Compounds, compositions, and methods for treating fibrotic diseases and cancer

Abstract

Compounds comprising Toll-like receptor (TLR) agonists, e.g., TLR7 and TLR7 / 8 agonists, and folic acid or pteroyl amino acid conjugates thereof, and their use for treating cancer or fibrotic diseases or disorders; and methods for preparing conjugates comprising folate receptor targeting ligands and TLR7 and TLR7 / 8 agonists.

Description

Technical Field

[0001] Priority This application claims the benefit of priority of U.S. Provisional Patent Application No. 63 / 049,556, filed Jul. 8, 2020, the entire contents of which are incorporated herein by reference.

[0002] The present disclosure relates to compounds, compositions, and methods for treating various diseases, including, for example, cancer, fibrosis, and other disease states. In some embodiments, the present disclosure relates to Toll-like receptor (TLR) agonists, such as TLR-7 / 8 agonists and folic acid or pteroyl amino acid conjugates thereof, and their use for treating cancer and inflammatory diseases (e.g., fibrotic diseases). In some embodiments, the present disclosure further relates generally to methods of preparing conjugates comprising a targeting ligand of the folate receptor and a TLR7 / 8 agonist.

Background Art

[0003] Macrophages are an important cellular component of innate immunity. M1 macrophages are pro-inflammatory, while M2 macrophages are anti-inflammatory cells. Overstimulation of M1-like and M2-like macrophages has been associated with several diseases, such as fibrosis, inflammatory diseases, and cancer. In cancer, tumor-associated macrophages (TAMs) with an M2 phenotype become the most prominent cells in the tumor microenvironment (TME). Increased TAM infiltration has been associated with poor prognosis in many cancers.In the TME, TAMs contribute to immunosuppressive functions. In fibrotic diseases, activated M2-like macrophages produce profibrotic cytokines that induce the production of extracellular matrix proteins, including collagen and fibronectin, by myofibroblasts. Effective treatment for such diseases may be provided by reprogramming these immunosuppressive phenotypes into more pro-inflammatory phenotypes.

[0004] Toll-like receptors (TLRs) can recognize pathogens and are significantly expressed in immune cells. Synthetic small molecule agonists that target TLR-7 / 8 are known to function as potent immunostimulants. However, systemic administration of such TLR-7 / 8 agonists in non-targeted forms may be hindered by dose-limiting toxicity and may cause toxic cytokine syndrome in humans. Therefore, many such drugs are administered topically. [Overview of the Initiative]

[0005] In certain embodiments, compounds and compositions are provided herein for use, for example, in therapeutic methods. In some examples, compounds and compositions are used in treatments, such as methods for treating cancer and / or fibrosis. In some embodiments, the compounds are Toll-like receptor (TLR) 7 and / or 8 agonists. In certain embodiments, the compounds are used alone or in combination with a targeting agent.

[0006] The compounds provided herein may include a first radical. The first radical may be linked to a second radical (e.g., directly or via a linker). The second radical may be a targeting moiety that targets a cellular pattern recognition receptor (e.g., an immune cell receptor, e.g., a folate receptor, e.g., folate receptor beta (FR-β)). In some embodiments, the targeting ligand includes a folate receptor-binding ligand, e.g., folate or a functional fragment or analog thereof, e.g., a pteroyl amino acid (e.g., a pteroyl linked to an amino acid or a peptide containing two or more amino acids).

[0007] In some cases, for example, if the compound is a (e.g., potent) TLR7 / 8 agonist, the unconjugated compound can be extremely toxic if delivered systemically. It is desirable to reduce and / or eliminate the systemic toxicity associated with such compounds. The conjugated radical of a compound may have reduced toxicity compared to the free form of the compound (e.g., at least 10%, at least 20%, at least 30%, at least 50%, at least 75%, or at least 90% reduction). Furthermore, in some cases, the compound (conjugate) may be effective at concentrations equivalent to or lower than the free form of the compound (e.g., having an ED50 concentration of ≤120%, ≤100%, ≤80%, ≤60%, or ≤40% of the free form).

[0008] In certain embodiments, the compound comprises a first radical connected to a second radical via a non-releasing bond, for example, via a non-releasing linker. The non-releasing bond of the compound or its analogues in the conjugate can reduce systemic exposure to the compound (e.g., corresponding toxicity).

[0009] In certain embodiments, the second (e.g., targeted) radical is folic acid or an analog, functional fragment, or derivative (e.g., a pteroyl amino acid). In some examples, such ligands are useful for targeting cellular pattern recognition receptors, such as FR-β. In some cases, FR-β is overexpressed in activated myeloid cells, but is present at very low levels in healthy cells.

[0010] In some embodiments, the first radical provided herein is a TLR7 and / or 8 (TLR7 / 8) agonist, e.g., a potent agonist. In some embodiments, delivery of a TLR-7 / 8 agonist via a targeted ligand (e.g., folic acid or pteroyl amino acid) has been demonstrated to be effective in mitigating systemic cytokine release.

[0011] In some embodiments, folic acid-conjugated TLR-7 / 8 agonists provide specificity for diseased cell types. In one embodiment, for example, a folic acid-TLR7 / 8 agonist conjugate can be delivered (e.g., specifically) into the endosomes of FR-β+ macrophages while limiting systemic exposure to the TLR7 / 8 agonist.

[0012] In some examples, direct alkylation of tertiary hydroxyls with alkyl halides generally proceeds in low yield and with positional isomer products due to the steric effects of the bulky tert-butyl group. In some embodiments, compounds containing radicals (e.g., TLR7 agonist radicals) couple to folic acid (radical) with higher efficiency compared to more sterically hindered compounds. In some embodiments, alkylene spacers (e.g., n>0, e.g. n=1-8, e.g. n=1, forming a methylene group) are positioned between the tert-butyl group and the hydroxyl group (e.g., illustrated in formula I below), enabling efficient chemical synthesis and resulting in stable conjugates in high yield.

[0013] [ka]

[0014] The advantages of such compounds are, in some examples, that such compounds (their radicals) form stable conjugates with folic acid ligands or functional fragments or analogs thereof connected via a non-releasing linker. In some embodiments, for example when Y is hydroxyl, the connection to the linker results in the formation of an ester (-OCO-), carbonate (-OC(=O)O-), or carbamate (-OC(=O)NR-) at Y. In other examples, Y is another group described herein. In some embodiments, the incorporation of the spacer enables the conjugate having a radical of formula (I) connected to a folic acid receptor ligand via a non-releasing linker to form a stable adduct. In some examples, the conjugate is more stable (e.g., in vivo), reducing the systemic exposure of the TLR7 agonist and, for example, reducing the adverse effect and side effect profile.

[0015] In some embodiments, a compound represented by the structure of formula (I) (or comprising a radical of the structure of formula (I)):

[0016]

Chemical formula

[0017]

Chemical formula

[0018] ]>

Chemical formula

[0019]

Chem.

[0021] In some embodiments, one or more compounds of formula I are R 3 Base (for example, m R 3 It is substituted with a base (for example, m is between 0 and 4).

[0022] A particular embodiment is a compound having the structure of formula (IA) (or containing a radical of the structure of formula (IA):

[0023] [ka] or a pharmaceutically acceptable salt thereof (in the formula, R 1 is optionally substituted C3-C8 alkyl (e.g., acyclic or cyclic) (e.g., optionally substituted with one or more substituents, each substituent independently being halogen, alkyl, heteroalkyl, alkoxy, or cycloalkyl); R 2 H, -OR z , -SO2N(R z )2, -NR 2x R 2y , or N3; Y is H, -OR z , -NR 2x R 2y , -SR z -SOR z , -SO3R z -N3, -COR z ,-COOR z, -CON(R z )2, -COSR z , -SO2N(R z )2, or -CON(R z )2, R 2x and R 2y These are, independently, hydrogen and -N(R). z )2, -CON(R z )2, -C(R z )2-N(R z )2, -CS-N(R z )2, or optionally substituted alkyl (for example, optionally substituted with one or more substituents, each substituent independently being oxo, halogen, alkyl, heteroalkyl, alkoxy, or cycloalkyl), and each R z These are independently hydrogen, halogens, or optionally substituted alkyls, or R 2x and R 2y They combine to form optionally substituted heterocycloalkyls (for example, optionally substituted heterocycloalkyls are monocyclic or bicyclic heterocycloalkyls, and / or optionally substituted heterocycloalkyls are 3- to 10-membered heterocycloalkyls); Each R 3 These are, independently, halogen, -N3, -CN, -NO2, and -COR z ,-COOR z , -CON(R z )2, -COSR z , -SO2N(R z )2, -CON(R z )2, alkyl, heteroalkyl, cycloalkyl, heterocycloalkyl, alkoxy, amino, hydroxy, or thiol, where alkyl, alkoxy, heteroalkyl, cycloalkyl, or heterocycloalkyl is optionally substituted; R 4 and R 5 Each of these is independently an alkyl, alkoxy, halogen, or cycloalkyl group, and the alkyl, alkoxy, or cycloalkyl group may be substituted; n is between 1 and 6; m is between 0 and 4. To provide.

[0024] In some embodiments, R of formula (I) or (IA) 1 R is a C3-C6 alkyl group which may be substituted in some cases. In one embodiment, R of formula (I) or (IA) 1 These are acyclic C3-C6 alkyl groups that may be substituted in some cases.

[0025] In some embodiments of formula (I) and formula (IA), R 2 ga-NR 2x R 2y In one embodiment of formula (I) or (IA), R 2 This is NH2.

[0026] In some embodiments, the compound is of formula:

[0027] [ka] A radical represented by (or containing) any one or more of the following, or a pharmaceutically acceptable salt thereof.

[0028] In some embodiments, n is 1 to 3. In other embodiments, n is 1 or 2. In a particular embodiment of this compound, n is 1.

[0029] One embodiment provides a compound having the structure of formula (I) or (IA), or a pharmaceutically acceptable salt thereof, where Y is -OH, OCH3, -NH2, -NHNH2, -NHCONH2, -SH, -SO2NH2, -N3, -COOH, -COCH3, -COOCH3, or -CONH. In some embodiments, Y is OH. In other embodiments, Y is NH2.

[0030] In some embodiments, the compound is of formula:

[0031] [ka] A radical represented by (or containing) any one or more of the following, or a pharmaceutically acceptable salt thereof.

[0032] In some embodiments of formula (I) or (IA), R 4 and R 5 Each of these is an alkyl group. In certain embodiments of formula (I) or (IA), R 4 and R 5 However, each is independently a C1-C4 alkyl group. In one embodiment, R 4 and R 5 Each of these is methyl.

[0033] In some embodiments, the compound is of the following formula:

[0034] [ka] It is represented by (or contains) any one of the radicals, or a pharmaceutically acceptable salt thereof.

[0035] In some embodiments, X 1 , X 2 , and X 3 Each of these is N.

[0036] In certain embodiments of formula (I) or (IA), the compound is of formula:

[0037] [ka] or a pharmaceutically acceptable salt thereof.

[0038] One embodiment is a compound represented by the structure of formula (II):

[0039] [ka] or a pharmaceutically acceptable salt thereof (in the formula, R 1 and R 3 and R 4 and R 5 are each independently H, alkyl, alkoxyl, alkenyl, alkynyl, cycloaliphatic, aryl, biaryl, halo, heteroaryl, -COR 2x ,

[0040] [Chemical formula] and; R 2 is H, -OH, -NH2, -NHR 2x , N3, -NH-CH2-NH2, -CONH2, -SO2NH2, -NH-CS-NH2,

[0041] [Chemical formula] and; Z is a group of the formula G-L-, G-O-, G-L-O-, G-L-O-alkyl-, G-L-S-, G-SO2-NH-, G-L-NR a R b -, G-L-S(O) x -alkyl-, G-L-CO-, G-L-aryl-, G-L-NH-CO-NH-, G-L-NH-O-, G-L-NH-NH-, G-L-NH-CS-NH, G-L-C(O)-alkyl-, G-L-SO2,

[0042] [Chemical formula] and; L is a linker and G is a folate receptor binding ligand; R a and R b are each independently H, halo, hydroxy, alkoxy, aryl, amino, acyl or C(O)R c and R c is alkyl, aryl, oxy or alkoxy; x is 0 to 3; R 2xand R 2y Each of these is independently selected from the group consisting of H, -OH, -CH2-OH, -NH2, -CH2-NH2, -COOMe, -COOH, -CONH2, -COCH3, alkyl, alkenyl, alkynyl, alicyclic, aryl, biaryl, and heteroaryl; R 2z Each of these is -NH2, -NR 2q R 2q’ , -OR 2q , -SO-R 2q , and -COR 2q Independently selected from the group consisting of; Each R 2q and R 2q’ These are independently alkyl or H,

[0043] [ka] It is a 3-10 member nitrogen-containing non-aromatic monocyclic or bicyclic heterocyclic; R 21 is H or alkyl; n' is between 0 and 30; In formula II, X 1 , X 2 , and X 3 Each of them operates independently, CR q or N, and each R q These are independently hydrogen, halogens, or optionally substituted alkyl groups; n is between 0 and 30 (for example, 1 to 8 or 1 to 6), and m is between 0 and 4; If n is 0, Z is not bonded to equation (II) by an oxygen atom. To provide.

[0044] One embodiment is a compound represented by the structure of formula (IIA):

[0045] [ka] or a pharmaceutically acceptable salt thereof (in the formula, R1 is optionally substituted alkyl (e.g., acyclic or cyclic) (e.g., optionally substituted with one or more substituents, each substituent independently being a halogen, alkyl, heteroalkyl, alkoxy, or cycloalkyl); R 2 H, -OR z , -SO2N(R z )2, -NR 2x R 2y , or N3, R 2x and R 2y These are, independently, hydrogen and -N(R). z )2, -CON(R z )2, -C(R z )2-N(R z )2, -CS-N(R z )2, or optionally substituted alkyl (for example, optionally substituted with one or more substituents, each substituent independently being oxo, halogen, alkyl, heteroalkyl, alkoxy, or cycloalkyl), and each R z is independently hydrogen, halogen, or optionally substituted alkyl; or R 2x and R 2y They combine to form optionally substituted heterocycloalkyls (for example, optionally substituted heterocycloalkyls are monocyclic or bicyclic heterocycloalkyls, and / or optionally substituted heterocycloalkyls are 3- to 10-membered heterocycloalkyls); Each R 3 These are independently halogens, -N3, -CN, -NO2, alkyl, heteroalkyl, cycloalkyl, heterocycloalkyl, alkoxy, aryl, heteroaryl, heterocycloalkyl, amino, hydroxy, carboxyl, or thiol, where alkyl, alkoxy, heteroalkyl, cycloalkyl, or heterocycloalkyl are optionally substituted; R 4 and R 5Each of these is independently an alkyl, alkoxy, halogen, or cycloalkyl, and the alkyl, alkoxy, and cycloalkyl are optionally substituted; each X 1 , X 2 , and X 3 CR is independent. q or N, and each R q These are independently hydrogen, halogens, or optionally substituted alkyl groups; Z is LG, L is the linker, and G is the folate receptor binding ligand; n is between 1 and 6; m is between 0 and 4. To provide.

[0046] One embodiment is a compound represented by the structure of formula (III):

[0047] [ka] or a pharmaceutically acceptable salt thereof (in the formula, R 1 , R 3 , R 4 , R 5 These are, independently, H, alkyl, alkoxyl, alkenyl, alkynyl, alicyclic, aryl, biaryl, halo, heteroaryl, and -COR. 2x ,

[0048] [ka] And R 2x and R 2y Each of these is independently selected from the group consisting of H, -OH, -CH2-OH, -NH2, -CH2-NH2, -COOMe, -COOH, -CONH2, -COCH3, alkyl, alkenyl, alkynyl, alicyclic, aryl, biaryl, and heteroaryl, R 2z Each of these is -NH2, -NR 2q R 2q’ , -OR 2q , -SO-R 2q, and -COR 2q Independently selected from the group consisting of, each R 2q and R 2q’ These are independently alkyl or H,

[0049] [ka] It is a 3-10 member nitrogen-containing non-aromatic monocyclic or bicyclic heterocyclic, and R 21 is H or alkyl, and n' is between 0 and 30; Z is the group of the formula GL-, GL-CO-, GLC(O)-alkyl-, L is the linker, and G is the folate receptor binding ligand; X 1 , X 2 , and X 3 Each of them operates independently, CR q or N, and each R q These are independently hydrogen, halogens, or optionally substituted alkyl groups; Y is as shown in formula I or IA; n is between 0 and 30; m is between 0 and 4. To provide.

[0050] In one embodiment, the compound represented by the structure of formula (IIIA):

[0051] [ka] or a pharmaceutically acceptable salt thereof (in the formula, R 1 is optionally substituted alkyl (e.g., acyclic or cyclic) (e.g., optionally substituted with one or more substituents, each substituent independently being a halogen, alkyl, heteroalkyl, alkoxy, or cycloalkyl); Y is H, -OR z , -NR 2x R 2y , -SR z -SOR z , -SO3Rz -N3, -COR z ,-COOR z ,-CONR z 2. -COSR z , -SO2N(R z )2, or -CON(R z )2, R 2x and R 2y These are, independently, hydrogen and N(R). z )2, -CON(R z )2, -C(R z )2-N(R z )2, -CS-N(R z )2, or optionally substituted alkyl (for example, optionally substituted with one or more substituents, each substituent independently being oxo, halogen, alkyl, heteroalkyl, alkoxy, or cycloalkyl), and each R z is independently hydrogen, halogen, or optionally substituted alkyl; or R 2x and R 2y They combine to form optionally substituted heterocycloalkyls (for example, optionally substituted heterocycloalkyls are monocyclic or bicyclic heterocycloalkyls, and / or optionally substituted heterocycloalkyls are 3- to 10-membered heterocycloalkyls); Each R 3 These are independently halogens, -N3, -CN, -NO2, alkyl, heteroalkyl, cycloalkyl, heterocycloalkyl, alkoxy, aryl, heteroaryl, heterocycloalkyl, amino, hydroxy, carbonyl, or thiol, where alkyl, alkoxy, heteroalkyl, cycloalkyl, or heterocycloalkyl may be substituted; R 4 and R 5 Each of these is independently an alkyl, alkoxy, halogen, or cycloalkyl, and the alkyl, alkoxy, and cycloalkyl are optionally substituted; each X 1 , X 2, and X 3 CR is independent. q or N, and each R q These are independently hydrogen, halogens, or optionally substituted alkyl groups; Z is LG, L is the linker, and G is the folate receptor binding ligand; n is between 1 and 6; m is between 0 and 4. To provide.

[0052] In some embodiments of formula (II), (IIA), (III), or (IIIA), X 1 , X 2 , and X 3 Each of these is N.

[0053] In some embodiments of formulas (II), (IIA), (III), or (IIIA), n is 1.

[0054] In some embodiments, the compound is of formula:

[0055] [ka] (In the formula, Z is LG, L is the linker, and G is the folate receptor-binding ligand.) It is represented by any one of the following, or a pharmaceutically acceptable salt thereof.

[0056] In some embodiments, the compound is the compound of formula (IIA):

[0057] [ka] (In the formula, R 1 It is a C1-C6 alkyl group optionally substituted with 1-3 substituents, each substituent independently being a halogen or a C1-C6 alkoxy; R 2 -NR 2x R 2y And R 2xand R 2y Each of these is independently either hydrogen or a C1-C6 alkyl group; Each R 3 These are independently halogens, -CN, C1-C6 alkyl, C1-C6 heteroalkyl, C3-C7 cycloalkyl, C1-C6 alkoxy, amino, hydroxy, carboxyl, or thiol; R 4 and R 5 Each of these is independently a C1-C6 alkyl group; each X 1 , X 2 , and X 3 is N; Z is GL- or GLO-, L is the linker, and G is the folate receptor binding ligand; n is 1; m is between 0 and 4. or a pharmaceutically acceptable salt thereof.

[0058] In some embodiments, R 1 is C1-C6 alkyl. In some embodiments, R 2 is -NH2. In some embodiments, m is 0. In certain embodiments, R 1 The C1-C6 alkyl group is R 2 is -NH2, n is 1, and m is 0.

[0059] In some embodiments, the compound of formula (II) is the compound of formula (IIB):

[0060] [ka] That is the case.

[0061] In some embodiments, the compound has the structure:

[0062] [ka] (In the formula, Z is LG, L is the linker, and G is the folate receptor-binding ligand.) It is represented by one or more of the following, or a pharmaceutically acceptable salt thereof.

[0063] In some embodiments of formulas (II), (IIA), (IIB), (III), or (IIIA), L is a cleavable (release) linker. In specific embodiments of formulas (II), (IIA), (IIB), (III), or (IIIA), L is a hydrolyzable linker. In preferred embodiments, L is a non-cleavable or non-release linker. In specific embodiments of formulas (II), (IIA), (IIB), (III), or (IIIA), L is a non-hydrolyzable linker.

[0064] In some embodiments, L may include a substituted heteroalkyl group. In some embodiments, the heteroalkyl group is unsubstituted. In other embodiments, the heteroalkyl group is substituted with at least one substituent selected from the group consisting of alkyl, hydroxyl, acyl, polyethylene glycol (PEG), carboxylate, and halo. In yet another embodiment, L includes a substituted heteroalkyl group having at least one disulfide bond in its skeleton.

[0065] In some embodiments, L is a peptide or peptidoglycan having at least one disulfide bond in its backbone.

[0066] In some embodiments, L is a cleavable / releasing linker that can be cleaved by enzymatic reactions, reactive oxygen species (ROS), or reducing conditions.

[0067] In some embodiments, L is the formula -NH-CH2-CR 6 R 7 -SS-CH2-CH2-O-CO-(wherein, R 6 and R 7 Each of these elements independently comprises H, alkyl, or heteroalkyl. In some preferred embodiments, L does not contain a disulfide.

[0068] In some embodiments, L is given by the formula:

[0069] [ka] (In the formula, p is an integer between 0 and 30; d is an integer between 1 and 40; R 8 and R 9 (Each of these is independently H, alkyl, heterocyclyl, cycloalkyl, aryl, or heteroalkyl.) It is a base of, or contains, this base.

[0070] In some embodiments of formulas (II), (IIA), (IIB), (III), or (IIIA), L is a non-ejectable linker. In some specific embodiments, L is a non-hydrolyzable linker.

[0071] In some embodiments, L is one or more base linker portions (L') (e.g., L' n’ (represented by ). In some embodiments, each of the one or more linker moieties is independently selected from the group consisting of alkylenes, heteroalkylenes, -O-alkylenes, alkenylenes, acyls, aryls, heteroaryls, amides, oximes, ethers, esters, triazoles, PEGs, carboxylates, carbonates, carbamates, amino acids, peptides (e.g., containing two or more amino acid residues), and peptidoglycans.

[0072] In one embodiment, L is an alkyl ether or contains one. In another embodiment, L is an amide or contains one. In another embodiment, L is a peptide or peptidoglycan or contains one. In another embodiment, L is an amino acid or contains one. In another embodiment, L is PEG (e.g., -OCH2-CH2-O-) or contains one. In another embodiment, L is a polysaccharide or contains one.

[0073] In some embodiments, L is structure:

[0074] [ka] (In the formula, w is between 0 and 5, and p is between 1 and 30) It is a base represented by, or includes.

[0075] In one embodiment, L is

[0076] [ka]

[0077] [ka] (In the formula, n'' is an integer between 0 and 30 (for example, 1 to 30, 1 to 8, or 1 to 6)). It is either or includes this.

[0078] In a particular embodiment, L is a divalent linker.

[0079] In some embodiments of formulas (II), (IIA), (IIB), (III), or (IIIA), G is the base of formula (IV):

[0080] [ka] (In the formula, R is as follows:

[0081] [ka] (or a naturally occurring or unnatural amino acid or its derivative or fragment, or a component thereof) It is either or contains this group.

[0082] In certain specific embodiments of formulas (II), (IIA), (IIB), (III), or (IIIA), G is the structure of formula (V):

[0083] [ka] It is a radical having (for example, being the group or containing the group).

[0084] In some specific embodiments of formulas (II), (IIA), (IIB), (III), or (IIIA), G is the structure of formula (VI):

[0085] [ka] It is a radical having (for example, being the group or containing the group).

[0086] In some embodiments, the compound has the following structure:

[0087] [ka] It is represented by one of the following, or a pharmaceutically acceptable salt thereof (wherein n1 is between 0 and 10, and n2 is between 0 and 10).

[0088] In some embodiments, the compound has the following structure:

[0089] [ka]

[0090] [ka] It is represented by one of the following.

[0091] In some embodiments, the compound has the following structure:

[0092] [ka] It is represented by one of the following.

[0093] In a particular embodiment, the compound has the following structure:

[0094] [ka]

[0095] [ka]

[0096] [ka] It is represented by one of the following.

[0097] In a particular embodiment, the compound has the following structure:

[0098] [ka] It is represented by one of the following.

[0099] A pharmaceutical composition is provided comprising a compound of formula (I), (IA), (II), (IIA), (IIB), (III), or (IIIA), or any compound encompassed by such formulas, or a pharmaceutically acceptable salt thereof, and at least one pharmaceutically acceptable excipient.

[0100] Also provided are pharmaceutical compositions comprising a therapeutically effective compound of formula (I), (IA), (II), (IIA), (IIB), (III), or (IIIA), or any compound encompassed by such formulas, or a pharmaceutically acceptable salt thereof, and at least one pharmaceutically acceptable excipient.

[0101] Further provided are methods for treating cancer or inflammatory diseases or disorders, such as fibrotic diseases or disorders, in individuals requiring such treatment. The method comprises administering a therapeutically effective amount of one or more compounds from any one of (I), (IA), (II), (IIA), (IIB), (III), or (IIIA), or any compound encompassed by such formulas, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition containing the same, to an individual or subject requiring such treatment.

[0102] In certain embodiments of this method, which is for treating inflammatory diseases or disorders, the inflammatory disease or disorder is selected from the group consisting of lupus, inflammatory bowel disease (IBS), Addison's disease, Graves' disease, Sjögren's syndrome, celiac disease, Hashimoto's thyroiditis, myasthenia gravis, autoimmune vasculitis, reactive arthritis, psoriatic arthritis, pernicious anemia, ulcerative colitis, rheumatoid arthritis, type 1 diabetes, multiple sclerosis, transplant rejection, fatty liver disease, asthma, osteoporosis, sarcoidosis, ischemia-reperfusion injury, prosthetic osteolysis, glomerulonephritis, scleroderma, psoriasis, autoimmune myocarditis, spinal cord injury, central nervous system disorders, viral infections, influenza, coronavirus infection, cytokine storm syndrome, bone injury, inflammatory brain disease, and atherosclerosis. In certain embodiments, the inflammatory disease or disorder is a fibrous disease or disorder.

[0103] Further methods are provided for treating fibrotic diseases or disorders in individuals in need. The method comprises administering a therapeutically effective amount of one or more compounds of any one of (I), (IA), (II), (IIA), (IIB), (III), or (IIIA), or any compound encompassed by such formulas, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition containing the same, to an individual or subject in need. In certain embodiments, the fibrotic disease or disorder is selected from the group consisting of articular fibrosis, autoimmune pancreatitis, bladder fibrosis, chronic kidney disease, chronic wound, Crohn's disease, tendonoid, Dupuytren's contracture, endometrial fibrosis, fibromatosis, graft-versus-host disease (GVHD), cardiac fibrosis, keloid, hepatic fibrosis (e.g., non-alcoholic steatohepatitis (NASH) or cirrhosis), mediastinal fibrosis, myelofibrosis, nephrogenic systemic fibrosis, Peyronie's disease, pulmonary fibrosis, retroperitoneal fibrosis, scleroderma or systemic sclerosis, and cutaneous fibrosis.

[0104] Another method provided is a method for treating cancer in an individual in need. This method comprises the step of administering (e.g., to the individual) a compound of formula (I), (IA), (II), (IIA), (IIB), (III), or (IIIA), or any compound encompassed by such formulas, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition containing the same, to an individual in need. In one embodiment, cancer is defined as lung cancer, bone cancer, pancreatic cancer, skin cancer, head cancer, cervical cancer, cutaneous melanoma, intraocular melanoma, uterine cancer, ovarian cancer, endometrial cancer, epithelial cancer, leiomyosarcoma, rectal cancer, stomach cancer, colon cancer, breast cancer, triple-negative breast cancer, fallopian tube cancer, endometrial cancer, cervical cancer, vaginal cancer, vulvar cancer, Hodgkin's disease, esophageal cancer, small intestine cancer, endocrine cancer, thyroid cancer, parathyroid cancer, non-small cell lung cancer, small cell lung cancer, adrenal cancer, soft tissue sarcoma, urethral cancer, penile cancer, prostate cancer, chronic leukemia, acute leukemia, lymphocytic lymphoma, pleural mesothelioma, bladder cancer, gastric cancer. The cancer is selected from the group consisting of Burkitt lymphoma, ureteral cancer, kidney cancer, renal cell carcinoma, renal pelvis cancer, central nervous system (CNS) neoplasms, primary CNS lymphoma, vertebral axial tumor, brainstem glioma, pituitary adenoma, cholangiocarcinoma, Haasle cell thyroid cancer, and gastroesophageal junction adenocarcinoma. In additional embodiments, the cancer is lung cancer, breast cancer (e.g., triple-negative breast cancer), colon cancer, gastric cancer, bladder cancer, ovarian cancer, pancreatic cancer, or epithelial cancer.

[0105] Another method provided is a method for inhibiting or reducing fibrosis (for example, in an individual in need, e.g., an individual suffering from cancer or a fibrotic disease). The method comprises the step of administering a compound of formula (I), (IA), (II), (IIA), (IIB), (III), or (IIIA) or any compound encompassed by such formulas, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition containing the same, to an individual in need (e.g., to the individual). In some embodiments, the fibrotic disease or disorder is selected from the group consisting of articular fibrosis, autoimmune pancreatitis, bladder fibrosis, chronic kidney disease, chronic wound, Crohn's disease, tendonoid, Dupuytren's contracture, endometrial fibroma, fibromatosis, graft-versus-host disease, cardiac fibrosis, keloid, hepatic fibrosis (e.g., NASH or cirrhosis), mediastinal fibrosis, myelofibrosis, nephrogenic systemic fibrosis, Peyronie's disease, pulmonary fibrosis, retroperitoneal fibrosis, scleroderma or systemic sclerosis, and cutaneous fibrosis.

[0106] In one embodiment, the fibrotic disease or disorder is idiopathic pulmonary fibrosis, hepatic fibrosis, myelofibrosis, or cardiac fibrosis. In a particular embodiment, the fibrotic disease or disorder is pulmonary fibrosis, hepatic fibrosis, scleroderma, myelofibrosis, Crohn's disease, or chronic kidney disease.

[0107] A method is provided for inhibiting or reducing fibrosis (for example, in an individual in need, e.g., an individual suffering from cancer or a fibrotic disease), comprising the step of administering to an individual in need (e.g., to the individual) an amount effective in converting a population of macrophages biased toward an M2-like phenotype (e.g., pro-fibrotic) to an M1-like phenotype (e.g., anti-fibrotic), wherein the population of macrophages is located at a targeted site within the individual, the M2-like phenotype is associated with an anti-inflammatory / pro-fibrotic state, and the M1-like phenotype is associated with an anti-inflammatory / anti-fibrotic state.

[0108] Furthermore, a method is provided for inhibiting or reducing cancerous growth (for example, in an individual in need, e.g., an individual with cancer), comprising the step of administering to an individual in need (e.g., to the individual) an amount effective in converting a population of macrophages biased toward an M2-like phenotype (e.g., pro-fibrotic) to an M1-like phenotype (e.g., anti-fibrotic), wherein the population of macrophages is located at a target site within the individual, the M2-like phenotype is associated with an anti-inflammatory / pro-fibrotic state, and the M1-like phenotype is associated with an anti-inflammatory / anti-fibrotic state. In at least one embodiment, the target site is the tumor microenvironment.

[0109] In any of the above embodiments, this method does not induce unnecessary inflammation in the individual.

[0110] In any of the above embodiments, the method further includes the step of administering a second therapeutic agent. In one embodiment, the second therapeutic agent is an anti-inflammatory agent. In any of the above embodiments, the method further includes the step of administering a chemotherapeutic agent.

[0111] Further embodiments and the full scope of applicability of this disclosure will become apparent from the detailed description. However, it should be understood that the detailed description and specific examples are given for illustrative purposes only. Various changes and modifications within the spirit and scope of this disclosure will become apparent to those skilled in the art.

[0112] A more detailed explanation will be better understood when viewed in conjunction with the accompanying diagrams. [Brief explanation of the drawing]

[0113] [Figure 1] This figure shows the chemical structures of the exemplary compounds provided herein. [Figure 2]This figure shows the effects of various exemplary compounds on interleukin-6 (IL-6) expression in peripheral blood mononuclear cells (PBMCs). [Figure 3-1] This figure shows the in vitro effects of various exemplary compounds on IL-6 (Figure 3A) and CXCL-10 (Figure 3B) induction in human monocyte-derived M2 macrophages over 48 hours. [Figure 3-2] This figure shows the in vivo effects of various exemplary compounds on IL-6 (Figure 3C) and tumor necrosis factor alpha (TNF-α) (Figure 3D) production in mice. [Figure 4] This figure shows the effects of Toll-like receptor 7 (TLR7) agonists on IL-6 and TNF-α in mouse bone marrow-derived M2 macrophages. [Figure 5] This figure shows flow cytometry data of TLR7 and folate receptor β (FR-β) expression in fixed and permeabilized human M2 polarized macrophages. [Figure 6] This figure shows confocal microscopy images of human PBMC-derived M2 macrophages that were fixed, permeabilized, and stained with both TLR7 antibody and FR-β antibody (the results indicate that both antibodies are located within endosomes). [Figure 7-1] This figure shows liquid chromatography-mass spectrometry (LCMS) data from disulfide cleavage tests of folic acid (FA)-TLR7 conjugate FA-TLR7-1 (Figure 7A) and FA-compound 1 (releasing conjugate) (compound 5) (Figure 7B), performed at 0, 7, 30, and 50 minutes. [Figure 7-2] This figure shows liquid chromatography-mass spectrometry (LCMS) data from disulfide cleavage tests of folic acid (FA)-TLR7 conjugate FA-TLR7-1 (Figure 7A) and FA-compound 1 (releasing conjugate) (compound 5) (Figure 7B), performed at 0, 7, 30, and 50 minutes. [Figure 8]This figure shows schematic diagrams of possible mechanisms of action for certain releasing folate-TLR7 conjugates and non-releasing folate-TLR7 conjugates, based on data obtained to date. [Figure 9] This figure shows graphic data representing the expression of the M1 marker IL-6 in PBMC-derived macrophages after treatment with a releasing folate-TLR7 conjugate (FA-PEG3-TLR7-1A (releasing ("Re"))). [Figure 10-1] This figure shows graphical data from an in vivo treatment study of a releasing folate-TLR7 conjugate (compound 5; FA-PEG3-TLR7-1A(Re)) (10 nmol / mouse) in a 4T1 solid tumor model. Figure 10A shows tumor volume measured every other day after treatment. Figure 10B shows the average tumor weight measured at the end of the study. Figure 10C shows the relative ratio of M1 / M2 (CD86+ / CD206+) macrophages. Figure 10D shows CD8+ cells per 100,000 events. Figure 10E shows CD4+ cells per 100,000 events. [Figure 10-2] This figure shows graphical data from an in vivo treatment study of a releasing folate-TLR7 conjugate (compound 5; FA-PEG3-TLR7-1A(Re)) (10 nmol / mouse) in a 4T1 solid tumor model. Figure 10A shows tumor volume measured every other day after treatment. Figure 10B shows the average tumor weight measured at the end of the study. Figure 10C shows the relative ratio of M1 / M2 (CD86+ / CD206+) macrophages. Figure 10D shows CD8+ cells per 100,000 events. Figure 10E shows CD4+ cells per 100,000 events. [Figure 11]This figure shows graphical data on various M1 markers in human PBMC-derived macrophages after treatment with the non-releasing folate-TLR7 conjugate (compound 4; FA-PEG3-TLR7-1A(NR)) of the present disclosure. Figure 11A shows the change in mRNA levels of the M1 marker IL-6 after 3 hours of treatment, and Figure 11B shows the change in mRNA levels of the M1 marker TNF-α after 3 hours of treatment. Figure 11C shows the analysis of IL-6 protein expression using ELISA after 3 + 45 hours of treatment. Figure 11D shows the surface expression of the M2 marker CD206 in macrophages analyzed by flow cytometry. [Figure 12] This figure shows graphical data regarding the cell surface markers CD40 and CD80, respectively, analyzed by flow cytometry in human PBMC-derived M2 macrophages after treatment with a non-releasing folate-TLR7 conjugate (compound 4; FAPEG3-TLR7-1A(NR)). [Figure 13] This figure shows in vivo pharmacokinetic analysis data of a non-releasing folate-TLR7 conjugate (compound 4; FA-PEG3-TLR7-1A(NR)) in mice (column: Agilent Eclipse Plus C18, 2.1 × 50 mm, SN: B17477, eluent: A - water + 0.1% FA, B - CAN + 0.1% FA). [Figure 14] This figure shows graphical data from in vivo treatment trials of non-releasing folate-TLR7 conjugate (compound 4; FA-PEG3-TLR7-1A(NR)) at different concentrations and dosing intervals in a 4T1 solid tumor model. Figure 14A shows the tumor volume measured after treatment. Figure 14B shows the tumor weight (grams) measured at the end of the trial. [Figure 15]This figure shows data from an in vivo treatment trial of a non-releasing folate-TLR7 conjugate (compound 4; FA-PEG3-TLR7-1A(NR)) (1 nmol once / week) in a 4T1 metastatic tumor model. Figure 15A shows the tumor volume measured during treatment, Figure 15B shows the quantification of metastatic tumor cells in the lung using a 6-thioguanine assay, and Figure 15C shows a representative image from the 6-thioguanine assay in Figure 15B. [Modes for carrying out the invention]

[0114] This disclosure relates to the preparation and use of compounds and compositions for the prevention and / or treatment of fibrotic diseases. In certain embodiments, the compounds, compositions, and methods provided are also useful for the prevention and / or treatment of cancer. In some embodiments, the compounds, compositions, and methods provided utilize strategies that (e.g., selectively) target the innate immune system and reprogram macrophage polarization from M2 to M1, for example, by utilizing the anti-fibrotic / pro-inflammatory properties of the M1 phenotype.

[0115] In at least one embodiment, such compounds and compositions include an immunomodulator or a pharmaceutically acceptable salt thereof (e.g., a Toll-like receptor (TLR) agonist, e.g., a TLR7 or TLR7 / 8 agonist) that, upon administration, can convert activated myeloid cells (e.g., M2-like macrophages) to anti-fibrotic / pro-inflammatory M1 polarization—e.g., reprogram them).

[0116] In exemplary embodiments, an immunomodulator or a pharmaceutically acceptable salt thereof is conjugated (directly or via a linker) to a targeting moiety or radical that targets pattern recognition receptors on fibrous or cancerous cells. The targeting moiety or radical may be, for example, a folate ligand or a functional fragment or analogue thereof. Such embodiments utilize the restricted expression of the target of the targeting moiety to directly localize a systemically administered compound to target-expressing cells (e.g., cells in fibrous and / or cancerous tissues), allowing the immunomodulator to reprogram activated myeloid cells (e.g., M2-like macrophages) to anti-fibrotic / pro-inflammatory M1 polarization. This targeting design advantageously prevents systemic activation of the immune system and thus avoids toxicity in the target.

[0117] Furthermore, exemplary embodiments may include a linker positioned between the targeting moiety and the immunomodulator or a pharmaceutically acceptable salt thereof. Such a linker may be release-oriented or non-release-oriented. Compounds containing a release-oriented linker (and compositions containing the compound), upon administration, cause the targeting moiety and the immunomodulator to release from each other at or around the time the immunomodulator becomes active. In embodiments in which compounds containing a non-release-oriented linker (and compositions containing the compound) are administered, the targeting moiety and the immunomodulator do not rapidly release under physiological conditions. Thus, the components remain together after uptake by targeted cells (e.g., fibrous or cancerous cells) and / or activation of the immunomodulator.

[0118] Here, we present data relating to various embodiments and supporting examples.

[0119] compound One embodiment is a compound represented by the structure of formula (I) (e.g., an immunomodulator):

[0120] [ka] or a pharmaceutically acceptable salt thereof (in the formula, R 1 , R 3 , R 4 , R 5 These are, independently, H, alkyl, alkoxyl, alkenyl, alkynyl, cycloalkyl, alicyclic, aryl (e.g., biaryl), halo, heteroaryl, and -COR. 2x ,

[0121] [ka] and; R 2 H, -OH, -NH2, -NHR 2x , N3, -NH-CH2-NH2, -CONH2, -SO2NH2, -NH-CS-NH2,

[0122] [ka] and; Y is H, -OH, -NH2, -NHR 2x , -OR 2x , -SO-R 2x , -SH, -SO3H, -N3, -CHO, -COOH, -CONH2, -COSH, -COR 2x -SO2NH2, alkenyl, alkynyl, alkoxyl, -NH-CH2-NH2, -CONH2, -SO2NH2, -NH-CS-NH2,

[0123] [ka] And R 2x and R 2y Each of these is independently selected from the group consisting of H, -OH, -CH2-OH, -NH2, -CH2-NH2, -COOMe, -COOH, -CONH2, -COCH3, alkyl, alkenyl, alkynyl, alicyclic, aryl, biaryl, and heteroaryl. R 2z Each of these is -NH2, -NR 2q R 2q’, -OR 2q , -SO-R 2q , and -COR 2q Independently selected from the group consisting of; each R 2q and R 2q’ These are independently alkyl or H,

[0124] [ka] It is a 3-10 member nitrogen-containing non-aromatic monocyclic or bicyclic heterocyclic; R 21 is H or alkyl; n' is between 0 and 30; In formula (I), X 1 , X 2 , X 3 Each of these is independent of CR q or N, and each R q These are independently hydrogen, halogens, or optionally substituted alkyl groups; n is between 0 and 30; m is between 0 and 4. To provide.

[0125] In at least one embodiment of formula (I), when n is 0, Y is either H, -OH, or -OR 2x But no.

[0126] Another embodiment involves a compound having the structure of formula (IA) (e.g., an immunomodulator):

[0127] [ka] or a pharmaceutically acceptable salt thereof (in the formula, R 1 is optionally substituted C3-C8 alkyl (e.g., acyclic or cyclic) (e.g., optionally substituted with one or more substituents, each substituent independently being halogen, alkyl, heteroalkyl, alkoxy, or cycloalkyl); R 2 H, -ORz , -SO2N(R z )2, -NR 2x R 2y , or N3; Y is H, -OR z , -NR 2x R 2y , -SR z -SOR z , -SO3R z -N3, -COR z ,-COOR z , -CON(R z )2, -COSR z , -SO2N(R z )2, or -CON(R z )2, R 2x and R 2y These are, independently, hydrogen and -N(R). z )2, -CON(R z )2, -C(R z )2-N(R z )2, -CS-N(R z )2, or optionally substituted alkyl (for example, optionally substituted with one or more substituents, each substituent independently being oxo, halogen, alkyl, heteroalkyl, alkoxy, or cycloalkyl), and each R z is independently hydrogen, halogen, or optionally substituted alkyl; or R 2x and R 2y They combine to form optionally substituted heterocycloalkyls (for example, optionally substituted heterocycloalkyls are monocyclic or bicyclic heterocycloalkyls, and / or optionally substituted heterocycloalkyls are 3- to 10-membered heterocycloalkyls); Each R 3 These are, independently, halogen, -N3, -CN, -NO2, and -COR z ,-COOR z , -CON(R z )2, -COSR z , -SO2N(R z )2, or -CON(R z)2, alkyl, heteroalkyl, cycloalkyl, heterocycloalkyl, alkoxy, amino, hydroxy, or thiol, where alkyl, alkoxy, heteroalkyl, cycloalkyl, or heterocycloalkyl is optionally substituted; R 4 and R 5 Each of these is independently an alkyl, alkoxy, halogen, or cycloalkyl, and the alkyl, alkoxy, and cycloalkyl are optionally substituted; n is between 1 and 6; m is between 0 and 4. To provide.

[0128] One embodiment provides a compound having the structure of formula (I) or (IA), where n is 1 to 30, or a pharmaceutically acceptable salt thereof. In one embodiment, n is 1 to 6. In another embodiment, n is 1 to 3. In another embodiment, n is 1 or 2. In another embodiment, n is 0. In another embodiment, n is 1. In another embodiment, n is 1 and Y is OH. In another embodiment, n is 1 and Y is NH2. In one embodiment, the compound has the structure of compound 1:

[0129] [ka] It is represented by or a pharmaceutically acceptable salt thereof.

[0130] In one embodiment, the compound is represented by the structure of compound 2. In another embodiment, the compound is represented by the structure of compound 3. The structures of such compounds are shown in Figure 1.

[0131] One embodiment provides a compound having the structure of formula (I) or (IA), or a pharmaceutically acceptable salt thereof, wherein Y is -OH, OCH3, -NH2, -NHNH2, -NHCONH2, -SH, -SO2NH2, -N3, -COOH, -COCH3, -COOCH3, or -CONH2.

[0132] In one embodiment, Y is H, -NH2, -NHR 2x , -OR 2X , -SO-R 2x , -SH, -SO3H, -N3, -CHO, -COOH, -CONH2, -COSH, -COR 2x -SO2NH2, alkenyl, alkynyl, alkoxyl, -NH-CH2-NH2, -CONH2, -SO2NH2, -NH-CS-NH2,

[0133] [ka] The present invention provides a compound having the structure of formula (I) or (IA), or a pharmaceutically acceptable salt thereof.

[0134] One embodiment provides a compound having the structure of formula (I) or (IA), where Y is OH, or a pharmaceutically acceptable salt thereof.

[0135] One embodiment provides a compound having the structure of formula (I) or (IA), where Y is NH2, or a pharmaceutically acceptable salt thereof.

[0136] One embodiment provides a compound having the structure of formula (I) or (IA), where n is 1 and Y is OH, or a pharmaceutically acceptable salt thereof.

[0137] One embodiment provides a compound having the structure of formula (I) or (IA), where n is 1 and Y is NH2, or a pharmaceutically acceptable salt thereof.

[0138] One embodiment provides a compound having the structure of formula (I) or (IA), where n is 0 and Y is NH2, or a pharmaceutically acceptable salt thereof.

[0139] One embodiment is R 1 The present invention provides a compound having the structure of formula (I) or (IA), wherein R is optionally substituted with an alkyl group, or a pharmaceutically acceptable salt thereof. In one embodiment, R1 In some embodiments, R is a substituted C3-C6 alkyl group. 1 R is an acyclic C3-C6 alkyl group which may be substituted. In another embodiment, R 1 It is butyl.

[0140] One embodiment is R 2 ga-NR 2x R 2y The present invention provides a compound having the structure of formula (I) or (IA), or a pharmaceutically acceptable salt thereof. In one embodiment, R 2 This is NH2.

[0141] One embodiment is R 3 The present invention provides a compound having the structure of formula (I) or (IA), wherein the element is H, or a pharmaceutically acceptable salt thereof.

[0142] One embodiment is R 4 The present invention provides a compound having the structure of formula (I) or (IA), wherein R is alkyl, or a pharmaceutically acceptable salt thereof. In one embodiment, R 4 It is methyl.

[0143] One embodiment is R 5 The present invention provides a compound having the structure of formula (I) or (IA), wherein R is alkyl, or a pharmaceutically acceptable salt thereof. In one embodiment, R 5 It is methyl.

[0144] One embodiment is R 4 and R 5 The present invention provides a compound having the structure of formula (I) or (IA), wherein each of the elements is alkyl, or a pharmaceutically acceptable salt thereof. In one embodiment, R 4 and R 5 However, each is independently a C1-C4 alkyl group. In one embodiment, R 4 and R 5 Each of these is methyl.

[0145] One embodiment provides a compound having the structure of formula (I) or (IA), or a pharmaceutically acceptable salt thereof, where m is 0. In another embodiment, m is 1. In another embodiment, m is 2. In another embodiment, m is 3. In another embodiment, m is 4.

[0146] One embodiment is X 1 , X 2 , and X 3 The present invention provides a compound having the structure of formula (I), wherein each of the elements is N, or a pharmaceutically acceptable salt thereof. In one embodiment, X 1 In another embodiment, X 2 In another embodiment, X 3 This is N.

[0147] One embodiment provides a compound having the structure of formula (I), or a pharmaceutically acceptable salt thereof, excluding compounds in which n is 0.

[0148] One embodiment provides a compound having the structure of formula (I), or a pharmaceutically acceptable salt thereof, excluding compounds in which n is 0 and Y is OH.

[0149] One embodiment provides a compound having the structure of formula (I), or a pharmaceutically acceptable salt thereof, wherein n is 0, Y is OH, and R 1 is butyl, and R 2 NH2 is R 3 H is R 4 and R 5 Exclude compounds where each of the two atoms is methyl.

[0150] One embodiment provides a compound having the structure of formula (I), or a pharmaceutically acceptable salt thereof, excluding the compound TLR7-1 shown in Figure 1.

[0151] In some embodiments, the compound is of formula:

[0152] [ka] It is represented by one or more of the following, or a pharmaceutically acceptable salt thereof.

[0153] In some embodiments, the compound is of formula:

[0154] [ka] It is represented by one or more of the following, or a pharmaceutically acceptable salt thereof.

[0155] In some embodiments, the compound is of formula:

[0156] [ka] It is represented by one or more of the following, or a pharmaceutically acceptable salt thereof.

[0157] Conjugate This disclosure further relates to compounds (and their radicals) provided herein (e.g., TLR7 / 8 agonists) conjugated to a targeting moiety that targets cellular pattern recognition receptors, either directly or via a linker. In some embodiments, the targeting ligand comprises a folic acid ligand or a functional fragment or analog thereof, e.g., a pteroyl amino acid. In some embodiments, the linker is non-releasing. In some embodiments, the conjugate provides a targeting molecule having a non-releasing linker, thereby reducing systemic exposure to the TLR7 / 8 agonist. In some embodiments, the conjugate provides a targeting molecule having a non-releasing linker, thereby reducing the systemic adverse effects of the TLR7 / 8 agonist.

[0158] One embodiment is a compound represented by the structure of formula (II):

[0159] [ka] or a pharmaceutically acceptable salt thereof (in the formula, R 1 , R 3 , R 4 , R 5 These are, independently, H, alkyl, alkoxyl, alkenyl, alkynyl, alicyclic, aryl, biaryl, halo, heteroaryl, and -COR. 2x ,

[0160] [ka] and; R 2 H, -OH, -NH2, -NHR 2x , N3, -NH-CH2-NH2, -CONH2, -SO2NH2, -NH-CS-NH2,

[0161] [ka] and; Z is represented by the formulas GL-, GO-, GLO-, GLO-alkyl-, GLS-, G-SO2-NH-, and GL-NR a R b -, GLS(O) x -alkyl-, GL-CO-, GL-aryl-, GL-NH-CO-NH-, GL-NH-O-, GL-NH-NH-, GL-NH-CS-NH, GLC(O)-alkyl-, GL-SO2-,

[0162] [ka] It is the basis of, L is the linker, and G is the folate receptor binding ligand; R a and R b Each of these is independently H, halo, hydroxy, alkoxy, aryl, amino, acyl, or C(O)R c And R c These are alkyl, aryl, oxy, or alkoxy; x is 0 to 3 (for example, an integer that varies between 0 and 3); R 2x and R 2y Each of these is independently selected from the group consisting of H, -OH, -CH2-OH, -NH2, -CH2-NH2, -COOMe, -COOH, -CONH2, -COCH3, alkyl, alkenyl, alkynyl, alicyclic, aryl, biaryl, and heteroaryl; Each R 2z -NH2, -NR 2q R 2q’ , -OR 2q , -SO-R 2q , and -COR 2q Independently selected from the group consisting of, each R 2q and R 2q’ These are independently alkyl or H,

[0163] [ka] It is a 3-10 member nitrogen-containing non-aromatic monocyclic or bicyclic heterocyclic; R 21 is H or alkyl; n' is between 0 and 30; In formula II, X 1 , X 2 , X 3 Each of them operates independently, CR q or N, and each R q These are independently hydrogen, halogen, or optionally substituted alkyl groups. n is an integer between 0 and 30 (for example, an integer that varies between 0 and 30); m is between 0 and 4. To provide.

[0164] In certain embodiments of the compound of formula II, when n is 0, Z is not bonded to formula (II) by an oxygen atom.

[0165] One embodiment is a compound represented by the structure of formula (IIA):

[0166] [ka] or a pharmaceutically acceptable salt thereof (in the formula, R 1 is optionally substituted alkyl (e.g., acyclic or cyclic) (e.g., optionally substituted with one or more substituents, each substituent independently being a halogen, alkyl, heteroalkyl, alkoxy, or cycloalkyl); R 2 H, -OR z , -SO2N(R z )2, -NR 2x R 2y , or N3, R 2x and R 2y These are, independently, hydrogen and -N(R). z )2, -CON(R z )2, -C(R z )2-N(R z )2, -CS-N(R z )2, or optionally substituted alkyl (for example, optionally substituted with one or more substituents, each substituent independently being oxo, halogen, alkyl, heteroalkyl, alkoxy, or cycloalkyl), and each R z is independently hydrogen, halogen, or optionally substituted alkyl; or R 2x and R 2y They combine to form optionally substituted heterocycloalkyls (for example, optionally substituted heterocycloalkyls are monocyclic or bicyclic heterocycloalkyls, and / or optionally substituted heterocycloalkyls are 3- to 10-membered heterocycloalkyls); Each R 3These are independently halogens, -N3, -CN, -NO2, alkyl, heteroalkyl, cycloalkyl, heterocycloalkyl, alkoxy, aryl, heteroaryl, heterocycloalkyl, amino, hydroxy, carboxyl, or thiol, where alkyl, alkoxy, heteroalkyl, cycloalkyl, or heterocycloalkyl are optionally substituted; R 4 and R 5 Each of these is independently an alkyl, alkoxy, halogen, or cycloalkyl, and the alkyl, alkoxy, and cycloalkyl are optionally substituted; each X 1 , X 2 , and X 3 CR is independent. q or N, and each R q These are independently hydrogen, halogens, or optionally substituted alkyl groups; Z is LG, L is the linker, and G is the folate receptor binding ligand; n is between 1 and 6; m is between 0 and 4. To provide.

[0167] One embodiment is a compound represented by the structure of formula (IIA):

[0168] [ka] (In the formula, R 1 It is a C1-C6 alkyl group optionally substituted with 1-3 substituents, each substituent independently being a halogen or a C1-C6 alkoxy; R 2 -NR 2x R 2y And R 2x and R 2y Each of these is independently either hydrogen or a C1-C6 alkyl group; Each R 3These are independently halogens, -CN, C1-C6 alkyl, C1-C6 heteroalkyl, C3-C7 cycloalkyl, C1-C6 alkoxy, amino, hydroxy, carboxyl, or thiol; R 4 and R 5 Each of these is independently a C1-C6 alkyl group; each X 1 , X 2 , and X 3 is N; Z is GL- or GLO-, L is the linker, and G is the folate receptor binding ligand; n is 1; m is between 0 and 4. or provide a pharmaceutically acceptable salt thereof.

[0169] One embodiment has the structure of formula (IIB):

[0170] [ka] The present invention provides a compound of formula (II) having, or a pharmaceutically acceptable salt thereof.

[0171] One embodiment provides a compound having the structure of formula (II), (IIA), or (IIB), where n is 1 to 30, or a pharmaceutically acceptable salt thereof. In one embodiment, n is 1 to 6. In another embodiment, n is 1 to 3. In another embodiment, n is 1 or 2. In another embodiment, n is 0. In another embodiment, n is 1.

[0172] One embodiment is R 1 The present invention provides compounds having the structure of formula (II), (IIA), or (IIB), wherein R is optionally substituted with an alkyl group, or pharmaceutically acceptable salts thereof. In one embodiment, R 1 In some embodiments, R is a substituted C3-C6 alkyl group. 1 R is an acyclic C3-C6 alkyl group which may be substituted. In another embodiment, R 1It is butyl.

[0173] One embodiment is R 2 ga-NR 2x R 2y The present invention provides a compound having the structure of formula (II) or (IIA), or a pharmaceutically acceptable salt thereof. In one embodiment, R 2 This is NH2.

[0174] One embodiment is R 3 The present invention provides a compound having the structure of formula (II) or (IIA), wherein the element is H, or a pharmaceutically acceptable salt thereof.

[0175] One embodiment is R 4 The present invention provides a compound having the structure of formula (II) or (IIA), wherein R is alkyl, or a pharmaceutically acceptable salt thereof. In one embodiment, R 4 It is methyl.

[0176] One embodiment is R 5 The present invention provides a compound having the structure of formula (II) or (IIA), wherein R is alkyl, or a pharmaceutically acceptable salt thereof. In one embodiment, R 5 It is methyl.

[0177] One embodiment is R 4 and R 5 The present invention provides a compound having the structure of formula (II) or (IIA), wherein each of the elements is an alkyl group, or a pharmaceutically acceptable salt thereof. In one embodiment, R 4 and R 5 Each of these is methyl.

[0178] One embodiment provides a compound having the structure of formula (II) or (IIA), or a pharmaceutically acceptable salt thereof, where m is 0. In another embodiment, m is 1. In another embodiment, m is 2. In another embodiment, m is 3. In another embodiment, m is 4.

[0179] One embodiment is X 1 , X2 , and X 3 The present invention provides a compound having the structure of formula (II) or (IIA), wherein each of the elements is N, or a pharmaceutically acceptable salt thereof. In one embodiment, X 1 In another embodiment, X 2 In another embodiment, X 3 This is N.

[0180] In one embodiment, the compound has the following structure:

[0181] [ka] (In the formula, n1 is between 0 and 10, and n2 is between 0 and 10.) The present invention provides a compound having the structure of formula (II) or (IIA), represented by , or a pharmaceutically acceptable salt thereof.

[0182] One embodiment is a compound represented by the structure of formula (III):

[0183] [ka] or a pharmaceutically acceptable salt thereof (in the formula, R 1 , R 3 , R 4 , R 5 These are, independently, H, alkyl, alkoxyl, alkenyl, alkynyl, alicyclic, aryl, biaryl, halo, heteroaryl, and -COR. 2x ,

[0184] [ka] And R 2x and R 2y Each of these is independently selected from the group consisting of H, -OH, -CH2-OH, -NH2, -CH2-NH2, -COOMe, -COOH, -CONH2, -COCH3, alkyl, alkenyl, alkynyl, alicyclic, aryl, biaryl, and heteroaryl, R2z Each of these is -NH2, -NR 2q R 2q’ , -OR 2q , -SO-R 2q , and -COR 2q Independently selected from the group consisting of, each R 2q and R 2q’ These are independently alkyl or H,

[0185] [ka] It is a 3-10 member nitrogen-containing non-aromatic monocyclic or bicyclic heterocyclic, and R 21 is H or alkyl, and n' is between 0 and 30; Z is the group of the formula GL-, GL-CO-, GLC(O)-alkyl-, L is the linker, and G is the folate receptor binding ligand; X 1 , X 2 , X 3 Each of them operates independently, CR q or N, and each R q These are independently hydrogen, halogens, or optionally substituted alkyl groups; n is an integer between 0 and 30 (for example, an integer between 0 and 30); m is between 0 and 4. To provide.

[0186] One embodiment is a compound represented by the structure of formula (IIIA):

[0187] [ka] or a pharmaceutically acceptable salt thereof (in the formula, R 1 is optionally substituted alkyl (e.g., acyclic or cyclic) (e.g., optionally substituted with one or more substituents, each substituent independently being a halogen, alkyl, heteroalkyl, alkoxy, or cycloalkyl); Y is H, -OR z , -NR2x R 2y , -SR z -SOR z , -SO3R z -N3, -COR z ,-COOR z ,-CONR z 2. -COSR z , -SO2N(R z )2, or -CON(R z )2, R 2x and R 2y These are, independently, hydrogen and N(R). z )2, -CON(R z )2, -C(R z )2-N(R z )2, -CS-N(R z )2, or optionally substituted alkyl (for example, optionally substituted with one or more substituents, each substituent independently being oxo, halogen, alkyl, heteroalkyl, alkoxy, or cycloalkyl), and each R z is independently hydrogen, halogen, or optionally substituted alkyl; or R 2x and R 2y They combine to form optionally substituted heterocycloalkyls (for example, optionally substituted heterocycloalkyls are monocyclic or bicyclic heterocycloalkyls, and / or optionally substituted heterocycloalkyls are 3- to 10-membered heterocycloalkyls); Each R 3 These are independently halogens, -N3, -CN, -NO2, alkyl, heteroalkyl, cycloalkyl, heterocycloalkyl, alkoxy, aryl, heteroaryl, heterocycloalkyl, amino, hydroxy, carbonyl, or thiol, where alkyl, alkoxy, heteroalkyl, cycloalkyl, or heterocycloalkyl may be substituted; R 4 and R 5Each of these is independently an alkyl, alkoxy, halogen, or cycloalkyl, and the alkyl, alkoxy, and cycloalkyl are optionally substituted; each X 1 , X 2 , and X 3 CR is independent. q or N, and each R q These are independently hydrogen, halogens, or optionally substituted alkyl groups; Z is LG, L is the linker, and G is the folate receptor binding ligand; n is between 1 and 6; m is between 0 and 4. To provide.

[0188] One embodiment provides a compound having the structure of formula (III) or (IIIA), or a pharmaceutically acceptable salt thereof, where n is 1 to 30. In one embodiment, n is 1 to 6. In another embodiment, n is 1 to 3. In another embodiment, n is 1 or 2. In another embodiment, n is 0. In another embodiment, n is 1. In another embodiment, n is 1 and Y is OH. In another embodiment, n is 1 and Y is NH2.

[0189] One embodiment provides a compound having the structure of formula (III) or (IIIA), where Y is OH, or a pharmaceutically acceptable salt thereof.

[0190] One embodiment provides a compound having the structure of formula (III) or (IIIA), where Y is NH2, or a pharmaceutically acceptable salt thereof.

[0191] One embodiment provides a compound having the structure of formula (III) or (IIIA), where n is 1 and Y is OH, or a pharmaceutically acceptable salt thereof.

[0192] One embodiment provides a compound having the structure of formula (III) or (IIIA), where n is 1 and Y is NH2, or a pharmaceutically acceptable salt thereof.

[0193] One embodiment provides a compound having the structure of formula (III) or (IIIA), where n is 0 and Y is NH2, or a pharmaceutically acceptable salt thereof.

[0194] One embodiment is R 1 The present invention provides a compound having the structure of formula (III) or (IIIA), wherein R is optionally substituted with an alkyl group, or a pharmaceutically acceptable salt thereof. In one embodiment, R 1 In some embodiments, R is a substituted C3-C6 alkyl group. 1 R is an acyclic C3-C6 alkyl group which may be substituted. In another embodiment, R 1 It is butyl.

[0195] One embodiment is R 3 The present invention provides a compound having the structure of formula (III) or (IIIA), wherein H is present, or a pharmaceutically acceptable salt thereof.

[0196] One embodiment is R 4 The present invention provides a compound having the structure of formula (III) or (IIIA), wherein R is alkyl, or a pharmaceutically acceptable salt thereof. In one embodiment, R 4 It is methyl.

[0197] One embodiment is R 5 The present invention provides a compound having the structure of formula (III) or (IIIA), wherein R is alkyl, or a pharmaceutically acceptable salt thereof. In one embodiment, R 5 It is methyl.

[0198] One embodiment is R 4 and R 5 The present invention provides a compound having the structure of formula (III) or (IIIA), wherein each of the elements is an alkyl group, or a pharmaceutically acceptable salt thereof. In one embodiment, R 4 and R 5 Each of these is methyl.

[0199] One embodiment provides a compound having the structure of formula (III) or (IIIA), or a pharmaceutically acceptable salt thereof, where m is 0. In another embodiment, m is 1. In another embodiment, m is 2. In another embodiment, m is 3. In another embodiment, m is 4.

[0200] One embodiment is X 1 , X 2 , and X 3 The present invention provides a compound having the structure of formula (III) or (IIIA), wherein each of the elements is N, or a pharmaceutically acceptable salt thereof. In one embodiment, X 1 In another embodiment, X 2 In another embodiment, X 3 This is N.

[0201] In some embodiments, the compound has the structure:

[0202] [ka] It is represented by one or more of the following, or a pharmaceutically acceptable salt thereof.

[0203] In some embodiments, the compound has the structure:

[0204] [ka] It is represented by one or more of the following, or a pharmaceutically acceptable salt thereof.

[0205] In some embodiments, the compound has the following structure:

[0206] [ka] It is represented by or a pharmaceutically acceptable salt thereof.

[0207] Linker The compound may contain an immunomodulator or a pharmaceutically acceptable salt thereof (e.g., a drug) conjugated to the targeting moiety or its radical via a linker (e.g., possibly including a spacer). The linker may be release-type or non-release-type. In some examples, the target of a compound containing a non-release-type linker is an endosome (e.g., of the cell of interest), while the target of a release-type linker is, in some examples, an endosome, cytoplasm, or both (e.g., of the cell of interest).

[0208] In the context of linkers, the term “releasing” means a linker containing at least one bond that can be broken under physiological conditions (e.g., by chemical or enzymatic hydrolysis) by, for example, a reducing agent unstable bond, a pH unstable bond, an acid unstable bond, a base unstable bond, an oxidative unstable bond, a metabolic unstable bond, a biochemical unstable bond, an enzymatic unstable bond, or a p-aminobenzyl polyvalent releasing bond. It is recognized that the physiological conditions resulting in bond breakdown do not necessarily involve biological or metabolic processes, but instead may include standard chemical reactions, such as hydrolysis resulting from compartmentalization into organelles such as endosomes at physiological pH or a pH lower than cytoplasmic pH. A cleavable bond can connect two adjacent atoms in a releasing linker, and / or, for example, at either end or both ends of a releasing linker, other linker or targeting moieties and / or drugs as described herein. In some examples, the releasing linker is broken into two or more fragments. In some examples, the releasing linker is separated from the targeting moiety. In some embodiments, the targeting portion and the immunomodulator are released from each other, and the immunomodulator becomes activated.

[0209] In contrast, the term “non-releasing” in the context of linkers means a linker containing at least one bond that is not easily or rapidly destroyed under physiological conditions. In some embodiments, a non-releasing linker contains a backbone that is stable under physiological conditions (e.g., the backbone is not susceptible to hydrolysis (e.g., aqueous hydrolysis or enzymatic hydrolysis)). In some embodiments, a compound containing a non-releasing linker does not release any component of the compound (e.g., a targeted ligand (e.g., FA-ligand) or an immunomodulator (e.g., a TLR7 agonist)). In some embodiments, a non-releasing linker lacks both disulfide bonds (e.g., SS) and esters in its backbone. In some embodiments, a compound contains a targeted moiety and an immunomodulator linked by a backbone that is substantially stable throughout the entire cycle of the compound (e.g., during endocytosis to target cell endosomes). In some embodiments, compounds containing a non-releasing linker are particularly beneficial when the immunomodulator targets TLRs, nucleotide-binding oligomerization domain (NOD)-like receptors, and / or other pattern recognition receptors present in cellular endosomes. The non-releasing linker may include amides, esters, ethers, amines, and / or thioethers (e.g., thio-maleimide). Specific examples are provided herein, but it will be understood that any molecule(s) can be used as a non-releasing linker, as long as at least one bond is formed that is not easily or rapidly disrupted under physiological conditions.

[0210] In some embodiments, the non-releasing linker includes a linker that hydrolyzes less than 10 percent (e.g., less than 5%, less than 4%, less than 3%, less than 2%, less than 1%, less than 0.1%, less than 0.01%, or less than 0.001%) in an aqueous solution (e.g., buffered (e.g., phosphate buffer)) within a certain period (e.g., 24 hours) at a neutral pH. In some embodiments, when a non-releasing linker is used, less than about 10 percent (10%), preferably less than 5 percent (5%) of the administered conjugate releases free drug, or none at all releases free drug (e.g., into systemic circulation before uptake by targeted cells / tissues). In some embodiments, less than 5 percent (5%) of the free drug is released from the conjugate within 1 hour of administration while the compound is in systemic circulation.

[0211] In some embodiments, the targeting portion does not cleave from the drug / immunomodulator for the compound to become therapeutically effective in vivo. This can be advantageous because, for example, only very small amounts of the drug (e.g., an immunomodulator, e.g., TLR7 or TLR7 / 8 agonist) are released (e.g., systemically) before targeted delivery of the compound, thus enabling the use of targeted compounds and compositions containing potent drugs (e.g., TLR7 and TLR7 / 8 agonists). In some embodiments, tuning the release characteristics of the active ingredient is a difficult aspect of preparing an effective pharmaceutical composition. In some embodiments, compounds containing a non-releasing linker provided herein avoid the difficulty of preparing an effective pharmaceutical composition (e.g., by eliminating the need for release timing). In some embodiments, the immunomodulator / warhead of the compounds provided herein is active when conjugated (e.g., conjugated to a targeted conjugate). In some embodiments, the warhead / immunomodulator is active, while the non-releasing linker and targeting moiety prevent the release (e.g., by the target body) of toxic cytokines (e.g., IL-6) that activate the immune system (e.g., because the compound is specifically targeted (e.g., using folate or an analogue)). In certain specific examples, for example, the warhead / immunomodulator of a compound is active when attached to a non-releasing linker, but the immunomodulator cannot access the appropriate (e.g., targeted) receptor in the cell's endosome until the compound binds to the targeted receptor (e.g., folate receptor).

[0212] Both releasing and non-releasing linkers can be manipulated, for example through PEGylation, according to methodologies commonly known in the art or to be developed in the future, to optimize the biodistribution, bioavailability, and PK / PD (of a compound), and / or increase the uptake (of a compound) into targeted tissue. In some embodiments, the linker is configured to avoid significant release of a pharmaceutically active amount of the drug into circulation before capture by cells (e.g., cells of interest (e.g., macrophages in the fibrous or cancerous tissue being treated)).

[0213] In some embodiments, compounds comprising the release linker of the present disclosure may be designed to diffuse across the endosomal membrane, for example, into the cytoplasm of a targeted cell. In some embodiments, the release linker may be designed so that the immunomodulator is not released until the compound reaches the cytoplasm.

[0214] In some embodiments, the conjugate provided herein includes a release linker (e.g., to facilitate the release of an immunomodulator into the cytoplasm). The release linker can prevent the release of the immunomodulator until, for example, the targeting moiety binds to a suitable target (e.g., a macrophage folate receptor), then translocates to the endosome of the targeting cell and / or diffuses into the cytoplasm (e.g., where the desired pattern recognition receptor is located). In some embodiments, the release linker releases the immunomodulator into the endosome.

[0215] In some embodiments, the linker may include one or more spacers (for example, to facilitate a specific release time, enhance uptake into targeted tissue, and / or optimize the biodistribution, bioavailability, and / or PK / PD of the compound). The spacers may include one or more of alkyl chains, polyethylene glycol (PEG), peptides, sugars, peptidoglycans, clickable linkers (e.g., triazoles), and rigid linkers (e.g., polyproline and polypiperidine).

[0216] In some embodiments, PEG 12 Linkers containing do not completely avoid, but significantly reduce, the nonspecific uptake of the compounds provided herein (e.g., into non-targeted organs (e.g., the liver and / or kidneys of the target after administration)). In some embodiments, the compounds avoid delivery to the liver and kidneys. In some embodiments, the targeted moieties (their free forms, their radicals, or their conjugates) do not bind to uptake receptors on non-targeted cells (e.g., when organs are not target sites, thus stimulation of immune complexes in these organs can be avoided, which is extremely beneficial in clinical situations).

[0217] In some embodiments, a conjugate containing a non-releasing linker reduces or eliminates the toxicity of components released from the conjugate in their free form (e.g., the free form of the compounds and / or ligands provided herein).

[0218] One embodiment provides a compound having the structure of formula (II), (IIA), (IIB), (III), or (IIIA), where L is a cleavable linker, or a pharmaceutically acceptable salt thereof.

[0219] In some embodiments, one or more linkers of the compounds provided herein may include PEG, PEG derivatives, or any other linkers known or to be developed in the art, which can achieve the purposes shown herein. In some embodiments, the linker is repeated n times (where n is a positive integer). For example, n may be any integer selected from the range of 1 to 16, 1 to 32, 1 to 64, or 1 to 96, for example. The number of linker repetitions may be selected to achieve the desired functionality, size, and / or potency of the compound, and / or to take into account the desired application. In some embodiments, one or more linkers include one or more spacers (which may also be used, for example, to specifically design the features of the compound).

[0220] In some embodiments, the linker is a hydrolyzable linker. In some embodiments, the linker is a non-hydrolyzable linker. In some embodiments, the linker is optionally a substituted heteroalkyl. In some embodiments, the linker is a substituted heteroalkyl comprising at least one substituent selected from the group consisting of alkyl, hydroxyl, oxo, PEG, carboxylate, and halo. In some embodiments, the linker comprises a spacer (e.g., as described elsewhere herein).

[0221] One embodiment provides a compound having the structure of formula (II), (IIA), (IIB), (III), or (IIIA), where L is a hydrolyzable linker (e.g., an amide, ester, ether, or sulfonamide), or a pharmaceutically acceptable salt thereof.

[0222] In another embodiment, L is optionally substituted heteroalkyl. In some embodiments, the heteroalkyl is unsubstituted. In other embodiments, the heteroaryl is substituted with at least one substituent selected from the group consisting of alkyl, hydroxyl, acyl, PEG, carboxylate, and halo. In yet another embodiment, L is a substituted heteroalkyl having at least one disulfide bond in its skeleton.

[0223] In another embodiment, L is a peptide or peptidoglycan having at least one disulfide bond in its backbone.

[0224] In another embodiment, L is a cleavable linker that can be cleaved by an enzymatic reaction, reactive oxygen species (ROS), or reducing conditions.

[0225] In some embodiments, L is given by the formula: -NH-CH2-CR 6 R 7 -SS-CH2-CH2-O-CO-(wherein, R 6 and R 7Each of these elements independently comprises H, alkyl, or heteroalkyl.

[0226] In some embodiments, L is given by the formula:

[0227] [ka] (In the formula, p is an integer between 0 and 30; d is an integer between 1 and 40; R 8 and R 9 (Each of these is independently H, alkyl, heterocyclyl, cycloalkyl, aryl, or heteroalkyl.) It is a base of, or contains, this base.

[0228] One embodiment provides a compound having the structure of formula (II), (IIA), (IIB), (III), or (IIIA), where L is a non-releasing linker, or a pharmaceutically acceptable salt thereof.

[0229] One embodiment provides a compound having the structure of formula (II), (IIA), (IIB), (III), or (IIIA), where L is a non-hydrolyzable linker, or a pharmaceutically acceptable salt thereof.

[0230] In some embodiments, L is selected from the group consisting of alkylenes, heteroalkylenes, -O-alkylylenes, alkenylenes, acyls, aryls, heteroaryls, amides, oximes, ethers, esters, triazoles, PEGs, and carboxylates.

[0231] In one embodiment, L is an alkyl ether or contains thereof. In another embodiment, L is an amide or contains thereof. In another embodiment, L is a peptide or peptidoglycan or contains thereof. In another embodiment, L is an amino acid or contains thereof. In another embodiment, L is PEG (e.g., -OCH2-CH2-O-) or contains thereof. In another embodiment, L is a polysaccharide or contains thereof. In another embodiment, L is structure:

[0232] [ka] (In the formula, w is between 0 and 5, and p is between 1 and 30) It is a base represented by, or includes.

[0233] In one embodiment, L is listed below:

[0234] [ka] (In the formula, n'' is between 0 and 30 (for example, n'' is an integer that varies between 0 and 30)) A linker portion selected from, or including, this.

[0235] In some embodiments, the linker is -CONH-CH(COOH)-CH2-SS-CH2-R a R b -O-CO-, -CONH-CH(COOH)R a R b -O-CO-, -C(O)NHCH(COOH)(CH2)2-CONH-CH(COOH)CR a R b -O-CO- or -C(O)NHCH(COOH)(CH2)2-CONH-CH(COOH)-CH2-SS-CH2-R a R b -O-CO-(wherein, R a and R b This independently comprises H, alkyl, or heteroalkyl (e.g., PEG).

[0236] In some embodiments, the linker L is

[0237] [ka] (In the formula, n and m are each independent integers between 0 and 10.) It includes the structure.

[0238] In some embodiments, the linker L is

[0239] [ka] (In the formula, n and m are each independent integers between 0 and 10.) It includes the structure.

[0240] In some embodiments, the linker L is

[0241] [ka] (In the formula, n is an integer between 1 and 32.) It includes the structure.

[0242] In some embodiments, the linker is

[0243] [ka] It includes the structure.

[0244] In some embodiments, the linker is a divalent linker (for example, connecting two groups). In some embodiments, the linker is an ejecting linker. In some embodiments, the linker is a non-ejecting linker.

[0245] The linkers present in the compounds described herein may be any suitable linkers. For example, in some embodiments, the linker is a hydrophilic linker comprising one or more of the following: amino acids (same or different), alkyl chains, PEG monomers, PEG oligomers, PEG polymers, or any combination thereof. In some embodiments, the linker comprises a peptidoglycan, a glycan, or an anionic oligomer.

[0246] For linkers containing one or more PEG units, unless otherwise specified, all carbon and oxygen atoms of the PEG units are part of the backbone. The cleavable bonds of a release linker are part of the backbone. The "backbone" of linker L is G 1 and G 2 It is the shortest chain of continuous atoms that forms a covalent bond between them.

[0247] In some embodiments, the multivalent linker has a branched skeleton, and each branch acts as a section of the skeleton linker until it reaches the end.

[0248] Folate receptor-binding ligands In some cases, the toxicity associated with the systemic administration of at least the conventional drugs identified herein hinders their practical application in the treatment of fibrotic diseases and disorders, cancer, and other disease conditions. For example, TLR agonists may not be tolerable by individuals and, in some cases, may result in death of the subject (e.g., when administered systemically via conventional methods). In some embodiments, compounds, such as those having formulas (I), (IA), (II), (IIA), (IIB), (III), and / or (IIIA), are potent and may be used in conjunction with mechanisms to avoid systemic toxicity (e.g., targeted moieties and / or releasing and non-releasing linkers provided herein).

[0249] In some examples, an immunomodulator or a pharmaceutically acceptable salt thereof is conjugated to the targeting moiety. In some embodiments, the targeting moiety comprises a ligand or other atom or molecule that targets a specific region or tissue of an individual (e.g., with high specificity), and in certain examples, this may include, for example, hormones, antibodies, and / or vitamins. As described in further detail below, in at least one embodiment, the targeting moiety comprises a molecule having (e.g., high) affinity for the folate receptor β (FR-β). In some examples, the targeting moiety may optionally have specific affinity for any receptor specific to cells or tissues of fibrous disease or disorder, or cancer.

[0250] In some cases, FR-β is significantly upregulated in activated myeloid cells (e.g., dominant activated monocytes and M2-like macrophages), and data recorded to date support the idea that FR-β is induced only in myeloid-derived cells after exposure to anti-inflammatory or pro-inflammatory stimuli. Folate receptors can be upregulated in non-mucinous ovarian cancers (e.g., over 90% of them). In certain cases, folate receptors are present in renal cancer, brain cancer, lung cancer, and breast cancer. While some cancers do not express enough folate receptors themselves to provide the desired specificity, the cancerous tumors express myeloid-derived immunosuppressive cells (MDSCs) that express FR-β and can be targeted by the targeting moiety. In some embodiments, folate receptors are substantially absent (e.g., present only at very low levels) in healthy (non-myeloid) tissues (e.g., lungs, liver, spleen, heart, brain, muscle, intestine, pancreas, bladder, etc.). In some cases, uptake of folate-targeted imaging agents occurs in, for example, inflammatory tissue, malignant lesions, and the kidney. In certain cases, non-cancerous subjects simply retain folate-targeted drugs in the kidney and inflammatory sites. In some cases, mismatches in folate receptor expression provide a mechanism for selectively targeting fibrous cancer cells.

[0251] In some embodiments, compounds, compositions, and methods utilize the limited expression of FR-β to target / localize systemically administered potent compounds (e.g., conjugates or drugs) to fibrous and / or cancerous tissues. In some examples, compounds are delivered directly to FR-β expressing cells, for example, to favorably prevent systemic activation of the immune system and to avoid, for example, the toxicity (e.g., at least some of it) that has previously hindered the systemic use of untargeted compounds (e.g., drugs). In some embodiments, for example, the methods are used to treat fibrous diseases and / or cancer, regardless of whether the cancer expresses folate receptors. In some embodiments, folate and other folate receptor-binding ligands (or their radicals), such as folate salts, are used as targeting moieties because they have affinity for FR-β, for example.

[0252] Folic acid is a member of the vitamin B group and can play an essential role in cell survival by participating in the biosynthesis of nucleic acids and amino acids. Folic acid can enhance the specificity of conjugated immunomodulator drugs by targeting activated myeloid cells and conjugated anticancer drugs by targeting folic acid receptor-positive cancer cells. Compounds comprising a folic acid ligand (or its radical), or a functional fragment or analog thereof, and an immunomodulator (e.g., a TLR7 or TLR7 / 8 agonist) as a targeting moiety are provided herein. In some examples, TLR7 and TLR7 / 8 are present in endosomes. In some embodiments, the compound, or its radical, binds to a TLR (e.g., TLR7 or TLR7 / 8).

[0253] Alternatively, pyrido[2,3-d]pyrimidine analog ligands (e.g., or their radicals), their functional fragments or analogs, or any other molecule, fragment, or atom having affinity for FR-β (e.g., high specificity, but not limited to folate) may be used as the targeting moiety (or its radical). For example, such folate analogs may have a relative affinity for bound FR-β of about 0.01 or more compared to folate at about 20°C / 25°C / 30°C / physiological temperatures. Similarly, galectin-3 ligands, translocator protein (TSPO) ligands, and any other ligands or targeting moieties having high specific affinity for fibrous and / or cancerous cells or tissues may be used.

[0254] Specific examples of suitable targeting moieties (or radicals) are provided here; however, it will be understood that the targeting moieties (or radicals) may include any ligand (or radical) useful for targeting FR-β and are not limited to the structures specified herein. The ligand (or radical) may bind to FR-β.

[0255] One embodiment provides a compound having the structure of formula (II), (IIA), (IIB), (III), or (IIIA), in which G is a folate receptor-binding ligand, or a pharmaceutically acceptable salt thereof. In one embodiment, G is a folate, folate, or a functional fragment or derivative thereof, or derived therefrom. In one embodiment, G is a folate or folate derivative. In another embodiment, G is a pteroic acid or pteroyl derivative.

[0256] In some embodiments, G is a reduced folate. In some embodiments, G is a naturally occurring folate. In some embodiments, G is selected from the group consisting of 5-methyltetrahydrofolate (5-MTHF), 5-formyltetrahydrofolate (5-formyl-THF), 10-formyltetrahydrofolate (10-formyl-THF), 5,10-methylenetetrahydrofolate (5,10-methylene-THF), 5,10-methenyltetrahydrofolate (5,10-methenyl-THF), 5,10-formiminotetrahydrofolate (5,10-formimino-THF), 5,6,7,8-tetrahydrofolate (THF), and dihydrofolate (DHF).

[0257] In one embodiment, G is the base of formula (IV):

[0258] [ka] (In the formula, each R is independent,

[0259] [ka] (either it is, or it contains, or R is a naturally occurring or unnatural amino acid or its derivative or fragment.) The present invention provides a compound having the structure of formula (II), (IIA), (IIB), (III), or (IIIA), or a pharmaceutically acceptable salt thereof, which is or contains the same.

[0260] In one embodiment, G has the structure of formula (V):

[0261] [ka] The present invention provides a compound having the structure of formula (II), (IIA), (IIB), (III), or (IIIA), which is a radical having (for example, the group or containing the group), or a pharmaceutically acceptable salt thereof.

[0262] In one embodiment, G has the structure of formula (VI):

[0263] [ka] The present invention provides a compound having the structure of formula (II), (IIA), (IIB), (III), or (IIIA), which is a radical having (for example, the group or containing the group), or a pharmaceutically acceptable salt thereof.

[0264] One embodiment has the following structure:

[0265] [ka]

[0266] [ka]

[0267] [ka] The present invention provides a compound, or a pharmaceutically acceptable salt thereof, represented by one of the following:

[0268] One embodiment has the following structure:

[0269] [ka]

[0270] [ka]

[0271] [ka] The present invention provides a compound, or a pharmaceutically acceptable salt thereof, represented by one of the following:

[0272] One embodiment has the following structure:

[0273] [ka] The present invention provides a compound (compound 5) (e.g., folic acid-PEG3-TLR7-1A containing a release linker) or a pharmaceutically acceptable salt thereof having the above characteristics.

[0274] One embodiment has the following structure:

[0275] [ka] The present invention provides a compound (compound 4) (e.g., folic acid-PEG3-TLR7-1A containing a non-releasing linker) or a pharmaceutically acceptable salt thereof having the above characteristics.

[0276] The compounds can be prepared by conventional methods of organic synthesis performed by those skilled in the art. The general reaction sequences outlined below represent common methods useful for preparing the compounds, but are not intended to limit their scope or usefulness.

[0277] The description of compounds herein is limited by the principles of chemical bonding known to those skilled in the art. Therefore, where a group can be substituted by one or more of several substituents, such substitutions are selected, according to the principles of chemical bonding, to give a compound that is not inherently unstable, and / or would be known to those skilled in the art as being prone to instability under ambient conditions, e.g., aqueous conditions, neutral conditions, and certain known physiological conditions. For example, heterocycloalkyl or heteroaryl groups are bonded to the rest of the molecule via ring heteroatoms according to the principles of chemical bonding known to those skilled in the art, thereby avoiding compounds that are inherently unstable.

[0278] Pharmaceutical composition The compounds described herein may be administered alone or formulated as pharmaceutical compositions comprising one or more compounds with one or more pharmaceutically acceptable excipients. As used herein, the term “composition” generally refers to any product comprising two or more components, including the compounds described herein. It should be understood that the compositions described herein may be prepared from isolated compounds or from salts, solutions, hydrates, solvates, and other forms of the compounds. It is recognized that certain functional groups, such as hydroxy, amino, and similar groups, can form complexes with water and / or various solvents in various physical forms of the compounds. It should also be understood that compositions may be prepared from various amorphous, non-amorphous, partially crystalline, crystalline, and / or other morphological forms of the compounds, and that compositions may be prepared from various hydrates and / or solvates of the compounds. Thus, such pharmaceutical compositions listing compounds may include each of the various morphological forms and / or solvates or hydrate forms of the compounds, or any combination thereof, or their individual forms.

[0279] One embodiment provides a pharmaceutical composition comprising a compound of formula (I), (IA), (II), (IIA), (IIB), (III), or (IIIA), or any compound encompassed by such formulas, or a pharmaceutically acceptable salt thereof, and at least one pharmaceutically acceptable excipient.

[0280] One embodiment provides a pharmaceutical composition comprising an effective amount of a therapeutically (or prophylactically) effective compound of formula (I), (IA), (II), (IIA), (IIB), (III), or (IIIA), or any compound encompassed by such formulas, or a pharmaceutically acceptable salt thereof, and at least one pharmaceutically acceptable excipient.

[0281] The compounds and / or compositions described herein may be administered in unit dosage forms and / or compositions containing one or more pharmaceutically acceptable carriers, adjuvants, diluents, excipients, and / or vehicles, as well as combinations thereof.

[0282] As used herein, the term “administer” generally refers to any means of introducing the compounds described herein into a host subject, including, but not limited to, oral, intravenous, intramuscular, subcutaneous, transdermal, inhalation, buccal, ocular, sublingual, vaginal, rectal, and similar routes of administration.

[0283] The administration of a compound as a salt may be appropriate. Examples of acceptable salts include, but are not limited to, alkali metal (e.g., sodium, potassium, or lithium) or alkaline earth metal (e.g., calcium) salts; however, any salt that is generally non-toxic and effective when administered to the subject being treated is acceptable. Similarly, "pharmaceutically acceptable salts" refer to salts with counterions that can be used in pharmaceuticals. Such salts may include, but are not limited to, (1) acid addition salts obtained by the reaction of a free base of the parent compound with an inorganic acid, such as hydrochloric acid, hydrobromic acid, nitric acid, phosphoric acid, sulfuric acid, perchloric acid, etc., or with an organic acid, such as acetic acid, oxalic acid, (D) or (L) malic acid, maleic acid, methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid, salicylic acid, tartaric acid, citric acid, succinic acid, malonic acid, etc.; or (2) salts formed by either replacing an acidic proton present in the parent compound with a metal ion, such as an alkali metal ion, an alkaline earth ion, or an aluminum ion, or coordinating with an organic base, such as ethanolamine, diethanolamine, triethanolamine, trimethamine, N-methylglucamine, etc. Pharmaceutically acceptable salts are well known to those skilled in the art, and any such pharmaceutically acceptable salt is intended.

[0284] Acceptable salts can be obtained using standard procedures known in the art, which include reacting a sufficiently acidic compound (but not limited to) with a suitable base to obtain a physiologically acceptable anion. Suitable acid addition salts are formed from acids that form non-toxic salts. Examples of non-limiting but representative salts include acetate, aspartate, benzoate, besilate, bicarbonate / carbonate, bisulfate / sulfate, borate, cansilate, citrate, edisylate, esylate, formate, fumarate, gluceptate, gluconate, glucuronate, hexafluorophosphate, hibenzate, hydrochloride / chloride, hydrobromide / bromide, hydroiodide / iodide, isethionate, lactate, malate, maleate, malonate, mesilate, methylsulfate, naphthylate, 2-napsylate, nicotinate, nitrate, orotate, oxalate, palmitate, pamoate, phosphate / hydrogen phosphate / dihydrogen phosphate, saccharate, stearate, succinate, tartrate, tosylate, and trifluoroacetate. Suitable base salts of the compounds described herein are formed from bases that form non-toxic salts. Non-limiting but exemplary examples include salts of arginine, benzathine, calcium, choline, diethylamine, diolamine, glycine, lysine, magnesium, meglumine, olamine, potassium, sodium, tromethamine, and zinc. Hemi salts of acids and bases, such as hemisulfates and hemicalcium salts, may also be formed.

[0285] The compounds may be formulated as pharmaceutical compositions and administered to mammalian hosts, such as human patients, in various forms adapted to a selected route of administration. For example, pharmaceutical compositions may be formulated for oral or parenteral, intravenous, intra-arterial, intraperitoneal, intrathecal, epidural, intraventricular, intraurethral, ​​intrasternal, intracranial, intratumoral, intramuscular, topical, inhalation, and / or subcutaneous routes and administered via these routes. In fact, in at least one embodiment, the compounds and / or compositions may be administered directly into the bloodstream, muscle, or viscera.

[0286] For example, in at least one embodiment, the compound may be administered systemically (e.g., orally) in combination with a pharmaceutically acceptable vehicle, such as an inert diluent or an absorbable edible carrier. For oral therapeutic administration, the active compound may be combined with one or more excipients and used in the form of ingestible tablets, buccal tablets, lozenges, capsules, elixirs, suspensions, syrups, wafers, etc. The percentages of compositions and preparations may vary and be between about 1% and about 99% by weight of the active ingredient(s) and binders, excipients, disintegrants, lubricants, and / or sweeteners (as known in the art). The amount of the active compound in such therapeutically useful compositions is such that an effective dose level is obtained.

[0287] For example, the preparation of parenteral compounds / compositions under sterile conditions by freeze-drying can be easily achieved using standard pharmaceutical techniques well known to those skilled in the art. In at least one embodiment, the solubility of the compound used in the preparation of the parenteral composition can be increased by using appropriate formulation techniques, such as the incorporation of a solubility enhancer.

[0288] As described above, the compounds / compositions can also be administered by injection or by infusion (e.g., using needled (including microneedle) syringes and / or needleless syringes). The solutions of the active compositions are aqueous and may be mixed with non-toxic surfactants, and / or may contain carriers or excipients, such as salts, carbohydrates and buffering agents (preferably pH 3-9), but for some applications they may be better formulated as sterile non-aqueous solutions or in dry form for use with a suitable vehicle, such as sterile, pyrogen-free water or phosphate-buffered saline. For example, dispersions may be prepared in glycerin, liquid PEG, triacetin, and mixtures thereof, as well as in oil. Under normal storage and use conditions, these preparations may further contain preservatives to prevent microbial growth.

[0289] Pharmaceutical dosage forms suitable for injection or infusion may include sterile powders containing active ingredients suitable for immediate preparation of sterile injection or infusion solutions or dispersions, either as sterile aqueous solutions or dispersions, or optionally encapsulated in liposomes. In all cases, the final dosage form should be sterile, fluid, and stable under manufacturing and storage conditions. Liquid carriers or vehicles may be solvents or liquid dispersion media containing, for example, but not limited to, water, ethanol, polyols (e.g., glycerol, propylene glycol, liquid PEG, etc.), vegetable oils, non-toxic glyceryl esters, and / or suitable mixtures thereof. In at least one embodiment, adequate fluidity may be maintained by liposome formation, maintenance of the required particle size in the case of dispersions, or the use of surfactants. Microbial action may be prevented by the addition of various antimicrobial and antifungal agents, such as parabens, chlorobutanol, phenol, sorbic acid, thimerosal, etc. In certain cases, it may be desirable to include one or more isotonic agents, such as sugars, buffers, or sodium chloride. Extended absorption of injectable compositions can be achieved by incorporating drugs formulated to delay absorption, such as aluminum monostearate and gelatin.

[0290] Sterile injectable solutions can be prepared by incorporating the active compound and / or composition into the required amount of a suitable solvent containing, if necessary, one or more of the other components shown above, followed by filtration sterilization. For sterile powders for preparing sterile injectable solutions, preferred preparation methods are vacuum drying and lyophilization, which result in a powder of the active ingredient present in the previously sterile filtered solution + any additional desired ingredients.

[0291] For topical administration, it may be preferable to administer the compound to the skin as a composition or formulation in combination with a dermatologically acceptable carrier, which may be solid or liquid. For example, in certain embodiments, the solid carrier may include finely powdered solids such as talc, clay, microcrystalline cellulose, silica, and alumina. Similarly, a useful liquid carrier may include water, alcohol, glycol, or a water-alcohol / glycol blend, which can dissolve or disperse the compound at an effective level, possibly with the help of a non-toxic surfactant. Furthermore, adjuvants, such as fragrances and antimicrobial agents, may be added to optimize the properties for a given application. The resulting liquid composition may be applied from an absorbent pad, used to impregnate bandages and / or other coverings, sprayed onto a targeted area using a pump or aerosol sprayer, or simply applied directly to the desired area of ​​interest.

[0292] Thickening agents, such as synthetic polymers, fatty acids, fatty acid salts and esters, fatty alcohols, modified cellulose, or modified mineral materials, can also be used with a liquid carrier to form spreadable pastes, gels, ointments, soaps, etc., for direct application to the target skin.

[0293] As used herein, the term “therapeutic (or prophylactic) effective dose” means the amount of a compound administered, either in a single dose or over a course of treatment cycles, that affects the health, well-being, or mortality of a subject (e.g., delaying the onset of one or more signs and / or symptoms associated with fibrous disease or a condition and / or cancer, and / or reducing their severity, unless otherwise specifically stated). Useful doses of the compounds of this disclosure can be determined by comparing their in vitro activity with their in vivo activity in animal models. Methods for extrapolating effective doses in mice and other animals to human subjects are known in the art. In fact, the dose of a compound can vary significantly depending on the host subject’s condition, the cancer or fibrous disease being treated, the stage of the disease, the route and tissue distribution of the compound, and the possibility of concurrent use of other therapeutic measures (e.g., radiotherapy or additional drugs in combination therapy). The amount of a composition required for use in a procedure (e.g., a therapeutic or diagnostically effective dose or size) varies not only by the specific use but also by the selected salt (if applicable) and the characteristics of the subject (e.g., age, condition, sex, body surface area and / or mass of the subject, tolerance to the drug, etc.), and is ultimately at the discretion of the attending physician, clinician, or other person. The therapeutically (or prophylactically) effective or diagnostically effective dose or dosage may range from approximately 0.05 mg / kg of patient weight to approximately 30.0 mg / kg of patient weight, or from approximately 0.01 mg / kg of patient weight to approximately 5.0 mg / kg of patient weight, including, for example, but not limited to, 0.01 mg / kg, 0.02 mg / kg, 0.03 mg / kg, 0.04 mg / kg, 0.05 mg / kg, 0.1 mg / kg, 0.2 mg / kg, 0.3 mg / kg, 0.4 mg / kg, 0.5 mg / kg, 1.0 mg / kg, 1.5 mg / kg, 2.0 mg / kg, 2.5 mg / kg, 3.0 mg / kg, 3.5 mg / kg, 4.0 mg / kg, 4.5 mg / kg, and 5.0 mg / kg (all of which are kg of patient weight). The total therapeutic (or prophylactic) or diagnostically effective dose of the compound may be administered as a single dose or in divided doses, and may, at the physician's discretion, deviate from the typical range given herein.

[0294] In another embodiment, the compound is approximately 0.5 g / m² 2 ~about 500mg / m 2 , about 0.5g / m 2 ~about 300mg / m 2 , or approximately 100g / m 2 ~about 200mg / m 2 It may be administered in a therapeutic (or preventive) or diagnostically effective dose. In other embodiments, the dose is approximately 0.5 mg / m². 2 ~about 500mg / m 2 , about 0.5mg / m 2 ~about 300mg / m 2 , about 0.5mg / m 2 ~about 200mg / m 2 , about 0.5mg / m 2 ~about 100mg / m 2 , about 0.5mg / m 2 ~about 50mg / m 2 , about 0.5mg / m 2 ~about 600mg / m 2 , about 0.5mg / m 2 ~about 6.0mg / m 2 , about 0.5mg / m 2 ~about 4.0mg / m 2 , or approximately 0.5 mg / m² 2 ~about 2.0mg / m 2 This may be the case. The total amount may be administered as a single dose or in divided doses and may, at the physician's discretion, deviate from the typical range given herein. These amounts are based on the area of ​​the body surface m². 2 Based on.

[0295] Treatment method In certain embodiments, the composition and method are useful for the prevention and / or treatment of fibrotic diseases. In certain embodiments, the composition is also useful for the prevention and / or treatment of cancer. The composition and method can utilize strategies to (e.g., selectively) target the innate immune system and reprogram macrophage polarization from M2 to M1, for example, by utilizing its anti-fibrotic properties.

[0296] Vertebrates primarily exhibit two main immune strategies: the innate immune system and the adaptive immune system. The innate, or nonspecific, immune response is the first line of defense against non-self pathogens and consists of physical, chemical, and cellular defenses. The adaptive immune system, on the other hand, acts against pathogens that can evade or overcome the primary innate immune defense.

[0297] Inflammatory responses play a role in immunity. For example, when tissue is damaged or a pathogen is detected, an inflammatory response is initiated and the immune system is mobilized. Immune cells of the innate immune system (e.g., neutrophils and eosinophils) are first mobilized via the vascular and lymphatic systems to the site of tissue injury or damage or to the location of the pathogen, followed by the mobilization of macrophages.

[0298] Cells of the innate immune system can sense specific protein sequences present in microbial pathogens or other non-self cells and express special pattern recognition receptors that bind to them. For example, two classes of molecules that can bind to these pattern recognition receptors are pathogen-associated molecular patterns associated with microbial pathogens and injury-associated molecular patterns associated with host cellular components released during cell damage or cell death. Recognition of these protein sequences by pattern recognition receptors can initiate signaling pathways whose products induce the expression of certain genes whose products control the innate immune response (e.g., ultimately directing the development of antigen-specific adaptive immunity, if necessary). Thus, because pattern recognition receptors mediate these signaling pathways, they can be used to positively or negatively regulate the innate immune response—and even the adaptive immune response.

[0299] More specifically, macrophages are a diverse group of white blood cells known to eliminate pathogens through phagocytosis. Macrophages are broadly classified as having either an M1 or M2 phenotype, depending on which specific differentiation they undergo in response to their local tissue environment.

[0300] In some cases, macrophages are polarized into the M1 phenotype by exposure to IFN-γ, lipopolysaccharide (LPS), and / or granulocyte-macrophage colony-stimulating factor (GM-CSF). In certain cases, the M1 phenotype is characterized by the production of high levels of pro-inflammatory cytokines (e.g., IL-1β, tumor necrosis factor (TNF), IL-12, IL-18, and / or IL-23), the ability to mediate resistance to pathogens, potent microbiota-killing properties, high production of reactive nitrogen and oxygen intermediates, and / or enhancement of the Th1 response. In some cases, M1 polarization is associated with the “attack and killing” phase of the innate immune response. In certain cases, M1 polarization functions to inhibit or prevent the initial establishment of infection and / or to remove damaged tissue.

[0301] In certain cases, after the innate immune system has carried out this “attack and kill” phase, macrophages can reprogram themselves to become part of the healing system. In some cases, macrophages release growth factors to promote healing. Such growth factors may include (but are not limited to) certain cytokines, such as IL-4, IL-10, platelet-derived growth factor (PDGF), TGFβ, CCL18, and / or IL-13. In certain cases, exposure to such cytokines / growth factors may activate the M2 macrophage phenotype.

[0302] In contrast to M1 macrophages, M2 macrophages are typically associated with wound healing and tissue repair. In some cases, M2 macrophages are characterized by their involvement in tissue remodeling, immunomodulation / suppression, and / or tumor promotion. Specifically, M2 macrophages produce polyamines that induce cell proliferation and / or proline that induces collagen production. While this healing response is beneficial in healthy subjects, the presence of M2 macrophages can have significantly detrimental effects in subjects with fibrous disease or cancer, through immunosuppression and / or promotion of tumor growth and fibrosis.

[0303] For example, fibrotic pathology may begin with an unknown trauma or injury to the epithelium. Depending on the resulting tissue damage, chemokines and other factors may be released, for example, exhibiting an M2-like phenotype and promoting the infiltration of immune cells, including monocytes and macrophages that release pro-fibrotic cytokines (e.g., innate immune response). Chronic secretion of these cytokines may then activate resident and invasive fibroblasts / fibrocytes that become myofibroblasts, which in turn secrete collagen and other extracellular matrix proteins that can harden the surrounding tissue. In this way, these M2 macrophages can exacerbate the disease and act pro-fibrotic. For example, M2 macrophages may infiltrate the lungs of patients with idiopathic pulmonary fibrosis (IPF), for instance, and promote fibrosis within them, accelerating disease progression.

[0304] Growth factors and other cytokines produced by the M2 phenotype can drive cancerous tumor growth through similar pathways. For example, cancer may also be accompanied by an anti-inflammatory immune response that promotes cancerous tumor growth (e.g., by growth factors secreted by activated M2-like macrophages, specifically tumor-associated macrophages (TAMs)) and / or promotes collagen formation in cancerous tumors (e.g., through downstream fibrous collagen production). In certain cases, this can lead to more advanced and more difficult-to-treat cancerous tumors, as their growth reduces their drug permeability.

[0305] Reprogramming of M2-like macrophages into M1-like macrophages In certain cancers and fibrous diseases, macrophages exhibit a disproportionate bias toward an anti-inflammatory pro-fibrotic (M2-like) phenotype. In certain cases, immunomodulators (e.g., TLR7 agonists) can convert—e.g., reprogram—activated myeloid cells (e.g., M2-like macrophages) toward pro-inflammatory and anti-fibrotic M1 polarization (e.g., where these cells produce little to no growth factors and / or related cytokines, e.g., delaying or even eliminating the progression of the disease state). Compounds, compositions, and methods for reversing the shift from anti-fibrotic / pro-inflammatory macrophages to pro-fibrotic / anti-inflammatory (M1>M2) macrophages observed during the development of fibrous diseases (e.g., IPF) and certain cancers are provided herein. In some embodiments, the compounds, compositions, and methods described herein reduce the levels of pro-fibrotic / anti-inflammatory biomarkers (e.g., pro-fibrotic activity (e.g., CCL18, hydroxyproline, and collagen)) in an individual or a sample thereof. In some embodiments, the compounds, compositions, and methods described herein increase anti-fibrotic / pro-inflammatory biomarkers (e.g., TNFα and IFN-γ). In some embodiments, the compounds, compositions, and methods described herein reprogram M2-like macrophages into M1-like macrophages. In some embodiments, the compounds, compositions, and methods described herein alter cytokine secretion and chemotactic factors produced by macrophages. In some embodiments, compositions are provided that reverse the M2-like phenotype shift to provide effective treatments for fibrotic diseases, disorders, or conditions thereof, as well as certain types of cancer.

[0306] In at least one embodiment, a drug comprising an immunomodulator (e.g., a TLR7 agonist) is used to prepare the compounds and compositions used in the present method. A therapeutic agent (e.g., a drug) suitable for reprogramming activated macrophages (M2-like phenotype) into an M1-like phenotype (e.g., a TLR7 agonist) can be used, and the drug (or warhead) can function in the endosomes and / or cytoplasm of cells (e.g., depending on its structure). In at least one embodiment, the therapeutic agent (e.g., a drug) is an immunomodulator (e.g., one that positively regulates pattern recognition receptors and / or their downstream signaling pathways (in each case, part of the innate immune system)), e.g., a TLR7 agonist. In some embodiments, the drug is a TLR7 agonist according to any one of formulas (I), (IA), (II), (IIA), (IIB), (III), or (IIIA).

[0307] In certain embodiments, a compound is provided comprising a targeting moiety (or radical thereof; e.g., folic acid) bound to an immunomodulator (or radical thereof; e.g., a TLR7 agonist) that targets a cellular pattern recognition receptor, wherein the targeting moiety comprises a folic acid ligand or a functional fragment or analog thereof. In some embodiments, the immunomodulator is a TLR7 agonist.

[0308] One embodiment provides a method for treating cancer in an individual in need. The method comprises administering to an individual in need one or more compounds of any one of the formulas (I), (IA), (II), (IIA), (IIB), (III), or (IIIA), any compound encompassed by such formulas, or a composition containing the same, in a therapeutically effective amount. In some embodiments, the cancer is characterized by a tumor containing TAM. In some embodiments, cancer can be lung cancer, bone cancer, pancreatic cancer, skin cancer, head cancer, cervical cancer, cutaneous melanoma, intraocular melanoma, uterine cancer, ovarian cancer, endometrial cancer, epithelial cancer, leiomyosarcoma, rectal cancer, stomach cancer, colon cancer, breast cancer, triple-negative breast cancer, fallopian tube cancer, endometrial cancer, cervical cancer, vaginal cancer, vulvar cancer, Hodgkin's disease, esophageal cancer, small intestine cancer, endocrine cancer, thyroid cancer, parathyroid cancer, non-small cell lung cancer, small cell lung cancer, adrenal cancer, soft tissue sarcoma, urethral cancer, penile cancer, prostate cancer, chronic leukemia, acute leukemia, lymphocytic lymphoma, pleural mesothelioma, bladder cancer, gastric cancer. The cancer is selected from the group consisting of Burkitt lymphoma, ureteral cancer, kidney cancer, renal cell carcinoma, renal pelvis cancer, central nervous system (CNS) neoplasms, primary CNS lymphoma, vertebral axial tumor, brainstem glioma, pituitary adenoma, cholangiocarcinoma, Haasle cell thyroid cancer, and gastroesophageal junction adenocarcinoma. In certain embodiments, the cancer is lung cancer, breast cancer, colon cancer, ovarian cancer, pancreatic cancer, or epithelial cancer.

[0309] In one embodiment, the cancer is lung cancer.

[0310] In one embodiment, the cancer is lung cancer, triple-negative breast cancer, colon cancer, stomach cancer, bladder cancer, prostate cancer, or pancreatic cancer.

[0311] One embodiment provides a method for treating an inflammatory disease or disorder. The method includes administering to a patient in need one or more compounds of any one of the formulas (I), (IA), (II), (IIA), (IIB), (III), or (IIIA), any compound encompassed by such formulas, or a composition containing the same.

[0312] One embodiment provides a method for treating a fibrous disease or disorder in an individual in need. The method includes administering to an individual in need one or more compounds of any one of the formulas (I), (IA), (II), (IIA), (IIB), (III), or (IIIA), any compound encompassed by such formulas, or a composition containing the same, in a therapeutically effective amount.

[0313] In certain embodiments, the fibrous disease or disorder is selected from the group consisting of articular fibrosis, autoimmune pancreatitis, bladder fibrosis, chronic kidney disease, chronic wound, Crohn's disease, tendonoid, Dupuytren's contracture, endometrial fibroma, fibromatosis, graft-versus-host disease (GVHD), cardiac fibrosis, keloids, hepatic fibrosis (e.g., non-alcoholic steatohepatitis (NASH) or cirrhosis), mediastinal fibrosis, myelofibrosis, nephrogenic systemic fibrosis, Peyronie's disease, pulmonary fibrosis, retroperitoneal fibrosis, scleroderma or systemic sclerosis, and cutaneous fibrosis.

[0314] In certain embodiments, the fibrotic disease or disorder is idiopathic pulmonary fibrosis (IPF), hepatic fibrosis, myelofibrosis, or cardiac fibrosis.

[0315] In other embodiments, the fibrous disease or disorder or inflammatory disease or disorder is selected from the group consisting of lupus, inflammatory bowel disease (IBS), Addison's disease, Graves' disease, Sjögren's syndrome, celiac disease, Hashimoto's thyroiditis, myasthenia gravis, autoimmune vasculitis, reactive arthritis, psoriatic arthritis, pernicious anemia, ulcerative colitis, rheumatoid arthritis, type 1 diabetes mellitus, multiple sclerosis, transplant rejection, fatty liver disease, asthma, osteoporosis, sarcoidosis, ischemia-reperfusion injury, prosthetic osteolysis, glomerulonephritis, scleroderma, psoriasis, autoimmune myocarditis, spinal cord injury, central nervous system disorders, viral infections, influenza, coronavirus infection, cytokine storm syndrome, bone injury, inflammatory brain disease, and atherosclerosis.

[0316] One embodiment provides a method for inhibiting or reducing an inflammatory disease or disorder. The method comprises the step of administering a compound of formula (I), (IA), (II), (IIA), (IIB), (III), or (IIIA), any compound encompassed by such formulas, or a composition containing the same, to a patient in need.

[0317] One embodiment provides a compound of formula (I), (IA), (II), (IIA), (IIB), (III), or (IIIA), any compound encompassed by such formulas, or a composition containing such a compound, for use in a method of treating cancer (for example, any of the types of cancer listed herein).

[0318] One embodiment provides a compound of formula (I), (IA), (II), (IIA), (IIB), (III), or (IIIA), any compound encompassed by such formulas, or a composition containing such compounds, for use in a method of treating an inflammatory disease or disorder. In some embodiments, the inflammatory disease or disorder is a fibrous disease or disorder. In certain embodiments, the fibrous disease or disorder is IPF, hepatic fibrosis, myelofibrosis, or cardiac fibrosis.

[0319] One embodiment provides a compound of formula (I), (IA), (II), (IIA), (IIB), (III), or (IIIA), any compound encompassed by such formulas, or a composition containing such a compound, for use in a method of inhibiting or reducing cancer.

[0320] One embodiment provides a compound of formula (I), (IA), (II), (IIA), (IIB), (III), or (IIIA), any compound encompassed by such formulas, or a composition containing such a compound, for use in a method of inhibiting or reducing fibrosis.

[0321] One embodiment provides the use of compounds of formula (I), (IA), (II), (IIA), (IIB), (III), or (IIIA), any compound encompassed by such formulas, or compositions containing such compounds, for manufacturing pharmaceuticals for treating cancer.

[0322] One embodiment provides the use of compounds of formula (I), (IA), (II), (IIA), (IIB), (III), or (IIIA), any compounds encompassed by such formulas, or compositions containing such compounds, for use in a method of manufacturing pharmaceuticals for treating inflammatory diseases. In some embodiments, the inflammatory disease or disorder is a fibrous disease or disorder.

[0323] A method is provided for inhibiting or reducing fibrosis (for example, in an individual in need, e.g., an individual suffering from cancer or a fibrotic disease), comprising the step of administering to an individual in need (e.g., to the individual) one or more compounds of formula (I), (IA), (II), (IIA), (IIB), (III), or (IIIA), or any compound encompassed by such formulas, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition containing the same, in an amount effective to convert a population of macrophages biased toward an M2-like phenotype (e.g., pro-fibrotic / anti-inflammatory) to an M1-like phenotype (e.g., anti-fibrotic / pro-inflammatory), wherein the population of macrophages is located at a targeted site within the individual, the M2-like phenotype is associated with an anti-inflammatory / pro-fibrotic state, and the M1-like phenotype is associated with an anti-inflammatory / anti-fibrotic state.

[0324] Furthermore, a method is provided for inhibiting or reducing cancerous growth (for example, in an individual in need, e.g., an individual with cancer), comprising the step of administering to an individual in need (e.g., to the individual) an amount effective in converting a population of macrophages biased toward an M2-like phenotype (e.g., pro-fibrotic / anti-inflammatory) to an M1-like phenotype (e.g., anti-fibrotic / pro-inflammatory), wherein the population of macrophages is located at a target site within the individual, the M2-like phenotype is associated with an anti-inflammatory / pro-fibrotic state, and the M1-like phenotype is associated with an anti-inflammatory / anti-fibrotic state. In at least one embodiment, the target site is the tumor microenvironment.

[0325] One embodiment provides the use of compounds of formula (I), (IA), (II), (IIA), (IIB), (III), or (IIIA), any compound encompassed by such formulas, or compositions containing such compounds, for manufacturing pharmaceuticals for treating cancer.

[0326] In certain embodiments, a method for treating lung cancer in an individual requiring such treatment is provided herein, comprising the step of administering to the individual a therapeutically effective amount of a compound of formula (I), (IA), (II), (IIA), (IIB), (III), or (IIIA), any compound encompassed by such formula, or a composition containing such compound, thereby treating the lung cancer. In some embodiments, the lung cancer comprises a tumor containing M2-type (e.g., M2-like phenotype) TAM. In some embodiments, M2 macrophages are reprogrammed to an M1 phenotype by administering a compound of formula (I), (IA), (II), (IIA), (IIB), (III), or (IIIA), any compound encompassed by such formula, or a composition containing such compound.

[0327] In certain embodiments, a method for treating triple-negative breast cancer in an individual requiring such treatment is provided herein, comprising the step of administering to the individual a therapeutically effective amount of a compound of formula (I), (IA), (II), (IIA), (IIB), (III), or (IIIA), any compound encompassed by such formula, or a composition containing such compound, thereby treating the triple-negative breast cancer. In some embodiments, the triple-negative breast cancer comprises a tumor containing M2-type TAM. In some embodiments, M2 macrophages are reprogrammed to M1 type by administering a compound of formula (I), (IA), (II), (IIA), (IIB), (III), or (IIIA), any compound encompassed by such formula, or a composition containing such compound.

[0328] In certain embodiments, a method for treating colon cancer in an individual requiring such treatment is provided herein, comprising the step of administering to the individual a therapeutically effective amount of a compound of formula (I), (IA), (II), (IIA), (IIB), (III), or (IIIA), any compound encompassed by such formula, or a composition containing such compound, thereby treating the colon cancer. In some embodiments, the colon cancer comprises a tumor containing M2 type TAM. In some embodiments, M2 macrophages are reprogrammed to M1 type by administering a compound of formula (I), (IA), (II), (IIA), (IIB), (III), or (IIIA), any compound encompassed by such formula, or a composition containing such compound.

[0329] In certain embodiments, a method for treating gastric cancer in an individual requiring such treatment is provided herein, comprising the step of administering to the individual a therapeutically effective amount of a compound of formula (I), (IA), (II), (IIA), (IIB), (III), or (IIIA), any compound encompassed by such formula, or a composition containing such compound, thereby treating the gastric cancer. In some embodiments, the gastric cancer comprises a tumor containing M2 type TAM. In some embodiments, M2 macrophages are reprogrammed to M1 type by administering a compound of formula (I), (IA), (II), (IIA), (IIB), (III), or (IIIA), any compound encompassed by such formula, or a composition containing such compound.

[0330] In certain embodiments, a method for treating prostate cancer in an individual requiring such treatment is provided herein, comprising the step of administering to the individual a therapeutically effective amount of a compound of formula (I), (IA), (II), (IIA), (IIB), (III), or (IIIA), any compound encompassed by such formula, or a composition containing such compound, thereby treating the prostate cancer. In some embodiments, the prostate cancer comprises a tumor containing M2 type TAM. In some embodiments, M2 macrophages are reprogrammed to M1 type by administering a compound of formula (I), (IA), (II), (IIA), (IIB), (III), or (IIIA), any compound encompassed by such formula, or a composition containing such compound.

[0331] In certain embodiments, a method for treating bladder cancer in an individual requiring such treatment is provided herein, comprising the step of administering to the individual a therapeutically effective amount of a compound of formula (I), (IA), (II), (IIA), (IIB), (III), or (IIIA), any compound encompassed by such formula, or a composition containing such compound, thereby treating the bladder cancer. In some embodiments, the bladder cancer comprises a tumor containing M2 type TAM. In some embodiments, M2 macrophages are reprogrammed to M1 type by administering a compound of formula (I), (IA), (II), (IIA), (IIB), (III), or (IIIA), any compound encompassed by such formula, or a composition containing such compound.

[0332] In certain embodiments, a method for treating pancreatic cancer in an individual requiring such treatment is provided herein, comprising the step of administering to the individual a therapeutically effective amount of a compound of formula (I), (IA), (II), (IIA), (IIB), (III), or (IIIA), any compound encompassed by such formula, or a composition containing such compound, thereby treating the pancreatic cancer. In some embodiments, the pancreatic cancer comprises a tumor containing M2 type TAM. In some embodiments, M2 macrophages are reprogrammed to M1 type by administering a compound of formula (I), (IA), (II), (IIA), (IIB), (III), or (IIIA), any compound encompassed by such formula, or a composition containing such compound.

[0333] One embodiment provides the use of compounds of formula (I), (IA), (II), (IIA), (IIB), (III), or (IIIA), any compound encompassed by such formulas, or compositions containing such compounds, for the manufacture of pharmaceuticals for inhibiting or reducing fibrosis.

[0334] In one embodiment, fibrosis is pulmonary fibrosis (e.g., IPF), hepatic fibrosis, or cardiac fibrosis. In some embodiments, fibrosis is selected from the group consisting of fatty liver disease, cirrhosis, colitis, chronic liver disease, cardiac fibrosis, and scleroderma. In certain embodiments, the fibrotic disease or disorder is pulmonary fibrosis, hepatic fibrosis, scleroderma, myelofibrosis, Crohn's disease, or chronic kidney disease.

[0335] In certain embodiments, a method for treating pulmonary fibrosis in an individual requiring such treatment is provided herein, comprising the step of administering to the individual a therapeutically effective amount of a compound of formula (I), (IA), (II), (IIA), (IIB), (III), or (IIIA), any compound encompassed by such formula, or a composition containing such compound, thereby treating the pulmonary fibrosis. In some embodiments, the pulmonary fibrosis comprises M2-type pro-fibrotic macrophages. In some embodiments, the M2 macrophages are reprogrammed to M1 type by administering a compound of formula (I), (IA), (II), (IIA), (IIB), (III), or (IIIA), any compound encompassed by such formula, or a composition containing such compound.

[0336] In certain embodiments, a method for treating hepatic fibrosis in an individual in need is provided herein, comprising the step of administering to the individual a therapeutically effective amount of a compound of formula (I), (IA), (II), (IIA), (IIB), (III), or (IIIA), any compound encompassed by such formula, or a composition containing such compound, thereby treating the hepatic fibrosis. In some embodiments, the hepatic fibrosis comprises M2-type pro-fibrotic macrophages. In some embodiments, the M2 macrophages are reprogrammed to M1 type by administering a compound of formula (I), (IA), (II), (IIA), (IIB), (III), or (IIIA), any compound encompassed by such formula, or a composition containing such compound.

[0337] In certain embodiments, a method for treating scleroderma in an individual requiring such treatment is provided herein, comprising the step of administering to the individual a therapeutically effective amount of a compound of formula (I), (IA), (II), (IIA), (IIB), (III), or (IIIA), any compound encompassed by such formula, or a composition containing such compound, thereby treating the scleroderma. In some embodiments, the scleroderma comprises M2-type pro-fibrotic macrophages. In some embodiments, the M2 macrophages are reprogrammed to M1 type by administering a compound of formula (I), (IA), (II), (IIA), (IIB), (III), or (IIIA), any compound encompassed by such formula, or a composition containing such compound.

[0338] In certain embodiments, a method for treating myelofibrosis in an individual requiring such treatment is provided herein, comprising the step of administering to the individual a therapeutically effective amount of a compound of formula (I), (IA), (II), (IIA), (IIB), (III), or (IIIA), any compound encompassed by such formula, or a composition containing such compound, thereby treating the myelofibrosis. In some embodiments, the myelofibrosis comprises M2-type pro-fibrotic macrophages. In some embodiments, the M2 macrophages are reprogrammed to M1 type by administering a compound of formula (I), (IA), (II), (IIA), (IIB), (III), or (IIIA), any compound encompassed by such formula, or a composition containing such compound.

[0339] In certain embodiments, a method for treating Crohn's disease in an individual in need is provided herein, comprising the step of administering to the individual a therapeutically effective amount of a compound of formula (I), (IA), (II), (IIA), (IIB), (III), or (IIIA), any compound encompassed by such formula, or a composition containing such compound, thereby treating Crohn's disease. In some embodiments, Crohn's disease comprises M2-type pro-fibrotic macrophages. In some embodiments, the M2 macrophages are reprogrammed to M1 type by administering a compound of formula (I), (IA), (II), (IIA), (IIB), (III), or (IIIA), any compound encompassed by such formula, or a composition containing such compound.

[0340] In certain embodiments, a method for treating chronic kidney disease in an individual requiring it is provided herein, comprising the step of administering to the individual a therapeutically effective amount of a compound of formula (I), (IA), (II), (IIA), (IIB), (III), or (IIIA), any compound encompassed by such formula, or a composition containing such compound, thereby treating the chronic kidney disease. In some embodiments, the chronic kidney disease comprises M2-type pro-fibrotic macrophages. In some embodiments, the M2 macrophages are reprogrammed to M1 type by administering a compound of formula (I), (IA), (II), (IIA), (IIB), (III), or (IIIA), any compound encompassed by such formula, or a composition containing such compound.

[0341] In some embodiments of any method for treating cancer or fibrous disease or disorder disclosed herein, the method (e.g., administration of one or more compounds) does not induce unwanted inflammation in the individual.

[0342] In some embodiments of any method for treating cancer or fibrous disease or disorder disclosed herein, the method further includes the step of administering a second therapeutic agent. In some embodiments, the second therapeutic agent is an anti-inflammatory agent. In some embodiments, the second therapeutic agent is a pro-inflammatory agent (for example, if the method is for treating fibrous disease or disorder). In some embodiments, the second therapeutic agent is a chemotherapeutic agent (for example, if the method is for treating cancer). In some embodiments, the compounds or compositions of the disclosure are administered simultaneously or sequentially in combination with the second therapeutic agent.

[0343] All patents, patent application publications, journal articles, textbooks, and other publications referenced herein represent the level of skill of a person skilled in the art to which this disclosure pertains. All such publications are incorporated herein by reference to the same extent as each individual publication is specifically and individually indicated to be incorporated by reference.

[0344] While certain embodiments of this disclosure are shown and described herein, it will be apparent to those skilled in the art that such embodiments are provided merely as examples. It is intended that the claimed invention is not limited by the specific examples provided herein.

[0345] While the present invention is described with reference to the above specification, the descriptions and examples of embodiments herein are not intended to be constrained. Numerous variations, modifications, and substitutions will be conceivable to those skilled in the art without departing from the present invention. Furthermore, it will be understood that all aspects of the present invention are not limited to the specific descriptions, configurations, or relative proportions shown herein and depend on various conditions and variables. It should be understood that various alternatives to the embodiments of the present invention described herein may be used in the practice of the present invention. Thus, the present invention is intended to encompass any such alternatives, modifications, variations, or equivalents. The following claims define the scope of the present invention, and methods and structures within the scope of these claims, as well as their equivalents, are intended to be covered thereby.

[0346] A certain definition Where used herein and in the appended claims, the singular forms “a,” “and,” and “the” include multiple references unless explicitly indicated in the context. Thus, for example, a reference to “a compound” includes multiple such compounds. Where a range is used herein for a physical property, e.g., molecular weight, or a chemical property, e.g., chemical formula, it is intended to include all combinations and partial combinations of the range, as well as specific embodiments therein. Where the term “about” refers to a number or numerical range, it means that the number or numerical range referred to is an approximation within experimental variability (or statistical experimental error), and therefore the number or numerical range may vary between 1% and 15% of the specified number or numerical range. The term “comprising” (and related terms, e.g., “comprise” or “comprises” or “having” or “including”) is not intended to exclude any embodiments of any compound, composition, method, process, etc. that “consist of” or “essentially consist of” the described features.

[0347] The terms “to treat,” “to treat,” or “treatment” include reducing, alleviating, mitigating, improving, relieving, or shrinking symptoms associated with cancer, fibrous disease or disorder, or an inflammatory condition or disease, in either a chronic or acute treatment scenario. In some embodiments, treatment of fibrous disease or disorder includes reducing fibrosis. In some embodiments, treatment of cancer includes reducing the number of M2-like macrophages found in the associated tumor.

[0348] As used herein, the terms “patient,” “subject,” and “individual” are interchangeable. None of these terms require the supervision of a healthcare professional. For example, administration to an individual includes the individual administering a therapeutic agent to itself, as well as a healthcare professional administering a therapeutic agent to an individual.

[0349] "Alicyclic" refers to a radical that has at least one whole carbon ring, which may be complete or partially saturated, and which may optionally have one or more linear groups attached to it. For example, cycloalkyl and cycloalkenyl groups, such as cyclobutyl and cyclohexa-3-enyl, are considered alicyclic, as are cycloalkyl groups to which one or more linear alkyl groups are attached, such as cyclohexylmethyl, 3-n-propylcyclopenta-2-enylmethyl, or 2,3,4-trimethylcyclohexyl. The bond site of an alicyclic radical may be on a ring atom or on a carbon atom.

[0350] "Alkyl" consists only of carbon and hydrogen atoms, does not contain unsaturated atoms, and has 1 to 15 carbon atoms (for example, C1 to C15). 15 A alkyl group refers to a linear, branched, or cyclic hydrocarbon chain radical. In various embodiments, alkyl groups consist of 3 to 6 carbon atoms (e.g., C3-C6 alkyl groups), 1 to 13 carbon atoms (e.g., C1-C6 alkyl groups), and 13Alkyl), 1-8 carbon atoms (e.g., C1-C8 alkyl), 1-5 carbon atoms (e.g., C1-C5 alkyl), 1-4 carbon atoms (e.g., C1-C4 alkyl), 1-3 carbon atoms (e.g., C1-C3 alkyl), 1-2 carbon atoms (e.g., C1-C2 alkyl), 1 carbon atom (e.g., C1 alkyl), 5-15 carbon atoms (e.g., C5-C 15 The alkyl group comprises 5-8 carbon atoms (e.g., C5-C8 alkyl), 2-5 carbon atoms (e.g., C2-C5 alkyl), or 3-5 carbon atoms (e.g., C3-C5 alkyl). In other embodiments, the alkyl group is selected from methyl, ethyl, 1-propyl (n-propyl), 1-methylethyl (iso-propyl), 1-butyl (n-butyl), 1-methylpropyl (sec-butyl), 2-methylpropyl (iso-butyl), 1,1-dimethylethyl (tert-butyl), and 1-pentyl (n-pentyl). The alkyl group is bonded to the rest of the molecule by a single bond. Unless otherwise specifically stated herein, the alkyl group is optionally substituted by one or more of the following substituents: halo, cyano, nitro, oxo, thioxo, imino, oxymo, trimethylsilanyl, -OR a , -SR a -OC(O)-R a , -N(R a )2, -C(O)R a , -C(O)OR a ,-C(O)N(R a )2, -N(R a )C(O)OR a -OC(O)-N(R a )2, -N(R a )C(O)R a , -N(R a )S(O) t R a (wherein t is 1 or 2), -S(O) t Ure a (wherein t is 1 or 2), -S(O) t R a (wherein the formula t is 1 or 2) and -S(O) t N(R a)2 (wherein t is 1 or 2) (wherein each R a These are independently hydrogen, alkyl (optionally substituted with halogen, hydroxy, methoxy, or trifluoromethyl), fluoroalkyl, carbocyryl (optionally substituted with halogen, hydroxy, methoxy, or trifluoromethyl), carbocyrylalkyl (optionally substituted with halogen, hydroxy, methoxy, or trifluoromethyl), aryl (optionally substituted with halogen, hydroxy, methoxy, or trifluoromethyl), aralkyl (optionally substituted with halogen, hydroxy, methoxy, or trifluoromethyl), heterocyclyl (optionally substituted with halogen, hydroxy, methoxy, or trifluoromethyl), heterocyclylalkyl (optionally substituted with halogen, hydroxy, methoxy, or trifluoromethyl), heteroaryl (optionally substituted with halogen, hydroxy, methoxy, or trifluoromethyl), or heteroarylalkyl (optionally substituted with halogen, hydroxy, methoxy, or trifluoromethyl).

[0351] Alkyl groups can include carbocyclic or carbocyclyl groups. “Carbocyclyl” refers to a stable, non-aromatic monocyclic or polycyclic hydrocarbon radical consisting only of carbon and hydrogen atoms, including condensed or bridging ring systems having 3 to 15 carbon atoms. In certain embodiments, carbocyclyl contains 3 to 10 carbon atoms. In other embodiments, carbocyclyl contains 5 to 7 carbon atoms. Carbocyclyl is bonded to the rest of the molecule by single bonds. Carbocyclyl can be saturated (i.e., containing only single C-C bonds) or unsaturated (i.e., containing one or more double or triple bonds). Fully saturated carbocyclyl radicals are also called “cycloalkyl.” Examples of monocyclic cycloalkyls include, for example, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, and cyclooctyl. Unsaturated carbocyclyls are also called “cycloalkenyl.” Examples of monocyclic cycloalkenyls include, for example, cyclopentenyl, cyclohexenyl, cycloheptenyl, and cyclooctenyl. Polycyclic carbocyclyl radicals include, for example, adamantyl, norbornyl (i.e., bicyclo[2.2.1]heptanyl), norborneyl, dekalinyl, 7,7-dimethyl-bicyclo[2.2.1]heptanyl, etc. Unless otherwise specifically stated herein, the term "carbocyclyl" includes alkyl, alkenyl, alkynyl, halo, fluoroalkyl, oxo, thioxo, cyano, nitro, optionally substituted aryl, optionally substituted aralkyl, optionally substituted aralkenyl, optionally substituted aralquinyl, optionally substituted carbocyclyl, optionally substituted carbocyclylalkyl, optionally substituted heterocyclyl, optionally substituted heterocyclylalkyl, optionally substituted heteroaryl, optionally substituted heteroarylalkyl, -R b -OR a ,-R b -OC(O)-R a ,-R b -OC(O)-OR a ,-R b -OC(O)-N(Ra )2, -R b -N(R a )2, -R b -C(O)R a ,-R b -C(O)OR a ,-R b -C(O)N(R a )2, -R b -OR c -C(O)N(R a )2, -R b -N(R a )C(O)OR a ,-R b -N(R a )C(O)R a ,-R b -N(R a )S(O) t R a (wherein the formula t is 1 or 2), -R b -S(O) t R a (wherein the formula t is 1 or 2), -R b -S(O) t Ure a (wherein the formula t is 1 or 2) and -R b -S(O) t N(R a )2 (wherein t is 1 or 2) (wherein each R aR is independently hydrogen, alkyl (optionally substituted with halogen, hydroxy, methoxy, or trifluoromethyl), fluoroalkyl, cycloalkyl (optionally substituted with halogen, hydroxy, methoxy, or trifluoromethyl), cycloalkylalkyl (optionally substituted with halogen, hydroxy, methoxy, or trifluoromethyl), aryl (optionally substituted with halogen, hydroxy, methoxy, or trifluoromethyl), aralkyl (optionally substituted with halogen, hydroxy, methoxy, or trifluoromethyl), heterocyclyl (optionally substituted with halogen, hydroxy, methoxy, or trifluoromethyl), heterocyclylalkyl (optionally substituted with halogen, hydroxy, methoxy, or trifluoromethyl), heteroaryl (optionally substituted with halogen, hydroxy, methoxy, or trifluoromethyl), or heteroarylalkyl (optionally substituted with halogen, hydroxy, methoxy, or trifluoromethyl), and each R b R is independently, directly bonded, or a linear or branched alkylene or alkenylene chain. c The material is intended to comprise a carbocyclyl radical (which is a linear or branched alkylene or alkenylene chain) optionally substituted by one or more substituents independently selected from the linear or branched alkylene or alkenylene chain, each of which substituents is unsubstituted unless otherwise specified.

[0352] "Alkoxyl" refers to a radical bonded through an oxygen atom of the formula -O-alkyl (where alkyl is the alkyl chain defined above).

[0353] "Alkenyl" refers to a linear or branched hydrocarbon chain radical group consisting only of carbon and hydrogen atoms, containing at least one carbon-carbon double bond, and having 2 to 12 carbon atoms. In certain embodiments, the alkenyl contains 2 to 8 carbon atoms. In other embodiments, the alkenyl contains 2 to 4 carbon atoms. The alkenyl is bonded to the rest of the molecule by a single bond, such as ethenyl (i.e., vinyl), propa-1-enyl (i.e., allyl), buta-1-enyl, penta-1-enyl, penta-1,4-dienyl, etc. Unless otherwise specifically stated herein, the alkenyl group is optionally substituted by one or more of the following substituents: halo, cyano, nitro, oxo, thioxo, imino, oxymo, trimethylsilanyl, -OR a , -SR a -OC(O)-R a , -N(R a )2, -C(O)R a , -C(O)OR a ,-C(O)N(R a )2, -N(R a )C(O)OR a -OC(O)-N(R a )2, -N(R a )C(O)R a , -N(R a )S(O) t R a (wherein t is 1 or 2), -S(O) t Ure a (wherein t is 1 or 2), -S(O) t R a (wherein the formula t is 1 or 2) and -S(O) t N(R a )2 (wherein t is 1 or 2) (wherein each R aThese are independently hydrogen, alkyl (optionally substituted with halogen, hydroxy, methoxy, or trifluoromethyl), fluoroalkyl, carbocyryl (optionally substituted with halogen, hydroxy, methoxy, or trifluoromethyl), carbocyrylalkyl (optionally substituted with halogen, hydroxy, methoxy, or trifluoromethyl), aryl (optionally substituted with halogen, hydroxy, methoxy, or trifluoromethyl), aralkyl (optionally substituted with halogen, hydroxy, methoxy, or trifluoromethyl), heterocyclyl (optionally substituted with halogen, hydroxy, methoxy, or trifluoromethyl), heterocyclylalkyl (optionally substituted with halogen, hydroxy, methoxy, or trifluoromethyl), heteroaryl (optionally substituted with halogen, hydroxy, methoxy, or trifluoromethyl), or heteroarylalkyl (optionally substituted with halogen, hydroxy, methoxy, or trifluoromethyl).

[0354] "Alkynyl" refers to a linear or branched hydrocarbon chain radical group consisting only of carbon and hydrogen atoms, containing at least one carbon-carbon triple bond, and having 2 to 12 carbon atoms. In certain embodiments, the alkynyl contains 2 to 8 carbon atoms. In other embodiments, the alkynyl contains 2 to 6 carbon atoms. In other embodiments, the alkynyl contains 2 to 4 carbon atoms. The alkynyl is bonded to the rest of the molecule by a single bond, such as ethynyl, propynyl, butynyl, pentynyl, hexynyl, etc. Unless otherwise specifically stated herein, the alkynyl group is optionally substituted by one or more of the following substituents: halo, cyano, nitro, oxo, thioxo, imino, oxymo, trimethylsilanyl, -OR a , -SR a -OC(O)-R a , -N(R a )2, -C(O)R a , -C(O)OR a ,-C(O)N(R a )2, -N(Ra )C(O)OR a -OC(O)-N(R a )2, -N(R a )C(O)R a , -N(R a )S(O) t R a (wherein t is 1 or 2), -S(O) t Ure a (wherein t is 1 or 2), -S(O) t R a (wherein the formula t is 1 or 2) and -S(O) t N(R a )2 (wherein t is 1 or 2) (wherein each R a These are independently hydrogen, alkyl (optionally substituted with halogen, hydroxy, methoxy, or trifluoromethyl), fluoroalkyl, carbocyryl (optionally substituted with halogen, hydroxy, methoxy, or trifluoromethyl), carbocyrylalkyl (optionally substituted with halogen, hydroxy, methoxy, or trifluoromethyl), aryl (optionally substituted with halogen, hydroxy, methoxy, or trifluoromethyl), aralkyl (optionally substituted with halogen, hydroxy, methoxy, or trifluoromethyl), heterocyclyl (optionally substituted with halogen, hydroxy, methoxy, or trifluoromethyl), heterocyclylalkyl (optionally substituted with halogen, hydroxy, methoxy, or trifluoromethyl), heteroaryl (optionally substituted with halogen, hydroxy, methoxy, or trifluoromethyl), or heteroarylalkyl (optionally substituted with halogen, hydroxy, methoxy, or trifluoromethyl).

[0355] An "alkylene" or "alkylene chain" refers to a straight or branched divalent hydrocarbon chain consisting only of carbon and hydrogen, containing no unsaturated atoms, and having 1 to 12 carbon atoms, with the rest of the molecule linked to a radical group, such as methylene, ethylene, propylene, and n-butylene. Alkylene chains are bonded to the rest of the molecule through single bonds and to the radical group through single bonds. The bonding points of an alkylene chain to the rest of the molecule and to the radical group are through one carbon in the alkylene chain or any two carbons in the chain. In certain embodiments, the alkylene comprises 1 to 8 carbon atoms (e.g., C1-C8 alkylene), 1 to 7 carbon atoms (e.g., C1-C7 alkylene), 1 to 6 carbon atoms (e.g., C1-C6 alkylene), 1 to 5 carbon atoms (e.g., C1-C5 alkylene), 1 to 4 carbon atoms (e.g., C1-C4 alkylene), 1 to 3 carbon atoms (e.g., C1-C3 alkylene), or 1 to 2 carbon atoms (e.g., C1-C2 alkylene). In other embodiments, the alkylene comprises 1 carbon atom (e.g., C1 alkylene), 5 to 8 carbon atoms (e.g., C5-C8 alkylene), 2 to 5 carbon atoms (e.g., C2-C5 alkylene), or 3 to 5 carbon atoms (e.g., C3-C5 alkylene). Unless otherwise specified, alkylene chains are optionally substituted with one or more of the following substituents: halo, cyano, nitro, oxo, thioxo, imino, oxymo, trimethylsilanyl, -OR a , -SR a -OC(O)-R a , -N(R a )2, -C(O)R a , -C(O)OR a ,-C(O)N(R a )2, -N(R a )C(O)OR a -OC(O)-N(R a )2, -N(R a )C(O)R a , -N(R a )S(O) t R a (wherein t is 1 or 2), -S(O) t Ure a(wherein t is 1 or 2), -S(O) t R a (wherein the formula t is 1 or 2) and -S(O) t N(R a )2 (wherein t is 1 or 2) (wherein each R a These are independently hydrogen, alkyl (optionally substituted with halogen, hydroxy, methoxy, or trifluoromethyl), fluoroalkyl, carbocyryl (optionally substituted with halogen, hydroxy, methoxy, or trifluoromethyl), carbocyrylalkyl (optionally substituted with halogen, hydroxy, methoxy, or trifluoromethyl), aryl (optionally substituted with halogen, hydroxy, methoxy, or trifluoromethyl), aralkyl (optionally substituted with halogen, hydroxy, methoxy, or trifluoromethyl), heterocyclyl (optionally substituted with halogen, hydroxy, methoxy, or trifluoromethyl), heterocyclylalkyl (optionally substituted with halogen, hydroxy, methoxy, or trifluoromethyl), heteroaryl (optionally substituted with halogen, hydroxy, methoxy, or trifluoromethyl), or heteroarylalkyl (optionally substituted with halogen, hydroxy, methoxy, or trifluoromethyl).

[0356] An "alkenylene" or "alkenylene chain" refers to a linear or branched divalent hydrocarbon chain consisting only of carbon and hydrogen, containing at least one carbon-carbon double bond, and having 2 to 12 carbon atoms, linking the rest of the molecule to a radical group. Alkenylene chains are bonded to the rest of the molecule through single bonds and to the radical group through single bonds. In certain embodiments, the alkenylene contains 2 to 8 carbon atoms (e.g., C2-C8 alkenylene), 2 to 5 carbon atoms (e.g., C2-C5 alkenylene), 2 to 4 carbon atoms (e.g., C2-C4 alkenylene), or 2 to 3 carbon atoms (e.g., C2-C3 alkenylene). In other embodiments, the alkenylene contains 2 carbon atoms (e.g., C2 alkenylene). In other embodiments, the alkenylene contains 5 to 8 carbon atoms (e.g., C5-C8 alkenylene) or 3 to 5 carbon atoms (e.g., C3-C5 alkenylene). Unless otherwise specified, alkenylene chains are optionally substituted with one or more of the following substituents: halo, cyano, nitro, oxo, thioxo, imino, oxymo, trimethylsilanyl, -OR a , -SR a -OC(O)-R a , -N(R a )2, -C(O)R a , -C(O)OR a ,-C(O)N(R a )2, -N(R a )C(O)OR a -OC(O)-N(R a )2, -N(R a )C(O)R a , -N(R a )S(O) t R a (wherein t is 1 or 2), -S(O) t Ure a (wherein t is 1 or 2), -S(O) t R a (wherein the formula t is 1 or 2) and -S(O) t N(R a )2 (wherein t is 1 or 2) (wherein each R aThese are independently hydrogen, alkyl (optionally substituted with halogen, hydroxy, methoxy, or trifluoromethyl), fluoroalkyl, carbocyryl (optionally substituted with halogen, hydroxy, methoxy, or trifluoromethyl), carbocyrylalkyl (optionally substituted with halogen, hydroxy, methoxy, or trifluoromethyl), aryl (optionally substituted with halogen, hydroxy, methoxy, or trifluoromethyl), aralkyl (optionally substituted with halogen, hydroxy, methoxy, or trifluoromethyl), heterocyclyl (optionally substituted with halogen, hydroxy, methoxy, or trifluoromethyl), heterocyclylalkyl (optionally substituted with halogen, hydroxy, methoxy, or trifluoromethyl), heteroaryl (optionally substituted with halogen, hydroxy, methoxy, or trifluoromethyl), or heteroarylalkyl (optionally substituted with halogen, hydroxy, methoxy, or trifluoromethyl).

[0357] An "alkylene" or "alkylene chain" refers to a linear or branched divalent hydrocarbon chain consisting only of carbon and hydrogen, containing at least one carbon-carbon triple bond, and having 2 to 12 carbon atoms, linking the rest of the molecule to a radical group. The alkylene chain is bonded to the rest of the molecule through single bonds and to the radical group through single bonds. In certain embodiments, the alkylene contains 2 to 8 carbon atoms (e.g., C2-C8 alkylene), 2 to 5 carbon atoms (e.g., C2-C5 alkylene), 2 to 4 carbon atoms (e.g., C2-C4 alkylene), 2 to 3 carbon atoms (e.g., C2-C3 alkylene), or 2 carbon atoms (e.g., C2 alkylene). In other embodiments, the alkylene contains 5 to 8 carbon atoms (e.g., C5-C8 alkylene) or 3 to 5 carbon atoms (e.g., C3-C5 alkylene). Unless otherwise specifically stated herein, the alkynylene chains are optionally substituted with one or more of the following substituents: halo, cyano, nitro, oxo, thioxo, imino, oxymo, trimethylsilanyl, -ORa , -SR a -OC(O)-R a , -N(R a )2, -C(O)R a , -C(O)OR a ,-C(O)N(R a )2, -N(R a )C(O)OR a -OC(O)-N(R a )2, -N(R a )C(O)R a , -N(R a )S(O) t R a (wherein t is 1 or 2), -S(O) t Ure a (wherein t is 1 or 2), -S(O) t R a (wherein the formula t is 1 or 2) and -S(O) t N(R a )2 (wherein t is 1 or 2) (wherein each R a These are independently hydrogen, alkyl (optionally substituted with halogen, hydroxy, methoxy, or trifluoromethyl), fluoroalkyl, carbocyryl (optionally substituted with halogen, hydroxy, methoxy, or trifluoromethyl), carbocyrylalkyl (optionally substituted with halogen, hydroxy, methoxy, or trifluoromethyl), aryl (optionally substituted with halogen, hydroxy, methoxy, or trifluoromethyl), aralkyl (optionally substituted with halogen, hydroxy, methoxy, or trifluoromethyl), heterocyclyl (optionally substituted with halogen, hydroxy, methoxy, or trifluoromethyl), heterocyclylalkyl (optionally substituted with halogen, hydroxy, methoxy, or trifluoromethyl), heteroaryl (optionally substituted with halogen, hydroxy, methoxy, or trifluoromethyl), or heteroarylalkyl (optionally substituted with halogen, hydroxy, methoxy, or trifluoromethyl).

[0358] The term "aryl" refers to a radical derived from aromatic monocyclic or polycyclic hydrocarbon ring systems by removing a hydrogen atom from a ring carbon atom. Aromatic monocyclic or polycyclic hydrocarbon ring systems contain only hydrogen and 5 to 18 carbon atoms, and at least one of the rings in the ring system is completely unsaturated, i.e., they contain a cyclic, delocalized (4n+2)π-electron system according to Hückel's theory. Ring systems from which aryl groups are derived include, but are not limited to, benzene, fluorene, indan, indene, tetralin, and naphthalene. Unless otherwise specifically stated, the term "aryl" or the prefix "ar-" (for example, in "aralkyl") refers to alkyl, alkenyl, alkynyl, halo, fluoroalkyl, cyano, nitro, optionally substituted aryl, optionally substituted aralkyl, optionally substituted aralkenyl, optionally substituted aralquinyl, optionally substituted carbocyclyl, optionally substituted carbocyclylalkyl, optionally substituted heterocyclyl, optionally substituted heterocyclylalkyl, optionally substituted heteroaryl, optionally substituted heteroarylalkyl, -R b -OR a ,-R b -OC(O)-R a ,-R b -OC(O)-OR a ,-R b -OC(O)-N(R a )2, -R b -N(R a )2, -R b -C(O)R a ,-R b -C(O)OR a ,-R b -C(O)N(R a )2, -R b -OR c -C(O)N(R a )2, -R b -N(R a )C(O)OR a ,-R b -N(R a )C(O)R a ,-R b -N(Ra )S(O) t R a (wherein the formula t is 1 or 2), -R b -S(O) t R a (wherein the formula t is 1 or 2), -R b -S(O) t Ure a (wherein the formula t is 1 or 2) and -R b -S(O) t N(R a )2 (wherein t is 1 or 2) (wherein each R a R is independently hydrogen, alkyl (optionally substituted with halogen, hydroxy, methoxy, or trifluoromethyl), fluoroalkyl, cycloalkyl (optionally substituted with halogen, hydroxy, methoxy, or trifluoromethyl), cycloalkylalkyl (optionally substituted with halogen, hydroxy, methoxy, or trifluoromethyl), aryl (optionally substituted with halogen, hydroxy, methoxy, or trifluoromethyl), aralkyl (optionally substituted with halogen, hydroxy, methoxy, or trifluoromethyl), heterocyclyl (optionally substituted with halogen, hydroxy, methoxy, or trifluoromethyl), heterocyclylalkyl (optionally substituted with halogen, hydroxy, methoxy, or trifluoromethyl), heteroaryl (optionally substituted with halogen, hydroxy, methoxy, or trifluoromethyl), or heteroarylalkyl (optionally substituted with halogen, hydroxy, methoxy, or trifluoromethyl), and each R b R is independently, directly bonded, or a linear or branched alkylene or alkenylene chain. c It is intended to include any aryl radical (which is a linear or branched alkylene or alkenylene chain) optionally substituted by one or more substituents independently selected from the chain, and unless otherwise indicated, each of the substituents is unsubstituted.

[0359] "Aralkill" is formula -R c-aryl(in the formula, R) c This refers to a radical of an alkylene chain (e.g., methylene, ethylene, etc.) as defined above. The alkylene chain portion of the aralkyl radical may be substituted as described above for alkylene chains. The aryl portion of the aralkyl radical may be substituted as described above for aryl groups.

[0360] "Aralkenyl" is expressed by formula -R d - Refers to an aryl radical (where Rd is the alkenylene chain as defined above). The aryl portion of the aralkenyl radical is optionally substituted as described above for the aryl group. The alkenylene chain portion of the aralkenyl radical is optionally substituted as defined above for the alkenylene group.

[0361] "Aralkyl" refers to a radical of the formula -Re-aryl (where Re is the alkynylene chain as defined above). The aryl portion of the aralkylyl radical is optionally substituted as described above for the aryl group. The alkynylene chain portion of the aralkylyl radical is optionally substituted as defined above for the alkynylene chain.

[0362] "Aralcoxy" is the formula -OR c -aryl(in the formula, R) c A argyl radical refers to a radical bonded through an oxygen atom of an alkylene chain (e.g., methylene, ethylene, etc.) as defined above. The alkylene chain portion of the argyl radical is optionally substituted as described above for the alkylene chain. The aryl portion of the argyl radical is optionally substituted as described above for the aryl group.

[0363] "Biaryl" refers to a radical of the formula -Ar-Ar (where the two aryl groups are linked by a single bond). Biphenyl is an example of a biaryl radical.

[0364] "Carbocyclylalkyl" is a compound of the formula -Rc -Carbocyclyl (in the formula, R c This refers to the radical of an alkylene chain (as defined above). The alkylene chain and carbocyclyl radical are optionally substituted as defined above.

[0365] "Carbocyclyl alkenyl" is represented by formula -R c -Carbocyclyl (in the formula, R c This refers to the radical of an alkenylene chain (as defined above). The alkenylene chain and carbocykyl radical are optionally substituted as defined above.

[0366] "Carbocyclylalkynyl" is represented by formula -R c -Carbocyclyl (in the formula, R c This refers to the radical of the alkynylene chain (as defined above). The alkynylene chain and carbocykyl radical are substituted as may be, as defined above.

[0367] A "carbocyclylalkoxy" refers to a radical bonded through the oxygen atom of the formula -O-Rc-carbocyclyl (where Rc is the alkylene chain defined above). The alkylene chain and the carbocyclyl radical are optionally substituted as defined above.

[0368] "Halo" or "halogen" refers to bromo, chloro, fluoro, or iodo substituents.

[0369] "Cyano" refers to the group -CN.

[0370] "Oxo" refers to the base = O.

[0371] "Haloalkyl" refers to an alkyl radical having one or more halogen substituents. For example, haloalkyls include groups such as trifluoromethyl, 3-fluoro-2-chloropropyl, and 4-bromocyclohexyl.

[0372] "Haloalkylenyl" refers to an alkenyl radical having one or more halogen substituents. For example, haloalkenyls include groups such as 2,2-difluorovinyl and 4-chloro-2-butenyl.

[0373] "Heteroalkyl" refers to a radical of a saturated linear or branched alkyl chain in which at least one carbon atom in the chain is replaced by a heteroatom, such as O, S, or N. In some embodiments, the heteroalkyl can contain, for example, 1 to 12, 1 to 10, or 1 to 6 carbon atoms, C1 to C 12 Heteroalkyl, C1~C 10 These are called heteroalkyls, or C1-C6 heteroalkyls. In certain cases, a heteroalkyl group contains one, two, three, or four independently selected heteroatoms instead of the one, two, three, or four individual carbon atoms in the alkyl chain. Typical heteroalkyl groups include, for example, CH2CH2OCH3, -CH2CH2NHCH3, and -CH2CH2N(CH3)CH3.

[0374] A "heterocyclyl" refers to a stable 3- to 18-membered non-aromatic ring radical containing 2- to 12 carbon atoms and 1- to 6 heteroatoms selected from nitrogen, oxygen, and sulfur. Unless otherwise specified, heterocyclyl radicals are monocyclic, bicyclic, tricyclic, or tetracyclic ring systems, which may include condensed or bridging ring systems. The heteroatoms in a heterocyclyl radical are sometimes oxidized. One or more nitrogen atoms, if present, are sometimes quaternized. Heterocyclyl radicals are partially or completely saturated. Heterocyclyls are bonded to the rest of the molecule through any atom of the ring(s) or ring(s). Examples of such heterocyclyl radicals, though not limited to these, include dioxolanil, thienyl[1,3]dithianil, decahydroisoquinolyl, imidazolinil, imidazolidinil, isothiazolidinil, isoxazolidinil, morpholinil, octahydroindolyl, octahydroisoindolyl, 2-oxopiperazinil, 2-oxopiperidinil, 2-oxopyrrolidinil, oxazolidinil, piperidinil, piperazinil, 4-piperidonil, pyrrolidinil, pyrazolidinil, quinuclidinil, thiazolidinil, tetrahydrofuryl, trithianil, tetrahydropyranil, thiomorpholinil, thiamorpholinil, 1-oxo-thiomorpholinil, and 1,1-dioxo-thiomorpholinil. Unless otherwise specifically stated, the term "heterocyclyl" includes alkyl, alkenyl, alkynyl, halo, fluoroalkyl, oxo, thioxo, cyano, nitro, optionally substituted aryl, optionally substituted aralkyl, optionally substituted aralkenyl, optionally substituted aralquinyl, optionally substituted carbocyclyl, optionally substituted carbocyclylalkyl, optionally substituted heterocyclyl, optionally substituted heterocyclylalkyl, optionally substituted heteroaryl, optionally substituted heteroarylalkyl, -R b -OR a ,-R b -OC(O)-R a ,-R b -OC(O)-OR a ,-R b -OC(O)-N(Ra )2, -R b -N(R a )2, -R b -C(O)R a ,-R b -C(O)OR a ,-R b -C(O)N(R a )2, -R b -OR c -C(O)N(R a )2, -R b -N(R a )C(O)OR a ,-R b -N(R a )C(O)R a ,-R b -N(R a )S(O) t R a (wherein the formula t is 1 or 2), -R b -S(O) t R a (wherein the formula t is 1 or 2), -R b -S(O) t Ure a (wherein the formula t is 1 or 2) and -R b -S(O) t N(R a )2 (wherein t is 1 or 2) (wherein each R aR is independently hydrogen, alkyl (optionally substituted with halogen, hydroxy, methoxy, or trifluoromethyl), fluoroalkyl, cycloalkyl (optionally substituted with halogen, hydroxy, methoxy, or trifluoromethyl), cycloalkylalkyl (optionally substituted with halogen, hydroxy, methoxy, or trifluoromethyl), aryl (optionally substituted with halogen, hydroxy, methoxy, or trifluoromethyl), aralkyl (optionally substituted with halogen, hydroxy, methoxy, or trifluoromethyl), heterocyclyl (optionally substituted with halogen, hydroxy, methoxy, or trifluoromethyl), heterocyclylalkyl (optionally substituted with halogen, hydroxy, methoxy, or trifluoromethyl), heteroaryl (optionally substituted with halogen, hydroxy, methoxy, or trifluoromethyl), or heteroarylalkyl (optionally substituted with halogen, hydroxy, methoxy, or trifluoromethyl), and each R b R is independently, directly bonded, or a linear or branched alkylene or alkenylene chain. c The terms are intended to include a heterocyclyl radical defined on which is optionally substituted by one or more substituents selected from linear or branched alkylenes or alkenylenes, and unless otherwise indicated, each of the above substituents is unsubstituted.

[0375] An "N-heterocyclyl" or "N-bonded heterocyclyl" refers to a heterocyclyl radical as defined above, containing at least one nitrogen atom, where the bond site of the heterocyclyl radical to the rest of the molecule is through the nitrogen atom of the heterocyclyl radical. N-heterocyclyl radicals are optionally substituted as described above for heterocyclyl radicals. Examples of such N-heterocyclyl radicals, but not limited to, include 1-morpholinyl, 1-piperidinyl, 1-piperazinyl, 1-pyrrolidinyl, pyrazolidinyl, imidazolinyl, and imidazolidinyl.

[0376] "Heterocyclylalkyl" is a compound of formulas -R c -heterocycline (wherein R is R in the formula) c This refers to a radical of an alkylene chain (as defined above). If the heterocyclyl is a nitrogen-containing heterocyclyl, the heterocyclyl is optionally bonded to the alkyl radical at a nitrogen atom. The alkylene chain of the heterocyclylalkyl radical is optionally substituted as defined above for the alkylene chain. The heterocyclyl portion of the heterocyclylalkyl radical is optionally substituted as defined above for the heterocyclyl group.

[0377] "Heterocyclylalkoxy" is a formula -OR c -heterocycline (wherein R is R in the formula) c This refers to a radical bonded through the oxygen atom of an alkylene chain (as defined above). If the heterocyclyl is a nitrogen-containing heterocyclyl, the heterocyclyl is optionally bonded to the alkyl radical at the nitrogen atom. The alkylene chain of the heterocyclylalkoxy radical is optionally substituted as defined above for the alkylene chain. The heterocyclyl portion of the heterocyclylalkoxy radical is optionally substituted as defined above for the heterocyclyl group.

[0378] "Heteroaryl" refers to a radical derived from a 3- to 18-membered aromatic ring radical containing 2- to 17 carbon atoms and 1- to 6 heteroatoms selected from nitrogen, oxygen, and sulfur. As used herein, a heteroaryl radical is a monocyclic, bicyclic, tricyclic, or tetracyclic ring system in which at least one of the rings in the ring system is completely unsaturated, i.e., cyclic, delocalized (4n+2)π-electron system according to Hückel theory. Heteroaryls include condensed or bridging ring systems. The heteroatoms in the heteroaryl radical are optionally oxidized. One or more nitrogen atoms, if present, are optionally quaternized. The heteroaryl is bonded to the rest of the molecule through any atom of the ring(s)(or rings). Examples of heteroaryls are not limited to these, but include azepinyl, acridinyl, benzimidazolyl, benzoindolyl, 1,3-benzodioxolyl, benzofuranil, benzoxazolyl, benzo[d]thiazolyl, benzothiadiazolyl, benzo[b][1,4]dioxepinyl, benzo[b][1,4]oxazinyl, 1,4-benzodioxanil, benzonaphthofuranil, benzoxazolyl, benzodioxolyl, benzodioxynil, benzopyranil, benzopyranonil, benzofuranil, benzothienyl (benzothiophenyl), benzothieno[3,2-d]pyrimidinil, benzotriazolyl, benzo[4,6] Imidazon[1,2-a]pyridinyl, carbazolyl, cinnolinyl, cyclopenta[d]pyrimidinyl, 6,7-dihydro-5H-cyclopenta[4,5]thieno[2,3-d]pyrimidinyl, 5,6-dihydrobenzo[h]quinazolinyl, 5,6-dihydrobenzo[h]cinnolinyl, 6,7-dihydro-5H-benzo[6,7]cyclohepta[1,2-c]pyridadinyl, dibenzofuranyl, dibenzothiophenyl, furanyl, furanonyl, furo[3,2-c]pyridinyl, 5,6,7,8,9,10-hexahydrocycloocta[d]pyrimidinyl, 5,6,7,8,9,10-hexahydrocycloocta[d]pyrimidinyl, 5,6,7,8,910-Hexahydrocycloocta[d]pyridinyl, isothiazolyl, imidazolyl, indazolyl, indazolyl, isoindolyl, indolinyl, isoindolyl, isoquinolyl, indolidinyl, isoxazolyl, 5,8-methano-5,6,7,8-tetrahydroquinazolinyl, naphthilidinyl, 1,6-naphthilidinolyl, oxadiazolyl, 2-oxoazepi Nyl, oxazolyl, oxyranil, 5,6,6a,7,8,9,10,10a-octahydrobenzo[h]quinazolinyl, 1-phenyl-1H-pyrrolyl, phenadinyl, phenothiazinyl, phenoxadinyl, phthalazinyl, pteridinyl, purinyl, pyrrolyl, pyrazolyl, pyrazolo[3,4-d]pyrimidinyl, pyridinyl, pyrido[3,2-d]pyrimidinyl, pyrido[3,4 -d]pyrimidinyl, pyrazinyl, pyrimidinyl, pyridadinyl, pyrrolyl, quinazolinyl, quinoxalinyl, quinolinyl, isoquinolinyl, tetrahydroquinolinyl, 5,6,7,8-tetrahydroquinazolinyl, 5,6,7,8-tetrahydrobenzo[4,5]thieno[2,3-d]pyrimidinyl, 6,7,8,9-tetrahydro-5H-cyclohepta[4,5]thieno[2,3-d Examples include pyrimidinyl, 5,6,7,8-tetrahydropyrido[4,5-c]pyridazinyl, thiazolyl, thiadiazolyl, triazolyl, tetrazolyl, triazinyl, thieno[2,3-d]pyrimidinyl, thieno[3,2-d]pyrimidinyl, thieno[2,3-c]pyridinyl, and thiophenyl (i.e., thienyl). Unless otherwise specifically stated, the term "heteroaryl" includes alkyl, alkenyl, alkynyl, halo, fluoroalkyl, haloalkenyl, haloalkynyl, oxo, thioxo, cyano, nitro, optionally substituted aryl, optionally substituted aralkyl, optionally substituted aralkenyl, optionally substituted aralquinyl, optionally substituted carbocyclyl, optionally substituted carbocyclylalkyl, optionally substituted heterocyclyl, optionally substituted heterocyclylalkyl, optionally substituted heteroaryl, optionally substituted heteroarylalkyl, -R, b -ORa ,-R b -OC(O)-R a ,-R b -OC(O)-OR a ,-R b -OC(O)-N(R a )2, -R b -N(R a )2, -R b -C(O)R a ,-R b -C(O)OR a ,-R b -C(O)N(R a )2, -R b -OR c -C(O)N(R a )2, -R b -N(R a )C(O)OR a ,-R b -N(R a )C(O)R a ,-R b -N(R a )S(O) t R a (wherein the formula t is 1 or 2), -R b -S(O) t R a (wherein the formula t is 1 or 2), -R b -S(O) t Ure a (wherein the formula t is 1 or 2) and -R b -S(O) t N(R a )2 (wherein t is 1 or 2) (wherein each R aR is independently hydrogen, alkyl (optionally substituted with halogen, hydroxy, methoxy, or trifluoromethyl), fluoroalkyl, cycloalkyl (optionally substituted with halogen, hydroxy, methoxy, or trifluoromethyl), cycloalkylalkyl (optionally substituted with halogen, hydroxy, methoxy, or trifluoromethyl), aryl (optionally substituted with halogen, hydroxy, methoxy, or trifluoromethyl), aralkyl (optionally substituted with halogen, hydroxy, methoxy, or trifluoromethyl), heterocyclyl (optionally substituted with halogen, hydroxy, methoxy, or trifluoromethyl), heterocyclylalkyl (optionally substituted with halogen, hydroxy, methoxy, or trifluoromethyl), heteroaryl (optionally substituted with halogen, hydroxy, methoxy, or trifluoromethyl), or heteroarylalkyl (optionally substituted with halogen, hydroxy, methoxy, or trifluoromethyl), and each R b R is independently, directly bonded, or a linear or branched alkylene or alkenylene chain. c The terms are intended to include a heteroaryl radical defined on which (a linear or branched alkylene or alkenylene chain) is optionally substituted by one or more substituents selected from such chains, and unless otherwise indicated, each of the substituents is unsubstituted.

[0379] "N-heteroaryl" refers to a heteroaryl radical as defined above, which contains at least one nitrogen atom, and the bonding site of the heteroaryl radical to the rest of the molecule is through the nitrogen atom in the heteroaryl radical. N-heteroaryl radicals are optionally substituted as described above for heteroaryl radicals.

[0380] "C-heteroaryl" refers to the heteroaryl radical as defined above, where the bonding site of the heteroaryl radical to the rest of the molecule passes through the carbon atoms in the heteroaryl radical. The C-heteroaryl radical is optionally substituted as described above for heteroaryl radicals.

[0381] A "heteroarylalkyl" refers to a radical of the formula -Rc-heteroaryl (where Rc is the alkylene chain as defined above). If the heteroaryl is a nitrogen-containing heteroaryl, the heteroaryl is optionally bonded to the alkyl radical by a nitrogen atom. The alkylene chain of the heteroarylalkyl radical is optionally substituted as defined above for the alkylene chain. The heteroaryl portion of the heteroarylalkyl radical is optionally substituted as defined above for the heteroaryl group.

[0382] A "heteroarylalkoxy" refers to a radical bonded through the oxygen atom of the formula -O-Rc-heteroaryl (where Rc is the alkylene chain as defined above). If the heteroaryl is a nitrogen-containing heteroaryl, the heteroaryl is optionally bonded to the alkyl radical at the nitrogen atom. The alkylene chain of the heteroarylalkoxy radical is optionally substituted as defined above for the alkylene chain. The heteroaryl portion of the heteroarylalkoxy radical is optionally substituted as defined above for the heteroaryl group.

[0383] Pharmacokinetic properties refer to the characteristics that a living organism (e.g., the body of the person to whom the compound is administered) performs on a compound. These properties include absorption, bioavailability, distribution, metabolism, and elimination. Improvements in these properties refer to changes in the administered compound to values ​​that are more desirable for its pharmacokinetic properties. For example, higher bioavailability is usually considered an improvement in pharmacokinetic properties.

[0384] "Pharmacodynamic" properties refer to the characteristics a compound performs on a living organism (e.g., the body of the target to which the compound is administered). These properties may include receptor binding, agonism, antagonism, and interactions with carrier proteins. Improvements in these properties refer to changes to values ​​that are more desirable for the drug properties of the administered compound. For example, stronger binding to a target receptor is usually considered an improvement in pharmacodynamic properties.

[0385] As described herein, certain compounds in this disclosure may contain “optionally substituted” moieties. Generally, the term “substituted,” whether preceded by the term “optionally,” means that one or more hydrogens of the specified moiety are replaced with appropriate substituents. Unless otherwise specified, an “optionally substituted” group may have appropriate substituents at each substitutedable position of the group, and two or more positions in any given structure may be substituted with two or more substituents selected from the specified group, and the substituents may be the same or different at each position. The combinations of substituents that come to mind preferably result in the formation of a stable or chemically viable compound.

[0386] As used herein, the term “stable” means a compound that remains substantially unchanged when subjected to conditions that enable their generation, detection, and, in certain embodiments, their recovery, purification, and use for one or more purposes disclosed herein.

[0387] Suitable substituents for substituted alkyl, alkylene, haloalkyl, haloalkylene, heteroalkyl, heteroalkylene, carbocykryl, heterocycline, aryl, and heteroaryl groups include, but are not limited to, halogens, =O, CN, -OR c , -NR d R e , -S(O) k R c , -NR c S(O)2R c -S(O)2NR d Re , -C(=O)OR c , -OC(=O)OR c -OC(=O)R c , -OC(=S)OR c , -C(=S)OR c -O(C=S)R c -C(=O)NR d R e , -NR c C(=O)R c -C(=S)NR d R e , -NR c C(=S)R c , -NR c (C=O)OR c -O(C=O)NR d R e , -NR c (C=S)OR c -O(C=S)NR d R e , -NR c (C=O)NR d R e , -NR c (C=S)NR d R e -C(=S)R c -C(=O)R c , C1~C6 alkyl, C1~C6 haloalkyl, C1~C6 heteroalkyl, carbocyryl, (C1~C6-alkylene)-carbocyryl, (C1~C6-heteroalkylene)-carbocyryl, heterocyclyl, (C1~C6-alkylene)-heterocyclyl, (C1~C6-heteroalkylene)-heterocyclyl, aryl, (C1~C6-alkylene)-aryl, (C1~C6-heteroalkylene)-aryl, heteroaryl, (C1~C6-alkylene)-heteroaryl, or (C1~C6-heteroalkylene)-heteroaryl (wherein each of the alkyl, alkylene, heteroalkyl, heteroalkylene, carbocyryl, heterocyclyl, aryl and heteroaryl is a halogen, OR c -NO2, -CN, -NR c C(=O)R c , -NR d R e , -S(O)k R c , -C(=O)OR c -C(=O)NR d R e -C(=O)R c , optionally substituted with one or more C1-C6 alkyl, C1-C6 haloalkyl, or C1-C6 heteroalkyl, R c R is hydrogen, hydroxyl, C1-C6 alkyl, C1-C6 heteroalkyl, carbocyryl, (C1-C6-alkylene)-carbocyryl, (C1-C6-heteroalkylene)-carbocyryl, heterocyclyl, (C1-C6-alkylene)-heterocyclyl, (C1-C6-heteroalkylene)-heterocyclyl, aryl, (C1-C6-alkylene)-aryl, (C1-C6-heteroalkylene)-aryl, heteroaryl, (C1-C6-alkylene)-heteroaryl, or (C1-C6-heteroalkylene)-heteroaryl, each of which may be substituted with one or more halogen, hydroxyl, C1-C6 alkyl, C1-C6 haloalkyl, C1-C6 heteroalkyl, carbocyryl, heterocyclyl, aryl, or heteroaryl; R d and R e k is independently selected from hydrogen, C1-C6 alkyl, or C1-C6 heteroalkyl (where k is 0, 1, or 2).

[0388] As used herein, the term “radical” refers to a fragment of a molecule that has an open valence, which is a bonding site for bond formation. A monovalent radical has one open valence so that it can form one bond with another chemical group. In some embodiments, the radicals of molecules used herein (e.g., radicals of folate acceptor binders) are created by removing one hydrogen atom from the molecule to create a monovalent radical with one open valence at the position where the hydrogen atom was removed. Where appropriate, the radical may be divalent, trivalent, etc., with two, three or more hydrogen atoms removed to create a radical that can bond to two, three or more chemical groups. Where appropriate, the open valence of a radical can be created by removing non-hydrogen atoms (e.g., halogen atoms) or two or more atoms (e.g., hydroxyl groups), as long as the atoms removed are part of the total atoms in the molecule forming the radical (no more than about 20% of the total number of atoms).

[0389] In some embodiments, the compounds disclosed herein contain one or more chiral centers, thus resulting in enantiomers, diastereomers, and other stereoisomeric forms defined with respect to absolute stereochemistry as (R)- or (S)-. Unless otherwise specified, all stereoisomeric forms of the compounds disclosed herein are intended to be contemplated by this disclosure. Where the compounds described herein contain an alkene double bond, unless otherwise specified, this disclosure is intended to include both E and Z geometric isomers (e.g., cis or trans). Similarly, all possible isomers, as well as their racemic and optically pure forms and all tautomeric forms, are also intended to be included. The term “geometric isomer” refers to the E or Z geometric isomers (e.g., cis or trans) of an alkene double bond. The term “positional isomer” refers to structural isomers around a central ring, e.g., ortho-, meta-, and para-isomers around a benzene ring. [Examples]

[0390] All solvents, reagents, and starting materials were purchased from commercial suppliers and, unless otherwise specified, were used without further purification or synthesized according to the methods described herein or using literature procedures.

[0391] Cysteine ​​Wang resin was obtained from Novabiochem (San Diego, California). Amino acids were purchased from Chem-Impex International (Chicago, Illinois). All reagents used in the synthesis were purchased from Sigma-Aldrich (St. Louis, Missouri). BromoPEG compounds were purchased from Broadpharm (San Diego, California). All other cell culture reagents, syringes, and disposable supplies were purchased from VWR (Chicago, Illinois).

[0392] Unless otherwise specified, all reactions were carried out at room temperature.

[0393] NMR spectra were recorded using a 500 MHz Bruker AV500HD spectrometer. All preparative high-performance liquid chromatography (HPLC) was performed on an Agilent 1200 Instrument using a reversed-phase XBridge OBD preparative column (19 × 150 mm, 5 μm) manufactured by Waters (Milford, Massachusetts), with UV detection at 254 nm. Low-resolution mass spectrometry-liquid chromatography / mass spectrometry (LRMS-LC / MS) was performed on an Agilent 1220 Infinity LC using a reversed-phase XBridge Shield RP18 column (3.0 × 50 mm, 3.5 μm). Compounds were purified using CombiFlash column chromatography. The purity of all final compounds was 95% or higher when determined by analytical HPLC on a reversed-phase column using binary ammonium acetate (20 mM, pH-7) and acetonitrile as eluents.

[0394] [Example 1] Synthesis of TLR7 agonist (compound 1) Compound 1 was synthesized according to the following scheme 2:

[0395] [ka]

[0396] Step 1: Synthesis of 2,2-dimethyl-3-((3-nitroquinoline-4-yl)amino)propan-1-ol: To a stirred solution of 4-chloro-3-nitroquinoline (1.2 equivalents) (compound 1' in Scheme 2) in N,N-dimethylformamide (10 mL), 3-amino-2,2-dimethylpropan-1-ol (1.5 equivalents) (compound 2' in Scheme 2) and triethylamine (2 equivalents) were added. The reaction mixture was heated at 70°C for 60 minutes and monitored by liquid chromatography-mass spectrometry (LCMS). It was then cooled, diluted with water, and stirred for a further 15 minutes. The precipitated solid was filtered and washed with water. The solid was dried under vacuum, yielding the substance (compound 3' in Scheme 2) as a yellow solid. The yield was 80%.

[0397] Step 2: Synthesis of 3-((3-aminoquinoline-4-yl)amino)-2,2-dimethylpropane-1-ol: 2,2-dimethyl-3-(3-nitroquinoline-4-ylamino)propan-1-ol (1 g) (compound 3' in Scheme 2) was dissolved in methanol (15 mL) and reduced for 4 hours on Pd / C (100 mg, 10 mol%) as a catalyst under hydrogen balloon conditions. The solution was then filtered and evaporated under reduced pressure to obtain 3-(3-aminoquinoline-4-ylamino)-2,2-dimethylpropan-1-ol (compound 4' in Scheme 2).

[0398] Step 3: Synthesis of N-(4-((3-hydroxy-2,2-dimethylpropyl)amino)quinoline-3-yl)pentanamide: Triethylamine (2 equivalents) and valeryl chloride (compound 5' in Scheme 2, 1.5 equivalents) were added to a stirred solution of compound 4' (1 g) in anhydrous tetrahydrofuran (THF) (10 mL). The reaction mixture was then stirred for 4 hours, followed by removal of the solvent under reduced pressure. The crude residue was dissolved in ethyl acetate (EtOAC), washed with water and brine, and dried over sodium sulfate. The combined organic layer was evaporated to dryness under vacuum to obtain intermediate amide compound 6 (see Scheme 2). The overall yield was 70%.

[0399] Step 4: Synthesis of 3-(2-butyl-1H-imidazo[4,5-c]quinoline-1-yl)-2,2-dimethylpropane-1-ol: To a stirred solution of amide compound 6 (1 g) in MeOH (15 mL), excess calcium oxide (10 equivalents) was added, and the solution was heated at 110°C for 96 hours. The solvent was then removed under vacuum, and the residue was purified using flash column chromatography (MeOH / dichloromethane mobile phase) to obtain compound 7 of Scheme 2. The yield was 60%.

[0400] Step 5: Synthesis of 3-(4-amino-2-butyl-1H-imidazo[4,5-c]quinoline-1-yl)-2,2-dimethylpropane-1-ol (compound 1): 3-Chloroperbenzoic acid (1.5 equivalents) was added to a stirred solution of compound 7 (100 mg) from Scheme 2 in anhydrous dichloromethane (1 mL), and the solution was refluxed at 50°C for 30 minutes. Once the starting material was completely consumed, the solvent was evaporated to dryness under vacuum.

[0401] Next, the residue was redissolved in anhydrous dichloromethane (1 mL), followed by the addition of trichloroacetyl isocyanate (2.0 equivalents) (see compound 9' in Scheme 2), and the reaction mixture was heated at 45°C for 30 minutes. After the reaction was complete, the solvent was removed under vacuum, the residue was redissolved in anhydrous MeOH (1 mL), followed by the addition of 25% methanolic sodium methoxide solution (0.2 mL). This was then heated at 75°C for 1 hour and allowed to cool. After removing the solvent under vacuum, the residue was purified using column chromatography (MeOH / dichloromethane) to obtain compound 1 (TLR7-1A) as a colorless liquid. The overall yield was 70%. LCMS: [M+H]+m / z=327.

[0402] [Example 2] Synthesis of TLR7 agonist (compound 2) Compound 2 was synthesized according to the following scheme 3:

[0403] [ka]

[0404] The synthetic procedure for preparing tert-butyl(3-(4-amino-2-butyl-1H-imidazo[4,5-c]quinoline-1-yl)-2,2-dimethylpropyl)carbamate (compound 16 in scheme 3) follows the procedure discussed above, using tert-butyl(3-amino-2,2-dimethylpropyl)carbamate as the starting material.

[0405] Boc-deprotection of tert-butyl(3-(4-amino-2-butyl-1H-imidazo[4,5-c]quinoline-1-yl)-2,2-dimethylpropyl)carbamate: To a stirred solution of tert-butyl(3-(4-amino-2-butyl-1H-imidazo[4,5-c]quinoline-1-yl)-2,2-dimethylpropyl) carbamate (100 mg, 1 equivalent) (compound 16 in Scheme 3) in 1,2-dichloromethane (1 mL), trifluoroacetic acid (10 equivalents) was added. This was stirred under a nitrogen atmosphere for 1 hour. Once the starting material was completely consumed as determined by thin-layer chromatography (TLC), it was evaporated to dryness under vacuum and basicized with saturated sodium bicarbonate. This was then purified using column chromatography (MeOH / dichloromethane) to obtain 1-(3-amino-2,2-dimethylpropyl)-2-butyl-1H-imidazo[4,5-c]quinoline-4-amine (compound 2-TLR7-1B) as a colorless liquid. LCMS: [M+H]+m / z=326.

[0406] [Example 3] Synthesis of TLR7 agonist (compound 3) Compound 3 was synthesized according to the following scheme 4:

[0407] [ka]

[0408] The synthetic procedure for preparing tert-butyl(3-(4-amino-2-butyl-1H-imidazo[4,5-c]quinoline-1-yl)-2,2-dimethylpropyl)carbamate (compound 23 in scheme 4) follows the procedure discussed above, using tert-butyl(1-amino-2-methylpropan-2-yl)carbamate (compound 17) as the starting material.

[0409] Boc-deprotection of tert-butyl(3-(4-amino-2-butyl-1H-imidazo[4,5-c]quinoline-1-yl)-2,2-dimethylpropyl)carbamate: Trifluoroacetic acid (10 equivalents) was added to a stirred solution of tert-butyl(3-(4-amino-2-butyl-1H-imidazo[4,5-c]quinoline-1-yl)-2,2-dimethylpropyl) carbamate (100 mg, 1 equivalent) (compound 23 in scheme 4) in 1,2-dichloromethane (1 mL). This was stirred under a nitrogen atmosphere for 1 hour. Once the starting material was completely consumed as determined by TLC, it was evaporated to dryness under vacuum and basicized with saturated sodium bicarbonate. This was then purified using column chromatography (MeOH / dichloromethane) to obtain 1-(2-amino-2-methylpropyl)-2-butyl-1H-imidazo[4,5-c]quinoline-4-amine (compound 3-TLR7-1C) as a colorless liquid. LCMS: [M+H]+m / z=312.

[0410] [Example 4] Synthesis of folic acid-NHS ester (compound 25): The synthesis of the releaseable TLR7-folate conjugate is described in Scheme 5:

[0411] [ka]

[0412] DMSO (20 mL), dicyclohexylcarbodiimide (0.94 g), triethylamine (0.5 mL), and N-hydroxysuccinimide (0.52 g) were added to a stirred solution of folic acid (1 g) (compound 24 in Scheme 5). This was stirred under a nitrogen atmosphere for approximately 12 hours. The solution was filtered to remove the dicyclohexylurea byproduct. Folic acid-NHS (compound 25) was precipitated with excess ethyl acetate, washed three times with anhydrous ether, dried under vacuum, and stored at -20°C. The yield was 60%.

[0413] [Example 5] Synthesis of non-releasing TLR7-folate conjugate-1 (compound 4): The synthesis of a non-releasing TLR7-folate conjugate (e.g., compound 4) is described in Scheme 6:

[0414] [ka]

[0415] (S)-1-(4-amino-2-butyl-1H-imidazo[4,5-c]quinoline-1-yl)-20-(4-(((2-amino-4-oxo-3,4-dihydropteridine-6-yl)methyl)amino)benzamide)-2,2-dimethyl-17-oxo-4,7,10,13-tetraoxa-16-azahenicosan-21-acid (compound 4; FA-PEG3-TLR7-1A non-release ("NR")) Step 1: tert-butyl(15-(2-butyl-1H-imidazo[4,5-c]quinoline-1-yl)-14,14-dimethyl-3,6,9,12-tetraoxapentadecyl)carbamate (compound 27 in scheme 6): NaH (2 equivalents) and N-Boc-PEG3-bromide (2 equivalents) (compound 26 in Scheme 6) were added to a stirred solution of intermediate hydroxyl compound 7 (500 mg) synthesized above (Scheme 2) in THF (5 mL). This was stirred under a nitrogen atmosphere for approximately 5 hours, after which the solvent was evaporated to dryness using a rotary evaporator. This was then quenched with water and diluted with dimethyl sulfoxide (DMSO). The crude reaction mixture was purified by HPLC using ammonium acetate and acetonitrile as mobile phases to obtain compound 27 (see Scheme 6) as a colorless liquid. The yield was 60%.

[0416] Step 2: tert-butyl(15-(4-amino-2-butyl-1H-imidazo[4,5-c]quinoline-1-yl)-14,14-dimethyl-3,6,9,12-tetraoxapentadecyl)carbamate (compound 28): 3-Chloroperbenzoic acid (1.5 equivalents) was added to a stirred solution of compound 27 (100 mg) in anhydrous dichloromethane (1 mL), and the solution was refluxed at 45°C for 30 minutes. Once the starting material was completely consumed, the solvent was evaporated to dryness under vacuum. The residue was then redissolved in anhydrous dichloromethane (1 mL), followed by the addition of trichloroacetyl isocyanate (2.0 equivalents), and the reaction mixture was heated at 45°C for 30 minutes.

[0417] After the reaction was complete, the solvent was removed under vacuum, the residue was redissolved in anhydrous MeOH (1 mL), and then 25% methanolic sodium methoxide solution (0.2 mL) was added. This was then heated at 75°C for 1 hour and cooled. The solvent was removed under vacuum, and the residue was purified using column chromatography (MeOH / dichloromethane) to obtain compound 28 as a colorless liquid. The overall yield was 40%.

[0418] Step 3: 1-(1-amino-14,14-dimethyl-3,6,9,12-tetraoxapentadecan-15-yl)-2-butyl-1H-imidazo[4,5-c]quinoline-4-amine: Trifluoroacetic acid (10 equivalents) was added to a stirred solution of intermediate compound 28 (100 mg) in dichloromethane (1 mL). This was stirred under a nitrogen atmosphere for about 1 hour, and the solvent was evaporated to dryness using a rotary evaporator. After completely removing the residual acid, the residue was dissolved in DMSO (1 mL) to obtain a DMSO solution.

[0419] Step 4: (S)-1-(4-amino-2-butyl-1H-imidazo[4,5-c]quinoline-1-yl)-20-(4-(((2-amino-4-oxo-3,4-dihydropteridine-6-yl)methyl)amino)benzamide)-2,2-dimethyl-17-oxo-4,7,10,13-tetraoxa-16-azahenicosan-21-acid (compound 4): To the DMSO solution from the previous step, folic acid-NHS (2.0 equivalents) (compound 25 from scheme 5) and N,N-diisopropylethylamine (2 equivalents) were added and the mixture was stirred for 60 minutes. The reaction was monitored via LC-MS. After the starting materials were completely consumed, the product was purified by HPLC using ammonium acetate and acetonitrile as the mobile phase, yielding product compound 4 as a yellow solid. The yield was 70%. The product was confirmed using LC-MS. The calculated mass for C46H60N12O9 was 924.46, and the measured mass [M+H]+ was 925.

[0420] [Example 6] Synthesis of folic acid-PEG3-cysteine ​​conjugate (compound 33): The synthesis of folic acid-PEG3-cysteine ​​conjugate (e.g., compound 33) is described in Scheme 7:

[0421] [ka]

[0422] The NH-Fmoc-Cys(Trt)-Wang resin (compound 29) was first deprotected using 20% ​​piperidine in DMF (3 × 10 mins). The free amine was then deprotected in the presence of benzotriazole-1-yloxytripyrrolidinophosphonium hexafluorophosphate (PyBop) (2 equivalents), diisopropylethylamine (DIPEA) (3 equivalents), and dimethylformamide (DMF). moc The resin was treated with PEG3-acid (1.5 equivalents) (compound 30). After the starting material was consumed, the resin was washed three times with DMF to obtain compound 31.

[0423] Next, this was deprotected using 20% ​​piperidine in DMF (3 × 10 minutes) and washed with DMF. The resulting free amine was treated with Fmoc-Glu(O) in the presence of PyBop (2 equivalents), DIPEA (3 equivalents), and DMF. tThe product was treated with Bu)-COOH (2 equivalents). The coupling product was deprotected with 20% piperidine in DMF (3 × 10 mins) and treated with pteroic acid (1.5 equivalents) in the presence of PyBop (2 equivalents), DIPEA (2 equivalents), and DMF. Once the coupling was complete and compound 32 was obtained, the trifluoroacetyl group was deprotected with a 50% ammonia-DMF solution (3 × 20 mins). Finally, the resin was cleaved using a TFA:TIPS:water:TCEP cocktail solution and purified by HPLC to obtain folate-PEG3-cysteine ​​(compound 33) as a yellow solid.

[0424] For HPLC purification, 20 mM ammonium acetate and acetonitrile at pH -5 were used as the mobile phase.

[0425] [Example 7] Synthesis of compounds 35 and 36: The synthesis of compounds 35 and 36 is described in Scheme 8:

[0426] [ka]

[0427] Synthesis of 3-(4-amino-2-butyl-1H-imidazo[4,5-c]quinoline-1-yl)-2,2-dimethylpropyl(2-(pyridine-2-yldisulfaneyl)ethyl) carbonate (compound 36 in scheme 8): A heterobifunctional linker (2 equivalents) (compound 34) was added to a solution of TLR7-1 (compound 1; 1 equivalent) and 4-dimethylaminopyridine (DMAP) (0.2 equivalents) in 1 mL of methylene chloride at room temperature under an N2 atmosphere. The mixture was then stirred at room temperature for 12 hours and purified by HPLC using ammonium acetate and methanol mobile phases to obtain compounds 35 and 36 of Scheme 8.

[0428] [Example 8] Synthesis of release TLR7-folate conjugate: The synthesis of a specific releasing TLR7-folate conjugate (e.g., compound 5) is described in Scheme 9:

[0429] [ka]

[0430] Folic acid-PEG3-cysteine ​​(compound 33, 1.2 equivalents) was added to a stirred solution of compound 36 (1 equivalent) in DMSO and stirred under a nitrogen atmosphere for approximately 1-2 hours. LC-MS analysis of the mixture showed the formation of compound 5 (FA-PEG3-TLR7-1A(Re)). The mixture was then purified by preparative HPLC using ammonium acetate and acetonitrile as mobile phases.

[0431] [Example 9] Synthesis of non-releasing TLR7-folate conjugate: The synthesis of a specific non-releasing TLR7-folate conjugate (e.g., compound 9) is described in Scheme 10:

[0432] [ka]

[0433] Synthesis of (9H-fluoren-9-yl)methyl(16-(4-amino-2-butyl-1H-imidazo[4,5-c]quinoline-1-yl)-15,15-dimethyl-12-oxo-3,6,9-trioxa-13-azahexadecyl)carbamate: Fmoc-N-amide PEG3 acid (1.5 equivalents), PyBop (2 equivalents), and DIPEA (2 equivalents) were added to a stirred solution of 1-(3-amino-2,2-dimethylpropyl)-2-butyl-1H-imidazo[4,5-c]quinoline-4-amine (compound 2; 1 equivalent) in DMF. This solution was stirred under a nitrogen atmosphere for approximately 2 hours, and the resulting mixture was purified using HPLC to obtain the indicated compound.

[0434] Step 2: Synthesis of (S)-1-(4-amino-2-butyl-1H-imidazo[4,5-c]quinoline-1-yl)-21-(4-(((2-amino-4-oxo-3,4-dihydropteridine-6-yl)methyl)amino)benzamide)-2,2-dimethyl-5,18-dioxo-8,11,14-trioxa-4,17-diazadocosan-22-acid (compound 9): Tris(2-aminoethyl)amine (10 equivalents) was added to a stirred solution of (9H-fluoren-9-yl)methyl(16-(4-amino-2-butyl-1H-imidazo[4,5-c]quinoline-1-yl)-15,15-dimethyl-12-oxo-3,6,9-trioxa-13-azahexadecyl)carbamate (1 equivalent) in THF. The mixture was stirred for approximately 1 hour, and the reaction was monitored by LC-MS. Once the starting material was completely consumed, the mixture was purified using HPLC. The purified sample (1 equivalent) was dissolved in DMSO and treated with folic acid-NHS (compound 25, scheme 5, 2 equivalents) and DIPEA (2 equivalents) for 2 hours. This was then purified using HPLC to obtain a non-releasing TLR7-folic acid conjugate (compound 9).

[0435] [Example 10] Effectiveness in human cells To evaluate the efficacy of TLR-7 agonists, compounds 1 (TLR7-1A), 2 (TLR7-1B), and 3 (TLR7-1C) were used to treat human peripheral blood mononuclear cells (PBMCs) for 24 hours, and cytokine production was then assessed. Human PBMCs were isolated from healthy donors by density gradient centrifugation following a standard procedure. 18 mL of blood was diluted with 2% fetal bovine serum (FBS) in phosphate-buffered saline (PBS) (1:1 dilution) and slowly transferred to a 50 mL SepMate® PBMC isolation tube containing 12 mL of Ficoll-Paque®. The tube was centrifuged at 1200 g for 10 minutes, and the PBMCs were transferred to a new 50 mL tube.

[0436] Wash PBMC in 2 × 50 mL of PBS with 2% FBS, then 2 × 10 5Individual cells were seeded in RPMI medium at a density of 200 μL per well plate. These were then treated with different concentrations of TLR7 agonist for 24 hours.

[0437] Compound A (TLR7-1) was used as a control. Cell culture supernatants were isolated, and cytokine levels were calculated using an enzyme-linked immunosorbent assay (ELISA) kit (Figure 2). As shown in Figure 2, TLR7 agonists (compounds 1, 2, and 3) resulted in increased IL-6 expression in PBMCs compared to the control.

[0438] When compounds 1, 2, and 3 were treated with human primary monocyte-derived M2 macrophages for 48 hours, these compounds efficiently induced interleukin-6 (IL-6) and CXC motif chemokine ligand 10 (CXCL-10) compared to the parent compound TLR7-1 (Figures 3A and 3B). Compounds 1, 2, and 3 polarize M2 macrophages into M1 macrophages, as indicated by increased M1 markers IL-6 (Figure 3A) and CXCL10 (Figure 3B).

[0439] [Example 11] Effectiveness in mice Female Balb / c mice were purchased from Charles Rivers, housed in a sterile environment with a standard 12-hour light-dark cycle, and maintained on a folate-deficient diet. All animal procedures were approved by the Purdue Board on Animal Experiments in accordance with National Institutes of Health guidelines.

[0440] Healthy mice were injected via tail vein with 10 nmol of compound A (TLR7-1) or compound 1 (TLR7-1A), and peripheral blood was collected at the indicated time points after drug injection (Figures 3C and 3D). The effects of the drugs on plasma levels of IL-6 (Figure 3C) and tumor necrosis factor alpha (TNF-α) (Figure 3D) were determined 1 or 1.5 hours after treatment. Both compound A and compound 1 stimulated systemic cytokine release in healthy mice.

[0441] [Example 12] Macrophage polarization Myeloid cells were isolated from the tibia and femur of male C57BL / 6 mice and differentiated into macrophages using mouse M-CSF (20 ng / mL). The macrophages were then polarized to the M2 phenotype for 48 hours using 20 ng / mL of IL-4 / IL-6 / IL-13. Interferon-γ (IFN-γ), interleukin-4 (IL-4), and interleukin-13 (IL-13) were obtained from Biolgend (San Diego, California), and lipopolysaccharide (LPS) was purchased from Sigma-Aldrich (St. Louis, Missouri).

[0442] Next, the obtained M2 macrophages were incubated with various concentrations of reciquimod, GSK2245035, compound 1, compound 2, and compound 3 for 48 hours. The supernatant was harvested and cytokines were analyzed using ELISA. (Reciquimod and GSK2245035 are known TLR7 agonists).

[0443] As shown in Figures 4A and 4B, and the increases in M1 markers IL-6 and TNF-α, compound 1 efficiently polarized macrophages from the M2 to the M1 phenotype.

[0444] [Example 13] Expression of FR-β and TLR7 in human M2 macrophages The M2 polarized macrophages obtained above were detached using Accutase cell detachment solution (Biolegend, San Diego, California) and gently lifted using a cell scraper. The cells were then washed with PBS and nonspecific binding was blocked by incubation with Fc receptor blocking solution (Biolegend, San Diego, California) for 10 minutes. The cells were then washed with PBS and resuspended in 150 μl of Cyto-Fast® Fix / Perm Buffer (Biolegend, San Diego, California). This was incubated at room temperature for 20 minutes and washed with 1 mL of 1× Cyto-Fast® perm wash solution. This was centrifuged at 350 g for 5 minutes, and the supernatant was discarded. A cocktail of TLR7 and folate receptor beta (FR-β) antibodies was prepared in 1× Cyto-Fast® Perm wash solution, and 100 mL of this was incubated with the cell suspension in the dark for 20 minutes. The cells were washed with 1 ml of perm wash solution and resuspended in staining buffer. These were used for flow cytometry and confocal microscopy imaging. Figures 5A and 5B confirm that M2 macrophages express TLR7 and FR-β, and Figure 6 shows that both TLR7 and FR-β co-localize in the same endosomes of the macrophages.

[0445] [Example 14] Disulfide cleavage test The stability kinetics of the folic acid-TLR7 conjugate were evaluated. Figure 7A shows FA-compound A (FA-TLR7-1), and Figure 7B shows compound 5 (releasing conjugate ("Re")) in the presence of thiols. The synthesis of FA-TLR7-1 is described in International Publication No. 2021007277, which is incorporated herein by reference in its entirety. Both conjugates were treated with dithiothreitol (DTT) (40 equivalents) in PBS. Samples were removed and analyzed by LC-MS at 0, 7, 30, and 50 minutes. As shown in Figures 7A and 7B, both conjugates were rapidly cleaved in the presence of DTT, releasing free drugs (e.g., immunomodulator; compound A) within 30 minutes.

[0446] Figure 8 shows a schematic diagram of the mechanisms of action of releasing and non-releasing folate-TLR7 conjugates. Releasing conjugates can release the free drug TLR7 agonist upon disulfide cleavage in endosomes. Since TLR7 and FR-β are present in the same endosome, non-releasing / non-cleavable conjugates can induce an immune response without cleavage. This avoids premature drug release, promotes the stability of the compound in circulation, increases its endosomal residence time, and ultimately may reduce the dose required to achieve therapeutic effect.

[0447] [Example 15] Efficacy test To evaluate the effectiveness of release and non-release conjugates, newly isolated PBMCs were subjected to a sampling rate of 1 million cells / cm³. 2 The monocytes were seeded in monocyte adhesion medium at the specified seeding density. This was incubated in an incubator at 5% CO2 and 37°C for approximately 2 hours. Non-adherent cells were removed, and the monocytes were washed three times with warm monocyte adhesion medium. The monocytes were cultured in folate-deficient RPMI1640 medium. These were differentiated into non-polarized macrophages by incubation in fresh RPMI medium containing 20 ng / mL M-CSF supplemented with 1% penicillin streptomycin (Invitrogen, Carlsbad, California) and 10% FBS.

[0448] Three days later, the culture medium was replaced with fresh RPMI medium containing 20 ng / mL of M-CSF. Then, on day 7, the obtained macrophages were polarized into M2 macrophages by incubation with 20 ng / mL of IL-4 and 20 ng / mL of IL-13 for two days.

[0449] Macrophages were detached using Accutase® cell detachment solution and seeded into 96-well plates at a density of 60,000 cells / well. To evaluate whether TLR7 agonists or their conjugates could reprogram M2-like macrophages to the M1 phenotype, cells were incubated with different concentrations of TLR7 agonists (compound A; TLR7-1) or their corresponding conjugates (e.g., FA-PEG3-TLR7-1A conjugate, releasing (compound 5) and non-releasing (compound 4)).

[0450] Different concentrations of TLR7 agonists or their folate conjugates (e.g., compounds 4 and 5) were incubated with the polarized M2-like macrophages for the indicated time. The culture medium was harvested to analyze secreted cytokines and to recover cells for qPCR analysis. Total RNA was collected for approximately 2 × 10⁶ using the Quick-RNA® MicroPerp Kit (Zymo Research, Irvine, California) according to the manufacturer's recommended protocol. 5 The RNA samples were isolated from macrophages. The RNA samples were then reverse transcribed into cDNA using a high-volume cDNA reverse transcription kit (Applied Biosystems, Foster City, California; No. 4368814).

[0451] qPCR analysis was performed using iTaq® Universal SYBR Green SuperMix (Bio-Rad Laboratories, Inc., Hercules, California; 1725121), an iCycler thermocycler, and iCycler iQ 3.0 software (Bio-Rad Laboratories Inc., Hercules, California) to track the expression of markers characteristic of macrophage polarization. IL-6 and TNF-α were used as markers for the M1 phenotype. Each sample was analyzed independently in triplicate for each marker. IL-6, CXCL-10, and TNF-α protein expression in cell culture supernatants was calculated using a commercially available ELISA kit (Biolegend, San Diego, California).

[0452] Figure 9 shows the results of treating human PMBC-derived M2 macrophages with compound 5 for 3+45 hours, demonstrating that treatment with compound 5 polarized M2 macrophages into M1 macrophages (evidence of an increase in M1 markers).

[0453] Figures 11A–11D show the results of compound 4 treatment of human PMBC-derived M2 macrophages for 3 hours or 3+45 hours. In the latter case, after 3 hours of incubation, the cell culture medium was replaced with fresh, drug-free medium and incubated for a further 45 hours. As supported by Figures 11A–11D, compound 4 polarized M2 macrophages into M1 macrophages (evidence of increased M1 markers).

[0454] [Example 16] Analysis of surface markers during macrophage polarization Human PBMC-derived M2 macrophages were treated with different concentrations of folic acid-TLR7 agonist (e.g., compound 4) for 48 hours. The macrophage cells were detached using Accutase cell detachment solution (Biolegend, San Diego, California; 423201) and gently lifted using a cell scraper. The cells were washed with PBS and nonspecific binding was blocked by incubation with Fc receptor blocking solution (Biolegend, San Diego, California) at room temperature for 10 minutes. The resulting single-cell suspensions were stained for the M2 macrophage marker CD206, as well as the M1 macrophage markers CD80 and CD40. All samples were then analyzed by flow cytometry.

[0455] As shown in Figures 12A and 12B, non-releasing conjugate compound 4 polarized M2 macrophages into M1 macrophages, as indicated by the increase in M1 surface markers CD40 and CD80.

[0456] [Example 17] In vivo efficacy trial Referring to Figure 13, 20 nmol / mouse folate-TLR7-1A conjugate (compound 4; FA-PEG3-TLR7-1A(NR)) was intravenously injected into healthy C57BL / 6-NCrl mice. Blood was collected at 10, 30, 60, 120, 180, 250, 360, 480, 600, 720, 1440, and 2880 minutes after drug injection. Blood was centrifuged at 1000 g for 10 minutes, and plasma was carefully collected into a clear tube. Acetonitrile was added to the plasma (acetonitrile / plasma sample = 2 / 1(v / v)), followed by the addition of an internal standard (0.5 ng of reciquimod). The mixture was thoroughly vortexed and centrifuged at 13,000 rpm for 10 minutes. The supernatant was collected and injected into Agilent 6410 NanoLC QQQ to obtain the pharmacokinetic analysis results shown in Figure 13.

[0457] In an additional in vivo study, 6-8 week old female BalB / c mice (Charles River Laboratories International, Inc., Wilmington, Massachusetts) were transferred to a folate-deficient diet (TD.95247, Envigo Corporation, Indianapolis, Indiana) upon arrival. On day 14, the mice were given 5.0 × 10⁶ 4 Individual 4T1 cells were subcutaneously transplanted, and the tumor was reduced to approximately 50 mm. 3 The mice were allowed to grow until they reached a certain size. Then, 100 μl of folate-TLR7 conjugate compound 4 or 10 nmol / mouse compound 5 were intravenously injected at different concentrations. The control group received 3% DMSO in 100 μl of PBS on the same schedule.

[0458] The tumor volume is calculated using the formula (a × b 2 The tumor size was measured simultaneously with a caliper using ) / 2 (where a is the maximum tumor diameter and b is the minimum tumor diameter) (see Figures 10A, 10B, and 14). If desired, mice were sacrificed and the resulting tumor fragments were dissociated using a human tumor dissociation kit. The resulting single-cell suspensions were stained for antibodies, and the samples were then analyzed by flow cytometry. Percentages of CD4 and CD8 T cell populations were tested in live cells isolated from 4T1 solid tumors in the untreated control group and the folate-TLR7 conjugate-treated group. Figures 10A–10E show the results of treatment with compound 5, and Figure 14 shows the results of treatment with compound 4. Both compounds resulted in a reduction in tumor size and shifted the polarization of macrophages from the M2 to the M1 phenotype.

[0459] An additional 4T1 solid tumor metastasis assay was also performed. In this assay, a lung metastasis colony formation assay was conducted according to the reported procedure (Cresswell et al., Folate receptor beta designates immunosuppressive tumor-associated myeloid cells that can be reprogrammed with folate-targeted drugs, Cancer Research 2021, 81(3), 671-684). 50,000 4T1 cells were transplanted into mammary fat pads of 6-8 week old Balb / c mice, and tumors were established at approximately 50 mm. 3 The cells were allowed to grow until they reached a certain size. The mice were then intravenously injected with 100 μl of folic acid-TLR7 conjugate (compound 4) (50 nmol / kg) once a week. The control group received an equal volume of PBS on the same schedule. At the end of the treatment, lung metastases were evaluated in the disease control and treatment groups by co-culturing lung digestion cells with 6-thioguanine (60 mmol / L) in complete RPMI-1640 medium in a petri dish for 10 days. Metastatic colonies were visualized by fixing the plate with 5 mL of methanol, followed by washing with 5 mL of deionized water, and then staining with 0.03% methylene blue (see Figure 15C). All blue colonies from each plate were then counted.

[0460] As shown in Figures 15A–15C, treatment with folic acid-TLR7 conjugate significantly reduced tumor volume and the number of metastatic colonies present in the subjects.

Claims

1. Compounds represented by formula (I): 【Chemistry 1】 or a pharma- ceutically acceptable salt thereof, R １ , R ３ , R ４ , R ５ are each independently H, an alkyl group, an alkoxy group, an alkenyl group, an alkynyl group, a cycloalkyl group, an aryl group, a halo group, a heteroaryl group, -COR ２ｘ , 【Chemistry 2】 and R ２ は、H、-OH、-NH ２ -NHR ２ｘ 、N ３ -NH-CH ２ -NH ２ 、-CONH ２ 、-SO ２ NH ２ -NH-CS-NH ２ 、 【Chemistry 3】 and Y is H, -OH, -NH ２ , -NHR ２ｘ , -O-R ２ｘ , -SO-R ２ｘ , -SH, -SO ３ H, -N ３ , -CHO, -COOH, -CONH ２ , -COSH, -COR ２ｘ , -SO ２ N.H. ２ , alkenyl, alkynyl, alkoxyl, -NH-CH ２ -NH ２ , -CONH ２ , -SO ２ N.H. ２ , -NH-CS-NH ２ , 【Chemistry 4】 and R ２ｘ and R ２ｙ Each of is H, —OH, —CH ２ -OH, -NH ２ , -CH ２ -NH ２ , -COOMe, -COOH, -CONH ２ , -COCH ３ independently selected from the group consisting of alkyl, alkenyl, alkynyl, cycloaliphatic, aryl, biaryl, and heteroaryl; Each R ２ｚ is -NH ２ , -NR ２ｑ R ２ｑ’ , -O-R ２ｑ , -SO-R ２ｑ and -COR ２ｑ are independently selected from the group consisting of: Each R ２ｑ and R ２ｑ’ is independently alkyl or H; 【Chemistry 5】 is a 3-10 membered N-containing non-aromatic monocyclic or bicyclic heterocycle; R ２１ is H or alkyl; n' is 0 to 30; In formula I, X １ , X ２ , X ３ Each of the groups independently represents CR ｑ or N, and each R ｑ is independently hydrogen, halogen, or optionally substituted alkyl; n is 0 to 30; m is 0 to 4; When n is 0, Y is H, —OH, or —O—R ２ｘ But not really).

2. Compounds represented by formula (IA): 【Chemistry 6】 or a pharma- ceutically acceptable salt thereof, R １ is an optionally substituted C ３ ~C ８ is alkyl; R ２ is H, -OR ｚ , -SO ２ N (R ｚ ) ２ , -NR ２ｘ R ２ｙ , or N ３ and Y is H, -OR ｚ , -NR ２ｘ R ２ｙ , -SR ｚ , -SOR ｚ , -SO ３ R ｚ , -N ３ , -COR ｚ , -COOR ｚ , -CON(R ｚ ) ２ , -COSR ｚ , -SO ２ N (R ｚ ) ２ , or -CON(R ｚ ) ２ and R ２ｘ and R ２ｙ are each independently hydrogen, -N(R ｚ ) ２ , -CON(R ｚ ) ２ , -C(R ｚ ) ２ -N(R ｚ ) ２ , -CS-N(R ｚ ) ２ or optionally substituted alkyl, and each R ｚ are independently hydrogen, halogen, or optionally substituted alkyl; R ２ｘ and R ２ｙ taken together form an optionally substituted heterocycloalkyl; Each R ３ are independently halogen, -N ３ , -CN, -NO ２ , -COR ｚ , -COOR ｚ , -CON(R ｚ ) ２ , -COSR ｚ , -SO ２ N (R ｚ ) ２ , or -CON(R ｚ ) ２ , alkyl, heteroalkyl, cycloalkyl, heterocycloalkyl, alkoxy, amino, hydroxy, or thiol, wherein said alkyl, alkoxy, heteroalkyl, cycloalkyl, or heterocycloalkyl is optionally substituted; R ４ and R ５ are each independently alkyl, alkoxy, halogen, or cycloalkyl, wherein said alkyl, alkoxy, and cycloalkyl are optionally substituted; n is 1 to 6; m is 0 to 4).

3. R １ Acyclic optionally substituted C ３ ~C ６ The compound of claim 1 or 2, wherein R 2 is alkyl and R 2 is -NR 2x R 2y or -NH 2 .

4. The following formula: 【Chemistry 7】 or a pharma- ceutically acceptable salt thereof.

5. The compound according to any one of claims 1, 2 and 4, wherein n is 1 to 3.

6. Y is -OH, OCH ３ , -NH ２ , -NHNH ２ , -NHCONH ２ , -SH, -SO ２ N.H. ２ , -N ３ , -COOH, -COCH ３ , -COOCH ３ or -CONH ２ 5. The compound of claim 1, 2, or 4,

7. R ４ and R ５ However, each independently, C １ ~C ４ 5. The compound of claim 1, 2, or 4, wherein:

8. The following formula: 【Chemistry 8】 or a pharma- ceutically acceptable salt thereof.

5. The compound of claim 1, 2, or 4, represented by one of:

9. Compound of formula (II) 【Chemistry 9】 or a pharma- ceutically acceptable salt thereof, R １ , R ３ , R ４ , R ５ are each independently H, an alkyl group, an alkoxy group, an alkenyl group, an alkynyl group, an alicyclic group, an aryl group, a biaryl group, a halo group, a heteroaryl group, -COR ２ｘ , 【Chemistry 10】 and R ２ は、H、-OH、-NH ２ -NHR ２ｘ 、N ３ -NH-CH ２ -NH ２ 、-CONH ２ 、-SO ２ NH ２ -NH-CS-NH ２ 、 【Chemistry 11】 and Z is a group of the formula GL-, GO-, GL-O-, GL-O-alkyl-, GL-S-, G-SO ２ -NH-,GL-NR ａ R ｂ -, GL-S(O) ｘ -alkyl-, G-L-CO-, G-L-aryl-, G-L-NH-CO-NH-, G-L-NH-O-, G-L-NH-NH-, G-L-NH-CS-NH, G-L-C(O)-alkyl-, G-L-SO ２ --, 【Chemistry 12】 Based on L is a linker and G is a folate receptor binding ligand; R ａ and R ｂ are each independently H, halo, hydroxy, alkoxy, aryl, amino, acyl, or C(O)R ｃ and R ｃ is alkyl, aryl, oxy or alkoxy; x is 0 to 3; R ２ｘ and R ２ｙ Each of is H, —OH, —CH ２ -OH, -NH ２ , -CH ２ -NH ２ , -COOMe, -COOH, -CONH ２ , -COCH ３ independently selected from the group consisting of alkyl, alkenyl, alkynyl, cycloaliphatic, aryl, biaryl, and heteroaryl; Each R ２ｚ is -NH ２ , -NR ２ｑ R ２ｑ’ , -O-R ２ｑ , -SO-R ２ｑ and -COR ２ｑ are independently selected from the group consisting of: Each R ２ｑ and R ２ｑ’ is independently alkyl or H; 【Chemistry 13】 is a 3-10 membered N-containing non-aromatic monocyclic or bicyclic heterocycle; R ２１ is H or alkyl; n' is 0 to 30; In formula II, X １ , X ２ , and X ３ are each independently ｑ or N, and each R ｑ are independently hydrogen, halogen, or optionally substituted alkyl; n is 0 to 30 and m is 0 to 4; When n is 0, Z is not attached to formula (II) by an oxygen atom.

10. The compound of claim 9, wherein the linker is a non-releasable linker.

11. The compound of claim 9, wherein the linker is a bivalent linker.

12. G is a group of formula (IV): 【Chemistry 14】 (Wherein, R is the following: 【Chemistry 15】 or any naturally occurring or unnatural amino acid or a derivative or fragment thereof) 10. The compound of claim 9 which is or contains the group:

13. G is a structure of formula (V) or formula (VI): 【Chemistry 16】 【Chemistry 17】 10. The compound of claim 9, which is a radical having the formula:

14. The structure: 【Chemistry 18】 10. The compound of claim 1, 2, or 9, represented by:

15. 15. A pharmaceutical composition comprising a compound according to any one of claims 1 to 14 and at least one pharma- ceutically acceptable excipient.

16. 15. A pharmaceutical composition comprising a therapeutically effective amount of one or more compounds according to any one of claims 1 to 14 and at least one pharma- ceutically acceptable excipient.

17. The pharmaceutical composition of claim 16 for treating cancer or a fibrotic disease or disorder in an individual.

18. The pharmaceutical composition of claim 17 for treating cancer.

19. 19. The pharmaceutical composition of claim 18, wherein the cancer is selected from the group consisting of lung cancer and breast cancer.

20. 18. The pharmaceutical composition of claim 17 for treating a fibrotic disease or disorder.

21. The pharmaceutical composition of claim 16 for converting a population of macrophages biased towards an M2-like phenotype to an M1-like phenotype, wherein the population of macrophages is present at a targeted location within an individual, and wherein the M2-like phenotype is associated with an anti-inflammatory / pro-fibrotic state, and the M1-like phenotype is associated with a pro-inflammatory / anti-fibrotic state.

22. The pharmaceutical composition described in claim 21, further for treating cancer.

23. 23. The pharmaceutical composition of claim 22, wherein the targeted location within the individual is a tumor microenvironment.

24. The pharmaceutical composition of claim 21, further for treating a fibrotic disease or disorder.

25. 25. A pharmaceutical composition according to any one of claims 17 to 24 for administering a second therapeutic agent.

26. The pharmaceutical composition of claim 25, further for treating a fibrotic disease or disorder, wherein the second therapeutic agent is an anti-inflammatory agent.

27. ​​The pharmaceutical composition of claim 25, further for treating cancer, wherein the second therapeutic agent is a chemotherapy agent or radiation therapy.

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