Immunomodulator purine derivative compounds, conjugates thereof, and methods of use thereof

Abstract

Disclosed are purine derivative compounds of general formula (I) wherein X is O or NH; y is CH or N; r1 is an optionally substituted alkyl group; n is 1 to 4; m is 0 to 4, and R2 is H, halogen, alkyl or alkoxy; each of R3 and R1 is H, OH, NR6R7, optionally substituted alkyl, alkenyl, cycloalkyl, heterocycloalkyl, aryl or heteroaryl; or R3, R4 and N together may form a ring. Also disclosed are immunostimulatory antibody conjugates (ISACs) comprising such compounds, and methods for producing and using such compounds, for example, in the treatment of diseases such as cancer. # imgabs0 #

Description

Technical Field

[0001] The present disclosure relates to the field of therapeutics, and in particular to purine analogs, conjugates comprising such purine derivative compounds, their use for modulating immune responses in cells or subjects, and the use of such compounds and conjugates for treating diseases (e.g., cancer, inflammation, etc.). Background Art

[0002] Immunostimulatory compounds and immunoconjugates containing such compounds (e.g., antibody-drug conjugates (ADCs), such as immunostimulatory antibody conjugates (ISACs)) have previously been evaluated for the treatment of various diseases, including cancer. However, such compounds and conjugates remain clinically unproven, primarily due to dose-limiting cytotoxicity (e.g., cytokine release syndrome (CRS)), which may restrict their use to doses that do not provide adequate therapeutic efficacy.

[0003] In addition, due to the highly suppressive tumor microenvironment (TME) and the lack of associated effector immune cells (e.g., T cells), current immunotherapies lack specific efficacy against solid tumors. Compounds that agonize Toll-like receptor 7 (TLR7) are immunostimulatory compounds that can locally "reprogram" the TME at the tumor site; however, serious tolerability issues remain unresolved, which limits the systemic administration of these compounds.

[0004] Although clinical data support the use of TLR7 agonists in oncology, such treatment requires topical application. For example, the TLR7 agonist imiquimod has been approved for topical use in skin oncology applications (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 lentigo maligna) are equally effective when applied topically 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 can lead to dose-limiting toxicities below the effective dose (see, Dudek et al., Clin Cancer Res 2007; 13: 7119-7125). In addition, overexpression and / or activation of some TLRs (including TLR7) can lead to conflicting anti-tumor / pro-tumor activities, in some cases exacerbating rather than reducing inflammation, tumor growth, cell survival, metastasis, and upregulation of pro-inflammatory cytokines (see, Kaczanowska S, et al., J Leukoc Biol. 2013; 93(6): 847-863). Summary of the Invention

[0006] Disclosed herein are purine derivative compounds, such as compounds having a structure according to any one of Formulas (I)-(IV) herein, and conjugates comprising such compounds, such as a conjugate according to Formula (X). Furthermore, the present disclosure discloses methods of producing the compounds and conjugates described herein and their use, for example, in treating diseases such as cancer.

[0007] As further described herein, in vitro agonism of TLR7 using the compounds and conjugates of the present disclosure has been demonstrated by incubating TLR7-expressing immune cells (e.g., PBMCs) with the compounds and conjugates of the present disclosure and measuring one or more downstream effects of TLR7 agonism (such as cytokine induction) (see, e.g., Example 3). In addition, in vivo immunostimulatory antibody-drug conjugate (ISAC) therapy utilizing an antibody targeting a tumor-associated antigen (e.g., Her2) conjugated to at least one of the disclosed compounds can produce a potent anti-tumor response as demonstrated by inhibition of tumor growth rate and / or reduction in tumor volume (see, e.g., Example 6). Surprisingly, a significant reduction in off-target effects—and therefore improved tolerability—was observed during in vivo testing of certain ISACs containing the TLR7 agonistic compounds of the present disclosure compared to the use of conventional immunostimulatory compounds (see, e.g., Example 7).

[0008] More specifically, in various embodiments, the present disclosure relates to compounds according to formula (I):

[0009]

[0010] its tautomers, protected forms and / or pharmaceutically acceptable salts,

[0011] in:

[0012] X is O or NH;

[0013] Y is N or CH;

[0014] R 1 is an optionally substituted C2-C6 alkyl group or a branched and optionally substituted C3-C8 hydroxyalkyl group;

[0015] R 2 is H, halogen, optionally substituted C1-C6 alkyl, or optionally substituted C1-C6 alkoxy;

[0016] R 3 and R 4 H, QR independently 5 , or R 3 and R 4 Together with N, it forms an optionally substituted C2-C6-heterocycloalkyl ring, an optionally substituted C3-C7-heterobicycloalkyl ring, an optionally substituted C6-C 10 - a heterospirocycloalkyl ring or an optionally substituted heteroaryl ring;

[0017] Q is a bond, optionally substituted C1-C6-alkyl, optionally substituted C2-C6-alkenyl or Where * is the same as R 5 's attachment point, and # is the attachment point with N;

[0018] R 5 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;

[0019] 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, it forms an optionally substituted C2-C6-heterocycloalkyl ring, an optionally substituted C3-C7-heterobicycloalkyl ring, an optionally substituted C6-C 10 - a heterospirocycloalkyl ring or an optionally substituted heteroaryl ring;

[0020] 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, optionally substituted aryl or optionally substituted heteroaryl;

[0021] n is an integer from 1 to 4;

[0022] m is an integer from 0 to 4; and

[0023] a and b are independently integers from 0 to 4,

[0024] The condition is that when R 2 When it is H, then:

[0025] (i)R 1 is a branched and optionally substituted C3-C8 hydroxyalkyl group;

[0026] (ii) X is NH and R 1 is an optionally substituted C5-C6 alkyl group; and / or

[0027] (iii) m is 0 or 1, and R 3 and R 4 Together with N, it forms an unsubstituted piperazinyl ring or an optionally substituted C6-C 10 -heterospirocycloalkyl ring.

[0028] In some embodiments, the compound of Formula (I) may have a structure according to Formula (II):

[0029]

[0030] its tautomers, protected forms and / or pharmaceutically acceptable salts,

[0031] in:

[0032] X is O or NH;

[0033] Y is N or CH;

[0034] R 1 is an optionally substituted C2-C6 alkyl group or a branched and optionally substituted C3-C8 hydroxyalkyl group;

[0035] R 3 and R 4 H, QR independently 5 , or R 3 and R 4 Together with N, it forms an optionally substituted C2-C6-heterocycloalkyl ring, an optionally substituted C3-C7-heterobicycloalkyl ring, an optionally substituted C6-C 10 - a heterospirocycloalkyl ring or an optionally substituted heteroaryl ring;

[0036] Q is a bond, optionally substituted C1-C6-alkyl, optionally substituted C2-C6-alkenyl or Where * is the same as R 5 's attachment point, and # is the attachment point with N;

[0037] R 5 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;

[0038] 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, it forms an optionally substituted C2-C6-heterocycloalkyl ring, an optionally substituted C3-C7-heterobicycloalkyl ring, an optionally substituted C6-C 10 - a heterospirocycloalkyl ring or an optionally substituted heteroaryl ring;

[0039] 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, optionally substituted aryl or optionally substituted heteroaryl;

[0040] m is an integer from 0 to 4; and

[0041] a and b are independently integers from 0 to 4.

[0042] In various embodiments, the present disclosure relates to compounds of formula (III):

[0043]

[0044] its tautomers, protected forms and / or pharmaceutically acceptable salts,

[0045] in:

[0046] R 1a is an optionally substituted C2-C6 alkyl group or a branched and optionally substituted C3-C8 hydroxyalkyl group;

[0047] R 2a is halogen, optionally substituted C1-C6 alkyl, or optionally substituted C1-C6 alkoxy;

[0048] R 3a and R 4a H, QR independently 5a , or R3a and R 4a Together with N, it forms an optionally substituted C2-C6-heterocycloalkyl ring, an optionally substituted C3-C7-heterobicycloalkyl ring, an optionally substituted C6-C 10 - a heterospirocycloalkyl ring or an optionally substituted heteroaryl ring;

[0049] Q is a bond, optionally substituted C1-C6-alkyl, optionally substituted C2-C6-alkenyl or Where * is the same as R 5 's attachment point, and # is the attachment point with N;

[0050] R 5a 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;

[0051] R 6a and R 7a are independently H, optionally substituted C1-C6 alkyl, optionally substituted C1-C6 alkoxycarbonyl, or R 6a and R 7a Together with N, it forms an optionally substituted C2-C6-heterocycloalkyl ring, an optionally substituted C3-C7-heterobicycloalkyl ring, an optionally substituted C6-C 10 - a heterospirocycloalkyl ring or an optionally substituted heteroaryl ring;

[0052] 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, optionally substituted aryl or optionally substituted heteroaryl;

[0053] u is an integer from 1 to 3;

[0054] v is an integer from 0 to 4; and

[0055] a and b are independently integers from 0 to 4.

[0056] In certain embodiments of the present disclosure, the compound according to formula (III) may have a structure according to formula (IV):

[0057]

[0058] in:

[0059] R 1a is an unsubstituted C2-C6 alkyl group or a branched and unsubstituted C3-C8 hydroxyalkyl group;

[0060] R 3a and R 4a H, QR independently 5a , or R 3a and R 4a Together with N, it forms an optionally substituted C2-C6-heterocycloalkyl ring, an optionally substituted C3-C7-heterobicycloalkyl ring, an optionally substituted C6-C 10 - a heterospirocycloalkyl ring or an optionally substituted heteroaryl ring;

[0061] Q is a bond, optionally substituted C1-C6-alkyl, optionally substituted C2-C6-alkenyl or Where * is the same as R 5 's attachment point, and # is the attachment point with N;

[0062] R 5a 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;

[0063] R 6a and R 7a are independently H, optionally substituted C1-C6 alkyl, optionally substituted C1-C6 alkoxycarbonyl, or R 6a and R 7a Together with N, it forms an optionally substituted C2-C6-heterocycloalkyl ring, an optionally substituted C3-C7-heterobicycloalkyl ring, an optionally substituted C6-C 10 - a heterospirocycloalkyl ring or an optionally substituted heteroaryl ring;

[0064] 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, optionally substituted aryl or optionally substituted heteroaryl;

[0065] v is an integer from 0 to 4; and

[0066] a and b are independently integers from 0 to 4.

[0067] In some embodiments, the disclosure relates to methods of agonizing TLR7, comprising contacting a cell expressing TLR7 with a compound of any one of Formulae (I)-(IV), thereby agonizing TLR7.

[0068] 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 Formulae (I)-(IV), thereby inducing the release of the cytokine from the cell.

[0069] In some embodiments, the present disclosure relates to a method for inhibiting cancer cell proliferation, comprising contacting a cell population comprising cancer cells and immune cells expressing TLR7 with an effective amount of a compound of any one of Formulas (I)-(IV). Such a method may further comprise stimulating TLR7 with a compound of any one of Formulas (I)-(IV), thereby inhibiting cancer cell proliferation.

[0070] In some embodiments, the present disclosure relates to a method for killing cancer cells, comprising contacting a cell population comprising cancer cells and immune cells expressing TLR7 with an effective amount of a compound of any one of Formulas (I)-(IV). Such a method may further comprise stimulating TLR7 with a compound of any one of Formulas (I)-(IV), thereby killing the cancer cells.

[0071] Other embodiments of the present disclosure relate to methods of stimulating an immune response in a subject in need thereof, comprising administering to the subject an effective amount of a compound of any one of Formulas (I)-(IV). In some embodiments, the compound agonizes TLR7 in the subject, thereby stimulating an immune response in the subject.

[0072] Further embodiments of the present disclosure relate to methods of treating cancer in a subject in need thereof, comprising administering to the subject an effective amount of a compound of any one of Formulas (I)-(IV). Such methods may further comprise agonizing TLR7 in the subject, thereby treating the cancer in the subject.

[0073] In various embodiments, the present disclosure relates to conjugates comprising one or more compounds of any one of formulas (I)-(IV). In various embodiments, such conjugates have a structure according to formula (X):

[0074] (X)

[0075] T-[L-(C) p ] r

[0076] in:

[0077] T is the targeting moiety;

[0078] L is the connector;

[0079] C is a compound of any one of formulas (I) to (IV);

[0080] p is an integer from 1 to 5; and

[0081] The value of r is from about 1 to about 8.

[0082] In various embodiments, the targeting moiety is an antibody or antigen-binding fragment thereof.

[0083] The present disclosure also relates to a method of agonizing TLR7, comprising contacting a cell expressing TLR7 with a conjugate of formula (X), thereby agonizing TLR7.

[0084] Further embodiments herein relate to methods of inducing cytokine release from a cell expressing TLR7, the method comprising contacting the cell with a conjugate of formula (X), thereby inducing release of the cytokine from the cell.

[0085] Further embodiments herein relate to methods for inhibiting cancer cell proliferation, comprising contacting a cell population comprising cancer cells and immune cells expressing TLR7 with an effective amount of a conjugate of formula (X). Such methods may further comprise agonizing TLR7 with a compound of the conjugate of formula (X), thereby inhibiting cancer cell proliferation.

[0086] Other embodiments of the present disclosure relate to methods for killing cancer cells, comprising contacting a cell population comprising cancer cells and immune cells expressing TLR7 with an effective amount of a conjugate of formula (X). Such methods may further comprise exciting TLR7 with a compound of the conjugate of formula (X) (e.g., a compound of any one of formulas (I)-(IV) comprised by the conjugate), thereby killing the cancer cells.

[0087] Further embodiments herein relate to methods of stimulating an immune response in a subject in need thereof, comprising administering to the subject an effective amount of a conjugate of Formula (X). In some embodiments, the conjugate agonizes TLR7 in the subject, for example via a compound of Formula (I)-(IV), thereby stimulating an immune response in the subject.

[0088] Further embodiments of the present disclosure relate to methods of treating cancer in a subject in need thereof, comprising administering to the subject an effective amount of a conjugate of Formula (X). In some embodiments, the conjugate agonizes TLR7 in the subject, for example via a compound of Formula (I)-(IV), thereby treating cancer in the subject. BRIEF DESCRIPTION OF THE DRAWINGS

[0089] The embodiments disclosed herein are illustrated by way of example and not limitation in the accompanying drawings. The description and drawings are for illustration and understanding purposes only and are not intended as definitions of the limits of the compounds, conjugates and methods disclosed herein.

[0090] Figure 1 Shown are general synthetic schemes for producing purine derivative compounds (eg, compounds of Formula (I)-(IV)) according to some embodiments of the present disclosure.

[0091] Figure 2 The anti-tumor activity (in the form of tumor growth inhibition and / or tumor volume reduction) of the ISACs tested during in vivo studies in mice following intravenous (iv) administration of 2.5 mg / kg of the respective ISAC or control preparation is shown.

[0092] Figure 3 As a measure of the tolerance of the tested ISACs, Figure 2 Body weight changes of the animals tested in the in vivo studies mentioned above.

[0093] Figures 4A-4G The results are shown using the trastuzumab-based conjugate at different dose levels and compared to the trastuzumab-based conjugate ( Figure 4F ) as well as unconjugated trastuzumab (abbreviated herein as "Tras") and vehicle ( Figure 4G ) were studied for the following tolerability studies: Tras-MT-VK-PABC-Compound 101 ( Figure 4A )、Tras-MT-VK-PABC-compound 106( Figure 4B )、Tras-MT-VC-PABC-compound 154 ( Figure 4C )、Tras-MT-VC-PABC-compound 155( Figure 4D ) and trastuzumab-ME2-compound 100 ( Figure 4E ).

[0094] Figure 5 Shown are some embodiments according to the present disclosure Figure 2 Correlation between tumor growth rate inhibition and murine IL-6 (mIL-6) induction by the ISACs tested is shown in FIG. DETAILED DESCRIPTION

[0095] In various embodiments, the present disclosure relates to immunostimulatory purine derivative compounds having a structure according to any one of formulas (I)-(IV) and capable of exciting TLR7. Also disclosed herein are compound-linker constructs of formula (A), comprising a compound of the present disclosure coupled to a linker moiety. Further disclosed herein are immunostimulatory conjugates of formula (X), comprising one or more of the purine derivative compounds described herein (e.g., those of formulas (I)-(IV)), coupled to a targeting moiety via a linker (e.g., by using a compound-linker construct of formula (A) as described herein). Also disclosed are methods for preparing the compounds, compound-linker constructs, and conjugates, and for using the compounds, compound-linker constructs, and conjugates, for example, to treat cancer.

[0096] definition

[0097] Unless defined otherwise, all technical and scientific terms used herein have the same meanings as commonly understood by one of ordinary skill in the art.

[0098] Unless otherwise indicated, the term "about" as used herein in the context of a value or range generally refers to ±10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, or 1% of the value or range listed or claimed. 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 still other embodiments, the term "about" refers to a variation of approximately ±1% from a given value. It should be understood that such variations are always included in any given value described herein, whether or not specifically mentioned.

[0099] When used in conjunction with the term "comprising" in this document, the use of the word "a / kind" can mean "one", but it also means "one or more", "at least one", and "one or more than one".

[0100] As used herein, the terms "comprising," "having," "including," and "containing," and grammatical variations thereof, are inclusive or open-ended and do not exclude additional, unrecited elements and / or method steps. When used herein in conjunction with a compound, composition, use, or method, the term "consisting essentially of indicates that additional elements and / or method steps may be present, but that these additionalities do not materially affect the manner in which the recited compound, composition, method, or use functions. When used herein in conjunction with a compound, composition, use, or method, the term "consisting of" excludes the presence of additional elements and / or method steps. A compound, composition, use, or method described herein as comprising certain elements and / or steps may also consist essentially of those elements and / or steps in certain embodiments and consist of those elements and / or steps in other embodiments, whether or not those embodiments are specifically mentioned.

[0101] The terms "alkoxycarbonyl" and "alkoxycarbonyl" are used interchangeably herein and refer to the group -C(O)OR, where R is alkyl.

[0102] As used herein, the term "alkyl" refers to a straight-chain or branched saturated hydrocarbon group containing the specified number of carbon atoms. Examples of alkyl groups include, but are not limited to, methyl, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl, isobutyl, tert-butyl, pentyl, isopentyl, tert-pentyl, neopentyl, 1-methylbutyl, 2-methylbutyl, n-hexyl, and the like.

[0103] As used herein, the term "alkylcycloalkyl" refers to an alkyl group as defined herein substituted by a 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" refer to an alkyl group as defined herein substituted by a heterocycloalkyl, aryl or heteroaryl group, respectively, and are 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 may itself be substituted with one or more of -NH, -COH, -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.

[0104] As used herein, the term "amido" refers to the group -C(O)NRR', where R and R' are independently hydrogen, alkyl, aryl, heteroaryl, cycloalkyl, or heterocycloalkyl.

[0105] As used herein, the term "amino" refers to the group -NRR', where R and R' are independently hydrogen, alkyl, aryl, heteroaryl, cycloalkyl, or heterocycloalkyl.

[0106] As used herein, the term "carboxy" refers to the group -C(O)OR, where R is H, alkyl, aryl, heteroaryl, cycloalkyl, or heterocycloheteroalkyl.

[0107] The term "haloalkyl" as used herein refers to an alkyl group, as defined herein, substituted with one or more halogen atoms.

[0108] As used herein, the terms "halogen" and "halo" refer to fluorine (F), bromine (Br), chlorine (Cl), and iodine (I).

[0109] The term "aminoalkyl," as used herein, refers to an alkyl group, as defined herein, substituted with one or more amino groups (eg, one, two, or three amino groups).

[0110] The term "aminoaryl" as used herein refers to an aryl group, as defined herein, substituted with one amino group.

[0111] Unless otherwise defined, the term "aryl" as used herein refers to a 6- to 12-membered monocyclic or bicyclic hydrocarbon ring system in which at least one ring is aromatic. Examples of aryl groups include, but are not limited to, phenyl, naphthyl, 1,2,3,4-tetrahydro-naphthyl, 5,6,7,8-tetrahydro-naphthyl, indanyl, and the like.

[0112] Unless otherwise defined, the term "cycloalkyl" as used herein refers to a monocyclic or bicyclic saturated hydrocarbon containing the specified 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, and the like.

[0113] Unless otherwise defined, the term "heteroaryl" as used herein refers to a 5- to 12-membered monocyclic or bicyclic ring system containing a specified number of carbon atoms (e.g., C3-C7 (including C3 and C7), such as imidazole, thiazole, etc.), wherein at least one ring atom is a heteroatom and at least one ring is aromatic. Examples of heteroatoms include, but are not limited to, O, S, and N. Examples of heteroaryl groups include, but are not limited to, pyridyl, benzofuranyl, pyrazinyl, pyridazinyl, pyrimidinyl, triazinyl, quinolyl, benzoxazolyl, benzothiazolyl, isoquinolyl, quinazolinyl, quinoxalinyl, pyrrolyl, indolyl, etc.

[0114] As used herein, the term "heterocycloalkyl" refers to a monocyclic or bicyclic non-aromatic ring system containing the specified number of carbon atoms (e.g., C2-C6 (inclusive), for example, azetidine, piperazine, etc.) and in which at least one ring atom is a heteroatom (e.g., O, S, or N). The heterocyclyl substituent may be attached via any available ring atom (e.g., a ring carbon or ring nitrogen). Examples of heterocycloalkyl include, but are not limited to, aziridinyl, azetidinyl, piperidinyl, morpholinyl, piperazinyl, pyrrolidinyl, and the like.

[0115] As used herein with reference to alkyl or heteroalkyl ring systems, the term "bicyclic" generally includes fused and spiro ring systems, unless otherwise defined herein.

[0116] As used herein, the terms "hydroxy" and "hydroxyl" refer to the group -OH.

[0117] As used herein, the term "hydroxyalkyl" refers to a linear 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.

[0118] As used herein, the term "alkylthio" refers to a group -SR, wherein 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.

[0119] As used herein, the terms "thio" and "thiol" refer to the group -SH.

[0120] Unless explicitly indicated as "unsubstituted", any alkyl (e.g., any haloalkyl, hydroxyalkyl, aminoalkyl, etc.), cycloalkyl, heterocycloalkyl, spirocycloalkyl, heterospirocycloalkyl, aryl, or heteroaryl mentioned herein should be understood as "optionally substituted", i.e., each such reference includes both unsubstituted and substituted versions of these groups. For example, a reference to "-C1-C6 alkyl" includes unsubstituted -C1-C6 alkyl and -C1-C6 alkyl substituted with one or more substituents further described herein.

[0121] In various embodiments, the substituents of any of formulae (I) to (IV) in any of the embodiments herein may be optionally substituted with one or more groups selected from the group consisting of: -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)-hetero ... In some embodiments, the alkyl, hydroxyalkyl, C1-C4 aminoalkyl, C1-C4 carboxyalkyl, cycloalkyl, heterocycloalkyl, spiroalkyl, heterospiroalkyl, aryl or heteroaryl groups 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 groups.

[0122] In other embodiments, the substituents of any of Formulas (I)-(IV) in any of the embodiments herein may be optionally substituted with one or more groups selected from the group consisting of: -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.

[0123] In yet other embodiments, the substituents of any of Formulas (I)-(IV) in any of the embodiments herein may be optionally substituted with one or more groups 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.

[0124] Generally, a substituent group of any of Formulas (I)-(IV) described herein as "substituted" may include one substituent or multiple substituents up to the full valence of the substitution of the group. For example, a methyl group may include 1, 2, or 3 substituents, and a phenyl group may include 1, 2, 3, 4, or 5 substituents. When a group is substituted with more than one substituent, the substituents may be the same or they may be different.

[0125] Throughout this disclosure, unless otherwise indicated, the following system is used to number ring atoms in the purine moiety of the compounds described herein ( represents a potential point of attachment to one or more substituents):

[0126]

[0127] The terms "subject" and "patient" are used interchangeably herein and refer to an animal in need of treatment. The animal in need of treatment can be a human or a non-human animal, such as a mammal, a bird, or a 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 a non-human primate.

[0128] With respect to a particular result to be achieved, an "effective amount" of a compound or conjugate described herein is an amount sufficient to achieve the desired result. For example, when it comes to killing cancer cells, an "effective amount" of a compound refers to an amount of the compound sufficient to produce the killing effect.

[0129] It is further understood that the affirmative recitation of a feature in one embodiment is a basis for excluding that feature in another embodiment. In particular, where a list of options is presented for a given embodiment or claim, it is understood that one or more options may be deleted from that list and that the shortened list may form an alternative embodiment, whether or not such alternative embodiment is specifically mentioned.

[0130] It is contemplated that any embodiment discussed herein can be implemented by any method, use, or composition disclosed herein, and vice versa.

[0131] Compounds of the present disclosure

[0132] In various embodiments, the present disclosure discloses purine derivative compounds capable of agonizing TLR7.

[0133] In some embodiments, the present disclosure relates to compounds having formula (I):

[0134]

[0135] its tautomers, protected forms and / or pharmaceutically acceptable salts,

[0136] in:

[0137] X is O or NH;

[0138] Y is N or CH;

[0139] R 1 is an optionally substituted C2-C6 alkyl group or a branched and optionally substituted C3-C8 hydroxyalkyl group;

[0140] R 2is H, halogen, optionally substituted C1-C6 alkyl, or optionally substituted C1-C6 alkoxy;

[0141] R 3 and R 4 H, QR independently 5 , or R 3 and R 4 Together with N, it forms an optionally substituted C2-C6-heterocycloalkyl ring, an optionally substituted C3-C7-heterobicycloalkyl ring, an optionally substituted C6-C 10 - a heterospirocycloalkyl ring or an optionally substituted heteroaryl ring;

[0142] Q is a bond, optionally substituted C1-C6-alkyl, optionally substituted C2-C6-alkenyl or Where * is the same as R 5 's attachment point, and # is the attachment point with N;

[0143] R 5 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;

[0144] 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, it forms an optionally substituted C2-C6-heterocycloalkyl ring, an optionally substituted C3-C7-heterobicycloalkyl ring, an optionally substituted C6-C 10 - a heterospirocycloalkyl ring or an optionally substituted heteroaryl ring;

[0145] 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, optionally substituted aryl or optionally substituted heteroaryl;

[0146] n is an integer from 1 to 4;

[0147] m is an integer from 0 to 4; and

[0148] a and b are independently integers from 0 to 4,

[0149] The condition is that when R 2 When it is H, then:

[0150] (i)R 1 is a branched and optionally substituted C3-C8 hydroxyalkyl group;

[0151] (ii) X is NH and R 1 is an optionally substituted C5-C6 alkyl group; and / or

[0152] (iii) m is 0 or 1, and R 3 and R 4 Together with N, it forms an unsubstituted piperazinyl ring or an optionally substituted C6-C 10 -heterospirocycloalkyl ring.

[0153] 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. 2 is halogen or unsubstituted C1-C6 alkoxy. 2 Can be fluoro or methoxy. In some embodiments, R 2 In other embodiments, R 2 It is a methoxy group.

[0154] Disclosed herein are compounds of formula (I) wherein X is O. In other embodiments, in the compound of formula (I), X is NH.

[0155] In some embodiments, in the compound of formula (I), R 1 is an optionally substituted C2-C6 alkyl. 1 In these embodiments, X may be O.

[0156] In yet other embodiments, R 1 is a branched and optionally substituted C3-C8 hydroxyalkyl. 1 It may be a branched and unsubstituted C3-C8 hydroxyalkyl. 1 is a branched unsubstituted C4-C6 hydroxyalkyl group.

[0157] In such embodiments, the compound of formula (I) may be:

[0158]

[0159] In some embodiments, in the compound of formula (I), R 3 and R 4 are independently H or QR 5 In some embodiments, R 3 and R 4 can all be H. In other embodiments, R 3 and R 4 All are QR 5 In R 3 and R 4 All are QR 5 In some embodiments, Q can be an optionally substituted C1-C6-alkyl, and R 5 Can be H or OH. 3 and R 4 All are QR 5 In some embodiments, Q is unsubstituted C1-C6-alkyl, and R 5 For H.

[0160] In other embodiments, in the compounds of formula (I), Q is a bond and R 5 is an unsubstituted C3-C6-carboxyalkyl group.

[0161] In various other embodiments, in the compounds of formula (I), Q is a bond, an optionally substituted C1-C6-alkyl, an optionally substituted C2-C6-alkenyl, or Where * is the same as R 5 's attachment point, and # is the attachment point with N.

[0162] In R 3 and R 4 In some embodiments where all are H, the compound of formula (I) may be:

[0163]

[0164] In R 3 and R 4 All are QR 5 In some embodiments, the compound of formula (I) can be:

[0165]

[0166] In yet other embodiments, in the compounds of Formula (I), R 3 H and R 4 QR 5 In such embodiments, Q may be a bond, and R 5It can be 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, optionally substituted aryl or optionally substituted heteroaryl. In such embodiments, the compound of formula (I) may be selected from the compounds shown in Sub-Table 1C.

[0167] In some embodiments, in the compound of formula (I), R 3 H and R 4 QR 5 , wherein Q is an optionally substituted C1-C6-alkyl group 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 substituted C1-C6-alkyl by 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 substituted C1-C6-alkyl by one or more of -NH2, -CO2H, -OH, carbonyl, halogen, C1-C2 alkyl and C1-C2 hydroxyalkyl. In some embodiments, the compound of formula (I) can be selected from the compounds shown in sub-table 1D.

[0168] In some embodiments, in the compound of formula (I), R 3 H and R 4 QR 5 , wherein Q is an optionally substituted C2-C6-alkenyl group 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.

[0169] In various embodiments, in R 3 and R 4One or more of them is QR 5 In the compound of formula (I), Q is C3-C6-cycloalkyl, C2-C6-heterocycloalkyl, C3-C7-heterobicycloalkyl, C6-C 10 -C1-C6-alkyl substituted with one or more of heterospirocycloalkyl, aryl or heteroaryl. In some of these embodiments, such ring substituents (hereinafter referred to as "W") may be bridging moieties, so that Q is:

[0170]

[0171] in:

[0172] 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, optionally substituted aryl, or optionally substituted heteroaryl; and

[0173] a and b are independently integers from 0 to 4,

[0174] Where * is the same as R 5 's attachment point, and # is the attachment point with N.

[0175] 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 may be selected from the group consisting of:

[0176]

[0177] wherein a and b are independently 1, 2 or 3.

[0178] In some embodiments, in the compound of formula (I), R 3 and R 4 Together with N, it 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. In some embodiments, R 3 and R 4 Together with N, they form a C3-C6-heterocycloalkyl ring, a C3-C6-heterobicycloalkyl ring, a C6-C 10-heterospirocycloalkyl ring or heteroaryl ring, wherein the ring portion 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.

[0179] In some embodiments, R 3 and R 4 Together with N, forms a ring selected from the group consisting of:

[0180]

[0181] in:

[0182] Z is N or CH; and

[0183] R 8 is H, NH2, optionally substituted C1-C6-alkyl, optionally substituted C1-C6-heteroalkyl, optionally substituted C1-C6-aminoalkyl or optionally substituted C2-C6-heterocycloalkyl.

[0184] In some embodiments, in the compound of formula (I), R 3 and R 4 Together with N, forms a ring selected from the group consisting of:

[0185]

[0186] In some embodiments, in the compound of formula (I), R 3 and R 4 Together with N, forms a ring moiety as described herein, and wherein the compound is selected from the compounds shown in Sub-Table 1B.

[0187] In some embodiments, in R 3 and R 4 At least one of them is QR 5 In the compound of formula (I), R 5 Can be NR 6 R 7 In such embodiments, R 6 and R 7 R may independently be H, optionally substituted C1-C6 alkyl or optionally substituted C1-C6 alkoxycarbonyl. 6 and R 7 are all H. In some embodiments, R 6 and R 7 are all optionally substituted C1-C6 alkyl, and wherein R 6 With R 7In such embodiments, R 6 and R 7 All of R may be C1-C6 alkyl substituted by one or more of -NH2, -CO2H, -OH, carbonyl, halogen, C1-C2 alkylthio, C1-C2 alkyl and C1-C2 hydroxyalkyl. 6 and R 7 is a C1-C2 alkyl group substituted with one or more of -NH2, -CO2H, -OH or halogen. 6 H and R 7 is an optionally substituted C1-C6 alkyl. 6 is H, and R 7 It may be a C1-C6 alkyl group substituted by 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.

[0188] In some embodiments, in R 3 and R 4 At least one of them is QR 5 In the compound of formula (I), R 5 Can be NR 6 R 7 , where R 6 and R 7 Together with N, it 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. 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, forms a ring selected from the group consisting of:

[0189]

[0190] In some embodiments, the compound of Formula (I) is selected from the compounds shown in Sub-Table IE.

[0191] In some embodiments, in the compound of Formula (I), n is 1.

[0192] In some embodiments, in the compounds 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 yet other embodiments, m is 3.

[0193] In some embodiments, in the compound of formula (I), R 2 is H and (a) R 1 is a branched and optionally substituted C3-C8 hydroxyalkyl group, and / or (b) X is NH and R 1 is an optionally substituted C5-C6 alkyl group, and / or (c)m is 0 or 1 and R 3 and R 4 Together with N, it forms an unsubstituted piperazinyl ring or an optionally substituted C6-C 10 -heterospirocycloalkyl ring. In some embodiments, R 2 H and R 1 is a branched and optionally substituted C3-C8 hydroxyalkyl. In such embodiments, R 1 Can be a branched and unsubstituted C3-C8 hydroxyalkyl. In other embodiments, R 2 is H, X is NH, and R 1 is an optionally substituted C5-C6 alkyl. In some of those embodiments, R 1 is an unsubstituted C5-C6 alkyl. In yet other embodiments, R 2 can be H, m can be 0 or 1, and R 3 and R 4 Together with N, it can 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 other embodiments, R 3 and R 4 Together with N, it forms an optionally substituted C6-C 10 -heterospirocycloalkyl ring.

[0194] In some embodiments, the compound of Formula (I) is selected from Table 1 herein, including Sub-Tables 1A-1F.

[0195] In certain embodiments of the present disclosure, the compound of formula (I) has the structure of formula (II):

[0196]

[0197] in:

[0198] X is O or NH;

[0199] Y is N or CH;

[0200] R 1 is an optionally substituted C2-C6 alkyl group or a branched and optionally substituted C3-C8 hydroxyalkyl group;

[0201] R 3 and R 4 H, QR independently 5 , or R 3 and R 4 Together with N, it forms an optionally substituted C2-C6-heterocycloalkyl ring, an optionally substituted C3-C7-heterobicycloalkyl ring, an optionally substituted C6-C 10 - a heterospirocycloalkyl ring or an optionally substituted heteroaryl ring;

[0202] Q is a bond, optionally substituted C1-C6-alkyl, optionally substituted C2-C6-alkenyl or Where * is the same as R 5 's attachment point, and # is the attachment point with N;

[0203] R 5 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;

[0204] 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, it forms an optionally substituted C2-C6-heterocycloalkyl ring, an optionally substituted C3-C7-heterobicycloalkyl ring, an optionally substituted C6-C 10 - a heterospirocycloalkyl ring or an optionally substituted heteroaryl ring;

[0205] 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, optionally substituted aryl or optionally substituted heteroaryl;

[0206] m is an integer from 0 to 4; and

[0207] a and b are independently integers from 0 to 4.

[0208] In some embodiments, in the compound of formula (II), X is O and R 1 is an optionally substituted C2-C6 alkyl. In some of these embodiments, R 1 is an unsubstituted C2-C6 alkyl. In some specific embodiments, R 1 It is ethyl or n-butyl.

[0209] In some embodiments, in the compound of formula (II), Y is CH.

[0210] 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 can 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 other embodiments, m is 3.

[0211] In some embodiments, in the compound of formula (II), R 3 and R 4 are H. In other embodiments, R 3 and R 4 All are QR 5 In such embodiments, Q may be an optionally substituted C1-C6-alkyl group, and R 5 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 an unsubstituted C3-C6-carboxyalkyl group.

[0212] In yet other embodiments, in the compound of formula (II), R 3 H and R 4 QR 5 In such embodiments, Q may be a bond, and R 5 It can be 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, optionally substituted aryl or optionally substituted heteroaryl.

[0213] In some embodiments, in the compound of formula (II), R 3 H and R 4 QR 5 , wherein Q is an optionally substituted C1-C6-alkyl group and R5 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 by one or more of the following: 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 by one or more of -NH2, -CO2H, -OH, carbonyl, halogen, C1-C2 alkyl and C1-C2 hydroxyalkyl.

[0214] In some embodiments, in the compound of formula (II), R 3 H and R 4 QR 5 , wherein Q is an optionally substituted C2-C6-alkenyl group 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.

[0215] In some of these embodiments, the compound is:

[0216]

[0217] In various embodiments, in R 3 and R 4 One or more of them is QR 5 In the compound of formula (II), Q is C3-C6-cycloalkyl, C2-C6-heterocycloalkyl, C3-C7-heterobicycloalkyl, C6-C 10 -C1-C6-alkyl substituted with one or more of heterospirocycloalkyl, aryl or heteroaryl. In some of these embodiments, such ring substituents (hereinafter referred to as "W") may be bridging moieties, so that Q is:

[0218]

[0219] in:

[0220] 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, optionally substituted aryl, or optionally substituted heteroaryl; and

[0221] a and b are independently integers from 0 to 4,

[0222] Where * is the same as R 5 's attachment point, and # is the attachment point with N.

[0223] In such embodiments, W is an unsubstituted ring moiety such that Q may be selected from the group consisting of:

[0224]

[0225] wherein a and b are independently 1, 2 or 3.

[0226] In some embodiments, in the compound of formula (II), R 3 and R 4 Together with N, it 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. In some embodiments, R 3 and R 4 Together with N, they form a C2-C6-heterocycloalkyl ring, a C3-C7-heterobicycloalkyl ring, a C6-C 10 -heterospirocycloalkyl ring or heteroaryl ring, wherein the ring portion is 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 3 and R 4 Together with N, forms a ring selected from the group consisting of:

[0227]

[0228] in:

[0229] Z is N or CH; and

[0230] R 8is H, NH2, optionally substituted C1-C6-alkyl, optionally substituted C1-C6-heteroalkyl, optionally substituted C1-C6-aminoalkyl or optionally substituted C3-C6-heterocycloalkyl.

[0231] In some embodiments, in the compound of formula (II), R 3 and R 4 Together with N, forms a ring selected from the group consisting of:

[0232]

[0233] In some embodiments, in R 3 and R 4 At least one of them is QR 5 In the compound of formula (II), R 5 Can be NR 6 R 7 In such embodiments, R 6 and R 7 R may independently be H, optionally substituted C1-C6 alkyl or optionally substituted C1-C6 alkoxycarbonyl. 6 and R 7 are all H. In some embodiments, R 6 and R 7 are all optionally substituted C1-C6 alkyl groups, such that R 6 With R 7 In such embodiments, R 6 and R 7 All of R may be C1-C6 alkyl substituted by one or more of -NH2, -CO2H, -OH, carbonyl, halogen, C1-C2 alkylthio, C1-C2 alkyl and C1-C2 hydroxyalkyl. 6 and R 7 is a C1-C2 alkyl group substituted with one or more of -NH2, -CO2H, -OH and halogen. 6 H and R 7 is an optionally substituted C1-C6 alkyl. 6 H and R 7 It may be a C1-C6 alkyl group substituted by 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.

[0234] In some embodiments, in the compound of formula (II), R 5 It is NR 6 R 7 , where R 6 and R 7 Together with N, it 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. 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, forms a ring selected from the group consisting of:

[0235]

[0236] In some embodiments, the compound of Formula (II) is selected from Sub-Table 1A.

[0237] In some embodiments, the compound of Formula (I) or Formula (II) is:

[0238]

[0239] In various embodiments, the present disclosure relates to compounds of formula (III):

[0240]

[0241] its tautomers, protected forms and / or pharmaceutically acceptable salts,

[0242] in:

[0243] R 1a is an optionally substituted C2-C6 alkyl group or a branched and optionally substituted C3-C8 hydroxyalkyl group;

[0244] R 2a is halogen, optionally substituted C1-C6 alkyl, or optionally substituted C1-C6 alkoxy;

[0245] R 3a and R 4a H, QR independently 5a , or R 3a and R 4a Together with N, it forms an optionally substituted C2-C6-heterocycloalkyl ring, an optionally substituted C3-C7-heterobicycloalkyl ring, an optionally substituted C6-C 10- a heterospirocycloalkyl ring or an optionally substituted heteroaryl ring;

[0246] Q is a bond, optionally substituted C1-C6-alkyl, optionally substituted C2-C6-alkenyl or Where * is the same as R 5 's attachment point, and # is the attachment point with N;

[0247] R 5a 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;

[0248] R 6a and R 7a are independently H, optionally substituted C1-C6 alkyl, optionally substituted C1-C6 alkoxycarbonyl, or R 6a and R 7a Together with N, it forms an optionally substituted C2-C6-heterocycloalkyl ring, an optionally substituted C3-C7-heterobicycloalkyl ring, an optionally substituted C6-C 10 - a heterospirocycloalkyl ring or an optionally substituted heteroaryl ring;

[0249] 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, optionally substituted aryl or optionally substituted heteroaryl;

[0250] u is an integer from 1 to 3;

[0251] v is an integer from 0 to 4; and

[0252] a and b are independently integers from 0 to 4.

[0253] In some embodiments, in the compound of formula (III), R 1a is an optionally substituted C2-C6 alkyl. In such embodiments, R 1a Can be unsubstituted C2-C6 alkyl. In some of these embodiments, R 1a It may be ethyl or n-butyl.

[0254] In some embodiments, in the compound of formula (III), R 2ais halogen or optionally substituted C1-C6 alkoxy. In such embodiments, R 2a It can be halogen or unsubstituted C1-C6 alkoxy. In certain embodiments, R 2a is fluorine or methoxy.

[0255] In some embodiments, in the compound of formula (III), R 3a and R 4a are H. In other embodiments, R 3a and R 4a All are QR 5a In such embodiments, Q may be an optionally substituted C1-C6-alkyl group, and R 5a 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 an unsubstituted C3-C6-carboxyalkyl group.

[0256] In yet other embodiments, in the compound of formula (III), R 3a H and R 4a QR 5a In such embodiments, Q may be a bond, and R 5a It can be 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, optionally substituted aryl or optionally substituted heteroaryl.

[0257] In some embodiments, in the compound of formula (III), R 3a H and R 4a QR 5a , wherein Q is an optionally substituted C1-C6-alkyl group 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 by 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 by one or more of -NH2, -CO2H, -OH, carbonyl, halogen, C1-C2 alkyl and C1-C2 hydroxyalkyl.

[0258] In some embodiments, in the compound of formula (III), R 3a H and R 4a QR 5a , wherein Q is an optionally substituted C2-C6-alkenyl group 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.

[0259] In various embodiments, in R 3a and R 4a One or more of them is QR 5a In the compound of formula (III), Q is C3-C6-cycloalkyl, C2-C6-heterocycloalkyl, C3-C7-heterobicycloalkyl, C6-C 10 -C1-C6-alkyl substituted with one or more of heterospirocycloalkyl, aryl or heteroaryl. In some of these embodiments, such ring substituents (hereinafter referred to as "W") may be bridging moieties, so that Q is:

[0260]

[0261] in:

[0262] 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, optionally substituted aryl, or optionally substituted heteroaryl; and

[0263] a and b are independently integers from 0 to 4, and wherein * is 5 's attachment point, and # is the attachment point with N.

[0264] In some of these embodiments, W is an unsubstituted ring moiety such that Q can be selected from the group consisting of:

[0265]

[0266] wherein a and b are independently 1, 2 or 3.

[0267] In some embodiments, in the compound of formula (III), R 3a and R 4a Together with N, it 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. In some embodiments, R 3a and R 4a Together with N, they form a C2-C6-heterocycloalkyl ring, a C3-C7-heterobicycloalkyl ring, a C6-C 10 -heterospirocycloalkyl ring or heteroaryl ring, wherein the ring portion 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.

[0268] In some embodiments, R 3a and R 4a Together with N, forms a ring selected from the group consisting of:

[0269]

[0270] in:

[0271] Z is N or CH; and

[0272] R 8a is H, NH2, optionally substituted C1-C6-alkyl, optionally substituted C1-C6-heteroalkyl, optionally substituted C1-C6-aminoalkyl or optionally substituted C2-C6-heterocycloalkyl.

[0273] In some embodiments, in the compound of formula (III), R 3a and R 4a Together with N, forms a ring selected from the group consisting of:

[0274]

[0275] In some embodiments, in the compound of formula (III), R 5a NR 6a R 7a In such embodiments, R 6a and R 7a R may independently be H, optionally substituted C1-C6 alkyl or optionally substituted C1-C6 alkoxycarbonyl. 6a and R 7a are H. In other embodiments, R 6a and R 7a are all optionally substituted C1-C6 alkyl groups, wherein R 6a With R 7a In such embodiments, R 6a and R 7a All of R may be C1-C6 alkyl substituted by one or more of -NH2, -CO2H, -OH, carbonyl, halogen, C1-C2 alkylthio, C1-C2 alkyl and C1-C2 hydroxyalkyl. 6a and R 7a is a C1-C2 alkyl group substituted with one or more of -NH2, -CO2H, -OH or halogen. 6a H and R 7a is an optionally substituted C1-C6 alkyl. 6a is H, and R 7a It may be a C1-C6 alkyl group substituted by 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.

[0276] In some embodiments, in R 3a and R 4a At least one of them is QR 5a In the compound of formula (III), R 5a Can be NR 6a R 7a , where R 6a and R 7a Together with N, it 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. 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, forms a ring selected from the group consisting of:

[0277]

[0278] In various embodiments, in the compound of formula (III), u is 1.

[0279] 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 yet other embodiments, v is 3.

[0280] In some embodiments, the compound of Formula (III) is selected from Sub-Table 1A.

[0281] In various embodiments of the present disclosure, the compound of formula (III) herein is a compound according to formula (IV):

[0282]

[0283] in:

[0284] R 1a is an unsubstituted C2-C6 alkyl group or a branched and unsubstituted C3-C8 hydroxyalkyl group;

[0285] R 3a and R 4a H, QR independently 5a , or R 3a and R 4a Together with N, it forms an optionally substituted C2-C6-heterocycloalkyl ring, an optionally substituted C3-C7-heterobicycloalkyl ring, an optionally substituted C6-C 10 - a heterospirocycloalkyl ring or an optionally substituted heteroaryl ring;

[0286] Q is a bond, optionally substituted C1-C6-alkyl, optionally substituted C2-C6-alkenyl or Where * is the same as R 5 's attachment point, and # is the attachment point with N;

[0287] R 5a H, OH, NR 6a R7a , 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;

[0288] R 6a and R 7a are independently H, optionally substituted C1-C6 alkyl, optionally substituted C1-C6 alkoxycarbonyl, or R 6a and R 7a Together with N, it forms an optionally substituted C2-C6-heterocycloalkyl ring, an optionally substituted C3-C7-heterobicycloalkyl ring, an optionally substituted C6-C 10 - a heterospirocycloalkyl ring or an optionally substituted heteroaryl ring;

[0289] 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, optionally substituted aryl or optionally substituted heteroaryl;

[0290] v is an integer from 0 to 4; and

[0291] a and b are independently integers from 0 to 4.

[0292] In some embodiments, in the compound of formula (IV), R 1a is an unsubstituted C2-C6 alkyl group, such as an unsubstituted C2-C4 alkyl group. In other embodiments, R 1a is a branched unsubstituted C3-C8 hydroxyalkyl group.

[0293] In some embodiments, in the compound of formula (IV), R 3a and R 4a are H. In other embodiments, R 3a and R 4a All are QR 5a In such embodiments, Q may be an optionally substituted C1-C6-alkyl group, and R 5a 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 an unsubstituted C3-C6-carboxyalkyl group.

[0294] In yet other embodiments, in the compound of formula (IV), R3a H and R 4a QR 5a In such embodiments, Q may be a bond, and R 5a It can be H, 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.

[0295] In some embodiments, in the compound of formula (IV), R 3a H and R 4a QR 5a , wherein Q is an optionally substituted C1-C6-alkyl group 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 substituted C1-C6-alkyl by 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 substituted C1-C6-alkyl by one or more of -NH2, -CO2H, -OH, carbonyl, halogen, C1-C2 alkyl and C1-C2 hydroxyalkyl.

[0296] In some embodiments, in the compound of formula (IV), R 3a H and R 4a QR 5a , wherein Q is an optionally substituted C2-C6-alkenyl group 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.

[0297] In various embodiments, in R3a and R 4a One or more of them is QR 5a In the compound of formula (IV), Q is C3-C6-cycloalkyl, C2-C6-heterocycloalkyl, C3-C7-heterobicycloalkyl, C6-C 10 -C1-C6-alkyl substituted with one or more of heterospirocycloalkyl, aryl or heteroaryl. In some of these embodiments, such ring substituents (hereinafter referred to as "W") may be bridging moieties, so that Q is:

[0298]

[0299] in:

[0300] 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, optionally substituted aryl, or optionally substituted heteroaryl; and

[0301] a and b are independently integers from 0 to 4, and wherein * is 5 's attachment point, and # is the attachment point with N.

[0302] In such embodiments, W is an unsubstituted ring moiety such that Q may be selected from the group consisting of:

[0303]

[0304] wherein a and b are independently 1, 2 or 3.

[0305] In some embodiments, in the compound of formula (IV), R 3a and R 4a Together with N, it 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. In some embodiments, R 3a and R 4a Together with N, they form a C2-C6-heterocycloalkyl ring, a C3-C7-heterobicycloalkyl ring, a C6-C 10 -heterospirocycloalkyl ring or heteroaryl ring, wherein the ring portion 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 4aTogether with N, forms a ring selected from the group consisting of:

[0306]

[0307] in:

[0308] Z is N or CH; and

[0309] R 8a is H, NH2, optionally substituted C1-C6-alkyl, optionally substituted C1-C6-heteroalkyl, optionally substituted C1-C6-aminoalkyl or optionally substituted C3-C6-heterocycloalkyl.

[0310] In some embodiments, in the compound of formula (IV), R 3a and R 4a Together with N, forms a ring selected from the group consisting of:

[0311]

[0312] In some embodiments, in R 3a and R 4a One or more of them is QR 5a In the compound of formula (IV), R 5a Can be NR 6a R 7a In such embodiments, R 6a and R 7a R may independently be H, optionally substituted C1-C6 alkyl or optionally substituted C1-C6 alkoxycarbonyl. 6a and R 7a are all H. In some embodiments, R 6a and R 7a are all optionally substituted C1-C6 alkyl groups, wherein R 6a With R 7a In such embodiments, R 6a and R 7a It may be a C1-C6 alkyl group substituted with one or more of -NH2, -CO2H, -OH, carbonyl, halogen, C1-C2 alkylthio, C1-C2 alkyl, and C1-C2 hydroxyalkyl. 6a and R 7a is a C1-C2 alkyl group substituted with one or more of -NH2, -CO2H, -OH or halogen. 6a H and R 7a is an optionally substituted C1-C6 alkyl. 6a is H, and R 7aIt may be a C1-C6 alkyl group substituted by 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.

[0313] In some embodiments, in R 3a and R 4a One or more of them is QR 5a In the compound of formula (IV), R 5a Can be NR 6a R 7a , where R 6a and R 7a Together with N, it 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. 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, forms a ring selected from the group consisting of:

[0314]

[0315] In some embodiments, in the compound of Formula (IV), v is 0. In other embodiments, v is 1. In some embodiments, v is 2. In yet other embodiments, v is 3.

[0316] In some embodiments, the compound of Formula (IV) is selected from the compounds shown in Sub-Table 1A.

[0317] In some embodiments, a compound of the present disclosure, such as a compound of any of Formulas (I)-(IV) herein, is selected from Table 1, including Sub-Tables 1A-1F.

[0318] Table 1: Exemplary Purine Derivative Compounds of the Disclosure

[0319]

[0320]

[0321]

[0322]

[0323]

[0324]

[0325]

[0326]

[0327]

[0328]

[0329]

[0330]

[0331]

[0332]

[0333]

[0334]

[0335]

[0336]

[0337]

[0338]

[0339]

[0340]

[0341]

[0342]

[0343]

[0344]

[0345]

[0346]

[0347]

[0348]

[0349]

[0350]

[0351]

[0352]

[0353] It should be understood that throughout this disclosure, references to compounds of formula (I) include, in various embodiments, compounds of formulae (I)-(IV) to the same extent as if embodiments reciting each of these formulae individually were specifically recited.

[0354] In certain embodiments, compounds of formula (I) as described herein may have sufficiently acidic groups, sufficiently basic groups, or both functional groups, and thus react with a variety of organic and inorganic bases, or organic and inorganic acids, to form pharmaceutically acceptable salts. As used herein, the term "pharmaceutically acceptable salt" refers to a salt of a compound of formula (I) that is substantially non-toxic to living organisms. Typical pharmaceutically acceptable salts include those prepared by reacting a compound of formula (I) with a pharmaceutically acceptable mineral acid or organic acid or organic base or inorganic base. Such salts are referred to as acid addition salts and base addition salts.

[0355] Acids commonly used to form acid addition salts are inorganic acids including, but not limited to, hydrochloric acid, hydrobromic acid, hydroiodic acid, sulfuric acid, phosphoric acid; and organic acids including, but not limited to, p-toluenesulfonic acid, methanesulfonic acid, oxalic acid, p-bromobenzenesulfonic 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, octanoate, acrylate, formate, hydrochloride, dihydrochloride, isobutyrate, hexanoate, 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. Pharmaceutically acceptable acid addition salts of particular interest are those formed with mineral acids such as hydrochloric acid and hydrobromic acid, and those formed with organic acids such as maleic acid and methanesulfonic acid.

[0356] Salts of amine groups may also include quaternary ammonium salts in which the amino nitrogen bears a suitable organic group such as an alkyl, lower alkenyl, lower alkynyl or aralkyl moiety.

[0357] Base addition salts include those derived from inorganic bases such as ammonium or alkali or alkaline earth metal hydroxides, carbonates, bicarbonates, etc. Bases that can be used to prepare 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.

[0358] Those skilled in the art will understand that the particular counterion forming part of a pharmaceutically acceptable salt is generally not a critical property so long as the salt as a whole is pharmaceutically acceptable and so long as the counterion does not contribute undesirable properties to the salt as a whole.

[0359] As described herein, compounds of formula (I)-(IV) may contain one or more free amino, hydroxyl, carbonyl (e.g., ketone or aldehyde) and / or carboxylic acid groups. The present disclosure also encompasses protected versions of compounds of formula (I)-(IV) in which the originally free amino, hydroxyl, carbonyl (e.g., ketone or aldehyde) and / or carboxylic acid groups are protected by appropriate protecting groups. The term "protecting group" refers to a chemical group that masks, reduces, or prevents the reactivity of a functional group when attached to a potentially reactive functional group. Typically, protecting groups can be selectively removed as needed during the synthesis process.

[0360] Protecting groups are well known in the art, and various examples are described, for example, in "Protective Groups in Organic Chemistry" (Greene, W. & Wuts, PGM, 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-ethylsulfonyl (TES), trityl, substituted trityl, tosyl, phthalimide, allyloxycarbonyl (Alloc), and 9-fluorenylmethoxycarbonyl (FMOC). Examples of hydroxy protecting groups include, but are not limited to, acetyl, benzyl (Bn), tert-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), triisopropylsiloxymethyl (TOM) and triisopropylsilyl (TIPS). Examples of carbonyl protecting groups include, but are not limited to, acetals, hemiacetals and ketals. Examples of carboxylic acid protecting groups include, but are not limited to, methyl esters, benzyl esters, tert-butyl esters, silyl esters, orthoesters and oxazoline.

[0361] Certain embodiments relate to pharmaceutically acceptable solvates of compounds of formula (I)-(IV). It will be appreciated by those skilled in the art that certain compounds of formula (I)-(IV) may be combined with solvents such as water, methanol, ethanol, or acetonitrile to form pharmaceutically acceptable solvates, such as corresponding hydrates, methanolates, ethanolates, or acetonitrile compounds. Other examples of solvents that can be used to prepare solvates include isopropyl alcohol, dimethyl sulfoxide, ethyl acetate, acetic acid, ethanolamine, and acetone, as well as miscible preparations of solvate mixtures as known to those skilled in the art.

[0362] In various embodiments, the compounds of the present disclosure (e.g., 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., PBMCs). In such embodiments, when immune cells (e.g., PBMCs) are contacted with the compound, the compound can induce the production of, for example, IL6, IFN-α, and / or TNF-α in the cells.

[0363] In some embodiments, the EC of a compound of the present disclosure (e.g., a compound having a structure according to any one of Formulas (I)-(IV)) that agonizes TLR7 50 The EC of any one of Formula (I) to (IV) may be 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. The TLR7 may be human TLR7 or mouse TLR7 or both. In some embodiments, the compound of any one of Formula (I) to (IV) has an EC of about 1 μM or less for agonizing human and / or mouse TLR7. 50 In some embodiments, the compound of any one of Formulas (I)-(IV) has an EC of about 500 nM or less for agonizing human and / or mouse TLR7. 50 In some embodiments, the compound of any one of Formulas (I)-(IV) has an EC value of about 300 nM or less for agonizing human and / or mouse TLR7. 50 In some embodiments, the compound of any one of Formulas (I)-(IV) has an EC value of about 100 nM or less for agonizing human and / or mouse TLR7. 50 In other embodiments, the compound of any one of Formulas (I)-(IV) has an EC of about 50 nM or less for agonizing human and / or mouse TLR7. 50 In yet other embodiments, the compound of any one of Formulas (I)-(IV) has an EC for agonizing human and / or mouse TLR7 of about 25 nM or less. 50 As used herein, EC 50 The values ​​usually refer to the half-maximal effective concentration of the corresponding compound, where EC 50The value of represents the concentration of compound that induces a biological response (eg, TLR7 agonism, stimulation of the immune system, cytokine production by immune cells, and / or killing of tumor cells) that is halfway between baseline and maximum after a defined exposure duration.

[0364] In some embodiments, the present disclosure relates to purine derivative compounds, such as compounds having a structure according to any one of Formulas (I)-(IV), which agonize the EC of TLR7. 50 The present invention relates to a novel nanostructured ... In some embodiments, the TLR7 is a human TLR7.

[0365] The EC of the compounds of the present disclosure (eg, compounds having a structure according to any one of Formulas (I)-(IV)) that agonize TLR7 50 The value can be determined using various methods known in the art, for example, in vitro using a reporter gene assay employing TLR7 reporter cells. 50 Values ​​can be used in vitro using HEK-Blue TM TLR7 reporter cells (eg, available from Invivogen, San Diego, CA) and determined as described, eg, in Example 3 herein.

[0366] In some embodiments, and as further described herein, it has surprisingly been found that, particularly those in R 2Contains a substituent (ie, R 2 Purine derivative compounds of formula (I)-(IV) with ≠H) can provide potent immunomodulatory activity in vitro and in vivo when used as free molecules and in the form of immunostimulatory antibody drug conjugates (ISACs). Such compounds can elicit immunomodulatory activity targeting mouse and human TLR7 in mouse and human environments. In various embodiments, such compounds in R 2 The substituent at position may be an alkoxy moiety or a halogen atom. In a specific embodiment, R 2 The substituents may be methoxy or fluorine. 50 It can be equal to or less than about 500 nM, 100 nM, 50 nM, 30 nM, 20 nM, equal to or less than about 10 nM, or lower, as determined, for example, using a reporter gene assay (RGA) as described herein.

[0367] In other embodiments, wherein R 2 is H and (i) R 1 is a branched and optionally substituted C3-C8 hydroxyalkyl group, (ii) X is NH and R 1 is an optionally substituted C5-C6 alkyl group, or (iii) m is 0 or 1 and R 3 and R 4 Compounds of formula (I) which, together with N, form an unsubstituted piperazinyl group may possess potent immunomodulatory properties useful in the treatment of diseases such as cancer.

[0368] In some embodiments, compounds of the present disclosure (e.g., compounds having a structure according to any one of Formulas (I)-(IV)) can have an EC of 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 to induce production of one or more human and / or murine cytokines by human or murine immune cells, respectively. 50 In some embodiments, the compounds of the present disclosure (e.g., compounds having a structure according to any one of Formulas (I)-(IV)) can have an EC of 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 to induce human or murine immune cells to produce one or more human and / or murine cytokines, respectively. 50In various embodiments, the one or more human and / or murine cytokines include IL6. In some embodiments, the human immune cells include PBMCs and the murine immune cells include murine splenocytes. In various embodiments, an EC that induces PBMCs or mouse splenocytes to produce human or murine IL6 50 Values ​​are determined using, for example, the human PBMC assay or the mouse splenocyte assay, respectively, as described in Example 3 herein.

[0369] In some embodiments, the compounds of the present disclosure (e.g., compounds having a structure according to any one of Formulas (I)-(IV)) can have an EC for inducing the production of human interferon-α (hIFN-α) of about 10 nM or less, 5 nM or less, 1 nM or less, or 0.1 nM or less. 50 In some embodiments, compounds of the present disclosure (e.g., compounds having a structure according to any one of Formulas (I)-(IV)) can have an EC for inducing hIFN-α production of about 10 nM to about 1 nM, about 1 nM to about 0.1 nM, or less than 0.1 nM. 50 In various embodiments, ECs that induce PBMC to produce hIFN-α 50 Values ​​are determined using, for example, the human PBMC assay as described in Example 3 herein.

[0370] In certain embodiments, compounds of the present disclosure (e.g., compounds having a structure according to any one of Formulas (I)-(IV)) induce EC production of cytokines by PBMCs. 50 The ECs are determined in vitro by treating PBMCs isolated from peripheral blood with titrated concentrations of the test compound and subsequently measuring cytokines by, for example, homogeneous time-resolved fluorescence (HTRF). 50 Example method for values.

[0371] The purine derivative compounds of the present disclosure (e.g., those of Formula (I)-(IV)) can be prepared by standard synthetic organic chemistry methods from commercially available starting materials and reagents. Example 1 provided herein details representative examples of suitable synthetic routes (see also Figure 1 Those skilled in the art will recognize that alternative methods may be employed to synthesize the purine derivative compounds of the present disclosure, and therefore the methods described herein are not intended to be exhaustive.

[0372] Compound-linker constructs

[0373] The purine derivative compounds of the present disclosure can be coupled to a linker moiety. In certain embodiments, the present disclosure relates to compound-linker constructs comprising one or more compounds of the present disclosure (C), such as those having a structure according to any one of Formulas (I)-(IV), such as any of the compounds listed in Table 1, coupled to a linker moiety (L).

[0374] In various embodiments, the present disclosure relates to compound-linker constructs having Formula (A):

[0375] (A)

[0376] L-(C) p

[0377] in:

[0378] L is the connector;

[0379] C is a compound of any one of formulas (I)-(IV); and

[0380] p is an integer from 1 to 5.

[0381] In various embodiments of the compound-linker construct, p can be an integer from 1 to 4. In other embodiments, p is an integer from 1 to 3. In still other embodiments, p is an integer from 1 to 2. In various embodiments, p is 1. In other embodiments, p is 2. In still other embodiments, p is 3.

[0382] The linker (L) herein can be a bifunctional or multifunctional moiety capable of linking one or more immunomodulatory purine derivative compounds C of the present disclosure to another molecule (such as a targeting moiety T), as further described elsewhere herein.

[0383] A bifunctional (or monovalent) linker L connects a single compound C to a single site (e.g., a functional group) on a targeting moiety T. In such embodiments, p is 1. A multifunctional (or multivalent) linker L connects more than one compound C to a single site (e.g., a functional group) on a targeting moiety T. Linkers that connect one compound C to more than one site on a targeting moiety T can also be considered multifunctional. However, in embodiments where the linker (L) is multivalent and couples more than one compound (C) to another moiety (such as a targeting moiety (T)), p may be > 1, such as an integer between 2 and 5.

[0384] The linker L of the compound-linker construct herein may comprise a functional group capable of reacting with one or more target groups on the targeting moiety T, and at least one functional group capable of reacting with a target group on the compound of the present disclosure (e.g., a compound according to any one of formulas (I)-(IV)) C. Suitable functional groups are known in the art and include those described herein, such as those described in Bioconjugate Techniques (G.T. Hermanson, 2013, Academic Press). Functional groups on the targeting moiety T and the purine derivative compound of the present disclosure C that can serve as target groups for attachment of the linker include, but are not limited to, thiol, hydroxyl, carboxyl, amine, aldehyde, and ketone groups.

[0385] In some embodiments, the linker herein comprises or consists of a substituted or unsubstituted alkyl or substituted or unsubstituted heteroalkyl chain comprising a first and a second terminal functional group. 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., a targeting moiety, T), and the second terminal functional group of the linker forms a second linkage or bond with a second reactive functional group of a second conjugation partner (e.g., compound C of formula (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.

[0386] In some embodiments, the joint (L) is a cleavable joint. In other embodiments, the joint (L) is a non-cleavable joint. The cleavable joint herein is a joint that is easy to crack under specific conditions, such as intracellular conditions (such as in endosomes or lysosomes) or near target cells (such as in tumor microenvironment). Example includes protease-sensitive, acid-sensitive or reduction-sensitive joints. In contrast, non-cleavable joints can depend on the degradation of the targeting moiety (such as antibody) to which it is attached in the cell, which generally results in the release of the amino acid joint compound part. In some embodiments, the joint includes an oxyalkylene, such as poly (oxyalkylene) or polyethylene glycol (PEG) moiety.

[0387] In some embodiments, the linker (L) of the compound-linker construct herein comprises a polyethylene glycol (PEG) moiety. Such a PEG moiety can have a molecular weight of about 500 Da to about 5 kDa, about 500 Da to about 3 kDa, about 500 Da to about 1 kDa, about 500 Da to about 1 kDa, or about 100 Da to 500 Da.

[0388] The example of cleavable linker includes for example a linker comprising an amino acid sequence as a cleavage recognition sequence of a protease. Many such cleavage recognition sequences are known in the art. In embodiments where the compound-linker construct herein is used in the form of a conjugate not intended to be internalized by the cell, for example, an amino acid sequence recognized and cleaved by the protease present in the extracellular matrix near the target cell (such as a cancer cell) can be used. The example of extracellular tumor-associated protease includes for example plasmin, matrix metalloproteinase (MMP), elastase and kallikrein-related peptidase.

[0389] In embodiments where the compound-linker constructs herein are used as conjugates intended to be internalized by cells, the linker L may comprise an amino acid sequence that is recognized and cleaved by an endosomal or lysosomal protease. Examples of such proteases include, for example, cathepsins B, C, D, H, L, and S, and legumin.

[0390] The cleavage recognition sequence can be, for example, a dipeptide, a tripeptide, or a tetrapeptide. Non-limiting examples of dipeptide recognition sequences that can be included in the cleavable linkers described herein include, but are not limited to, 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. Examples of tripeptide and tetrapeptide cleavage sequences include, but are not limited to, 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. In some embodiments, the linker (L) comprises a dipeptide, a tripeptide, a 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.

[0391] Additional examples of cleavable linkers include disulfide-containing linkers, such as N-succinimidyl-4-(2-pyridyldithio)butyrate (SPDB) and N-succinimidyl-4-(2-pyridyldithio)-2-sulfobutyrate (sulfo-SPDB). Disulfide-containing linkers may optionally include additional groups to provide steric hindrance near the disulfide bond to improve the extracellular stability of the linker, for example, including a geminal dimethyl group. Other cleavable linkers include linkers that are hydrolyzable at a specific pH or pH range, such as hydrazone linkers. Linkers comprising a combination of these functionalities may also be useful, for example, linkers comprising both hydrazone and disulfide bonds are known in the art.

[0392] Another example of a cleavable linker is a linker comprising β-glucuronide, which can be cleaved by β-glucuronidase, an enzyme present in lysosomes and tumor stroma (see, e.g., De Graaf et al., 2002, Curr. Pharm. Des. 8: 1391-1403, and International Patent Publication No. WO 2007 / 011968). β-glucuronide can also be used to improve the hydrophilicity of the linker L.

[0393] Another example of a linker that is cleaved internally within the cell and improves hydrophilicity is a linker comprising a pyrophosphate diester moiety (see, eg, Kern et al., 2016, J Am Chem Soc., 138:2430-1445).

[0394] In certain embodiments, the linker L included in the compound-linker construct of formula (A) is a cleavable linker. In some embodiments, such linker L comprises a cleavage recognition sequence. In some embodiments, such linker L may comprise an amino acid sequence, for example, an amino acid sequence of 2 or 3 residues in length, and may be recognized and cleaved by a protease.

[0395] The cleavable linker may optionally further comprise one or more additional functional groups, such as self-immolative and self-immolative groups, stretchers, hydrophilic moieties, or combinations thereof.

[0396] The self-decomposing and self-eliminating groups in the joints that can be used herein include, for example, p-aminobenzyl (PAB) and p-aminobenzyloxycarbonyl (PABC) groups, and methylated ethylenediamine (MED). Other examples of self-decomposing groups include, but are not limited to, aromatic compounds that are 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. Other examples include groups that undergo cyclization when the amide bond is hydrolyzed, such as substituted and unsubstituted 4-aminobutyric acid amide (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-decomposing / self-eliminating groups are generally attached to amino or hydroxyl groups on compound C. Self-decomposing / self-eliminating groups are often included in peptide-based joints alone or in combination, but may also be included in other types of joints.

[0397] Extenders that can be used in linkers in conjugates as further described herein include, for example, alkylene and aliphatic acid, diacid, amine, or diamine based extenders such as diglycolates, malonates, hexanoates, and hexanamides. Other extenders include, for example, glycine based extenders and polyethylene glycol (PEG) or monomethoxypolyethylene glycol (mPEG) extenders.

[0398] PEG and mPEG extenders can also be used as hydrophilic moieties in the joints of the present disclosure. For example, PEG or mPEG can be "directly inserted" or included in the joint as a side group to increase the hydrophilicity of the joint (see, for example, U.S. Patent Application Publication No. US2016 / 0310612). Various PEG-containing joints are commercially available from companies such as Quanta BioDesign, Ltd (Plain City, OH). Other hydrophilic groups that may be optionally incorporated into the joint L include, for example, β-glucuronide, sulfonate groups, carboxylate groups, and pyrophosphate diesters.

[0399] In some embodiments, the compound-linker constructs of the present disclosure may comprise or consist of a compound of Formula (I)-(IV) herein coupled to any one of the linker moieties L1-L18 shown in Table 2A ( represents attachment to a compound of formula (I)-(IV)).

[0400] Table 2A: Exemplary Linker Moieties of Compound-Linker Constructs

[0401]

[0402]

[0403]

[0404] 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.

[0405] In various embodiments, with reference to Formula (I), the linker moiety (L) can be attached to the compound having the structure of Formula (I) at any suitable atom. In any of the above methods, suitable groups on the compound C of Formulas (I)-(IV) for attaching the linker L include, but are not limited to, thiol groups, amine groups, carboxylic acid groups, and hydroxyl groups. In some embodiments of the present disclosure, the linker L is attached to the compound C of Formula (I) via a hydroxyl or amine group on the compound.

[0406] In some embodiments, the attachment site of the linker moiety is one of the sites shown below in Formula (IL) based on Formula (I), where "L" represents a potential linker attachment site:

[0407]

[0408] “R 3 / 4 " means that when R in Formula I 3 or R 4 When H, such hydrogen atom may be replaced by a linker moiety, such as shown herein for MT-VC-PABC-compound 105 and MT-VC-PABC-compound 107. In addition, as well as in Formula I, R 3 and / or R 4 In embodiments where the substituent contains one or more additional nitrogen atoms suitable for attachment to a linker L, such one or more nitrogen atoms can be the site of attachment of the linker moiety L. Examples include MT-VC-PABC-Compound 100, MT-VC-PABC-Compound 106, and MT-VC-PABC-Compound 111.

[0409] In some embodiments, a compound-linker construct comprising a purine derivative compound of any one of Formulae (I)-(IV) herein can be selected from Table 2B.

[0410] Table 2B: Exemplary Compound-Linker Constructs of the Disclosure

[0411]

[0412]

[0413]

[0414]

[0415]

[0416]

[0417] The compound-linker constructs of Formula (A) herein can be prepared using any suitable synthetic technique known in the art, for example as further described herein in Example 2.

[0418] Conjugates containing purine derivative compounds

[0419] Further disclosed herein are conjugates (also referred to herein as "immunostimulatory antibody-drug conjugates" or "ISACs") comprising a targeting moiety (T) coupled to one or more compounds of the present disclosure (e.g., one or more compounds according to any one or more of Formulas (I)-(IV)). In various embodiments, the conjugates herein comprise one or more of Formulas (A), L-(C), and (D) as described herein. p Compound-linker constructs.

[0420] In some embodiments, the conjugates 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 conjugates of the present disclosure have a structure according to formula (X):

[0421] (X)

[0422] T-[L-(C) p ] r

[0423] in:

[0424] T is the targeting moiety;

[0425] L is the connector;

[0426] C is a compound according to any one of formulae (I)-(IV);

[0427] p is an integer from 1 to 5; and

[0428] The value of r is from about 1 to about 8.

[0429] In various embodiments of the conjugate of formula (X) herein, L-(C) p is a compound-linker construct as described herein.

[0430] As further described herein, in certain embodiments, the compounds disclosed herein can be used to prepare conjugates as immunostimulatory antibody-drug conjugates (ISACs) 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 tumor cell to be targeted). At the target site (e.g., a tumor), one or more immunostimulatory compounds of the present disclosure attached to the targeting moiety can be released into immune cells and / or into the tumor microenvironment (TME), for example, by using a cleavable linker, thereby causing an anti-tumor effect by activating the immune system (e.g., via TLR7-mediated signaling and / or cytokine production).

[0431] A. Targeting moiety (T)

[0432] The targeting moiety T included in the conjugate of formula (X) can be a molecule that binds to, reactively associates with, or complexes with a receptor, an antigen, or other receptive moiety associated with a given target cell population. Typically, the targeting moiety T serves to deliver the purine derivative compound C of the present disclosure to a specific target cell population that reacts with the targeting moiety T. Examples of targeting moieties include, but are not limited to, proteins (such as antibodies, antibody fragments, and growth factors), glycoproteins, peptides (such as bombesin and gastrin-releasing peptide), lectins, vitamins (such as folic acid), and nutrient transport molecules (such as transferrin).

[0433] Typically, the targeting moiety T will be bonded to the linker L via a heteroatom such as sulfur (e.g., from a sulfhydryl group), oxygen (e.g., from a carbonyl, carboxyl, or hydroxyl group), or nitrogen (e.g., from a primary or secondary amino group) of the targeting moiety T. These heteroatoms may be naturally present on the targeting moiety T, or may be introduced by engineering and / or expression, or may be introduced via chemical modification using techniques known in the art.

[0434] In some embodiments, the targeting moiety T is an antibody. Thus, because the compounds of the present disclosure are capable of eliciting an immune response in a subject, various embodiments of the present disclosure are directed to immunostimulatory antibody-drug conjugates (ISACs) having the general formula (X), wherein the targeting moiety T is an antibody.

[0435] In embodiments where the conjugate is an ISAC, the antibody included as the targeting moiety T can be a full-size polyclonal or monoclonal antibody, an antigen-binding antibody fragment (such as Fab, scFab, Fab', F(ab')2, Fv or scFv), a domain antibody (dAb) or an antibody mimetic (such as an affibody, DARPin, anticalin, versabody, duocalin, lipocalin or avimer). Antibodies are typically directed against a specific antigen, such as a tumor-associated antigen (TAA).

[0436] In certain embodiments where the conjugate (X) is an ISAC, the targeting moiety T is a monoclonal antibody, an antigen-binding antibody fragment thereof (such as Fab, scFab, Fab', F(ab')2, Fv or scFv), or a domain antibody (dAb).

[0437] In certain embodiments, the targeting moiety T can be a monoclonal antibody. The monoclonal antibody can be, for example, a non-human monoclonal antibody (such as 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 a targeting moiety in the conjugate (X) herein can be a bispecific antibody or a multispecific antibody.

[0438] In certain embodiments, the targeting moiety T comprised by the conjugate is an antibody or antigen-binding antibody fragment that binds to a tumor-associated antigen (TAA). In various embodiments, such a TAA-binding antibody may comprise a functional Fc region capable of binding to an Fc receptor.

[0439] B. Linkers (L) used in conjugates

[0440] As further described herein, for example, in the "Compound-Linker Constructs" subsection, a linker can couple one or more immunomodulatory compounds of the present disclosure (e.g., any of Compounds (I)-(IV)) to a targeting moiety (T). For a detailed description of linker moieties contemplated herein, please refer to the "Compound-Linker Constructs" subsection above.

[0441] As further described herein, the linker L of the compound-linker construct herein may comprise a functional group capable of reacting with one or more target groups on the targeting moiety T, and at least one functional group capable of reacting with a target group on the compound of the present disclosure (e.g., a compound according to any one of formulas (I)-(IV)) C. Suitable functional groups are known in the art and include those described herein, such as those described in Bioconjugate Techniques (G.T. Hermanson, 2013, Academic Press). Functional groups on the targeting moiety T and the purine derivative compound of the present disclosure C that can serve as target groups for attachment of the linker include, but are not limited to, thiol, hydroxyl, carboxyl, amine, aldehyde, and ketone groups.

[0442] Non-limiting examples of functional groups capable of reacting with thiols include maleimides, haloacetamides, haloacetyl groups, activated esters (such as succinimidyl esters, 4-nitrophenyl esters, pentafluorophenyl esters and tetrafluorophenyl esters), anhydrides, acyl chlorides, sulfonyl chlorides, isocyanates and isothiocyanates. In this case, "self-stabilizing" maleimides as described in Lyon et al., 2014, Nat. Biotechnol., 32: 1059-1062 can also be used.

[0443] Non-limiting examples of functional groups capable of reacting with amines include activated esters (such as N-hydroxysuccinamide (NHS) esters and sulfo-NHS esters), imidoesters (such as Traut's reagent), isothiocyanates, aldehydes, and anhydrides (such as diethylenetriaminepentaacetic anhydride (DTPA). Other examples include succinimidyl-1,1,3,3-tetramethyluronium tetrafluoroborate (TSTU) and benzotriazol-1-yl-oxytripyrrolidinylphosphonium hexafluorophosphate (PyBOP).

[0444] Non-limiting examples of functional groups capable of reacting with an electrophilic group such as an aldehyde or ketone carbonyl include hydrazide, oxime, amino, hydrazine, thiosemicarbazone, hydrazine carboxylate, and arylhydrazide.

[0445] In certain embodiments where the targeting moiety T is an antibody, the linker moiety L may comprise a functional group that allows for bridging of two interchain cysteines on the antibody, such as ThioBridge TM linkers (Badescu et al., 2014, Bioconjug. Chem. 25: 1124-1136), disulfidomaleimide (DTM) linkers (Behrens et al., 2015, Mol. Pharm. 12: 3986-3998), disulfidoaryl (TCEP) pyridazinedione-based linkers (Lee et al., 2016, Chem. Sci., 7: 799-802), or dibromopyridazinedione-based linkers (Maruani et al., 2015, Nat. Commun., 6: 6645).

[0446] In other embodiments, the targeting moiety T can be modified to include non-natural reactive groups (such as azide), which allow the complementary reactive groups on the joint to be conjugated to the joint. For example, the conjugation of the joint and the targeting moiety can utilize click chemistry (see, for example, Chio & Bane, 2020, Methods Mol.Biol., 2078: 83-97), such as azide-alkyne cycloaddition (AAC) reaction, which has been successfully used to develop antibody-drug conjugates. AAC reaction can be copper-catalyzed AAC (CuAAC) reaction, which relates to the coupling of azide and linear alkynes; or strain-promoted AAC (SPAAC) reaction, which relates to the conjugation of azide and cyclooctyne.

[0447] In some embodiments, the (e.g., cleavable) linker is substantially stable in the extratumoral environment (e.g., in the circulation). Such a stable linker may be characterized in that, after delivery to the tumor site and / or localization on the target cell surface and after a certain period of time, at least about 90%, about 80%, about 70%, about 60%, about 50%, or at least about 40% of the conjugate of Formula X is intact. The intact conjugate may, for example, have a DAR that is unchanged compared to the conjugate at the time of administration (e.g., within about ± 5%). In other words, in some embodiments, during the time that the conjugate resides in the extratumoral environment (e.g., in the systemic circulation and / or in non-target tissues or organs), the linker remains substantially uncleaved in such an environment. In some embodiments, such a linker may be cleaved in the extratumoral environment, but not to the extent that a useful dose of the intact conjugate is prevented from being delivered to the target cell (e.g., tumor cell). Whether the linker is substantially insensitive to the extratumoral environment can be determined by, for example, incubating the ISAC with plasma for a predetermined period of time (e.g., 2, 4, 8, 16, or 24 hours) and then quantifying the amount of free compound (e.g., those molecules cleaved from the conjugate) present in the plasma.

[0448] Various non-cleavable joints are known in the art for connecting the compounds of the present invention to the targeting moiety, and can be used in the conjugates of the present invention in certain embodiments. The example of a non-cleavable joint includes an N-succinimidyl ester or N-sulfosuccinimidyl ester moiety for reacting with a cell binding agent, and a joint based on a maleimido or haloacetyl moiety for reacting with a compound (e.g., according to any one of the compounds of formula (I)-(IV)), or vice versa. The example of such a non-cleavable joint is based on 4-[N-maleimidomethyl]cyclohexane-1-formyl sulfosuccinimidyl ester (sulfoSMCC). Sulfo-SMCC conjugation typically occurs via the maleimide group reacted with the sulfhydryl (thiol, -SH) on compound C, while sulfo-NHS esters are reactive to the primary amine on the targeting moiety T (as found in lysine and at the N-terminus of a protein or peptide). Other non-limiting examples of such linkers include those based on N-succinimidyl 4-(maleimidomethyl)cyclohexanecarboxylate (SMCC), 4-(N-maleimidomethyl)-cyclohexane-1-carboxy-(6-amidohexanoate) ("long chain" SMCC or LC-SMCC), N-succinimidyl maleimidoceanoate (KMUA), N-succinimidyl maleimidobutyrate (GMBS), maleimido Examples include iminohexanoic acid N-hydroxysuccinimide ester (EMCS), m-maleimidobenzoyl-N-hydroxysuccinimide ester (MBS), N-(maleimidoacetoxy)-succinimide ester (AMAS), 6-(maleimidopropionamido)hexanoic acid succinimide ester (SMPH), 4-(p-maleimidophenyl)-butyric acid N-succinimide ester (SMPB) and 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).

[0449] C. Preparation of Conjugates

[0450] The conjugates of formula (X) herein can be prepared by standard methods known in the art (see, e.g., 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, e.g., 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)). In addition, various antibody-drug conjugation services are commercially available 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).

[0451] In general, the preparation of the conjugates described herein comprises first preparing a compound-linker construct CL of formula (A) as described herein (which comprises one or more compounds of any one of formulas (I)-(IV) and a linker L), and then reacting the compound-linker construct (CL) with the compound-linker construct (CL). p A suitable group conjugated to the targeting moiety T. However, the attachment of a linker L to the targeting moiety T and the subsequent attachment of the targeting moiety-linker TL to one or more compounds described herein (e.g., those of Formula (I)) C remains an alternative approach that can be used in some embodiments. An exemplary approach is described in Example 4 herein.

[0452] In any of the above methods, suitable groups on the targeting moiety T for attachment to the linker L include sulfhydryl groups (e.g., on the side chain of a cysteine ​​residue), amino groups (e.g., on the side chain of a lysine residue), carboxylic acid groups (e.g., on the side chain of an aspartic acid or glutamic acid residue), and carbohydrate groups (e.g., on an Fc glycan portion). For example, the targeting moiety T may comprise one or more naturally occurring sulfhydryl groups that allow the targeting moiety T to be bonded to the linker L via the sulfur atom of the sulfhydryl group. In some embodiments, the targeting moiety T may comprise one or more lysine residues that can be chemically modified 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-pyridyldisulfide) propionate ("SPDP"), and 2-iminothiolane hydrochloride (Traut's reagent). In other embodiments, the targeting moiety T may comprise one or more carbohydrate groups that can be chemically modified to comprise one or more sulfhydryl groups.

[0453] In some embodiments, the carbohydrate group on the targeting moiety T can also be oxidized to provide an aldehyde (-CHO) group (see, e.g., Laguzza et al., 1989, J. Med. Chem. 32(3):548-55), which can then react with a linker L, e.g., via a hydrazine or hydroxylamine group on the linker L.

[0454] The targeting moiety T can also be modified to include 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 a reactive handle, 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 allow site-specific conjugation. In some embodiments, the targeting moiety T can be modified to include a non-natural reactive group (such as an azide) that allows conjugation to the linker via a complementary reactive group on the linker, for example, by click chemistry (see, e.g., Chio & Bane, 2020, Methods Mol. Biol., 2078:83-97).

[0455] Other protocols for modifying proteins (which the targeting moiety (T) herein may comprise or consist of) to allow attachment or association of a 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).

[0456] In those embodiments where the targeting moiety T is an antibody, several different reactive groups on the antibody can serve as sites for conjugation, including the epsilon-amino group on lysine residues, side chain carbohydrate moieties, side chain carboxylic acid groups on aspartic acid or glutamic acid residues, cysteine-cysteine ​​disulfide groups, and cysteine ​​thiol groups. The amino acids used for conjugation can be part of the native sequence of the antibody, or they can be introduced by site-specific engineering techniques known in the art as described herein.

[0457] Once conjugation is complete, the average number of compounds of Formula (I)-(IV) conjugated to the targeting moiety T (i.e., the "drug-antibody ratio" or DAR) can 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. In addition, the distribution of compound attachment patterns (e.g., the fraction of targeting moieties T containing 0, 1, 2, 3, etc., compounds C of Formula (I)) can also be optionally analyzed. Various techniques for measuring DAR distribution are known in the art, including MS (with or without an accompanying chromatographic separation step), hydrophobic interaction chromatography, reversed-phase HPLC, or isoelectric focusing gel electrophoresis (IEF) (see, e.g., Wakankar et al., 2011, mAbs, 3: 161-172).

[0458] D. Properties of the Conjugate

[0459] As described herein, the drug-antibody ratio (DAR) of a conjugate as disclosed herein (or "p" in formula X if r=1, otherwise the product of q times r) refers to the ratio of the number of compounds C of formula (I)-(IV) conjugated to the targeting moiety T.

[0460] As indicated above and reflected by the parameters p and r in formula (X), the targeting moiety "T" may be conjugated to more than one compound "C" of formula (I). One skilled in the art will appreciate that while any particular targeting moiety T is conjugated to an integer number of compounds C, analysis of preparations of conjugates to determine the ratio of compound C to targeting moiety T may yield non-integer results, reflecting statistical averages. This ratio of compound C to targeting moiety T may generally be referred to as the drug to antibody ratio or "DAR." Thus, preparations of conjugates having non-integer DARs are intended to be encompassed by formula (X). One skilled in the art will appreciate that the term "DAR" may be used to define conjugates comprising a targeting moiety other than an antibody.

[0461] In some embodiments, the DAR of the conjugate of Formula (X) herein is obtained by any combination of the linker to antibody ratio and / or the drug to linker ratio (e.g., where r is the ratio of compound-linker construct to antibody T, and p is the number of compounds C per linker L). For example, where r=2 and p=2, the number of compounds C of Formulae (I)-(IV) in the corresponding antibody-drug conjugate is 4 (e.g., each tumor-targeting antibody T is coupled to 2 compound-linker constructs, wherein each linker construct comprises 2 immunostimulatory compounds of Formulae (I)-(IV)).

[0462] 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.

[0463] In some embodiments, the DAR of the conjugates of formula (X) herein (i.e., the product of rx p in formula (X)) can have any value from about 1 to about 8, and thus can 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, 7.4, 7.5, 7.6, 7.8, 7.9, 8.0, 8.1, 8. , 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 conjugates of formula (X) herein may have a DAR of about 2 to about 8, about 2 to about 6, about 2 to about 5, about 3 to about 5, or about 2 to about 4.

[0464] As further described elsewhere herein, all compounds of the conjugate of formula (X) (eg, compounds of formula (I)-(IV)) may be identical. In other embodiments, two or more compounds of the conjugate of formula (X) may be different.

[0465] In some embodiments, the conjugates of Formula (X) herein can have an EC for agonizing TLR7 of about 10 nM to about 50 pM, about 5 nM to about 50 pM, about 1 nM to about 50 pM, about 750 pM to about 50 pM, about 600 pM to about 50 pM, about 500 pM to about 50 pM, about 400 pM to about 50 pM, about 300 pM to about 50 pM, about 200 pM to about 50 pM, or about 100 pM to about 50 pM. 50 value.

[0466] Without being bound by any theory, it is hypothesized that the anti-tumor activity of the conjugates of formula (X) herein can be mediated by innate immune cells. The conjugate, particularly in embodiments where the targeting moiety is an antibody or fragment thereof, can initially bind to tumor cells via the target TAA and bind to immune cells via FcγR engagement. Subsequently, the conjugate can be catabolized in the phagolysosome to release the TLR7 agonist, such as a compound of any one of formulas (I)-(IV). Stimulation of TLR7 in the phagolysosome can induce cytokine expression, thereby driving an anti-tumor response. Therefore, TLR7 agonism mediated by the compounds described herein can drive anti-tumor immunity in a subject.

[0467] In various embodiments, and as further described herein, the conjugate of formula (X) of the present disclosure comprising one or more immunomodulatory compounds can induce immune cells to produce and / or release one or more cytokines. Such immune cells can be human or mouse immune cells, for example, PBMC or splenocytes, and can be part of a cell population that further includes other cell types (such as other immune cells, tumor cells, etc.). Such induction of proinflammatory cytokines, for example, can be the result of the interaction between the immunostimulatory compound of the conjugate and the TLR7 of the immune cell.

[0468] In various embodiments, the conjugates 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 conjugates of formula (X) herein can induce the production of IL6. The induction of IL6 can include human IL6, mouse IL6, or both.

[0469] In various embodiments, the conjugates of Formula (X) herein may have an EC50 of <1 nM, <500 pM, <250 pM, <100 pM, or <50 pM or less for inducing cytokine production in human or murine immune cell populations. 50 In some embodiments, the conjugates of formula (X) herein may have an EC for inducing cytokine production in human or murine immune cell populations of <1 nM, <500 nM, or <250 nM. 50 In some embodiments, the conjugates of formula (X) herein may have an EC for inducing cytokine production in a human or murine immune cell population of about 1 nM to about 100 pM, about 750 pM to about 50 pM, about 500 pM to about 50 pM, about 250 pM to about 50 pM, or about 150 pM to about 50 pM. 50 Value. In various embodiments, the cytokine induced by the conjugate is IL6. In some embodiments, the induction of cytokine production by the conjugate can be determined via a human and / or murine immune cell assay, which can include incubating the conjugate with a cell population comprising immune cells (e.g., immune cells expressing TLR7) and tumor cells (e.g., tumor cells expressing an antigen to which the targeting moiety binds), followed by measuring the amount of the cytokine in the supernatant after a certain period of time, as further described herein, e.g., in Example 5.

[0470] In various embodiments, the conjugate of formula (X) comprising a compound of any one of formulas (I)-(IV) herein can elicit an immune response in vivo. In various embodiments, when administered to a subject (e.g., a human, non-human primate, or rodent) suffering from cancer, such an immune response can result in anti-tumor activity. Such anti-tumor activity can be measured by, for example, a reduction in tumor growth rate. In some embodiments, when administered to a group of test subjects bearing tumors, the anti-tumor activity of the conjugate of formula (X) herein is sufficient to reduce the tumor volume of one or more test subjects in the test subjects. In some embodiments, the conjugate of formula (X) herein can reduce the tumor volume of at least one test subject in the test subject by at least about 30%, 40%, 50%, 60%, 70%, 80%, or 90%.

[0471] In certain embodiments, the conjugates of formula (X) herein can show good in vivo tolerance. In some embodiments, the conjugates of formula (X) herein can be well tolerated in vivo at a dosage of at least about 1 mg / kg, at least about 3 mg / kg, at least about 15 mg / kg or about 45 mg / kg. When the total weight loss of the test subject at the end of the study period is no more than 20% relative to the baseline weight at the beginning of the study (i.e. before treatment), the conjugates herein are generally well tolerated. For example, the study period can be between 10 and 16 days. In some embodiments, the conjugates of formula (X) are well tolerated at a dosage that effectively reduces the tumor volume of the test subject.

[0472] Pharmaceutical composition

[0473] Compounds of formula (I)-(IV) and conjugates comprising compounds of formula (I)-(IV) can be formulated for therapeutic use. In certain embodiments, the disclosure further discloses a pharmaceutical composition comprising a compound of the disclosure (e.g., a compound according to any one of formulas (I)-(IV) (see, e.g., above "Compounds")) or a pharmaceutically acceptable salt thereof. Further disclosed herein is a pharmaceutical composition comprising a conjugate of formula (X) (see, e.g., above "Conjugates"). Any such pharmaceutical composition may further comprise a pharmaceutically acceptable carrier, diluent, or excipient. In some embodiments, the pharmaceutical composition is a therapeutic composition for treating a disease (e.g., cancer) in a subject in need thereof. Such pharmaceutical compositions can be prepared using well-known and readily available ingredients by known procedures.

[0474] Pharmaceutical compositions as described herein can be formulated for administration by, for example, oral (including, for example, buccal or sublingual), external, parenteral, rectal or vaginal routes, or by inhalation or spraying to a subject. The term parenteral as used herein includes subcutaneous injection and intradermal, intraarticular, intravenous, intramuscular, intravascular, intrasternal, intrathecal injection or infusion. Pharmaceutical compositions can be formulated into a form suitable for administration to a subject, for example, in the form of a syrup, elixir, tablet, lozenge, pastille, hard or soft capsule, pill, suppository, oily or aqueous suspension, dispersible powder or granules, emulsion, injection or solution. Pharmaceutical compositions can be provided as unit dose formulations.

[0475] Pharmaceutical compositions intended for oral use can be prepared as solid or fluid unit dosage forms. Fluid unit dosage forms can be prepared according to procedures known in the art for manufacturing pharmaceutical compositions, and such compositions can contain one or more agents, such as sweeteners, flavorings, coloring agents, and preservatives, to provide pharmaceutically aesthetic and palatable preparations. Elixirs can be prepared using a hydroalcoholic (e.g., ethanol) vehicle with a suitable sweetener such as sugar and / or saccharin and aromatic flavorings. Suspensions can be prepared with aqueous vehicles and suspending agents such as gum arabic, gum tragacanth, methylcellulose, etc.

[0476] Solid formulations, such as tablets, contain active ingredients (e.g., compounds and / or conjugates of the present disclosure) mixed with non-toxic, pharmaceutically acceptable excipients suitable for the manufacture of tablets. These excipients can be, for example, inert diluents such as calcium carbonate, sodium carbonate, lactose, calcium phosphate, or sodium phosphate; granulating agents and disintegrants 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 dicalcium phosphate, magnesium aluminum silicate, calcium sulfate, starch, lactose, methylcellulose, and functionally similar substances. Tablets can be uncoated, or they can be coated by known techniques, for example, to delay disintegration and absorption in the gastrointestinal tract, thereby providing a sustained action over a longer period of time. For example, a time-delaying agent such as glyceryl monostearate or glyceryl distearate can be employed.

[0477] Formulations for oral use may also be presented as hard gelatin capsules in which the active ingredient (e.g., a compound and / or conjugate of the present disclosure) is mixed with an inert solid diluent (e.g., calcium carbonate, calcium phosphate, or kaolin), or as soft gelatin capsules in which the active ingredient is mixed with water or an oil medium (e.g., peanut oil, liquid paraffin, or olive oil). Soft gelatin capsules are typically prepared by mechanically encapsulating a slurry of the active ingredient with an acceptable vegetable oil, light liquid petrolatum, or other inert oil.

[0478] Aqueous suspensions can contain active ingredients (such as compounds of the present disclosure and / or conjugates) mixed with excipients suitable for making aqueous suspensions. Such excipients include suspending agents, such as sodium carboxymethylcellulose, methylcellulose, hydroxypropyl methylcellulose, sodium alginate, polyvinyl pyrrolidone, tragacanth gum and gum arabic, and dispersants or wetting agents. Dispersants and wetting agents include, for example, naturally occurring phosphatides (such as lecithin), condensation products of alkylene oxide and fatty acids (such as polyoxyethylene stearate), condensation products of ethylene oxide and long-chain aliphatic alcohols (such as heptadecanol), condensation products of ethylene oxide and the partial ester derived from fatty acid and hexitol (such as polyoxyethylene sorbitol monooleate), or condensation products of ethylene oxide and the partial ester derived from fatty acid and hexitol anhydride (such as polyethylene sorbitan monooleate). The aqueous suspensions may also contain one or more preservatives (for example, ethyl or n-propyl p-hydroxybenzoate), one or more coloring agents, one or more flavoring agents, and / or one or more sweetening agents (for example, sucrose or saccharin).

[0479] Oily suspensions can be prepared by suspending the active ingredient (e.g., a compound and / or conjugate of the present disclosure) in a vegetable oil (e.g., peanut oil, olive oil, sesame oil, or coconut oil) or a mineral oil (such as liquid paraffin). Oily suspensions may contain thickeners, such as beeswax, hard paraffin, or cetyl alcohol. Sweeteners and flavorings, as described above, may be added to provide a palatable oral preparation. The suspension may optionally be preserved by the addition of an antioxidant, such as ascorbic acid.

[0480] Dispersible powders and granules suitable for preparing aqueous suspensions by adding water generally provide the active ingredient (e.g., a compound and / or conjugate of the present disclosure) in admixture with a dispersant or wetting agent, a suspending agent, and one or more preservatives. Suitable dispersants or wetting agents and suspending agents are exemplified by those mentioned above. One or more additional excipients, such as sweeteners, flavorings, and / or coloring agents, may also be present.

[0481] Pharmaceutical composition can also be in the form of oil-in-water emulsion.The oil phase can 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.The suitable emulsifier included in the oil-in-water emulsion includes, for example, naturally occurring gums (such as gum arabic or gum tragacanth), naturally occurring phosphatides (such as soybean, lecithin) or esters derived from fatty acids and hexitol anhydrides or partial esters (such as sorbitan monooleate) or the condensation products of such partial esters and ethylene oxide (such as polyoxyethylene sorbitan monooleate).Emulsion can also optionally contain sweeteners and / or flavorings.

[0482] The pharmaceutical compositions of the present disclosure can be in the form of sterile injectable aqueous or oily solutions or suspensions. Such suspensions can be prepared using suitable dispersants or wetting agents and suspending agents such as those mentioned above. Sterile injectable solutions or suspensions can contain active ingredients (e.g., compounds of the present disclosure and / or conjugates) in a non-toxic parenteral acceptable carrier or diluent. Acceptable carriers and diluents that can be used include, for example, 1,3-butanediol, water, Ringer's solution, or isotonic sodium chloride solution. In addition, sterile fixed oils can be used as carriers. For this purpose, various mild fixed oils can be used, including synthetic monoglycerides or diglycerides. In addition, fatty acids such as oleic acid are used in injectable formulations. Adjuvants, such as local anesthetics, preservatives, and / or buffers can also be included in injectable solutions or suspensions.

[0483] Pharmaceutical compositions can also be formulated as suppositories for rectal administration. These compositions can be prepared by mixing the active ingredient (e.g., a compound and / or conjugate of the present disclosure) with a suitable non-irritating excipient that is solid at room temperature but liquid at physiological temperature and will therefore melt in the rectum to release the drug. Such substances include cocoa butter and polyethylene glycol.

[0484] 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).

[0485] Medication boxes

[0486] In certain embodiments, a pharmaceutical composition comprising a compound of any one of Formulas (I)-(IV) or a conjugate of Formula (X) can be provided as part of a pharmaceutical kit or pack. The individual components of the kit can typically be packaged in separate containers. Depending on the formulation of the pharmaceutical composition, suitable containers include, for example, bottles, blister packs, intravenous solution bags, vials, and the like. In certain embodiments, the container can be in a form that allows administration to a subject, for example, an inhaler, a syringe, a pipette, an eye dropper, a pre-soaked gauze or pad, or other such device, from which the contents can be administered to a subject.

[0487] The kit may further include a label or package insert on or associated with one or more containers. The term "package insert" is used to refer to instructions customarily included in commercial packages of therapeutic products, which contain information about the indications, usage, dosage, administration, contraindications, and / or warnings regarding the use of such therapeutic products. The label or package insert may further include a notice in a form prescribed by a governmental agency regulating the manufacture, use, or sale of drugs or biological products, which notice reflects the agency's approval of the manufacture, use, or sale for human or animal administration. The label or package insert typically indicates that the compound or conjugate is used to treat a selected condition, such as cancer.

[0488] Where appropriate, one or more components of the kit may be lyophilized or provided in dry form (such as a powder or granules), and the kit may additionally contain a suitable solvent for reconstitution of one or more of the lyophilized or dried components.

[0489] How to use

[0490] Certain embodiments of the present disclosure relate to therapeutic uses of the purine derivative compounds described herein (e.g., compounds having a structure according to any one of Formulas (I)-(IV)) and conjugates comprising these compounds (e.g., conjugates of Formula (X)). Some embodiments of the present disclosure relate to the use of compounds according to any one of Formulas (I)-(IV) and / or conjugates of Formula (X) as therapeutic agents.

[0491] The TLR7-agonist compounds of the present disclosure (e.g., compounds according to any one of formulas (I)-(IV)) can exhibit cytotoxic activity against cancer cells, and the compounds of any one of formulas (I)-(IV) and conjugates comprising these compounds (i.e., conjugates of formula (X)) can therefore be used to inhibit the abnormal growth of cancer cells or tumor cells, inhibit the proliferation of cancer cells or tumor cells, and / or treat cancer in patients. In certain embodiments, the compounds of any one of formulas (I)-(IV) and the conjugates of formula (X) can be used to treat cancer in subjects in need. Therefore, some embodiments of the present disclosure relate to the use of compounds of any one of formulas (I)-(IV) and conjugates of general formula (X) as anticancer agents.

[0492] Certain embodiments of the present disclosure relate to methods of inhibiting the proliferation of cancer cells or tumor cells, comprising contacting the cells with a compound of any one of Formulas (I)-(IV) or a conjugate of Formula (X). Some embodiments relate to methods of killing cancer cells or tumor cells, comprising contacting the cells with a compound of any one of Formulas (I)-(IV) or a conjugate of Formula (X).

[0493] Various embodiments of the present disclosure relate to methods of exciting TLR7 using a compound of any one of Formulas (I)-(IV) or a conjugate of Formula (X) as described herein. In some embodiments, disclosed herein are methods of exciting TLR7 in vitro, comprising contacting a cell expressing TLR7 with a compound of any one of Formulas (I)-(IV) or a conjugate of Formula (X). In other embodiments, disclosed herein are methods of exciting TLR7 in a subject, comprising administering to the subject a compound of any one of Formulas (I)-(IV), a conjugate of Formula (X), or a pharmaceutical composition comprising such a compound or conjugate. 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 conjugate of Formula (X).

[0494] Using a compound or conjugate of the present disclosure (e.g., a compound of any one of Formulas (I)-(IV) or a conjugate of Formula (X)) to excite TLR7 on a cell in vitro or in vivo can trigger an immune response in the cell or subject. In some embodiments, disclosed herein is a method for eliciting an immune response in vitro, comprising contacting a cell with a compound of any one of Formulas (I)-(IV) or a conjugate of Formula (X). The cell can be a mammalian cell. The mammalian cell can be an immune cell, such as a lymphocyte (e.g., a T cell) or a phagocyte (e.g., a neutrophil, a macrophage, a dendritic cell, an eosinophil, or a monocyte). In yet other embodiments, disclosed herein is a method for eliciting an immune response in a subject in need thereof, comprising administering a compound of any one of Formulas (I)-(IV) or a conjugate of Formula (X) to the subject. This method may further comprise exciting TLR7 in the subject by administering a compound of any one of Formulas (I)-(IV) or a conjugate of Formula (X), thereby triggering an immune response in the subject. In various embodiments, disclosed herein is a method for eliciting a local immune response in a subject, the method comprising administering to a subject in need thereof a conjugate of formula (X) herein, the conjugate comprising a compound of any one of one or more formulas (I)-(IV). After administering (e.g., parenteral administration) the conjugate of formula (X) to the subject, the method may further comprise accumulating the conjugate at a target site (e.g., a target organ or tissue) in the subject, and eliciting a local immune response at the target site. As used herein, the term "local immune response" is relative to a systemic immune response and generally refers to an immune response that occurs at a specific location (e.g., an organ) in a subject or within a certain tissue type present at one or more locations. For example, a local anti-tumor immune response is generally understood herein to refer to an immune response that occurs at a tumor site or multiple tumor sites (e.g., in the case of metastasis) comprising tumor tissue. Local immune responses can be measured or detected by changes in one or more physiological parameters (such as local concentrations of biomarkers, such as cytokines, small molecules, co-stimulatory molecules, and / or production / secretion of factors involved in the inflammatory cascade or regulation), and / or changes in immune cell populations.

[0495] In some embodiments, the method of exciting TLR7 using a compound of any one of Formulas (I)-(IV) or a conjugate of Formula (X) can further include activating the TLR7 signaling pathway. Such activation can result in a measurable change (i.e., an increase or decrease) in the expression of one or more intermediates in the TLR7 signaling pathway. Intermediates of 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, for example, 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 conjugates of the present disclosure include, but are not limited to, tumor necrosis factor (TNF; also known as TNFα or cachectin), interleukin (IL)-1α, IL1β, IL2; IL5, IL6, IL8, IL15, IL18, interferon gamma (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 kinases; monocyte chemoattractant proteins 1 and 2 (MCP-1, MCP-2); macrophage migration inhibitory factor (MIF) and high mobility group box protein 1 (HMGB-1).

[0496] Methods for assessing 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. Biomolecule detection can be performed directly on a sample collected from a subject, or the sample can be processed between sample collection and analysis.

[0497] Some embodiments of the present disclosure relate to methods of treating a subject with cancer by administering to the subject a compound of formula (I) or a conjugate of formula (X) or a pharmaceutical composition comprising such a compound or conjugate. In this context, treatment with a compound of any one of formulas (I)-(IV) or a conjugate of formula (X) can result in one or more of the following: a reduction in tumor size, a slowing down or prevention of an increase in tumor size, a prolonged disease-free survival time between the disappearance or removal of a tumor and its reappearance, prevention of subsequent occurrence of a tumor (e.g., metastasis), a prolonged time to progression, a reduction in one or more adverse symptoms associated with the tumor, and / or a prolonged overall survival time of the subject with cancer.

[0498] Certain embodiments relate to the use of a compound of any one of formula (I)-(IV) or a conjugate 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 formula (I)-(IV) or a conjugate of formula (X) in a method of inhibiting cancer cell proliferation and / or killing cancer cells in vitro. Some embodiments relate to the use of a compound of any one of formula (I)-(IV) or a conjugate of formula (X) in a method of inhibiting cancer cell proliferation and / or killing cancer cells in vivo in a subject suffering from cancer.

[0499] Examples of cancers that can be treated using the compounds or conjugates disclosed herein in certain embodiments of the present disclosure include blood tumors, including leukemias, myelomas, and lymphomas; carcinomas, including adenocarcinomas and squamous cell carcinomas; melanomas and sarcomas. Carcinomas and sarcomas are often also referred to as "solid tumors." Examples of common solid tumors that can 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, gastric cancer, uterine cancer, non-small cell lung cancer (NSCLC), and colorectal cancer. Various forms of lymphoma can also lead to the formation of solid tumors and, therefore, can also be considered solid tumors in certain cases.

[0500] In other embodiments, the compound of any one of Formulas (I)-(IV) herein or the conjugate of Formula (X) can be used as a vaccine adjuvant. In yet other embodiments, the compound of any one of Formulas (I)-(IV) herein or the conjugate of Formula (X) can be used in a method of treating a viral infection in a subject in need thereof. Such a method may comprise administering an effective amount of the compound or conjugate to the subject, thereby treating the viral infection in the subject.

[0501] Certain embodiments of the present disclosure

[0502] In certain embodiments, the present disclosure relates to any one or more of embodiments 1-140.

[0503] Embodiment 1. A compound of formula (I):

[0504]

[0505] its tautomers, protected forms and / or pharmaceutically acceptable salts,

[0506] in:

[0507] X is O or NH;

[0508] Y is N or CH;

[0509] R 1 is an optionally substituted C2-C6 alkyl group or a branched and optionally substituted C3-C8 hydroxyalkyl group;

[0510] R 2 is H, halogen, optionally substituted C1-C6 alkyl, or optionally substituted C1-C6 alkoxy;

[0511] R 3 and R 4 H, QR independently 5 , or R 3 and R 4 Together with N, it forms an optionally substituted C2-C6-heterocycloalkyl ring, an optionally substituted C3-C7-heterobicycloalkyl ring, an optionally substituted C6-C 10 - a heterospirocycloalkyl ring or an optionally substituted heteroaryl ring;

[0512] Q is a bond, optionally substituted C1-C6-alkyl, optionally substituted C2-C6-alkenyl or Where * is the same as R 5 's attachment point, and # is the attachment point with N;

[0513] R 5 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;

[0514] 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, it forms an optionally substituted C2-C6-heterocycloalkyl ring, an optionally substituted C3-C7-heterobicycloalkyl ring, an optionally substituted C6-C 10 - a heterospirocycloalkyl ring or an optionally substituted heteroaryl ring;

[0515] 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, optionally substituted aryl or optionally substituted heteroaryl;

[0516] n is an integer from 1 to 4;

[0517] m is an integer from 0 to 4; and

[0518] a and b are each independently an integer from 0 to 4,

[0519] The condition is that when R 2 When it is H, then:

[0520] (i)R 1 is a branched and optionally substituted C3-C8 hydroxyalkyl group;

[0521] (ii) X is NH and R 1 is an optionally substituted C5-C6 alkyl group; or

[0522] (iii) m is 0 or 1, and R 3 and R 4 Together with N, it forms an unsubstituted piperazinyl ring or an optionally substituted C6-C 10 -heterospirocycloalkyl ring.

[0523] The compound as described in embodiment 1, wherein R 2 is halogen, optionally substituted C1-C6 alkyl, or optionally substituted C1-C6 alkoxy.

[0524] Embodiment 2. The compound according to embodiment 1, wherein R 2 is halogen, optionally substituted C1-C6 alkyl, or optionally substituted C1-C6 alkoxy.

[0525] Embodiment 3. A compound as described in any one of embodiments 1-2, wherein R 2 is halogen or optionally substituted C1-C6 alkoxy.

[0526] Embodiment 4. A compound as described in any one of embodiments 1-3, wherein R 2 is halogen or unsubstituted C1-C6 alkoxy.

[0527] Embodiment 5. A compound as described in any one of embodiments 1-4, wherein R 2 is fluorine or methoxy.

[0528] Embodiment 6. The compound of any one of embodiments 1-5, wherein X is O.

[0529] Embodiment 7. The compound of any one of embodiments 1-5, wherein X is NH.

[0530] Embodiment 8. A compound as described in any one of embodiments 1-7, wherein R 1 is an optionally substituted C2-C6 alkyl group.

[0531] Embodiment 9. A compound as described in any one of embodiments 1-7, wherein R 1is a branched and optionally substituted C3-C8 hydroxyalkyl group.

[0532] Embodiment 10. A compound as described in any one of embodiments 1-9, wherein R 3 and R 4 are independently H or QR 5 .

[0533] Embodiment 11. A compound as described in any one of embodiments 1-10, wherein R 3 and R 4 Both are H or R 3 and R 4 All are QR 5 .

[0534] Embodiment 12. A compound as described in any one of embodiments 1-10, wherein R 3 H and R 4 QR 5 .

[0535] Embodiment 13. The compound of embodiment Error! Referene source not found., wherein 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, optionally substituted aryl or optionally substituted heteroaryl.

[0536] Embodiment 14. The compound of embodiment Error! Reference source not found., wherein Q is an optionally substituted C1-C6-alkyl group and R 5 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.

[0537] Embodiment 15. The compound of embodiment Error! Referene source not found., wherein Q is an optionally substituted C2-C6-alkenyl and R 5 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.

[0538] Embodiment 16. The compound of any one of Embodiments 1 to Error! Referene source not found., wherein Q is:

[0539]

[0540] And among them:

[0541] 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, optionally substituted aryl, or optionally substituted heteroaryl; and

[0542] a and b are each independently an integer from 0 to 4.

[0543] Embodiment 17. The compound of embodiment 17, wherein Q is selected from the group consisting of:

[0544]

[0545] wherein a and b are each independently 1, 2 or 3.

[0546] Embodiment 18. A compound as described in any one of Embodiments 1-Error! Referene source not found. or Error! Referene source not found.-Error! Referene source not found., wherein R 5 NR 6 R 7 And R 6 and R 7 Both are H.

[0547] Embodiment 19. A compound as described in any one of Embodiments 1-Error! Referene source not found. or Error! Referene source not found.-Error! Referene source not found., wherein R 5 NR 6 R 7 , R 6 H and R 7 is an optionally substituted C1-C6 alkyl group.

[0548] Embodiment 20. A compound as described in any one of Embodiments 1-Error! Referene source not found. or Error! Referene source not found.-Error! Referene source not found., wherein R 5 NR 6 R 7 , and R 6 and R 7 Together with N, it forms an optionally substituted C2-C6-heterocycloalkyl ring, an optionally substituted C3-C7-heterobicycloalkyl ring, an optionally substituted C6-C 10 - a heterospirocycloalkyl ring or an optionally substituted heteroaryl ring.

[0549] Embodiment 21. A compound as described in any one of embodiments 1-9, wherein R 3 and R 4 Together with N, it forms an optionally substituted C2-C6-heterocycloalkyl ring, an optionally substituted C3-C7-heterobicycloalkyl ring, an optionally substituted C6-C 10 - a heterospirocycloalkyl ring or an optionally substituted heteroaryl ring.

[0550] Embodiment 22. The compound of any one of embodiments 1-21, wherein n is 1.

[0551] Embodiment 23. The compound of any one of embodiments 1-22, wherein m is an integer from 0 to 3 or from 1 to 3.

[0552] Embodiment 24. The compound according to embodiment 1, wherein R 2 H and R 1 is a branched and optionally substituted C3-C8 hydroxyalkyl group.

[0553] Embodiment 25. The compound according to embodiment 13, wherein R 1is a branched unsubstituted C3-C8 hydroxyalkyl group.

[0554] Embodiment 26. The compound according to embodiment 1, wherein R 2 is H, X is NH, and R 1 is an optionally substituted C5-C6 alkyl group.

[0555] Embodiment 27. The compound of embodiment Error! Referene source not found., wherein R 1 It is an unsubstituted C5-C6 alkyl group.

[0556] Embodiment 28. The compound according to embodiment 1, wherein R 2 is H, m is 0 or 1, and R 3 and R 4 Together with N, it forms an unsubstituted piperazinyl ring or an optionally substituted C6-C 10 -heterospirocycloalkyl ring.

[0557] Embodiment 29. The compound of embodiment Error! Referene source not found., wherein R 3 and R 4 Together with N, it forms an unsubstituted piperazinyl ring.

[0558] Embodiment 30. The compound of embodiment Error! Referene source not found., wherein R 3 and R 4 Together with N, it forms an optionally substituted C6-C 10 -heterospirocycloalkyl ring.

[0559] Embodiment 31. A compound as described in any one of the embodiments Error! Referene source not found.-Error! Referene source not found., wherein m is 0.

[0560] Embodiment 32. A compound as described in any one of Embodiments 1 to 3, wherein m is 1.

[0561] Embodiment 33. A compound as described in any one of Embodiments 1 to Error! Referene source not found., wherein each alkyl, alkenyl, cycloalkyl, spiroalkyl, heterocycloalkyl, heterospiroalkyl, aryl and heteroaryl are optionally replaced by -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)-spiroalkyl, (C0-C4 alkyl)-heterospiroalkyl, (C0-C4 alkyl)- wherein each of the alkyl, hydroxyalkyl, C1-C4 aminoalkyl, C1-C4 carboxyalkyl, cycloalkyl, heterocycloalkyl, spiroalkyl, heterospiroalkyl, aryl or heteroaryl groups may itself be substituted by 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.

[0562] Embodiment 34. The compound of Embodiment 34, wherein each alkyl, alkenyl, cycloalkyl, spirocycloalkyl, heterocycloalkyl, heterospirocycloalkyl, aryl, and heteroaryl is optionally substituted with one or more substituents selected from the group consisting of: -NH2, -C02H, -OH, carbonyl, halogen, unsubstituted C1-C4 alkyl, unsubstituted C1-C4 hydroxyalkyl, unsubstituted C1-C4 aminoalkyl, and unsubstituted C1-C4 carboxyalkyl.

[0563] Embodiment 35. The compound of Embodiment 1, wherein the compound is selected from any one of compounds 100-195 listed in Table 1.

[0564] Embodiment 36. The compound according to embodiment 1, which has the structure of formula (II):

[0565]

[0566] in:

[0567] X is O or NH;

[0568] Y is N or CH;

[0569] R 1 is an optionally substituted C2-C6 alkyl group or a branched and optionally substituted C3-C8 hydroxyalkyl group;

[0570] R3 and R 4 H, QR independently 5 , or R 3 and R 4 Together with N, it forms an optionally substituted C2-C6-heterocycloalkyl ring, an optionally substituted C3-C7-heterobicycloalkyl ring, an optionally substituted C6-C 10 - a heterospirocycloalkyl ring or an optionally substituted heteroaryl ring;

[0571] Q is a bond, optionally substituted C1-C6-alkyl, optionally substituted C2-C6-alkenyl or Where * is the same as R 5 's attachment point, and # is the attachment point with N;

[0572] R 5 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;

[0573] 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, it forms an optionally substituted C2-C6-heterocycloalkyl ring, an optionally substituted C3-C7-heterobicycloalkyl ring, an optionally substituted C6-C 10 - a heterospirocycloalkyl ring or an optionally substituted heteroaryl ring;

[0574] 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, optionally substituted aryl or optionally substituted heteroaryl;

[0575] m is an integer from 0 to 4; and

[0576] a and b are independently integers from 0 to 4.

[0577] Embodiment 37. The compound of embodiment 15, wherein X is O and R 1 is an optionally substituted C2-C6 alkyl group.

[0578] Embodiment 38. A compound as described in any one of Embodiments 15-Error! Referene source not found., wherein R 1 It is an unsubstituted C2-C6 alkyl group.

[0579] Embodiment 39. The compound of any one of Embodiments 15-Error! Referene source not found., wherein Y is CH.

[0580] Embodiment 40. The compound of any one of Embodiments 15-Error! Referene source not found., wherein m is 0 or 1, 1 or 2, or 1 or 3.

[0581] Embodiment 41. A compound as described in any one of Embodiments 15-Error! Referene source not found., wherein R 3 and R 4 Both are H.

[0582] Embodiment 42. A compound as described in any one of Embodiments 15-Error! Referene source not found., wherein R 3 and R 4 Together with N, it forms an optionally substituted C2-C6-heterocycloalkyl ring, an optionally substituted C3-C7-heterobicycloalkyl ring, an optionally substituted C6-C 10 - a heterospirocycloalkyl ring or an optionally substituted heteroaryl ring.

[0583] Embodiment 43. A compound as described in any one of Embodiments 15-Error! Referene source not found., wherein R 3 and R 4 Both are H or QR 5 , or R 3 H and R 4 QR 5 .

[0584] Embodiment 44. The compound of embodiment Error! Referene source not found., wherein 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, optionally substituted aryl or optionally substituted heteroaryl.

[0585] Embodiment 45. The compound of embodiment Error! Reference source not found., wherein Q is an optionally substituted C1-C6-alkyl group and R 5 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.

[0586] Embodiment 46. The compound of embodiment Error! Referene source not found., wherein Q is an optionally substituted C2-C6-alkenyl and R 5 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.

[0587] Embodiment 47. A compound as described in any one of Embodiments 15-Error! Referene source not found. or Error! Referene source not found.-Error! Referene source not found., wherein R 5 NR 6 R 7 And R 6 and R 7 Both are H.

[0588] Embodiment 48. A compound as described in any one of Embodiments 15-Error! Referene source not found. or Error! Referene source not found.-Error! Referene source not found., wherein R 5 NR 6 R 7 , R 6 H and R7 is an optionally substituted C1-C6 alkyl group.

[0589] Embodiment 49. A compound as described in any one of Embodiments 15-Error! Referene source not found. or Error! Referene source not found.-Error! Referene source not found., wherein R 6 and R 7 Together with N, it forms an optionally substituted C2-C6-heterocycloalkyl ring, an optionally substituted C3-C7-heterobicycloalkyl ring, an optionally substituted C6-C 10 - a heterospirocycloalkyl ring or an optionally substituted heteroaryl ring.

[0590] Embodiment 50. The compound of any one of Embodiments 15-Error! Referene source not found., wherein m is 1 or 2, or 1 or 3.

[0591] Embodiment 51. The compound of Embodiment 36, wherein the compound is selected from Sub-Table 1A.

[0592] Embodiment 52. A compound as described in any one of Embodiments 1-9, Error! Referene source not found. or Error! Referene source not found., wherein R 3 and R 4 Together with N, forms a ring selected from the group consisting of:

[0593]

[0594] in:

[0595] Z is N or CH; and

[0596] R 8 is H, NH2, optionally substituted C1-C6-alkyl, optionally substituted C1-C6-heteroalkyl, optionally substituted C1-C6-aminoalkyl or optionally substituted C3-C6-heterocycloalkyl.

[0597] Embodiment 53. The compound of embodiment Error! Referene source not found., wherein R 3 and R 4 Together with N, forms a ring selected from the group consisting of:

[0598]

[0599] Embodiment 54. A compound as described in any of Embodiments 1-9 or 21, wherein the compound is selected from Sub-Table 1B.

[0600] Embodiment 55. A compound as described in any one of Embodiments 1-Error! Referene source not found., wherein R 3 and R 4 All are QR 5 .

[0601] Embodiment 56. The compound of embodiment 55, wherein the compound is:

[0602]

[0603]

[0604] Embodiment 57. The compound of embodiment 1-11, wherein R 3 and R 4 Both are H.

[0605] Embodiment 58. The compound of embodiment 57, wherein the compound is:

[0606]

[0607] Embodiment 59. The compound of Embodiment 13, wherein the compound is selected from Sub-Table 1C.

[0608] Embodiment 60. The compound of embodiment 15 or embodiment 46, wherein the compound is:

[0609]

[0610] Embodiment 61. A compound as described in any of Embodiments 1-10 or 14, wherein the compound is selected from Sub-Table 1D.

[0611] Embodiment 62. A compound as described in any one of embodiments 1-10 or 16-17, wherein R 6 H and R 7 is H, an optionally substituted C1-C6 alkyl group, or R 6 and R 7 Together with N, it forms an optionally substituted C2-C6-heterocycloalkyl ring, an optionally substituted C3-C7-heterobicycloalkyl ring, an optionally substituted C6-C 10 - a heterospirocycloalkyl ring or an optionally substituted heteroaryl ring.

[0612] Embodiment 63. A compound as described in Embodiment 62, wherein the compound is selected from Sub-Table 1E.

[0613] Embodiment 64. The compound of any one of embodiments 1-9 or 36, wherein the compound is:

[0614]

[0615] Embodiment 65. The compound of embodiment 1 or 13-Error! Referene source not found., wherein the compound is:

[0616]

[0617] Embodiment 66. A compound of formula (III):

[0618]

[0619] its tautomers, protected forms and / or pharmaceutically acceptable salts,

[0620] in:

[0621] R 1a is an optionally substituted C2-C6 alkyl group or a branched and optionally substituted C3-C8 hydroxyalkyl group;

[0622] R 2a is halogen, optionally substituted C1-C6 alkyl, or optionally substituted C1-C6 alkoxy;

[0623] R 3a and R 4a H, QR independently 5a , or R 3a and R 4a Together with N, it forms an optionally substituted C2-C6-heterocycloalkyl ring, an optionally substituted C3-C7-heterobicycloalkyl ring, an optionally substituted C6-C 10 - a heterospirocycloalkyl ring or an optionally substituted heteroaryl ring;

[0624] Q is a bond, optionally substituted C1-C6-alkyl, optionally substituted C2-C6-alkenyl or Where * is the same as R 5 's attachment point, and # is the attachment point with N;

[0625] R 5a 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;

[0626] R 6a and R 7a are independently H, optionally substituted C1-C6 alkyl, optionally substituted C1-C6 alkoxycarbonyl, or R 6a and R 7a Together with N, it forms an optionally substituted C2-C6-heterocycloalkyl ring, an optionally substituted C3-C7-heterobicycloalkyl ring, an optionally substituted C6-C 10 - a heterospirocycloalkyl ring or an optionally substituted heteroaryl ring;

[0627] W is an optionally substituted C3-C6-cycloalkyl, an optionally substituted C3-C6-heterocycloalkyl, an optionally substituted C5-C7-heterobicycloalkyl, an optionally substituted C6-C 10 -heterospirocycloalkyl, optionally substituted aryl or optionally substituted heteroaryl;

[0628] u is an integer from 1 to 3;

[0629] v is an integer from 0 to 4; and

[0630] a and b are independently integers from 0 to 4.

[0631] Embodiment 67. The compound according to embodiment 28, wherein R 1a is an optionally substituted C2-C6 alkyl group.

[0632] Embodiment 68. A compound as described in any one of Embodiments 28-Error! Referene source not found., wherein R 1a It is an unsubstituted C2-C6 alkyl group.

[0633] Embodiment 69. The compound of any one of Embodiments 28-Error! Referene source not found., wherein 2a is halogen or optionally substituted C1-C6 alkoxy.

[0634] Embodiment 70. The compound of any one of Embodiments 28-Error! Referene source not found., wherein R 2a is halogen or unsubstituted C1-C6 alkoxy.

[0635] Embodiment 71. A compound as described in any one of Embodiments 28-Error! Referene source not found., wherein R 2a is fluorine or methoxy.

[0636] Embodiment 72. The compound of any one of Embodiments 28-Error! Referene source not found., wherein R 3a and R 4a Together with N, it forms an optionally substituted C2-C6-heterocycloalkyl ring, an optionally substituted C3-C7-heterobicycloalkyl ring, an optionally substituted C6-C 10 - a heterospirocycloalkyl ring or an optionally substituted heteroaryl ring.

[0637] Embodiment 73. A compound as described in any one of Embodiments 28-Error! Referene source not found., wherein R 3a and R 4a Both are H.

[0638] Embodiment 74. A compound as described in any one of Embodiments 28-Error! Referene source not found., wherein R 3a and R 4a are independently H or QR 5 .

[0639] Embodiment 75. The compound of embodiment Error! Referene source not found., wherein R 3a H and R 4a QR 5a .

[0640] Embodiment 76. A compound as described in any one of the embodiments Error! Referene source not found.-Error! Referene source not found., wherein Q is a bond and R 5a 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, optionally substituted aryl or optionally substituted heteroaryl.

[0641] Embodiment 77. A compound as described in any one of the embodiments Error! Referene source not found.-Error! Referene source not found., wherein Q is an optionally substituted C1-C6-alkyl group and R 5a 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.

[0642] Embodiment 78. A compound as described in any one of the embodiments Error! Referene source not found.-Error! Referene source not found., wherein Q is an optionally substituted C2-C6-alkenyl and R 5a 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.

[0643] Embodiment 79. The compound of any one of Embodiments 28-Error! Referene source not found., wherein Q is:

[0644]

[0645] And among them:

[0646] 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, optionally substituted aryl, or optionally substituted heteroaryl; and

[0647] a and b are independently integers from 0 to 4.

[0648] Embodiment 80. The compound of embodiment 80, wherein Q is selected from the group consisting of:

[0649]

[0650] wherein a and b are independently 1, 2 or 3.

[0651] Embodiment 81. A compound as described in any one of Embodiments 28-Error! Referene source not found. or Error! Referene source not found.-Error! Referene source not found., wherein R 6a and R 7a Both are H.

[0652] Embodiment 82. A compound as described in any one of Embodiments 28-Error! Referene source not found. or Error! Referene source not found.-Error! Referene source not found., wherein R 6a H and R 7a is an optionally substituted C1-C6 alkyl group.

[0653] Embodiment 83. A compound as described in any one of Embodiments 28-Error! Referene source not found. or Error! Referene source not found.-Error! Referene source not found., wherein R 6a and R 7a Together with N, it forms an optionally substituted C2-C6-heterocycloalkyl ring, an optionally substituted C3-C7-heterobicycloalkyl ring, an optionally substituted C6-C 10 - a heterospirocycloalkyl ring or an optionally substituted heteroaryl ring.

[0654] Embodiment 84. The compound of any one of Embodiments 28-Error! Referene source not found., wherein u is 1.

[0655] Embodiment 85. The compound of any one of Embodiments 28-Error! Referene source not found., wherein v is an integer from 0 to 4, from 1 to 4, from 1 to 3, or from 1 to 2.

[0656] Embodiment 86. The compound of any one of Embodiments 28-Error! Referene source not found., wherein R 3a and R 4a Together with N, forms a ring selected from the group consisting of:

[0657]

[0658] in:

[0659] Z is N or CH; and

[0660] R 8a is H, NH2, optionally substituted C1-C6-alkyl, optionally substituted C1-C6-heteroalkyl, optionally substituted C1-C6-aminoalkyl or optionally substituted C3-C6-heterocycloalkyl.

[0661] Embodiment 87. The compound of embodiment Error! Referene source not found., wherein R 3a and R 4a Together with N, forms a ring selected from the group consisting of:

[0662]

[0663] Embodiment 88. The compound according to embodiment 66, which has the structure of formula (IV):

[0664]

[0665] in:

[0666] R 1a is an unsubstituted C2-C6 alkyl group or a branched and unsubstituted C3-C8 hydroxyalkyl group;

[0667] R 3a and R 4a H, QR independently 5a , or R 3a and R 4a Together with N, it forms an optionally substituted C2-C6-heterocycloalkyl ring, an optionally substituted C3-C7-heterobicycloalkyl ring, an optionally substituted C6-C 10 - a heterospirocycloalkyl ring or an optionally substituted heteroaryl ring;

[0668] Q is a bond, optionally substituted C1-C6-alkyl, optionally substituted C2-C6-alkenyl or Where * is the same as R 5 's attachment point, and # is the attachment point with N;

[0669] R 5a 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;

[0670] R 6a and R 7a are independently H, optionally substituted C1-C6 alkyl, optionally substituted C1-C6 alkoxycarbonyl, or R 6a and R 7a Together with N, it forms an optionally substituted C2-C6-heterocycloalkyl ring, an optionally substituted C3-C7-heterobicycloalkyl ring, an optionally substituted C6-C 10 - a heterospirocycloalkyl ring or an optionally substituted heteroaryl ring;

[0671] 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, optionally substituted aryl or optionally substituted heteroaryl;

[0672] v is an integer from 0 to 4; and

[0673] a and b are independently integers from 0 to 4.

[0674] Embodiment 89. A compound as described in embodiment 66 or embodiment 88, wherein each alkyl, alkenyl, cycloalkyl, spiroalkyl, heterocycloalkyl, heterospiroalkyl, aryl and heteroaryl are optionally replaced by -NH2, -C02H, -OH, carbonyl, halogen, C1-C4 alkyl, C1-C4 hydroxyalkyl, C1-C4 aminoalkyl, C1-C4 carboxylalkyl, (C0-C4 alkyl)-cycloalkyl, (C0-C4 alkyl)-heterocycloalkyl, (C0-C4 alkyl)-spiroalkyl, (C0-C4 alkyl)-heterospiroalkyl, ( and (C0-C4 alkyl)-heteroaryl, and wherein each of the alkyl, hydroxyalkyl, C1-C4 aminoalkyl, C1-C4 carboxyalkyl, cycloalkyl, heterocycloalkyl, spiroalkyl, heterospiroalkyl, aryl or heteroaryl groups may itself be substituted by 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.

[0675] Embodiment 90. The compound of embodiment 30, wherein each alkyl, alkenyl, cycloalkyl, spiroalkyl, heterocycloalkyl, heterospiroalkyl, aryl, and heteroaryl is optionally substituted with one or more substituents selected from the group consisting of: -NH2, -C02H, -OH, carbonyl, halogen, unsubstituted C1-C4 alkyl, unsubstituted C1-C4 hydroxyalkyl, unsubstituted C1-C4 aminoalkyl, and unsubstituted C1-C4 carboxyalkyl.

[0676] Embodiment 91. A compound as described in Embodiment 66 or Embodiment 88, wherein the compound is selected from SubTable 1A.

[0677] Embodiment 92. The compound of embodiment 66 or embodiment 88, wherein the compound is:

[0678]

[0679] Embodiment 93. A compound as described in any of embodiments 1-92, wherein the compound has an EC for agonizing human or murine TLR7 of <500 nM, <250 nM, or <100 nM as determined in a reporter gene assay. 50 values, and / or an EC50 of <5 μM, <1 μM, <500 nM or <100 nM to induce IL6 production in a human or murine immune cell population as determined in an immune cell assay 50 value.

[0680] Embodiment 94. A pharmaceutical composition comprising a compound according to any one of embodiments 1-92 and a pharmaceutically acceptable carrier or diluent.

[0681] Embodiment 95. A method of agonizing TLR7, the method comprising contacting a cell expressing TLR7 with a compound of any one of embodiments 1-33, thereby agonizing TLR7.

[0682] Embodiment 96. A method of inducing cytokine release from a cell, the method comprising contacting the cell with a compound of any one of embodiments 1-33, thereby inducing the release of the cytokine from the cell.

[0683] Embodiment 97. A method of stimulating an immune response in a subject in need thereof, comprising administering to the subject an effective amount of a compound of any one of embodiments 1-33.

[0684] Embodiment 98. The method of embodiment 98, wherein the compound agonizes TLR7 in the subject, thereby stimulating the immune response in the subject.

[0685] Embodiment 99. A method of inhibiting the proliferation of cancer cells, the method comprising contacting a cell population comprising the cancer cells with an effective amount of a compound of any one of embodiments 1-33.

[0686] Embodiment 100. A method of killing cancer cells, comprising contacting a cell population comprising the cancer cells with an effective amount of a compound of any one of Embodiments 1-33.

[0687] Embodiment 101. The method of any one of Embodiments 101-10, wherein the cell population comprises immune cells.

[0688] Embodiment 102. A method of treating cancer in a subject in need thereof, comprising administering to the subject an effective amount of a compound of any one of Embodiments 1-33.

[0689] Embodiment 103. The method of embodiment 103, wherein the compound agonizes TLR7 in the subject, thereby treating the cancer in the subject.

[0690] Embodiment 104. A compound according to any one of embodiments 1-92 for use in therapy.

[0691] Embodiment 105. A compound according to any one of embodiments 1-92 for use in the treatment of cancer.

[0692] Embodiment 106. Use of a compound according to any one of Embodiments 1-92 in the manufacture of a medicament for the treatment of cancer.

[0693] Embodiment 107. A compound-linker construct having formula (A):

[0694] (A)

[0695] L-(C) p

[0696] in:

[0697] L is the connector;

[0698] C is a compound of any one of embodiments 1-92; and

[0699] p is an integer from 1 to 5.

[0700] Embodiment 108. The compound-linker construct of embodiment 107, wherein L is a cleavable linker.

[0701] Embodiment 109. The compound-linker construct of embodiment 107, wherein L is a non-cleavable linker.

[0702] Embodiment 110. The compound-linker construct of any one of embodiments 107-108, wherein L comprises a dipeptide, a tripeptide, a tetrapeptide, or a combination thereof.

[0703] Embodiment 111. The compound-linker construct of any one of embodiments 107-110, wherein L comprises a polyethylene glycol (PEG) moiety.

[0704] Embodiment 112. The compound-linker construct of any one of embodiments 107-108, wherein L is a protease-cleavable linker.

[0705] Embodiment 113. The compound-linker construct of any one of embodiments 107-112, wherein C is the compound of embodiment 36.

[0706] Embodiment 114. The compound-linker construct of any one of embodiments 107-112, wherein C is the compound of embodiment 66.

[0707] Embodiment 115. The compound-linker construct of embodiment 107, wherein the compound-linker is selected from Table 2B.

[0708] Embodiment 116. The compound-linker construct of any one of embodiments 107-115, wherein p is 1, 2, or 3.

[0709] Embodiment 117. A conjugate having formula (X):

[0710] (X)

[0711] T-[L-(C) p ] r

[0712] in:

[0713] T is the targeting moiety;

[0714] L is the connector;

[0715] C is a compound according to any one of embodiments 1-33;

[0716] p is an integer from 1 to 5; and

[0717] The value of r is from about 1 to about 8.

[0718] Embodiment 118. The conjugate of embodiment 117, wherein L-(C) p is the compound-linker construct of any one of embodiments 107-116.

[0719] 119. The conjugate of any one of embodiments 117-118, wherein 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.

[0720] Embodiment 120. The conjugate of embodiment 117, wherein C is the compound of embodiment 36.

[0721] Embodiment 121. The conjugate of embodiment 117, wherein C is the compound of embodiment 66 or embodiment 88.

[0722] Embodiment 122. The conjugate of embodiment 117, wherein C is selected from Sub-Table 1F.

[0723] Embodiment 123. The conjugate of any one of Embodiments 117-122, wherein T binds to a tumor-associated antigen (TAA).

[0724] Embodiment 124. The conjugate of any one of embodiments 117-123, wherein T is an antibody or an antigen-binding antibody fragment.

[0725] Embodiment 125. The conjugate of embodiment 124, wherein the antibody is a bispecific or multispecific antibody.

[0726] Embodiment 126. The conjugate of any one of Embodiments 124-125, wherein the antibody or antigen-binding antibody fragment binds to a TAA.

[0727] Embodiment 127. The conjugate of any one of embodiments 117-126, wherein the conjugate has an EC for inducing cytokine production in a human or murine immune cell population of <1 nM, <500 pM, or <100 pM as determined in a reporter gene assay. 50 value.

[0728] Embodiment 128. A pharmaceutical composition comprising the conjugate of any one of embodiments 117-126 and a pharmaceutically acceptable carrier or diluent.

[0729] Embodiment 129. A method of agonizing TLR7, the method comprising contacting a cell expressing TLR7 with the conjugate of any one of embodiments 117-126, thereby agonizing TLR7.

[0730] Embodiment 130. A method of inducing a cell to release a cytokine, the method comprising contacting the cell with the conjugate of any one of embodiments 117-126, thereby inducing the cell to release the cytokine.

[0731] Embodiment 131. A method of stimulating an immune response in a subject in need thereof, the method comprising administering to the subject an effective amount of the conjugate of any one of embodiments 117-126.

[0732] Embodiment 132. The method of embodiment 132, wherein the conjugate agonizes TLR7 in the subject, thereby stimulating the immune response in the subject.

[0733] Embodiment 133. A method of inhibiting the proliferation of cancer cells, the method comprising contacting a cell population comprising the cancer cells with an effective amount of a conjugate of any one of embodiments 117-126.

[0734] Embodiment 134. A method of killing cancer cells, the method comprising contacting a cell population comprising the cancer cells with an effective amount of a conjugate of any one of embodiments 117-126.

[0735] Embodiment 135. The method of any one of Embodiments 133-134, wherein the cell population comprises immune cells.

[0736] Embodiment 136. A method of treating cancer in a subject in need thereof, comprising administering to the subject an effective amount of the conjugate of any one of embodiments 117-126.

[0737] Embodiment 137. The method of embodiment 136, wherein the conjugate agonizes TLR7 in the subject, thereby treating the cancer in the subject.

[0738] Embodiment 138. The conjugate of any one of embodiments 117-126, for use in therapy.

[0739] Embodiment 139. The conjugate of any one of embodiments 117-126, for use in cancer treatment.

[0740] Embodiment 140. Use of the conjugate of any one of Embodiments 117-126 in the manufacture of a medicament for the treatment of cancer.

[0741] Example

[0742] The following examples are offered for illustrative purposes and are not intended to limit the scope of the present invention in any way.

[0743] The following examples provide illustrative methods for preparing and using compounds of the present disclosure (e.g., compounds of any one of formulas (I)-(IV)). It should be understood that those skilled in the art can synthesize these compounds by similar methods or by combining other methods known in the art. Those skilled in the art can use the methods described herein or similar methods with appropriate starting components and modify synthesis parameters as needed to achieve the preparation of other compounds of formula (I)-(IV) not specifically described herein. Typically, starting components and materials can be obtained from commercial sources 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 synthesized or prepared as described herein according to original data 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).

[0744] 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-aminobenzyloxycarbonyl; PBMC = peripheral blood mononuclear cells; 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-performance liquid chromatography.

[0745] General Chemistry Procedures

[0746] General Procedure 1: Conversion of Chlorides to Amines

[0747] To the chloride compound (50-100 mg / mL) in DMF is added a primary or secondary amine (2-5 equiv) followed by DIPEA (3-5 equiv) and the solution is heated with stirring to 50-75° C. After completion (usually 1-18 h), the reaction mixture is adjusted to pH 2 with 6 M HCl and purified by reverse phase HPLC to give the desired product after lyophilization.

[0748] General Procedure 2: Removal of Boc Protecting Group

[0749] To a stirred solution of the Boc-protected amine compound in dichloromethane (0.1 M) was added TFA (10% by volume). After completion (typically 1 h), the reaction mixture was concentrated in vacuo and co-evaporated with DCM several times to give a crude solid or purified by preparative HPLC to give the desired product after lyophilization.

[0750] General Procedure 3: Formation of Benzyl Chloride

[0751] The alcohol compound is dissolved in 10% SOCl2 / DCM (0.05-0.1M). The solution is stirred at room temperature until completion (usually 12-24 h) and then concentrated in vacuo. The residue is co-evaporated (2×10 mL dichloromethane) to give the crude chloride compound, which is usually used without further purification.

[0752] General Procedure 4: MT-VC-PABC Compound-Linker Construct Synthesis

[0753] To a primary or secondary amine compound (0.05-0.1 M) in dimethylformamide was added the compound of Example 2.5 (MT-VC-PABC-PNP) (1-1.1 equiv) followed by DIPEA (3 equiv). After completion (typically 1-4 h), the reaction mixture was acidified with aqueous HCl (1 M) and then purified by reverse phase HPLC to afford the desired drug linker after lyophilization.

[0754] General Procedure 5: Compound Purification

[0755] Flash chromatography: The crude reaction product was Snap Ultra columns (10 g, 25 g, 50 g, or 100 g) (Biotage, Charlotte, NC) were used and Isolera TM Purification was performed on an automated rapid system (Biotage, Charlotte, NC) using a linear gradient of ethyl acetate / hexane or methanol / dichloromethane. Alternatively, the purified product was purified using Reverse phase flash purification was performed using a Snap Ultra C18 column (12 g, 30 g, 60 g, or 120 g) eluting with a linear gradient of CH3CN+0.1% TFA / H2O+0.1% TFA. The purified compound was isolated by removing the organic solvent via rotary evaporation or lyophilizing the acetonitrile / water mixture.

[0756] Preparative HPLC: The results were analyzed using an Agilent 1260 Infinity II Preparative LC-MSD system (Agilent Technologies, Inc., Santa Clara, CA). 5-μm EVO C18 Reverse phase HPLC of the crude compound was performed using a 250 x 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 an acetonitrile / water mixture.

[0757] General Procedure 6: Compound Analysis

[0758] LC-MS: The reaction was monitored for completion using an Agilent 1290 HPLC / 6120 single quadrupole LC-MS system (Agilent Technologies, Inc., Santa Clara, CA). 2.6-μm C18 The purified compound was analyzed by eluting with a linear gradient of CH3CN+0.1% TFA / H2O+0.1% TFA from 10% to 100% using a 30 x 3 mm column (Phenomenex).

[0759] NMR: 1 H NMR spectra were collected on a Bruker AVANCE III 300 spectrometer (300 MHz) (Bruker Corporation, Billerica, MA). Chemical shifts are expressed in parts per million (ppm).

[0760] General Procedure 7: MT-VK-PABC Compound-Linker Construct Synthesis

[0761] To a primary or secondary amine compound (0.05-0.1 M) in dimethylformamide was added the compound of Example 2.38 (MT-VK(Boc)-PABC-PNP) (1.0-1.1 equiv) followed by DIPEA (3-5 equiv). As indicated, HOBt (1.0 equiv) was used in some reactions. After completion (typically 1-4 h), the reaction mixture was acidified with aqueous HCl (1 M) and then purified by reverse phase HPLC to afford the Boc-protected drug linker after lyophilization. The Boc intermediate was then deprotected according to General Procedure 2. After completion (typically 1 h), the reaction mixture was concentrated in vacuo and co-evaporated with DCM several times to afford a crude solid or purified by preparative HPLC to afford the desired product after lyophilization.

[0762] General Procedure 8: Conversion of mesylate to amine

[0763] To the chloride compound (50-100 mg / mL) in DMF are added a primary or secondary amine (2-5 eq) and NaI (0.1-1.0 eq), followed by DIPEA (3-5 eq), and the solution is heated with stirring to 80° C. After completion (usually 1-18 h), the reaction mixture is adjusted to pH 2 with 6 M HCl and purified by reverse phase HPLC to give the desired product after lyophilization.

[0764] General Procedure 9: Preparation of ISACs using maleimide-containing compound-linker constructs

[0765] 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 thoroughly and incubated at 37°C for 120 min and then cooled on ice. The reduced antibody solution was then stirred for 2 h at 4 °C for 1 h. TMSpin desalting column (40KDa MWCO; ThermoScientific TM ) by further buffer exchange of the antibody solution into 10 mM sodium acetate buffer (pH 4.5). Maleimide functionalized compound-linker construct (10 mM in DMSO) (12-20 equivalents) was added to the reduced protein solution stored on ice. In some embodiments, propylene glycol (10-30% v / v%) was added to the reduced protein solution before the addition of the compound-linker construct. The conjugation reaction was immediately thoroughly mixed with a pipette and the conjugation was allowed to proceed for 120 to 180 min at room temperature. Once completed, the reaction mixture was stirred in a Zebacon® pre-equilibrated with 10 mM sodium acetate (pH 4.5). TM The reaction mixture was purified by passing it through a spin desalting column (40 KDa MWCO; Pierce). The purified conjugate was stored at 4°C and analyzed for total protein content (bicinchoninic acid assay, Pierce micro-BCA protocol, catalog number 23225), characterized by HPLC-HIC, SEC and / or RP-UPLC-MS. The average DAR and drug distribution were derived by interpretation of the HIC and / or LC-MS data. The average DAR of the synthesized conjugates was estimated to be in the range of about 1.5 to 5, or more specifically in the range of 1.9 to 4.3, as further shown herein. nexgen-PTS TM The test system was used with a sensitivity of 0.005 EU / mL. Endotoxin levels were assessed using the LAL test kit (Charles River catalog number PTS20005F). Residual free compound and compound-linker construct levels were assessed by RP-UPLC-MS using a threshold set at 1% ((free compound + compound-linker construct) / (conjugated compound-linker construct)).

[0766] Example 1: Preparation of compounds of formula (I)-(IV)

[0767] Example 1.1: 2,6-Dichloro-9-(tetrahydro-2H-pyran-2-yl)-9H-purine

[0768]

[0769] To a solution of 2,6-dichloro-9H-purine (11 g, 56 mmol, 1.0 equiv) in 100 mL of EtOAc was added dihydropyran (7.0 g, 83 mmol, 1.5 equiv) and p-TsOH (96 mg, 0.56 mmol, 0.010 equiv). The mixture was heated to 50 ° C. for 3 h and then concentrated in vacuo to give the crude title product 1 as a yellow solid (quantitative yield). LC-MS: C 10 H 10 Calculated m / z for Cl2N4O = 272.0, found [M+H] + =273.0.

[0770] Example 1.2: 2-Chloro-9-(tetrahydro-2H-pyran-2-yl)-9H-purin-6-amine

[0771]

[0772] Compound 1 (15 g, 55 mmol) was added to a solution of NH4OH in iPrOH (100 mL, 2.0 M). The mixture was heated to 55 ° C and maintained for 18 h, then poured into H2O (30 mL) and cooled to room temperature. The resulting suspension was filtered and the filter cake was washed with iPrOH (100 mL) to give the title product 2 (13 g, 51 mmol, 93%) as a white solid. LC-MS: C 10 H 10 Calculated m / z for ClN5O = 253.1, found [M+H] + =254.1.

[0773] Example 1.3: 9-Benzyl-2-chloro-9H-purin-6-amine

[0774]

[0775] To a solution of 2-chloroadenine (7.0 g, 41 mmol, 1.0 eq) in DMSO (80 mL) was added KCO (5.7 g, 41 mmol, 1.0 eq) followed by benzyl bromide (7.4 g, 43 mmol, 1.1 eq) and the resulting mixture was stirred at room temperature for 18 h. The reaction was then concentrated in vacuo, the residue dissolved in H2O (100 mL) and EtOAc (50 mL) and the resulting mixture was vigorously stirred for 3 h, after which the EtOAc was removed in vacuo and the suspension was filtered. The filter cake was then washed with H2O (2 x 50 mL) and air-dried for 18 h to afford the title product 3 (7.2 g, 28 mmol, 67%) as a solid. LC-MS: C 12 H 10Calculated m / z for ClN5 = 259.1, found [M+H] + =260.2.

[0776] Example 1.4: 1-((tert-Butyldiphenylsilyl)oxy)pentan-2-ol

[0777]

[0778] To a solution of pentane-1,2-diol (2.9 g, 28 mmol, 1.3 eq) and imidazole (1.5 g, 21 mmol, 1.0 eq) in DCM (10 mL) at 0°C was added TBDPS-Cl, and the mixture was stirred for 1 h. The reaction was then allowed to warm to room temperature and stirred for an additional 36 h, after which it was concentrated in vacuo and the residue was dissolved in EtOAc (30 mL). The resulting organic solution was then washed with brine (2 x 10 mL), dried over MgSO4, and concentrated in vacuo to afford the title product 4 (7.0 g, 20.4 mmol, 95%) as a clear, colorless oil. 1 H NMR (300MHz, CDCl3) δ7.88-7.70(m,5H),7.57-7.35(m,5H),3.89-3.53(m,3H),3.03(s,1H),1.61-1.31(m,4H),1.17(s,9H),1.05-0.92(m,3H).

[0779] Example 1.5: 2-Ethoxy-9-(tetrahydro-2H-pyran-2-yl)-9H-purin-6-amine

[0780]

[0781] 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, after which 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), then dried over MgSO4 and concentrated in vacuo to give the title compound 5 (20.0 g, 76.0 mmol, 96.4%) as an orange solid. LC-MS: C 12 H 17 Calculated m / z for N5O2 = 263.1, found [M+H] + =264.2.

[0782] Example 1.6: 8-Bromo-2-ethoxy-9-(tetrahydro-2H-pyran-2-yl)-9H-purin-6-amine

[0783]

[0784] NBS (20.3 g, 91.2 mmol, 1.20 equiv) was added portionwise to a suspension of compound 5 (20.0 g, 76.0 mmol, 1.00 equiv) in DCM (200 mL) over 3 min. The resulting mixture was stirred at room temperature for 18 h, after which the reaction was quenched by the addition of 1 M NaHSO (50 mL) and the reaction was rapidly stirred for 30 min. The resulting mixture was diluted with DCM (100 mL) and the separated organics were washed with 1 M NaHSO (2×50 mL). The pooled organics were dried over MgSO, filtered, and then concentrated in vacuo to afford the title compound 6 (23.5 g, 68.7 mmol, 90.4%) as an orange solid. LC-MS: C 12 H 16 Calculated m / z for BrN5O2 = 341.1, found [M+H] + =342.2.

[0785] Example 1.7: 2-Ethoxy-8-methoxy-9-(tetrahydro-2H-pyran-2-yl)-9H-purin-6-amine

[0786]

[0787] To a solution of compound 6 (23 g, 67 mmol, 1.0 equiv) in MeOH (150 mL) was added NaOMe (9.1 g, 170 mmol, 2.5 equiv). The resulting mixture was stirred at room temperature for 72 h. NaOMe (3.6 g, 66 mmol, 1.0 equiv) was further added and the mixture was heated to reflux and maintained for 72 h. The solvent was removed in vacuo and the crude material was redissolved in EtOAc (250 mL) and subsequently washed with 1 M NaH2PO4 (150 mL). The aqueous layer was extracted with EtOAc (3×150 mL), the pooled organic layers were dried over MgSO4, and the solution was concentrated in vacuo to give the title compound 7 (17 g, 58 mmol, 86%) as a red solid. LC-MS: C 13 H 19 Calculated m / z for N5O3 = 293.2, found [M+H] + =294.2.

[0788] Example 1.8: 2-Ethoxy-8-methoxy-9-(tetrahydro-2H-pyran-2-yl)-9H-purin-6-amine

[0789]

[0790] Compound 7 (13 g, 44 mmol, 1.0 equiv) was dissolved in 15% TFA / MeOH (40 mL) and the resulting solution was stirred at room temperature for 72 h. The solvent was removed in vacuo to give the title compound 8 (14 g, 40 mmol, 91%) as a yellow solid. LC-MS: C8H 11 Calculated m / z for N5O2 = 209.1, found [M+H] + =210.1.

[0791] Example 1.9: Methyl 4-((6-amino-2-ethoxy-8-methoxy-9H-purin-9-yl)methyl)-3-methoxybenzoate

[0792]

[0793] 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 CsCO (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 accomplished using a 60 g C18 column with a gradient elution from 10% to 60% CHCN + 0.1% TFA / H0 + 0.1% TFA as described in General Procedure 5. The title compound 9 (0.65 g, 1.7 mmol, 50%) was obtained as a white solid. LC-MS: C 18 H 21 Calculated m / z for N5O5 = 387.2, found [M+H] + =388.2.

[0794] Example 1.10: (4-((6-amino-2-ethoxy-8-methoxy-9H-purin-9-yl)methyl)-3-methoxyphenyl)-methanol

[0795]

[0796] Lithium aluminum hydride (24 mg, 0.65 mmol, 1.0 eq) was added in small portions over 5 min to a stirred solution of compound 9 (250 mg, 0.65 mmol, 1.0 eq) in anhydrous THF (5 mL) cooled to 0°C. The resulting suspension was allowed to warm to room temperature and stirred for 15 min, after which it was quenched with H2O (1 mL) and diluted with MeOH (100 mL) and filtered through a plug of celite. The filtrate was concentrated in vacuo and redissolved in 50% CH3CN / H2O (25 mL) to give the title compound 10 (230 mg, 0.63 mmol, 97%) after lyophilization. LC-MS: C 17 H 21 Calculated m / z for N5O4 = 359.2, found [M+H] + =360.2.

[0797] Example 1.11: 6-amino-9-(4-(chloromethyl)-2-methoxybenzyl)-2-ethoxy-9H-purin-8-ol

[0798]

[0799] The title compound 11 was prepared according to General Procedure 3 using compound 10 (210 mg, 0.58 mmol, 1.0 equiv) and 10% SOCl2 / DCM (10 mL). The title compound was obtained as a yellow solid (assuming quantitative yield). LC-MS: C 16 H 18 Calculated m / z for ClN5O3 = 363.1, found [M+H] + =364.2.

[0800] Example 1.12: 6-Amino-2-ethoxy-9-(2-methoxy-4-(piperazin-1-ylmethyl)benzyl)-9H-purin-8-ol (Compound 100)

[0801]

[0802] The title compound was prepared from compound 11 (0.040 g, 0.11 mmol, 1.0 equiv) and tert-butyl piperazine-1-carboxylate (61 mg, 0.33 mmol, 3.0 equiv) according to General Procedure 1. Preparative HPLC purification was accomplished as described in General Procedure 5 with a gradient elution from 10 to 45% CH3CN + 0.1% TFA / H2O + 0.1% TFA. Deprotection was accomplished according to General Procedure 2 to afford the title compound 100 (55 mg, 0.073 mmol, 66%) as a white solid. LC-MS: C 20 H 27Calculated m / z for N7O3 = 413.2, found [M+H] + =414.3. 1 H NMR (300MHz, MeOD) δ7.14(s,1H),7.10(d,J=7.7Hz,1H),6.98(d,J=7.6,1H),5.04(s,2H),4.38(q,J =7.0Hz,2H),4.17(s,2H),3.89(s,3H),3.50-3.42(m,4H),3.33-3.26(m,4H),1.34(t,J=7.1Hz,3H).

[0803] Example 1.13: 6-Amino-2-ethoxy-9-(4-(((2-hydroxy-2-methylpropyl)amino)methyl)-2-methoxybenzyl)-9H-purin-8-ol (Compound 101)

[0804]

[0805] The title compound was prepared from compound 11 (0.010 g, 0.028 mmol, 1.0 equiv) and 1-amino-2-methylpropan-2-ol (7.0 mg, 0.082 mmol, 3.0 equiv) according to General Procedure 1. Preparative HPLC purification was accomplished as described in General Procedure 5 with a gradient elution from 10% to 45% CH3CN + 0.1% TFA / H2O + 0.1% TFA to afford the title compound 101 (11.7 mg, 0.0181 mmol, 66.0%) as a white solid. LC-MS: C 20 H 28 Calculated m / z for N6O4 = 416.2, found [M+H] + =417.3. 1 H NMR(300MHz,CD3CN)δ7.14(s,1H),7.07(d,J=7.8Hz,1H),6.98(d,J=7.71H),4.97(s,2H),4.3 3(q,J=7.1Hz,2H),4.18(s,2H),3.86(s,3H),2.89(s,2H),1.29(t,J=7.1Hz,3H),1.20(s,6H).

[0806] Example 1.14: 6-Amino-9-(4-((diethylamino)methyl)-2-methoxybenzyl)-2-ethoxy-9H-purin-8-ol (Compound 102)

[0807]

[0808] The title compound was prepared from compound 11 (0.010 g, 0.028 mmol, 1.0 equiv) and diethylamine (7.0 mg, 0.082 mmol, 3.0 equiv) according to General Procedure 1. Preparative HPLC purification was accomplished as described in General Procedure 5 with a gradient elution from 10% to 45% CH3CN + 0.1% TFA / H2O + 0.1% TFA to afford the title compound 102 (10 mg, 0.026 mmol, 95%) as a white solid. LC-MS: C 20 H 28 Calculated m / z for N6O3 = 400.2, 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).

[0809] Example 1.15: 6-Amino-2-ethoxy-9-(4-(((5-hydroxypentyl)amino)methyl)-2-methoxybenzyl)-9H-purin-8-ol (Compound 103)

[0810]

[0811] 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 accomplished as described in General Procedure 5 with a gradient elution from 10% to 45% CH3CN + 0.1% TFA / H2O + 0.1% TFA to afford the title compound 103 (10 mg, 0.015 mmol, 95%) as a white solid. LC-MS: C 21 H 30 Calculated m / z for N6O4 = 430.2, found [M+H] + =431.2. 1HNMR(300MHz,CD3CN)δ7.11-7.01(m,2H),6.94(dd,J=7.7,1.6Hz,1H),4.97(s,2H),4.32(q,J=7.0Hz,2H),4.09(s,2H),3.86(s,3H ), 3.49 (t, J = 6.2Hz, 2H), 3.01-2.90 (m, 2H), 1.66 (p, J = 7.7Hz, 2H), 1.49 (p, J = 6.6Hz, 2H), 1.43-1.30 (m, 2H), 1.29 (t, J = 7.1Hz, 3H).

[0812] 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)

[0813]

[0814] The title compound was prepared from compound 11 (0.010 g, 0.028 mmol, 1.0 equiv) and 1,4-piperazine dipropylamine (17 mg, 0.082 mmol, 3.0 equiv) according to General Procedure 1. Preparative HPLC purification was accomplished as described in General Procedure 5 with a gradient elution from 10% to 45% CH3CN + 0.1% TFA / H2O + 0.1% TFA to afford the title compound 104 (14.8 mg, 0.0150 mmol, 54.7%) as a white solid. LC-MS: C 26 H 41 The calculated m / z of N9O3 is 527.3, and the measured [M+H] + =528.3. 1 H NMR(300MHz,CD3CN)δ7.14-7.03(m,2H),6.96(dd,J=7.9,1.5Hz,1H),4.97(s,2H),4.35(q,J=7.1Hz, 2H), 4.13 (s, 2H), 3.85 (s, 3H), 3.13-2.99 (m, 8H), 2.06 (qd, J = 7.9, 2.8Hz, 4H), 1.30 (t, J = 7.0Hz, 3H).

[0815] Example 1.17: 6-Amino-9-(4-(((4-(aminomethyl)benzyl)amino)methyl)-2-methoxybenzyl)-2-ethoxy-9H-purin-8-ol (Compound 105)

[0816]

[0817] The title compound was prepared from compound 11 (0.010 g, 0.028 mmol, 1.0 equiv) and tert-butyl (4-(aminomethyl)benzyl)carbamate (0.020 g, 0.084 mmol, 3.0 equiv) according to General Procedure 1. Preparative HPLC purification was accomplished as described in General Procedure 5 with a gradient elution from 10 to 45% CH3CN + 0.1% TFA / H2O + 0.1% TFA. Half of the resulting Boc intermediate was retained and deprotection of the remaining material was accomplished according to General Procedure 2 to afford the title compound 105 (4.8 mg, 0.0059 mmol, 43%) as a white solid. LC-MS: C 24 H 29 Calculated m / z for N7O3 = 463.2, found [M+H] + =464.3. 1 H NMR(300MHz,CD3CN)δ7.46(s,4H),7.12-7.01(m,2H),6.99-6.91(m,1H),4.96(s,2H),4.33( q,J=7.1Hz,2H),4.18(s,2H),4.17(s,2H),4.11(s,2H),3.85(s,3H),1.29(t,J=7.1Hz,3H).

[0818] Example 1.18: 6-Amino-9-(4-(((4-aminobutyl)amino)methyl)-2-methoxybenzyl)-2-ethoxy-9H-purin-8-ol (Compound 106)

[0819]

[0820] The title compound was prepared from compound 11 (0.010 g, 0.028 mmol, 1.0 equiv) and tert-butyl (4-aminobutyl)carbamate (24 mg, 0.082 mmol, 3.0 equiv) according to General Procedure 1. Preparative HPLC purification was accomplished as described in General Procedure 5 with a gradient elution from 10 to 45% CH3CN + 0.1% TFA / H2O + 0.1% TFA. Half of the resulting Boc intermediate was retained and deprotection of the remaining material was accomplished according to General Procedure 2 to afford the title compound 106 (5.6 mg, 0.0050 mmol, 36%) as a white solid. LC-MS: C 20 H 29 Calculated m / z for N7O3 = 415.2, found [M+H] + =416.3. 1H NMR(300MHz,CD3CN)δ7.13-6.99(m,2H),6.95(dd,J=7.8,1.6Hz,1H),4.96(s,2H),4.29(q,J=7.1Hz,2H) ,4.11(s,2H),3.86(s,3H),3.04-2.86(m,4H),1.75-1.60(dq,J=12.5,6.5Hz,4H),1.29(t,J=7.0Hz,3H).

[0821] Example 1.19: 6-Amino-9-(4-((Benzylamino)methyl)-2-methoxybenzyl)-2-ethoxy-9H-purin-8-ol (Compound 107)

[0822]

[0823] The title compound was prepared from compound 11 (0.010 g, 0.028 mmol, 1.0 equiv) and benzylamine (9.0 mg, 0.082 mmol, 3.0 equiv) according to General Procedure 1. Preparative HPLC purification was accomplished as described in General Procedure 5 with a gradient elution from 10% to 45% CH3CN + 0.1% TFA / H2O + 0.1% TFA to afford the title compound 107 (14 mg, 0.021 mmol, 76%) as a white solid. LC-MS: C 23 H 26 Calculated m / z for N6O3 = 434.2, found [M+H] + =435.3. 1 H NMR(300MHz,CD3CN)δ7.45(s,5H),7.11-6.99(m,2H),6.95(dd,J=7.7,1.6Hz,1H),4.96(s ,2H),4.30(q,J=7.1Hz,2H),4.17(s,2H),4.15(s,2H),3.86(s,3H),1.29(t,J=7.0Hz,3H).

[0824] Example 1.20: 6-Amino-9-(4-((cyclobutylamino)methyl)-2-methoxybenzyl)-2-ethoxy-9H-purin-8-ol (Compound 193)

[0825]

[0826] The title compound was prepared from compound 11 (0.010 g, 0.028 mmol, 1.0 equiv) and pyrcosamine (15 mg, 0.082 mmol, 3.0 equiv) according to General Procedure 1. Preparative HPLC purification was accomplished as described in General Procedure 5 with a gradient elution from 10% to 45% CH3CN + 0.1% TFA / H2O + 0.1% TFA to afford the title compound 193 (15 mg, 0.020 mmol, 72%) as a white solid. LC-MS: C 22 H 25 Calculated m / z for N7O3 = 435.2, found [M+H] + =436.3. 1 H NMR(300MHz,CD3CN)δ8.80(s,1H),8.74-8.68(m,1H),8.36(dd,J=8.1,2.1Hz,1H),7.82(dd,J=8.1,5.4Hz,1H),7.11(d,J=1.6Hz, 1H),7.05(d,J=7.8Hz,1H),6.97(d,J=7.8,1H),4.96(s,2H),4.37-4.29(m,4H),4.24(s,2H),3.84(s,3H),1.29(t,J=7.0Hz,3H).

[0827] Example 1.21: 6-Amino-9-(4-((cyclobutylamino)methyl)-2-methoxybenzyl)-2-ethoxy-9H-purin-8-ol (Compound 108)

[0828]

[0829] The title compound was prepared from compound 11 (0.010 g, 0.028 mmol, 1.0 equiv) and cyclobutylamine (0.010 g, 0.082 mmol, 3.0 equiv) according to General Procedure 1. Preparative HPLC purification was accomplished as described in General Procedure 5 with a gradient elution from 10% to 45% CH3CN + 0.1% TFA / H2O + 0.1% TFA to afford the title compound 108 (9.0 mg, 0.014 mmol, 52%) as a white solid. LC-MS: C 20 H 26 Calculated m / z for N6O3 = 398.2, found [M+H] + =399.3. 1H NMR (300MHz, CD3CN) δ7.09-7.00(m,2H),6.92(dd,J=7.8,1.6Hz,1H),4.96(s,2H),4.30(q,J=7.1Hz,2H),3. 99(s,2H),3.86(s,3H),3.69(p,J=8.2Hz,1H),2.28-2.09(m,4H),1.95-1.73(m,2H),1.29(t,J=7.0Hz,3H).

[0830] Example 1.22: 6-Amino-9-(4-(((3,3-difluorocyclobutyl)amino)methyl)-2-methoxybenzyl)-2-ethoxy-9H-purin-8-ol (Compound 109)

[0831]

[0832] The title compound was prepared from compound 11 (0.010 g, 0.028 mmol, 1.0 equiv) and 3,3-difluorocyclobutanamine (15 mg, 0.12 mmol, 3.0 equiv) according to General Procedure 1. Preparative HPLC purification was accomplished as described in General Procedure 5 with a gradient elution from 10% to 45% CH3CN + 0.1% TFA / H2O + 0.1% TFA to afford the title compound 109 (12 mg, 0.018 mmol, 66%) as a white solid. LC-MS: C 20 H 24 The calculated m / z of F2N6O3 is 434.2, and the measured [M+H] + =435.3. 1 H NMR(300MHz,MeOD)δ7.14(s,1H),7.08(d,J=7.6Hz,1H),7.00(d,J=8.2Hz,1H),5.05(s,2H),4.31(q, J=5.2Hz,2H),4.19(s,2H),3.93(s,3H),3.87-3.75(m,1H),3.18-2.76(m,4H),1.33(t,J=7.1Hz,3H).

[0833] Example 1.23: 6-Amino-9-(4-((4-aminopiperidin-1-yl)methyl)-2-methoxybenzyl)-2-ethoxy-9H-purin-8-ol (Compound 110)

[0834]

[0835] The title compound was prepared from compound 11 (15.0 mg, 0.0412 mmol, 1.00 equiv) and 4-Boc-aminopiperidine (25 mg, 0.12 mmol, 3.0 equiv) according to General Procedure 1. Preparative HPLC purification was accomplished as described in General Procedure 5 with a gradient elution from 10 to 45% CH3CN + 0.1% TFA / H2O + 0.1% TFA. Half of the resulting Boc intermediate was retained and deprotection of the remaining material was accomplished according to General Procedure 2 to afford the title compound 110 (8.2 mg, 0.011 mmol, 52%) as a white solid. LC-MS: C 21 H 29 Calculated m / z for N7O3 = 427.2, found [M+H] + =428.3. 1 H NMR(300MHz,CD3CN)δ7.14-7.11(m,2H),6.97(dd,J=7.7,1.6Hz,1H),4.99(s,2H),4.37(q,J=7.1Hz,2H), 4.21(s,2H),3.86(s,3H),3.50-3.41(m,4H),3.07-2.96(m,2H),2.25-2.18(m,2H),1.30(t,J=7.1Hz,3H).

[0836] Example 1.24: 6-Amino-9-(4-(((2-aminoethyl)amino)methyl)-2-methoxybenzyl)-2-ethoxy-9H-purin-8-ol (Compound 111)

[0837]

[0838] The title compound was prepared from compound 11 (15 mg, 0.041 mmol, 1.0 equiv) and tert-butyl (2-aminoethyl)carbamate (20 mg, 0.12 mmol, 3.0 equiv) according to General Procedure 1. Preparative HPLC purification was accomplished as described in General Procedure 5 with a gradient elution from 10 to 45% CH3CN + 0.1% TFA / H2O + 0.1% TFA. Half of the resulting Boc intermediate was retained and deprotection of the remaining material was accomplished according to General Procedure 2 to afford the title compound 111 (11.0 mg, 0.0151 mmol, 73.1%) as a white solid. LC-MS: C 18 H 25 The calculated m / z of N7O3 is 387.2, and the measured [M+H] + =388.3. 1H NMR(300MHz,CD3CN)δ7.11(m,2H),6.97(dd,J=7.8,1.6Hz,1H),4.98(s,2H),4.39(q ,J=7.0Hz,2H),4.18(s,2H),3.85(s,3H),3.43-3.29(m,4H),1.31(t,J=7.1Hz,3H).

[0839] Example 1.25: 6-amino-9-(4-(((2-aminoethyl)amino)methyl)-2-methoxybenzyl)-2-ethoxy-9H-purin-8-ol (Compound 112)

[0840]

[0841] The title compound was prepared from compound 11 (15 mg, 0.041 mmol, 1.0 equiv) and tert-butyl (2-(piperazin-1-yl)ethyl)carbamate (28 mg, 0.12 mmol, 3.0 equiv) according to General Procedure 1. Preparative HPLC purification was accomplished as described in General Procedure 5 with a gradient elution from 10 to 45% CH3CN + 0.1% TFA / H2O + 0.1% TFA. Half of the resulting Boc intermediate was retained and deprotection of the remaining material was accomplished according to General Procedure 2 to afford the title compound 112 (17 mg, 0.019 mmol, 91%) as a white solid. LC-MS: C 22 H 32 Calculated m / z for N8O3 = 456.2, found [M+H] + =457.3. 1 H NMR (300MHz, CD3CN) δ7.20-7.07(m,1H),7.12(s,1H),6.99(d,J=7.5Hz,1H),4.99(s,2H),4.40(q,J=7.1,2H), 4.24(s,2H),3.84(s,3H),3.42-3.35(m,8H),3.07(q,J=7.0Hz,2H),2.89-2.73(m,2H),1.31(t,J=7.0Hz,3H).

[0842] 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)

[0843]

[0844] To a solution of Example 2.26 (35 mg, 0.046 mmol, 1.0 equiv) in DMF (500 μL) and CH3CN (500 μL) was added HO (2 mL) followed by L-cysteine ​​(17 mg, 0.14 mmol, 3.0 equiv) and 0.5 M Na2HPO4 (500 μL). The resulting solution was stirred at room temperature for 1 h before 10 M NaOH (600 μL) was added and the reaction mixture was stirred at room temperature for an additional 2 h. Reverse phase flash purification was accomplished as described in General Procedure 5 using a 12 g C18 column eluting with a gradient of 10% to 50% CH3CN+0.1%TFA / HO+0.1%TFA to afford the title compound 113 (35 mg, 0.031 mmol, 68%) as a white solid. LC-MS: C 29 H 41 Calculated m / z for N9O8S = 675.3, found [M+H] + =676.4. 1 H NMR(300MHz,MeOD)δ7.19-7.06(m,2H),6.99(d,J=7.7Hz,1H),5.05(s,2H),4.51-4.21(m,3H),4.19(d,J=4.0Hz,2H),3.9 0(s,3H),3.88-3.71(m,1H),3.64-2.99(m,14H),2.87(dd,J=16.4,7.1Hz,1H),2.77-2.50(m,1H),1.35(t,J=7.1Hz,3H).

[0845] Example 1.27: N 6 -(4-((6-amino-2-ethoxy-8-hydroxy-9H-purin-9-yl)methyl)-3-methoxybenzyl)-L-lysine (Compound 114)

[0846]

[0847] To N αTo a solution of -Boc-L-lysine (32 mg, 0.14 mmol, 5.0 equiv) in aqueous NaHCO3 (0.275 mL, 1.14 M, 12.5 equiv) was added a solution of compound 11 (0.010 g, 0.027 mmol, 1.0 equiv) in 100 μL DMA. The resulting solution was heated to 50° C. and maintained for 4 h, then acidified with aqueous HCl (5 M). Preparative HPLC purification was accomplished as described in General Procedure 5 with a gradient elution of 10 to 45% CH3CN+0.1%TFA / H2O+0.1%TFA. Deprotection was accomplished according to General Procedure 2 to afford the title compound 114 (7.9 mg, 0.010 mmol, 37%) as a white solid. LC-MS: C 22 H 31 Calculated m / z for N7O5 = 473.2, found [M+H] + =474.3. 1 H NMR (300MHz, MeOD) δ7.18-7.04(m,2H),7.00(d,J=7.7Hz,1H),5.05(s,2H),4.34(q,J=7.1Hz,2H),4.20( s,2H),3.98(t,J=6.3Hz,1H),3.93(s,3H),3.13-3.01(m,2H),2.05-1.48(m,6H),1.34(t,J=7.1Hz,4H).

[0848] Example 1.28: 6-((4-((6-Amino-2-ethoxy-8-hydroxy-9H-purin-9-yl)methyl)-3-methoxybenzyl)-amino)hexanoic acid (Compound 115)

[0849]

[0850] 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) was added a solution of compound 11 (0.010 g, 0.014 mmol, 1.0 equiv) in 100 μL DMA. The resulting solution was heated to 50° C. and maintained for 4 h, then acidified with aqueous HCl (5 M). Preparative HPLC purification was accomplished as described in General Procedure 5 with a gradient elution of 10% to 45% CH3CN+0.1% TFA / H2O+0.1% TFA to afford the title compound 115 (6.7 mg, 0.013 mmol, 48%) as a white solid. LC-MS: C 22 H 30 Calculated m / z for N6O5 = 458.2, found [M+H] +=459.2. 1 H NMR(300MHz,MeOD)δ7.14(d,J=7.5Hz,2H),7.01(dd,J=7.6,1.6Hz,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=7.2Hz,2H),1.79-1.61(m,4H),1.52-1.25(m,5H).

[0851] Example 1.29: (3-((4-((6-amino-2-ethoxy-8-hydroxy-9H-purin-9-yl)methyl)-3-methoxybenzyl)-amino)-propionyl)-L-histidine (Compound 116)

[0852]

[0853] To a solution of carnosine (31 mg, 0.14 mmol, 5.0 equiv) in aqueous NaHCO3 (0.275 mL, 1.14 M, 12.5 equiv) was added a solution of compound 11 (0.010 g, 0.014 mmol, 1.0 equiv) in 100 μL DMA. The resulting solution was heated to 50° C. and maintained for 4 h, then acidified with aqueous HCl (5 M). Preparative HPLC purification was accomplished as described in General Procedure 5 with a gradient elution from 10% to 45% CH3CN+0.1% TFA / H2O+0.1% TFA to afford the title compound 116 (8.5 mg, 0.0090 mmol, 35%) as a white solid. LC-MS: C 25 H 31 Calculated m / z for N9O6 = 553.2, found [M+H] + =554.3. 1 H NMR(300MHz,MeOD)δ8.83(d,J=1.4Hz,1H),7.37(s,1H),7.18-6.98(m,3H),5.06(s,2H),4.80(dd,J=8.4,5.3Hz,1H),4 .38(q,J=7.1Hz,2H),4.22(s,2H),3.93(s,3H),3.41-3.05(m,4H),2.72(dd,J=7.6,5.7Hz,2H),1.36(t,J=7.1Hz,3H).

[0854] Example 1.30: (4-((6-amino-2-ethoxy-8-hydroxy-9H-purin-9-yl)methyl)-3-methoxybenzyl)-L-phenylalanine (Compound 117)

[0855]

[0856] To a solution of L-phenylalanine (22 mg, 0.14 mmol, 5.0 equiv) in aqueous NaHCO3 (0.275 mL, 1.14 M, 12.5 equiv) was added a solution of compound 11 (0.010 g, 0.014 mmol, 1.0 equiv) in 100 μL DMA. The resulting solution was heated to 50° C. and maintained for 4 h, then acidified with aqueous HCl (5 M). Preparative HPLC purification was accomplished as described in General Procedure 5 with a gradient elution from 10% to 45% CH3CN+0.1% TFA / H2O+0.1% TFA to afford the title compound 117 (6.2 mg, 0.0086 mmol, 32%) as a white solid. LC-MS: C 25 H 28 Calculated m / z for N6O5 = 492.2, found [M+H] + =493.3. 1 H NMR(300MHz,MeOD)δ7.39-7.24(m,5H),7.12-7.07(m,2H),6.98(d,J=7.7Hz,1H),5.06(s,2H),4 .37(q,J=7.1Hz,2H),4.28-4.19(m,3H),3.91(s,3H),3.29-3.21(m,2H),1.35(t,J=7.2Hz,3H).

[0857] 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)

[0858]

[0859] To a solution of L-phenylalaninol (21 mg, 0.14 mmol, 5.0 equiv) in aqueous NaHCO3 (0.275 mL, 1.14 M, 12.5 equiv) was added a solution of compound 11 (0.010 g, 0.014 mmol, 1.0 equiv) in 100 μL DMA. The resulting solution was heated to 50° C. and maintained for 4 h, then acidified with aqueous HCl (5 M). Preparative HPLC purification was accomplished as described in General Procedure 5 with a gradient elution of 10% to 45% CH3CN+0.1%TFA / H2O+0.1%TFA to afford the title compound 118 (11 mg, 0.015 mmol, 55%) as a white solid. LC-MS: C 25 H 30Calculated m / z for N6O4 = 478.2, found [M+H] + =479.3. 1 H NMR (300MHz, MeOD) δ7.41-7.11(m,7H),7.04(dd,J=7.8,1.6Hz,1H),5.07(s,2H),4.45-4.22(m,4H),3.93(s,3H),3.78(dd,J=12.1,3.3Hz ,1H),3.58(dd,J=12.2,4.7Hz,1H),3.50-3.35(m,1H),3.11(dd,J=13.5,4.7Hz,1H),2.99(dd,J=13.5,10.3Hz,1H),1.36(t,J=7.1Hz,3H).

[0860] Example 1.32: (4-((6-amino-2-ethoxy-8-hydroxy-9H-purin-9-yl)methyl)-3-methoxybenzyl)-L-serine (Compound 119)

[0861]

[0862] To a solution of L-serine (14 mg, 0.14 mmol, 5.0 equiv) in aqueous NaHCO3 (0.275 mL, 1.14 M, 12.5 equiv) was added a solution of compound 11 (0.010 g, 0.014 mmol, 1.0 equiv) in 100 μL DMA. The resulting solution was heated to 50° C. and maintained for 4 h, then acidified with aqueous HCl (5 M). Preparative HPLC purification was accomplished as described in General Procedure 5 with a gradient elution from 10% to 45% CH3CN+0.1% TFA / H2O+0.1% TFA to afford the title compound 119 (8.7 mg, 0.013 mmol, 48%) as a white solid. LC-MS: C 19 H 24 Calculated m / z for N6O6 = 432.2, found [M+H] + =433.3. 1 H NMR(300MHz,MeOD)δ7.22-7.12(m,2H),7.04(dd,J=7.7,1.6Hz,1H),5.08(s,2H),4.43 (q,J=7.1Hz,2H),4.31(s,2H),4.15-3.98(m,3H),3.93(s,3H),1.38(t,J=7.1Hz,3H).

[0863] 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)

[0864]

[0865] To a solution of iminodiacetic acid (18 mg, 0.14 mmol, 5.0 equiv) in aqueous NaHCO3 (0.275 mL, 1.14 M, 12.5 equiv) was added a solution of compound 11 (0.010 g, 0.014 mmol, 1.0 equiv) in 100 μL DMA. The resulting solution was heated to 50° C. and maintained for 4 h, then acidified with aqueous HCl (5 M). Preparative HPLC purification was accomplished as described in General Procedure 5 with a gradient elution of 10% to 45% CH3CN+0.1%TFA / H2O+0.1%TFA to afford the title compound 120 (7.9 mg, 0.011 mmol, 42%) as a white solid. LC-MS: C 20 H 24 Calculated m / z for N6O7 = 460.2, found [M+H] + =461.3. 1 H NMR (300MHz, MeOD) δ7.22-6.99(m,3H),5.07(s,2H),4.51(s,2H),4.40-4.34(m,2H),4.16-4.08(m,4H),3.94(s,3H),1.35(t,J=7.1,3H).

[0866] Example 1.34: (4-((6-amino-2-ethoxy-8-hydroxy-9H-purin-9-yl)methyl)-3-methoxybenzyl)-L-isoleucine (Compound 121)

[0867]

[0868] To a solution of (2S, 3S)-2-amino-3-hydroxypentanoic acid (37 mg, 0.28 mmol, 10 equiv) in aqueous NaHCO3 (0.362 mL, 1.14 M, 15.0 equiv) was added a solution of compound 11 (0.010 g, 0.014 mmol, 1.0 equiv) in 100 μL DMA. The resulting solution was heated to 50 ° C and maintained for 18 h, then acidified with aqueous HCl (1 M). Preparative HPLC purification was completed as described in General Procedure 5 with a gradient elution of 10% to 45% CH3CN + 0.1% TFA / H2O + 0.1% TFA to give the title compound 121 (7.9 mg, 0.012 mmol, 42%) as a white solid. LC-MS: C 22 H 30 Calculated m / z for N6O5 = 458.2, found [M+H] + =459.2. 1 HNMR(300MHz,MeOD)δ7.19-7.13(m,1H),7.12-6.96(m,2H),5.05(s,2H),4.40-4.17(m,4H),3.96-3.85(m ,1H),2.10-1.93(m,1H),1.72-1.53(m,1H),1.53-1.38(m,1H),1.34(t,J=7.1Hz,3H),1.06-0.92(m,6H).

[0869] 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)

[0870]

[0871] The title compound was prepared from compound 11 (0.010 g, 0.027 mmol, 1.0 equiv) and (2R,3S)-2-aminopentane-1,3-diol (16 mg, 0.14 mmol, 5.0 equiv) according to General Procedure 1. Preparative HPLC purification was accomplished as described in General Procedure 5 with a gradient elution from 10% to 45% CH3CN + 0.1% TFA / H2O + 0.1% TFA to afford the title compound 122 (9.8 mg, 0.015 mmol, 53%) as a white solid. LC-MS: C 22 H 32 Calculated m / z for N6O4 = 444.2, found [M+H] + =445.3. 1H NMR(300MHz,MeOD)δ7.18(s,1H),7.12-6.98(m,2H),5.04(s,2H),4.41-4.22(m,4H),3.94-3.81(m,1H),3.75(dd,J=12 .1,7.1Hz,1H),3.19-3.08(m,1H),1.94-1.80(m,1H),1.54-1.18(m,5H),0.98(d,J=6.9Hz,3H),0.90(t,J=7.4Hz,3H).

[0872] Example 1.36: 6-Amino-9-(4-(aminomethyl)-2-methoxybenzyl)-2-ethoxy-9H-purin-8-ol (Compound 123)

[0873]

[0874] The title compound was prepared from compound 11 (0.010 g, 0.027 mmol, 1.0 equiv) and ammonia / DMF (15 μL, 2 M, 5.0 equiv) according to General Procedure 1. Preparative HPLC purification was accomplished as described in General Procedure 5 with a gradient elution from 10% to 45% CH 3 CN + 0.1% TFA / H 2 O + 0.1% TFA to afford the title compound 123 as a white solid (1.7 mg, 0.0030 mmol, 11%). LC-MS: C 16 H 20 The calculated m / z of N6O3 is 344.2, and the measured [M+H] + =345.2. 1 H NMR (300MHz, MeOD) δ7.13-6.99 (m, 2H), 6.95 (d, J = 7.5Hz, 1H), 5.04 (s, 2H), 4.30 (q, J = 7.1Hz, 2H), 4.10 (s, 2H), 3.93 (s, 3H), 1.33 (t, J = 7.0Hz, 3H).

[0875] Example 1.37: 6-Amino-2-ethoxy-9-(4-((ethylamino)methyl)-2-methoxybenzyl)-9H-purin-8-ol (Compound 124)

[0876]

[0877] The title compound was prepared from compound 11 (0.010 g, 0.027 mmol, 1.0 equiv) and ethylamine (9.3 μL, 0.14 mmol, 5.0 equiv) according to General Procedure 1. Preparative HPLC purification was accomplished as described in General Procedure 5 with a gradient elution from 10% to 45% CH 3 CN + 0.1% TFA / H 2 O + 0.1% TFA to afford the title compound 124 (6.4 mg, 0.011 mmol, 39%) as a white solid. LC-MS: C 16 H 20 Calculated m / z for N6O3 = 372.2, found [M+H] + =373.2. 1 H NMR(300MHz,MeOD)δ7.17-7.04(m,2H),7.00(dd,J=7.7,1.6Hz,1H),5.05(s,2H),4.34( q,J=7.1Hz,2H),4.18(s,2H),3.93(s,3H),3.12(q,J=7.3Hz,2H),1.34(t,J=7.2Hz,6H).

[0878] Example 1.38: 6-Amino-9-(4-((dipropylamino)methyl)-2-methoxybenzyl)-2-ethoxy-9H-purin-8-ol (Compound 125)

[0879]

[0880] The title compound was prepared from compound 11 (0.010 g, 0.027 mmol, 1.0 equiv) and dipropylamine (19 μL, 0.14 mmol, 5.0 equiv) according to General Procedure 1. Preparative HPLC purification was accomplished as described in General Procedure 5 with a gradient elution from 10% to 45% CH 3 CN + 0.1% TFA / H 2 O + 0.1% TFA to afford the title compound 125 (2.5 mg, 0.0038 mmol, 14%) as a white solid. LC-MS: C 22 H 32 Calculated m / z for N6O3 = 428.3, found [M+H] + =429.3. 1 H NMR(300MHz,MeOD)δ7.15(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=8.5Hz,4H),1.92-1.65(m,4H),1.33(t,J=7.1Hz,3H),1.00(t,J=7.3Hz,6H).

[0881] 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)

[0882]

[0883] 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 accomplished as described in General Procedure 5 with a gradient elution from 10 to 45% CH3CN + 0.1% TFA / H2O + 0.1% TFA. Half of the resulting Boc intermediate was retained and deprotection of the remaining material was accomplished according to General Procedure 2 to afford the title compound 126 (3.0 mg, 0.0070 mmol, 26%) as a white solid. LC-MS: C 20 H 27 Calculated m / z for N7O3 = 413.2, found [M+H] + =414.3. 1 H NMR (300MHz, MeOD) δ7.13 (d, J=1.6Hz, 1H), 7.08-6.94 (m, 2H), 6.15-5.92 (m, 2H), 5.04 (s, 2H), 4.29 (q, J= 7.1Hz, 2H), 4.19 (s, 2H), 3.93 (s, 3H), 3.76 (d, J = 5.8Hz, 2H), 3.66 (d, J = 5.4Hz, 2H), 1.32 (t, J = 7.1Hz, 3H).

[0884] Example 1.40: 6-Amino-2-ethoxy-9-(2-methoxy-4-((piperidin-3-ylamino)methyl)benzyl)-9H-purin-8-ol (Compound 127)

[0885]

[0886] The title compound was prepared from compound 11 (0.010 g, 0.028 mmol, 1.0 equiv) and tert-butyl 3-aminopiperidine-1-carboxylate (30 mg, 0.15 mmol, 5.0 equiv) according to General Procedure 1. Preparative HPLC purification was accomplished as described in General Procedure 5 with a gradient elution from 10 to 45% CH3CN + 0.1% TFA / H2O + 0.1% TFA. Half of the resulting Boc intermediate was retained and deprotection of the remaining material was accomplished according to General Procedure 2 to afford the title compound 127 (3.8 mg, 0.0088 mmol, 63%) as a white solid. LC-MS: C 21 H 29 Calculated m / z for N7O3 = 427.2, found [M+H] + =428.2. 1 H NMR(300MHz,MeOD)δ7.18(s,1H),7.10-6.97(m,2H),5.04(s,2H),4.38-4.22(m,4H),3.94(s,3H),3.92-3.70(m,2H), 3.62-3.38(m,2H),3.15-2.91(m,2H),2.45-2.35(m,1H),2.21-2.10(m,1H),1.88-1.55(m,1H),1.33(t,J=7.1Hz,3H).

[0887] Example 1.41: 9-(4-(((1S,4S)-2,5-diazabicyclo[2.2.1]hept-2-yl)methyl)-2-methoxybenzyl)-6-amino-2-ethoxy-9H-purin-8-ol (Compound 128)

[0888]

[0889] The title compound was prepared from compound 11 (0.010 g, 0.028 mmol, 1.0 equiv) and (1S,4S)-2,5-diazabicyclo[2.2.1]heptane-2-carboxylic acid tert-butyl ester (27 mg, 0.15 mmol, 5.0 equiv) according to General Procedure 1. Preparative HPLC purification was accomplished as described in General Procedure 5 with a gradient elution from 10 to 45% CH3CN+0.1%TFA / H2O+0.1%TFA. Half of the resulting Boc intermediate was retained and deprotection of the remaining material was accomplished according to General Procedure 2 to afford the title compound 128 (8.0 mg, 0.010 mmol, 71%) as a white solid. LC-MS: C 21 H 27 Calculated m / z for N7O3 = 425.2, found [M+H] + =426.3. 1H NMR (300MHz, MeOD) δ7.20 (s, 1H), 7.10-6.98 (m, 2H), 5.04 (s, 2H), 4.52 (s, 1H), 4.42-4.27 (m, 4H), 4.24 (d, J = 13.0Hz, 1H), 3. 93(s,3H),3.75(d,J=12.8Hz,1H),3.54-3.35(m,3H),2.58(d,J=12.7Hz,1H),2.16(d,J=12.6Hz,1H),1.34(t,J=7.1Hz,3H).

[0890] Example 1.42: 6-Amino-9-(4-((azetidin-3-ylamino)methyl)-2-methoxybenzyl)-2-ethoxy-9H-purin-8-ol (Compound 129)

[0891]

[0892] The title compound was prepared from compound 11 (0.010 g, 0.028 mmol, 1.0 equiv) and tert-butyl 3-aminoazetidine-1-carboxylate (20 mg, 0.09 mmol, 3 equiv) according to General Procedure 1. Preparative HPLC purification was accomplished as described in General Procedure 5 with a gradient elution from 10 to 45% CH3CN + 0.1% TFA / H2O + 0.1% TFA. Half of the resulting Boc intermediate was retained and deprotection of the remaining material was accomplished according to General Procedure 2 to afford the title compound 129 (7.0 mg, 0.0094 mmol, 67%) as a white solid. LC-MS: C 19 H 25 Calculated m / z for N7O3 = 399.2, found [M+H] + =400.3. 1 H NMR(300MHz,MeOD)δ7.16(s,1H),7.04(d,J=7.7Hz,1H),7.01-6.93(m,1H),5 .04(s,2H),4.37-4.20(m,6H),4.15(s,2H),3.94(s,3H),1.44-1.28(m,4H).

[0893] Example 1.43: 9-(4-(((2-Azaspiro[3.3]heptan-6-yl)amino)methyl)-2-methoxybenzyl)-6-amino-2-ethoxy-9H-purin-8-ol (Compound 130)

[0894]

[0895] The title compound was prepared from compound 11 (0.010 g, 0.028 mmol, 1.0 equiv) and tert-butyl 6-amino-2-azaspiro[3.3]heptane-2-carboxylate (20 mg, 0.09 mmol, 3 equiv) according to General Procedure 1. Preparative HPLC purification was accomplished as described in General Procedure 5 with a gradient elution from 10 to 45% CH3CN+0.1%TFA / H2O+0.1%TFA. Half of the resulting Boc intermediate was retained and deprotection of the remaining material was accomplished according to General Procedure 2 to afford the title compound 130 (7.9 mg, 0.010 mmol, 72%) as a white solid. LC-MS: C 22 H 29 Calculated m / z for N7O3 = 439.2, found [M+H] + =440.3. 1 HNMR(300MHz,MeOD)δ7.25-7.02(m,2H),6.97(dd,J=7.7,1.6Hz,1H),5.05(s,2H),4.34(q,J=7.1Hz,2H),4.17(s,2H) ,4.09(d,J=3.2Hz,4H),3.93(s,3H),3.78-3.65(m,1H),2.81-2.62(m,2H),2.58-2.45(m,2H),1.35(t,J=7.1Hz,3H).

[0896] 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)

[0897]

[0898] The title compound was prepared from compound 11 (0.010 g, 0.028 mmol, 1.0 equiv) and tert-butyl 4-(3-aminopropyl)piperazine-1-carboxylate (20 mg, 0.09 mmol, 3 equiv) according to General Procedure 1. Preparative HPLC purification was accomplished as described in General Procedure 5 with a gradient elution from 10 to 45% CH3CN + 0.1% TFA / H2O + 0.1% TFA. Half of the resulting Boc intermediate was retained and deprotection of the remaining material was accomplished according to General Procedure 2 to afford the title compound 131 (0.010 g, 0.011 mmol, 72%) as a white solid. LC-MS: C 23 H 34 Calculated m / z for N8O3 = 470.3, found [M+H] + =471.4. 1H NMR(300MHz,MeOD)δ7.28-6.97(m,3H),5.06(s,2H),4.45-4.28(m,2H),4.22(s,2H),3.93(s,3H),3.32-3.23( m,4H),3.27-3.10(m,2H),2.97-2.82(m,4H),2.71(t,J=6.8Hz,2H),2.08-1.89(m,2H),1.36(t,J=7.0Hz,3H).

[0899] Example 1.45: 6-amino-9-(4-((4-(aminomethyl)piperidin-1-yl)methyl)-2-methoxybenzyl)-2-ethoxy-9H-purin-8-ol (Compound 132)

[0900]

[0901] The title compound was prepared from compound 11 (0.010 g, 0.028 mmol, 1.0 equiv) and tert-butyl N-(piperidin-4-ylmethyl)carbamate (20 mg, 0.09 mmol, 3 equiv) according to General Procedure 1. Preparative HPLC purification was accomplished as described in General Procedure 5 with a gradient elution from 10 to 45% CH3CN + 0.1% TFA / H2O + 0.1% TFA. Half of the resulting Boc intermediate was retained and deprotection of the remaining material was accomplished according to General Procedure 2 to afford the title compound 132 (7.9 mg, 0.010 mmol, 72%) as a white solid. LC-MS: C 22 H 31 Calculated m / z for N7O3 = 441.3, found [M+H] + =442.4. 1 H NMR(300MHz,MeOD)δ7.20-6.97(m,3H),5.06(s,2H),4.41-4.28(m,4H),3.94(s,3H),3.57(d,J=12.4Hz,2 H), 3.11-2.96 (m, 2H), 2.91 (d, J = 6.7Hz, 2H), 2.25-1.86 (m, 3H), 1.65-1.51 (m, 2H), 1.34 (t, J = 7.1Hz, 3H).

[0902] Example 1.46: (S)-6-Amino-2-ethoxy-9-(2-methoxy-4-((pyrrolidin-3-ylamino)methyl)benzyl)-9H-purin-8-ol (Compound 133)

[0903]

[0904] The title compound was prepared from compound 11 (0.010 g, 0.028 mmol, 1.0 equiv) and tert-butyl (3R)-3-aminopyrrolidine-1-carboxylate (20 mg, 0.09 mmol, 3 equiv) according to General Procedure 1. Preparative HPLC purification was accomplished as described in General Procedure 5 with a gradient elution from 10 to 45% CH3CN + 0.1% TFA / H2O + 0.1% TFA. Half of the resulting Boc intermediate was retained and deprotection of the remaining material was accomplished according to General Procedure 2 to afford the title compound 133 (0.010 g, 0.013 mmol, 93%) as a white solid. LC-MS: C 20 H 27 Calculated m / z for N7O3 = 413.2, found [M+H] + =414.3. 1 H NMR(300MHz,MeOD)δ7.21(s,1H),7.15-6.99(m,2H),5.06(s,2H),4.41-4.27(m,4H),4.14(p,J=7.1Hz,1H),3.94(s,3H) ,3.81-3.68(m,1H),3.67-3.48(m,2H),3.48-3.33(m,1H),2.68-2.50(m,1H),2.39-2.21(m,1H),1.35(t,J=7.1Hz,3H).

[0905] 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)

[0906]

[0907] The title compound was prepared from compound 11 (0.010 g, 0.028 mmol, 1.0 equiv) and tert-butyl 4-(2-aminoethyl)piperazine-1-carboxylate (20 mg, 0.09 mmol, 3 equiv) according to General Procedure 1. Preparative HPLC purification was accomplished as described in General Procedure 5 with a gradient elution from 10 to 45% CH3CN + 0.1% TFA / H2O + 0.1% TFA. Half of the resulting Boc intermediate was retained and deprotection of the remaining material was accomplished according to General Procedure 2 to afford the title compound 134 (3.6 mg, 0.0039 mmol, 28%) as a white solid. LC-MS: C 22 H 32 Calculated m / z for N8O3 = 456.3, found [M+H] + =457.3. 1H NMR(300MHz,MeOD)δ7.18(d,J=1.5Hz,1H),7.14-6.98(m,2H),5.06(s,2H),4.35(q,J=7.1Hz ,2H),4.25(s,2H),3.94(s,3H),3.32-3.14(m,6H),2.79-2.66(m,6H),1.35(t,J=7.1Hz,3H).

[0908] Example 1.48: 6-Amino-2-ethoxy-9-(2-methoxy-4-(((2-morpholinoethyl)amino)methyl)benzyl)-9H-purin-8-ol (Compound 135)

[0909]

[0910] 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 accomplished as described in General Procedure 5 with a gradient elution from 10% to 45% CH3CN + 0.1% TFA / H2O + 0.1% TFA to afford the title compound 135 (3.1 mg, 0.0039 mmol, 28%) as a white solid. LC-MS: C 22 H 31 Calculated m / z for N7O4 = 457.3, found [M+H] + =458.4. 1 H NMR (300MHz, MeOD) δ7.16(d,J=1.5Hz,1H),7.09(d,J=7.7Hz,1H),7.02(dd,J=7.7,1.6Hz,1H),5.05(s,2H),4.32(q,J=7.1Hz,2 H), 4.26 (s, 2H), 3.94 (s, 3H), 3.85-3.76 (m, 4H), 3.04 (t, J = 6.4Hz, 2H), 2.90-2.84 (m, 4H), 1.48 (s, 2H), 1.34 (t, J = 7.1Hz, 3H).

[0911] Example 1.49: 6-Amino-2-ethoxy-9-(4-(((2-hydroxy-3-morpholinopropyl)amino)methyl)-2-methoxybenzyl)-9H-purin-8-ol (Compound 136)

[0912]

[0913] The title compound was prepared from compound 11 (0.010 g, 0.028 mmol, 1.0 equiv) and 1-amino-3-(morpholin-4-yl)propan-2-ol (20 mg, 0.09 mmol, 3 equiv) according to General Procedure 1. Preparative HPLC purification was accomplished as described in General Procedure 5 with a gradient elution from 10% to 45% CH3CN + 0.1% TFA / H2O + 0.1% TFA to afford the title compound 136 (2.9 mg, 0.0036 mmol, 25%) as a white solid. LC-MS: C 23 H 33 Calculated m / z for N7O5 = 487.3, found [M+H] + =488.4. 1 H NMR(300MHz,MeOD)δ7.19(d,J=1.6Hz,1H),7.12-6.97(m,2H),5.05(s,2H),4.50-4.41(m,1H),4.34(q,J=7.1Hz,2H),4.27(s,2 H), 4.03-3.85 (m, 4H), 3.94 (s, 3H), 3.41-3.35 (m, 4H), 3.28-3.10 (m, 3H), 3.04 (dd, J = 12.9, 9.1Hz, 1H), 1.35 (t, J = 7.1Hz, 3H).

[0914] Example 1.50: 6-Amino-2-ethoxy-9-(2-methoxy-4-(((2-thiomorpholinoethyl)amino)methyl)-benzyl)-9H-purin-8-ol (Compound 137)

[0915]

[0916] The title compound was prepared from compound 11 (0.010 g, 0.028 mmol, 1.0 equiv) and 2-(thiomorpholin-4-yl)ethanamine (0.010 g, 0.09 mmol, 3 equiv) according to General Procedure 1. Preparative HPLC purification was accomplished as described in General Procedure 5 with a gradient elution from 10% to 45% CH3CN + 0.1% TFA / H2O + 0.1% TFA to afford the title compound 137 (3.5 mg, 0.0043 mmol, 31%) as a white solid. LC-MS: C 22 H 31 Calculated m / z for N7O3S = 473.2, found [M+H] + =473.3. 1H NMR(300MHz,MeOD)δ7.19-6.97(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=4.6Hz,4H),1.34(t,J=7.1Hz,3H).

[0917] Example 1.51: 9-(4-([4,4'-Bipiperidinyl]-1-ylmethyl)-2-methoxybenzyl)-6-amino-2-ethoxy-9H-purin-8-ol (Compound 138)

[0918]

[0919] The title compound was prepared from compound 11 (12 mg, 0.033 mmol, 1.0 equiv) and tert-butyl 4,4'-bipiperidine-1-carboxylate (35 mg, 0.13 mmol, 4.0 equiv) according to General Procedure 1. Preparative HPLC purification was accomplished as described in General Procedure 5 with a gradient elution from 10 to 45% CH3CN+0.1%TFA / H2O+0.1%TFA. Half of the resulting Boc intermediate was retained and deprotection of the remaining material was accomplished according to General Procedure 2 to afford the title compound 138 (8.0 mg, 0.0096 mmol, 58%) as a white solid. LC-MS: C 26 H 37 Calculated m / z for N7O3 = 495.3, found [M+H] + =496.4. 1 H NMR (300MHz, MeOD) δ7.16 (d, J = 1.5 Hz, 1H), 7.12-6.96 (m, 2H), 5.05 (s, 2H), 4.43-4.24 (m, 4H), 3.93 (s, 3H), 3.53 (d, J = 12. 1Hz, 2H), 3.43 (d, J = 12.6Hz, 2H), 2.95 (q, J = 10.1Hz, 4H), 2.00 (t, J = 12.4Hz, 4H), 1.58-1.44 (m, 6H), 1.33 (t, J = 7.1Hz, 3H).

[0920] Example 1.52: 9-(4-((3,9-diazaspiro[5.5]undec-3-yl)methyl)-2-methoxybenzyl)-6-amino-2-ethoxy-9H-purin-8-ol (Compound 139)

[0921]

[0922] The title compound was prepared from compound 11 (12 mg, 0.033 mmol, 1.0 equiv) and tert-butyl 3,9-diazaspiro[5.5]undecane-3-carboxylate (33 mg, 0.13 mmol, 4.0 equiv) according to General Procedure 1. Preparative HPLC purification was accomplished as described in General Procedure 5 with a gradient elution from 10 to 45% CH3CN+0.1%TFA / H2O+0.1%TFA. Half of the resulting Boc intermediate was retained and deprotection of the remaining material was accomplished according to General Procedure 2 to afford the title compound 139 (7.7 mg, 0.0093 mmol, 56%) as a white solid. LC-MS: C 25 H 35 Calculated m / z for N7O3 = 481.3, found [M+H] + =482.4. 1 H NMR(300MHz,MeOD)δ7.16(d,J=1.5Hz,1H),7.12-6.96(m,2H),5.05(s,2H),4.38-4.25(m,4H),3.94(s,3H),3.38(d ,J=12.7Hz,2H),3.24-3.18(m,6H),2.08-1.96(m,2H),1.96-1.90(m,2H),1.72-1.66(m,4H),1.33(t,J=7.1Hz,3H).

[0923] Example 1.53: 6-Amino-9-(4-((4-(2-aminoethyl)piperidin-1-yl)methyl)-2-methoxybenzyl)-2-ethoxy-9H-purin-8-ol (Compound 140)

[0924]

[0925] The title compound was prepared from compound 11 (30 mg, 0.083 mmol, 1.0 equiv) and tert-butyl N-[2-(piperidin-4-yl)ethyl]carbamate (94 mg, 0.41 mmol, 5.0 equiv) according to General Procedure 1. Preparative HPLC purification was accomplished as described in General Procedure 5 with a gradient elution from 10 to 45% CH3CN + 0.1% TFA / H2O + 0.1% TFA. Half of the resulting Boc intermediate was retained and deprotection of the remaining material was accomplished according to General Procedure 2 to afford the title compound 140 (7.7 mg, 0.0097 mmol, 23%) as a white solid. LC-MS: C 23 H 33 Calculated m / z for N7O3 = 455.3, found [M+H] + =456.3. 1H NMR (300MHz, MeOD) δ7.17(d,J=1.5Hz,1H),7.09(d,J=7.7Hz,1H),7.01(dd,J=7.7,1.5Hz,1H),5.06(s,2H),4.40-4.26(m,4H),3.94(s,3H) ,3.51(d,J=12.6Hz,2H),2.97(d,J=7.7Hz,5H),2.08-1.94(m,2H),1.64(dd,J=14.4,6.6Hz,2H),1.54-1.40(m,2H),1.34(t,J=7.1Hz,3H).

[0926] Example 1.54: 6-Amino-2-ethoxy-9-(4-(((4-(hydroxymethyl)benzyl)amino)methyl)-2-methoxybenzyl)-9H-purin-8-ol

[0927]

[0928] The title compound 12 was prepared from compound 11 (80 mg, 0.22 mmol, 1.0 equiv) and (4-(aminomethyl)phenyl)methanol (90 mg, 0.7 mmol, 3 equiv) according to General Procedure 1. Preparative HPLC purification was accomplished as described in General Procedure 5 with a gradient elution from 0% to 40% CH3CN + 0.1% TFA / H2O + 0.1% TFA to afford the title compound as a white solid (75 mg, 0.16 mmol, 73%). LC-MS: C 24 H 28 Calculated m / z for N6O4 = 464.2, found [M+H] + =465.3.

[0929] Example 1.55: 6-amino-9-(4-(((4-(chloromethyl)benzyl)amino)methyl)-2-methoxybenzyl)-2-ethoxy-9H-purin-8-ol

[0930]

[0931] The title compound 13 was prepared from compound 12 (75 mg, 0.16 mmol, 1.0 eq) and 10% SOCl2 / DCM (4 mL) according to General Procedure 3. The title compound was obtained as a yellow solid (65 mg, 0.14 mmol, 83%). LC-MS: C 24 H 27 Calculated m / z for ClN6O3 = 482.2, found [M+H] + =483.3.

[0932] Example 1.56: 6-Amino-9-(4-(((4-((dipropylamino)methyl)benzyl)amino)methyl)-2-methoxy-benzyl)-2-ethoxy-9H-purin-8-ol (Compound 141)

[0933]

[0934] The title compound was prepared from compound 13 (8.0 mg, 0.017 mmol, 1.0 equiv) and dipropylamine (11 μL, 0.083 mmol, 5.0 equiv) according to General Procedure 1. Preparative HPLC purification was accomplished as described in General Procedure 5 with a gradient elution from 10% to 45% CH 3 CN + 0.1% TFA / H 2 O + 0.1% TFA to afford the title compound 141 (2.7 mg, 0.0030 mmol, 18%) as a white solid. LC-MS: C 30 H 41 Calculated m / z for N7O3 = 547.3, found [M+H] + =548.3. 1 H NMR(300MHz,MeOD)δ7.63(s,4H),7.15(d,J=1.3Hz,1H),7.02(q,J=7.8Hz,2H),5.04(s,2H),4.42(s,2H),4.36 -4.22(m,6H),3.93(s,3H),3.15-2.98(m,4H),1.85-1.71(m,4H),1.32(t,J=7.1Hz,3H),0.99(t,J=7.3Hz,6H).

[0935] Example 1.57: 6-Amino-2-ethoxy-9-(2-methoxy-4-(((4-(morpholinomethyl)benzyl)amino)-methyl)benzyl)-9H-purin-8-ol (Compound 142)

[0936]

[0937] The title compound was prepared from compound 13 (8.0 mg, 0.017 mmol, 1.0 equiv) and morpholine (7.2 μL, 0.083 mmol, 5.0 equiv) according to General Procedure 1. Preparative HPLC purification was accomplished as described in General Procedure 5 with a gradient elution from 10% to 45% CH 3 CN + 0.1% TFA / H 2 O + 0.1% TFA to afford the title compound 142 (4.1 mg, 0.0047 mmol, 28%) as a white solid. LC-MS: C 28 H 35 The calculated m / z of N7O4 is 533.3, and the measured [M+H] +=534.3. 1 H NMR(300MHz,MeOD)δ7.62(s,4H),7.14(d,J=1.5Hz,1H),7.11-6.95(m,2H),5.04(s,2H) ,4.42(s,2H),4.39-4.23(m,6H),3.92(s,3H),4.14-3.67(m,8H),1.33(t,J=7.1Hz,3H).

[0938] 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)

[0939]

[0940] The title compound was prepared from compound 13 (8.0 mg, 0.017 mmol, 1.0 equiv) and tert-butyl N-(piperidin-4-yl)carbamate (17 mg, 0.083 mmol, 5.0 equiv) according to General Procedure 1. Preparative HPLC purification was accomplished as described in General Procedure 5 with a gradient elution from 10% to 55% CH3CN + 0.1% TFA / H2O + 0.1% TFA to afford the title compound 143 (8.6 mg, 0.0083 mmol, 49%) as a white solid. LC-MS: C 34 H 46 Calculated m / z for N8O5 = 646.4, found [M+H] + =647.5. 1 H NMR(300MHz,MeOD)δ7.61(s,4H),7.14(s,1H),7.09-6.95(m,2H),5.04(s,2H),4.39-4.23(m,8H),3.92(s,3H),3.63-3.5 7(m,1H),3.56-3.46(m,2H),3.18-3.04(m,2H),2.20-2.09(m,2H),1.77-1.63(m,2H),1.45(s,9H),1.33(t,J=7.1Hz,3H).

[0941] 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)

[0942]

[0943] Compound 143 (4.5 mg, 0.0045 mmol, 1.0 equiv) was deprotected according to General Procedure 2 to give the title compound 144 (4.3 mg, 0.0043 mmol, 96%) as a white solid. LC-MS: C 29 H 38 Calculated m / z for N8O3 = 546.3, found [M+H] + =547.4. 1 H NMR(300MHz,MeOD)δ7.60(s,4H),7.14(s,1H),7.02(q,J=7.8Hz,2H),5.04(s,2H),4.41-4.20(m,9H),3.92(s,3 H), 3.59 (d, J = 12.7Hz, 2H), 3.29-3.10 (m, 2H), 2.26 (d, J = 13.9Hz, 2H), 2.08-1.93 (m, 2H), 1.32 (t, J = 7.1Hz, 3H).

[0944] 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

[0945]

[0946] The title compound 14 was prepared from compound 11 (8 mg, 0.017 mmol, 1.0 equiv) and tert-butyl piperazine-1-carboxylate (15 mg, 0.083 mmol, 5.0 equiv) according to General Procedure 1. Preparative HPLC purification was accomplished as described in General Procedure 5 with a gradient elution from 10% to 55% CH3CN + 0.1% TFA / H2O + 0.1% TFA to afford the title compound (9.0 mg, 0.0092 mmol, 54%) as a white solid. LC-MS: C 33 H 44 Calculated m / z for N8O5 = 632.3, found [M+H] + =633.4. 1 H NMR(300MHz,MeOD)δ7.62(s,4H),7.17-7.11(m,1H),7.10-6.95(m,2H),5.04(s,2H),4.41(s,2H),4.3 8-4.23(m,6H),3.92(s,3H),3.72-3.66(m,4H),3.30-3.24(m,4H),1.49(s,9H),1.33(t,J=7.0Hz,3H).

[0947] 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)

[0948]

[0949] Compound 14 (5.0 mg, 0.0051 mmol, 1.0 equiv) was deprotected according to General Procedure 2 to give the title compound 145 (4.8 mg, 0.0049 mmol, 95%) as a white solid. LC-MS: C 28 H 36 The calculated m / z of N8O3 is 532.3, and the measured [M+H] + =533.4. 1 H NMR (300MHz, MeOD) δ7.49 (s, 4H), 7.16-6.95 (m, 3H), 5.04 (s, 2H), 4.32 (q, J = 7.1Hz, 2H), 4.26-4. 20(m,4H),3.92(s,3H),3.76(s,2H),3.33-3.24(m,4H),2.86-2.76(m,4H),1.32(t,J=6.7Hz,3H).

[0950] 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)

[0951]

[0952] The title compound was prepared from compound 13 (8.0 mg, 0.017 mmol, 1.0 equiv) and tert-butyl (4-(aminomethyl)benzyl)carbamate (20 mg, 0.083 mmol, 5.0 equiv) according to General Procedure 1. Preparative HPLC purification was accomplished as described in General Procedure 5 with a gradient elution from 10% to 55% CH3CN + 0.1% TFA / H2O + 0.1% TFA. Half of the resulting Boc intermediate was retained, and the remaining material was deprotected according to General Procedure 2 to afford the title compound 146 (5.4 mg, 0.0052 mmol, 61%) as a white solid. LC-MS: C 32 H 38 Calculated m / z for N8O3 = 582.3, found [M+H] + =583.4. 1H NMR(300MHz,MeOD)δ7.58(d,J=4.0Hz,8H),7.13(s,1H),7.07-6.94(m,2H),5.0 3(s,2H),4.36-4.21(m,10H),4.18(s,2H),3.92(s,3H),1.32(t,J=7.1Hz,3H).

[0953] Example 1.63: 6-Amino-9-(4-(((4-((Benzylamino)methyl)benzyl)amino)methyl)-2-methoxy-benzyl)-2-ethoxy-9H-purin-8-ol (Compound 147)

[0954]

[0955] The title compound was prepared from compound 13 (8.0 mg, 0.017 mmol, 1.0 equiv) and benzylamine (9.1 μL, 0.083 mmol, 5.0 equiv) according to General Procedure 1. Preparative HPLC purification was accomplished as described in General Procedure 5 with a gradient elution from 10% to 55% CH 3 CN + 0.1% TFA / H 2 O + 0.1% TFA to afford the title compound 147 (2.7 mg, 0.0030 mmol, 18%) as a white solid. LC-MS: C 31 H 35 Calculated m / z for N7O3 = 553.3, found [M+H] + =554.4. 1 H NMR(300MHz,MeOD)δ7.59(s,4H),7.49(s,5H),7.13(d,J=1.3Hz,1H),7.08-6.9 4(m,2H),5.03(s,2H),4.37-4.22(m,10H),3.92(s,3H),1.32(t,J=7.1Hz,3H).

[0956] 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)

[0957]

[0958] To a solution of iminodiacetic acid (14 mg, 0.10 mmol, 5.0 equiv) in aqueous NaHCO3 (0.272 mL, 1.14 M, 15.0 equiv) was added a solution of compound 11 (0.010 g, 0.021 mmol, 1.0 equiv) in 100 μL DMF. The resulting solution was heated to 60°C and maintained for 18 h. Preparative HPLC purification was accomplished as described in General Procedure 5 with a gradient elution of 10% to 45% CH3CN + 0.1% TFA / H2O + 0.1% TFA to afford the title compound 151 (1.5 mg, 0.0026 mmol, 26%) as a white solid. LC-MS: C 28 H 33 Calculated m / z for N7O7 = 579.2, found [M+H] + =580.3. 1 H NMR(300MHz,MeOD)δ7.63-7.47(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=7.1Hz,3H).

[0959] 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)

[0960]

[0961] 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-dioxopyrrolidin-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, after which L-cysteine ​​(12 mg, 0.096 mmol, 2.0 equiv) was added and the resulting solution was stirred at room temperature for 1 h. HO (200 μL) was then added and the reaction mixture was stirred at room temperature for an additional 2 h. Preparative HPLC purification was accomplished as described in General Procedure 5 with a gradient elution of 5% to 45% CH 3 CN + 0.1% TFA / H 2 O + 0.1% TFA to afford the title compound 148 (5.1 mg, 0.0047 mmol, 9.8%) as a white solid. LC-MS: C33 H 47 Calculated m / z for N9O9S = 745.3, found [M+H] + =746.4.

[0962] 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-azaeicosane-20-oic acid (Compound 149)

[0963]

[0964] To a solution of compound 100 (0.020 g, 0.048 mmol, 1.0 equiv) in DMF (500 μL) was added compound 56 (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, after which L-cysteine ​​(12 mg, 0.096 mmol, 2.0 equiv) was added and the resulting solution was stirred at room temperature for 1 h. H2O (200 μL) was then added and the reaction mixture was stirred at room temperature for an additional 2 h. Preparative HPLC purification was accomplished as described in General Procedure 5 with a gradient elution of 5% to 45% CH3CN+0.1%TFA / H2O+0.1%TFA to afford the title compound 149 (0.5 mg, 0.0004 mmol, 1%) as a white solid. LC-MS: C 36 H 53 N9O 12 Calculated m / z for S = 835.4, found [M+H] + =836.5.

[0965] Example 1.67: Methyl 4-((6-amino-2-ethoxy-8-methoxy-9H-purin-9-yl)methyl)benzoate

[0966]

[0967] To a solution of compound 8 (0.75 g, 1.7 mmol, 1.0 equiv) and methyl 4-(bromomethyl)benzoate (0.39 g, 1.7 mmol, 1.0 equiv) in DMF (8 mL) was added CsCO (0.56 g, 1.7 mmol, 1.0 equiv). The resulting suspension was stirred at room temperature for 18 h and then concentrated in vacuo. Reverse phase rapid purification was accomplished using a 60 g C18 column with a gradient elution of 10% to 50% CHCN + 0.1% TFA / H2O + 0.1% TFA as described in General Procedure 5. The title compound 15 (0.65 g, 1.4 mmol, 81%) was obtained as a white solid. LC-MS: C 17 H 19 Calculated m / z for N5O4 = 357.1, found [M+H] + =358.2.

[0968] Example 1.68: (4-((6-amino-2-ethoxy-8-methoxy-9H-purin-9-yl)methyl)phenyl)methanol

[0969]

[0970] Lithium aluminum hydride (64 mg, 1.7 mmol, 3.0 equiv) was added in small portions to a stirred solution of compound 15 (0.20 g, 0.56 mmol, 1.0 equiv) in anhydrous THF (15 mL) cooled to 0°C over 5 min. The resulting suspension was allowed to warm to room temperature and stirred for 15 min before being quenched with H2O (10 mL) and diluted with MeOH (50 mL) and filtered through a plug of celite. Reverse phase flash purification was accomplished as described in General Procedure 5 using a 60 g C18 column eluting with a gradient of 20% to 65% CH3CN+0.1%TFA / H2O+0.1%TFA to afford the title product 16 (0.12 g, 0.36 mmol, 65%) as a white solid. LC-MS: C 16 H 19 Calculated m / z for N5O3 = 329.2, found [M+H] + =330.2.

[0971] Example 1.69: 6-amino-9-(3-(chloromethyl)benzyl)-2-ethoxy-9H-purin-8-ol

[0972]

[0973] The title compound 17 was prepared from compound 16 (0.10 g, 0.30 mmol, 1.0 eq) and 10% SOCl2 / DCM (5 mL) according to General Procedure 3. The title compound was obtained as a yellow solid (assuming quantitative yield). LC-MS: C 15 H 16 Calculated m / z for ClN5O2 = 333.1, found [M+H] + =333.2.

[0974] Example 1.70: 6-Amino-2-ethoxy-9-(4-(piperazin-1-ylmethyl)benzyl)-9H-purin-8-ol (Compound 150)

[0975]

[0976] 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 accomplished as described in General Procedure 5 with a gradient elution from 10 to 45% CH3CN + 0.1% TFA / H2O + 0.1% TFA. Half of the resulting Boc intermediate was retained and deprotection of the remaining material was accomplished according to General Procedure 2 to afford the title compound 150 (5.1 mg, 0.0070 mmol, 30%) as a white solid. LC-MS: C 19 H 25 Calculated m / z for N7O2 = 383.2, found [M+H] + =383.3. 1 H NMR(300MHz,MeOD)δ7.45(d,J=8.1Hz,2H),7.37(d,J=8.1Hz,2H),5.02(s,2H),4.39(q, J=7.1Hz,2H),3.80(s,2H),3.31-3.25(m,4H),2.91-2.82(m,4H),1.38(t,J=7.1Hz,3H).

[0977] Example 1.71: Methyl 4-((6-amino-2-ethoxy-8-methoxy-9H-purin-9-yl)methyl)-3-fluorobenzoate

[0978]

[0979] To 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 CsCO (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 rapid purification was accomplished using a 60 g C18 column with a gradient elution from 10% to 50% CHCN+0.1% TFA / H0+0.1% TFA as described in General Procedure 5. The title compound 18 (0.50 g, 1.3 mmol, 66%) was obtained as a white solid. LC-MS: C 17 H 18 Calculated m / z for FN5O4 = 375.1, found [M+H] + =376.2.

[0980] Example 1.72: (4-((6-amino-2-ethoxy-8-methoxy-9H-purin-9-yl)methyl)-3-fluorophenyl)-methanol

[0981]

[0982] To a stirred solution of compound 18 (0.50 g, 1.3 mmol, 1.0 equiv) in anhydrous THF (4 mL) cooled to 0°C was added lithium aluminum hydride (51 mg, 1.3 mmol, 1.0 equiv). The resulting suspension was allowed to warm to room temperature and stirred for 15 min before being diluted with MeOH (50 mL) and filtered through a plug of celite. Reverse phase flash purification was accomplished as described in General Procedure 5 using a 60 g C18 column eluting with a gradient of 10% to 40% CH3CN+0.1%TFA / H2O+0.1%TFA to afford the title product 19 (0.40 g, 1.2 mmol, 86%) as a white solid. LC-MS: C 16 H 18 Calculated m / z for FN5O3 = 347.1, found [M+H] + =348.2.

[0983] Example 1.73: 6-amino-9-(4-(chloromethyl)-2-fluorobenzyl)-2-ethoxy-9H-purin-8-ol

[0984]

[0985] The title compound 20 was prepared from compound 19 (0.20 g, 0.58 mmol, 1.0 eq) and 10% SOCl2 / DCM (5 mL) according to General Procedure 3. The title compound was obtained as a yellow solid (assuming quantitative yield). LC-MS: C 15 H 15 Calculated m / z for ClFN5O2 = 351.1, found [M+H] + =352.2.

[0986] Example 1.74: 6-Amino-2-ethoxy-9-(2-fluoro-4-(piperazin-1-ylmethyl)benzyl)-9H-purin-8-ol (Compound 152)

[0987]

[0988] The title compound was prepared from compound 20 (0.020 g, 0.057 mmol, 1.0 equiv) and tert-butyl piperazine-1-carboxylate (32 mg, 0.17 mmol, 3.0 equiv) according to General Procedure 1. Preparative HPLC purification was accomplished as described in General Procedure 5 with a gradient elution from 10 to 45% CH3CN + 0.1% TFA / H2O + 0.1% TFA. Half of the resulting Boc intermediate was retained and deprotection of the remaining material was accomplished according to General Procedure 2 to afford the title compound 152 (12 mg, 0.016 mmol, 57%) as a white solid. LC-MS: C 19 H 24 Calculated m / z for FN7O2 = 401.2, found [M+H] + =402.3. 1 H NMR(300MHz,MeOD)δ7.34(t,J=7.7Hz,1H),7.28-7.14(m,2H),5.10(s,2H),4.41(q,J=7.1 Hz, 2H), 3.80 (s, 2H), 3.30 (d, J = 5.3Hz, 4H), 2.88 (t, J = 5.3Hz, 4H), 1.38 (t, J = 7.1Hz, 3H).

[0989] Example 1.75: 6-Amino-9-(4-((4-aminopiperidin-1-yl)methyl)-2-fluorobenzyl)-2-ethoxy-9H-purin-8-ol (Compound 153)

[0990]

[0991] The title compound was prepared from compound 20 (0.020 g, 0.057 mmol, 1.0 equiv) and tert-butyl N-(4-aminobutyl)carbamate (34 mg, 0.17 mmol, 3.0 equiv) according to General Procedure 1. Preparative HPLC purification was accomplished as described in General Procedure 5 with a gradient elution from 10 to 45% CH3CN + 0.1% TFA / H2O + 0.1% TFA. Half of the resulting Boc intermediate was retained and deprotection of the remaining material was accomplished according to General Procedure 2 to afford the title compound 153 (0.010 g, 0.013 mmol, 46%) as a white solid. LC-MS: C 19 H 24 Calculated m / z for FN7O2 = 415.2, found [M+H] + =416.3. 1 H NMR(300MHz,MeOD)δ7.48-7.27(m,3H),5.13(s,2H),4.43-4.30(m,4H),3.65-3.54(m,2H),3.49- 3.39(m,1H),3.16(t,J=12.6Hz,2H),2.32-2.21(m,2H),2.05-1.88(m,2H),1.36(t,J=7.1Hz,3H).

[0992] Example 1.76: 2-Butoxy-9-(tetrahydro-2H-pyran-2-yl)-9H-purin-6-amine

[0993]

[0994] To a solution of compound 2 (9.8 g, 39 mmol, 1.0 equiv) in nBuOH (100 mL) was added KOtBu (8.7 g, 77 mmol, 2.0 equiv). The resulting solution was stirred at 100 ° C for 18 h, after which the solvent was removed in vacuo. The resulting solid was dissolved in EtOH (100 mL) and H2O (20 mL), then extracted with EtOAc (2 x 50 mL). The pooled organics were washed with brine (5 mL), then dried over MgSO4 and concentrated in vacuo to give the title compound 21 as an orange solid (assuming quantitative yield). LC-MS: C 14 H 21 Calculated m / z for N5O2 = 291.2, found [M+H] + =292.2.

[0995] Example 1.77: 8-Bromo-2-butoxy-9-(tetrahydro-2H-pyran-2-yl)-9H-purin-6-amine

[0996]

[0997] To a suspension of compound 21 (11 g, 39 mmol, 1.0 equiv) in DCM (50 mL) was added NBS (6.9 g, 39 mmol, 1.0 equiv). The resulting mixture was stirred at room temperature for 18 h, after which the reaction was quenched by the addition of 1 M NaHSO (50 mL) and the reaction was rapidly stirred for 30 min. The resulting mixture was diluted with DCM (100 mL) and the separated organics were washed with 1 M NaHSO (2×50 mL). The pooled organics were dried over MgSO, filtered, and then concentrated in vacuo to give the title compound 22 (13 g, 34 mmol, 88%) as an orange solid. LC-MS: C 14 H 20 Calculated m / z for BrN5O2 = 369.1, found [M+H] + =370.2.

[0998] Example 1.78: 2-Butoxy-8-methoxy-9-(tetrahydro-2H-pyran-2-yl)-9H-purin-6-amine

[0999]

[1000] To a solution of compound 22 (13 g, 34 mmol, 1.0 equiv) in MeOH (100 mL) was added NaOMe (5.47 g, 101 mmol, 3.00 equiv). The resulting mixture was stirred at room temperature for 72 h. NaOMe (3.6 g, 66 mmol, 1.0 equiv) was further added and the mixture was heated to reflux and maintained for 72 h. The solvent was removed in vacuo and the crude material was redissolved in EtOAc (250 mL) and then washed with 1 M NaH2PO4 (150 mL). The aqueous layer was then extracted with EtOAc (3×150 mL), the pooled organic layers were dried over MgSO4, and the solution was concentrated in vacuo to give the title compound 23 (11 g, 34 mmol, 100%) as a red solid. LC-MS: C 15 H 23 Calculated m / z for N5O3 = 321.2, found [M+H] + =322.3.

[1001] Example 1.79: 2-Butoxy-8-methoxy-9-(tetrahydro-2H-pyran-2-yl)-9H-purin-6-amine

[1002]

[1003] Compound 23 (11 g, 34 mmol, 1.0 equiv) was dissolved in 10% TFA / MeOH (50 mL) and the resulting solution was stirred at room temperature for 2 weeks. The solvent was removed in vacuo and the residue was co-evaporated with MeOH (2×20 mL) to give the title compound 24 (9.8 g, 28 mmol, 83%) as an orange solid. LC-MS: C 10 H 15 Calculated m / z for N5O2 = 237.1, found [M+H] + =238.2.

[1004] Example 1.80: Methyl 4-((6-amino-2-butoxy-8-methoxy-9H-purin-9-yl)methyl)-3-methoxybenzoate

[1005]

[1006] To a solution of compound 24 (570 mg, 2.4 mmol, 1.0 equiv) and methyl 4-(bromomethyl)-3-methoxybenzoate (620 mg, 2.4 mmol, 1.0 equiv) in DMF (5 mL) was added CsCO (620 mg, 2.4 mmol, 1.0 equiv). The resulting suspension was stirred at room temperature for 18 h and then concentrated in vacuo. Reverse phase rapid purification was accomplished using a 60 g C18 column with a gradient elution from 10% to 60% CHCN + 0.1% TFA / H2O + 0.1% TFA as described in General Procedure 5. The title compound 25 (240 mg, 0.58 mmol, 24%) was obtained as a white solid. LC-MS: C 20 H 25 Calculated m / z for N5O5 = 415.2, found [M+H] + =416.3.

[1007] Example 1.81: (4-((6-amino-2-butoxy-8-methoxy-9H-purin-9-yl)methyl)-3-methoxyphenyl)-methanol

[1008]

[1009] To a stirred solution of compound 25 (240 mg, 0.58 mmol, 1.0 eq.) in anhydrous THF (5 mL) cooled to 0°C was added lithium aluminum hydride (22 mg, 0.58 mmol, 1.0 eq.). The resulting suspension was allowed to warm to room temperature and stirred for 15 min, after which it was diluted with MeOH (25 mL) and filtered through a plug of celite. The filtrate was concentrated in vacuo to afford the title compound 26 (220 mg, 0.57 mmol, 98%). LC-MS: C 19H 25 Calculated m / z for N5O4 = 387.2, found [M+H] + =388.3.

[1010] Example 1.82: 6-Amino-2-butoxy-9-(4-(chloromethyl)-2-methoxybenzyl)-9H-purin-8-ol

[1011]

[1012] The title compound 27 was prepared from compound 26 (220 mg, 0.57 mmol, 1.0 eq) and 15% SOCl2 / DCM (5 mL) according to General Procedure 3. The title compound was obtained as a yellow solid (210 mg, 0.577 mmol, 98.8%). LC-MS: C 18 H 22 Calculated m / z for ClN5O3 = 391.1, found [M+H] + =392.2.

[1013] Example 1.83: 6-Amino-2-butoxy-9-(2-methoxy-4-(piperazin-1-ylmethyl)benzyl)-9H-purin-8-ol (Compound 154)

[1014]

[1015] The title compound was prepared from compound 27 (28 mg, 0.071 mmol, 1.0 equiv) and tert-butyl piperazine-1-carboxylate (53 mg, 0.28 mmol, 4.0 equiv) according to General Procedure 1. Preparative HPLC purification was accomplished as described in General Procedure 5 with a gradient elution from 10% to 55% CH3CN + 0.1% TFA / H2O + 0.1% TFA. Half of the resulting Boc intermediate was retained and deprotection of the remaining material was accomplished according to General Procedure 2 to afford the title compound 154 (6.8 mg, 0.0087 mmol, 24%) as a white solid. LC-MS: C 22 H 31 Calculated m / z for N7O3 = 441.2, found [M+H] + =442.3. 1H NMR(300MHz,MeOD)δ7.07-6.94(m,2H),6.90(d,J=7.7Hz,1H),5.03(s,2H),4.28(t,J=6.6Hz,2H),3.90(s,3H),3 .76(s,2H),3.34-3.24(m,4H),2.84(s,4H),1.72(p,J=6.7Hz,2H),1.46(h,J=7.4Hz,2H),0.97(t,J=7.4Hz,3H).

[1016] Example 1.84: 6-Amino-9-(4-((4-aminopiperidin-1-yl)methyl)-2-methoxybenzyl)-2-butoxy-9H-purin-8-ol (Compound 155)

[1017]

[1018] The title compound was prepared from compound 27 (25 mg, 0.064 mmol, 1.0 equiv) and tert-butyl N-(4-aminobutyl)carbamate (51 mg, 0.26 mmol, 4.0 equiv) according to General Procedure 1. Preparative HPLC purification was accomplished as described in General Procedure 5 with a gradient elution from 10 to 45% CH3CN + 0.1% TFA / H2O + 0.1% TFA. Deprotection was accomplished according to General Procedure 2 to afford the title compound 155 (6.1 mg, 0.0076 mmol, 24%) as a white solid. LC-MS: C 23 H 33 Calculated m / z for N7O3 = 455.3, found [M+H] + =456.3. 1 H NMR(300MHz,MeOD)δ7.15(s,1H),7.03(q,J=7.9Hz,2H),5.05(s,2H),4.35-4.20(m,4H),3.94(s,3H),3.61(d,J=12.9Hz,2H),3.45(s,1H) ,3.14(t,J=13.1Hz,2H),2.27(d,J=14.7Hz,2H),1.93(d,J=13.3Hz,2H),1.78-1.63(m,2H),1.46(h,J=7.4Hz,2H),0.98(t,J=7.4Hz,2H).

[1019] Example 1.85: 6-Amino-9-(4-((4-(aminomethyl)piperidin-1-yl)methyl)-2-methoxybenzyl)-2-butoxy-9H-purin-8-ol (Compound 156)

[1020]

[1021] The title compound was prepared from compound 27 (30 mg, 0.077 mmol, 1.0 equiv) and tert-butyl N-(piperidin-4-ylmethyl)carbamate (49 mg, 0.23 mmol, 3.0 equiv) according to General Procedure 1. Preparative HPLC purification was accomplished as described in General Procedure 5 with a gradient elution from 10% to 55% CH3CN + 0.1% TFA / H2O + 0.1% TFA. Deprotection was accomplished according to General Procedure 2 to afford the title compound 156 (8.0 mg, 0.0099 mmol, 13%) as a white solid. LC-MS: C 24 H 35 Calculated m / z for N7O3 = 469.3, found [M+H] + =470.4. 1 H NMR(300MHz,MeOD)δ7.16(s,1H),7.10-6.96(m,2H),5.05(s,2H),4.34-4.20(m,4H),3.94(s,3H),3.62-3.56(m,2H), 3.12-2.96(m,2H),2.95-2.87(m,2H),2.14-1.87(m,4H),1.79-1.63(m,2H),1.60-1.35(m,3H),0.96(t,J=7.4Hz,3H).

[1022] Example 1.86: 9-(4-((3,9-diazaspiro[5.5]undec-3-yl)methyl)-2-methoxybenzyl)-6-amino-2-butoxy-9H-purin-8-ol (Compound 157)

[1023]

[1024] 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 accomplished as described in General Procedure 5 with a gradient elution from 10% to 55% CH3CN+0.1%TFA / H2O+0.1%TFA. Deprotection was accomplished according to General Procedure 2 to afford the title compound 157 (0.010 g, 0.012 mmol, 15%) as a white solid. LC-MS: C 27 H 39 Calculated m / z for N7O3 = 509.3, found [M+H] + =510.3. 1HNMR(300MHz,MeOD)δ7.15(s,1H),7.03(q,J=7.7Hz,2H),5.05(s,2H),4.32(s,2H),4.25(t,J=6.6Hz,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=7.4Hz,3H).

[1025] Example 1.87: 6-Amino-9-(4-((4-(2-aminoethyl)piperidin-1-yl)methyl)-2-methoxybenzyl)-2-butoxy-9H-purin-8-ol (Compound 158)

[1026]

[1027] The title compound was prepared from compound 27 (30 mg, 0.077 mmol, 1.0 equiv) and tert-butyl N-[2-(piperidin-4-yl)ethyl]carbamate (52 mg, 0.23 mmol, 3.0 equiv) according to General Procedure 1. Preparative HPLC purification was accomplished as described in General Procedure 5 with a gradient elution from 10% to 55% CH3CN + 0.1% TFA / H2O + 0.1% TFA. Deprotection was accomplished according to General Procedure 2 to afford the title compound 158 (9.0 mg, 0.011 mmol, 14%) as a white solid. LC-MS: C 25 H 37 Calculated m / z for N7O3 = 483.3, found [M+H] + =484.3. 1 HNMR(300MHz,MeOD)δ7.15(s,1H),7.02(q,J=7.9Hz,2H),5.05(s,2H),4.32-4.20(m,4H),3.95(s,3H),3.50 (s,2H),3.05-2.93(m,6H),2.06-1.96(m,2H),1.78-1.60(m,4H),1.53-1.35(m,3H),0.96(t,J=7.4Hz,3H).

[1028] Example 1.88: 6-Amino-2-butoxy-9-(4-(((2-hydroxy-2-methylpropyl)amino)methyl)-2-methoxybenzyl)-9H-purin-8-ol (Compound 159)

[1029]

[1030] The title compound was prepared from compound 27 (30 mg, 0.077 mmol, 1.0 equiv) and 1-amino-2-methylpropan-2-ol (0.020 g, 0.23 mmol, 3.0 equiv) according to General Procedure 1. Preparative HPLC purification was accomplished as described in General Procedure 5 with a gradient elution from 10% to 45% CH3CN + 0.1% TFA / H2O + 0.1% TFA to afford the title compound 159 (0.010 g, 0.030 mmol, 39%) as a white solid. LC-MS: C 22 H 32 Calculated m / z for N6O4 = 444.2, found [M+H] + =445.3. 1 H NMR(300MHz,MeOD)δ7.18(s,1H),7.08-6.96(m,2H),5.04(s,2H),4.30-4.19(m,4H),3.94(s,3H) ),2.93-2.83(m,2H),1.78-1.63(m,2H),1.55-1.33(m,2H),1.29(s,6H),0.96(t,J=7.4Hz,3H).

[1031] Example 1.89: 6-Amino-9-(4-(((4-aminobutyl)amino)methyl)-2-methoxybenzyl)-2-butoxy-9H-purin-8-ol (Compound 160)

[1032]

[1033] The title compound was prepared from compound 27 (30 mg, 0.077 mmol, 1.0 equiv) and tert-butyl N-(4-aminobutyl)carbamate (43 mg, 0.23 mmol, 3.0 equiv) according to General Procedure 1. Preparative HPLC purification was accomplished as described in General Procedure 5 with a gradient elution from 10% to 55% CH3CN + 0.1% TFA / H2O + 0.1% TFA. Deprotection was accomplished according to General Procedure 2 to afford the title compound 160 (0.010 g, 0.013 mmol, 17%) as a white solid. LC-MS: C 22 H 33 The calculated m / z of N7O3 is 443.3, and the measured [M+H] + =444.3. 1H NMR(300MHz,MeOD)δ7.14(s,1H),7.08-6.94(m,2H),5.04(s,2H),4.30-4.17(m,4H),3.94(s,3H),3.1 0(t,J=7.6Hz,2H),2.98(t,J=7.3Hz,2H),1.89-1.62(m,8H),1.55-1.37(m,2H),0.97(t,J=7.4Hz,3H).

[1034] 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)

[1035]

[1036] The title compound was prepared from compound 27 (30 mg, 0.077 mmol, 1.0 equiv) and (2R,3S)-2-aminopentane-1,3-diol (27 mg, 0.23 mmol, 3.0 equiv) according to General Procedure 1. Preparative HPLC purification was accomplished as described in General Procedure 5 with a gradient elution from 10% to 45% CH3CN + 0.1% TFA / H2O + 0.1% TFA to afford the title compound 161 (1.9 mg, 0.0027 mmol, 3.5%) as a white solid. LC-MS: C 24 H 36 Calculated m / z for N6O4 = 472.3, found [M+H] + =473.4. 1 H NMR (300MHz, MeOD) δ7.19(s,1H),7.04(s,2H),5.05(s,2H),4.42-4.19(m,4H),3.94(s,3H),3.88(dd,J=12.1,3.9Hz,1H),3.76(dd,J=1 2.1,7.2Hz,1H),3.15(dt,J=7.3,4.2Hz,1H),1.88(s,1H),1.78-1.63(m,2H),1.56-1.38(m,3H),1.38-1.25(m,1H),1.04-0.87(m,9H).

[1037] Example 1.91: 6-Amino-9-(4-(aminomethyl)-2-methoxybenzyl)-2-butoxy-9H-purin-8-ol (Compound 162)

[1038]

[1039] The title compound was prepared from compound 27 (13 mg, 0.033 mmol, 1.0 equiv) and ammonia / DMF (83 μL, 2 M, 5.0 equiv) according to General Procedure 1. Preparative HPLC purification was accomplished as described in General Procedure 5 with a gradient elution from 10% to 45% CH 3 CN + 0.1% TFA / H 2 O + 0.1% TFA to afford the title compound 162 (1.6 mg, 0.0027 mmol, 8.1%) as a white solid. LC-MS: C 18 H 24 Calculated m / z for N6O3 = 372.2, found [M+H] + =373.3. 1 H NMR(300MHz,MeOD)δ7.10(d,J=1.6Hz,1H),7.03(d,J=7.8Hz,1H),6.95(dd,J=7.7,1.6Hz,1H),5.04(s,2H),4 .25(t,J=6.5Hz,2H),4.10(s,2H),3.93(s,3H),1.79-1.63(m,2H),1.58-1.37(m,2H),0.97(t,J=7.4Hz,3H).

[1040] Example 1.92: 6-Amino-2-butoxy-9-(2-methoxy-4-((methylamino)methyl)benzyl)-9H-purin-8-ol (Compound 163)

[1041]

[1042] The title compound was prepared from compound 27 (13 mg, 0.033 mmol, 1.0 equiv) and methylamine hydrochloride (11.2 mg, 0.17 mmol, 5.0 equiv) according to General Procedure 1. Preparative HPLC purification was accomplished as described in General Procedure 5 with a gradient elution from 10% to 45% CH3CN + 0.1% TFA / H2O + 0.1% TFA to afford the title compound 163 (2.6 mg, 0.0042 mmol, 13%) as a white solid. LC-MS: C 19 H 26 The calculated m / z of N6O3 is 386.2, and the measured [M+H] + =387.3. 1H NMR(300MHz,MeOD)δ7.11(s,1H),7.08-6.93(m,2H),5.04(s,2H),4.24(t,J=6.5Hz,2H),4.17(s,2H), 3.94 (s, 3H), 2.73 (s, 3H), 1.70 (p, J = 6.7Hz, 2H), 1.46 (dq, J = 14.5, 7.3Hz, 2H), 0.96 (t, J = 7.4Hz, 3H).

[1043] Example 1.93: 6-Amino-2-butoxy-9-(4-((cyclobutylamino)methyl)-2-methoxybenzyl)-9H-purin-8-ol (Compound 164)

[1044]

[1045] The title compound was prepared from compound 27 (13 mg, 0.033 mmol, 1.0 equiv) and cyclobutylamine (14 μL, 0.17 mmol, 5.0 equiv) according to General Procedure 1. Preparative HPLC purification was accomplished as described in General Procedure 5 with a gradient elution from 10% to 45% CH 3 CN + 0.1% TFA / H 2 O + 0.1% TFA to afford the title compound 164 (5.3 mg, 0.0081 mmol, 25%) as a white solid. LC-MS: C 22 H 30 Calculated m / z for N6O3 = 426.2, found [M+H] + =427.3. 1 H NMR(300MHz,MeOD)δ7.19-6.92(m,3H),5.05(s,2H),4.26(t,J=6.5Hz,2H),4.07(s,2H),3.94(s,3H),3.80(p,J =8.0Hz,1H),2.50-2.02(m,4H),2.03-1.77(m,2H),1.78-1.61(m,2H),1.60-1.37(m,2H),0.97(t,J=7.4Hz,3H).

[1046] Example 1.94: 6-Amino-9-(4-((Benzylamino)methyl)-2-methoxybenzyl)-2-butoxy-9H-purin-8-ol (Compound 165)

[1047]

[1048] The title compound was prepared from compound 27 (13 mg, 0.033 mmol, 1.0 equiv) and benzylamine (18 μL, 0.17 mmol, 5.0 equiv) according to General Procedure 1. Preparative HPLC purification was accomplished as described in General Procedure 5 with a gradient elution from 10% to 45% CH 3 CN + 0.1% TFA / H 2 O + 0.1% TFA to afford the title compound 165 (7.8 mg, 0.011 mmol, 34%) as a white solid. LC-MS: C 25 H 30 Calculated m / z for N6O3 = 462.2, found [M+H] + =463.3. 1 H NMR (300MHz, MeOD) δ7.48 (s, 5H), 7.13 (d, J = 1.5Hz, 1H), 7.10-6.94 (m, 2H), 5.04 (s, 2H), 4. 32-4.16(m,6H),3.93(s,3H),1.86-1.62(m,2H),1.57-1.36(m,2H),0.96(t,J=7.4Hz,3H).

[1049] Example 1.95: Methyl 4-((6-amino-2-butoxy-8-methoxy-9H-purin-9-yl)methyl)benzoate

[1050]

[1051] To a solution of compound 24 (0.90 g, 3.8 mmol, 1.0 equiv) and methyl 4-(bromomethyl)-3-methoxybenzoate (0.87 g, 3.8 mmol, 1.0 equiv) in DMF (8 mL) was added CsCO (1.3 g, 3.8 mmol, 1.0 equiv). The resulting suspension was stirred at room temperature for 18 h and then concentrated in vacuo. Reverse phase rapid purification was accomplished using a 60 g C18 column with a gradient elution from 10% to 60% CHCN + 0.1% TFA / H2O + 0.1% TFA as described in General Procedure 5. The title compound 28 (0.23 g, 0.60 mmol, 16%) was obtained as a white solid. LC-MS: C 19 H 23 Calculated m / z for N5O4 = 385.2, found [M+H] + =386.3.

[1052] Example 1.96: (4-((6-amino-2-butoxy-8-methoxy-9H-purin-9-yl)methyl)phenyl)methanol

[1053]

[1054] To a stirred solution of compound 28 (0.23 g, 0.60 mmol, 1.0 equiv) in anhydrous THF (8 mL) cooled to 0°C was added lithium aluminum hydride (23 mg, 0.60 mmol, 1.0 equiv). The resulting suspension was stirred for 5 min, then allowed to warm to room temperature and stirred for an additional 15 min before being diluted with MeOH (50 mL) and filtered through a plug of celite. The filtrate was concentrated in vacuo to give the title compound 29 (0.20 g, 0.56 mmol, 94%) as a yellow solid. LC-MS: C 18 H 23 Calculated m / z for N5O3 = 357.2, found [M+H] + =358.3.

[1055] Example 1.97: 6-Amino-2-butoxy-9-(4-(chloromethyl)benzyl)-9H-purin-8-ol

[1056]

[1057] The title compound 30 was prepared from compound 29 (200 mg, 0.56 mmol, 1.0 eq) and 10% SOCl2 / DCM (10 mL) according to General Procedure 3. The title compound was obtained as a yellow solid (assuming quantitative yield). LC-MS: C 17 H 20 Calculated m / z for ClN5O2 = 361.1, found [M+H] + =396.2.

[1058] Example 1.98: 6-Amino-2-butoxy-9-(4-(piperazin-1-ylmethyl)benzyl)-9H-purin-8-ol (Compound 166)

[1059]

[1060] The title compound was prepared from compound 30 (30 mg, 0.083 mmol, 1.0 equiv) and tert-butyl piperazine-1-carboxylate (77 mg, 0.42 mmol, 5.0 equiv) according to General Procedure 1. Preparative HPLC purification was accomplished as described in General Procedure 5 with a gradient elution from 10% to 55% CH3CN + 0.1% TFA / H2O + 0.1% TFA. Half of the resulting Boc intermediate was retained and deprotection of the remaining material was accomplished according to General Procedure 2 to afford the title compound 166 (0.010 g, 0.014 mmol, 33%) as a white solid. LC-MS: C 21 H 29 Calculated m / z for N7O2 = 411.2, found [M+H]+ =412.3. 1 H NMR(300MHz,MeOD)δ7.07-6.94(m,2H),6.90(d,J=7.7Hz,1H),5.03(s,2H),4.28(t,J=6.6Hz,2H),3.90(s,3H),3 .76(s,2H),3.34-3.24(m,4H),2.84(s,4H),1.72(p,J=6.7Hz,2H),1.46(h,J=7.4Hz,2H),0.97(t,J=7.4Hz,3H).

[1061] Example 1.99: 2-Butoxy-9-((6-chloropyridin-3-yl)methyl)-8-methoxy-9H-purin-6-amine

[1062]

[1063] To a solution of compound 24 (0.70 g, 3.0 mmol, 1.0 equiv) and 2-chloro-5-(chloromethyl)pyridine (0.48 g, 3.0 mmol, 1.0 equiv) in DMF (5 mL) was added CsCO (0.96 g, 3.0 mmol, 1.0 equiv). The resulting suspension was stirred at room temperature for 18 h and then concentrated in vacuo. Reverse phase rapid purification was accomplished using a 60 g C18 column with a gradient elution from 10% to 60% CHCN + 0.1% TFA / H2O + 0.1% TFA as described in General Procedure 5. The title compound 31 (0.65 g, 1.8 mmol, 61%) was obtained as a white solid. LC-MS: C 16 H 19 Calculated m / z for ClN6O2 = 362.1, found [M+H] + =363.2.

[1064] Example 1.100: 6-Amino-2-butoxy-9-((6-(piperazin-1-yl)pyridin-3-yl)methyl)-9H-purin-8-ol (Compound 167)

[1065]

[1066] Compound 31 (15 mg, 0.041 mmol, 1.0 equiv), tert-butyl piperazine-1-carboxylate (9.2 mg, 0.050 mmol, 1.2 equiv), KOtBu (7.0 mg, 0.062 mmol, 1.5 equiv) and Pd-PEPPSI were placed in a crimp-sealable microwave vial. TMA solution of -iPr in DMF (0.40 mL) was thoroughly degassed and sealed under N2, then heated to 85°C for 2 h. The reaction mixture was then diluted with CH3CN / H2O (1 mL) and reverse phase flash purification was accomplished as described in General Procedure 5 using a 12 g C18 column with a gradient elution from 10% to 60% CH3CN+0.1% TFA / H2O+0.1% TFA to afford the title compound 167 (0.010 g, 0.025 mmol, 60%) as a white solid. LC-MS: C 19 H 26 Calculated m / z for N8O2 = 398.2, found [M+H] + =399.3. 1 H NMR (300MHz, MeOD) δ8.31(d,J=2.3Hz,1H),7.79(dd,J=8.8,2.4Hz,1H),6.95(d,J=8.8Hz,1H),4.94(s,2H),4.36(t,J=6 .5Hz,2H),3.86-3.76(m,4H),3.33-3.27(m,4H),1.85-1.70(m,2H),1.52(dq,J=14.5,7.4Hz,2H),1.01(t,J=7.4Hz,3H).

[1067] Example 1.101: 9-((6-(3,9-diazaspiro[5.5]undec-3-yl)pyridin-3-yl)methyl)-6-amino-2-butoxy-9H-purin-8-ol (Compound 168)

[1068]

[1069] To a solution of tert-butyl 3,9-diazaspiro[5.5]undecane-3-carboxylate (280 mg, 1.1 mmol, 10 equiv) in DMF (0.50 mL) and DIPEA- (50 μL) was added compound 31 (0.040 g, 0.11 mmol, 1.0 equiv) and the resulting mixture was heated to 140° C. for 18 h. The reaction was then adjusted to pH 1 with 6M HCl and preparative HPLC purification was accomplished as described in General Procedure 5 with a gradient elution of 10% to 65% CH 3 CN + 0.1% TFA / H 2 O + 0.1% TFA to afford the title compound 168 (0.7 mg, 0.0009 mmol, 0.8%) as a white solid. LC-MS: C 24 H 34 Calculated m / z for N8O2 = 466.3, found [M+H] + =467.4. 1H NMR (300MHz, MeOD) δ8.09(s,1H),8.01(d,J=8.2Hz,1H),7.25(d,J=9.4Hz,1H),4.93(s,2H),4.32(t,J=6.5Hz,2H),4.12(q ,J=7.0Hz,4H),3.67(t,J=5.9Hz,6H),3.23(t,J=5.9Hz,6H),1.78-1.62(m,2H),1.58-1.35(m,2H),1.00(t,J=7.4Hz,3H).

[1070] Example 1.102: 9-(4-Bromo-2-methoxybenzyl)-2-butoxy-8-methoxy-9H-purin-6-amine

[1071]

[1072] To a solution of compound 24 (580 mg, 2.4 mmol, 1.0 equiv) and 4-bromo-1-(bromomethyl)-2-methoxybenzene (680 mg, 2.4 mmol, 1.0 equiv) in DMF (8 mL) was added CsCO (0.80 g, 2.4 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 accomplished using a 60 g C18 column with a gradient elution from 10% to 60% CHCN + 0.1% TFA / H2O + 0.1% TFA as described in General Procedure 5. The title compound 32 (0.50 g, 1.1 mmol, 47%) was obtained as a white solid. LC-MS: C 18 H 22 Calculated m / z for BrN5O3 = 435.1, found [M+H] + =436.2.

[1073] Example 1.103: 6-Amino-2-butoxy-9-(2-methoxy-4-(piperazin-1-yl)benzyl)-9H-purin-8-ol (Compound 169)

[1074]

[1075] A solution of compound 32 (0.020 g, 0.046 mmol, 1.0 equiv), tert-butyl piperazine-1-carboxylate (11 mg, 0.060 mmol, 1.3 equiv), KOtBu (18 mg, 0.16 mmol, 3.5 equiv), Pd2dba3 (4.2 mg, 0.0046 mmol, 0.10 equiv), and RuPhos (11 mg, 0.023 mmol, 0.50 equiv) in DMF (0.50 mL) in a crimp-sealed microwave vial was thoroughly degassed and sealed under N2, then heated to 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 accomplished as described in General Procedure 5 with a gradient elution from 10% to 60% CH3CN + 0.1% TFA / H2O + 0.1% TFA to afford the Boc / 8-MeO intermediate. This intermediate was then dissolved in CH3CN (500 μL), H2O (500 μL) and 6M HCl (100 μL), and the mixture was heated to 55°C for 3 h. The reaction mixture was cooled to room temperature and lyophilized to afford the title product 169 (0.010 g, 0.015 mmol, 33%). LC-MS: C 21 H 29 Calculated m / z for N7O3 = 427.2, found [M+H] + =428.3. 1 H NMR (300MHz, MeOD) δ6.98(d,J=8.3Hz,1H),6.64(d,J=2.1Hz,1H),6.57-6.44(m,1H),4.94(s,2H),4.30(t,J= 6.5Hz, 2H), 3.88 (s, 3H), 3.39 (q, J = 6.1Hz, 8H), 1.80-1.69 (m, 2H), 1.56-1.36 (m, 2H), 0.96 (t, J = 7.4Hz, 3H).

[1076] Example 1.104: 6-Amino-9-(4-(4-aminopiperidin-1-yl)-2-methoxybenzyl)-2-butoxy-9H-purin-8-ol (Compound 170)

[1077]

[1078] A solution of compound 32 (15 mg, 0.034 mmol, 1.0 equiv), 4-Boc-aminopiperidine (10 mg, 0.052 mmol, 1.5 equiv), KOtBu (15 mg, 0.14 mmol, 4.0 equiv) and Pd2dba3 (6.3 mg, 0.0068 mmol, 0.20 equiv) and RuPhos (12 mg, 0.028 mmol, 0.80 equiv) in DMF (0.50 mL) in a crimp-sealed microwave vial was thoroughly degassed and sealed under N2, then heated to 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 accomplished as described in General Procedure 5 with a gradient elution from 10% to 60% CH3CN + 0.1% TFA / H2O + 0.1% TFA to provide the Boc / 8-MeO intermediate. This intermediate was then dissolved in CH3CN (500 μL) and 6M HCl (250 μL), and the mixture was heated to 55°C for 3 h. The reaction mixture was cooled to room temperature and lyophilized to give the title product 170 (1.8 mg, 0.0027 mmol, 7.9%). LC-MS: C 22 H 31 Calculated m / z for N7O3 = 441.3, found [M+H] + =442.4. 1 H NMR(300MHz,MeOD)δ6.96(d,J=8.3Hz,1H),6.62(s,1H),6.52(dd,J=8.4,2.2Hz,1H),4.96(s,2H),4.31(t,J=6.5Hz,2H),3.84(s ,3H),3.90-3.75(m,3H),2.86(t,J=12.4Hz,2H),2.12-2.07(m,2H),1.84-1.65(m,4H),1.56-1.38(m,2H),0.97(t,J=7.4Hz,3H).

[1079] Example 1.105: 6-Amino-2-butoxy-9-(2-methoxy-4-(3,9-diazaspiro[5.5]undec-3-yl)benzyl)-9H-purin-8-ol (Compound 171)

[1080]

[1081] 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-sealed microwave vial was thoroughly degassed and sealed under N2, then heated to 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 accomplished as described in General Procedure 5 with a gradient elution from 10% to 60% CH3CN + 0.1% TFA / H2O + 0.1% TFA to afford the Boc / 8-MeO intermediate. This intermediate was then dissolved in CH3CN (500 μL) and 6M HCl (250 μL), and the mixture was heated to 55°C for 3 h. The reaction mixture was cooled to room temperature and lyophilized to afford the title product 171 (6.4 mg, 0.0088 mmol, 29%). LC-MS: C 26 H 37 Calculated m / z for N7O3 = 495.3, found [M+H] + =496.4. 1 H NMR (300MHz, MeOD) δ7.20-7.10(m,2H),7.02(dd,J=8.2,2.2Hz,1H),5.03(s,2H),4.31(t,J=6.5Hz,2H),3.94(s,3H),3. 56(t,J=5.7Hz,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=7.4Hz,3H).

[1082] Example 1.106: 3-(4-((6-Amino-2-butoxy-8-methoxy-9H-purin-9-yl)methyl)-3-methoxy-phenyl)prop-2-yn-1-ol

[1083]

[1084] 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 with a rubber septum and thoroughly degassed with N2. Anhydrous propargyl alcohol (75 μL, 1.3 mmol, 1.5 equiv) was added via syringe and the reaction mixture was heated to 75°C under N2 for 18 h, after which it was diluted with H2O (50 mL) and extracted with EtOAc (3 x 50 mL). The pooled organics were then washed with brine (1 x 50 mL), dried over Na2SO4 and concentrated in vacuo to give the crude product. Reverse phase flash purification was accomplished as described in General Procedure 5 using a 60 g C18 column eluting with a gradient of 10% to 75% CH3CN + 0.1% TFA / H2O + 0.1% TFA to afford the title product 33 (130 mg, 0.32 mmol, 36%) as a dark red solid. LC-MS: C 21 H 25 Calculated m / z for N5O4 = 411.2, found [M+H] + =412.3.

[1085] Example 1.107: 3-(4-((6-Amino-2-butoxy-8-methoxy-9H-purin-9-yl)methyl)-3-methoxy-phenyl)propan-1-ol

[1086]

[1087] A solution of compound 33 (150 mg, 0.37 mmol, 1.0 eq) in MeOH (5 mL) was degassed with N2, after which 10% Pd / C (39 mg, 0.037 mmol, 0.10 eq) was added and the reaction vessel sealed and purged with additional N2. The reaction vessel was then purged with H2 and the mixture was stirred at room temperature under H2 for 18 h. The suspension was filtered through celite and the filtrate was concentrated in vacuo to give the title product 34 (130 mg, 0.30 mmol, 83%; continued) as a red oily solid that was carried forward without additional purification. LC-MS: C 21 H 29 Calculated m / z for N5O4 = 415.2, found [M+H] + =416.3.

[1088] Example 1.108: 3-(4-((6-amino-2-butoxy-8-hydroxy-9H-purin-9-yl)methyl)-3-methoxyphenyl)-propyl methanesulfonate

[1089]

[1090] To a solution of compound 34 (0.050 g, 0.12 mmol, 1.0 equiv) dissolved in anhydrous DCM (5 mL) was added DIPEA (63 μL, 0.36 mmol, 3.0 equiv) followed by methanesulfonyl chloride (17 mg, 0.14 mmol, 1.2 equiv) and the resulting solution was stirred at room temperature for 1 h. The solvent was removed in vacuo to afford the title product 35 (assuming quantitative yield) which was used further without additional purification. LC-MS: C 21 H 29 Calculated m / z for N5O6S = 479.2, found [M+H] + =480.3.

[1091] Example 1.109: 6-Amino-2-butoxy-9-(2-methoxy-4-(3-(piperazin-1-yl)propyl)benzyl)-9H-purin-8-ol (Compound 173)

[1092]

[1093] The title compound was prepared from compound 35 (0.010 g, 0.020 mmol, 1.0 equiv) and tert-butyl piperazine-1-carboxylate (15 mg, 0.081 mmol, 4.0 equiv) according to General Procedure 8. Preparative HPLC purification was accomplished as described in General Procedure 5 with a gradient elution from 10% to 60% CH3CN + 0.1% TFA / H2O + 0.1% TFA to afford the Boc / 8-MeO intermediate. This intermediate was then dissolved in CH3CN (400 μL) and 6M HCl (200 μL), and the mixture was heated to 55°C for 3 h. The reaction mixture was cooled to room temperature and lyophilized to afford the title product 173 (4.6 mg, 0.0057 mmol, 28%). LC-MS: C 24 H 35 Calculated m / z for N7O3 = 469.3, found [M+H] + =470.4. 1H NMR(300MHz,MeOD)δ7.02-6.85(m,2H),6.83-6.71(m,1H),5.01(s,2H),4.30(t,J=6.5Hz,2H),4.16(s,2H),3.86(s,3H),3.56-3.4 3(m,4H),3.06-2.93(m,2H),2.71(t,J=7.4Hz,2H),2.10-1.94(m,2H),1.77-1.64(m,2H),1.63-1.28(m,4H),0.97(t,J=7.4Hz,3H).

[1094] Example 1.110: 6-Amino-9-(4-(3-(4-aminopiperidin-1-yl)propyl)-2-methoxybenzyl)-2-butoxy-9H-purin-8-ol (Compound 172)

[1095]

[1096] The title compound was prepared from compound 35 (5.1 mg, 0.010 mmol, 1.0 equiv) and 4-Boc-aminopiperidine (8.3 mg, 0.042 mmol, 4.0 equiv) according to General Procedure 8. Preparative HPLC purification was accomplished as described in General Procedure 5 with a gradient elution from 10% to 60% CH3CN + 0.1% TFA / H2O + 0.1% TFA to afford the Boc / 8-MeO intermediate. This intermediate was then dissolved in CH3CN (400 μL) and 6M HCl (200 μL), and the mixture was heated to 55°C for 3 h. The reaction mixture was cooled to room temperature and lyophilized to afford the title product 172 (1.0 mg, 0.0012 mmol, 12%). LC-MS: C 25 H 37 Calculated m / z for N7O3 = 483.3, found [M+H] + =484.4. 1 H NMR(300MHz,MeOD)δ6.96-6.85(m,2H),6.76(d,J=7.7Hz,1H),5.00(s,2H),4 .27(t,J=6.5Hz,2H),3.88(s,3H),3.75-3.65(m,2H),3.47-3.41(m,1H),3.1 9-3.01(m,2H),2.77-2.66(m,2H),2.33-2.22(m,2H),2.11-2.05(m,2H),1.9 8-1.88(m,2H),1.79-1.63(m,2H),1.48-1.34(m,4H),0.97(t,J=7.4Hz,3H).

[1097] Example 1.111: 6-Amino-9-(4-(3-(4-aminopiperidin-1-yl)propyl)-2-methoxybenzyl)-2-butoxy-9H-purin-8-ol (Compound 174)

[1098]

[1099] The title compound was prepared from compound 35 (0.010 g, 0.020 mmol, 1.0 equiv) and tert-butyl 3,9-diazaspiro[5.5]undecane-3-carboxylate (0.020 g, 0.081 mmol, 4.0 equiv) according to General Procedure 8. Preparative HPLC purification was accomplished as described in General Procedure 5 with a gradient elution from 10% to 60% CH3CN+0.1%TFA / H2O+0.1%TFA to afford the Boc / 8-MeO intermediate. This intermediate was then dissolved in CH3CN (400 μL) and 6M HCl (200 μL), and the mixture was heated to 55°C for 3 h. The reaction mixture was cooled to room temperature and lyophilized to afford the title product 174 (4.6 mg, 0.0052 mmol, 26%). LC-MS: C 29 H 43 Calculated m / z for N7O3 = 537.3, found [M+H] + =538.4. 1 H NMR (300MHz, MeOD) δ6.95-6.85(m,2H),6.76(d,J=7.8Hz,1H),5.00(s,2H),4.27(t,J=6.6Hz,2H),3.88(s,3H),3.51-3.41(m,2H),3.28-3.18(m ,4H),3.15-3.09(m,2H),2.77-2.66(m,2H),2.13-1.93(m,6H),1.93-1. 87(m,4H),1.72-1.66(m,4H),1.56-1.37(m,2H),0.95(q,J=9.9Hz,3H).

[1100] 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)

[1101]

[1102] The title compound was prepared from compound 35 (0.010 g, 0.020 mmol, 1.0 equiv) and tert-butyl N-[2-(piperidin-4-yl)ethyl]carbamate (19 mg, 0.081 mmol, 4.0 equiv) according to General Procedure 8. Preparative HPLC purification was accomplished as described in General Procedure 5 with a gradient elution from 10% to 60% CH3CN + 0.1% TFA / H2O + 0.1% TFA to afford the Boc / 8-MeO intermediate. This intermediate was then dissolved in CH3CN (400 μL) and 6M HCl (200 μL), and the mixture was heated to 55°C for 3 h. The reaction mixture was cooled to room temperature and lyophilized to afford the title product 175 (1.6 mg, 0.0019 mmol, 9.4%). LC-MS: C 27 H 41 Calculated m / z for N7O3 = 511.3, found [M+H] + =512.4. 1 H NMR(300MHz,MeOD)δ6.94-6.85(m,2H),6.76(d,J=7.7Hz,1H),5.00(s,2H),4.25(t,J=6.6Hz,2H),3.88(s,3H),3.64-3.58(m,2H),3.2 1-2.85(m,6H),2.74-2.66(m,2H),2.21-1.93(m,4H),1.80-1.57(m,5H),1.53-1.39(m,2H),1.36-1.28(m,2H),0.96(t,J=7.4Hz,3H).

[1103] Example 1.113: 6-Amino-2-butoxy-9-(2-methoxy-4-(3-(piperidin-4-ylamino)propyl)benzyl)-9H-purin-8-ol (Compound 176)

[1104]

[1105] The title compound was prepared from compound 35 (0.010 g, 0.020 mmol, 1.0 equiv) and 1-Boc-aminopiperidinyl carbamate (19 mg, 0.081 mmol, 4.0 equiv) according to General Procedure 8. Preparative HPLC purification was accomplished as described in General Procedure 5 with a gradient elution from 10% to 60% CH3CN + 0.1% TFA / H2O + 0.1% TFA to afford the Boc / 8-MeO intermediate. This intermediate was then dissolved in CH3CN (400 μL) and 6M HCl (200 μL), and the mixture was heated to 55°C for 3 h. The reaction mixture was cooled to room temperature and lyophilized to afford the title product 176 (1.5 mg, 0.0018 mmol, 9.1%). LC-MS: C 25 H 37 Calculated m / z for N7O3 = 483.3, found [M+H] + =484.4. 1 H NMR (300MHz, MeOD) δ7.02-6.84(m,2H),6.76(dd,J=7.8,1.5Hz,1H),5.01(s,2H),4.30(t,J=6.5Hz,2H),3.87(s,3H),3.56-3.41(m,4H),3. 31-3.25(m,5H),3.06-2.92(m,2H),2.71(t,J=7.4Hz,2H),2.09-1.93(m,2H),1.84-1.64(m,2H),1.56-1.28(m,2H),0.97(t,J=7.4Hz,3H).

[1106] Example 1.114: Ethyl 2-(3-methoxy-4-methylphenyl)acetate

[1107]

[1108] To a solution of (3-methoxy-4-methylphenyl)acetic acid (5.0 g, 28 mmol, 1.0 equiv) in EtOH (30 mL) was added concentrated H2SO4 (0.3 mL). The resulting mixture was heated to reflux and maintained for 1.5 h, then concentrated in vacuo and redissolved in EtOAc (50 mL). The resulting organic solution was extracted with H2O (1 x 20 mL), then with brine (1 x 10 mL), dried over MgSO4 and concentrated in vacuo to give the crude title product 36 as a yellow oil that was carried forward without additional purification (assuming quantitative yield). LC-MS: C 12 H 16 Calculated m / z for O3 = 208.1, found [M+H] + =209.2.

[1109] Example 1.115: Ethyl 2-(4-(bromomethyl)-3-methoxyphenyl)acetate

[1110]

[1111] To a solution of compound 36 (2.7 g, 13 mmol, 1.0 equiv) in DCM (10 mL) was added NBS (2.5 g, 14 mmol, 1.0 equiv) followed by 30% H2O2 (1.5 g, 13 mmol, 1.0 equiv). The resulting solution was stirred at room temperature for 18 h, concentrated in vacuo and redissolved in CH3CN (5 mL) and H2O (1 mL). Reverse phase flash purification was performed as described in General Procedure 5 using a 60 g C18 column eluting with a gradient of 20% to 60% CH3CN+0.1% TFA / H2O+0.1% TFA, followed by normal phase flash purification using a 50 g silica column eluting with a gradient of 10% to 100% EtOAc / hexanes as described in General Procedure 5 to afford the title compound 37 (350 mg, 1.2 mmol, 9.4%). LC-MS: C 12 H 15 Calculated m / z for BrO3 = 286.0, found [M+H] + =287.0.

[1112] Example 1.116: Ethyl 2-(4-((6-amino-2-butoxy-8-methoxy-9H-purin-9-yl)methyl)-3-methoxy-phenyl)acetate

[1113]

[1114] To a solution of compound 24 (470 mg, 2.0 mmol, 1.0 equiv) and compound 37 (560 mg, 2.0 mmol, 1.0 equiv) in DMF (3 mL) was added CsCO (640 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 rapid purification was accomplished using a 60 g C18 column with a gradient elution from 10% to 50% CHCN + 0.1% TFA / H2O + 0.1% TFA as described in General Procedure 5. The title compound 38 (160 mg, 0.35 mmol, 18%) was obtained as a white solid. LC-MS: C 22 H 29 Calculated m / z for N5O5 = 443.2, found [M+H] + =444.3.

[1115] Example 1.117: 2-(4-((6-Amino-2-butoxy-8-methoxy-9H-purin-9-yl)methyl)-3-methoxy-phenyl)ethan-1-ol

[1116]

[1117] To a stirred solution of compound 38 (380 mg, 0.68 mmol, 1.0 equiv) in anhydrous THF (2 mL) cooled to 0°C was added lithium aluminum hydride (86 mg, 1.4 mmol, 2.0 equiv). The resulting suspension was allowed to warm to room temperature and stirred for 15 min before being diluted with MeOH (5 mL) and H2O (1 mL) and filtered through a plug of celite. The filtrate was then concentrated in vacuo and reverse phase flash purification was accomplished as described in General Procedure 5 using a 30 g C18 column eluting with a gradient of 10% to 50% CH3CN+0.1% TFA / H2O+0.1% TFA to afford the title product 39 (0.20 g, 0.39 mmol, 57%) as a white solid. LC-MS: C 20 H 27 Calculated m / z for N5O4 = 401.2, found [M+H] + =402.3.

[1118] Example 1.118: 6-Amino-2-butoxy-9-(4-(2-chloroethyl)-2-methoxybenzyl)-9H-purin-8-ol

[1119]

[1120] The title compound 40 was prepared from compound 39 (0.10 g, 0.19 mmol, 1.0 eq) and 10% SOCl2 / DCM (10 mL) according to General Procedure 3. The title compound was obtained as a yellow solid (72 mg, 0.16 mmol, 84%). LC-MS: C 19 H 24 Calculated m / z for ClN5O3 = 405.2, found [M+H] + =406.3.

[1121] Example 1.119: 6-Amino-2-butoxy-9-(2-methoxy-4-(2-(piperazin-1-yl)ethyl)benzyl)-9H-purin-8-ol (Compound 180)

[1122]

[1123] The title compound was prepared according to General Procedure 1 from compound 40 (9.7 mg, 0.024 mmol, 1.0 equiv), NaI (0.5 mg, 0.003 mmol, 0.1 equiv) and tert-butyl piperazine-1-carboxylate (9.0 mg, 0.048 mmol, 2.0 equiv). Preparative HPLC purification was accomplished as described in General Procedure 5 with a gradient elution from 10% to 60% CH3CN + 0.1% TFA / H2O + 0.1% TFA. Deprotection was accomplished according to General Procedure 2 to afford the title compound 180 (7.0 mg, 0.010 mmol, 43%) as a white solid. LC-MS: C 23 H 33 Calculated m / z for N7O3 = 455.3, found [M+H] + =456.4. 1 H NMR (300MHz, MeOD) δ7.23(dd,J=8.5,2.2Hz,1H),7.03-6.97(m,2H),5.04(s,2H),4.35(td,J=6.5,4.1Hz,2H),3.86(d,J=2.0Hz,3H),3. 58-3.43(m,6H),3.32-3.19(m,4H),2.96(dd,J=10.0,6.1Hz,2H),1.74(dq,J=8.5,6.6Hz,2H),1.56-1.38(m,2H),0.97(t,J=7.4Hz,3H).

[1124] Example 1.120: 6-Amino-9-(4-(2-(4-aminopiperidin-1-yl)ethyl)-2-methoxybenzyl)-2-butoxy-9H-purin-8-ol (Compound 181)

[1125]

[1126] The title compound was prepared from compound 40 (9.7 mg, 0.024 mmol, 1.0 equiv), NaI (0.5 mg, 0.003 mmol, 0.1 equiv) and 4-Boc-aminopiperidine (0.010 g, 0.048 mmol, 2.0 equiv) according to General Procedure 1. Preparative HPLC purification was accomplished as described in General Procedure 5 with a gradient elution from 10% to 60% CH3CN + 0.1% TFA / H2O + 0.1% TFA. Deprotection was accomplished according to General Procedure 2 to afford the title compound 181 (6.1 mg, 0.0087 mmol, 37%) as a white solid. LC-MS: C 24 H 35 Calculated m / z for N7O3 = 469.3, found [M+H] + =470.4.1 H NMR (300MHz, MeOD) δ7.23(dd,J=8.3,2.3Hz,1H),7.04-6.93(m,2H),5.04(s,2H),4.35(t,J=2.2Hz,2H),3.85(s,3H),3.79-3.39(m,3 H),3.33-3.05(m,4H),2.98(dd,J=10.3,6.4Hz,2H),2.38-1.89(m,4H),1.82-1.58(m,2H),1.56-1.38(m,2H),0.97(t,J=7.4Hz,3H).

[1127] Example 1.121: 9-(4-(2-(3,9-diazaspiro[5.5]undec-3-yl)ethyl)-2-methoxybenzyl)-6-amino-2-butoxy-9H-purin-8-ol (Compound 177)

[1128]

[1129] The title compound was prepared according to General Procedure 1 from compound 40 (9.7 mg, 0.024 mmol, 1.0 equiv), NaI (4 mg, 0.02 mmol, 1 equiv) and tert-butyl 3,9-diazaspiro[5.5]undecane-3-carboxylate (18 mg, 0.071 mmol, 3.0 equiv). Preparative HPLC purification was accomplished as described in General Procedure 5 with a gradient elution from 10% to 60% CH 3 CN + 0.1% TFA / H 2 O + 0.1% TFA. Deprotection was accomplished according to General Procedure 2 to afford the title compound 177 (11 mg, 0.015 mmol, 63%) as a white solid. LC-MS: C 28 H 41 Calculated m / z for N7O3 = 523.3, found [M+H] + =524.4. 1 H NMR(300MHz,MeOD)δ7.23(dd,J=8.4,2.2Hz,1H),7.04-6.96(m,2H),5.04(s,2H),4.37(t,J=6.5Hz,2H),3.86(s,3H),3.55-3.4 5(m,2H),3.34-3.05(m,8H),3.03-2.91(m,2H),2.08-1.86(m,4H),1.83-1.64(m,6H),1.60-1.29(m,2H),0.97(t,J=7.4Hz,3H).

[1130] 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)

[1131]

[1132] The title compound was prepared according to General Procedure 1 from compound 40 (9.7 mg, 0.024 mmol, 1.0 equiv), NaI (4 mg, 0.02 mmol, 1 equiv) and tert-butyl N-[2-(piperidin-4-yl)ethyl]carbamate (16 mg, 0.071 mmol, 3.0 equiv). Preparative HPLC purification was accomplished as described in General Procedure 5 with a gradient elution from 10% to 60% CH3CN+0.1% TFA / H2O+0.1% TFA. Deprotection was accomplished according to General Procedure 2 to afford the title compound 178 (13 mg, 0.018 mmol, 76%) as a white solid. LC-MS: C 26 H 39 Calculated m / z for N7O3 = 497.3, found [M+H] + =498.4. 1 H NMR(300MHz,MeOD)δ7.26-7.10(m,1H),7.04-6.88(m,1H),6.94(s,1H),5.03(t,J=2 .4Hz,2H),4.40-4.24(m,2H),3.83(s,3H),3.72-3.58(m,2H),3.26(dd,J=10.4,6.3 Hz,2H),2.97(dt,J=18.0,7.2Hz,4H),2.71(t,J=7.1Hz,2H),2.02(d,J=14.1Hz,1H) ,1.81-1.59(m,6H),1.45(p,J=7.5Hz,2H),1.34-1.28(m,2H),0.97(t,J=7.4Hz,3H).

[1133] Example 1.123: 6-Amino-2-butoxy-9-(2-methoxy-4-(2-(piperidin-4-ylamino)ethyl)benzyl)-9H-purin-8-ol (Compound 179)

[1134]

[1135] The title compound was prepared according to General Procedure 1 from compound 40 (9.7 mg, 0.024 mmol, 1.0 equiv), NaI (4 mg, 0.02 mmol, 1 equiv) and 1-Boc-aminopiperidine (0.010 g, 0.048 mmol, 2.0 equiv). Preparative HPLC purification was accomplished as described in General Procedure 5 with a gradient elution from 10% to 60% CH 3 CN + 0.1% TFA / H 2 O + 0.1% TFA. Deprotection was accomplished according to General Procedure 2 to afford the title compound 179 (12 mg, 0.015 mmol, 62%) as a white solid. LC-MS: C 24 H 35 Calculate...

Claims

1. A compound of formula (I): Its tautomers, protected forms and / or pharmaceutically acceptable salts, wherein: X is O or NH; Y is N or CH; R 1 is an optionally substituted C2-C6 alkyl group or a branched and optionally substituted C3-C8 hydroxyalkyl group; R 2 is H, halogen, optionally substituted C1-C6 alkyl, or optionally substituted C1-C6 alkoxy; R 3 and R 4 Independently for H, QR 5 , or R 3 and R 4 Together with N, it forms an optionally substituted C2-C6-heterocycloalkyl ring, an optionally substituted C3-C7-heterobicycloalkyl ring, an optionally substituted C6-C 10 - a heterospirocycloalkyl ring or an optionally substituted heteroaryl ring; Q is a bond, optionally substituted C1-C6-alkyl, optionally substituted C2-C6-alkenyl or Where * is the same as R 5 , and # is the attachment point with N; R 5 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, it forms an optionally substituted C2-C6-heterocycloalkyl ring, an optionally substituted C3-C7-heterobicycloalkyl ring, an optionally substituted C6-C 10 - a 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, optionally substituted aryl or optionally substituted heteroaryl; n is an integer from 1 to 4; m is an integer from 0 to 4; and a and b are each independently an integer from 0 to 4, The condition is that when R 2 When it is H, then: (i)R 1 is a branched and optionally substituted C3-C8 hydroxyalkyl group; (ii) X is NH and R 1 is an optionally substituted C5-C6 alkyl group; or (iii) m is 0 or 1, and R 3 and R 4 Together with N, it forms an unsubstituted piperazinyl ring or an optionally substituted C6-C 10 -heterospirocycloalkyl ring. The compound according to claim 1, wherein R 2 is halogen, optionally substituted C1-C6 alkyl, or optionally substituted C1-C6 alkoxy.

2. The compound of claim 1, wherein R 2 is halogen, optionally substituted C1-C6 alkyl, or optionally substituted C1-C6 alkoxy.

3. A compound as described in any one of claims 1 to 2, wherein R 2 is halogen or optionally substituted C1-C6 alkoxy.

4. The compound of any one of claims 1 to 3, wherein R 2 is halogen or unsubstituted C1-C6 alkoxy.

5. The compound of any one of claims 1 to 4, wherein R 2 is fluorine or methoxy.

6. The compound of any one of claims 1 to 5, wherein X is O.

7. The compound of any one of claims 1 to 5, wherein X is NH.

8. A compound as described in any one of claims 1 to 7, wherein R 1 is an optionally substituted C2-C6 alkyl group.

9. The compound of any one of claims 1 to 7, wherein R 1 is a branched and optionally substituted C3-C8 hydroxyalkyl group.

10. The compound of any one of claims 1 to 9, wherein R 3 and R 4 H or QR independently 5 .

11. The compound of any one of claims 1-10, wherein n is 1.

12. The compound of any one of claims 1-11, wherein m is an integer from 0 to 3 or from 1 to 3.

13. The compound of claim 1, wherein R 2 H and R 1 is a branched and optionally substituted C3-C8 hydroxyalkyl group.

14. The compound of claim 1, wherein the compound is selected from any one of compounds 100-195 listed in Table 1.

15. The compound according to claim 1, which has the structure of formula (II): in: X is O or NH; Y is N or CH; R 1 is an optionally substituted C2-C6 alkyl group or a branched and optionally substituted C3-C8 hydroxyalkyl group; R 3 and R 4 Independently for H, QR 5 , or R 3 and R 4 Together with N, it forms an optionally substituted C2-C6-heterocycloalkyl ring, an optionally substituted C3-C7-heterobicycloalkyl ring, an optionally substituted C6-C 10 - a heterospirocycloalkyl ring or an optionally substituted heteroaryl ring; Q is a bond, optionally substituted C1-C6-alkyl, optionally substituted C2-C6-alkenyl or Where * is the same as R 5 , and # is the attachment point with N; R 5 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, it forms an optionally substituted C2-C6-heterocycloalkyl ring, an optionally substituted C3-C7-heterobicycloalkyl ring, an optionally substituted C6-C 10 - a 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, optionally substituted aryl or optionally substituted heteroaryl; m is an integer from 0 to 4; and a and b are independently integers from 0 to 4.

16. The compound of claim 15, wherein the compound is selected from Subtable 1A.

17. The compound of any one of claims 1-9, wherein the compound is selected from Subtable 1B.

18. The compound of claim 15, wherein the compound is:

19. The compound of claims 1-10, wherein R 3 and R 4 Both are H.

20. The compound of claim 19, wherein the compound is:

21. The compound of claim 1, wherein the compound is selected from Subtable 1C.

22. The compound of claim 1 or claim 15, wherein the compound is:

23. The compound of any one of claims 1-10, wherein the compound is selected from Subtable ID.

24. A compound as described in any one of claims 1 to 10, wherein R 6 H and R 7 is H, an optionally substituted C1-C6 alkyl group, or R 6 and R 7 Together with N, it 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.

25. The compound of claim 24, wherein the compound is selected from Subtable 1E.

26. The compound of any one of claims 1 to 9, wherein the compound is:

27. The compound of claim 1 or claim 15, wherein the compound is:

28. A compound of formula (III): Its tautomers, protected forms and / or pharmaceutically acceptable salts, in: R 1a is an optionally substituted C2-C6 alkyl group or a branched and optionally substituted C3-C8 hydroxyalkyl group; R 2a is halogen, optionally substituted C1-C6 alkyl, or optionally substituted C1-C6 alkoxy; R 3a and R 4a Independently for H, QR 5a , or R 3a and R 4a Together with N, it forms an optionally substituted C2-C6-heterocycloalkyl ring, an optionally substituted C3-C7-heterobicycloalkyl ring, an optionally substituted C6-C 10 - a heterospirocycloalkyl ring or an optionally substituted heteroaryl ring; Q is a bond, optionally substituted C1-C6-alkyl, optionally substituted C2-C6-alkenyl or Where * is the same as R 5 , and # is the attachment point with N; R 5a 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; R 6a and R 7a are independently H, optionally substituted C1-C6 alkyl, optionally substituted C1-C6 alkoxycarbonyl, or R 6a and R 7a Together with N, it forms an optionally substituted C2-C6-heterocycloalkyl ring, an optionally substituted C3-C7-heterobicycloalkyl ring, an optionally substituted C6-C 10 - a heterospirocycloalkyl ring or an optionally substituted heteroaryl ring; W is an optionally substituted C3-C6-cycloalkyl, an optionally substituted C3-C6-heterocycloalkyl, an optionally substituted C5-C7-heterobicycloalkyl, an optionally substituted C6-C 10 -heterospirocycloalkyl, optionally substituted aryl or optionally substituted heteroaryl; u is an integer from 1 to 3; v is an integer from 0 to 4; and a and b are independently integers from 0 to 4.

29. The compound of claim 28, having the structure of formula (IV): in: R 1a is an unsubstituted C2-C6 alkyl group or a branched and unsubstituted C3-C8 hydroxyalkyl group; R 3a and R 4a Independently for H, QR 5a , or R 3a and R 4a Together with N, it forms an optionally substituted C2-C6-heterocycloalkyl ring, an optionally substituted C3-C7-heterobicycloalkyl ring, an optionally substituted C6-C 10 - a heterospirocycloalkyl ring or an optionally substituted heteroaryl ring; Q is a bond, optionally substituted C1-C6-alkyl, optionally substituted C2-C6-alkenyl or Where * is the same as R 5 , and # is the attachment point with N; R 5a 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; R 6a and R 7a are independently H, optionally substituted C1-C6 alkyl, optionally substituted C1-C6 alkoxycarbonyl, or R 6a and R 7a Together with N, it forms an optionally substituted C2-C6-heterocycloalkyl ring, an optionally substituted C3-C7-heterobicycloalkyl ring, an optionally substituted C6-C 10 - a 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, optionally substituted aryl or optionally substituted heteroaryl; v is an integer from 0 to 4; and a and b are independently integers from 0 to 4.

30. compounds as claimed in claim 28 or claim 29, wherein each alkyl, alkenyl, cycloalkyl, spiroalkyl, heterocycloalkyl, heterospiroalkyl, aryl and heteroaryl are optionally replaced by -NH , -CO H , -OH, carbonyl, halogen, C -C alkyl, C -C hydroxyalkyl, C -C aminoalkyl, C -C carboxylalkyl, (C -C alkyl)-cycloalkyl, (C -C alkyl)-heterocycloalkyl, (C -C alkyl)-spiroalkyl, (C -C alkyl)-heterospiroalkyl, (C -C alkyl)- -C4 alkyl)-aryl and (C0-C4 alkyl)-heteroaryl, and wherein each of the alkyl, hydroxyalkyl, C1-C4 aminoalkyl, C1-C4 carboxyalkyl, cycloalkyl, heterocycloalkyl, spirocycloalkyl, heterospirocycloalkyl, aryl or heteroaryl can itself be substituted by one or more of the following: -NH2, -CO2H, -OH, carbonyl, halogen, unsubstituted C1-C4 alkyl, unsubstituted C1-C4 hydroxyalkyl, unsubstituted C1-C4 aminoalkyl or unsubstituted C1-C4 carboxyalkyl.

31. compounds as claimed in claim 30, wherein each alkyl, alkenyl, cycloalkyl, spirocycloalkyl, heterocycloalkyl, heterospirocycloalkyl, aryl and heteroaryl are optionally substituted by 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.

32. The compound of claim 28 or claim 29, wherein the compound is selected from Sub-Table 1A.

33. A compound as claimed in claim 28 or claim 29, wherein the compound is:

34. The compound of any one of claims 1-33, wherein the compound has an EC for agonizing human or mouse TLR7 of <500 nM, <250 nM, or <100 nM as determined in a reporter gene assay. 50 Values, and / or EC values ​​of <5 μM, <1 μM, <500 nM or <100 nM for inducing IL6 production in a human or murine immune cell population as determined in an immune cell assay 50 value.

35. A pharmaceutical composition comprising a compound according to any one of claims 1 to 34 and a pharmaceutically acceptable carrier or diluent.

36. A compound as claimed in any one of claims 1 to 34 for use in therapy.

37. A compound as claimed in any one of claims 1 to 34 for use in the treatment of cancer.

38. Use of a compound as claimed in any one of claims 1 to 34 in the manufacture of a medicament for the treatment of cancer.

39. A compound-linker construct having formula (A): (A) L-(C) p in: L is the connector; C is a compound according to any one of claims 1 to 34; and p is an integer of 1-5.

40. The compound-linker construct of claim 39, wherein L is a cleavable linker.

41. The compound-linker construct of claim 39, wherein L is a non-cleavable linker.

42. The compound-linker construct of any one of claims 39-40, wherein L comprises a dipeptide, a tripeptide, a tetrapeptide, or a combination thereof.

43. The compound-linker construct of any one of claims 39-42, wherein L comprises a polyethylene glycol (PEG) moiety.

44. The compound-linker construct of any one of claims 39-40, wherein L is a protease cleavable linker.

45. The compound-linker construct of any one of claims 39-44, wherein C is the compound of claim 36.

46. ​​The compound-linker construct of any one of claims 39-44, wherein C is the compound of claim 66.

47. The compound-linker construct of claim 39, wherein the compound-linker is selected from Table 2B.

48. The compound-linker construct of any one of claims 39-46, wherein p is 1, 2 or 3.

49. A conjugate having formula (X): (X) T-[L-(C) p ] r in: T is the targeting moiety; L is the connector; C is a compound according to any one of claims 1 to 34; p is an integer from 1 to 5; and The value of r is from about 1 to about 8.

50. The conjugate of claim 49, wherein L-(C)p is the compound-linker construct of any one of claims 39-48.

51. The conjugate of any one of claims 49-50, wherein 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.

52. The conjugate of claim 49, wherein C is selected from Subtable 1F.

53. The conjugate of any one of claims 49-52, wherein T binds to a tumor associated antigen (TAA).

54. The conjugate of any one of claims 49-53, wherein T is an antibody or an antigen-binding antibody fragment.

55. The conjugate of claim 54, wherein the antibody is a bispecific or multispecific antibody.

56. The conjugate of any one of claims 49-55, wherein the conjugate has an EC for inducing cytokine production in a human or murine immune cell population of <1 nM, <500 pM or <100 pM as determined in a reporter gene assay. 50 value.

57. A pharmaceutical composition comprising the conjugate of any one of claims 49-56 and a pharmaceutically acceptable carrier or diluent.

58. The conjugate of any one of claims 49-56 for use in therapy.

59. The conjugate of any one of claims 49-56 for use in cancer treatment.

60. Use of the conjugate of any one of claims 49-56 in the manufacture of a medicament for the treatment of cancer.

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