboxylic acid groups, and hydroxyl groups. In some embodiments of the present disclosure, linker L is attached to the compound via a hydroxyl group or an amine group on compound C of formula (I).

[0185] In some embodiments, the attachment site of the linker moiety is one of the sites shown below by formula (I-L) based on formula (I), where "L" represents a potential linker attachment site.

Chemical formula

[0186] In some embodiments, the compound-linker constructs of the present specification containing any one of the purine derivative compounds of formulas (I)-(IV) may be selected from Table 2B.

Table 3-1

Table 3-2

Table 3-3

Table 3-4

Table 3-5

Table 3-6

Table 3-7

[0187] The compound-linker constructs of formula (A) in this specification can be prepared using any suitable synthetic technique known in the art, as further described herein, for example, in Example 2.

[0188] Complexes containing purine derivative compounds Complexes (also referred to herein as "immunopotentiating antibody-drug conjugates" or "ISACs") containing one or more disclosed compounds, for example, one or more compounds according to any one or more of formulas (I)-(IV), bound to a targeting moiety (T) are further disclosed herein. In various embodiments, the complexes of this specification are one or more compound-linker constructs of formula (A) described herein, L-(C) p comprise.

[0189] In some embodiments, the complexes of the present disclosure may comprise one or more compounds of any one of formulas (I)-(IV) conjugated to a targeting moiety (T). In various embodiments, the complexes of the present disclosure have the structure according to formula (X): (X) T-[L-(C) p r wherein, T is a targeting moiety, L is a linker; ​C is a compound according to any one of formulas (I) to (IV); p is an integer from 1 to 5; r has a value of from about 1 to about 8).

[0190] In various embodiments of the complex of formula (X) herein, L-(C) p is a compound-linker construct as described herein.

[0191] As further described herein, in certain embodiments, the compounds disclosed herein can be used for the preparation of a conjugate that is an immunopotentiating antibody-drug conjugate (ISAC) comprising an antibody as a targeting moiety that recognizes and binds to a tumor-associated antigen (e.g., a protein expressed on the surface of a targeted tumor cell). At the target site (e.g., a tumor), the disclosed immunopotentiating compound(s) conjugated to the targeting moiety is released within immune cells and / or within the tumor microenvironment (TME), for example, by using a cleavable linker, thereby resulting in an anti-tumor effect by activation of the immune system (e.g., by TLR7-related signaling and / or cytokine production).

[0192] A. Targeting moiety (T) The targeting moiety T included in the conjugate of formula (X) can be a molecule that binds to, reacts associatively with, or forms a complex with a receptor, antigen, or other receptor moiety associated with a given population of target cells. Generally, the targeting moiety T serves to deliver the purine derivative compound C of the present disclosure to a particular population of target cells with which the targeting moiety T reacts. Examples of targeting moieties include, but are not limited to, proteins (e.g., antibodies, antibody fragments, and growth factors), glycoproteins, peptides (e.g., bombesin and gastrin-releasing peptide), lectins, vitamins (e.g., folic acid), and nutrient transport molecules (e.g., transferrin).

[0193] Generally, the targeting moiety T is attached to the linker L via a heteroatom of the targeting moiety T such as sulfur (e.g., of a sulfhydryl group), oxygen (e.g., of a carbonyl, carboxyl or hydroxyl group) or nitrogen (e.g., of a primary or secondary amino group). These heteroatoms may originally be present in the targeting moiety T, or may be incorporated by manipulation and / or expression, or may be incorporated by chemical modification using techniques known in the art.

[0194] In some embodiments, the targeting moiety T is an antibody. Thus, since the compounds of the present disclosure can induce an immune response in a subject, various embodiments of the present disclosure relate to immunostimulatory antibody-drug conjugates (ISACs) having the general formula (X) where the targeting moiety T is an antibody.

[0195] In embodiments where the conjugate is an ISAC, the antibody included as the targeting moiety T may be a full-size polyclonal or monoclonal antibody, an antigen-binding antibody fragment (e.g., Fab, scFab, Fab’, F(ab’)2, Fv or scFv), a domain antibody (dAb) or an antibody mimetic (e.g., an affibody, DARPin, anticalin, versabody, duoCALIN, lipocalin or avimer). The antibody is generally directed against a specific antigen, e.g., a tumor-associated antigen (TAA).

[0196] In certain embodiments where the conjugate (X) is an ISAC, the targeting moiety T is a monoclonal antibody, an antigen-binding antibody fragment thereof (e.g., Fab, scFab, Fab’, F(ab’)2, Fv or scFv) or a domain antibody (dAb).

[0197] In certain embodiments, the targeting moiety T may be a monoclonal antibody. The monoclonal antibody may be, for example, a non-human monoclonal antibody (e.g., a mouse or rabbit antibody), a human monoclonal antibody, a humanized monoclonal antibody, or a chimeric antibody (e.g., a human-mouse antibody). In certain embodiments, the antibody included as the targeting moiety in the complex (X) herein may be a bispecific or multispecific antibody.

[0198] In certain embodiments, the targeting moiety T included in the complex is an antibody or an antigen-binding antibody fragment that binds to a tumor-associated antigen (TAA). In various embodiments, such a TAA-binding antibody may include a functional Fc region capable of binding to an Fc receptor.

[0199] B. Linker (L) used in the complex As further described herein, for example, in the section on "Compound-Linker Constructs", the linker can attach one or more of the immunomodulatory compounds of the present disclosure, such as any one of Compounds (I)-(IV), to the targeting moiety (T). For a detailed description of the linker moieties contemplated herein, see the section on "Compound-Linker Constructs" above.

[0200] As further described herein, the linker L of the compound-linker constructs herein may include a functional group capable of reacting with a target group(s) on the targeting moiety T, and at least one functional group capable of reacting with a target group (e.g., a compound C according to any one of Formulas (I)-(IV)) on the compounds of the present disclosure. Suitable functional groups are known in the art and include, for example, those described in Bioconjugate Techniques (G.T. Hermanson, 2013, Academic Press). Targeting moieties T that can function as target groups for linker attachment, and functional groups on the purine derivative compounds C of the present disclosure, include, but are not limited to, thiol, hydroxyl, carboxyl, amine, aldehyde, and ketone groups.

[0201] Non-limiting examples of functional groups capable of reacting with thiols include maleimide, haloacetamide, haloacetyl, activated esters (e.g., succinimide ester, 4-nitrophenyl ester, pentafluorophenyl ester, and tetrafluorophenyl ester), anhydrides, acid chlorides, sulfonyl chloride, isocyanate, and isothiocyanate. In this context, the "self-stabilizing" maleimide described in Lyon et al., 2014, Nat. Biotechnol., 32:1059-1062 is also useful.

[0202] Non-limiting examples of functional groups capable of reacting with amines include activated esters (e.g., N-hydroxysuccinamide (NHS) ester and sulfo-NHS ester), imido esters (e.g., Traut reagent), isothiocyanate, aldehyde, and acid anhydrides (e.g., diethylenetriaminepentaacetic anhydride (DTPA)). Other examples include succinimidyl-1,1,3,3-tetramethyluronium tetrafluoroborate (TSTU) and benzotriazol-1-yl-oxytripyrrolidinophosphonium hexafluorophosphate (PyBOP).

[0203] Non-limiting examples of functional groups capable of reacting with electrophilic groups such as aldehyde or ketone carbonyl groups include hydrazide, oxime, amino, hydrazine, thiosemicarbazone, hydrazine carboxylate, and aryl hydrazide.

[0204] In certain embodiments where the targeting moiety T is an antibody, the linker moiety L may comprise a functional group that enables cross-linking of two inter-chain cysteines on the antibody, such as a ThioBridge™ linker (Badescu et al., 2014, Bioconjug. Chem. 25:1124-1136), a dithiomaleimide (DTM) linker (Behrens et al., 2015, Mol. Pharm. 12:3986-3998), a dithioaryl (TCEP) pyridazinedione-based linker (Lee et al., 2016, Chem. Sci., 7:799-802) or a dibromopyridazinedione-based linker (Maruani et al., 2015, Nat. Commun., 6:6645).

[0205] In other embodiments, the targeting moiety T may be modified to include a non-natural reactive group, such as an azide, that enables conjugation to the linker via complementary reactive groups on the linker. For example, conjugation of the linker to the targeting moiety may utilize a click chemistry reaction, such as the azide-alkyne cycloaddition (AAC) reaction, which has been used successfully in the development of antibody-drug conjugates (see, e.g., Chio & Bane, 2020, Methods Mol. Biol., 2078:83-97). The AAC reaction may be a copper-catalyzed AAC (CuAAC) reaction involving the coupling of an azide with a linear alkyne, or a strain-promoted AAC (SPAAC) reaction involving the coupling of an azide with a cyclooctyne.

[0206] In some embodiments, the linker (e.g., cleavable) is substantially stable in the tumor extracellular environment, e.g., in circulation. Such a stable linker may be characterized in that at least about 90%, about 80%, about 70%, about 60%, about 50% or at least about 40% of the complex of formula X remains intact upon delivery to the tumor site and / or localization on the target cell surface and after a certain period of time. An intact complex may have an unchanged DAR (e.g., within about ±5%) compared to the complex at the time of administration, for example. In other words, in some embodiments, the linker remains substantially uncleaved in this environment while the complex is present in the tumor extracellular environment (e.g., in the systemic circulation and / or within non-target tissues or organs). In some embodiments, such a linker may be cleaved in the tumor extracellular environment, but not to the extent that it prevents delivery of an effective dose of the intact complex to the target cells (e.g., tumor cells). Whether the linker is substantially insensitive to the tumor extracellular environment can be determined, for example, by quantifying the amount of free compound (e.g., molecule cleaved from the complex) present in the plasma after incubating the ISAC with plasma for a predetermined period of time (e.g., 2, 4, 8, 16, or 24 hours).

[0207] Various cleavable linkers for linking the compounds of the present disclosure to the targeting moiety are known in the art and, in certain embodiments, may be used in the complexes of the present disclosure. Examples of cleavable linkers include N-succinimidyl ester or N-sulfosuccinimidyl ester moieties for reaction with cell binding agents, and linkers having maleimide or haloacetyl-based moieties for reaction with a compound (e.g., any one of the compounds according to formulas (I) to (IV)), or vice versa. Examples of such cleavable linkers are those based on sulfosuccinimidyl-4-[N-maleimidomethyl]cyclohexane-1-carboxylate (sulfo-SMCC). Sulfo-SMCC conjugation generally occurs via a maleimide group that reacts with the sulfhydryl (thiol, -SH) of compound C, while the sulfo-NHS ester is reactive towards the primary amines (found at the lysine and the N-terminus of proteins or peptides) of the targeting moiety T. Other non-limiting examples of such linkers include N-succinimidyl 4-(maleimidomethyl)cyclohexanecarboxylate (SMCC), N-succinimidyl-4-(N-maleimidomethyl)-cyclohexane-1-carboxy-(6-amidocaproate) (“long-chain” SMCC or LC-SMCC), N-succinimidyl maleimidoundecanoate (KMUA), N-succinimidyl maleimidobutyrate (GMBS), N-hydroxysuccinimide ester of maleimidocaproic acid (EMCS), m-maleimidobenzoyl-N-hydroxysuccinimide ester (MBS), N-(maleimidoacetoxy)-succinimide ester (AMAS), succinimidyl-6-(maleimidopropionamido)hexanoate (SMPH), 4-(p-maleimidophenyl)-butyric acid N-succinimidyl (SMPB) and those based on N-(p-maleimidophenyl)isocyanate (PMPI).Other examples include those containing haloacetyl-based functional groups such as N-succinimidyl-4-(iodoacetyl)-aminobenzoate (SIAB), N-succinimidyl iodoacetate (SIA), N-succinimidyl bromoacetate (SBA), and N-succinimidyl 3-(bromoacetamido)propionate (SBAP).

[0208] C. Preparation of the conjugate The conjugate of formula (X) herein can be prepared by standard methods known in the art (see, for example, Bioconjugate Techniques (G.T. Hermanson, 2013, Academic Press)). Various linkers and linker components are commercially available or can be prepared using standard synthetic organic chemistry techniques (see, for example, March’s Advanced Organic Chemistry (Smith & March, 2006, Sixth Ed., Wiley); Toki et al., (2002) J. Org. Chem. 67:1866-1872; Frisch et al., (1997) Bioconj. Chem. 7:180-186; Bioconjugate Techniques (G.T. Hermanson, 2013, Academic Press)). Additionally, various antibody-drug conjugation services from companies such as Lonza Inc. (Allendale, NJ), Abzena PLC (Cambridge, UK), ADC Biotechnology (St. Asaph, UK), Baxter BioPharma Solutions (Baxter Healthcare Corporation, Deerfield, IL), and Piramel Pharma Solutions (Grangemouth, UK) are commercially available.

[0209] Generally, the preparation of the conjugate described herein first involves preparing a compound-linker construct C-L of formula (A) described herein, which comprises one or more of the compounds of any one of formulas (I)-(IV) and a linker L, and then conjugating the compound-linker construct (C-L) p to a suitable group of the targeting moiety T. However, ligation of the targeting moiety-linker T-L to one or more of the compounds described herein, such as a compound C of formula (I), following ligation of the linker L and the targeting moiety T, is still an alternative approach available in some embodiments. Exemplary methods are described in Example 4 herein.

[0210] Suitable groups on the targeting moiety T for attachment of the linker L in any of the above approaches include sulfhydryl groups (e.g., in the side chain of a cysteine residue), amino groups (e.g., in the side chain of a lysine residue), carboxylic acid groups (e.g., in the side chain of an aspartic acid residue or a glutamic acid residue), and carbohydrate groups (e.g., in an Fc glycan moiety). For example, the targeting moiety T may include one or more naturally occurring sulfhydryl groups that allow the targeting moiety T to be attached to the linker L via the sulfur atom of the sulfhydryl group. In some embodiments, the targeting moiety T may include one or more lysine residues that are chemically modifiable to introduce one or more sulfhydryl groups. Reagents that can be used to modify lysine residues include, but are not limited to, N-succinimidyl S-acetylthioacetate (SATA), N-succinimidyl-3-(2-pyridyldithio)propionate (“SPDP”), and 2-iminothiolane hydrochloride (Traut reagent). In other embodiments, the targeting moiety T may include one or more carbohydrate groups that are chemically modifiable to include one or more sulfhydryl groups.

[0211] In some embodiments, the carbohydrate group on the targeting moiety T can also be oxidized to yield an aldehyde (-CHO) group (see, e.g., Laguzza et al., 1989, J. Med. Chem. 32(3):548 - 55), which can then be reacted with the linker L, for example, via a hydrazine group or a hydroxylamine group on the linker L.

[0212] The targeting moiety T can also be modified to contain additional cysteine residues (see, e.g., U.S. Patent Nos. 7,521,541; 8,455,622 and 9,000,130) or unnatural amino acids that provide reactive handles, such as selenomethionine, p - acetylphenylalanine, formylglycine, or p - azidomethyl - L - phenylalanine (see, e.g., Hofer et al., 2009, Biochemistry, 48:12047 - 12057; Axup et al., 2012, PNAS, 109:16101 - 16106; Wu et al., 2009, PNAS, 106:3000 - 3005; Zimmerman et al., 2014, Bioconj. Chem., 25:351 - 361) to enable site - specific conjugation. In some embodiments, the targeting moiety T may be modified to contain unnatural reactive groups, such as azide, that enable conjugation with the linker via complementary reactive groups on the linker, for example, by click chemistry (see, e.g., Chio & Bane, 2020, Methods Mol. Biol., 2078:83 - 97).

[0213] Other protocols for modifying proteins, which the targeting moiety (T) herein may include or consist of to enable binding or association of the linker L, are known in the art and include those described in Coligan et al., Current Protocols in Protein Science, vol. 2, John Wiley & Sons (2002).

[0214] In embodiments where the targeting moiety T is an antibody, several different reactive groups on the antibody, including the ε-amino group on lysine residues, pendant carbohydrate moieties, side chain carboxylic acid groups on aspartic acid or glutamic acid residues, cysteine-cysteine disulfide groups, and cysteine thiol groups, may function as conjugation sites. The amino acids used for conjugation may be part of the native sequence of the antibody or may be incorporated by site-specific manipulation techniques known in the art as described herein.

[0215] Once conjugation is complete, the average number of compounds of formulas (I)-(IV) conjugated to the targeting moiety T (i.e., the "drug-to-antibody ratio" or DAR) may be determined by standard techniques, such as UV / VIS spectroscopy, ELISA-based techniques, chromatographic techniques, such as hydrophobic interaction chromatography (HIC), UV-MALDI mass spectrometry (MS), and MALDI-TOF MS. Additionally, the distribution of the compound conjugation forms (e.g., the percentage of the targeting moiety T containing 0, 1, 2, 3, etc. compounds of formula (I)) may optionally be analyzed. A variety of techniques for evaluating the DAR distribution are known in the art, including MS (with or without an accompanying chromatographic separation step), hydrophobic interaction chromatography, reverse phase HPLC, or isoelectric focusing gel electrophoresis (IEF) (see, for example, Wakankar et al., 2011, mAbs, 3:161-172).

[0216] D. Characteristics of the Conjugate As described herein, the drug-to-antibody ratio (DAR) of the conjugate disclosed herein (or, in the case of r = 1, "p" of formula X, and in other cases the product of q × r) refers to the ratio of the number of compounds C of formulas (I)-(IV) conjugated to the targeting moiety T.

[0217] As noted above and as reflected by the parameters p and r of formula (X), the targeting moiety "T" can conjugate with two or more compounds "C" of formula (I) (plural possible). Those skilled in the art will understand that although any particular targeting moiety T is conjugated with an integer number of compound C, the analysis of the preparation of the conjugate for determining the ratio of compound C to targeting moiety T may result in a non-integer result that reflects a statistical average. This ratio of compound C to targeting moiety T is generally also called the drug-to-antibody ratio, or "DAR". Accordingly, conjugate preparations having a non-integer DAR are intended to be encompassed by formula (X). Those skilled in the art will understand that the term "DAR" may also be used to define conjugates that include targeting moieties other than antibodies.

[0218] In some embodiments, the DAR of the conjugate of formula (X) herein is obtained by any combination of linker-to-antibody ratio and / or drug-to-linker ratio (e.g., when r is the ratio of the compound-linker construct to the antibody T and p is the number of compound C per linker L1). For example, when r = 2 and p = 2, the number of compound C of formulas (I)-(IV) in each antibody-drug conjugate is 4 (e.g., each tumor-targeting antibody T is bound to two compound-linker constructs, and each linker construct contains two immunologically active compounds of formulas (I)-(IV)).

[0219] In some embodiments, the DAR of the conjugate of formula (X) is from about 1 to about 32. In some embodiments, the DAR of the conjugate of formula (X) is from about 1 to about 24, from about 1 to about 16, from about 1 to about 8, from about 3 to about 5, or from about 1 to about 4. In some embodiments, the DAR of the conjugate of formula (X) is from about 2 to about 32, from about 2 to about 24, from about 2 to about 16, from about 2 to about 8, or from about 2 to about 4. In various embodiments, the DAR may be from about 1.5 to about 4.5.

[0220] In some embodiments, the DAR of the conjugate of formula (X) herein (i.e., the product of r×p of formula (X)) may be any numerical value from about 1 to about 8, and thus may have a value of about 1.0, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, 2.0, 2.1, 2.2, 2.3, 2.4, 2.5, 2.6, 2.7, 2.8, 2.9, 3.0, 3.1, 3.2, 3.3, 3.4, 3.5, 3.6, 3.7, 3.8, 3.9, 4.0, 4.1, 4.2, 4.3, 4.4, 4.5, 4.6, 4.7, 4.8, 4.9, 5.0, 5.1, 5.2, 5.3, 5.4, 5.5, 5.6, 5.7, 5.8, 5.9, 6.0, 6.1, 6.2, 6.3, 6.4, 6.5, 6.6, 6.7, 6.8, 6.9, 7.0, 7.1, 7.2, 7.3, 7.4, 7.5, 7.6, 7.7, 7.8, 7.9, or about 8.0. In some embodiments, the DAR of the conjugate of formula (X) herein may have a value from about 2 to about 8, from about 2 to about 6, from about 2 to about 5, from about 3 to about 5, or from about 2 to about 4.

[0221] As further described elsewhere herein, all of the compounds of the conjugate of formula (X) (e.g., the compounds of formulas (I)-(IV)) may be the same. In other embodiments, two or more of the compounds of the conjugate of formula (X) may be different.

[0222] In some embodiments, the conjugate of formula (X) herein has an EC 50 value with respect to stimulating TLR7 of from about 10 nM to about 50 pM, from about 5 nM to about 50 pM, from about 1 nM to about 50 pM, from about 750 pM to about 50 pM, from about 600 pM to about 50 pM, from about 500 pM to about 50 pM, from about 400 pM to about 50 pM, from about 300 pM to about 50 pM, from about 200 pM to about 50 pM, or from about 100 pM to about 50 pM.

[0223] Although not bound by any theory, the antitumor activity of the complex of formula (X) herein is thought to potentially involve innate immune cells. The complex can first bind to tumor cells via the target TAA and then to immune cells via FcγR binding, particularly in embodiments where the targeting moiety is an antibody or a fragment thereof. The complex can then be degraded in the phagolysosome to release a TLR7 agonist, such as a compound of any one of formulas (I)-(IV). Stimulation of TLR7 in the phagolysosome can induce cytokine expression, which can drive an antitumor response. As a result, the TLR7 agonism involving the compounds described herein can drive antitumor immunity in a subject.

[0224] In various embodiments, as further described herein, the complex of formula (X) of the present disclosure comprising one or more immunomodulatory compounds can be capable of inducing the production and / or release of one or more cytokines from immune cells. Such immune cells can each be human or mouse immune cells such as PBMC or splenocytes, or can be part of a cell population further comprising other cell types such as other immune cells, tumor cells, etc. For example, such induction of inflammatory cytokines can be the result of an interaction between the immunostimulatory compound of the complex and TLR7 of the immune cells.

[0225] In various embodiments, the complex of formula (X) herein can induce the production of one or more cytokines when contacted with a cell population comprising immune cells. In various embodiments, such cytokines belong to the IL6 superfamily. In various embodiments, the complex of formula (X) herein can induce the production of IL6. The induction of IL6 can involve human IL6, mouse IL6, or both.

[0226] In various embodiments, the complex of formula (X) herein has an EC<1 nM, <500 pM, <250 pM, <100 pM, or <50 pM, or lower, with respect to inducing cytokine production in a human or mouse immune cell population. 50 In some embodiments, the complex of formula (X) herein has an EC<1 nM, <500 nM, or <250 nM with respect to inducing cytokine production in a human or mouse immune cell population. 50 In some embodiments, the complex of formula (X) herein has an EC of from about 1 nM to about 100 pM, from about 750 pM to about 50 pM, from about 500 pM to about 50 pM, from about 250 pM to about 50 pM, or from about 50 pM to about 50 pM with respect to inducing cytokine production in a human or mouse immune cell population. 50 In various embodiments, the cytokine whose production is induced by the complex is IL6. In some embodiments, the induction of cytokine production by the complex can be determined by a human and / or mouse immune cell assay, which may include, for example, incubating the complex with a cell population comprising immune cells (e.g., those expressing TLR7) and tumor cells (e.g., those expressing an antigen to which the targeting moiety binds), as further described herein in Example 5, followed by measuring the amount of cytokine in the supernatant after a certain period of time.

[0227] In various embodiments, a complex of formula (X) comprising a compound of any one of formulas (I)-(IV) herein can induce an immune response in vivo. In various embodiments, such an immune response can result in anti-tumor activity when administered to a subject having cancer (e.g., a human, non-human primate, or rodent). Such anti-tumor activity can be determined, for example, by a decrease in tumor growth rate. In some embodiments, when administered to a group of test subjects having tumors, the anti-tumor activity of the complex of formula (X) herein is sufficient to reduce the volume of the tumor in one or more test subjects. In some embodiments, the complex of formula (X) herein can reduce the volume of the tumor by at least about 30%, 40%, 50%, 60%, 70%, 80%, or 90% in at least one test subject.

[0228] In certain embodiments, the complex of formula (X) herein can exhibit good tolerance in vivo. In some embodiments, the complex of formula (X) herein can have good tolerance in vivo at a dose of at least about 1 mg / kg, at least about 3 mg / kg, at least about 15 mg / kg, or about 45 mg / kg. The complexes herein generally have good tolerance if the total body weight loss of the test subject at the end of the test period does not exceed 20% relative to the baseline weight at the start of the test, i.e., before treatment. The test period can be, for example, between 10 and 16 days. In some embodiments, the complex of formula (X) has good tolerance at a dose effective to reduce the volume of the tumor in the test subject.

[0229] Pharmaceutical composition The compounds of formulas (I)-(IV) and the complexes comprising the compounds of formulas (I)-(IV) may be formulated for therapeutic use. In certain embodiments, the present disclosure further discloses a pharmaceutical composition comprising a compound of the present disclosure, for example, a compound according to any one of formulas (I)-(IV) (see, for example, the section “Compounds” above), or a pharmaceutically acceptable salt thereof. A pharmaceutical composition comprising a complex of formula (X) is further disclosed herein (see, for example, the section “Complexes” above). Any such pharmaceutical composition may further comprise a pharmaceutically acceptable carrier, diluent, or additive. In some embodiments, the pharmaceutical composition is a therapeutic composition for the treatment thereof in a subject in need of treatment of a disease (e.g., cancer). Such pharmaceutical compositions can be prepared by known procedures using well-known and readily available ingredients.

[0230] The pharmaceutical compositions described herein may be formulated, for example, for administration to a subject by oral (e.g., buccal or sublingual), topical, parenteral, rectal, or vaginal routes, or by inhalation or insufflation. The term parenteral as used herein includes subcutaneous injection, and intradermal, intra-articular, intravenous, intramuscular, intracascular, intrasternal, intrathecal injection or infusion. The pharmaceutical compositions may be formulated in a manner suitable for administration to a subject, for example, as syrups, elixirs, tablets, troches, lozenges, hard capsules, soft capsules, pills, suppositories, oily suspensions, aqueous suspensions, dispersible powders, dispersible granules, emulsions, injections, or solutions. The pharmaceutical compositions may be provided as unit dosage formulations.

[0231] A pharmaceutical composition intended for oral use may be prepared in either solid or liquid unit dosage forms. Liquid unit dosage forms may be prepared according to procedures known in the art for the manufacture of pharmaceutical compositions, and such compositions may contain one or more agents such as sweetening agents, flavoring agents, coloring agents, and preservatives to provide a pharmaceutically elegant and palatable preparation. Elixirs can be prepared by using a hydroalcoholic (e.g., ethanol) carrier with an aromatic flavoring agent and a suitable sweetening agent such as sugar and / or saccharin. Suspensions can be prepared using an aqueous carrier and a suspending agent such as gum arabic, tragacanth, methylcellulose, etc.

[0232] Solid formulations such as tablets contain the active ingredient (e.g., the compounds and / or complexes of the present disclosure) in a mixture with non-toxic pharmaceutically acceptable additives suitable for the manufacture of tablets. These additives may be, for example, inert diluents such as calcium carbonate, sodium carbonate, lactose, calcium phosphate, or sodium phosphate; granulating and disintegrating agents such as corn starch or alginic acid; binders such as starch, gelatin, or gum arabic, and / or lubricants such as magnesium stearate, stearic acid, or talc, as well as other conventional ingredients such as calcium hydrogen phosphate, magnesium aluminum silicate, calcium sulfate, starch, lactose, methylcellulose, and functionally similar materials. Tablets may or may not be coated, or may be coated by known techniques, for example, to delay disintegration and absorption in the gastrointestinal tract, thereby providing a sustained action over a long period. For example, time-delay materials such as glyceryl monostearate or glyceryl distearate may be utilized.

[0233] Formulations for oral use may be presented as hard gelatin capsules in which the active ingredient (e.g., a compound and / or complex of the present disclosure) is mixed with an inert solid diluent such as calcium carbonate, calcium phosphate, or kaolin, or as soft gelatin capsules in which the active ingredient is mixed with an aqueous or oily medium such as peanut oil, liquid paraffin, or olive oil. Soft gelatin capsules are generally prepared by the mechanical encapsulation of a slurry of the active ingredient containing an acceptable vegetable oil, light liquid paraffin, or other inert oil.

[0234] An aqueous suspension may contain the active ingredient (e.g., a compound and / or complex of the present disclosure) in a mixture with additives suitable for the manufacture of an aqueous suspension. Such additives include suspending agents such as sodium carboxymethyl cellulose, methyl cellulose, hydroxypropyl methyl cellulose, sodium alginate, polyvinyl pyrrolidone, tragacanth gum, and acacia gum, and dispersing or wetting agents. Examples of dispersing and wetting agents include naturally occurring phospholipids (e.g., lecithin), condensates of alkylene oxide with fatty acids (e.g., polyoxyethylene stearate), condensates of ethylene oxide with long-chain aliphatic alcohols (e.g., heptadecaethyleneoxycetanol), condensates of ethylene oxide with partial esters derived from fatty acids and hexitol (e.g., polyoxyethylene sorbitol monooleate), or condensates of ethylene oxide with partial esters derived from fatty acids and hexitol anhydrides (e.g., polyethylene sorbitan monooleate). The aqueous suspension may also contain one or more preservatives (e.g., ethyl or n-propyl p-hydroxybenzoate), one or more coloring agents, one or more flavoring agents, and / or one or more sweetening agents (e.g., sucrose or saccharin).

[0235] The oily suspension can be formulated by suspending the active ingredient (e.g., the compounds and / or complexes of the present disclosure) in a vegetable oil such as peanut oil, olive oil, sesame oil, or coconut oil, or in a mineral oil such as liquid paraffin. The oily suspension may contain a thickening agent such as beeswax, solid paraffin, or cetyl alcohol. To provide a palatable oral preparation, sweeteners and flavoring agents such as those described above may be added. The suspension may optionally be protected by the addition of an antioxidant such as ascorbic acid.

[0236] Dispersible powders and granules suitable for the preparation of aqueous suspensions by the addition of water generally provide the active ingredient (e.g., the compounds and / or complexes of the present disclosure) in a mixture with a dispersing or wetting agent, a suspending agent, and one or more preservatives. Suitable dispersing or wetting agents, and suspending agents are exemplified by those already mentioned above. One or more additional additives such as sweeteners, flavoring agents, and / or coloring agents may also be present.

[0237] The pharmaceutical composition may also be in the form of a water-in-oil emulsion. The oil phase may be a vegetable oil such as olive oil or peanut oil, or a mineral oil such as liquid paraffin, or a mixture of such oils. Suitable emulsifying agents for inclusion in the water-in-oil emulsion include, for example, naturally occurring gums (such as acacia gum or tragacanth gum), naturally occurring phospholipids (such as soybean lecithin), or esters or partial esters derived from fatty acids and hexitol anhydrides (such as sorbitan monooleate) or condensates of such partial esters with ethylene oxide (such as polyoxyethylene sorbitan monooleate). The emulsion may also optionally contain sweeteners and / or flavoring agents.

[0238] The pharmaceutical compositions of the present disclosure may be in the form of sterile injectable aqueous or oily solutions or suspensions. Such suspensions can be formulated using suitable dispersing or wetting agents and suspending agents such as those mentioned above. The sterile injectable solution or suspension may contain the active ingredient (e.g., the compounds and / or complexes of the present disclosure) in a non-toxic parenterally acceptable carrier or diluent. Examples of available acceptable carriers and diluents include, for example, 1,3 - butanediol, water, Ringer's solution or isotonic sodium chloride solution. Furthermore, a sterile non - volatile oil may be used as a carrier. For this purpose, various non - irritating non - volatile oils containing synthetic mono - or diglycerides may be used. In addition, fatty acids such as oleic acid are also used in the preparation of injectables. Auxiliary agents such as local anesthetics, preservatives and / or buffering agents may also be included in the injectable solution or suspension.

[0239] The pharmaceutical compositions may also be formulated as suppositories for rectal administration. These compositions are solid at normal temperatures but liquid at physiological temperatures and can be prepared by mixing the active ingredient (e.g., the compounds and / or complexes of the present disclosure) with a suitable non - irritating additive that melts in the rectum and releases the drug. Examples of such materials include cocoa butter and polyethylene glycol.

[0240] Other pharmaceutical compositions and methods of preparing pharmaceutical compositions are known in the art and are described, for example, in “Remington: The Science and Practice of Pharmacy” (formerly “Remingtons Pharmaceutical Sciences”), Gennaro, A., Lippincott, Williams & Wilkins, Philadelphia, PA (2000).

[0241] Pharmaceutical kit In certain embodiments, a pharmaceutical composition comprising a compound of any one of formulas (I)-(IV) or a complex of formula (X) may be provided as part of a pharmaceutical kit or pack. The individual components of the kit may generally be packaged in separate containers. Suitable containers include, for example, bottles, blister packs, intravenous fluid bags, vials, etc., depending on the formulation of the pharmaceutical composition. In certain embodiments, the container may be in a form that enables administration to a subject, such as an inhaler, syringe, pipette, eye dropper, pre-moistened gauze or pad, or other device capable of administering the contents to the subject.

[0242] The kit may further include a label or package insert on or associated with the container(s). The term "package insert" is used to refer to the instructions customarily included in the commercial package of a therapeutic product, including information about the indications, usage, dosage, administration, contraindications, and / or warnings regarding the use of such therapeutic product. The label or package insert may further include a notice in a form determined by a government agency that regulates the manufacture, use or sale of the pharmaceutical or biological product, the notice indicating approval by the agency for manufacture, use or sale for administration to humans or animals. The label or package insert generally indicates the condition for which the compound or complex is selected, for example, that it is used for treating cancer.

[0243] If desired, one or more components of the kit may be lyophilized or provided in a dry form such as a powder or granule, and the kit may further include a solvent suitable for reconstituting the lyophilized or dried component(s).

[0244] Method of Use Certain embodiments of the present disclosure relate to the therapeutic use of purine-derived compounds described herein, such as compounds having a structure according to any one of formulas (I)-(IV) and complexes containing these compounds, such as complexes of formula (X). Some embodiments of the present disclosure relate to the use of a compound according to any one of formulas (I)-(IV) and / or a complex of formula (X) as a therapeutic agent.

[0245] The TLR7-stimulating compounds of the present disclosure (e.g., compounds according to any one of formulas (I)-(IV)) can exhibit cytotoxic activity against cancer cells. Therefore, a compound according to any one of formulas (I)-(IV) and a complex containing these compounds, i.e., a complex of formula (X), is useful for inhibiting the growth of abnormal cancer cells or tumor cells, for inhibiting cancer cell or tumor cell proliferation, and / or for treating cancer in a patient. In certain embodiments, a compound according to any one of formulas (I)-(IV) and a complex of formula (X) can be used to treat cancer in a subject in need of cancer treatment. Accordingly, some embodiments of the present disclosure relate to the use of a compound according to any one of formulas (I)-(IV) and a complex of general formula (X) as an anticancer agent.

[0246] Certain embodiments of the present disclosure relate to a method of inhibiting the growth of cancer or tumor cells, the method comprising contacting the cells with a compound according to any one of formulas (I)-(IV) or a complex of formula (X). Some embodiments relate to a method of killing cancer or tumor cells, the method comprising contacting the cells with a compound according to any one of formulas (I)-(IV) or a complex of formula (X).

[0247] Various embodiments of the present disclosure relate to methods of stimulating TLR7 using a compound of any one of formulas (I)-(IV) or a complex of formula (X) described herein. In some embodiments, a method of stimulating TLR7 in vitro, comprising contacting a cell expressing TLR7 with a compound of any one of formulas (I)-(IV) or a complex of formula (X) is disclosed herein. In other embodiments, a method of stimulating TLR7 in a subject, comprising administering to the subject a compound of any one of formulas (I)-(IV), a complex of formula (X), or a pharmaceutical composition comprising such a compound or complex is disclosed herein. Such methods may further comprise contacting a cell expressing TLR7 in the subject with a compound of any one of formulas (I)-(IV) or a complex of formula (X).

[0248] Stimulating TLR7 on cells in vitro or in vivo with the compounds or complexes of the present disclosure, such as a compound of any one of formulas (I)-(IV) or a complex of formula (X), can induce an immune response by the cells or the subject. In some embodiments, methods of inducing an immune response in vitro are disclosed herein, and such methods include contacting cells with a compound of any one of formulas (I)-(IV) or a complex of formula (X). The cells may be mammalian cells. The mammalian cells may be immune cells such as lymphocytes (e.g., T cells) or phagocytes (e.g., neutrophils, macrophages, dendritic cells, eosinophils, or monocytes). In yet another embodiment, a method of inducing an immune response in a subject in need thereof, the method comprising administering to the subject a compound of any one of formulas (I)-(IV) or a complex of formula (X) is disclosed herein. Such methods may further comprise stimulating TLR7 in the subject with the administered compound of any one of formulas (I)-(IV) or a complex of formula (X), thereby inducing an immune response in the subject. In various embodiments, methods for inducing a local immune response in a subject are disclosed herein, and such methods include administering to the subject in need thereof a complex of formula (X) herein comprising one or more of a compound of any one of formulas (I)-(IV). After administration of the complex of formula (X) to the subject (e.g., parenterally), such methods may further comprise accumulation of the complex at the target site (e.g., target organ or tissue) of the subject and inducing a local immune response at the target site. As used herein, the term "local immune response" generally refers to an immune response that occurs at a specific location within the subject, e.g., in an organ or within a specific tissue type present at one or more locations, in contrast to a systemic immune response. By way of example, a local anti-tumor immune response is generally understood herein to refer to an immune response that occurs at the tumor site, including the tumor tissue, or, in the case of metastases, at multiple tumor sites.Local immune responses can be determined or detected by changes in one or more physiological parameters such as local concentrations of biomarkers, such as production / secretion of cytokines, small molecule co-stimulatory molecules, and / or factors involved in inflammatory cascades or regulation, and / or changes in immune cell populations.

[0249] In some embodiments, methods of stimulating TLR7 using a compound of any one of formulas (I)-(IV) or a complex of formula (X) may further comprise activating the TLR7 signaling pathway. Such activation may cause a measurable change (i.e., an increase or decrease) in the expression of one or more intermediates in the TLR7 signaling pathway. Examples of intermediates in the TLR7 signaling pathway include, for example, MyD88, IRAK4, IRAK1, IRAK2, TRAF6, TAK1, IKK, NF-κB, FADD, caspase 8, caspase 3, and / or IRF7. See, e.g., Chi H et al. Front Pharmacol. 2017;8:304. Examples of inflammatory cytokines that can be modulated in response to incubation or contact with the compounds and / or complexes of the present disclosure include tumor necrosis factor (TNF; also known as TNFα or cachectin), interleukin (IL)-1α, IL1β, IL2; IL5, IL6, IL8, IL15, IL18, interferon γ (IFN-γ); platelet-activating factor (PAF), thromboxane; soluble adhesion molecules; vasoactive neuropeptides; phospholipase A2; plasminogen activator inhibitor (PAI-1); free radical generation; neopterin; CD14; prostacyclin; neutrophil elastase; protein kinase; monocyte chemoattractant protein 1 and 2 (MCP-1, MCP-2); macrophage migration inhibitory factor (MIF), and high-mobility group box protein 1 (HMGB-1), but are not limited thereto.

[0250] Methods for evaluating one or more of these physiological parameters are known in the art. For example, cytokines can be directly detected, for example, by ELISA. Other suitable methods include liquid chromatography and tandem mass spectrometry. Quantitative changes in biomolecules (e.g., cytokines) can be measured in biological samples such as organs, tissues, urine, or plasma. Detection of biomolecules may be performed directly on samples taken from a subject, or the samples may be processed between sample collection and analysis.

[0251] Some embodiments of the present disclosure relate to a method of treating a subject having cancer by administering to the subject a compound of formula (I) or a complex of formula (X), or a pharmaceutical composition comprising such a compound or complex. In this context, treatment with a compound of any one of formulas (I)-(IV) or a complex of formula (X) can result in one or more of: reduction in tumor size, delay or prevention of increase in tumor size, extension of the disease-free survival period between disappearance or removal of the tumor and recurrence, prevention of subsequent occurrence (e.g., metastasis) of the tumor, extension of the time to progression, reduction of one or more adverse symptoms associated with the tumor, and / or extension of the overall survival period of a subject having cancer.

[0252] Certain embodiments relate to the use of a compound of any one of formulas (I)-(IV) or a complex of formula (X) in a method of inhibiting tumor growth in a subject. Some embodiments relate to the use of a compound of any one of formulas (I)-(IV) or a complex of formula (X) in a method of inhibiting the growth of cancer cells and / or killing cancer cells in vitro. Some embodiments relate to the use of a compound of any one of formulas (I)-(IV) or a complex of formula (X) in a method of inhibiting the growth of cancer cells and / or killing cancer cells in vivo in a subject having cancer.

[0253] In certain embodiments of the disclosure using the compounds or complexes disclosed herein, examples of cancers treatable include hematological malignancies, including leukemia, myeloma, and lymphoma; carcinomas, including adenocarcinoma and squamous cell carcinoma; melanoma; and sarcoma. Carcinomas and sarcomas are also often referred to as "solid tumors." Examples of commonly occurring solid tumors that may be treated in certain embodiments include, but are not limited to, brain cancer, breast cancer, cervical cancer, colon cancer, head and neck cancer, kidney cancer, lung cancer, ovarian cancer, pancreatic cancer, prostate cancer, stomach cancer, uterine cancer, non-small cell lung cancer (NSCLC), and colorectal cancer. Various forms of lymphoma may also result in the formation of solid tumors and, thus, in certain situations may also be considered solid tumors.

[0254] In other embodiments, any one of the compounds of formulas (I)-(IV) herein, or the complex of formula (X), may be used as a vaccine adjuvant. In yet another embodiment, any one of the compounds of formulas (I)-(IV) herein, or the complex of formula (X), can be used in a method for treating a viral infection in a subject in need thereof. Such a method may include administering to the subject an effective amount of the compound or complex, thereby treating the viral infection in the subject.

[0255] Certain embodiments of the disclosure In certain embodiments, the disclosure relates to any one or more of embodiments 1-140.

[0256] Embodiment 1. Formula (I): [Chemical formula] The compound, its tautomer, protected form, and / or pharmaceutically acceptable salt (wherein, X is O or NH; Y is N or CH; R 1 is optionally substituted C2-C6 alkyl or branched optionally substituted C3-C8 hydroxyalkyl; R2 is H, halogen, optionally substituted C1-C6 alkyl, or optionally substituted C1-C6 alkoxy; R 3 and R 4 are independently H, Q-R 5 or R 3 and R 4 together with N form an optionally substituted C2-C6 heterocycloalkyl ring, an optionally substituted C3-C7 heterobicycloalkyl ring, an optionally substituted C6-C 10 -heterospirocycloalkyl ring or an optionally substituted heteroaryl ring; Q is a bond, optionally substituted C1-C6 alkyl, optionally substituted C2-C6 alkenyl or

Chemical formula

[0257] Embodiment 2. R 2 is halogen, optionally substituted C1-C6-alkyl, or optionally substituted C1-C6-alkoxy, a compound according to Embodiment 1.

[0258] Embodiment 3. R 2 is halogen or optionally substituted C1-C6-alkoxy, a compound according to any one of Embodiments 1-2.

[0259] Embodiment 4. R 2 is halogen or unsubstituted C1-C6-alkoxy, a compound according to any one of Embodiments 1-3.

[0260] Embodiment 5.R 2 The compound according to any one of Embodiments 1 to 4, wherein R is fluorine or methoxy.

[0261] Embodiment 6. The compound according to any one of Embodiments 1 to 5, wherein X is O.

[0262] Embodiment 7. The compound according to any one of Embodiments 1 to 5, wherein X is NH.

[0263] Embodiment 8.R 1 The compound according to any one of Embodiments 1 to 7, wherein R is optionally substituted C2-C6 alkyl.

[0264] Embodiment 9.R 1 The compound according to any one of Embodiments 1 to 7, wherein R is branched optionally substituted C3-C8 hydroxyalkyl.

[0265] Embodiment 10.R 3 and R 4 are independently H, or Q-R 5 The compound according to any one of Embodiments 1 to 9.

[0266] Embodiment 11.R 3 and R 4 are both H, or R 3 and R 4 are both Q-R 5 The compound according to any one of Embodiments 1 to 10.

[0267] Embodiment 12.R 3 is H, and R 4 is Q-R 5 The compound according to any one of Embodiments 1 to 10.

[0268] Embodiment 13. Q is a bond, and R 5is an optionally substituted C3-C6-carboxyalkyl, optionally substituted C3-C6-cycloalkyl, optionally substituted C2-C6-heterocycloalkyl, optionally substituted C3-C7-heterobicycloalkyl, optionally substituted C6-C 10 -heterospirocycloalkyl, optionally substituted aryl, or optionally substituted heteroaryl, the compound according to Embodiment Error! Reference source not found..

[0269] Embodiment 14. Q is an optionally substituted C1-C6-alkyl, and R 5 is H, OH, NR 6 R 7 , optionally substituted C3-C6-carboxyalkyl, optionally substituted C3-C6-cycloalkyl, optionally substituted C2-C6-heterocycloalkyl, optionally substituted C3-C7-heterobicycloalkyl, optionally substituted C6-C 10 -heterospirocycloalkyl, optionally substituted aryl, or optionally substituted heteroaryl, the compound according to Embodiment Error! Reference source not found..

[0270] Embodiment 15. Q is an optionally substituted C2-C6-alkenyl, and R 5 is H, OH, NR 6 R 7 , optionally substituted C3-C6-carboxyalkyl, optionally substituted C3-C6-cycloalkyl, optionally substituted C2-C6-heterocycloalkyl, optionally substituted C3-C7-heterobicycloalkyl, optionally substituted C6-C 10 -heterospirocycloalkyl, optionally substituted aryl, or optionally substituted heteroaryl, the compound according to Embodiment Error! Reference source not found..

[0271] Embodiment 16. Q is

Chemical formula

[0272] Embodiment 17. Q is

Chemical formula

[0273] Embodiment 18. R 5 is NR 6 R 7 wherein R 6 and R 7 are both H, a compound according to any one of Embodiments 1 to! Reference source not found! or! Reference source not found! to! Reference source not found!.

[0274] Embodiment 19. R 5 is NR 6 R 7 wherein R 6 is H and R 7 is optionally substituted C1-C6 alkyl, a compound according to any one of Embodiments 1 to! Reference source not found! or! Reference source not found! to! Reference source not found!.

[0275] Embodiment 20. R 5 is NR 6 R 7 wherein R 6 and R 7is a C2-C6 heterocycloalkyl ring optionally substituted together with N, an optionally substituted C3-C7 heterobicycloalkyl ring, an optionally substituted C6-C 10 -heterospirocycloalkyl ring or an optionally substituted heteroaryl ring, a compound according to any one of Embodiments 1 to Error! Reference source not found. Or Error! Reference source not found.~Error! Reference source not found.

[0276] Embodiment 21.R 3 and R 4 is a C2-C6 heterocycloalkyl ring optionally substituted together with N, an optionally substituted C3-C7 heterobicycloalkyl ring, an optionally substituted C6-C 10 -heterospirocycloalkyl ring or an optionally substituted heteroaryl ring, a compound according to any one of Embodiments 1 to 9.

[0277] Embodiment 22. n is 1, a compound according to any one of Embodiments 1 to 21.

[0278] Embodiment 23. m is an integer from 0 to 3 or from 1 to 3, a compound according to any one of Embodiments 1 to 22.

[0279] Embodiment 24.R 2 is H, and R 1 is a branched optionally substituted C3-C8 hydroxyalkyl, a compound according to Embodiment 1.

[0280] Embodiment 25.R 1 is a branched unsubstituted C3-C8 hydroxyalkyl, a compound according to Embodiment 13.

[0281] Embodiment 26.R 2 is H, X is NH, and R 1 is an optionally substituted C5-C6 alkyl, a compound according to Embodiment 1.

[0282] Embodiment 27.R 1The compound according to the embodiment, which is unsubstituted C5-C6 alkyl. Error! Reference source not found.

[0283] Embodiment 28. R 2 is H, m is 0 or 1, and R 3 and R 4 together with N form an unsubstituted piperazinyl ring or an optionally substituted C6-C 10 -heterospirocycloalkyl ring. The compound according to Embodiment 1.

[0284] Embodiment 29. R 3 and R 4 together with N form an unsubstituted piperazinyl ring. The compound according to the embodiment, Error! Reference source not found.

[0285] Embodiment 30. R 3 and R 4 together with N form an optionally substituted C6-C 10 -heterospirocycloalkyl ring. The compound according to the embodiment, Error! Reference source not found.

[0286] Embodiment 31. m is 0. The compound according to any one of the embodiments, Error! Reference source not found. - Error! Reference source not found.

[0287] Embodiment 32. m is 1. The compound according to any one of the embodiments, Error! Reference source not found. - Error! Reference source not found.

[0288] Embodiment 33. Each alkyl, alkenyl, cycloalkyl, spirocycloalkyl, heterocycloalkyl, heterospirocycloalkyl, aryl, and heteroaryl group is optionally substituted with -NH2, -CO2H, -OH, carbonyl, halogen, C1-C4 alkyl, C1-C4 hydroxyalkyl, C1-C4 aminoalkyl, C1-C4 carboxyalkyl, (C0-C4 alkyl)-cycloalkyl, (C0-C4 alkyl)-heterocycloalkyl, (C0-C4 alkyl)-spirocycloalkyl, (C0-C4 alkyl)-heterospirocycloalkyl, (C0-C4 alkyl)-aryl, and (C0-C4 alkyl)-heteroaryl, and each alkyl, hydroxy, C1-C4 aminoalkyl, C1-C4 carboxyalkyl, cycloalkyl, heterocycloalkyl, spirocycloalkyl, heterospirocycloalkyl, aryl, or heteroaryl group itself may be substituted with one or more of -NH2, -CO2H, -OH, carbonyl, halogen, unsubstituted C1-C4 alkyl, unsubstituted C1-C4 hydroxyalkyl, unsubstituted C1-C4 aminoalkyl, or unsubstituted C1-C4 carboxyalkyl, a compound according to any one of Embodiments 1 to Error! Reference source not found..

[0289] Embodiment 34. Each alkyl, alkenyl, cycloalkyl, spirocycloalkyl, heterocycloalkyl, heterospirocycloalkyl, aryl, and heteroaryl group is optionally substituted with one or more substituents selected from the group consisting of -NH2, -CO2H, -OH, carbonyl, halogen, unsubstituted C1-C4 alkyl, unsubstituted C1-C4 hydroxyalkyl, unsubstituted C1-C4 aminoalkyl, and unsubstituted C1-C4 carboxyalkyl, a compound according to Embodiment Error! Reference source not found..

[0290] Embodiment 35. The compound is selected from any one of Compounds 100 to 195 listed in Table 1, a compound according to Embodiment 1.

[0291] Embodiment 36. Formula (II): [Chemical formula] The compound according to Embodiment 1 having the structure of (wherein, X is O or NH; Y is N or CH; R 1 is optionally substituted C2-C6 alkyl or branched optionally substituted C3-C8 hydroxyalkyl; R 3 and R 4 are independently H, Q-R 5 or R 3 and R 4 together with N form an optionally substituted C2-C6 heterocycloalkyl ring, an optionally substituted C3-C7 heterobicycloalkyl ring, an optionally substituted C6-C 10 -heterospirocycloalkyl ring or an optionally substituted heteroaryl ring; Q is a bond, optionally substituted C1-C6 alkyl, optionally substituted C2-C6 alkenyl or [Chemical formula] wherein, * is the bonding point with R 5 and # is the bonding point with N; R 5 is H, OH, NR 6 R 7 optionally substituted C3-C6 carboxyalkyl, optionally substituted C3-C6 cycloalkyl, optionally substituted C2-C6 heterocycloalkyl, optionally substituted C3-C7 heterobicycloalkyl, optionally substituted C6-C 10 -heterospirocycloalkyl, optionally substituted aryl, or optionally substituted heteroaryl; R 6 and R 7 are independently H, optionally substituted C1-C6 alkyl, optionally substituted C1-C6 alkoxycarbonyl or R 6 and R 7together with N forms an optionally substituted C2-C6-heterocycloalkyl ring, an optionally substituted C3-C7-heterobicycloalkyl ring, an optionally substituted C6-C 10 -heterospirocycloalkyl ring or an optionally substituted heteroaryl ring; W is an optionally substituted C3-C6-cycloalkyl, an optionally substituted C2-C6-heterocycloalkyl, an optionally substituted C3-C7-heterobicycloalkyl, an optionally substituted C6-C 10 -heterospirocycloalkyl, an optionally substituted aryl, or an optionally substituted heteroaryl; m is an integer from 0 to 4; (wherein a and b are independently integers from 0 to 4).

[0292] Embodiment 37. The compound according to embodiment 15, wherein X is O and R 1 is an optionally substituted C2-C6 alkyl.

[0293] Embodiment 38. The compound according to any one of embodiments 15 to!Reference source not found!, wherein R 1 is unsubstituted C2-C6 alkyl.

[0294] Embodiment 39. The compound according to any one of embodiments 15 to!Reference source not found!, wherein Y is CH.

[0295] Embodiment 40. The compound according to any one of embodiments 15 to!Reference source not found!, wherein m is 0 or 1, 1 or 2, or 1 or 3.

[0296] Embodiment 41. The compound according to any one of embodiments 15 to!Reference source not found!, wherein R 3 and R 4 are both H.

[0297] Embodiment 42. The compound according to any one of embodiments 15 to!Reference source not found!, wherein R 3 and R 4is a C2-C6 heterocycloalkyl ring optionally substituted together with N, an optionally substituted C3-C7 heterobicycloalkyl ring, an optionally substituted C6-C 10 -heterospirocycloalkyl ring or an optionally substituted heteroaryl ring, a compound according to any one of embodiments 15 to error! Reference source not found.

[0298] Embodiment 43.R 3 and R 4 are both H or Q-R 5 either, or R 3 is H and R 4 is Q-R 5 a compound according to any one of embodiments 15 to error! Reference source not found.

[0299] Embodiment 44. Q is a bond and R 5 is an optionally substituted C3-C6 carboxyalkyl, an optionally substituted C3-C6 cycloalkyl, an optionally substituted C2-C6 heterocycloalkyl, an optionally substituted C3-C7 heterobicycloalkyl, an optionally substituted C6-C 10 -heterospirocycloalkyl, an optionally substituted aryl, or an optionally substituted heteroaryl, a compound according to embodiment error! Reference source not found.

[0300] Embodiment 45. Q is an optionally substituted C1-C6 alkyl and R 5 is H, OH, NR 6 R 7 , an optionally substituted C3-C6 carboxyalkyl, an optionally substituted C3-C6 cycloalkyl, an optionally substituted C2-C6 heterocycloalkyl, an optionally substituted C3-C7 heterobicycloalkyl, an optionally substituted C6-C 10 -heterospirocycloalkyl, an optionally substituted aryl, or an optionally substituted heteroaryl, a compound according to embodiment error! Reference source not found.

[0301] Embodiment 46. Q is optionally substituted C2-C6-alkenyl, and R 5 is H, OH, NR 6 R 7 , optionally substituted C3-C6-carboxyalkyl, optionally substituted C3-C6-cycloalkyl, optionally substituted C2-C6-heterocycloalkyl, optionally substituted C3-C7-heterobicycloalkyl, optionally substituted C6-C 10 -heterospirocycloalkyl, optionally substituted aryl, or optionally substituted heteroaryl, the compound according to Embodiment Error! Reference source not found.

[0302] Embodiment 47. R 5 is NR 6 R 7 , and R 6 and R 7 are both H, the compound according to any one of Embodiments 15-Error! Reference source not found. or Error! Reference source not found.~Error! Reference source not found.

[0303] Embodiment 48. R 5 is NR 6 R 7 , and R 6 is H, and R 7 is optionally substituted C1-C6 alkyl, the compound according to any one of Embodiments 15-Error! Reference source not found. or Error! Reference source not found.~Error! Reference source not found.

[0304] Embodiment 49. R 6 and R 7 together with N form an optionally substituted C2-C6-heterocycloalkyl ring, an optionally substituted C3-C7-heterobicycloalkyl ring, an optionally substituted C6-C 10 -heterospirocycloalkyl ring or an optionally substituted heteroaryl ring, the compound according to any one of Embodiments 15-Error! Reference source not found. or Error! Reference source not found.~Error! Reference source not found.

[0305] Embodiment 50.m is a compound according to any one of Embodiments 15 to !Reference source not found! which is 1 or 2, or 1 or 3.

[0306] Embodiment 51. The compound is a compound according to Embodiment 36 selected from Sub-Table 1A.

[0307] Embodiment 52.R 3 and R 4 together with N

Chemical formula

[0308] Embodiment 53.R 3 and R 4 together with N

Chemical formula

[0309] Embodiment 54. The compound is a compound according to any one of Embodiments 1 to 9 or 21 selected from Sub-Table 1B.

[0310] Embodiment 55.R 3 and R 4 both are Q-R 5The compound according to any one of Embodiments 1 to Error! Reference source not found..

[0311] Embodiment 56. The compound is

Chemical formula

[0312] Embodiment 57. R 3 and R 4 are both H, the compound according to any one of Embodiments 1 to 11.

[0313] Embodiment 58. The compound is

Chemical formula

[0314] Embodiment 59. The compound is selected from Sub-table 1C, the compound according to Embodiment 13.

[0315] Embodiment 60. The compound is

Chemical formula

[0316] Embodiment 61. The compound is selected from Sub-table 1D, the compound according to any one of Embodiments 1 to 10 or 14.

[0317] Embodiment 62. R 6 is H, R 7 is H, optionally substituted C1-C6 alkyl, or R 6 and R 7 together with N are an optionally substituted C2-C6 heterocycloalkyl ring, an optionally substituted C3-C7 heterobicycloalkyl ring, an optionally substituted C6-C 10-A compound according to any one of Embodiments 1 to 10 or 16 to 17, which forms a heterospirocycloalkyl ring or an optionally substituted heteroaryl ring.

[0318] Embodiment 63. The compound is a compound according to Embodiment 62, which is selected from Sub-Table 1E.

[0319] Embodiment 64. The compound is

Chemical formula

[0320] Embodiment 65. The compound is

Chemical formula

[0321] Embodiment 66. Formula (III):

Chemical formula

Chemical formula

[0322] Embodiment 67.R 1a is the compound according to Embodiment 28, which is an optionally substituted C2-C6 alkyl.

[0323] Embodiment 68.R 1a is the compound according to any one of Embodiments 28 to Error! Reference source not found., which is an unsubstituted C2-C6 alkyl.

[0324] Embodiment 69.R 2a is the compound according to any one of Embodiments 28 to Error! Reference source not found., which is a halogen or an optionally substituted C1-C6 alkoxy.

[0325] Embodiment 70.R 2a is the compound according to any one of Embodiments 28 to Error! Reference source not found., which is a halogen or an unsubstituted C1-C6 alkoxy.

[0326] Embodiment 71.R 2a is the compound according to any one of Embodiments 28 to Error! Reference source not found., which is fluorine or methoxy.

[0327] Embodiment 72.R 3a and R 4a together form an optionally substituted C2-C6 heterocycloalkyl ring, an optionally substituted C3-C7 heterobicycloalkyl ring, an optionally substituted C6-C 10 heterospirocycloalkyl ring or an optionally substituted heteroaryl ring, which is the compound according to any one of Embodiments 28 to Error! Reference source not found.

[0328] Embodiment 73.R 3a and R 4a are both H, which is the compound according to any one of Embodiments 28 to Error! Reference source not found.

[0329] Embodiment 74.R 3a and R 4ais, independently, H, or Q-R 5 A compound according to any one of Embodiments 28 to Error! Reference source not found.

[0330] Embodiment 75.R 3a is H, and R 4a is Q-R 5a A compound according to the embodiment of Error! Reference source not found.

[0331] Embodiment 76.Q is a bond, and R 5a is optionally substituted C3-C6-carboxyalkyl, optionally substituted C3-C6-cycloalkyl, optionally substituted C2-C6-heterocycloalkyl, optionally substituted C3-C7-heterobicycloalkyl, optionally substituted C6-C 10 -heterospirocycloalkyl, optionally substituted aryl, or optionally substituted heteroaryl, a compound according to any one of Embodiments Error! Reference source not found. to Error! Reference source not found.

[0332] Embodiment 77.Q is optionally substituted C1-C6-alkyl, and R 5a is H, OH, NR 6a R 7a , optionally substituted C3-C6-carboxyalkyl, optionally substituted C3-C6-cycloalkyl, optionally substituted C2-C6-heterocycloalkyl, optionally substituted C3-C7-heterobicycloalkyl, optionally substituted C6-C 10 -heterospirocycloalkyl, optionally substituted aryl, or optionally substituted heteroaryl, a compound according to any one of Embodiments Error! Reference source not found. to Error! Reference source not found.

[0333] Embodiment 78.Q is optionally substituted C2-C6-alkenyl, and R 5a is H, OH, NR 6a R 7a, optionally substituted C3-C6-carboxyalkyl, optionally substituted C3-C6-cycloalkyl, optionally substituted C2-C6-heterocycloalkyl, optionally substituted C3-C7-heterobicycloalkyl, optionally substituted C6-C 10 -heterospirocycloalkyl, optionally substituted aryl, or optionally substituted heteroaryl, a compound according to any one of embodiments error! Reference source not found.~error! Reference source not found.

[0334] Embodiment 79. Q is [Chemical formula] wherein W is optionally substituted C3-C6-cycloalkyl, optionally substituted C2-C6-heterocycloalkyl, optionally substituted C3-C7-heterobicycloalkyl, optionally substituted C6-C 10 -heterospirocycloalkyl, optionally substituted aryl, or optionally substituted heteroaryl; a and b are independently integers from 0 to 4, a compound according to any one of embodiments 28~error! Reference source not found.

[0335] Embodiment 80. Q is [Chemical formula] selected from the group consisting of, wherein a and b are independently 1, 2, or 3, a compound according to embodiment error! Reference source not found.

[0336] Embodiment 81. R 6a and R 7a are both H, a compound according to any one of embodiments 28~error! Reference source not found. or error! Reference source not found.~error! Reference source not found.

[0337] Embodiment 82. R 6a is H, R7a is an optionally substituted C1-C6 alkyl, a compound according to any one of Embodiments 28 to! Reference source not found. Or! Reference source not found.~! Reference source not found.

[0338] Embodiment 83.R 6a and R 7a is an optionally substituted C2-C6 heterocycloalkyl ring, an optionally substituted C3-C7 heterobicycloalkyl ring, an optionally substituted C6-C 10 -heterospirocycloalkyl ring or an optionally substituted heteroaryl ring, a compound according to any one of Embodiments 28 to! Reference source not found. Or! Reference source not found.~! Reference source not found.

[0339] In Embodiment 84, u is 1, a compound according to any one of Embodiments 28 to! Reference source not found.

[0340] In Embodiment 85, v is an integer from 0 to 4, from 1 to 4, from 1 to 3, or from 1 to 2, a compound according to any one of Embodiments 28 to! Reference source not found.

[0341] Embodiment 86.R 3a and R 4a together with N

Chemical formula

[0342] Embodiment 87.R 3a and R 4a together with N

Chemical formula

[0343] Embodiment 88. Formula (IV):

Chemical formula

Chemical formula

[0344] Embodiment 89. Each alkyl, alkenyl, cycloalkyl, spirocycloalkyl, heterocycloalkyl, heterospirocycloalkyl, aryl and heteroaryl group is optionally substituted with -NH2, -CO2H, -OH, carbonyl, halogen, C1-C4 alkyl, C1-C4 hydroxyalkyl, C1-C4 aminoalkyl, C1-C4 carboxyalkyl, (C0-C4 alkyl)-cycloalkyl, (C0-C4 alkyl)-heterocycloalkyl, (C0-C4 alkyl)-spirocycloalkyl, (C0-C4 alkyl)-heterospirocycloalkyl, (C0-C4 alkyl)-aryl, and (C0-C4 alkyl)-heteroaryl, and each alkyl, hydroxy, C1-C4 aminoalkyl, C1-C4 carboxyalkyl, cycloalkyl, heterocycloalkyl, spirocycloalkyl, heterospirocycloalkyl, aryl or heteroaryl group itself may be substituted with one or more of -NH2, -CO2H, -OH, carbonyl, halogen, unsubstituted C1-C4 alkyl, unsubstituted C1-C4 hydroxyalkyl, unsubstituted C1-C4 aminoalkyl, or unsubstituted C1-C4 carboxyalkyl, a compound according to Embodiment 66 or Embodiment 88.

[0345] Embodiment 90. Each alkyl, alkenyl, cycloalkyl, spirocycloalkyl, heterocycloalkyl, heterospirocycloalkyl, aryl and heteroaryl group is optionally substituted with one or more substituents selected from the group consisting of -NH2, -CO2H, -OH, carbonyl, halogen, unsubstituted C1-C4 alkyl, unsubstituted C1-C4 hydroxyalkyl, unsubstituted C1-C4 aminoalkyl and unsubstituted C1-C4 carboxyalkyl, a compound according to Embodiment 30.

[0346] Embodiment 91. The compound is selected from Sub-Table 1A, a compound according to Embodiment 66 or Embodiment 88.

[0347] Embodiment 92. The compound is

Chemical formula

[0348] Embodiment 93. The compound has an EC value for stimulating human or mouse TLR7 of <500 nM, <250 nM, or <100 nM as determined in a reporter gene assay, and / or an EC value for inducing IL6 production in a human or mouse immune cell population of <5 μM, <1 μM, <500 nM, or <100 nM as determined in an immune cell assay, and is the compound according to any one of Embodiments 1 to 92. 50 value, and / or an EC value for inducing IL6 production in a human or mouse immune cell population of <5 μM, <1 μM, <500 nM, or <100 nM as determined in an immune cell assay. 50 The compound according to any one of Embodiments 1 to 92, having the above value.

[0349] Embodiment 94. A pharmaceutical composition comprising the compound according to any one of Embodiments 1 to 92 and a pharmaceutically acceptable carrier or diluent.

[0350] Embodiment 95. A method for stimulating TLR7, comprising contacting a cell expressing TLR7 with the compound according to any one of Embodiments 1 to 33, thereby stimulating TLR7.

[0351] Embodiment 96. A method for inducing the release of cytokines from cells, comprising contacting the cells with a compound according to any one of Embodiments 1 to 33, thereby inducing the release of the cytokines from the cells.

[0352] Embodiment 97. A method for stimulating an immune response in a subject in need thereof, comprising administering to the subject an effective amount of the compound according to any one of Embodiments 1 to 33.

[0353] Embodiment 98. The method according to Embodiment Error! Reference source not found., wherein the compound stimulates TLR7 in the subject, thereby stimulating the immune response in the subject.

[0354] Embodiment 99. A method for inhibiting the growth of cancer cells, the method comprising contacting a cell population containing the cancer cells with an effective amount of the compound according to any one of Embodiments 1 to 33.

[0355] Embodiment 100. A method for killing cancer cells, the method comprising contacting a cell population containing the cancer cells with an effective amount of the compound according to any one of Embodiments 1 to 33.

[0356] Embodiment 101. The method according to any one of Embodiments error! Reference source not found.~error! Reference source not found., wherein the cell population contains immune cells.

[0357] Embodiment 102. A method for treating cancer in a subject in need of treatment for cancer, the method comprising administering to the subject an effective amount of the compound according to any one of Embodiments 1 to 33.

[0358] Embodiment 103. The method according to Embodiment error! Reference source not found., wherein the compound stimulates TLR7 in the subject, thereby treating the cancer in the subject.

[0359] Embodiment 104. The compound according to any one of Embodiments 1 to 92 for use in therapy.

[0360] Embodiment 105. The compound according to any one of Embodiments 1 to 92 for use in the treatment of cancer.

[0361] Embodiment 106. Use of the compound according to any one of Embodiments 1 to 92 in the manufacture of a medicament for the treatment of cancer.

[0362] Embodiment 107. Formula (A): (A) L-(C) p A compound-linker construct having (wherein, L is a linker; C is a compound according to any one of Embodiments 1 to 92; p is an integer from 1 to 5).

[0363] Embodiment 108. The compound-linker construct according to Embodiment 107, wherein L is a cleavable linker.

[0364] Embodiment 109. The compound-linker construct according to Embodiment 107, wherein L is a non-cleavable linker.

[0365] Embodiment 110. The compound-linker construct according to any one of Embodiments 107 to 108, wherein L comprises a dipeptide, a tripeptide, a tetrapeptide, or a combination thereof.

[0366] Embodiment 111. The compound-linker construct according to any one of Embodiments 107 to 110, wherein L comprises a polyethylene glycol (PEG) moiety.

[0367] Embodiment 112. The compound-linker construct according to any one of Embodiments 107 to 108, wherein L is a protease-cleavable linker.

[0368] Embodiment 113. The compound-linker construct according to any one of Embodiments 107 to 112, wherein C is a compound according to Embodiment 36.

[0369] Embodiment 114. The compound-linker construct according to any one of Embodiments 107 to 112, wherein C is a compound according to Embodiment 66.

[0370] Embodiment 115. The compound-linker construct according to Embodiment 107, wherein the compound-linker is selected from Table 2B.

[0371] Embodiment 116. The compound-linker construct according to any one of Embodiments 107 to 115, wherein p is 1, 2, or 3.

[0372] Embodiment 117. Formula (X): (X) T - [L - (C) p r having a complex (wherein, T is a targeting moiety, L is a linker; C is a compound according to any one of Embodiments 1 to 33; p is an integer from 1 to 5; r has a value from about 1 to about 8).

[0373] Embodiment 118. L - (C) p is a compound - linker construct according to any one of Embodiments 107 to 116, the complex according to Embodiment 117.

[0374] Embodiment 119. r is 1.0, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, 2.0, 2.1, 2.2, 2.3, 2.4, 2.5, 2.6, 2.7, 2.8, 2.9, 3.0, 3.1, 3.2, 3.3, 3.4, 3.5, 3.6, 3.7, 3.8, 3.9, 4.0, 4.1, 4.2, 4.3, 4.4, 4.5, 4.6, 4.7, 4.8, 4.9, 5.0, 5.1, 5.2, 5.3, 5.4, 5.5, 5.6, 5.7, 5.8, 5.9, 6.0, 6.1, 6.2, 6.3, 6.4, 6.5, 6.6, 6.7, 6.8, 6.9, 7.0, 7.1, 7.2, 7.3, 7.4, 7.5, 7.6, 7.7, 7.8, 7.9, or 8.0, the complex according to any one of Embodiments 117 to 118.

[0375] Embodiment 120. C is a compound according to Embodiment 36, the complex according to Embodiment 117.

[0376] Embodiment 121. C is a compound according to Embodiment 66 or Embodiment 88, the complex according to Embodiment 117.

[0377] Embodiment 122. C is selected from Sub - Table 1F, the complex according to Embodiment 117. ​

[0378] Embodiment 123.T is the conjugate according to any one of Embodiments 117 to 122 that binds to a tumor-associated antigen (TAA).

[0379] Embodiment 124.T is the conjugate according to any one of Embodiments 117 to 123, which is an antibody or an antigen-binding antibody fragment.

[0380] Embodiment 125.The conjugate according to Embodiment 124, wherein the antibody is a bispecific or multispecific antibody.

[0381] Embodiment 126.The conjugate according to any one of Embodiments 124 to 125, wherein the antibody or the antigen-binding antibody fragment binds to a TAA.

[0382] Embodiment 127.The conjugate according to any one of Embodiments 117 to 126 has an EC 50 value for inducing cytokine production in a human or mouse immune cell population of <1 nM, <500 pM, or <100 pM as determined in a reporter gene assay.

[0383] Embodiment 128.A pharmaceutical composition comprising the conjugate according to any one of Embodiments 117 to 126 and a pharmaceutically acceptable carrier or diluent.

[0384] Embodiment 129.A method of stimulating TLR7, the method comprising contacting a cell expressing TLR7 with the conjugate according to any one of Embodiments 117 to 126, thereby stimulating TLR7.

[0385] Embodiment 130.A method of inducing cytokine release from a cell, the method comprising contacting the cell with a conjugate according to any one of Embodiments 117 to 126, thereby inducing the cytokine release from the cell.

[0386] Embodiment 131. A method for stimulating an immune response in a subject in need of stimulation of the immune response, the method comprising administering to the subject an effective amount of the complex according to any one of Embodiments 117 to 126.

[0387] Embodiment 132. The method according to Embodiment 132, wherein the complex stimulates TLR7 in the subject, thereby stimulating the immune response in the subject.

[0388] Embodiment 133. A method for inhibiting the growth of cancer cells, the method comprising contacting a cell population containing the cancer cells with an effective amount of the complex according to any one of Embodiments 117 to 126.

[0389] Embodiment 134. A method for killing cancer cells, the method comprising contacting a cell population containing the cancer cells with an effective amount of the complex according to any one of Embodiments 117 to 126.

[0390] Embodiment 135. The method according to any one of Embodiments 133 to 134, wherein the cell population contains immune cells.

[0391] Embodiment 136. A method for treating cancer in a subject in need of treatment of cancer, the method comprising administering to the subject an effective amount of the complex according to any one of Embodiments 117 to 126.

[0392] Embodiment 137. The method according to Embodiment 136, wherein the complex stimulates TLR7 in the subject, thereby treating the cancer in the subject.

[0393] Embodiment 138. The complex according to any one of Embodiments 117 to 126 for use in therapy.

[0394] Embodiment 139. The complex according to any one of Embodiments 117 to 126 for use in the treatment of cancer.

[0395] Use of the complex according to any one of Embodiments 117 to 126 in the manufacture of a medicament for the treatment of cancer.

Example

[0396] The following examples are provided for illustration and are not intended to limit the scope of the present invention in any way.

[0397] The following examples provide exemplary methods for generating and using the compounds of the present disclosure, for example, any one compound of Formulas (I) to (IV). Of course, those skilled in the art can synthesize these compounds by similar methods or by combining other methods known in the art. The preparation of other compounds of Formulas (I) to (IV) not explicitly shown herein will be achieved by those skilled in the art using the methods described herein or similar methods with appropriate starting components and, if necessary, changes in the synthesis parameters. Generally, the starting components and materials can be obtained from commercial suppliers such as Sigma Aldrich (Merck KGaA), Alfa Aesar and Maybridge (Thermo Fisher Scientific Inc.), Matrix Scientific, Tokyo Chemical Industry Ltd. (TCI), and Fluorochem Ltd., and / or can be synthesized according to materials known to those skilled in the art (see, for example, March’s Advanced Organic Chemistry: Reactions, Mechanisms, and Structure, 7th edition, John Wiley & Sons, Inc., 2013), or can be prepared as described herein.

[0398] The following abbreviations are used throughout the examples: DCM = dichloromethane; DIPEA = N,N - diisopropylethylamine; DMA = dimethylacetamide; DMF = dimethylformamide; DMSO = dimethyl sulfoxide; IL6 = interleukin 6; LC / MS = liquid chromatography / mass spectrometry; LC / MSD = liquid chromatography / mass selective detector; SEC = size exclusion chromatography; HIC = hydrophobic interaction chromatography; RP - UPLC = reversed - phase ultra - performance liquid chromatography; HPLC = high - performance liquid chromatography; MT = maleimidotriethylene glycolate; PABC = p - aminobenzyl - oxycarbonyl; PBMC = peripheral blood mononuclear cell; PNP = p - nitrophenol; rt = room temperature; TCEP = tris(2 - carboxyethyl)phosphine; TFA = trifluoroacetic acid; TNF - α = tumor necrosis factor α; VC = valine - citrulline; UHPLC = ultra - high - performance liquid chromatography.

[0399] General Chemical Procedures General Procedure 1: Conversion from Chloride to Amine To a chloride compound in DMF (50 - 100 mg / mL), a primary or secondary amine (2 - 5 equivalents) was added, followed by DIPEA (3 - 5 equivalents), and the solution was heated to 50 - 75 °C with stirring. Upon completion (generally 1 - 18 hours), the reaction mixture was adjusted to pH 2 with 6M HCl and purified by reverse - phase HPLC to obtain the desired product after lyophilization.

[0400] General Procedure 2: Removal of Boc Protecting Group TFA (10 volume %) was added to a stirred solution (0.1M) of a Boc - protected amine compound in dichloromethane. Upon completion (generally 1 hour), the reaction mixture was concentrated in vacuo and co - evaporated with DCM several times to obtain a crude solid or purified by preparative HPLC to obtain the desired product after lyophilization.

[0401] General Procedure 3: Formation of Benzyl Chloride The alcohol compound was dissolved in 10% SOCl2 / DCM (0.05 - 0.1 M). The solution was stirred at room temperature until completion (generally 12 - 24 hours), and then concentrated in vacuo. The residue was co-evaporated (2 x 10 mL of dichloromethane) to give the crude chloride compound, which was generally used without further purification.

[0402] General Procedure 4: Synthesis of MT-VC-PABC Compound-Linker Constructs To a primary or secondary amine compound in dimethylformamide (0.05 - 0.1 M), the compound of Example 2.5 (MT-VC-PABC-PNP) (1 - 1.1 equivalents) was added, followed by DIPEA (3 equivalents). At completion (generally 1 - 4 hours), the reaction mixture was acidified with aqueous HCl (1 M) and then purified by reverse phase HPLC to give the desired drug-linker after lyophilization.

[0403] General Procedure 5: Purification of Compounds Flash Chromatography: The crude reaction product was purified using a Biotage® Snap Ultra column (10, 25, 50, or 100 g) (Biotage, Charlotte, NC) on a Biotage® Isolera™ automated flash system (Biotage, Charlotte, NC) eluting with a linear gradient of ethyl acetate / hexane or methanol / dichloromethane. Alternatively, a Biotage® Snap Ultra C18 column (12, 30, 60, or 120 g) was used and eluted with a linear gradient of CH3CN + 0.1% TFA / H2O + 0.1% TFA for reverse phase flash purification. The purified compound was isolated either by removal of the organic solvent by rotary evaporator or by lyophilization of the acetonitrile / water mixture.

[0404] Preparative HPLC: Reverse-phase HPLC of the crude compound was carried out on an Agilent 1260 Infinity II Preparative LC-MSD system (Agilent Technologies, Inc., Santa Clara, CA) using a Kinetex® 5-μm EVO C18 100Å (250×21.2 mm) column (Phenomenex), eluting with a linear gradient of CH3CN + 0.1% TFA / H2O + 0.1% TFA. The purified compound was isolated by lyophilization of the acetonitrile / water mixture.

[0405] General Procedure 6: Analysis of Compounds LC-MS: The completion of the reaction was monitored, and the purified compound was analyzed on an Agilent 1290 HPLC / 6120 single quad LC-MS system (Agilent Technologies, Inc., Santa Clara, CA) using a Kinetex® 2.6-μm C18 100Å (30×3 mm) column (Phenomenex), eluting with a linear gradient of 10 to 100% CH3CN + 0.1% TFA / H2O + 0.1% TFA.

[0406] NMR: 1 1H NMR spectra were collected using a Bruker AVANCE III 300 Spectrometer (300 MHz) (Bruker Corporation, Billerica, MA). Chemical shifts are in parts per million (ppm).

[0407] General Procedure 7: Synthesis of MT-VK-PABC Compound-Linker Constructs To a primary or secondary amine compound in dimethylformamide (0.05 - 0.1 M), the compound of Example 2.38 ((MT-VK(Boc)-PABC-PNP) (1.0 - 1.1 equivalents) was added, followed by DIPEA (3 - 5 equivalents). HOBt (1.0 equivalent) was used in some reactions as indicated. Upon completion (generally 1 - 4 hours), the reaction mixture was acidified with aqueous HCl (1 M) and then purified by reverse phase HPLC to obtain the Boc-protected drug-linker after lyophilization. The Boc intermediate was then deprotected according to General Procedure 2. Upon completion (generally 1 hour), the reaction mixture was concentrated in vacuo and co-evaporated several times with DCM to obtain a crude solid or purified by preparative HPLC to obtain the desired product after lyophilization.

[0408] General Procedure 8: Conversion from mesylate to amine To a chloride compound in DMF (50 - 100 mg / mL), a primary or secondary amine (2 - 5 equivalents) and NaI (0.1 - 1.0 equivalent) were added, followed by DIPEA (3 - 5 equivalents), and the solution was heated to 80 °C with stirring. Upon completion (generally 1 - 18 hours), the reaction mixture was adjusted to pH 2 with 6 M HCl and purified by reverse phase HPLC to obtain the desired product after lyophilization.

[0409] General Procedure 9: Preparation of ISAC using maleimide-containing compound-linker constructs Antibodies (1 - 10 mg / mL in phosphate - buffered saline, pH 7.4) were reduced with TCEP (1 - 10 mM in dH2O) (1.0 - 3.0 equivalents) in the presence of 1 mM DTPA. The solution was mixed well and incubated at 37 °C for 120 minutes, then cooled on ice. Subsequently, the reduced antibody solution was further buffer - exchanged into 10 mM sodium acetate buffer, pH 4.5, by passing it through a Zeba™ Spin Desalting column (40KDa MWCO; Thermo Scientific™). To the reduced protein solution stored on ice, a maleimide - functionalized compound - linker construct (10 mM in DMSO) (12 - 20 equivalents) was added. In some embodiments, propylene glycol (10 - 30 percent v / v%) was added to the reduced protein solution prior to the addition of the compound - linker construct. The conjugation reaction was immediately mixed well by pipetting and the conjugation was allowed to proceed at room temperature for 120 to 180 minutes. Upon completion, the reaction mixture was purified by passing it through a Zeba™ Spin Desalting column (40KDa MWCO; Pierce) pre - equilibrated with 10 mM sodium acetate, pH 4.5. The purified conjugate was stored at 4 °C and analyzed for total protein content characterized by HPLC - HIC, SEC, and / or RP - UPLC - MS (bicinchoninic acid assay, Pierce micro - BCA protocol, catalog number 23225). The average DAR and drug distribution were derived from the interpretation of HIC and / or LC - MS data. The average DAR estimate of the synthesized conjugate was in the range of about 1.5 to 5, or more specifically 1.9 to 4.3, as further shown herein. Endotoxin levels were evaluated at a sensitivity of 0.005 EU / mL in an Endosafe® nexgen - PTS™ test system using an Endosafe® LAL test cartridge (Charles River catalog number PTS20005F). Residual free compound and compound - linker construct levels were evaluated by RP - UPLC - MS with a threshold set at 1% ((free compound + compound - linker construct) / (conjugated compound - linker construct)).

[0410] Example 1: Preparation of Compounds of Formulas (I) - (IV) Example 1.1: 2,6 - Dichloro - 9 - (tetrahydro - 2H - pyran - 2 - yl) - 9H - purine

Chemical Structure

[0411] Example 1.2: 2 - Chloro - 9 - (tetrahydro - 2H - pyran - 2 - yl) - 9H - purin - 6 - amine

Chemical Structure

[0412] Example 1.3: 9 - Benzyl - 2 - chloro - 9H - purin - 6 - amine

Chemical Structure

[0413] Example 1.4: 1-((tert-butyldiphenylsilyl)oxy)pentan-2-ol

Chem.

[0414] Example 1.5: 2-Ethoxy-9-(tetrahydro-2H-pyran-2-yl)-9H-purin-6-amine [Chemical formula] To a solution of compound 2 (20.0 g, 78.8 mmol, 1.00 equiv) in EtOH (250 mL) was added KOtBu (17.7 g, 157 mmol, 2.0 equiv). The resulting solution was stirred at 60 °C for 18 h, then the solvent was removed in vacuo. The resulting solid was suspended in H2O (150 mL) and extracted with EtOAc (3 × 250 mL). The aqueous layer was further extracted with 10% iPrOH / EtOAc (100 mL). The pooled organics were washed with brine (50 mL), dried over MgSO4, and concentrated in vacuo to give the title compound 5 as an orange solid (20.0 g, 76.0 mmol, 96.4%). LC-MS: C 12 H 17 Calculated m / z for C11H14N5O2 = 263.1, found [M+H] + = 264.2.

[0415] Example 1.6: 8-Bromo-2-ethoxy-9-(tetrahydro-2H-pyran-2-yl)-9H-purin-6-amine [Chemical formula] To a suspension of compound 5 (20.0 g, 76.0 mmol, 1.00 equiv) in DCM (200 mL) was added NBS (20.3 g, 91.2 mmol, 1.20 equiv) portionwise over 3 min. The resulting mixture was stirred at room temperature for 18 h, then the reaction was quenched by the addition of 1 M NaHSO3 (50 mL) and stirred rapidly for 30 min. The resulting mixture was diluted with DCM (100 mL), and the separated organic layer was washed with 1 M NaHSO3 (2 × 50 mL). The pooled organics were dried over MgSO4, filtered, and concentrated in vacuo to give the title compound 6 as an orange solid (23.5 g, 68.7 mmol, 90.4%). LC-MS: C 12 H 16 Calculated m / z for C11H13BrN5O2 = 341.1, found [M+H] += 342.2。

[0416] Example 1.7: 2-Ethoxy-8-methoxy-9-(tetrahydro-2H-pyran-2-yl)-9H-purin-6-amine

Chem.

[0417] Example 1.8: 2-Ethoxy-8-methoxy-9-(tetrahydro-2H-pyran-2-yl)-9H-purin-6-amine

Chem.

[0418] Example 1.9: Methyl 4-((6-amino-2-ethoxy-8-methoxy-9H-purin-9-yl)methyl)-3-methoxy-benzoate [Chem.] To a solution of compound 8 (0.70 g, 3.4 mmol, 1.0 equiv) and methyl 4-(bromomethyl)-3-methoxybenzoate (0.87 g, 3.4 mmol, 1.0 equiv) in DMF (8 mL) was added CsCO3 (1.1 g, 3.4 mmol, 1.0 equiv). The resulting suspension was stirred at room temperature for 18 h and then concentrated in vacuo. Reverse-phase flash purification was carried out using a 60 g C18 column eluting with a gradient of 10 to 60% CH3CN + 0.1% TFA / H2O + 0.1% TFA as described in General Procedure 5. The title compound 9 was obtained as a white solid (0.65 g, 1.7 mmol, 50%). LC-MS: C 18 H 21 Calculated m / z = 387.2 for C18H19N5O5, found [M+H] + = 388.2.

[0419] Example 1.10: (4-((6-Amino-2-ethoxy-8-methoxy-9H-purin-9-yl)methyl)-3-methoxyphenyl)methanol [Chem.] To a stirred solution of compound 9 (250 mg, 0.65 mmol, 1.0 equiv) in anhydrous THF (5 mL) cooled to 0 °C was added lithium aluminum hydride (24 mg, 0.65 mmol, 1.0 equiv) portionwise over 5 min. The resulting suspension was warmed to room temperature and stirred for 15 min, then quenched with H2O (1 mL), diluted with MeOH (100 mL), and then filtered through a plug of celite. The filtrate was concentrated in vacuo, redissolved in 50% CH3CN / H2O (25 mL), and freeze-dried to give the title compound 10 (230 mg, 0.63 mmol, 97%). LC-MS: C 17 H 21 Calculated m / z = 359.2 for C17H19N5O4, found [M+H] + = 360.2.

[0420] Example 1.11: 6-Amino-9-(4-(chloromethyl)-2-methoxybenzyl)-2-ethoxy-9H-purin-8-ol

Chem.

[0421] ClN5O3, found [M+H]

Chem.

[0422] Example 1.13: 6-Amino-2-ethoxy-9-(4-(((2-hydroxy-2-methylpropyl)amino)methyl)-2-methoxybenzyl)-9H-purin-8-ol (Compound 101)

Chemical formula

[0423] Example 1.14: 6-Amino-9-(4-((diethylamino)methyl)-2-methoxybenzyl)-2-ethoxy-9H-purin-8-ol (Compound 102) [Chemical formula] The title compound was prepared from Compound 11 (0.010 g, 0.028 mmol, 1.0 equivalent) and diethylamine (7.0 mg, 0.082 mmol, 3.0 equivalents) according to General Procedure 1. Preparative HPLC purification was carried out with a gradient of 10 to 45% CH3CN + 0.1% TFA / H2O + 0.1% TFA for elution as described in General Procedure 5, and the title compound 102 was obtained as a white solid (10 mg, 0.026 mmol, 95%). LC-MS: C 20 H 28 Calculated m / z = 400.2 for C19H26N6O3, found [M+H] + = 401.2. 1 H NMR (300 MHz, CD3CN) δ 7.13 - 7.05 (m, 2H), 6.97 (dd, J = 7.7, 1.6 Hz, 1H), 4.97 (s, 2H), 4.31 (q, J = 7.0 Hz, 2H), 4.19 (s, 2H), 3.87 (s, 3H), 3.11 (septet, J = 7.2 Hz, 4H), 1.30 - 1.23 (m, 9H).

[0424] Example 1.15: 6-Amino-2-ethoxy-9-(4-(((5-hydroxypentyl)amino)methyl)-2-methoxybenzyl)-9H-purin-8-ol (Compound 103) [Chemical formula] The title compound was prepared from Compound 11 (0.010 g, 0.028 mmol, 1.0 equiv) and 5-aminopentan-1-ol (9.0 mg, 0.03 mmol, 3 equiv) according to General Procedure 1. Preparative HPLC purification was carried out with a gradient of 10 to 45% CH3CN + 0.1% TFA / H2O + 0.1% TFA as described in General Procedure 5 to obtain the title compound 103 as a white solid (10 mg, 0.015 mmol, 95%). LC-MS: C 21 H 30 Calculated m / z for C + H 1 N6O4 = 430.2, found [M+H]

[0425] Example 1.16: 6-Amino-9-(4-(((3-(4-(3-aminopropyl)piperazin-1-yl)propyl)amino)methyl)-2-methoxybenzyl)-2-ethoxy-9H-purin-8-ol (Compound 104)

Chemical Structure

[0426] Example 1.17: 6-Amino-9-(4-(((4-(aminomethyl)benzyl)amino)methyl)-2-methoxybenzyl)-2-ethoxy-9H-purin-8-ol (Compound 105)

Chemical Structure

[0427] Example 1.18: 6-Amino-9-(4-(((4-aminobutyl)amino)methyl)-2-methoxybenzyl)-2-ethoxy-9H-purin-8-ol (Compound 106)

Chemical Structure

[0428] Example 1.19: 6-Amino-9-(4-((benzylamino)methyl)-2-methoxybenzyl)-2-ethoxy-9H-purin-8-ol (Compound 107)

Chemical Structure

[0429] Example 1.20: 6-Amino-9-(4-((cyclobutylamino)methyl)-2-methoxybenzyl)-2-ethoxy-9H-purin-8-ol (Compound 193)

Chemical Structure

[0430] Example 1.21: 6-Amino-9-(4-((cyclobutylamino)methyl)-2-methoxybenzyl)-2-ethoxy-9H-purin-8-ol (Compound 108)

Chemical Structure

[0431] Example 1.22: 6-Amino-9-(4-(((3,3-difluorocyclobutyl)amino)methyl)-2-methoxybenzyl)-2-ethoxy-9H-purin-8-ol (Compound 109)

Chemical formula

[0432] Example 1.23: 6-Amino-9-(4-((4-aminopiperidin-1-yl)methyl)-2-methoxybenzyl)-2-ethoxy-9H-purin-8-ol (Compound 110)

Chem.

[0433] Example 1.24: 6-Amino-9-(4-(((2-aminoethyl)amino)methyl)-2-methoxybenzyl)-2-ethoxy-9H-purin-8-ol (Compound 111)

Chem.

[0434] = 388.3.

Chem.

[0435] Example 1.26: 2-(((S)-2-Amino-2-carboxyethyl)thio)-4-((2-(4-(4-((6-amino-2-ethoxy-8-hydroxy-9H-purin-9-yl)methyl)-3-methoxybenzyl)piperazin-1-yl)ethyl)amino)-4-oxobutanoic acid (Compound 113)

Chemical Structure

[0436] Example 1.27:N 6 -(4-((6-amino-2-ethoxy-8-hydroxy-9H-purin-9-yl)methyl)-3-methoxybenzyl)-L-lysine (Compound 114) [ka] N in aqueous NaHCO3 (0.275 mL, 1.14 M, 12.5 equiv.) α To a solution of -Boc-L-lysine (32 mg, 0.14 mmol, 5.0 equiv.) was added a solution of compound 11 (0.010 g, 0.027 mmol, 1.0 equiv.) in 100 μL of DMA. The resulting solution was heated to 50° C. for 4 h and then acidified with aqueous HCl (5 M). Preparative HPLC purification was performed as described in General Procedure 5, eluting with a gradient of 10 to 45% CH CN + 0.1% TFA / H O + 0.1% TFA. Deprotection was carried out according to General Procedure 2 to give the title compound 114 as a white solid (7.9 mg, 0.010 mmol, 37%). LC-MS: C 22 H 31 Calculated m / z for N7O5 = 473.2, Measured [M+H] + = 474.3. 11H NMR (300 MHz, MeOD) δ 7.18 - 7.04 (m, 2H), 7.00 (d, J = 7.7 Hz, 1H), 5.05 (s, 2H), 4.34 (q, J = 7.1 Hz, 2H), 4.20 (s, 2H), 3.98 (t, J = 6.3 Hz, 1H), 3.93 (s, 3H), 3.13 - 3.01 (m, 2H), 2.05 - 1.48 (m, 6H), 1.34 (t, J = 7.1 Hz, 4H).

[0437] Example 1.28: 6 - ((4 - ((6 - Amino - 2 - ethoxy - 8 - hydroxy - 9H - purin - 9 - yl)methyl)-3 - methoxybenzyl)-amino)hexanoic acid (Compound 115) [Chemical formula] A solution of Compound 11 (0.010 g, 0.014 mmol, 1.0 equiv) in 100 μL of DMA was added to a solution of 6 - aminohexanoic acid (18 mg, 0.14 mmol, 5.0 equiv) in aqueous NaHCO3 (0.275 mL, 1.14 M, 12.5 equiv). The resulting solution was heated at 50 °C for 4 h and then acidified with aqueous HCl (5 M). Preparative HPLC purification was carried out with a gradient of 10 to 45% CH3CN + 0.1% TFA / H2O + 0.1% TFA for elution as described in General Procedure 5 to give the title compound 115 as a white solid (6.7 mg, 0.013 mmol, 48%). LC - MS: C 22 H 30 Calculated m / z = 458.2 for C21H28N6O5, found [M + H] + = 459.2. 11H NMR (300 MHz, MeOD) δ 7.14 (d, J = 7.5 Hz, 2H), 7.01 (dd, J = 7.6, 1.6 Hz, 1H), 5.06 (s, 2H), 4.46 - 4.33 (m, 2H), 4.19 (s, 2H), 3.93 (s, 3H), 3.11 - 2.99 (m, 2H), 2.34 (t, J = 7.2 Hz, 2H), 1.79 - 1.61 (m, 4H), 1.52 - 1.25 (m, 5H).

[0438] Example 1.29: (3-((4-((6-Amino-2-ethoxy-8-hydroxy-9H-purin-9-yl)methyl)-3-methoxybenzyl)-amino)-propanoyl)-L-histidine (Compound 116)

Chem.

[0439] Example 1.30: (4-((6-Amino-2-ethoxy-8-hydroxy-9H-purin-9-yl)methyl)-3-methoxybenzyl)-L-phenylalanine (Compound 117)

Chemical Structure

[0440] Example 1.31: (S)-6-Amino-2-ethoxy-9-(4-(((1-hydroxy-3-phenylpropan-2-yl)amino)methyl)-2-methoxybenzyl)-9H-purin-8-ol (Compound 118) [Chemical Structure] A solution of compound 11 (0.010 g, 0.014 mmol, 1.0 equivalent) in 100 μL of DMA was added to a solution of L-phenylalaninol (21 mg, 0.14 mmol, 5.0 equivalents) in aqueous NaHCO3 solution (0.275 mL, 1.14 M, 12.5 equivalents). The resulting solution was heated at 50 °C for 4 hours and then acidified with aqueous HCl solution (5 M). Preparative HPLC purification was carried out using a gradient of 10 to 45% CH3CN + 0.1% TFA / H2O + 0.1% TFA for elution as described in General Procedure 5 to obtain the title compound 118 as a white solid (11 mg, 0.015 mmol, 55%). LC-MS: C 25 H 30 Calculated m / z = 478.2 for C24H27N6O4, found [M+H] + = 479.3. 11H NMR (300 MHz, MeOD) δ 7.41 - 7.11 (m, 7H), 7.04 (dd, J = 7.8, 1.6 Hz, 1H), 5.07 (s, 2H), 4.45 - 4.22 (m, 4H), 3.93 (s, 3H), 3.78 (dd, J = 12.1, 3.3 Hz, 1H), 3.58 (dd, J = 12.2, 4.7 Hz, 1H), 3.50 - 3.35 (m, 1H), 3.11 (dd, J = 13.5, 4.7 Hz, 1H), 2.99 (dd, J = 13.5, 10.3 Hz, 1H), 1.36 (t, J = 7.1 Hz, 3H).

[0441] Example 1.32: (4 - ((6 - Amino - 2 - ethoxy - 8 - hydroxy - 9H - purin - 9 - yl)methyl) - 3 - methoxybenzyl) - L - serine (Compound 119) [Chemical formula] A solution of compound 11 (0.010 g, 0.014 mmol, 1.0 equivalent) in 100 μL of DMA was added to a solution of L - serine (14 mg, 0.14 mmol, 5.0 equivalents) in aqueous NaHCO3 solution (0.275 mL, 1.14 M, 12.5 equivalents). The resulting solution was heated at 50 °C for 4 hours and then acidified with aqueous HCl solution (5 M). Preparative HPLC purification was carried out with a gradient of 10 to 45% CH3CN + 0.1% TFA / H2O + 0.1% TFA for elution as described in General Procedure 5, and the title compound 119 was obtained as a white solid (8.7 mg, 0.013 mmol, 48%). LC - MS: C 19 H 24 Calculated m / z = 432.2 for C16H20N6O6, found [M + H] + = 433.3. 11H NMR (300 MHz, MeOD) δ 7.22 - 7.12 (m, 2H), 7.04 (dd, J = 7.7, 1.6 Hz, 1H), 5.08 (s, 2H), 4.43 (q, J = 7.1 Hz, 2H), 4.31 (s, 2H), 4.15 - 3.98 (m, 3H), 3.93 (s, 3H), 1.38 (t, J = 7.1 Hz, 3H).

[0442] Example 1.33: 2,2'-((4-((6-Amino-2-ethoxy-8-hydroxy-9H-purin-9-yl)methyl)-3-methoxy-benzyl)azanediyl)diacetic acid (Compound 120)

Chemical Structure

[0443] Example 1.34: (4-((6-Amino-2-ethoxy-8-hydroxy-9H-purin-9-yl)methyl)-3-methoxybenzyl)-L-isoleucine (Compound 121)

Chem.

[0444] Example 1.35: 6-Amino-2-ethoxy-9-(4-((((2S,3S)-1-hydroxy-3-methylpentan-2-yl)amino)methyl)-2-methoxybenzyl)-9H-purin-8-ol (Compound 122)

Chem.

[0445] Example 1.36: 6-Amino-9-(4-(aminomethyl)-2-methoxybenzyl)-2-ethoxy-9H-purin-8-ol (Compound 123)

Chemical Structure

[0446] Example 1.37: 6-Amino-2-ethoxy-9-(4-((ethylamino)methyl)-2-methoxybenzyl)-9H-purin-8-ol (Compound 124)

Chemical Structure

[0447] Example 1.38: 6-Amino-9-(4-((dipropylamino)methyl)-2-methoxybenzyl)-2-ethoxy-9H-purin-8-ol (Compound 125) [Chemical formula] The title compound was prepared from Compound 11 (0.010 g, 0.027 mmol, 1.0 equivalent) and dipropylamine (19 μL, 0.14 mmol, 5.0 equivalents) according to General Procedure 1. Preparative HPLC purification was carried out with a gradient of 10 to 45% CH3CN + 0.1% TFA / H2O + 0.1% TFA for elution as described in General Procedure 5, and the title compound 125 was obtained as a white solid (2.5 mg, 0.0038 mmol, 14%). LC-MS: C 22 H 32 Calculated m / z = 428.3 for C19H26N6O3, found [M+H] + = 429.3. 1 H NMR (300 MHz, MeOD) δ 7.15 (s, 1H), 7.12 - 6.97 (m, 2H), 5.06 (s, 2H), 4.38 - 4.24 (m, 4H), 3.95 (s, 3H), 3.09 (t, J = 8.5 Hz, 4H), 1.92 - 1.65 (m, 4H), 1.33 (t, J = 7.1 Hz, 3H), 1.00 (t, J = 7.3 Hz, 6H).

[0448] Example 1.39: (E)-6-Amino-9-(4-(((4-aminobut-2-en-1-yl)amino)methyl)-2-methoxybenzyl)-2-ethoxy-9H-purin-8-ol (Compound 126) [Chemical formula] The title compound was prepared from Compound 11 (0.010 g, 0.028 mmol, 1.0 equiv) and tert-butyl N-[(2E)-4-aminobut-2-en-1-yl]carbamate (15 mg, 0.082 mmol, 3.0 equiv) according to General Procedure 1. Preparative HPLC purification was carried out with a gradient of 10 to 45% CH3CN + 0.1% TFA / H2O + 0.1% TFA as described in General Procedure 5. Half of the resulting Boc intermediate was left, and deprotection of the remaining material was carried out according to General Procedure 2 to give the title compound 126 as a white solid (3.0 mg, 0.0070 mmol, 26%). LC-MS: C 20 H 27 Calculated m / z for C + H 1 N7O3 = 413.2, found [M+H]

[0449] Example 1.40: 6-Amino-2-ethoxy-9-(2-methoxy-4-((piperidin-3-ylamino)methyl)benzyl)-9H-purin-8-ol (Compound 127)

Chemical Structure

[0450] Example 1.41: 9-(4-(((1S,4S)-2,5-Diazabicyclo[2.2.1]heptan-2-yl)methyl)-2-methoxybenzyl)-6-amino-2-ethoxy-9H-purin-8-ol (Compound 128)

Chemical Structure

[0451] Example 1.42: 6-Amino-9-(4-((azetidin-3-ylamino)methyl)-2-methoxybenzyl)-2-ethoxy-9H-purin-8-ol (Compound 129)

Chemical Structure

[0452] = 400.3.

Chemical Structure

[0453] Example 1.44: 6-Amino-2-ethoxy-9-(2-methoxy-4-(((3-(piperazin-1-yl)propyl)amino)methyl)benzyl)-9H-purin-8-ol (Compound 131)

Chem.

[0454] Example 1.45: 6-Amino-9-(4-((4-(aminomethyl)piperidin-1-yl)methyl)-2-methoxybenzyl)-2-ethoxy-9H-purin-8-ol (Compound 132)

Chemical Structure

[0455] Example 1.46: (S)-6-Amino-2-ethoxy-9-(2-methoxy-4-((pyrrolidin-3-ylamino)methyl)benzyl)-9H-purin-8-ol (Compound 133)

Chem.

[0456] Example 1.47: 6-Amino-2-ethoxy-9-(2-methoxy-4-(((2-(piperazin-1-yl)ethyl)amino)methyl)-benzyl)-9H-purin-8-ol (Compound 134)

Chemical Structure

[0457] Example 1.48: 6-Amino-2-ethoxy-9-(2-methoxy-4-(((2-morpholinoethyl)amino)methyl)benzyl)-9H-purin-8-ol (Compound 135) [ka] The title compound was prepared from compound 11 (0.010 g, 0.028 mmol, 1.0 equiv.) and N-aminoethylmorpholine (0.010 g, 0.09 mmol, 3 equiv.) according to general procedure 1. Preparative HPLC purification was performed as described in general procedure 5, eluting with a gradient of 10 to 45% CHCN+0.1% TFA / HO+0.1% TFA, to afford the title compound 135 as a white solid (3.1 mg, 0.0039 mmol, 28%). LC-MS: C 22 H 31Calculated value for N7O4, m / z = 457.3, measured value [M+H] + = 458.4. 1 H NMR (300 MHz, MeOD) δ 7.16 (d, J = 1.5 Hz, 1H), 7.09 (d, J = 7.7 Hz, 1H), 7.02 (dd, J = 7.7, 1.6 Hz, 1H), 5.05 (s, 2H), 4.32 (q, J = 7.1 Hz, 2H), 4.26 (s, 2H), 3.94 (s, 3H), 3.85 - 3.76 (m, 4H), 3.04 (t, J = 6.4 Hz, 2H), 2.90 - 2.84 (m, 4H), 1.48 (s, 2H), 1.34 (t, J = 7.1 Hz, 3H).

[0458] Example 1.49: 6-Amino-2-ethoxy-9-(4-(((2-hydroxy-3-morpholinopropyl)amino)methyl)-2-methoxybenzyl)-9H-purin-8-ol (Compound 136)

Chemical formula

[0459] Example 1.50: 6-Amino-2-ethoxy-9-(2-methoxy-4-(((2-thiomorpholinoethyl)amino)methyl)-benzyl)-9H-purin-8-ol (Compound 137) [Chemical formula] The title compound was prepared from Compound 11 (0.010 g, 0.028 mmol, 1.0 equivalent) and 2-(thiomorpholin-4-yl)ethanamine (0.010 g, 0.09 mmol, 3 equivalents) according to General Procedure 1. Preparative HPLC purification was carried out with a gradient of 10 to 45% CH3CN + 0.1% TFA / H2O + 0.1% TFA for elution as described in General Procedure 5, and the title compound 137 was obtained as a white solid (3.5 mg, 0.0043 mmol, 31%). LC-MS: C 22 H 31 Calculated m / z = 473.2 for C17H23N7O3S, found [M+H] + = 473.3. 1 1H NMR (300 MHz, MeOD) δ 7.19 - 6.97 (m, 3H), 5.05 (s, 2H), 4.39 - 4.22 (m, 4H), 3.94 (s, 3H), 3.36 - 3.30 (m, 2H), 3.15 - 2.98 (m, 6H), 2.83 (q, J = 4.6 Hz, 4H), 1.34 (t, J = 7.1 Hz, 3H).

[0460] Example 1.51: 9-(4-([4,4'-Bipiperidin]-1-ylmethyl)-2-methoxybenzyl)-6-amino-2-ethoxy-9H-purin-8-ol (Compound 138)

Chem.

[0461] Example 1.52: 9-(4-((3,9-Diazaspiro[5.5]undecan-3-yl)methyl)-2-methoxybenzyl)-6-amino-2-ethoxy-9H-purin-8-ol (Compound 139)

Chem.

[0462] Example 1.53: 6-Amino-9-(4-((4-(2-aminoethyl)piperidin-1-yl)methyl)-2-methoxybenzyl)-2-ethoxy-9H-purin-8-ol (Compound 140)

Chemical Structure

[0463] Example 1.54: 6-Amino-2-ethoxy-9-(4-(((4-(hydroxymethyl)benzyl)amino)methyl)-2-methoxy-benzyl)-9H-purin-8-ol

Chemical Structure

[0464] Example 1.55: 6-Amino-9-(4-(((4-(chloromethyl)benzyl)amino)methyl)-2-methoxybenzyl)-2-ethoxy-9H-purin-8-ol

Chemical Structure

[0465] Example 1.56: 6-Amino-9-(4-(((4-((dipropylamino)methyl)benzyl)amino)methyl)-2-methoxy-benzyl)-2-ethoxy-9H-purin-8-ol (Compound 141)

Chemical Structure

[0466] Example 1.57: 6-Amino-2-ethoxy-9-(2-methoxy-4-(((4-(morpholinomethyl)benzyl)amino)methyl)benzyl)-9H-purin-8-ol (Compound 142)

Chemical Structure

[0467] Example 1.58: tert-Butyl (1-(4-(((4-((6-Amino-2-ethoxy-8-hydroxy-9H-purin-9-yl)methyl)-3-methoxybenzyl)amino)methyl)benzyl)piperidin-4-yl)carbamate (Compound 143)

Chem.

[0468] Example 1.59: 6-Amino-9-(4-(((4-((4-Aminopiperidin-1-yl)methyl)benzyl)amino)methyl)-2-methoxybenzyl)-2-ethoxy-9H-purin-8-ol (Compound 144)

Chemical Structure

[0469] Example 1.60: tert-Butyl 4-(4-(((4-((6-Amino-2-ethoxy-8-hydroxy-9H-purin-9-yl)methyl)-3-methoxybenzyl)amino)methyl)benzyl)piperazine-1-carboxylate

Chem.

[0470] Example 1.61: 6-Amino-2-ethoxy-9-(2-methoxy-4-(((4-(piperazin-1-ylmethyl)benzyl)amino)methyl)benzyl)-9H-purin-8-ol (Compound 145)

Chem.

[0471] Example 1.62: 6-Amino-9-(4-(((4-(((4-(aminomethyl)benzyl)amino)methyl)benzyl)amino)methyl)-2-methoxybenzyl)-2-ethoxy-9H-purin-8-ol (Compound 146)

Chem.

[0471] 1H NMR (300 MHz, MeOD) δ 7.58 (d, J = 4.0 Hz, 8H), 7.13 (s, 1H), 7.07 - 6.94 (m, 2H), 5.03 (s, 2H), 4.36 - 4.21 (m, 10H), 4.18 (s, 2H), 3.92 (s, 3H), 1.32 (t, J = 7.1 Hz, 3H).

[0472] Example 1.63: 6-Amino-9-(4-(((4-((benzylamino)methyl)benzyl)amino)methyl)-2-methoxy-benzyl)-2-ethoxy-9H-purin-8-ol (Compound 147)

Chemical Structure

[0473] Example 1.64: 2,2'-((4-(((4-((6-Amino-2-ethoxy-8-hydroxy-9H-purin-9-yl)methyl)-3-methoxy-benzyl)amino)methyl)benzyl)azanediyl)diacetic acid (Compound 151) [Chem.] A solution of compound 11 (0.010 g, 0.021 mmol, 1.0 equiv) in 100 μL of DMF was added to a solution of iminodiacetic acid (14 mg, 0.10 mmol, 5.0 equiv) in an aqueous NaHCO3 solution (0.272 mL, 1.14 M, 15.0 equiv). The resulting solution was heated at 60 °C for 18 h. Preparative HPLC purification was carried out with elution using a gradient of 10 to 45% CH3CN + 0.1% TFA / H2O + 0.1% TFA as described in General Procedure 5, and the title compound 151 was obtained as a white solid (1.5 mg, 0.0026 mmol, 26%). LC-MS: C 28 H 33 Calculated m / z = 579.2 for C24H31N7O7, found [M + H] + = 580.3. 1 H NMR (300 MHz, MeOD) δ 7.63 - 7.47 (m, 4H), 7.13 (s, 1H), 7.10 - 6.95 (m, 2H), 5.04 (s, 2H), 4.37 - 4.21 (m, 8H), 3.93 (s, 3H), 3.79 (s, 4H), 1.33 (t, J = 7.1 Hz, 3H).

[0474] Example 1.65: 2-(((S)-2-Amino-2-carboxyethyl)thio)-4-((6-(4-(4-((6-amino-2-ethoxy-8-hydroxy-9H-purin-9-yl)methyl)-3-methoxybenzyl)piperazin-1-yl)-6-oxohexyl)amino)-4-oxobutanoic acid (Compound 148) [Chem.] To a solution of compound 100 (0.020 g, 0.048 mmol, 1.0 equiv) in DMF (500 μL) was added 2,5-dioxopyrrolidin-1-yl 6-(2,5-dioxopyrrol-1-yl)hexanoate (16 mg, 0.053 mmol, 1.1 equiv), followed by DIPEA (42 μL, 0.24 mmol, 5.0 equiv). The resulting solution was stirred at room temperature for 18 h, then L-cysteine (12 mg, 0.096 mmol, 2.0 equiv) was added and the resulting solution was stirred at room temperature for 1 h. Then, H2O (200 μL) was added and the reaction mixture was stirred at room temperature for an additional 2 h. Preparative HPLC purification was carried out eluting with a gradient of 5 to 45% CH3CN + 0.1% TFA / H2O + 0.1% TFA as described in General Procedure 5 to give the title compound 148 as a white solid (5.1 mg, 0.0047 mmol, 9.8%). LC-MS: C 33 H 47 Calculated m / z for C + H9N9O9S = 745.3, found [M+H]

[0475] Example 1.66: (19S)-19-Amino-1-(4-(4-((6-amino-2-ethoxy-8-hydroxy-9H-purin-9-yl)methyl)-3-methoxybenzyl)piperazin-1-yl)-16-carboxy-1,14-dioxo-4,7,10-trioxa-17-thia-13-azicosan-20-oic acid (Compound 149)

Chem.

[0476] Example 1.67: Methyl 4-((6-amino-2-ethoxy-8-methoxy-9H-purin-9-yl)methyl)benzoate

Chemical Structure

[0477] Example 1.68: (4 - ((6 - Amino - 2 - ethoxy - 8 - methoxy - 9H - purin - 9 - yl)methyl)phenyl)methanol

Chem.

[0478] Example 1.69: 6 - Amino - 9 - (3 - (chloromethyl)benzyl)-2 - ethoxy - 9H - purin - 8 - ol

Chem.

[0479] Example 1.70: 6 - Amino - 2 - ethoxy - 9 - (4 - (piperazin - 1 - ylmethyl)benzyl)-9H - purin - 8 - ol (Compound 150) [Chemistry] The title compound was prepared from Compound 17 (16 mg, 0.048 mmol, 1.0 equiv) and tert-butyl piperazine-1-carboxylate (28 mg, 0.14 mmol, 3.0 equiv) according to General Procedure 1. Preparative HPLC purification was performed with a gradient of 10 to 45% CH3CN + 0.1% TFA / H2O + 0.1% TFA for elution as described in General Procedure 5. Half of the obtained Boc intermediate was left, and the deprotection of the remaining material was carried out according to General Procedure 2 to obtain the title compound 150 as a white solid (5.1 mg, 0.0070 mmol, 30%). LC-MS: C 19 H 25 Calculated m / z for C17H19N7O2 = 383.2, found [M+H] + = 383.3. 1 H NMR (300 MHz, MeOD) δ 7.45 (d, J = 8.1 Hz, 2H), 7.37 (d, J = 8.1 Hz, 2H), 5.02 (s, 2H), 4.39 (q, J = 7.1 Hz, 2H), 3.80 (s, 2H), 3.31 - 3.25 (m, 4H), 2.91 - 2.82 (m, 4H), 1.38 (t, J = 7.1 Hz, 3H).

[0480] Example 1.71: Methyl 4-((6-amino-2-ethoxy-8-methoxy-9H-purin-9-yl)methyl)-3-fluorobenzoate [Chemistry] A solution of compound 8 (430 mg, 2.0 mmol, 1.0 equiv) and methyl 4-(bromomethyl)-3-fluorobenzoate (0.50 g, 2.0 mmol, 1.0 equiv) in DMF (5 mL) was added CsCO3 (660 mg, 2.0 mmol, 1.0 equiv). The resulting suspension was stirred at room temperature for 18 h and then concentrated in vacuo. Reverse phase flash purification was carried out using a 60 g C18 column eluting with a gradient of 10 to 50% CH3CN + 0.1% TFA / H2O + 0.1% TFA as described in General Procedure 5. The title compound 18 was obtained as a white solid (0.50 g, 1.3 mmol, 66%). LC-MS: C 17 H 18 Calculated m / z for C , H + FN5O4 = 375.1, found [M+H]

[0481] Example 1.72: (4-((6-Amino-2-ethoxy-8-methoxy-9H-purin-9-yl)methyl)-3-fluorophenyl)-methanol

Chemical formula

[0482] Example 1.73: 6-Amino-9-(4-(chloromethyl)-2-fluorobenzyl)-2-ethoxy-9H-purin-8-ol

Chem.

[0483] Example 1.74: 6-Amino-2-ethoxy-9-(2-fluoro-4-(piperazin-1-ylmethyl)benzyl)-9H-purin-8-ol (Compound 152)

Chem.

[0484] Example 1.75: 6-Amino-9-(4-((4-aminopiperidin-1-yl)methyl)-2-fluorobenzyl)-2-ethoxy-9H-purin-8-ol (Compound 153)

Chemical formula

[0485] Example 1.76: 2-Butoxy-9-(tetrahydro-2H-pyran-2-yl)-9H-purin-6-amine

Chem.

[0486] Example 1.77: 8-Bromo-2-butoxy-9-(tetrahydro-2H-pyran-2-yl)-9H-purin-6-amine

Chem.

[0487] Example 1.78: 2-Butoxy-8-methoxy-9-(tetrahydro-2H-pyran-2-yl)-9H-purin-6-amine

Chem.

[0488] Example 1.79: 2-Butoxy-8-methoxy-9-(tetrahydro-2H-pyran-2-yl)-9H-purin-6-amine

Chem.

[0489] Example 1.80: Methyl 4-((6-amino-2-butoxy-8-methoxy-9H-purin-9-yl)methyl)-3-methoxy-benzoate

Chem.

[0490] Example 1.81: (4-((6-Amino-2-butoxy-8-methoxy-9H-purin-9-yl)methyl)-3-methoxyphenyl)-methanol

Chem.

[0491] Example 1.82: 6-Amino-2-butoxy-9-(4-(chloromethyl)-2-methoxybenzyl)-9H-purin-8-ol

Chem.

[0492] Example 1.83: 6-Amino-2-butoxy-9-(2-methoxy-4-(piperazin-1-ylmethyl)benzyl)-9H-purin-8-ol (Compound 154)

Chem.

[0493] Example 1.84: 6-Amino-9-(4-((4-aminopiperidin-1-yl)methyl)-2-methoxybenzyl)-2-butoxy-9H-purin-8-ol (Compound 155)

Chemical Structure

[0494] Example 1.85: 6-Amino-9-(4-((4-(aminomethyl)piperidin-1-yl)methyl)-2-methoxybenzyl)-2-butoxy-9H-purin-8-ol (Compound 156)

Chem.

[0495] Example 1.86: 9-(4-((3,9-Diazaspiro[5.5]undecan-3-yl)methyl)-2-methoxybenzyl)-6-amino-2-butoxy-9H-purin-8-ol (Compound 157) [Chemical Structure] The title compound was prepared from Compound 27 (30 mg, 0.077 mmol, 1.0 equiv) and tert-butyl 3,9-diazaspiro[5.5]undecane-3-carboxylate (58 mg, 0.23 mmol, 3.0 equiv) according to General Procedure 1. Preparative HPLC purification was performed with a gradient of 10 to 55% CH3CN + 0.1% TFA / H2O + 0.1% TFA for elution as described in General Procedure 5. Deprotection was carried out according to General Procedure 2 to obtain the title compound 157 as a white solid (0.010 g, 0.012 mmol, 15%). LC-MS: C 27 H 39 Calculated m / z = 509.3 for C23H34N7O3, found [M+H] + = 510.3. 11H NMR (300 MHz, MeOD) δ 7.15 (s, 1H), 7.03 (q, J = 7.7 Hz, 2H), 5.05 (s, 2H), 4.32 (s, 2H), 4.25 (t, J = 6.6 Hz, 2H), 3.95 (s, 3H), 3.24 - 3.19 (m, 8H), 3.19 - 3.08 (m, 2H), 2.08 - 1.97 (m, 2H), 1.96 - 1.90 (m, 2H), 1.69 (s, 6H), 1.55 - 1.35 (m, 2H), 0.96 (t, J = 7.4 Hz, 3H).

[0496] Example 1.87: 6-Amino-9-(4-((4-(2-Aminoethyl)piperidin-1-yl)methyl)-2-methoxybenzyl)-2-butoxy-9H-purin-8-ol (Compound 158)

Chem.

[0497] Example 1.88: 6-Amino-2-butoxy-9-(4-(((2-hydroxy-2-methylpropyl)amino)methyl)-2-methoxybenzyl)-9H-purin-8-ol (Compound 159)

Chemical Structure

[0498] Example 1.89: 6-Amino-9-(4-(((4-aminobutyl)amino)methyl)-2-methoxybenzyl)-2-butoxy-9H-purin-8-ol (Compound 160)

Chem.

[0499] Example 1.90: 6-Amino-2-butoxy-9-(4-((((2R,3S)-1-hydroxy-3-methylpentan-2-yl)amino)methyl)-2-methoxybenzyl)-9H-purin-8-ol (Compound 161)

Chem.

[0500] Example 1.91: 6-Amino-9-(4-(aminomethyl)-2-methoxybenzyl)-2-butoxy-9H-purin-8-ol (Compound 162)

Chemical Structure

[0501] Example 1.92: 6-Amino-2-butoxy-9-(2-methoxy-4-((methylamino)methyl)benzyl)-9H-purin-8-ol (Compound 163)

Chemical Structure

[0502] Example 1.93: 6-Amino-2-butoxy-9-(4-((cyclobutylamino)methyl)-2-methoxybenzyl)-9H-purin-8-ol (Compound 164)

Chemical Structure

[0503] Example 1.94: 6-Amino-9-(4-((benzylamino)methyl)-2-methoxybenzyl)-2-butoxy-9H-purin-8-ol (Compound 165)

Chem.

[0504] Example 1.95: Methyl 4-((6-amino-2-butoxy-8-methoxy-9H-purin-9-yl)methyl)benzoate

Chem.

[0505] Example 1.96: (4-((6-Amino-2-butoxy-8-methoxy-9H-purin-9-yl)methyl)phenyl)methanol

Chemical Structure

[0506] Example 1.97: 6-Amino-2-butoxy-9-(4-(chloromethyl)benzyl)-9H-purin-8-ol

Chemical Structure

[0507] Example 1.98: 6-Amino-2-butoxy-9-(4-(piperazin-1-ylmethyl)benzyl)-9H-purin-8-ol (Compound 166)

Chemical Structure

[0508] Example 1.99: 2-Butoxy-9-((6-chloropyridin-3-yl)methyl)-8-methoxy-9H-purin-6-amine

Chem.

[0509] Example 1.100: 6-Amino-2-butoxy-9-((6-(piperazin-1-yl)pyridin-3-yl)methyl)-9H-purin-8-ol (Compound 167)

Chem.

[0510] Example 1.101: 9-((6-(3,9-Diazaspiro[5.5]undecan-3-yl)pyridin-3-yl)methyl)-6-amino-2-butoxy-9H-purin-8-ol (Compound 168)

Chem.

[0511] Example 1.102: 9-(4-Bromo-2-methoxybenzyl)-2-butoxy-8-methoxy-9H-purin-6-amine

Chemical Structure

[0512] Example 1.103: 6-Amino-2-butoxy-9-(2-methoxy-4-(piperazin-1-yl)benzyl)-9H-purin-8-ol (Compound 169)

Chemical Structure

[0513] Example 1.104: 6-Amino-9-(4-(4-aminopiperidin-1-yl)-2-methoxybenzyl)-2-butoxy-9H-purin-8-ol (Compound 170)

Chem.

[0514] = 442.4. H NMR (300 MHz, MeOD) δ 6.96 (d, J = 8.3 Hz, 1H), 6.62 (s, 1H), 6.52 (dd, J = 8.4, 2.2 Hz, 1H), 4.96 (s, 2H), 4.31 (t, J = 6.5 Hz, 2H), 3.84 (s, 3H), 3.90 - 3.75 (m, 3H), 2.86 (t, J = 12.4 Hz, 2H), 2.12 - 2.07 (m, 2H), 1.84 - 1.65 (m, 4H), 1.56 - 1.38 (m, 2H), 0.97 (t, J = 7.4 Hz, 3H).Example 1.105: 6-Amino-2-butoxy-9-(2-methoxy-4-(3,9-diazaspiro[5.5]undecan-3-yl)benzyl)-9H-purin-8-ol (Compound 171) [Chemical formula] A solution of Compound 32 (13 mg, 0.030 mmol, 1.0 equiv), tert-butyl 3,9-diazaspiro[5.5]undecane-3-carboxylate (12 mg, 0.046 mmol, 1.5 equiv), KOtBu (14 mg, 0.12 mmol, 4.0 equiv), Pd2dba3 (5.9 mg, 0.0060 mmol, 0.20 equiv), and RuPhos (11 mg, 0.024 mmol, 0.80 equiv) in DMF (0.50 mL) in a crimp-sealable microwave vial was completely degassed and sealed under N2, then heated at 100 °C for 18 h. The reaction mixture was then diluted with EtOAc (25 mL), filtered through celite, and concentrated in vacuo. Preparative HPLC purification was carried out eluting with a gradient of 10 to 60% CH3CN + 0.1% TFA / H2O + 0.1% TFA as described in General Procedure 5 to afford the Boc / 8-MeO intermediate. This intermediate was then dissolved in CH3CN (500 μL) and 6 M HCl (250 μL), and the mixture was heated at 55 °C for 3 h. The reaction mixture was cooled to room temperature and the title product 171 was obtained after lyophilization (6.4 mg, 0.0088 mmol, 29%). LC-MS: C 26 H 37 Calculated m / z for C + H 11H NMR (300 MHz, MeOD) δ 7.20 - 7.10 (m, 2H), 7.02 (dd, J = 8.2, 2.2 Hz, 1H), 5.03 (s, 2H), 4.31 (t, J = 6.5 Hz, 2H), 3.94 (s, 3H), 3.56 (t, J = 5.7 Hz, 4H), 3.29 - 3.23 (m, 4H), 2.08 - 1.85 (m, 8H), 1.81 - 1.65 (m, 2H), 1.56 - 1.38 (m, 2H), 0.97 (t, J = 7.4 Hz, 3H).

[0515] Example 1.106: 3-(4-((6-Amino-2-butoxy-8-methoxy-9H-purin-9-yl)methyl)-3-methoxy-phenyl)prop-2-yn-1-ol [Chemical formula] A solution of compound 32 (380 mg, 0.87 mmol, 1.0 equiv), Pd(OAc)2 (20 mg, 0.087 mmol, 0.10 equiv), triphenylphosphine (110 mg, 0.44 mmol, 0.50 equiv), DIPEA (0.91 mL, 5.2 mmol, 6.0 equiv) and CuI (33 mg, 0.17 mmol, 0.20 equiv) in anhydrous DMF (4 mL) was sealed in a round-bottom flask equipped with a rubber septum and degassed completely with N2. Anhydrous propargyl alcohol (75 μL, 1.3 mmol, 1.5 equiv) was added by syringe, and the reaction mixture was heated at 75 °C for 18 h under N2, then diluted with H2O (50 mL) and extracted with EtOAc (3 × 50 mL). The pooled organic phases were washed with brine (1 × 50 mL), dried over Na2SO4 and concentrated in vacuo to give the crude product. Reverse-phase flash purification was performed using a 60 g C18 column, eluting with a gradient of 10 to 75% CH3CN + 0.1% TFA / H2O + 0.1% TFA as described in General Procedure 5 to give the title product 33 as a dark red solid (130 mg, 0.32 mmol, 36%). LC-MS: C 21 H 25Calculated value for N5O4, m / z = 411.2, measured value [M+H] + = 412.3.

[0516] Example 1.107: 3-(4-((6-Amino-2-butoxy-8-methoxy-9H-purin-9-yl)methyl)-3-methoxyphenyl)propan-1-ol

Chemical Structure

[0517] Example 1.108: 3-(4-((6-Amino-2-butoxy-8-hydroxy-9H-purin-9-yl)methyl)-3-methoxyphenyl)propyl methanesulfonate

Chemical Structure

[0518] Example 1.109: 6-Amino-2-butoxy-9-(2-methoxy-4-(3-(piperazin-1-yl)propyl)benzyl)-9H-purin-8-ol (Compound 173)

Chem.

[0519] Example 1.110: 6-Amino-9-(4-(3-(4-aminopiperidin-1-yl)propyl)-2-methoxybenzyl)-2-butoxy-9H-purin-8-ol (Compound 172)

Chem.

[0520] Example 1.111: 6-Amino-9-(4-(3-(4-aminopiperidin-1-yl)propyl)-2-methoxybenzyl)-2-butoxy-9H-purin-8-ol (Compound 174)

Chem.

[0521] Example 1.112: 6-Amino-9-(4-(3-(4-(2-aminoethyl)piperidin-1-yl)propyl)-2-methoxybenzyl)-2-butoxy-9H-purin-8-ol (Compound 175) [Chemical formula] The title compound was prepared from Compound 35 (0.010 g, 0.020 mmol, 1.0 equivalent) and tert-butyl N-[2-(piperidin-4-yl)ethyl]carbamate (19 mg, 0.081 mmol, 4.0 equivalents) according to General Procedure 8. Preparative HPLC purification was carried out with a gradient of 10 to 60% CH3CN + 0.1% TFA / H2O + 0.1% TFA for elution as described in General Procedure 5 to obtain the Boc / 8-MeO intermediate. Then, this intermediate was dissolved in CH3CN (400 μL) and 6 M HCl (200 μL), and the mixture was heated at 55 °C for 3 hours. The reaction mixture was cooled to room temperature and lyophilized to obtain the title product 175 (1.6 mg, 0.0019 mmol, 9.4%). LC-MS: C 27 H 41 Calculated m / z = 511.3 for C + H 1 N7O3, found [M+H]

[0522] Example 1.113: 6-Amino-2-butoxy-9-(2-methoxy-4-(3-(piperidin-4-ylamino)propyl)benzyl)-9H-purin-8-ol (Compound 176) [Chem.] The title compound was prepared from Compound 35 (0.010 g, 0.020 mmol, 1.0 eq) and 1-Boc-aminopiperidine carbamate (19 mg, 0.081 mmol, 4.0 eq) according to General Procedure 8. Preparative HPLC purification was carried out with a gradient of 10 to 60% CH3CN + 0.1% TFA / H2O + 0.1% TFA for elution as described in General Procedure 5 to obtain the Boc / 8-MeO intermediate. Subsequently, this intermediate was dissolved in CH3CN (400 μL) and 6 M HCl (200 μL), and the mixture was heated at 55 °C for 3 hours. The reaction mixture was cooled to room temperature and lyophilized to obtain the title product 176 (1.5 mg, 0.0018 mmol, 9.1%). LC-MS: C 25 H 37 Calculated m / z for C + H 1 7O3 = 483.3, found [M+H]

[0523] Example 1.114: Ethyl 2-(3-methoxy-4-methylphenyl)acetate [Chem.] A solution of (3-methoxy-4-methylphenyl)acetic acid (5.0 g, 28 mmol, 1.0 eq) in EtOH (30 mL) was added with concentrated H2SO4 (0.3 mL). The resulting mixture was heated to reflux for 1.5 h, then concentrated in vacuo and redissolved in EtOAc (50 mL). The obtained organic solution was extracted with H2O (1×20 mL), then brine (1×10 mL), dried over MgSO4, and concentrated in vacuo to give the crude title product 36 as a yellow oil, which was carried on to the next step without further purification (assumed quantitative yield). LC-MS: C 12 H 16 Calculated m / z = 208.1 for C + H

[0524] Example 1.115: Ethyl 2-(4-(bromomethyl)-3-methoxyphenyl)acetate

Chemical Structure

[0525] Example 1.116: Ethyl 2-(4-((6-amino-2-butoxy-8-methoxy-9H-purin-9-yl)methyl)-3-methoxyphenyl)acetate

Chem.

[0526] Example 1.117: 2-(4-((6-amino-2-butoxy-8-methoxy-9H-purin-9-yl)methyl)-3-methoxyphenyl)ethan-1-ol

Chem.

[0527] Example 1.118: 6-Amino-2-butoxy-9-(4-(2-chloroethyl)-2-methoxybenzyl)-9H-purin-8-ol

Chemical formula

[0528] Example 1.119: 6-Amino-2-butoxy-9-(2-methoxy-4-(2-(piperazin-1-yl)ethyl)benzyl)-9H-purin-8-ol (Compound 180)

Chemical formula

[0529] Example 1.120: 6-Amino-9-(4-(2-(4-aminopiperidin-1-yl)ethyl)-2-methoxybenzyl)-2-butoxy-9H-purin-8-ol (Compound 181)

Chemical Structure

[0530] Example 1.121: 9-(4-(2-(3,9-Diazaspiro[5.5]undecan-3-yl)ethyl)-2-methoxybenzyl)-6-amino-2-butoxy-9H-purin-8-ol (Compound 177)

Chem.

[0531] Example 1.122: 6-Amino-9-(4-(2-(4-(2-aminoethyl)piperidin-1-yl)ethyl)-2-methoxybenzyl)-2-butoxy-9H-purin-8-ol (Compound 178)

Chem.

[0532] Example 1.123: 6-Amino-2-butoxy-9-(2-methoxy-4-(2-(piperidin-4-ylamino)ethyl)benzyl)-9H-purin-8-ol (Compound 179)

Chem.

[0533] Example 1.124: 2-((1-((tert-Butyldiphenylsilyl)oxy)penta-2-yl)oxy)-9-(tetrahydro-2H-pyran-2-yl)-9H-purin-6-amine [Chemical formula] A mixture of compound 2 (5.0 g, 19 mmol, 1.0 equiv), compound 4 (20.3 g, 59.1 mmol, 3.0 equiv) and CsCO3 (6.4 g, 19 mmol, 1.0 equiv) was heated at 170 °C for 18 h. Then, K2CO3 (20 mg) was added and the reaction was stirred at 170 °C for an additional 72 h. The reaction was diluted with DCM (1000 mL), filtered and concentrated in vacuo to give the crude product as a brown solid. Normal-phase flash purification was performed using a 100 g silica column, eluting with a gradient of 0 to 100% EtOAc / hexane as described in General Procedure 5, followed by reverse-phase flash purification using a 60 g C18 column, eluting with a gradient of 10 to 100% CH3CN + 0.1% TFA / H2O + 0.1% TFA as described in General Procedure 5 to give the title product 41 as a white solid (790 mg, 1.4 mmol, 7.2%). LC-MS: C 31 H 41 Calculated m / z = 559.3 for C, Measured value [M+H] + = 560.4

[0534] Example 1.125: 2-((1-((tert-butyldiphenylsilyl)oxy)pentan-2-yl)oxy)-9H-purin-6-amine

Chem.

Claims

1. Equation (I): 【Chemistry 1】 Compounds thereof, their tautomers, protected forms, and / or pharmaceutically acceptable salts (In the formula, X is either O or NH; Y is either N or CH; R １ is an arbitrarily substituted C ２ ~C ６ C optionally substituted with alkyl or branched ３ ~C ８ It is a hydroxyalkyl group; R ２ is H, halogen, optionally substituted C １ ~C ６ alkyl, or optionally substituted C １ ~C ６ alkoxy; R ３ and R ４ These are H, Q-R, and are independent. ５ is or R ３ and R ４ This is, along with N, an arbitrarily substituted C ２ ~C ６ - Heterocycloalkyl ring, optionally substituted C ３ ~C ７ - Heterobicycloalkyl ring, optionally substituted C ６ ~C １０ - Forms a heterospirocycloalkyl ring or an optionally substituted heteroaryl ring; Q is a combination of C, which can be arbitrarily substituted. １ ~C ６ - Alkyl, optionally substituted C ２ ~C ６ - Alkenil or 【Chemistry 2】 And in the formula, ＊ R ５ It is a junction point with, and # is a junction point with N; R ５ H, OH, NR ６ R ７ , arbitrarily substituted C ３ ~C ６ - Cycloalkyl, optionally substituted C ２ ~C ６ - Heterocycloalkyl, optionally substituted C ３ ~C ７ - Heterobicycloalkyl, optionally substituted C ６ ~C １０ - A heterospirocycloalkyl, an optionally substituted aryl, or an optionally substituted heteroaryl; R ６ and R ７ These are H and C, which are substituted independently. １ ~C ６ Alkyl, optionally substituted C １ ~C ６ It is an alkoxycarbonyl or R ６ and R ７ This is, along with N, an arbitrarily substituted C ２ ~C ６ - Heterocycloalkyl ring, optionally substituted C ３ ~C ７ - Heterobicycloalkyl ring, optionally substituted C ６ ~C １０ - Forms a heterospirocycloalkyl ring or an optionally substituted heteroaryl ring; W is an arbitrarily substituted C ３ ~C ６ - Cycloalkyl, optionally substituted C ２ ~C ６ - Heterocycloalkyl, optionally substituted C ３ ~C ７ - Heterobicycloalkyl, optionally substituted C ６ ~C １０ - A heterospirocycloalkyl, an optionally substituted aryl, or an optionally substituted heteroaryl; n is an integer from 1 to 4; m is an integer from 0 to 4; a and b are each independent integers from 0 to 4. However, R ２ If it is H, (i) R １ This is C, which is arbitrarily substituted in the branch. ３ ~C ８ Is it a hydroxyalkyl group? (ii) X is NH, R １ is an arbitrarily substituted C ５ ~C ６ It is alkyl; or (iii)m is either 0 or 1, R ３ and R ４ This is N along with an unsubstituted piperazinyl ring or an optionally substituted C. ６ ~C １０ (- Forms a heterospirocycloalkyl ring).

2. R ２ is a halogen, optionally substituted C １ ~C ６ Alkyl or optionally substituted C １ ~C ６ The compound according to claim 1, or its tautomer, protected form, and / or pharmaceutically acceptable salt thereof, which is an alkoxy.

3. R ２ C is a halogen or unsubstituted C １ ~C ６ The compound according to claim 1, or its tautomer, protected form, and / or pharmaceutically acceptable salt thereof, which is an alkoxy.

4. R ２ The compound according to claim 1, or its tautomers, protected forms, and / or pharmaceutically acceptable salts thereof, wherein is fluorine or methoxy.

5. The compound according to claim 1, or its tautomers, protected forms, and / or pharmaceutically acceptable salts thereof, wherein X is O.

6. R ３ and R ４ These are independently H, or Q-R ５ The compound according to claim 1, or its tautomer, protected form, and / or pharmaceutically acceptable salt.

7. The compound according to claim 1, or its tautomer, protected form, and / or pharmaceutically acceptable salt thereof, wherein n is 1.

8. The compound according to claim 1, or its tautomers, protected forms, and / or pharmaceutically acceptable salts thereof, wherein m is an integer from 1 to 3.

9. The compound is selected from any one of the compounds 100 to 195 listed in Table 1, the compound according to claim 1, or a tautomer thereof, a protected form, and / or a pharmaceutically acceptable salt thereof.

10. Formula (II): 【Transformation 3】 A compound according to claim 1 having the structure, or a tautomer thereof, a protected form, and / or a pharmaceutically acceptable salt thereof. (In the formula, X is either O or NH; Y is either N or CH; R １ is an arbitrarily substituted C ２ ~C ６ C optionally substituted with alkyl or branched ３ ~C ８ It is a hydroxyalkyl group; R ３ and R ４ are, independently, H, Q-R ５ or R ３ and R ４ together with N, optionally substituted C ２ -C ６ -heterocycloalkyl ring, optionally substituted C ３ -C ７ -heterobicycloalkyl ring, optionally substituted C ６ -C １０ -heterospirocycloalkyl ring or optionally substituted heteroaryl ring; Q is a combination of C, which can be arbitrarily substituted. １ ~C ６ - Alkyl, optionally substituted C ２ ~C ６ - Alkenil or 【Chemistry 4】 And in the formula, ＊ R ５ It is a junction point with, and # is a junction point with N; R ５ H, OH, NR ６ R ７ , arbitrarily substituted C ３ ~C ６ - Carboxyalkyl, optionally substituted C ３ ~C ６ - Cycloalkyl, optionally substituted C ２ ~C ６ - Heterocycloalkyl, optionally substituted C ３ ~C ７ - Heterobicycloalkyl, optionally substituted C ６ ~C １０ - A heterospirocycloalkyl, an optionally substituted aryl, or an optionally substituted heteroaryl; R ６ and R ７ These are H and C, which are substituted independently. １ ~C ６ Alkyl, optionally substituted C １ ~C ６ It is an alkoxycarbonyl or R ６ and R ７ This is, along with N, an arbitrarily substituted C ２ ~C ６ - Heterocycloalkyl ring, optionally substituted C ３ ~C ７ - Heterobicycloalkyl ring, optionally substituted C ６ ~C １０ - Forms a heterospirocycloalkyl ring or an optionally substituted heteroaryl ring; W is an arbitrarily substituted C ３ ~C ６ - Cycloalkyl, optionally substituted C ２ ~C ６ - Heterocycloalkyl, optionally substituted C ３ ~C ７ - Heterobicycloalkyl, optionally substituted C ６ ~C １０ - A heterospirocycloalkyl, an optionally substituted aryl, or an optionally substituted heteroaryl; m is an integer from 0 to 4; a and b are independent integers between 0 and 4.

11. The aforementioned compound, 【Transformation 5】 The compound according to claim 10, or its tautomer, protected form, and / or pharmaceutically acceptable salt.

12. The aforementioned compound, 【Transformation 6】 The compound according to claim 1, or its tautomer, protected form, and / or pharmaceutically acceptable salt.

13. Formula (III): 【Transformation 7】 A compound according to claim 1 having the structure, or a tautomer thereof, a protected form, and / or a pharmaceutically acceptable salt thereof. (In the formula, R １ａ is an arbitrarily substituted C ２ ~C ６ C optionally substituted with alkyl or branched ３ ~C ８ It is a hydroxyalkyl group; R ２ａ is a halogen, optionally substituted C １ ~C ６ Alkyl or optionally substituted C １ ~C ６ It is an alkoxy; R ３ａ and R ４ａ These are H, Q-R, and are independent. ５ａ is or R ３ａ and R ４ａ This is, along with N, an arbitrarily substituted C ２ ~C ６ - Heterocycloalkyl ring, optionally substituted C ３ ~C ７ - Heterobicycloalkyl ring, optionally substituted C ６ ~C １０ - Forms a heterospirocycloalkyl ring or an optionally substituted heteroaryl ring; Q is a combination of C, which can be arbitrarily substituted. １ ~C ６ - Alkyl, optionally substituted C ２ ~C ６ - Alkenil or 【Transformation 8】 And in the formula, ＊ R ５ It is a junction point with, and # is a junction point with N; R ５ａ H, OH, NR ６ａ R ７ａ , arbitrarily substituted C ３ ~C ６ - Carboxyalkyl, optionally substituted C ３ ~C ６ - Cycloalkyl, optionally substituted C ２ ~C ６ - Heterocycloalkyl, optionally substituted C ３ ~C ７ - Heterobicycloalkyl, optionally substituted C ６ ~C １０ - A heterospirocycloalkyl, an optionally substituted aryl, or an optionally substituted heteroaryl; R ６ａ and R ７ａ These are H and C, which are substituted independently. １ ~C ６ Alkyl, optionally substituted C １ ~C ６ It is an alkoxycarbonyl or R ６ａ and R ７ａ This is, along with N, an arbitrarily substituted C ２ ~C ６ - Heterocycloalkyl ring, optionally substituted C ３ ~C ７ - Heterobicycloalkyl ring, optionally substituted C ６ ~C １０ - Forms a heterospirocycloalkyl ring or an optionally substituted heteroaryl ring; W is an arbitrarily substituted C ３ ~C ６ - Cycloalkyl, optionally substituted C ３ ~C ６ - Heterocycloalkyl, optionally substituted C ５ ~C ７ - Heterobicycloalkyl, optionally substituted C ６ ~C １０ - A heterospirocycloalkyl, an optionally substituted aryl, or an optionally substituted heteroaryl; u is an integer between 1 and 3; v is an integer between 0 and 4; a and b are independent integers between 0 and 4.

14. Formula (IV): 【Chemistry 9】 A compound according to claim 13 having the structure, or a tautomer thereof, a protected form, and / or a pharmaceutically acceptable salt thereof. (In the formula, R １ａ is an unsubstituted C ２ ~C ６ Alkyl or branched, unsubstituted C ３ ~C ８ It is a hydroxyalkyl group; R ３ａ and R ４ａ These are H, Q-R, and are independent. ５ａ is or R ３ａ and R ４ａ This is, along with N, an arbitrarily substituted C ２ ~C ６ - Heterocycloalkyl ring, optionally substituted C ３ ~C ７ - Heterobicycloalkyl ring, optionally substituted C ６ ~C １０ - Forms a heterospirocycloalkyl ring or an optionally substituted heteroaryl ring; Q is a combination of C, which can be arbitrarily substituted. １ ~C ６ - Alkyl, optionally substituted C ２ ~C ６ - Alkenil or 【Chemistry 10】 And in the formula, ＊ R ５ It is a junction point with, and # is a junction point with N; R ５ａ H, OH, NR ６ａ R ７ａ , arbitrarily substituted C ３ ~C ６ - Carboxyalkyl, optionally substituted C ３ ~C ６ - Cycloalkyl, optionally substituted C ２ ~C ６ - Heterocycloalkyl, optionally substituted C ３ ~C ７ - Heterobicycloalkyl, optionally substituted C ６ ~C １０ - A heterospirocycloalkyl, an optionally substituted aryl, or an optionally substituted heteroaryl; R ６ａ and R ７ａ These are H and C, which are substituted independently. １ ~C ６ Alkyl, optionally substituted C １ ~C ６ It is an alkoxycarbonyl or R ６ａ and R ７ａ This is, along with N, an arbitrarily substituted C ２ ~C ６ - Heterocycloalkyl ring, optionally substituted C ３ ~C ７ - Heterobicycloalkyl ring, optionally substituted C ６ ~C １０ - Forms a heterospirocycloalkyl ring or an optionally substituted heteroaryl ring; W is an arbitrarily substituted C ３ ~C ６ - Cycloalkyl, optionally substituted C ２ ~C ６ - Heterocycloalkyl, optionally substituted C ３ ~C ７ - Heterobicycloalkyl, optionally substituted C ６ ~C １０ - A heterospirocycloalkyl, an optionally substituted aryl, or an optionally substituted heteroaryl; v is an integer between 0 and 4; a and b are independent integers between 0 and 4.

15. A pharmaceutical composition comprising a compound according to any one of claims 1 to 14, or a tautomer thereof, a protected form, and / or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier or diluent.

16. A pharmaceutical agent for the treatment of cancer comprising a compound according to any one of claims 1 to 14, or a tautomer thereof, a protected form, and / or a pharmaceutically acceptable salt thereof.

17. Formula (A): (A) L-(C) ｐ Compounds containing linker structures (In the formula, L is the linker; C is a compound according to any one of claims 1 to 14; p is an integer between 1 and 5.

18. The compound-linker construct according to claim 17, wherein L is a protease-cleavable linker.

19. Formula (X): (X) T-[L-(C) ｐ ] ｒ complex (In the formula, T is the targeting part, L is the linker; C is a compound according to any one of claims 1 to 14; p is an integer from 1 to 5; r has a value ranging from approximately 1 to approximately 8.

20. The complex according to claim 19, wherein T is an antibody or an antigen-binding antibody fragment.

21. A pharmaceutical composition comprising the complex described in claim 19, and a pharmaceutically acceptable carrier or diluent.

22. A pharmaceutical agent for the treatment of cancer, comprising the complex described in claim 19.

Images