CN117295733A

CN117295733A - Immunomodulator and immunomodulator conjugates

Info

Publication number: CN117295733A
Application number: CN202280021195.0A
Authority: CN
Inventors: 大卫·M·弗格森; 彼得·古斯塔夫·拉森
Original assignee: University of Minnesota
Current assignee: University of Minnesota
Priority date: 2021-02-22
Filing date: 2022-02-22
Publication date: 2023-12-26

Abstract

The present disclosure provides compounds of formula I:

Description

Immunomodulator and immunomodulator conjugates

Cross Reference to Related Applications

The present application claims priority from U.S. provisional application No. 63/152,003, filed on 22 nd 2 nd year 2021, and U.S. provisional application No. 63/273,081, filed on 28 th 10 th 2021, which are incorporated herein by reference in their entirety.

Incorporated by reference

All publications, patents, and patent applications mentioned in this specification are herein incorporated by reference to the same extent as if each individual publication, patent, or patent application was specifically and individually indicated to be incorporated by reference.

Background

Toll-like receptor (TLR) modulators are useful in a variety of therapeutic settings, for example as vaccine adjuvants. However, many factors limit the efficacy of TLR modulating compounds such as imiquimod (imiquimod). As one example, while such compounds may induce pro-inflammatory cytokines, they may also simultaneously induce significant levels of anti-inflammatory cytokines, such as IL-10. Thus, there is an unmet need to develop TLR modulatory compounds that can trigger a more desirable ratio of pro-inflammatory cytokines to anti-inflammatory cytokines.

Disclosure of Invention

In some embodiments, TLR modulating compounds are disclosed that can trigger a more desirable ratio of pro-inflammatory cytokines to anti-inflammatory cytokines. In some cases, such TLR modulating compounds may be compounds of formula I:

(I)

wherein:

fused ring a is selected from the group consisting of:

R ¹ is H, (C) ₁ -C ₆ ) Alkyl, (C) ₂ -C ₆ ) Alkenyl group (C) ₂ -C ₆ ) Alkynyl, (C) ₁ -C ₆ ) Alkanoyl, (C) ₁ -C ₆ ) Alkoxycarbonyl, (C) ₁ -C ₆ ) Alkanoyloxy, (C) ₃ -C ₆ ) Cycloalkyl, aryl, heteroaryl, heterocycle, NR ^g R ^h Or R is ^m R ⁿ NC (=o) -, any of which (C) ₁ -C ₆ ) Alkyl, (C) ₂ -C ₆ ) Alkenyl group (C) ₂ -C ₆ ) Alkynyl, (C) ₁ -C ₆ ) Alkanoyl, (C) ₁ -C ₆ ) Incubating carbonyl group (C) ₁ -C ₆ ) Alkanoyloxy, (C) ₃ -C ₆ ) Cycloalkyl, aryl, heteroaryl, and heterocycle are optionally substituted with one or more groups independently selected from the group consisting of: halo, cyano, oxo, oxirane, (C) ₃ -C ₈ ) Cycloalkyl, aryl optionally substituted by alkyl, aryl substituted by carboxyl, heteroaryl, (C) ₁ -C ₆ ) Alkoxy, (C) ₁ -C ₆ ) Alkylthio, OR ^z 、-N(H)S(O) ₂ R ^r 、R ^s C(＝O)O-、-S-R ^w 、-NR ^x R ^y 、(C ₁ -C ₆ ) Alkoxycarbonyl and carboxyl, said alkyl being substituted by NR ^u R ^v Substitution;

R ² is H, (C) ₁ -C ₆ ) Alkyl, (C) ₂ -C ₆ ) Alkenyl group (C) ₂ -C ₆ ) Alkynyl, (C) ₁ -C ₆ ) Alkanoyl, (C) ₁ -C ₆ ) Alkoxycarbonyl, (C) ₁ -C ₆ ) Alkanoyloxy, (C) ₃ -C ₆ ) Cycloalkyl, aryl, heteroaryl, heterocycle, NR ^g R ^h Or R is ^m R ⁿ NC (=o) -, any of which (C) ₁ -C ₆ ) Alkyl, (C) ₂ -C ₆ ) Alkenyl group (C) ₂ -C ₆ ) Alkynyl, (C) ₁ -C ₆ ) Alkanoyl, (C) ₁ -C ₆ ) Alkoxycarbonyl, (C) ₁ -C ₆ ) Alkanoyloxy, (C) ₃ -C ₆ ) Cycloalkyl, aryl, heteroaryl, and heterocycle are optionally substituted with one or more groups independently selected from the group consisting of: hydroxy, halo, -SH, cyano, oxo, oxirane、(C ₃ -C ₈ ) Cycloalkyl, aryl, heteroaryl, (C) ₁ -C ₆ ) Alkoxy, (C) ₁ -C ₆ ) Alkylthio and NR ^g R ^h ；

R ³ Is H, halo, hydroxy, (C) ₁ -C ₆ ) Alkyl, (C) ₂ -C ₆ ) Alkenyl group (C) ₂ -C ₆ ) Alkynyl, (C) ₁ -C ₆ ) Alkanoyl, (C) ₁ -C ₆ ) Alkoxycarbonyl, (C) ₁ -C ₆ ) Alkanoyloxy, (C) ₃ -C ₆ ) Cycloalkyl, aryl, heteroaryl, heterocycle, NR ^g R ^h Or R is ^m R ⁿ NC (=o) -, any of which (C) ₁ -C ₆ ) Alkyl, (C) ₂ -C ₆ ) Alkenyl group (C) ₂ -C ₆ ) Alkynyl, (C) ₁ -C ₆ ) Alkanoyl, (C) ₁ -C ₆ ) Alkoxycarbonyl, (C) ₁ -C ₆ ) Alkanoyloxy, (C) ₃ -C ₆ ) Cycloalkyl, aryl, heteroaryl, and heterocycle are optionally substituted with one or more groups independently selected from the group consisting of: hydroxy, halo, -SH, cyano, oxo, oxirane, (C) ₃ -C ₈ ) Cycloalkyl, aryl, heteroaryl, (C) ₁ -C ₆ ) Alkoxy, (C) ₁ -C ₆ ) Alkylthio and NR ^g R ^h ；

R ⁴ Is R ^k -C(＝O)-、R ^k -O-C(＝O)-、R ^k -S(O) ₂ -O-、R ^c R ^d NC(＝O)-、R ^c R ^d NS(O) ₂ -、R ^e C(＝O)N(R ^e )-、R ^e S(O) ₂ NR ^f -, or at the 2-position by R ^k -C(＝O)-、R ^k -O-C(＝O)-、R ^k -S(O) ₂ -O-、R ^c R ^d NC(＝O)-、R ^c R ^d NS(O) ₂ -、R ^e C(＝O)N(R ^e ) -or R ^e S(O) ₂ NR ^f -substituted 1-ethylene;

R ^a is H, (C) ₁ -C ₆ ) Alkyl, (C) ₂ -C ₆ ) Alkenyl group (C) ₂ -C ₆ ) Alkyne (alkyne)Radical (C) ₁ -C ₆ ) Alkanoyl, (C) ₁ -C ₆ ) Alkoxycarbonyl, (C) ₃ -C ₆ ) Cycloalkyl, aryl or heteroaryl, any of which (C ₁ -C ₆ ) Alkyl, (C) ₂ -C ₆ ) Alkenyl group (C) ₂ -C ₆ ) Alkynyl, (C) ₁ -C ₆ ) Alkanoyl, (C) ₁ -C ₆ ) Alkoxycarbonyl, (C) ₃ -C ₆ ) Cycloalkyl, aryl, and heteroaryl are optionally substituted with one or more groups independently selected from the group consisting of: hydroxy, halo, -SH, cyano, oxirane, (C) ₃ -C ₈ ) Cycloalkyl, aryl, heteroaryl, (C) ₁ -C ₆ ) Alkoxy, (C) ₁ -C ₆ ) Alkylthio and NR ^g R ^h ；

R ^b Is H or X-Y;

each R ^c And R is ^d Independently H, (C) ₁ -C ₆ ) Alkyl, (C) ₂ -C ₆ ) Alkenyl group (C) ₃ -C ₈ ) Cycloalkyl, (C) ₃ -C ₈ ) Cycloalkyl (C) ₁ -C ₆ ) Alkyl, aryl (C) ₁ -C ₆ ) An aryl group; or together with the nitrogen to which they are attached form an aziridinyl, azetidinyl, morpholino, piperazino, pyrrolidino or piperidinyl ring, said ring being optionally substituted with one or more (C ₁ -C ₆ ) Alkyl substitution;

R ^e is H, (C) ₁ -C ₆ ) Alkyl or R ^aa ；

R ^f Is H or (C) ₁ -C ₆ ) An alkyl group;

each R ^g And R is ^h Independently H, (C) ₁ -C ₆ ) Alkyl, (C) ₂ -C ₆ ) Alkenyl group (C) ₃ -C ₈ ) Cycloalkyl, (C) ₃ -C ₈ ) Cycloalkyl (C) ₁ -C ₆ ) Alkyl, aryl (C) ₁ -C ₆ ) An alkyl group; or together with the nitrogen to which they are attached form an aziridinyl, azetidinyl, morpholino, piperazino, pyrrolidino or piperidinyl ring, said ring being capable of either Optionally by one or more (C ₁ -C ₆ ) Alkyl substitution;

R ^k is H, (C) ₁ -C ₆ ) Alkyl, (C) ₂ -C ₆ ) Alkenyl group (C) ₂ -C ₆ ) Alkynyl, (C) ₃ -C ₈ ) Cycloalkyl, trifluoromethyl, aryl or aryl (C) ₁ -C ₆ ) Alkyl groups, each of which (C ₁ -C ₆ ) The alkyl group being optionally substituted by one or more halo groups, (C) ₁ -C ₆ ) Alkanoyloxy, (C) ₁ -C ₆ ) Alkoxy, (C) ₃ -C ₈ ) Cycloalkyl substitution;

each R ^m And R is ⁿ Independently H, (C) ₁ -C ₆ ) Alkyl, (C) ₂ -C ₆ ) Alkenyl group (C) ₃ -C ₈ ) Cycloalkyl, (C) ₃ -C ₈ ) Cycloalkyl (C) ₁ -C ₆ ) Alkyl, aryl (C) ₁ -C ₆ ) An alkyl group; or together with the nitrogen to which they are attached form an aziridinyl, azetidinyl, morpholino, piperazino, pyrrolidino or piperidinyl ring, said ring being optionally substituted with one or more (C ₁ -C ₆ ) Alkyl substitution;

x is a linking group; and is also provided with

Y is a peptide, protein or maleimide;

wherein rings B and C in formula I can optionally be one or more independently selected from halo, hydroxy, nitro, (C) at one or more carbons ₁ -C ₆ ) Alkyl, (C) ₁ -C ₆ ) Alkenyl group (C) ₁ -C ₆ ) Alkynyl, (C) ₁ -C ₆ ) Alkoxy, (C) ₁ -C ₆ ) Alkanoyl, (C) ₁ -C ₆ ) Alkanoyloxy, (C) ₁ -C ₆ ) Alkoxycarbonyl, trifluoromethyl, trifluoromethoxy, cyano and NR ^p R ^q Further substitution of the groups of the group;

each R ^p And R is ^q Independently H or (C) ₁ -C ₆ ) An alkyl group; or together with the nitrogen to which they are attached form an aziridinyl, azetidinyl, morpholino, piperazino, A pyrrolo or piperidinyl ring, said ring being optionally substituted with one or more (C ₁ -C ₆ ) Alkyl substitution; and is also provided with

R ^r Is H, aryl or (C) optionally substituted by halo or aryl ₁ -C ₁₀ ) An alkyl group;

R ^s is H, aryl or (C) optionally substituted by halo or aryl ₁ -C ₁₀ ) An alkyl group;

each R ^u And R is ^v Independently H or (C) ₁ -C ₆ ) An alkyl group; or together with the nitrogen to which they are attached form an aziridinyl, azetidinyl, morpholino, piperazino, pyrrolidino or piperidinyl ring, said ring being optionally substituted with one or more (C ₁ -C ₆ ) Alkyl substitution;

R ^w is H, aryl or (C) optionally substituted by halo or aryl ₁ -C ₁₀ ) An alkyl group;

each R ^x And R is ^y Independently H or (C) ₁ -C ₆ ) An alkyl group; or together with the nitrogen to which they are attached form an aziridinyl, azetidinyl, morpholino, piperazino, pyrrolidino or piperidinyl ring, said ring being optionally substituted with one or more (C ₁ -C ₆ ) Alkyl substitution;

R ^z is H, aryl or (C) optionally substituted by halo or aryl ₁ -C ₁₀ ) An alkyl group; and is also provided with

R ^aa Is aryl optionally substituted with one or more groups independently selected from the group consisting of: hydroxy, halogen, nitro, cyano, (C) ₁ -C ₆ ) Alkoxy and optionally substituted with one or more halo (C ₁ -C ₆ ) An alkyl group;

or a pharmaceutically acceptable salt thereof.

In some embodiments, Y is an antigen.

The present disclosure also provides a pharmaceutical composition comprising a compound of formula I or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable diluent and/or carrier.

Further provided herein are methods for treating a pathological condition (e.g., a viral infection, bacterial infection, or cancer) in an animal (e.g., a mammal, such as a human) comprising administering to the animal a compound of formula I or a pharmaceutically acceptable salt thereof and/or a pharmaceutical composition comprising such a compound.

Further provided herein are methods for stimulating an immune response in an animal (e.g., a mammal, such as a human) comprising administering to the animal a compound of formula I or a pharmaceutically acceptable salt thereof and/or a pharmaceutical composition comprising such a compound.

Further provided herein are compounds of formula I, or pharmaceutically acceptable salts thereof, for use in the prophylactic and/or therapeutic treatment of a pathological condition (e.g., a viral infection, bacterial infection, or cancer) in an animal (e.g., a mammal, such as a human). Further provided herein are compounds of formula I, or pharmaceutically acceptable salts thereof, for use in medical therapy. The present disclosure also provides the use of a compound of formula I, or a pharmaceutically acceptable salt thereof, for the manufacture of a medicament useful for treating a pathological condition (e.g., a viral infection, bacterial infection, or cancer) in an animal (e.g., a mammal, such as a human).

The present disclosure further provides processes and intermediates disclosed herein that can be used to prepare compounds of formulas I-IV or pharmaceutically acceptable salts thereof.

Further provided herein are methods for synthesizing compounds of formulas I-IV and intermediates thereof, comprising performing the reactions according to any one of reaction schemes 1-5.

Detailed Description

I. Introduction to the invention

The present disclosure provides compounds (e.g., compounds of formulas I-IV) capable of interacting with one or more Toll-like receptors (TLRs), such as TLR-7, TLR-8, or a combination thereof. In some embodiments, the compounds of the present disclosure have agonistic effects on one or more TLRs. Thus, in some embodiments, the compounds provided herein can elicit an immune response in a subject, which can be used to treat a disease or pathological condition in a subject. Further provided herein are conjugates that can comprise a compound of the disclosure (e.g., one of formulas I-IV) coupled to another molecule. Such other molecules may be small molecules, peptides, proteins or nucleic acids. In some cases, conjugates herein comprise a compound of any one of formulas I-IV coupled to a peptide or protein. As further described herein, the compounds may be coupled (e.g., covalently coupled) to the peptide or protein via a linker.

Numerical ranges are understood to include inclusion, i.e., the indicated lower and upper limits. Furthermore, the term "about" as used herein generally refers to and encompasses ±10% of the indicated value unless specifically stated otherwise. For example, "about 10%" may indicate a range of 9% to 11%, while "about 1" may include a range of 0.9-1.1.

It should be noted that, as used herein and in the appended claims, the singular forms "a," "an," and "the" include plural referents unless the context clearly dictates otherwise. Thus, for example, reference to "a compound" includes a plurality of such compounds and equivalents thereof known to those skilled in the art, and so forth. Also, the terms "a" (or "an"), "one or more" and "at least one" can be used interchangeably herein. It should also be noted that the terms "comprising," "including," and "having" are used interchangeably.

As used herein, "comprising" is synonymous with "including," "containing," or "characterized by," and is inclusive or open-ended, and does not exclude additional, unrecited elements or method steps. As used herein, "consisting of … …" excludes any element, step or ingredient not specified in the claim elements. As used herein, "consisting essentially of … …" does not exclude materials or steps that do not materially affect the basic and novel characteristics of the claims.

As used herein, the term "subject" generally refers to an individual to whom a compound as described herein or a conjugate comprising such a compound is administered. Examples of "subjects" include, but are not limited to, mammals such as humans, rats, mice, guinea pigs, non-human primates, pigs, goats, cows, horses, dogs, cats, birds, and poultry. Typically, the subject is a rat, mouse, dog, non-human primate, or human. In some aspects, the subject is a human.

As used herein, the term "antibody" encompasses intact monoclonal antibodies, polyclonal antibodies, monospecific antibodies, multispecific antibodies (e.g., bispecific antibodies), including intact antibodies and antigen-binding antibody fragments, as well as reduced forms thereof in which one or more interchain disulfide bonds may be disrupted, which exhibit the desired biological activity, and provided that the antigen-binding antibody fragment has the desired number of attachment sites for the desired number of attachment groups such as the linker moieties described herein, if applicable. In aspects including compounds attached to antibodies by linkers, such linkers can be attached to the cysteine residues of the reduced interchain disulfide bonds and/or the sulfur atoms of the cysteine residues introduced by genetic engineering through succinimides or hydrolyzed succinimides. The natural form of an antibody is a tetramer and consists of two identical pairs of immunoglobulin chains, each pair having one light chain and one heavy chain. In each pair, the light and heavy chain variable domains (VL and VH) together are primarily responsible for binding to antigen. The light and heavy chain variable domains consist of framework regions interrupted by three hypervariable regions, also known as "complementarity determining regions" or "CDRs. The light and heavy chains also comprise constant regions that can be recognized by and interact with the immune system. The antibodies may be derived from any suitable species. In some aspects, the antibody is of human or murine origin, and in some aspects, the antibody is a human, humanized or chimeric antibody. Antibodies may be fucosylated or afucosylated to varying degrees.

An "antigen" is an entity that specifically binds to an antibody. Generally, as used herein, an "antigen" includes any substance that causes the immune system of an animal to produce antibodies or antigen-specific T cells against the substance. The term also includes haptens. The antigen may be a foreign substance in the environment, such as a chemical, bacteria, virus or pollen. Antigens may also be formed in vivo, such as with bacterial toxins, tissue cells, or tumor cells. An antigen is a molecular structure encoded by a substance against which an immune response is directed, such as a pathogen or tumor. Examples of antigens may be from pathogens such as bacteria or viruses (e.g., influenza, HIV or HCV). Alternatively, the antigen may be derived from a tumor cell or tumor cell lysate or synthetic peptide derived from a tumor or infectious organism. In one embodiment, the antigen comprises a peptide sequence comprising cysteine or lysine.

All terms, chemical names, expressions and names have the usual meaning known to a person skilled in the art. When a group of substituents is disclosed herein, it is understood that all individual members of the group and all subgroups, including any isomers, enantiomers, and diastereomers of the members of the group, are individually disclosed. When a Markush group (Markush group) or other grouping is used herein, all individual members of the group, as well as all possible combinations and subcombinations of the group, are intended to be individually included in this disclosure. When a compound is described herein, for example, a particular isomer, enantiomer or diastereomer of the compound is not specified in a chemical formula or chemical name, then the description is intended to include each isomer and enantiomer of the compound described separately or any combination thereof. In addition, unless otherwise indicated, all isotopic variations of the compounds disclosed herein are intended to be encompassed by the present disclosure. The specific names of compounds are intended to be exemplary, as it is known to one of ordinary skill in the art that the same compounds may be named differently.

The term "optionally substituted" refers to a substituted or unsubstituted designated group. As used herein, the term "substituted" refers to a compound (e.g., an alkyl chain) in which hydrogen is substituted with another reactive functional group or atom, as described herein.

As used herein, the term "group" may refer to a reactive functional group of a compound. The group of the compounds of the present invention refers to an atom or collection of atoms that are part of the compound. The groups of the present disclosure may be attached to other atoms of the compound by one or more covalent bonds. Groups may also be characterized in terms of their valency. The present disclosure includes groups characterized by monovalent, divalent, trivalent, etc. states.

As used herein, chemistryThe dashed line in the structure may be used to represent a bond to the rest of the molecule. For example, the number of the cells to be processed,is->For designating the 1-position as the point of attachment of the 1-methylcyclopentanoate to the rest of the molecule. Alternatively, for example +.>Is->May be used to indicate that a given moiety, in this example a cyclohexyl moiety, is attached to the molecule by a bond that is "capped" with a wavy line.

Unless otherwise indicated, the following definitions are used: halo is fluoro, chloro, bromo or iodo. Alkyl, alkoxy, alkenyl, alkynyl, and the like represent straight and branched groups; however, references to individual groups such as propyl include only straight chain groups, and specific references are to branched isomers such as isopropyl. Aryl represents phenyl or an ortho-fused bicyclic carbocyclyl having about nine to ten ring atoms, wherein at least one ring is aromatic. Heteroaryl groups include groups of monocyclic aromatic rings containing five or six ring atoms consisting of carbon and one to four heteroatoms, each heteroatom selected from the group consisting of non-peroxide oxygen, sulfur and N (X), where X is absent or is H, O, (C1-C4) alkyl, phenyl or benzyl, and groups of ortho-fused bicyclic heterocycles containing about eight to ten ring atoms of one to four heteroatoms, each heteroatom selected from the group consisting of non-peroxide oxygen, sulfur and N (X).

Those skilled in the art will appreciate that the compounds of the present disclosure having chiral centers may exist and be isolated in optically active and racemic forms. Some compounds may exhibit polymorphism. It is to be understood that the present disclosure encompasses any racemic, optically-active, polymorphic, or stereoisomeric form, or mixtures thereof, of a compound of the present disclosure, which possess the useful properties described herein, and how to prepare the optically-active form is well known in the art (e.g., by resolution of the racemic form by recrystallization techniques, by synthesis from an optically-active starting material, by chiral synthesis, or by chromatographic separation using a chiral stationary phase.

The particular values listed below for groups, substituents, and ranges are illustrative only; they do not exclude other defined values of groups and substituents or other values within defined ranges.

Specifically, (C) ₁ -C ₆ ) Alkyl can be methyl, ethyl, propyl, isopropyl, butyl, isobutyl, sec-butyl, pentyl, 3-pentyl or hexyl; (C) ₃ -C ₆ ) Cycloalkyl may be cyclopropyl, cyclobutyl, cyclopentyl or cyclohexyl; (C) ₃ -C ₆ ) Cycloalkyl (C) ₁ -C ₆ ) The alkyl group may be cyclopropylmethyl, cyclobutylmethyl, cyclopentylmethyl, cyclohexylmethyl, 2-cyclopropylethyl, 2-cyclobutylethyl, 2-cyclopentylethyl or 2-cyclohexylethyl; (C) ₁ -C ₆ ) Alkoxy can be methoxy, ethoxy, propoxy, isopropoxy, butoxy, isobutoxy, sec-butoxy, pentoxy, 3-pentoxy or hexoxy; (C) ₂ -C ₆ ) Alkenyl may be vinyl, allyl, 1-propenyl, 2-propenyl, 1-butenyl, 2-butenyl, 3-butenyl, 1-pentenyl, 2-pentenyl, 3-pentenyl, 4-pentenyl, 1-hexenyl, 2-hexenyl, 3-hexenyl, 4-hexenyl or 5-hexenyl; (C) ₂ -C ₆ ) Alkynyl may be ethynyl, 1-propynyl, 2-propynyl, 1-butynyl, 2-butynyl, 3-butynyl, 1-pentynyl, 2-pentynyl, 3-pentynyl, 4-pentynyl, 1-hexynyl, 2-hexynyl, 3-hexynyl, 4-hexynyl or 5-hexynyl; (C) ₁ -C ₆ ) Alkanoyl may be acetyl, propionyl or butyryl; (C) ₁ -C ₆ ) Alkoxycarbonyl may be methoxycarbonyl, ethoxycarbonyl, propoxycarbonyl, isopropoxycarbonyl, butoxycarbonyl, pentoxycarbonyl or hexoxycarbonyl; (C) ₂ -C ₆ ) Alkanoyloxy may be acetoxy, propionyloxy, butyryloxy, isobutyryloxy, pentanoyloxy or hexanoyloxyAn acyloxy group; aryl may be phenyl, indenyl or naphthyl; and heteroaryl may be furyl, imidazolyl, triazolyl, triazinyl, oxazolyl, isoxazolyl, thiazolyl, isothiazolyl, pyrazolyl, pyrrolyl, pyrazinyl, tetrazolyl, pyridyl (or N-oxide thereof), thienyl, pyrimidinyl (or N-oxide thereof), indolyl, isoquinolyl (or N-oxide thereof), or quinolinyl (or N-oxide thereof).

The term "alkyl" refers to an unsubstituted, straight or branched chain saturated hydrocarbon having the indicated number of carbon atoms (e.g., "C ₁ -C ₄ Alkyl "," C ₁ -C ₆ Alkyl "," C ₁ -C ₈ Alkyl "or" C ₁ -C ₁₀ "alkyl groups having 1 to 4, to 6, 1 to 8, or 1 to 10 carbon atoms, respectively) and are derived by removing one hydrogen atom from the parent alkane. Representative straight chain "C ₁ -C ₈ Alkyl "includes, but is not limited to, methyl, ethyl, n-propyl, n-butyl, n-pentyl, n-hexyl, n-heptyl, and n-octyl; and branched C ₁ -C ₈ Alkyl groups include, but are not limited to, isopropyl, sec-butyl, isobutyl, tert-butyl, isopentyl, and 2-methylbutyl.

The term "alkylene" refers to a divalent unsubstituted saturated branched or straight chain hydrocarbon having the indicated number of carbon atoms (e.g., C ₁ -C ₆ Alkylene has 1 to 6 carbon atoms) and has two monovalent centers derived by the removal of two hydrogen atoms from the same or two different carbon atoms of the parent alkane. Alkylene groups may be substituted with 1 to 6 fluoro groups, for example on a carbon backbone (e.g. -CHF-or-CF ₂ (-) or on a terminal carbon of a linear or branched alkylene group (such as-CHF) ₂ or-CF ₃ ). Alkylene groups include, but are not limited to: methylene (-CH) ₂ (-), ethylene (-CH) ₂ CH ₂ (-) n-propylene (-CH) ₂ CH ₂ CH ₂ -) subunit, n-propyl (-CH) ₂ CH ₂ CH ₂ (-), n-butyl (CH) ₂ CH ₂ CH ₂ CH ₂ (-), difluoro-methylene (-CF) ₂ (-), tetrafluoroethylene (-CF) ₂ CF ₂ (-), etc.

The term "alkenyl" refers to an unsubstituted, straight or branched hydrocarbon having at least one carbon-carbon double bond and a specified number of carbon atoms (e.g., "C ₂ -C ₈ Alkenyl "or" C ₂ -C ₁₀ "alkenyl groups have 2 to 8 or 2 to 10 carbon atoms, respectively). When the number of carbon atoms is not specified, the alkenyl group has 2 to 6 carbon atoms.

The term "heteroalkyl" refers to a stable, straight or branched chain saturated hydrocarbon having the indicated number of total atoms and at least one (e.g., 1 to 15) heteroatoms selected from the group consisting of O, N, si and S. The carbon and heteroatoms of the heteroalkyl groups may be oxidized (e.g., to form ketones, N-oxides, sulfones, etc.), and the nitrogen atom may be quaternized. The heteroatom may be located at any internal position of the heteroalkyl group and/or at any terminal end of the heteroalkyl group, including terminal ends of branched heteroalkyl groups), and/or at a position where the heteroalkyl group is attached to the remainder of the molecule. Heteroalkyl groups may be substituted with 1 to 6 fluoro groups, for example on the carbon backbone (e.g. -CHF-or-CF) ₂ (-) or on a terminal carbon of a straight or branched heteroalkyl group (such as-CHF) ₂ or-CF ₃ ). Examples of heteroalkyl groups include, but are not limited to, -CH ₂ -CH ₂ -O-CH ₃ 、-CH ₂ -CH ₂ -NH-CH ₃ 、-CH ₂ -CH ₂ -N(CH ₃ ) ₂ 、-C(＝O)-NH-CH ₂ -CH ₂ -NH-CH ₃ 、-C(＝O)-N(CH ₃ )-CH ₂ -CH ₂ -N(CH ₃ ) ₂ 、-C(＝O)-NH-CH ₂ -CH ₂ -NH-C(＝O)-CH ₂ -CH ₃ 、-C(＝O)-N(CH ₃ )-CH ₂ -CH ₂ -N(CH ₃ )-C(＝O)-CH ₂ -CH ₃ 、-O-CH ₂ -CH ₂ -CH ₂ -NH(CH ₃ )、-O-CH ₂ -CH ₂ -CH ₂ -N(CH ₃ ) ₂ 、-O-CH ₂ -CH ₂ -CH ₂ -NH-C(＝O)-CH ₂ -CH ₃ 、-O-CH ₂ -CH ₂ -CH ₂ -N(CH ₃ )-C(＝O)-CH ₂ -CH ₃ 、-CH ₂ -CH ₂ -CH ₂ -NH(CH ₃ )、-O-CH ₂ -CH ₂ -CH ₂ -N(CH ₃ ) ₂ 、-CH ₂ -CH ₂ -CH ₂ -NH-C(＝O)-CH ₂ -CH ₃ 、-CH ₂ -CH ₂ -CH ₂ -N(CH ₃ )-C(＝O)-CH ₂ -CH ₃ 、-CH ₂ -S-CH ₂ -CH ₃ 、-CH ₂ -CH ₂ -S(O)-CH ₃ 、-NH-CH ₂ -CH ₂ -NH-C(＝O)-CH ₂ -CH ₃ 、-CH ₂ -CH ₂ -S(O) ₂ -CH ₃ 、-CH ₂ -CH ₂ -O-CF ₃ and-Si (CH) ₃ ) ₃ . At most two heteroatoms may be consecutive, such as for example-CH ₂ -NH-OCH ₃ and-CH ₂ -O-Si(CH ₃ ) ₃ . Terminal polyethylene glycol (PEG) moieties are one type of heteroalkyl group.

The term "alkynyl" refers to an unsubstituted, straight or branched hydrocarbon having at least one carbon-carbon triple bond and a specified number of carbon atoms (e.g., "C ₂ -C ₈ Alkynyl "or" C ₂ -C ₁₀ "alkynyl groups have 2 to 8 or 2 to 10 carbon atoms, respectively). When the number of carbon atoms is not specified, the alkynyl group has 2 to 6 carbon atoms.

The term "acyl" refers to an alkyl, haloalkyl, alkenyl, alkynyl, arylcycloalkyl, heteroaryl, or heterocyclyl group as defined herein attached to the remainder of the compound through a c=o (carbonyl) group.

The term "carboxamide group" refers to a-C (=o) NRR 'group, wherein R and R' are independently selected from the group consisting of hydrogen, alkyl, alkenyl, alkynyl, arylcycloalkyl, heteroaryl, and heterocyclyl, as defined herein.

The term "heteroalkylene" refers to a divalent unsubstituted straight or branched chain group derived from a heteroalkyl group (e.g., as defined herein). Examples of heteroalkylene groups include, but are not limited to, -CH ₂ -CH ₂ -O-CH ₂ -、-CH ₂ -CH ₂ -O-CF ₂ -、-CH ₂ -CH ₂ -NH-CH ₂ -、-C(＝O)-NH-CH ₂ -CH ₂ -NH-CH ₂ --C(＝O)-N(CH ₃ )-CH ₂ -CH ₂ -N(CH ₃ )-CH ₂ -、-C(＝O)-NH-CH ₂ -CH ₂ -NH-C(＝O)-CH ₂ -CH ₂ -、-C(＝O)-N(CH ₃ )-CH ₂ -CH ₂ -N(CH ₃ )-C(＝O)-CH ₂ -CH ₂ -、-O-CH ₂ -CH ₂ -CH ₂ -NH-CH ₂ -、-O-CH ₂ -CH ₂ -CH ₂ -N(CH ₃ )-CH ₂ -、-O-CH ₂ -CH ₂ -CH ₂ -NH-C(＝O)-CH ₂ -CH ₂ -、-O-CH ₂ -CH ₂ -CH ₂ -N(CH ₃ )-C(＝O)-CH ₂ -CH ₂ -、-CH ₂ -CH ₂ -CH ₂ -NH-CH ₂ -、-CH ₂ -CH ₂ -CH ₂ -N(CH ₃ )-CH ₂ -、-CH ₂ -CH ₂ -CH ₂ -NH-C(＝O)-CH ₂ -CH ₂ -、-CH ₂ -CH ₂ -CH ₂ -N(CH ₃ )-C(＝O)-CH ₂ -CH ₂ -、-CH ₂ -CH ₂ -NH-C(＝O)-、-CH ₂ -CH ₂ -N(CH ₃ )-CH ₂ -、-CH ₂ -CH ₂ -N ⁺ (CH ₃ ) ₂ -、-NH-CH ₂ -CH ₂ (NH ₂ )-CH ₂ -and-NH-CH ₂ -CH ₂ (NHCH ₃ )-CH ₂ -. Divalent polyethylene glycol (PEG) moieties are one type of heteroalkylene group.

The term "alkoxy" refers to an alkyl group as defined herein attached to a molecule through an oxygen atom. For example, alkoxy groups include, but are not limited to, methoxy, ethoxy, n-propoxy, isopropoxy, n-butoxy, sec-butoxy, tert-butoxy, n-pentoxy, and n-hexoxy.

The term "alkylthio" refers to an alkyl group as defined herein attached to a molecule through a sulfur atom. For example, alkylthio groups include, but are not limited to, thiomethyl, thioethyl, thion-propyl, thioisopropyl, and the like.

The term "haloalkyl" refers to an unsubstituted, straight or branched, saturated hydrocarbon having the indicated number of carbon atoms (e.g., "C ₁ -C ₄ Alkyl "," C ₁ -C ₆ Alkyl "," C ₁ -C ₈ Alkyl "or" C ₁ -C ₁₀ "alkyl groups have 1 to 4, to 6, 1 to 8, or 1 to 10 carbon atoms, respectively), wherein at least one hydrogen atom of the alkyl group is replaced with a halogen (e.g., fluorine, chlorine, bromine, or iodine). When the number of carbon atoms is not specified, the haloalkyl group has 1 to 6 carbon atoms. Representative C _1-6 Haloalkyl includes, but is not limited to, trifluoromethyl, 2-trifluoroethyl, and 1-chloroisopropyl.

The term "cycloalkyl" refers to a cyclic, saturated or partially unsaturated hydrocarbon having the indicated number of carbon atoms (e.g., "C _3-8 Cycloalkyl "or" C _3-6 "cycloalkyl" has 3 to 8 or 3 to 6 carbon atoms, respectively). When the number of carbon atoms is not specified, the cycloalkyl group has 3 to 6 carbon atoms. Cycloalkyl includes bridged, fused and spiro ring systems, as well as bridged bicyclic systems, wherein one ring is aromatic and the other ring is unsaturated. Representative "C _3-6 Cycloalkyl "includes cyclopropyl, cyclobutyl, cyclopentyl and cyclohexyl.

The term "aryl" refers to an unsubstituted monovalent carbocyclic aromatic hydrocarbon group of 6 to 10 carbon atoms derived by removal of one hydrogen atom from a single carbon atom of the parent aromatic ring system. Aryl groups include, but are not limited to, phenyl, naphthyl, anthracenyl, biphenyl, and the like.

The term "heterocycle" refers to a saturated or partially unsaturated ring or multiple condensed ring system, including bridged, condensed, and spiro ring systems. Heterocycles can be described by the total number of atoms in the ring system, for example 3-10 membered heterocycles have a total of 3-10 ring atoms. The term includes mono-saturated or partially unsaturated rings (e.g., 3,4, 5, 6, or 7 membered rings) having about 1 to 6 carbon atoms and about 1 to 3 heteroatoms selected from the group consisting of oxygen, nitrogen, and sulfur in the ring. The ring may be substituted with one or more (e.g., 1,2, or 3) oxo groups, and the sulfur and nitrogen atoms may also be present in their oxidized forms. Such rings include, but are not limited to, azetidinyl, tetrahydrofuranyl, and piperidinyl. The term "heterocycle" also includes multiple fused ring systems (e.g., ring systems comprising 2,3, or 4 rings), wherein a single heterocycle (as defined above) may be fused with one or more heterocycles (e.g., decalinyl), carbocycles (e.g., decalinyl), or aryl groups. The rings of multiple fused ring systems may be linked to each other by fused bonds, spiro bonds and bridging bonds, as valence requirements allow. It will be appreciated that the attachment points of the plurality of fused ring systems (as defined above for heterocycles) may be located at any position of the plurality of fused ring systems, including the heterocyclic, aryl and carbocyclic portions of the ring. It is also understood that the attachment point of the heterocycle or heterocycle multiple condensed ring systems may be at any suitable atom of the heterocycle or heterocycle multiple condensed ring systems, including carbon atoms and heteroatoms (e.g., nitrogen). Exemplary heterocycles include, but are not limited to, aziridinyl, azetidinyl, pyrrolidinyl, piperidinyl, homopiperidinyl, morpholino, thiomorpholino, piperazino, tetrahydrofuranyl, dihydro oxazolyl, tetrahydropyranyl, tetrahydrothiopyranyl, 1,2,3, 4-tetrahydroquinolinyl, benzoxazinyl, dihydro oxazolyl, chromanyl, 1, 2-dihydropyridinyl, 2, 3-dihydrobenzofuranyl, 1, 3-benzodioxolyl, and 1, 4-benzodioxanyl.

The term "heteroaryl" refers to an aromatic hydrocarbon ring system having at least one heteroatom selected from the group consisting of O, N and S in a monocyclic or fused ring system. The ring or ring system has 4n+2 electrons in the conjugated pi system, where all atoms contributing to the conjugated pi system are in the same plane. In some embodiments, heteroaryl groups have a total of 5-10 ring atoms and 1, 2, or 3 heteroatoms (referred to as "5-10 membered heteroaryl"). Heteroaryl groups include, but are not limited to, imidazole, triazole, thiophene, furan, pyrrole, benzimidazole, pyrazole, pyrazine, pyridine, pyrimidine, and indole.

The term "hydroxy" refers to an-OH group. The term "cyano" refers to a-CN group. The term "carboxyl" refers to a-C (=o) OH group. The term "oxo" refers to an =o group.

The term "alkanoyl" refers to an alkyl group as defined herein attached to the remainder of the molecule through a-C (=o) group. Exemplary alkanoyl groups include, but are not limited to, acetyl, n-propionyl, and n-butyryl.

The term "alkanoyloxy" refers to an alkyl group as defined herein attached to the rest of the molecule through an-OC (=o) group. Exemplary alkanoyloxy groups include, but are not limited to, acetoxy, n-propionyloxy, and n-butyryloxy.

The term "alkoxycarbonyl" refers to an alkoxy group as defined herein (i.e., an alkyl ester group) attached to a C (=o) -alkyl group through an oxygen atom of the alkoxy group.

The terms "arylalkyl" and "cycloalkylalkyl" refer to an aryl or cycloalkyl group (as defined herein) attached to the remainder of the molecule through an alkyl group as defined herein. Exemplary arylalkyl groups include, but are not limited to, benzyl and phenethyl. Exemplary cycloalkylalkyl groups include, but are not limited to, cyclopropylmethyl, cyclobutylmethyl, cyclopentylethyl, and cyclohexylethyl.

With respect to any of the above groups containing one or more substituents, it is to be understood that such groups do not contain any substitution or pattern of substitution that is sterically impractical and/or synthetically infeasible. Furthermore, as further described herein, the compounds of the present disclosure may include all stereochemical isomers (and racemic mixtures) resulting from the substitution of these compounds.

As used herein, the term "amino acid" includes residues of natural amino acids in the D or L form (e.g., ala, arg, asn, asp, cys, glu, gln, gly, his, hyl, hyp, ile, leu, lys, met, phe, pro, ser, thr, trp, tyr and Val), as well as unnatural amino acids (e.g., phosphoserine, phosphothreonine, phosphotyrosine, hydroxyproline, γ -carboxyglutamic acid; hippuric acid, octahydroindole-2-carboxylic acid, statin, 1,2,3,4, -tetrahydroisoquinoline-3-carboxylic acid, penicillamine, ornithine, citrulline, α -methyl-alanine, p-benzoylphenylalanine, phenylglycine, propargylglycine, sarcosine, and t-butylglycine). The term also includes natural and unnatural amino acids with conventional amino protecting groups (e.g., acetyl or benzyloxycarbonyl), as well as amino acids that are substituted at the carboxy terminus (e.g., as (C) ₁ -C ₆ ) Alkyl, phenyl or benzyl esters or amides; or as alpha-methylbenzylamide) protected natural and unnatural amino acids. Other suitable amino and carboxyl protecting groups are known to those skilled in the art (see, e.g., T.W.Greene, protecting Groups In Organic Synthesis; wiley: new York,1981, and thereto)References cited therein). The amino acid may be linked to the remainder of the compound of formula I by the carboxy terminus, the amino terminus, or by any other convenient attachment point, for example by the sulphur of cysteine.

As used herein, the term "peptide" describes a sequence of at least about 2 and no more than about 25 amino acid and/or peptide residue. The peptide sequence may be linear or cyclic. For example, cyclic peptides may be prepared by or may be produced by disulfide bond formation between two cysteine residues in the sequence. The peptide may be linked to the remainder of the compound of formula I by the carboxy terminus, the amino terminus, or by any other convenient attachment point, for example by the sulphur of cysteine. In some cases, the peptides herein comprise 3 to 25, 5 to 21, or 10 to 25 amino acids. Peptide derivatives may be as described in U.S. patent No. 4,612,302;4,853,371; and 4,684,620. The peptide sequences specifically recited herein are written with the amino terminus to the left and the carboxy terminus to the right. As used herein, "protein" generally refers to a molecule comprising a sequence of at least about 25 amino acid and/or peptide residues, such as from about 25 to about 750, from about 50 to about 500, from about 100 to about 500, or from about 100 to about 1000.

As used herein, the term "free drug" refers to a biologically active drug molecule (e.g., one of formulas I-IV) that is not covalently attached to another moiety, such as a peptide or protein. Thus, free drug refers to a compound that exists immediately after cleavage from a conjugate (e.g., a drug-peptide or drug-protein conjugate as described herein). The release mechanism may be through a cleavable linker in the conjugate, or through intracellular transformation or metabolism of the conjugate. In some aspects, the free drug may be protonated and/or may be present as a charged moiety. Free drugs are pharmacologically active substances that are capable of exerting a specific biological effect. In some embodiments, the pharmacologically active substance is the sole parent drug. In some embodiments, the pharmacologically active substance is a parent drug bound to another molecule, for example in a conjugate.

Unless the context indicates or implies otherwise, the term "treatment" refers to therapeutic treatment and prophylactic measures to prevent recurrence, wherein the aim is to inhibit an undesired physiological change or disorder, such as the development or spread of cancer. For the purposes of this disclosure, beneficial or desired clinical results include, but are not limited to, alleviation of symptoms, diminishment of extent of disease, stabilized (e.g., non-worsening) state of disease, delay or slowing of disease progression, amelioration or palliation of the disease state, and remission (whether partial or total), whether detectable or undetectable. In some aspects, "treating" also includes extending the survival of the subject as compared to the expected survival without treatment.

In the case of cancer, the term "treatment" includes any or all of the following: inhibiting the growth of cancer cells or tumors; inhibit the replication of cancer cells, reduce overall tumor burden or reduce the number of cancer cells, and ameliorate one or more symptoms associated with the disease.

TLR modulating compounds

In some embodiments, provided herein are Toll-like receptor (TLR) modulatory compounds that can trigger a more desirable ratio of pro-inflammatory cytokines to anti-inflammatory cytokines. In some cases, compounds of the present disclosure can interact with and/or modulate TLR-7, TLR-8, or a combination of such receptors.

Structure of compound

In some cases, provided herein are compounds of formula I that are capable of modulating a TLR:

(I)

wherein:

fused ring a is selected from the group consisting of:

R ² is H, (C) ₁ -C ₆ ) Alkyl, (C) ₂ -C ₆ ) Alkenyl group (C) ₂ -C ₆ ) Alkynyl, (C) ₁ -C ₆ ) Alkanoyl, (C) ₁ -C ₆ ) Alkoxycarbonyl, (C) ₁ -C ₆ ) Alkanoyloxy, (C) ₃ -C ₆ ) Cycloalkyl, aryl, heteroaryl, heterocycle, NR ^g R ^h Or R is ^m R ⁿ NC (=o) -, any of which (C) ₁ -C ₆ ) Alkyl, (C) ₂ -C ₆ ) Alkenyl group (C) ₂ -C ₆ ) Alkynyl, (C) ₁ -C ₆ ) Alkanoyl, (C) ₁ -C ₆ ) Alkoxycarbonyl, (C) ₁ -C ₆ ) Alkanoyloxy, (C) ₃ -C ₆ ) Cycloalkyl, aryl, heteroaryl, and heterocycle are optionally substituted with one or more groups independently selected from the group consisting of: hydroxy, halo-SH, cyano, oxo, oxirane, (C) ₃ -C ₈ ) Cycloalkyl, aryl, heteroaryl, (C) ₁ -C ₆ ) Alkoxy, (C) ₁ -C ₆ ) Alkylthio and NR ^g R ^h ；

R ^a is H, (C) ₁ -C ₆ ) Alkyl, (C) ₂ -C ₆ ) Alkenyl group (C) ₂ -C ₆ ) Alkynyl, (C) ₁ -C ₆ ) Alkanoyl, (C) ₁ -C ₆ ) Alkoxycarbonyl, (C) ₃ -C ₆ ) Cycloalkyl, aryl or heteroaryl, any of which (C ₁ -C ₆ ) Alkyl, (C) ₂ -C ₆ ) Alkenyl group (C) ₂ -C ₆ ) Alkynyl, (C) ₁ -C ₆ ) Alkanoyl, (C) ₁ -C ₆ ) Alkoxycarbonyl, (C) ₃ -C ₆ ) Cycloalkyl, aryl, and heteroaryl are optionally substituted with one or more groups independently selected from the group consisting of: hydroxy, halo, -SH, cyano, oxirane, (C) ₃ -C ₈ ) Cycloalkyl, aryl, heteroaryl, (C) ₁ -C ₆ ) Alkoxy, (C) ₁ -C ₆ ) Alkylthio and NR ^g R ^h ；

R ^b Is H or X-Y;

R ^e is H, (C) ₁ -C ₆ ) Alkyl or R ^aa ；

R ^f Is H or (C) ₁ -C ₆ ) An alkyl group;

each R ^g And R is ^h Independently H, (C) ₁ -C ₆ ) Alkyl, (C) ₂ -C ₆ ) Alkenyl group (C) ₃ -C ₈ ) Cycloalkyl, (C) ₃ -C ₈ ) Cycloalkyl (C) ₁ -C ₆ ) Alkyl, aryl (C) ₁ -C ₆ ) An alkyl group; or together with the nitrogen to which they are attached form an aziridinyl, azetidinyl, morpholino, piperazino, pyriA fludioxo or piperidyl ring, said ring being optionally substituted by one or more (C ₁ -C ₆ ) Alkyl substitution;

x is a linking group; and is also provided with

Y is a peptide, protein or maleimide;

each R ^p And R is ^q Independently H or (C) ₁ -C ₆ ) An alkyl group; or together with the nitrogen to which they are attached form an aziridinyl group,Azetidinyl, morpholino, piperazino, pyrrolidino, or piperidinyl rings, which rings can optionally be substituted with one or more (C ₁ -C ₆ ) Alkyl substitution; and is also provided with

or a pharmaceutically acceptable salt thereof.

(I)

wherein:

fused ring a is selected from the group consisting of:

R ¹ is H, (C) ₁ -C ₆ ) Alkyl, (C) ₂ -C ₆ ) Alkenyl group (C) ₂ -C ₆ ) Alkynyl, (C) ₁ -C ₆ ) Alkanoyl, (C) ₁ -C ₆ ) Alkoxycarbonyl, (C) ₁ -C ₆ ) Alkanoyloxy, (C) ₃ -C ₆ ) Cycloalkyl, aryl, heteroaryl, heterocycle, NR ^g R ^h Or R is ^m R ⁿ NC (=o) -, any of which (C) ₁ -C ₆ ) Alkyl, (C) ₂ -C ₆ ) Alkenyl group (C) ₂ -C ₆ ) Alkynyl, (C) ₁ -C ₆ ) Alkanoyl, (C) ₁ -C ₆ ) Alkoxycarbonyl, (C) ₁ -C ₆ ) Alkanoyloxy, (C) ₃ -C ₆ ) Cycloalkyl, aryl, heteroaryl, and heterocycle are optionally substituted with one or more groups independently selected from the group consisting of: halo, cyano, oxo, oxirane, (C) ₃ -C ₈ ) Cycloalkyl, aryl optionally substituted by alkyl, aryl substituted by carboxyl, heteroaryl, (C) ₁ -C ₆ ) Alkoxy, (C) ₁ -C ₆ ) Alkylthio, OR ^z 、-N(H)S(O) ₂ R ^r 、R ^s C(＝O)O-、-S-R ^w 、-NR ^x R ^y 、(C ₁ -C ₆ ) Alkoxycarbonyl and carboxyl, said alkyl being substituted by NR ^u R ^v Substitution;

R ² is H, (C) ₁ -C ₆ ) Alkyl, (C) ₂ -C ₆ ) Alkenyl group (C) ₂ -C ₆ ) Alkynyl, (C) ₁ -C ₆ ) Alkanoyl, (C) ₁ -C ₆ ) Alkoxycarbonyl, (C) ₁ -C ₆ ) Alkanoyloxy, (C) ₃ -C ₆ ) Cycloalkyl, aryl, heteroaryl, heterocycle, NR ^g R ^h Or R is ^m R ⁿ NC (=o) -, any of which (C) ₁ -C ₆ ) Alkyl, (C) ₂ -C ₆ ) Alkenyl group (C) ₂ -C ₆ ) Alkynyl, (C) ₁ -C ₆ ) Alkanoyl, (C) ₁ -C ₆ ) Alkoxycarbonyl, (C) ₁ -C ₆ ) Alkanoyloxy, (C) ₃ -C ₆ ) Cycloalkyl, aryl, heteroaryl, and heterocycle are optionally substituted with one or more groups independently selected from the group consisting of: hydroxy, halo, -SH, cyano, oxo, oxirane, (C) ₃ -C ₈ ) Cycloalkyl, aryl, heteroaryl, (C) ₁ -C ₆ ) Alkoxy, (C) ₁ -C ₆ ) Alkylthio and NR ^g R ^h ；

R ⁴ Is R ^w S(O) ₂ NR ^f -；

R ^b Is H or X-Y;

each R ^c And R is ^d Independently H, (C) ₁ -C ₆ ) Alkyl, (C) ₂ -C ₆ ) Alkenyl group (C) ₃ -C ₈ ) Cycloalkyl, (C) ₃ -C ₈ ) Cycloalkyl (C) ₁ -C ₆ ) Alkyl, aryl (C) ₁ -C ₆ ) An alkyl group; or together with the nitrogen to which they are attached form an aziridinyl, azetidinyl, morpholino, piperazino, pyrrolidino or piperidinyl ring, said ring being optionally substituted with one or more (C ₁ -C ₆ ) Alkyl substitution;

R ^e is H, (C) ₁ -C ₆ ) Alkyl or R ^aa ；

R ^f Is H or (C) ₁ -C ₆ ) An alkyl group;

each R ^g And R is ^h Independently H, (C) ₁ -C ₆ ) Alkyl, (C) ₂ -C ₆ ) Alkenyl group (C) ₃ -C ₈ ) Cycloalkyl, (C) ₃ -C ₈ ) Cycloalkyl (C) ₁ -C ₆ ) Alkyl, aryl (C) ₁ -C ₆ ) An alkyl group; or together with the nitrogen to which they are attached form an aziridinyl group, Azetidinyl, morpholino, piperazino, pyrrolidino, or piperidinyl rings, which rings can optionally be substituted with one or more (C ₁ -C ₆ ) Alkyl substitution;

x is a linking group; and is also provided with

Y is a peptide, protein or maleimide;

each R ^p And R is ^q Independently H or (C) ₁ -C ₆ ) An alkyl group; or with themThe attached nitrogen groups together form an aziridinyl, azetidinyl, morpholino, piperazino, pyrrolidino or piperidinyl ring, which can optionally be substituted with one or more (C ₁ -C ₆ ) Alkyl substitution; and is also provided with

or a pharmaceutically acceptable salt thereof.

(I)

wherein:

fused ring a is selected from the group consisting of:

R ⁴ Is at the 2-position by R ^k -C(＝O)-、R ^k -O-C(＝O)-、R ^k -S(O) ₂ -O-、R ^c R ^d NC(＝O)-、R ^c R ^d NS(O) ₂ -、R ^e C(＝O)N(R ^e ) -or R ^e S(O) ₂ NR ^f -substituted 1-ethylene;

R ^b Is H or X-Y;

R ^e is H, (C) ₁ -C ₆ ) Alkyl or R ^aa ；

R ^f Is H or (C) ₁ -C ₆ ) An alkyl group;

each R ^g And R is ^h Independently H, (C) ₁ -C ₆ ) Alkyl, (C) ₂ -C ₆ ) Alkenyl group (C) ₃ -C ₈ ) Cycloalkyl, (C) ₃ -C ₈ ) Cycloalkyl (C) ₁ -C ₆ ) Alkyl, aryl (C) ₁ -C ₆ ) An alkyl group; or together with the nitrogen to which they are attached form an aziridinyl, azetidinyl, morpholino, piperazino, pyrrolidino or piperidinyl ring, said ring being optionally substituted with one or more (C ₁ -C ₆ ) Alkyl substitution;

x is a linking group; and is also provided with

Y is a peptide, protein or maleimide;

wherein rings B and C in formula I can optionally be one or more independently selected from halo, hydroxy, nitro, (C) at one or more carbons ₁ -C ₆ ) Alkyl, (C) ₁ -C ₆ ) Alkenyl group (C) ₁ -C ₆ ) Alkynyl, (C) ₁ -C ₆ ) Alkoxy, (C) ₁ -C ₆ ) Alkanoyl, (C) ₁ -C ₆ ) Alkanoyloxy radical、(C ₁ -C ₆ ) Alkoxycarbonyl, trifluoromethyl, trifluoromethoxy, cyano and NR ^p R ^q Further substitution of the groups of the group;

each R ^p And R is ^q Independently H or (C) ₁ -C ₆ ) An alkyl group; or together with the nitrogen to which they are attached form an aziridinyl, azetidinyl, morpholino, piperazino, pyrrolidino or piperidinyl ring, said ring being optionally substituted with one or more (C ₁ -C ₆ ) Alkyl substitution; and is also provided with

or a pharmaceutically acceptable salt thereof.

(I)

wherein:

the condensed ring A is:

R ^b Is H or X-Y;

x is a linking group; and is also provided with

Y is a peptide, protein or maleimide

(I)

wherein:

the condensed ring A is:

R ² is H, (C) ₁ -C ₆ ) Alkyl, (C) ₂ -C ₆ ) Alkenyl group (C) ₂ -C ₆ ) Alkynyl, (C) ₁ -C ₆ ) Alkanoyl, (C) ₁ -C ₆ ) Alkoxycarbonyl, (C) ₁ -C ₆ ) Alkanoyloxy, (C) ₃ -C ₆ ) Cycloalkyl, aryl, heteroaryl, heterocycle, NR ^g R ^h Or R is ^m R ⁿ NC (=o) -, any of which (C) ₁ -C ₆ ) Alkyl, (C) ₂ -C ₆ ) Alkenyl group (C) ₂ -C ₆ ) Alkynyl, (C) ₁ -C ₆ ) Alkanoyl, (C) ₁ -C ₆ ) Alkoxycarbonyl, (C) ₁ -C ₆ ) Alkanoyloxy, (C) ₃ -C ₆ ) Cycloalkyl, aryl, heteroaryl, and heterocycle are optionally substituted with one or more groups independently selected from the group consisting of: hydroxy, halo, -SH, cyano, oxo, oxirane, (C) ₃ -C ₈ ) Cycloalkyl group,Aryl, heteroaryl, (C) ₁ -C ₆ ) Alkoxy, (C) ₁ -C ₆ ) Alkylthio and NR ^g R ^h ；

R ^b Is H or X-Y;

each R ^g And R is ^h Independently H, (C) ₁ -C ₆ ) Alkyl, (C) ₂ -C ₆ ) Alkenyl group (C) ₃ -C ₈ ) Cycloalkyl, (C) ₃ -C ₈ ) Cycloalkyl (C) ₁ -C ₆ ) Alkyl, aryl (C) ₁ -C ₆ ) An alkyl group; or with themThe attached nitrogen groups together form an aziridinyl, azetidinyl, morpholino, piperazino, pyrrolidino or piperidinyl ring, which can optionally be substituted with one or more (C ₁ -C ₆ ) Alkyl substitution;

x is a linking group; and is also provided with

Y is a peptide, protein or maleimide.

(I)

wherein:

the condensed ring A is:

R ^b Is H or X-Y;

x is a linking group; and is also provided with

Y is a peptide, protein or maleimide.

(I)

wherein:

the condensed ring A is:

R ¹ is H, (C) ₁ -C ₆ ) Alkyl, (C) ₂ -C ₆ ) Alkenyl group (C) ₂ -C ₆ ) Alkynyl, (C) ₁ -C ₆ ) Alkanoyl group、(C ₁ -C ₆ ) Alkoxycarbonyl, (C) ₁ -C ₆ ) Alkanoyloxy, (C) ₃ -C ₆ ) Cycloalkyl, aryl, heteroaryl, heterocycle, NR ^g R ^h Or R is ^m R ⁿ NC (=o) -, any of which (C) ₁ -C ₆ ) Alkyl, (C) ₂ -C ₆ ) Alkenyl group (C) ₂ -C ₆ ) Alkynyl, (C) ₁ -C ₆ ) Alkanoyl, (C) ₁ -C ₆ ) Incubating carbonyl group (C) ₁ -C ₆ ) Alkanoyloxy, (C) ₃ -C ₆ ) Cycloalkyl, aryl, heteroaryl, and heterocycle are optionally substituted with one or more groups independently selected from the group consisting of: halo, cyano, oxo, oxirane, (C) ₃ -C ₈ ) Cycloalkyl, aryl optionally substituted by alkyl, aryl substituted by carboxyl, heteroaryl, (C) ₁ -C ₆ ) Alkoxy, (C) ₁ -C ₆ ) Alkylthio, OR ^z 、-N(H)S(O) ₂ R ^r 、R ^s C(＝O)O-、-S-R ^w 、-NR ^x R ^y 、(C ₁ -C ₆ ) Alkoxycarbonyl and carboxyl, said alkyl being substituted by NR ^u R ^v Substitution;

R ² Is H, (C) ₁ -C ₆ ) Alkyl, (C) ₂ -C ₆ ) Alkenyl group (C) ₂ -C ₆ ) Alkynyl, (C) ₁ -C ₆ ) Alkanoyl, (C) ₁ -C ₆ ) Alkoxycarbonyl, (C) ₁ -C ₆ ) Alkanoyloxy, (C) ₃ -C ₆ ) Cycloalkyl, aryl, heteroaryl, heterocycle, NR ^g R ^h Or R is ^m R ⁿ NC (=o) -, any of which (C) ₁ -C ₆ ) Alkyl, (C) ₂ -C ₆ ) Alkenyl group (C) ₂ -C ₆ ) Alkynyl, (C) ₁ -C ₆ ) Alkanoyl, (C) ₁ -C ₆ ) Alkoxycarbonyl, (C) ₁ -C ₆ ) Alkanoyloxy, (C) ₃ -C ₆ ) Cycloalkyl, aryl, heteroaryl, and heterocycle are optionally substituted with one or more groups independently selected from the group consisting of: hydroxy, halo, -SH, cyano, oxo, oxirane, (C) ₃ -C ₈ ) Cycloalkyl, aryl, heteroaryl, (C) ₁ -C ₆ ) Alkoxy, (C) ₁ -C ₆ ) Alkylthio and NR ^g R ^h The method comprises the steps of carrying out a first treatment on the surface of the And is also provided with

R ^b Is H or X-Y;

R ^e is H, (C) ₁ -C ₆ ) Alkyl or R ^aa ；

R ^f Is H or (C) ₁ -C ₆ ) An alkyl group;

each R ^g And R is ^h Independently H, (C) ₁ -C ₆ ) Alkyl, (C) ₂ -C ₆ ) Alkenyl group (C) ₃ -C ₈ ) Cycloalkyl, (C) ₃ -C ₈ ) Cycloalkyl (C) ₁ -C ₆ ) Alkyl, aryl (C) ₁ -C ₆ ) An alkyl group; or together with the nitrogen to which they are attached form an aziridinyl, azetidinyl, morpholino, piperazino, pyrrolidino or piperidinyl ring, which can optionally beOne or more (C) ₁ -C ₆ ) Alkyl substitution;

x is a linking group; and is also provided with

Y is a peptide, protein or maleimide;

each R ^p And R is ^q Independently H or (C) ₁ -C ₆ ) An alkyl group; or together with the nitrogen to which they are attached form aziridinyl, azetidinyl, morpholino, piperazino, pyrrolidineAnd or a piperidinyl ring, said ring being optionally substituted with one or more (C ₁ -C ₆ ) Alkyl substitution; and is also provided with

or a pharmaceutically acceptable salt thereof.

In some cases provided herein, the compound of formula I comprises a structure of formula III or IV:

or a pharmaceutically acceptable salt thereof,

wherein:

R ² is H, (C) ₁ -C ₆ ) Alkyl, (C) ₂ -C ₆ ) Alkenyl group (C) ₂ -C ₆ ) Alkynyl, (C) ₁ -C ₆ ) Alkanoyl, (C) ₁ -C ₆ ) Alkoxycarbonyl, (C) ₁ -C ₆ ) Alkanoyloxy, (C) ₃ -C ₆ ) Cycloalkyl, aryl, heteroaryl, heterocycle, NR ^g R ^h Or R is ^m R ⁿ NC (=o) -, any of which (C) ₁ -C ₆ ) Alkyl, (C) ₂ -C ₆ ) Alkenyl group (C) ₂ -C ₆ ) Alkynyl, (C) ₁ -C ₆ ) Alkyl (C)Acyl group (C) ₁ -C ₆ ) Alkoxycarbonyl, (C) ₁ -C ₆ ) Alkanoyloxy, (C) ₃ -C ₆ ) Cycloalkyl, aryl, heteroaryl, and heterocycle are optionally substituted with one or more groups independently selected from the group consisting of: hydroxy, halo, -SH, cyano, oxo, oxirane, (C) ₃ -C ₈ ) Cycloalkyl, aryl, heteroaryl, (C) ₁ -C ₆ ) Alkoxy, (C) ₁ -C ₆ ) Alkylthio and NR ^g R ^h ；

R ⁴ Is R ^k -C(＝O)-、R ^k -O-C (=o) -or R ^k -S(O) ₂ -O-；

R ^b Is H or X-Y;

each R ^g And R is ^h Independently H, (C) ₁ -C ₆ ) Alkyl, (C) ₂ -C ₆ ) Alkenyl group (C) ₃ -C ₈ ) Cycloalkyl, (C) ₃ -C ₈ ) Cycloalkyl (C) ₁ -C ₆ ) Alkyl, aryl (C) ₁ -C ₆ ) An alkyl group; or together with the nitrogen to which they are attached form an aziridinyl, azetidinyl, morpholino, piperazino, pyrrolidino or piperaquineA pyridyl ring, said ring being optionally substituted with one or more (C ₁ -C ₆ ) Alkyl substitution;

x is a linking group; and is also provided with

Y is a peptide, protein or maleimide.

(I)

wherein:

fused ring a is selected from the group consisting of:

R ^a is H, (C) ₁ -C ₆ ) Alkyl, (C) ₂ -C ₆ ) Alkenyl group (C) ₂ -C ₆ ) Alkynyl, (C) ₁ -C ₆ ) Alkanoyl, (C) ₁ -C ₆ ) Alkoxycarbonyl, (C) ₃ -C ₆ ) Cycloalkyl, aryl or heteroaryl, any of which (C ₁ -C ₆ ) Alkyl, (C) ₂ -C ₆ ) Alkenyl group (C) ₂ -C ₆ ) Alkynyl, (C) ₁ -C ₆ ) Alkanoyl, (C) ₁ -C ₆ ) Alkoxycarbonyl, (C) ₃ -C ₆ ) Cycloalkyl, aryl and heteroaryl are optionally independently selected from one or more ofSubstitution of groups from the group consisting of: hydroxy, halo, -SH, cyano, oxirane, (C) ₃ -C ₈ ) Cycloalkyl, aryl, heteroaryl, (C) ₁ -C ₆ ) Alkoxy, (C) ₁ -C ₆ ) Alkylthio and NR ^g R ^h ；

R ^b Is H or X-Y;

R ^e is H, (C) ₁ -C ₆ ) Alkyl or R ^aa ；

R ^f Is H or (C) ₁ -C ₆ ) An alkyl group;

R ^k is H, (C) ₁ -C ₆ ) Alkyl, (C) ₂ -C ₆ ) Alkenyl group (C) ₂ -C ₆ ) Alkynyl, (C) ₃ -C ₈ ) Cycloalkyl, trifluoromethyl, aryl or aryl (C) ₁ -C ₆ ) Alkyl groups, each of which (C ₁ -C ₆ ) The alkyl group being optionally substituted by one or more halo groups, (C) ₁ -C ₆ ) Alkanoyl compoundsOxy, (C) ₁ -C ₆ ) Alkoxy, (C) ₃ -C ₈ ) Cycloalkyl substitution;

x is a linking group; and is also provided with

Y is a peptide, protein or maleimide;

or a pharmaceutically acceptable salt thereof.

([)

wherein:

fused ring a is selected from the group consisting of:

R ¹ is H, (C) ₁ -C ₆ ) Alkyl, (C) ₂ -C ₆ ) Alkenyl group (C) ₂ -C ₆ ) Alkynyl, (C) ₁ -C ₆ ) Alkanoyl, (C) ₁ -C ₆ ) Alkoxycarbonyl, (C) ₁ -C ₆ ) Alkanoyloxy, (C) ₃ -C ₆ ) Cycloalkyl, heterocycle, NR ^g R ^h Or R is ^m R ⁿ NC (=o) -, any of which (C) ₁ -C ₆ ) Alkyl, (C) ₂ -C ₆ ) Alkenyl group (C) ₂ -C ₆ ) Alkynyl, (C) ₁ -C ₆ ) Alkanoyl, (C) ₁ -C ₆ ) Alkoxycarbonyl, (C) ₁ -C ₆ ) Alkanoyloxy, (C) ₃ -C ₆ ) Cycloalkyl and heterocycle are optionally substituted with one or more groups independently selected from the group consisting of: halo, cyano, oxo, oxirane, (C) ₃ -C ₈ ) Cycloalkyl, aryl optionally substituted by alkyl, aryl substituted by carboxyl, heteroaryl, (C) ₁ -C ₆ ) Alkoxy, (C) ₁ -C ₆ ) Alkylthio, OR ^z 、-N(H)S(O) ₂ R ^r 、R ^s C(＝O)O-、-S-R ^w 、-NR ^x R ^y 、(C ₁ -C ₆ ) Alkoxycarbonyl and carboxyl, said alkyl being substituted by NR ^u R ^v Substitution;

R ² is H, (C) ₁ -C ₆ ) Alkyl, (C) ₂ -C ₆ ) Alkenyl group (C) ₂ -C ₆ ) Alkynyl, (C) ₁ -C ₆ ) Alkanoyl, (C) ₁ -C ₆ ) Alkoxycarbonyl, (C) ₁ -C ₆ ) Alkanoyloxy, (C) ₃ -C ₆ ) Cycloalkyl, aryl, heteroaryl, heterocycle, NR ^g R ^h Or R is ^m R ⁿ NC (=o) -, any of which (C) ₁ -C ₆ ) Alkyl, (C) ₂ -C ₆ ) Alkenyl group (C) ₂ -C ₆ ) Alkynyl, (C) ₁ -C ₆ ) Alkanoyl, (C) ₁ -C ₆ ) Alkoxycarbonyl, (C) ₁ -C ₆ ) Alkanoyloxy, (C) ₃ -C ₆ ) Cycloalkyl, aryl, heteroaryl, and heterocycle are optionally substituted with one or more groups independently selected from the group consisting of: hydroxy, halo,-SH, cyano, oxo, oxirane, (C) ₃ -C ₈ ) Cycloalkyl, aryl, heteroaryl, (C) ₁ -C ₆ ) Alkoxy, (C) ₁ -C ₆ ) Alkylthio and NR ^g R ^h ；

R ⁴ Is R ^k -C(＝O)-、R ^k -O-C(＝O)-、R ^k -S(O) ₂ -O-、R ^c R ^d NC(＝O)-、R ^c R ^d NS(O) ₂ -、R ^e S(O) ₂ NR ^f -, or at the 2-position by R ^k -C(＝O)-、R ^k -O-C(＝O)-、R ^k -S(O) ₂ -O-、R ^c R ^d NC(＝O)-、R ^c R ^d NS(O) ₂ -、R ^e C(＝O)N(R ^e ) -or R ^e S(O) ₂ NR ^f -substituted 1-ethylene;

R ^a is H, (C) ₁ -C ₆ ) Alkyl, (C) ₂ -C ₆ ) Alkenyl group (C) ₂ -C ₆ ) Alkynyl, (C) ₁ -C ₆ ) Alkanoyl or (C) ₁ -C ₆ ) Alkoxycarbonyl, any of which (C ₁ -C ₆ ) Alkyl, (C) ₂ -C ₆ ) Alkenyl group (C) ₂ -C ₆ ) Alkynyl, (C) ₁ -C ₆ ) Alkanoyl (C) ₁ -C ₆ ) The alkoxycarbonyl group is optionally substituted with one or more groups independently selected from the group consisting of: hydroxy, halo, -SH, cyano, oxirane, (C) ₃ -C ₈ ) Cycloalkyl, aryl, heteroaryl, (C) ₁ -C ₆ ) Alkoxy, (C) ₁ -C ₆ ) Alkylthio and NR ^g R ^h ；

R ^b Is H or X-Y;

R ^e is H, (C) ₁ -C ₆ ) Alkyl or R ^aa ；

R ^f Is H or (C) ₁ -C ₆ ) An alkyl group;

x is a linking group; and is also provided with

Y is a peptide, protein or maleimide;

each R ^p And R is ^q Independently H or (C) ₁ -C ₆ ) An alkyl group; or together with the nitrogen to which they are attached form an aziridinyl, azetidinyl, morpholino, piperazino, pyrrolidino or piperidinyl ring, said ring being optionally substituted with one or more (C ₁ -C ₆ ) Alkyl substitution; and is combined withAnd is also provided with

or a pharmaceutically acceptable salt thereof.

In some embodiments, the compound of formula I comprises a structure of formula (Ia) or formula (Ib):

In some embodiments, the compound of formula I comprises a structure of formula (Ia):

in some embodiments, the compound of formula I comprises a structure of formula (Ib):

in some embodiments, fused ring a is selected from the group consisting of:

in some embodiments, fused ring a isIn some embodiments, fused ring A is +.>In some embodiments, fused ring A is +.>In some embodiments, fused ring A is +.>In some embodiments, fused ring A is +.>In some embodiments, fused ring A is +.>

In some embodiments, R ¹ Is H, (C) ₁ -C ₆ ) Alkyl, (C) ₂ -C ₆ ) Alkene (E)Radical (C) ₂ -C ₆ ) Alkynyl, (C) ₁ -C ₆ ) Alkanoyl, (C) ₁ -C ₆ ) Alkoxycarbonyl, (C) ₁ -C ₆ ) Alkanoyloxy, (C) ₃ -C ₆ ) Cycloalkyl, heterocycle, NR ^g R ^h Or R is ^m R ⁿ NC (=o) -, any of which (C) ₁ -C ₆ ) Alkyl, (C) ₂ -C ₆ ) Alkenyl group (C) ₂ -C ₆ ) Alkynyl, (C) ₁ -C ₆ ) Alkanoyl, (C) ₁ -C ₆ ) Alkoxycarbonyl, (C) ₁ -C ₆ ) Alkanoyloxy, (C) ₃ -C ₆ ) Cycloalkyl and heterocycle are optionally substituted with one or more groups independently selected from the group consisting of: halo, cyano, oxo, oxirane, (C) ₃ -C ₈ ) Cycloalkyl, aryl optionally substituted by alkyl, aryl substituted by carboxyl, heteroaryl, (C) ₁ -C ₆ ) Alkoxy, (C) ₁ -C ₆ ) Alkylthio, OR ^z 、-N(H)S(O) ₂ R ^r 、R ^s C(＝O)O-、-S-R ^w 、-NR ^x R ^y 、(C ₁ -C ₆ ) Alkoxycarbonyl and carboxyl, said alkyl being substituted by NR ^u R ^v And (3) substitution.

In some embodiments, R ¹ Is H, (C) ₁ -C ₆ ) Alkyl, (C) ₂ -C ₆ ) Alkenyl group (C) ₂ -C ₆ ) Alkynyl, (C) ₁ -C ₆ ) Alkanoyl, (C) ₁ -C ₆ ) Alkoxycarbonyl, (C) ₁ -C ₆ ) Alkanoyloxy, (C) ₃ -C ₆ ) Cycloalkyl, heterocycle, NR ^g R ^h Or R is ^m R ⁿ NC (=o) -, any of which (C) ₁ -C ₆ ) Alkyl, (C) ₂ -C ₆ ) Alkenyl group (C) ₂ -C ₆ ) Alkynyl, (C) ₁ -C ₆ ) Alkanoyl, (C) ₁ -C ₆ ) Alkoxycarbonyl, (C) ₁ -C ₆ ) Alkanoyloxy, (C) ₃ -C ₆ ) Cycloalkyl and heterocycle are optionally substituted with one or more groups independently selected from the group consisting of: halo, cyano, oxo, oxirane, (C) ₃ -C ₈ ) Cycloalkyl, aryl optionally substituted by alkyl, aryl substituted by carboxyl, heteroaryl, (C) ₁ -C ₆ ) Alkoxy, (C) ₁ -C ₆ ) Alkylthio, OR ^z 、-N(H)S(O) ₂ R ^r 、R ^s C(＝O)O-、-S-R ^w 、-NR ^x R ^y 、(C ₁ -C ₆ ) Alkoxycarbonyl and carboxyl, said alkyl being substituted by NR ^u R ^v And (3) substitution.

In some embodiments, R ¹ Is H, (C) ₁ -C ₆ ) Alkyl, (C) ₂ -C ₆ ) Alkenyl group (C) ₂ -C ₆ ) Alkynyl, (C) ₁ -C ₆ ) Alkanoyl, (C) ₁ -C ₆ ) Alkoxycarbonyl or-NR ^g R ^h Any of (C) ₁ -C ₆ ) Alkyl, (C) ₂ -C ₆ ) Alkenyl group (C) ₂ -C ₆ ) Alkynyl, (C) ₁ -C ₆ ) Alkanoyl (C) ₁ -C ₆ ) The alkoxycarbonyl group is optionally substituted with one or more groups independently selected from the group consisting of: halo, cyano, oxo, oxirane, (C) ₃ -C ₈ ) Cycloalkyl, aryl optionally substituted by alkyl, aryl substituted by carboxyl, heteroaryl, (C) ₁ -C ₆ ) Alkoxy, (C) ₁ -C ₆ ) Alkylthio, OR ^z 、-N(H)S(O) ₂ R ^r 、R ^s C(＝O)O-、-S-R ^w 、-NR ^x R ^y 、(C ₁ -C ₆ ) Alkoxycarbonyl and carboxyl, said alkyl being substituted by NR ^u R ^v And (3) substitution.

In some embodiments, R ¹ Is H, (C) ₁ -C ₆ ) Alkyl, (C) ₂ -C ₆ ) Alkenyl group (C) ₂ -C ₆ ) Alkynyl, (C) ₁ -C ₆ ) Alkanoyl, (C) ₁ -C ₆ ) Alkoxycarbonyl or-NR ^g R ^h Any of (C) ₁ -C ₆ ) Alkyl, (C) ₂ -C ₆ ) Alkenyl group (C) ₂ -C ₆ ) Alkynyl, (C) ₁ -C ₆ ) Alkanoyl (C) ₁ -C ₆ ) Alkoxycarbonyl optionally being substituted by oneOr a plurality of groups independently selected from halo, cyano, oxo, oxirane, (C) ₃ -C ₈ ) Cycloalkyl groups.

In some embodiments, R ¹ Is H, (C) ₁ -C ₆ ) Alkyl, (C) ₂ -C ₆ ) Alkenyl group (C) ₂ -C ₆ ) Alkynyl or-NR ^g R ^h Any of (C) ₁ -C ₆ ) Alkyl, (C) ₂ -C ₆ ) Alkenyl group sum (C) ₂ -C ₆ ) Alkynyl groups are optionally substituted with one or more groups independently selected from halo, cyano, oxo, oxirane, (C) ₃ -C ₈ ) Cycloalkyl groups.

In some embodiments, R ¹ Is H or optionally substituted with one or more groups independently selected from the group consisting of halo and cyano (C ₁ -C ₆ ) An alkyl group. R is R ¹ Is H or (C) ₁ -C ₆ ) An alkyl group.

In some embodiments, R ¹ Is H. In some embodiments, R ¹ Is not H.

In some embodiments, when R ² Is NR ^g R ^h When R is ¹ Is R ^c R ^d NS(O) ₂ -or R ^e S(O) ₂ NR ^f -. In some cases, when R ² Is covered by one or more (C ₁ -C ₆ ) Alkylthio substituted (C) ₁ -C ₆ ) In the case of alkyl radicals, R ¹ Is R ^c R ^d NS(O) ₂ -or R ^e S(O) ₂ NR ^f -。

In some embodiments, R ² Is H, (C) ₁ -C ₆ ) Alkyl, (C) ₂ -C ₆ ) Alkenyl group (C) ₂ -C ₆ ) Alkynyl, (C) ₁ -C ₆ ) Alkanoyl, (C) ₁ -C ₆ ) Alkoxycarbonyl or-NR ^g R ^h Any of (C) ₁ -C ₆ ) Alkyl, (C) ₂ -C ₆ ) Alkenyl group (C) ₂ -C ₆ ) Alkynyl, (C) ₁ -C ₆ ) Alkanoyl (C) ₁ -C ₆ ) Alkyl (C)The oxycarbonyl group is optionally substituted with one or more groups independently selected from the group consisting of: hydroxy, halo, -SH, cyano, oxo, oxirane, (C) ₃ -C ₈ ) Cycloalkyl, aryl, heteroaryl, (C) ₁ -C ₆ ) Alkoxy, (C) ₁ -C ₆ ) Alkylthio and NR ^g R ^h 。

In some embodiments, R ² Is H, (C) ₁ -C ₆ ) Alkyl, (C) ₂ -C ₆ ) Alkenyl group (C) ₂ -C ₆ ) Alkynyl, (C) ₁ -C ₆ ) Alkanoyl, (C) ₁ -C ₆ ) Alkoxycarbonyl or-NR ^g R ^h Any of (C) ₁ -C ₆ ) Alkyl, (C) ₂ -C ₆ ) Alkenyl group (C) ₂ -C ₆ ) Alkynyl, (C) ₁ -C ₆ ) Alkanoyl (C) ₁ -C ₆ ) Alkoxycarbonyl groups optionally being independently selected from halo, -SH, cyano, (C) ₃ -C ₈ ) Cycloalkyl and NR ^g R ^h The groups of the group are substituted.

In some embodiments, R ² Is H, (C) ₁ -C ₆ ) Alkyl, (C) ₂ -C ₆ ) Alkenyl or (C) ₂ -C ₆ ) Alkynyl, any of which (C ₁ -C ₆ ) Alkyl, (C) ₂ -C ₆ ) Alkenyl group sum (C) ₂ -C ₆ ) Alkynyl groups are optionally substituted with one or more groups independently selected from halo, -SH, cyano, (C) ₃ -C ₈ ) Cycloalkyl and NR ^g R ^h The groups of the group are substituted.

In some embodiments, R ² Is (C) ₁ -C ₆ ) Alkyl, (C) ₂ -C ₆ ) Alkenyl or (C) ₂ -C ₆ ) Alkynyl, any of which (C ₁ -C ₆ ) Alkyl, (C) ₂ -C ₆ ) Alkenyl group sum (C) ₂ -C ₆ ) Alkynyl groups are optionally substituted with one or more groups independently selected from halo, -SH, cyano, (C) ₃ -C ₈ ) Cycloalkyl and NR ^g R ^h The groups of the group are substituted.

In some embodimentsWherein R is ² Is H or is optionally substituted with one or more substituents independently selected from halo, -SH, cyano and NR ^g R ^h Group-substituted (C) ₁ -C ₆ ) An alkyl group.

In some embodiments, R ² Wherein R is ² Is H or (C) ₁ -C ₆ ) An alkyl group.

In some embodiments, R ² Is H. In some embodiments, R ² Is not H.

In some embodiments, R ³ Is H, halo, hydroxy, (C) ₁ -C ₆ ) Alkyl, (C) ₂ -C ₆ ) Alkenyl group (C) ₂ -C ₆ ) Alkynyl, (C) ₁ -C ₆ ) Alkanoyl, (C) ₁ -C ₆ ) Alkoxycarbonyl or NR ^g R ^h Any of (C) ₁ -C ₆ ) Alkyl, (C) ₂ -C ₆ ) Alkenyl group (C) ₂ -C ₆ ) Alkynyl, (C) ₁ -C ₆ ) Alkanoyl (C) ₁ -C ₆ ) The alkoxycarbonyl group is optionally substituted with one or more groups independently selected from the group consisting of: hydroxy, halo, -SH, cyano, oxo, oxirane, (C) ₃ -C ₈ ) Cycloalkyl, aryl, heteroaryl, (C) ₁ -C ₆ ) Alkoxy, (C) ₁ -C ₆ ) Alkylthio and NR ^g R ^h 。

In some embodiments, R ³ Is H, halo, hydroxy, (C) ₁ -C ₆ ) Alkyl, (C) ₂ -C ₆ ) Alkenyl group (C) ₂ -C ₆ ) Alkynyl, (C) ₁ -C ₆ ) Alkanoyl, (C) ₁ -C ₆ ) Alkoxycarbonyl or NR ^g R ^h Any of (C) ₁ -C ₆ ) Alkyl, (C) ₂ -C ₆ ) Alkenyl group (C) ₂ -C ₆ ) Alkynyl, (C) ₁ -C ₆ ) Alkanoyl (C) ₁ -C ₆ ) Alkoxycarbonyl groups optionally being independently selected from halo, -SH, cyano, (C) ₃ -C ₈ ) Cycloalkyl and NR ^g R ^h The groups of the group are substituted.

In some embodiments, R ³ Is H, (C) ₁ -C ₆ ) Alkyl, (C) ₂ -C ₆ ) Alkenyl group (C) ₂ -C ₆ ) Alkynyl or NR ^g R ^h Any of (C) ₁ -C ₆ ) Alkyl, (C) ₂ -C ₆ ) Alkenyl group sum (C) ₂ -C ₆ ) Alkynyl groups are optionally substituted with one or more groups independently selected from halo, -SH, cyano, (C) ₃ -C ₈ ) Cycloalkyl and NR ^g R ^h The groups of the group are substituted.

In some embodiments, R ³ Is H or is optionally substituted with one or more substituents independently selected from halo, -SH, cyano, (C) ₃ -C ₈ ) Cycloalkyl and NR ^g R ^h Group-substituted (C) ₁ -C ₆ ) An alkyl group.

In some embodiments, R ³ Is H or (C) ₁ -C ₆ ) An alkyl group.

In some embodiments, R ³ Is H. In some embodiments, R ³ Is not H.

In some embodiments, R ⁴ Is R ^k -C(＝O)-、R ^k -O-C(＝O)-、R ^k -S(O) ₂ -O-、R ^c R ^d NC(＝O)-、R ^c R ^d NS(O) ₂ -、R ^e S(O) ₂ NR ^f -, or at the 2-position by R ^k -C(＝O)-、R ^k -O-C(＝O)-、R ^k -S(O) ₂ -O-、R ^c R ^d NC(＝O)-、R ^c R ^d NS(O) ₂ -、R ^e C(＝O)N(R ^e ) -or R ^e S(O) ₂ NR ^f -substituted 1-ethylene.

In some embodiments, R ⁴ Is at the 2-position by R ^k -C(＝O)-、R ^k -O-C(＝O)-、R ^k -S(O) ₂ -O-、R ^c R ^d NC(＝O)-、R ^c R ^d NS(O) ₂ -、R ^e C(＝O)N(R ^e ) -or R ^e S(O) ₂ NR ^f -substituted 1-ethylene.

In one placeIn some embodiments, R ⁴ Is R ^k -C(＝O)-、R ^k -O-C(＝O)-、R ^k -S(O) ₂ -O-、R ^c R ^d NS(O) ₂ -、R ^e S(O) ₂ NR ^f -, or at the 2-position by R ^k -C(＝O)-、R ^k -O-C(＝O)-、R ^k -S(O) ₂ -O-、R ^c R ^d NS(O) ₂ -or R ^e S(O) ₂ NR ^f -substituted 1-ethylene.

In some embodiments, R ⁴ Is R ^k -C(＝O)-、R ^k -O-C (=o) -, or at the 2-position by R ^k -C(＝O)-、R ^k -O-C (=o) -substituted 1-ethylene.

In some embodiments, R ⁴ Is R ^k -S(O) ₂ -O-、R ^c R ^d NS(O) ₂ -、R ^e S(O) ₂ NR ^f -, or at the 2-position by R ^k -S(O) ₂ -O-、R ^c R ^d NS(O) ₂ -or R ^e S(O) ₂ NR ^f -substituted 1-ethylene.

In some embodiments, R ⁴ Is R ^k -C (=o) -or R ^k -O-C(＝O)-。

In some cases, provided herein are compounds of formula I, wherein R ⁴ Is R ^k -C(＝O)-、R ^k -O-C(＝O)-、R ^k -S(O) ₂ -O-、R ^c R ^d NC(＝O)-、R ^c R ^d NS(O) ₂ -、R ^e C(＝O)N(R ^e ) -or R ^e S(O) ₂ NR ^f . In such cases, R ⁴ May be R ^e S(O) ₂ NR ^f -and R ^e May be R ^w . In other cases, R ⁴ Is at the 2-position by R ^k -C(＝O)-、R ^k -O-C(＝O)-、R ^k -S(O) ₂ -O-、R ^c R ^d NC(＝O)-、R ^c R ^d NS(O) ₂ -、R ^e C(＝O)N(R ^e ) -or R ^e S(O) ₂ NR ^f -substituted 1-ethylene. The compounds of formula I herein may also be those wherein R ^b Is HA compound. In other cases, R ^b Is X-Y. In such cases, X may be (C ₁ -C ₆ ) Alkyl, (C) ₂ -C ₆ ) Alkenyl or (C) ₁ -C ₆ ) Alkynyl, said (C ₁ -C ₆ ) Alkyl, (C) ₂ -C ₆ ) Alkenyl or (C) ₁ -C ₆ ) Alkynyl groups are optionally substituted with oxo groups.

In some embodiments, R ^a Is H, (C) ₁ -C ₆ ) Alkyl, (C) ₂ -C ₆ ) Alkenyl group (C) ₂ -C ₆ ) Alkynyl, (C) ₁ -C ₆ ) Alkanoyl or (C) ₁ -C ₆ ) Alkoxycarbonyl, any of which (C ₁ -C ₆ ) Alkyl, (C) ₂ -C ₆ ) Alkenyl group (C) ₂ -C ₆ ) Alkynyl, (C) ₁ -C ₆ ) Alkanoyl (C) ₁ -C ₆ ) The alkoxycarbonyl group is optionally substituted with one or more groups independently selected from the group consisting of: hydroxy, halo, -SH, cyano, oxirane, (C) ₃ -C ₈ ) Cycloalkyl, aryl, heteroaryl, (C) ₁ -C ₆ ) Alkoxy, (C) ₁ -C ₆ ) Alkylthio and NR ^g R ^h 。

In some embodiments, R ^a Is H, (C) ₁ -C ₆ ) Alkyl, (C) ₂ -C ₆ ) Alkenyl or (C) ₂ -C ₆ ) Alkynyl, any of which (C ₁ -C ₆ ) Alkyl, (C) ₂ -C ₆ ) Alkenyl or (C) ₂ -C ₆ ) Alkynyl groups are optionally substituted with one or more groups independently selected from the group consisting of: halo, -SH, cyano, oxirane, (C) ₃ -C ₈ ) Cycloalkyl, (C) ₁ -C ₆ ) Alkoxy, (C) ₁ -C ₆ ) Alkylthio and NR ^g R ^h 。

In some embodiments, R ^a Is H, (C) ₁ -C ₆ ) Alkyl, (C) ₂ -C ₆ ) Alkenyl or (C) ₂ -C ₆ ) Alkynyl, any of which (C ₁ -C ₆ ) Alkyl, (C) ₂ -C ₆ ) Alkenyl or (C) ₂ -C ₆ ) Alkynyl is optionally substituted with one or more groups independently selected from halo, -SH, cyano and NR ^g R ^h The groups of the group are substituted.

In some embodiments, R ^a Is H or is optionally substituted with one or more substituents independently selected from halo, -SH, cyano and NR ^g R ^h Group-substituted (C) ₁ -C ₆ ) An alkyl group.

In some embodiments, R ^a Is H or (C) ₁ -C ₆ ) An alkyl group.

In some embodiments, R ^a Is H. In some embodiments, R ^a Is not H.

In some embodiments, provided herein are compounds of formula I, wherein

Ring a is selected from the group consisting of rings i, j, k, l, u and w;

R ¹ 、R ² and R is ³ May independently be H or optionally substituted (C ₁ -C ₆ ) An alkyl group;

R ⁴ is R ^k -C(＝O)-、R ^k -O-C (=o) -or R ^k -S(O) ₂ -O-；

x is a linking group; and is also provided with

Y is a peptide, protein or maleimide.

In some embodiments, provided herein are compounds of formula I, wherein ring a is selected from the group consisting of:

wherein R is ² Is H, (C) ₁ -C ₆ ) Alkyl, (C) ₂ -C ₆ ) Alkenyl group (C) ₂ -C ₆ ) Alkynyl, (C) ₁ -C ₆ ) Alkanoyl, (C) ₁ -C ₆ ) Alkoxycarbonyl, (C) ₁ -C ₆ ) Alkanoyloxy, (C) ₃ -C ₆ ) Cycloalkyl, aryl, heteroaryl, heterocycle, NR ^g R ^h Or R is ^m R ⁿ NC (=o) -, any of which (C) ₁ -C ₆ ) Alkyl, (C) ₂ -C ₆ ) Alkenyl group (C) ₂ -C ₆ ) Alkynyl, (C) ₁ -C ₆ ) Alkanoyl, (C) ₁ -C ₆ ) Alkoxycarbonyl, (C) ₁ -C ₆ ) Alkanoyloxy, (C) ₃ -C ₆ ) Cycloalkyl, aryl, heteroaryl, and heterocycle are optionally substituted with one or more groups independently selected from the group consisting of: hydroxy, halo, -SH, cyano, oxo, oxirane, (C) ₃ -C ₈ ) Cycloalkyl, aryl, heteroaryl, (C) ₁ -C ₆ ) Alkoxy, (C) ₁ -C ₆ ) Alkylthio and NR ^g R ^h . In some cases, R ² Is H or optionally substituted (C) ₁ -C ₆ ) An alkyl group. In some cases, ring a is ring i. In some cases, ring a is ring j. In some cases, ring a is ring k. In some cases, ring a is ring i.

In the case where ring a is ring j, the compound may have formula II:

(II)

or a pharmaceutically acceptable salt thereof,

wherein:

R ² is H, (C) ₁ -C ₆ ) Alkyl, (C) ₂ -C ₆ ) Alkenyl group (C) ₂ -C ₆ ) Alkynyl, (C) ₁ -C ₆ ) Alkanoyl, (-)C ₁ -C ₆ ) Alkoxycarbonyl, (C) ₁ -C ₆ ) Alkanoyloxy, (C) ₃ -C ₆ ) Cycloalkyl, aryl, heteroaryl, heterocycle, NR ^g R ^h Or R is ^m R ⁿ NC (=o) -, any of which (C) ₁ -C ₆ ) Alkyl, (C) ₂ -C ₆ ) Alkenyl group (C) ₂ -C ₆ ) Alkynyl, (C) ₁ -C ₆ ) Alkanoyl, (C) ₁ -C ₆ ) Alkoxycarbonyl, (C) ₁ -C ₆ ) Alkanoyloxy, (C) ₃ -C ₆ ) Cycloalkyl, aryl, heteroaryl, and heterocycle are optionally substituted with one or more groups independently selected from the group consisting of: hydroxy, halo, -SH, cyano, oxo, oxirane, (C) ₃ -C ₈ ) Cycloalkyl, aryl, heteroaryl, (C) ₁ -C ₆ ) Alkoxy, (C) ₁ -C ₆ ) Alkylthio and NR ^g R ^h ；

R ⁴ Is R ^k -C(＝O)-、R ^k -O-C (=o) -or R ^k -S(O) ₂ -O-；

R ^b Is H or X-Y;

x is a linking group; and is also provided with

Y is a peptide, protein or maleimide.

In some embodiments of formula II:

(II)

or a pharmaceutically acceptable salt thereof,

wherein:

R ⁴ Is R ^k -C (=o) -or R ^k -O-C(＝O)-；

R ^a Is H or optionally substituted (C) ₁ -C ₆ ) An alkyl group;

R ^b is H or X-Y;

x is a linking group; and is also provided with

Y is a peptide, protein or maleimide.

In some embodiments, provided herein are compounds of formula II, wherein:

R ² is H or optionally substituted (C) ₁ -C ₆ ) An alkyl group;

R ⁴ is R ^k -C (=o) -or R ^k -O-C(＝O)-；

R ^a Is H or optionally substituted (C) ₁ -C ₆ ) An alkyl group;

R ^b is H or X-Y;

R ^k is H or optionally substituted (C) ₁ -C ₆ ) An alkyl group;

x is a linking group; and is also provided with

Y is an antigen or maleimide.

In some embodiments and referring to formula II, R ² Is optionally substituted (C) ₁ -C ₆ ) An alkyl group; r is R ^k -O-C (=o) -, wherein R ^k Is optionally substituted (C) ₁ -C ₆ ) An alkyl group; r is R ^a Is H; and R is ^b Is H or X-Y, wherein X is a linking group and Y is an antigen or maleimide.

Thus, in some embodiments, provided herein are compounds of formula I or II having the following structure of compound 1:

in some embodiments, provided herein are compounds of formula I, wherein ring a is:

wherein the method comprises the steps of

R ² is H, (C) ₁ -C ₆ ) Alkyl, (C) ₂ -C ₆ ) Alkenyl group (C) ₂ -C ₆ ) Alkynyl, (C) ₁ -C ₆ ) Alkanoyl, (C) ₁ -C ₆ ) Alkoxycarbonyl, (C) ₁ -C ₆ ) Alkanoyloxy, (C) ₃ -C ₆ ) Cycloalkyl, aryl, heteroaryl, heterocycle, NR ^g R ^h Or R is ^m R ⁿ NC (=o) -, any of which (C) ₁ -C ₆ ) Alkyl, (C) ₂ -C ₆ ) Alkenyl group (C) ₂ -C ₆ ) Alkynyl, (C) ₁ -C ₆ ) Alkanoyl, (C) ₁ -C ₆ ) Alkoxycarbonyl, (C) ₁ -C ₆ ) Alkanoyloxy, (C) ₃ -C ₆ ) Cycloalkyl, aryl, heteroaryl, and heterocycle are optionally substituted with one or more groups independently selected from the group consisting of: hydroxy, halo, -SH, cyano, oxo, oxirane, (C) ₃ -C ₈ ) Cycloalkyl, aryl, heteroaryl, (C) ₁ -C ₆ ) Alkyl (C)Oxy, (C) ₁ -C ₆ ) Alkylthio and NR ^g R ^h ；

R ³ Is H, halo, hydroxy, (C) ₁ -C ₆ ) Alkyl, (C) ₂ -C ₆ ) Alkenyl group (C) ₂ -C ₆ ) Alkynyl, (C) ₁ -C ₆ ) Alkanoyl, (C) ₁ -C ₆ ) Alkoxycarbonyl, (C) ₁ -C ₆ ) Alkanoyloxy, (C) ₃ -C ₆ ) Cycloalkyl, aryl, heteroaryl, heterocycle, NR ^g R ^h Or R is ^m R ⁿ NC (=o) -, any of which (C) ₁ -C ₆ ) Alkyl, (C) ₂ -C ₆ ) Alkenyl group (C) ₂ -C ₆ ) Alkynyl, (C) ₁ -C ₆ ) Alkanoyl, (C) ₁ -C ₆ ) Alkoxycarbonyl, (C) ₁ -C ₆ ) Alkanoyloxy, (C) ₃ -C ₆ ) Cycloalkyl, aryl, heteroaryl, and heterocycle are optionally substituted with one or more groups independently selected from the group consisting of: hydroxy, halo, -SH, cyano, oxo, oxirane, (C) ₃ -C ₈ ) Cycloalkyl, aryl, heteroaryl, (C) ₁ -C ₆ ) Alkoxy, (C) ₁ -C ₆ ) Alkylthio and NR ^g R ^h As described herein for formula I.

Thus, in the same aspect, provided herein are compounds of formula III and formula IV, respectively:

wherein,

R ¹ 、R ² and R is ³ May independently be H or may optionally be covered (C ₁ -C ₃ ) Alkyl, substituted or unsubstituted (C ₃ -C ₈ ) Cycloalkyl or substituted or unsubstituted aryl substituted (C ₁ -C ₆ ) An alkyl group;

R ⁴ is R ^k -C(＝O)-、R ^k -O-C (=o) -or R ^k -S(O) ₂ -O-；

R ^a is H or optionally substituted (C) ₁ -C ₆ ) An alkyl group;

R ^b is H or X-Y;

x is a linking group; and is also provided with

Y is an antigen or maleimide.

Thus, in some embodiments, the compounds herein are selected from the group consisting of:

in some embodiments, a compound of the present disclosure, e.g., a compound of any one of formulas I-IV, is capable of binding to a toll-like receptor (TLR). In some embodiments, binding of such compounds to a TLR may exhibit agonist action on the TLR. In some embodiments, binding of a compound to a TLR can exert an immunostimulatory effect.

Synthesis of Compounds

The process for preparing the compounds of formulae I-IV is provided as a further embodiment of the present disclosure and is illustrated by the following procedure, wherein the meaning of the general groups is given above unless otherwise defined. Certain compounds of formulas I-IV are useful as intermediates for preparing other compounds of formulas I-IV. Pharmaceutically acceptable salts of the compounds of formulas I-IV may be used as intermediates for isolating or purifying the compounds of formulas I-IV, in cases where the compounds are sufficiently basic or acidic.

Conventional synthetic methods for tricyclic A-B-C heterocycles as shown in formulas I-IV may be based on substituted quinoline backbones. However, such routes may be limited in their ability to generate multifunctional patterns on tricyclic a-B-C heterocycles due to the harsh reaction conditions that may be required. In addition, as further shown below, conventional synthetic methods may not yield 5N-oxide intermediates in the presence of electron withdrawing groups.

Thus, in some embodiments, provided herein are improved synthetic routes to such tricyclic a-B-C heterocycles (such as thiazolines) as shown in formulas I-IV, which may allow for the installation of various substituents (e.g., R ⁴ )。

In various aspects, the substituted quinoline compounds of the present disclosure, including those of formulas I-IV, can be produced according to scheme 1 below:

wherein:

z is S, O or-NR ^f ；

R ^e is H, (C) ₁ -C ₆ ) Alkyl or R ^aa ；

R ^f Is H or (C) ₁ -C ₆ ) An alkyl group;

each R ^m And R is ⁿ Independently H, (C) ₁ -C ₆ ) Alkyl, (C) ₂ -C ₆ ) Alkenyl group (C) ₃ -C ₈ ) Cycloalkyl, (C) ₃ -C ₈ ) Cycloalkyl (C) ₁ -C ₆ ) Alkyl, aryl (C) ₁ -C ₆ ) An alkyl group; or together with the nitrogen to which they are attached form an aziridinyl, azetidinyl, morpholino, piperazino, pyrrolidino or piperidinyl ring, said ring being optionally substituted with one or more (C ₁ -C ₆ ) Alkyl substitution; and is also provided with

R ^aa Is aryl optionally substituted with one or more groups independently selected from the group consisting of: hydroxy, halogen, nitro, cyano, (C) ₁ -C ₆ ) Alkoxy and optionally substituted with one or more halo (C ₁ -C ₆ ) An alkyl group.

Scheme 2 below shows a variant of scheme 1 above, including substitution of alternative compounds:

wherein:

w is N or CH;

z is S, O or-NR ^f ；

R ² Is H, (C) ₁ -C ₆ ) Alkyl, (C) ₂ -C ₆ ) Alkenyl group (C) ₂ -C ₆ ) Alkynyl, (C) ₁ -C ₆ ) Alkanoyl, (C) ₁ -C ₆ ) Alkoxycarbonyl, (C) ₁ -C ₆ ) Alkanoyloxy compoundsRadical (C) ₃ -C ₆ ) Cycloalkyl, aryl, heteroaryl, heterocycle, NR ^g R ^h Or R is ^m R ⁿ NC (=o) -, any of which (C) ₁ -C ₆ ) Alkyl, (C) ₂ -C ₆ ) Alkenyl group (C) ₂ -C ₆ ) Alkynyl, (C) ₁ -C ₆ ) Alkanoyl, (C) ₁ -C ₆ ) Alkoxycarbonyl, (C) ₁ -C ₆ ) Alkanoyloxy, (C) ₃ -C ₆ ) Cycloalkyl, aryl, heteroaryl, and heterocycle are optionally substituted with one or more groups independently selected from the group consisting of: hydroxy, halo, -SH, cyano, oxo, oxirane, (C) ₃ -C ₈ ) Cycloalkyl, aryl, heteroaryl, (C) ₁ -C ₆ ) Alkoxy, (C) ₁ -C ₆ ) Alkylthio and NR ^g R ^h ；

R ⁴ Is R ^k -C(＝O)-、R ^k -O-C(＝O)-、R ^k -S(O) ₂ -O-、R ^c R ^d NC(＝O)-、R ^c R ^d NS(O) ₂ -、R ^e C(＝O)N(R ^e )-、R ^e S(O) ₂ NR ^f -, or at the 2-position by R ^k -C(＝O)-、R ^k -O-C(＝O)-、R ^k -S(O) ₂ -O-、R ^c R ^d NC(＝O)-、R ^c R ^d NS(O) ₂ -、R ^e C(＝O)N(R ^e ) -or R ^e S(O) ₂ NR ^f -substituted 1-ethylene.

R ^e is H, (C) ₁ -C ₆ ) Alkyl orR ^aa ；

R ^f Is H or (C) ₁ -C ₆ ) An alkyl group;

In some embodiments and according to reaction schemes 1 and 2 above, R ² Is (C) ₁ -C ₆ ) An alkyl group.

In some embodiments and according to reaction schemes 1 and 2 above:

R ⁴ is aryl, (C) ₂ -C ₆ ) Alkynyl, vinyl, cyano, R ^k -C(＝O)-、R ^m R ⁿ NC (=o) -or R ^k -O-C(＝O)-；

R ^k Is H, (C) ₁ -C ₆ ) Alkyl, (C) ₂ -C ₆ ) Alkenyl group (C) ₂ -C ₆ ) Alkynyl, (C) ₃ -C ₈ ) Cycloalkyl, trifluoromethyl, aryl or aryl (C) ₁ -C ₆ ) Alkyl groups, each of which (C ₁ -C ₆ ) The alkyl group being optionally substituted by one or more halo groups, (C) ₁ -C ₆ ) Alkanoyloxy, (C) ₁ -C ₆ ) Alkoxy, (C) ₃ -C ₈ ) Cycloalkyl substitution; and is also provided with

Each R ^m And R is ⁿ Independently H, (C) ₁ -C ₆ ) Alkyl, (C) ₂ -C ₆ ) Alkenyl group (C) ₃ -C ₈ ) Cycloalkyl, (C) ₃ -C ₈ ) Cycloalkyl (C) ₁ -C ₆ ) Alkyl, aryl (C) ₁ -C ₆ ) An alkyl group; or together with the nitrogen to which they are attached form an aziridinyl, azetidinyl, morpholino, piperazino, pyrrolidino or piperidinyl ring, said ring being optionally substituted with one or more (C ₁ -C ₆ ) Alkyl substitution.

Alternatively, as shown in scheme 3 below, 7-bromo-3-nitroquinolin-4 (1H) one in scheme 1 above may be synthesized;

in some embodiments, provided herein is a procedure that may begin with the installation of a bromo group using a modified Niman Style reaction (Niementowski reaction)To thiazoloquinoline to give the desired 3-amino-4-hydroxy-7-bromoquinoline. Such 7-bromo-thiazoloquinolines can then be cyclized by first forming a primary amide at the 3-amino position followed by dehydration of the pyridine containing phosphorus pentasulfide in use. Oxidation of the N5 position with 3-chloroperbenzoic acid-containing methylene chloride in the presence of C7-bromo group can then be carried out in good yields (e.g.,>70%) to obtain N5-oxide. In such cases, no or only a very small amount of N3 oxidation products may be present. The nucleophilic dipolar benzoyl isocyanate can then be used to install 4-amino as a benzoylamino substitute by a 1, 3-dipolar cycloaddition reaction and release carbon dioxide as a by-product. The final product can then be obtained in a two-step process using Pd ₂ (dba) ₃ And a catalyst (e.g., XPhos) to form a C7-phenyl ester, followed by acid catalyzed hydrolysis and transesterification to give the target thiazoline compound as shown.

In some embodiments, provided herein are further methods of synthesis of various substituted quinolines capable of interacting with one or more TLRs (such as TLR-7 and/or TLR-8). In some aspects, provided herein are synthetic routes to substituted pyrroloquinolines, e.g., as shown in scheme 4:

wherein:

R ^e is H, (C) ₁ -C ₆ ) Alkyl or R ^aa ；

R ^f Is H or (C) ₁ -C ₆ ) An alkyl group;

each R ^u And R is ^v Independently H or (C) ₁ -C ₆ ) Alkyl (C)A base; or together with the nitrogen to which they are attached form an aziridinyl, azetidinyl, morpholino, piperazino, pyrrolidino or piperidinyl ring, said ring being optionally substituted with one or more (C ₁ -C ₆ ) Alkyl substitution;

other embodiments of the present disclosure provide synthetic routes to substituted furanquinolines, for example, as shown in scheme 5:

wherein:

R ^e is H, (C) ₁ -C ₆ ) Alkyl or R ^aa ；

R ^f Is H or (C) ₁ -C ₆ ) An alkyl group;

In some embodiments, substituents R of schemes 1-5 ¹ 、R ² And R is ⁴ Independently selected from H, (C) ₁ -C ₆ ) Alkyl, aryl, heteroaryl and (C) ₃ -C ₈ ) Cycloalkyl groups, all of which (except H) may optionally be substituted with (C) ₁ -C ₃ ) Alkyl, substituted or unsubstituted (C ₃ -C ₈ ) Cycloalkyl or substituted or unsubstituted aryl. In some embodiments, schemes 1-5 can be used to synthesize compounds of formulas I-IV, such as compounds 1-26 as intermediates or end products.

III. Joint

In some embodiments, provided herein are molecules that can comprise a compound of any one of formulas I-IV and a linker. In such cases, for example, the linking group X of formula I herein may link the remainder of the compounds of formulas I-IV to another molecule Y. Such other molecules Y may comprise or consist of small molecules, peptides, proteins, nucleic acids or combinations thereof. In some cases, X-Y may comprise a maleimide moiety coupled to antigen Y. In various aspects, the nature of the linking group X may not be critical, so long as the resulting antigen conjugate retains the biological properties of the unconjugated antigen.

In some embodiments, linker X may have a molecular weight of about 20 daltons to about 20,000 daltons, about 20 daltons to about 5,000 daltons, about 20 daltons to about 1,000 daltons, or about 20 daltons to about 200 daltons. In some cases, X may have a length of about 5 angstroms to about 60 angstroms. In some embodiments, linker X separates the antigen from the remainder of the compounds of formulas I-IV by a length of about 5 angstroms to about 40 angstroms (inclusive).

The linker moiety X of the present disclosure may comprise or consist of a divalent, branched or unbranched, saturated or unsaturated hydrocarbon chain having from about 2 to about 25 carbon atoms, wherein one or more (e.g., 1, 2, 3, or 4) carbon atoms are optionally substituted with (-O-), and wherein the chain is optionally substituted on carbon with one or more (e.g., 1, 2, 3, or 4) substituents selected from the group consisting of: (C) ₁ -C ₆ ) Alkoxy, (C) ₃ -C ₆ ) Cycloalkyl, (C) ₁ -C ₆ ) Alkanoyl, (C) ₁ -C ₆ ) Alkanoyloxy, (C) ₁ -C ₆ ) Alkoxycarbonyl, (C) ₁ -C ₆ ) Alkylthio, azido, cyano, nitro, halo, hydroxy, oxo (=o), carboxyl, aryl, aryloxy, heteroaryl and heteroaryloxy. In another embodiment, the linker X comprises or consists of a polyethyleneoxy chain. Such a polyethyleneoxy chain may comprise or consist of 2, 3, 4, 5, 6, 7, 8, 9 or 10 repeating ethyleneoxy units.

In some cases, the linker moiety X may be a divalent group formed by a protein. In some cases, the linker moiety X may be a divalent group formed from a peptide. In some cases, the linker moiety X may be a divalent group formed from an amino acid.

In some embodiments, linker moiety X may comprise or consist of a molecule selected from the group consisting of:

wherein n is 2, 3, 4, 5 or 6, and

wherein R is ^bb Is an amino acid side chain.

and wherein n is 2, 3, 4, 5 or 6.

conjugates of

The present disclosure further provides conjugates comprising one or more compounds of any of formulas I-IV coupled to another molecule Y.

In some cases, such one or more compounds of any of formulas I-IV may be coupled to another molecule Y through a linker, as described herein. In some cases, another molecule Y may be a biological molecule, such as a peptide or protein (e.g., an antibody or antigen binding fragment thereof). Such conjugates may be molecules of formula V, which are identical to formula I, wherein R ^b Is X-Y, and X is a linking group and Y is a peptide, protein or maleimide.

In some cases, the compounds of formulas I-IV herein are coupled to a biomolecule Y, such as a peptide or protein. In some cases, the biomolecule Y is an antibody. In other cases, the biomolecule Y is an antigen. Accordingly, provided herein are conjugates comprising one or more compounds of any of formulas I-IV coupled to an antigen Y through a linker X as described herein.

V. pharmaceutical composition

Provided herein are pharmaceutical compositions comprising one or more compounds of formulas I-IV and/or one or more conjugates according to the present disclosure. In some cases, it may be appropriate to administer the compound of formula I in the form of a pharmaceutically acceptable acid or base salt. Examples of pharmaceutically acceptable salts are organic acid addition salts formed with acids forming physiologically acceptable anions, such as tosylate, mesylate, acetate, citrate, malonate, tartrate, succinate, benzoate, ascorbate, alpha-ketoglutarate and alpha-glycerophosphate. Suitable inorganic salts may also be formed, including hydrochloride, sulfate, nitrate, bicarbonate, and carbonate.

Pharmaceutically acceptable salts can be obtained using standard procedures well known in the art, for example by reacting a sufficiently basic compound such as an amine with a suitable acid that provides a physiologically acceptable anion. Alkali metal (e.g., sodium, potassium, or lithium) or alkaline earth metal (e.g., calcium) salts of carboxylic acids may also be prepared.

The compounds of formula I may be formulated into pharmaceutical compositions and administered to a mammalian host in various forms suitable for the chosen route of administration, such as human patients, for example orally or parenterally, by intravenous, intramuscular, topical or subcutaneous routes. Thus, the compounds disclosed herein may be administered systemically, e.g., orally, in combination with a pharmaceutically acceptable carrier, e.g., an inert diluent or an absorbable edible carrier. They may be enclosed in hard or soft shell gelatin capsules, may be compressed into tablets, or may be mixed directly with the food in the patient's diet. For oral therapeutic administration, the active compounds may be combined with one or more excipients and used in the form of ingestible tablets, buccal tablets, troches, capsules, elixirs, suspensions, syrups, wafers, and the like. Such compositions and formulations may contain at least 0.1% active compound. The percentage of the compositions and formulations may vary and may conveniently be between about 2% and about 60% by weight of a given unit dosage form. The amount of active compound in such therapeutically useful compositions is that amount which will achieve an effective dosage level (e.g., when measured systemically and/or locally after administration).

Tablets, troches, pills, capsules and the like containing one or more compounds of formula I further contain the following: binders, such as gum tragacanth, acacia, corn starch or gelatin; excipients such as dicalcium phosphate; disintegrants such as corn starch, potato starch, alginic acid and the like; lubricants such as magnesium stearate; and a sweetener such as sucrose, fructose, lactose or aspartame, or a flavoring agent such as peppermint, oil of wintergreen, or cherry flavoring may be added. When the unit dosage form is a capsule, it may contain, in addition to materials of the above type, a liquid carrier such as vegetable oil or polyethylene glycol. Various other materials may be present as coatings or may be present in physical form that otherwise improves the solid unit dosage form. For example, tablets, pills, or capsules may be coated with gelatin, wax, shellac, or sugar and the like. A syrup or elixir may contain the active compound, sucrose or fructose as a sweetening agent, methyl and propylparabens as preservatives, a dye and flavoring such as cherry or orange flavor. In general, any material used to prepare any unit dosage form should be pharmaceutically acceptable and substantially non-toxic in the amounts used. Furthermore, the active compounds may be incorporated into sustained release formulations and devices.

Compounds of the present disclosure (e.g., those of any of formulas I-IV) may also be administered intravenously or intraperitoneally by infusion or injection. Solutions of such compounds or pharmaceutically acceptable salts thereof can be prepared in water, optionally mixed with a non-toxic surfactant. Dispersions can also be prepared in glycerol, liquid polyethylene glycols, glyceryl triacetate and mixtures thereof, and in oils. Under ordinary conditions of storage and use, these formulations contain preservatives to prevent microbial growth. Pharmaceutical dosage forms suitable for injection or infusion may comprise sterile aqueous solutions or dispersions or sterile powders containing the active ingredient which are suitable for the extemporaneous preparation of sterile injectable or infusible solutions or dispersions, optionally encapsulated in liposomes. In all cases, the final dosage form should be sterile, fluid and stable under the conditions of manufacture and storage. The liquid carrier or vehicle may be a solvent or liquid dispersion medium including, for example, water, ethanol, polyols (e.g., glycerol, propylene glycol, liquid polyethylene glycol, and the like), vegetable oils or non-toxic glycerides and suitable mixtures thereof. Proper fluidity can be maintained, for example, by the formation of liposomes, by the maintenance of the required particle size in the case of dispersions, or by the use of surfactants. Prevention of microbial action can be achieved by various antibacterial and antifungal agents, such as parabens, chlorobutanol, phenol, sorbic acid, thimerosal (thimerosal), and the like. In many cases, it is preferable to include isotonic agents, for example, sugars, buffers, or sodium chloride. Prolonged absorption of the injectable compositions can be brought about by the use in the composition of agents delaying absorption, for example, aluminum monostearate and gelatin.

Sterile injectable solutions may be prepared by incorporating the active compound in the required amount in the appropriate solvent with various other ingredients enumerated above, as required, followed by filtered sterilization. In the case of sterile powders for the preparation of sterile injectable solutions, the preferred methods of preparation are vacuum-drying and freeze-drying techniques which yield a powder of the active ingredient plus any additional desired ingredient present in the previously sterile-filtered solution. For topical application, the compounds of the invention may be applied in pure form, for example when they are liquids. However, it will often be desirable to apply them to the skin in a composition or formulation in combination with a dermatologically acceptable carrier, which may be solid or liquid. Useful solid carriers include finely divided solids such as talc, clay, microcrystalline cellulose, silica, alumina and the like. Suitable liquid carriers include water, alcohols or glycols or water-alcohol/glycol blends in which the compounds of the present invention can be dissolved or dispersed at effective levels, optionally with the aid of non-toxic surfactants. Adjuvants such as fragrances and other antimicrobial agents may be added to optimize the properties of a given application. The resulting liquid composition may be applied from an absorbent pad for impregnating bandages and other dressings, or sprayed onto the affected area using a pump sprayer or an aerosol sprayer. Thickeners such as synthetic polymers, fatty acids, fatty acid salts and esters, fatty alcohols, modified celluloses or modified minerals may also be used with the liquid carrier to form coatable pastes, gels, ointments, soaps, and the like for direct application to the skin of a user. Examples of useful dermatological compositions that can be used to deliver the compounds of formula I to the skin are known in the art; see, for example, jacquet et al (U.S. Pat. No. 4,608,392), geria (U.S. Pat. No. 4,992,478), smith et al (U.S. Pat. No. 4,559,157), and Wortzman (U.S. Pat. No. 4,820,508). The effective dose of the compounds of formula I can be determined by comparing their in vitro and in vivo activity in animal models. Methods for extrapolating effective dosages in mice and other animals to humans are known in the art; see, for example, U.S. patent No. 4,938,949.

The amount of a compound disclosed herein, or an active pharmaceutically acceptable salt or derivative thereof, for use in therapy may vary not only with the particular salt selected, but also with the route of administration, the nature of the condition being treated, and the age and condition of the patient, and will ultimately be at the discretion of the attendant physician or clinician. The desired dosage may be conveniently presented in a single dosage form or in divided dosage forms for administration at appropriate intervals, for example in sub-dosage forms of two, three, four or more times daily. The sub-dose itself may be further divided into, for example, several discrete, loosely spaced administrations; such as multiple inhalations from an insufflator or by applying multiple drops to the eye.

As described herein, provided herein are pharmaceutical compositions comprising one or more compounds of formula I and/or conjugates thereof, and prepared for therapeutic and prophylactic use in mammals (e.g., humans). In some cases, such pharmaceutical compositions are formulated into oral dosage forms or injection dosage forms. Exemplary dosage forms are shown in table 1 below.

Table 1-dosage forms according to embodiments of the present disclosure

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Compositions comprising compounds of formulas I-IV, conjugates, and/or pharmaceutically acceptable salts thereof may be formulated in unit dosage forms, each dosage containing from about 5 to about 1,000mg (1 g), more typically from about 100mg to about 500mg, of the active ingredient. The term "unit dosage form" refers to physically discrete units suitable as unitary dosages for human subjects and other subjects, each unit containing a predetermined quantity of active material (i.e., a compound of formulas I-IV or a pharmaceutically acceptable salt thereof) calculated to produce the desired therapeutic effect in association with a suitable pharmaceutical excipient.

An effective amount of the active agent (i.e., a compound or conjugate of formulas I-IV, or a pharmaceutically acceptable salt of any of the foregoing) is typically provided at a dosage level of about 0.01mg/kg to about 1000mg/kg body weight per day, or any range therein. In some cases, the range is about 0.05 to about 500mg/kg body weight per day, or any range therein. In some cases, about 0.1 to about 250mg/kg body weight per day, or any range therein. In some cases, about 0.1 to about 100mg/kg body weight per day, or any range therein. In one example, the range may be about 0.1 to about 50.0mg/kg body weight per day, or any amount or range therein. In another example, the range may be about 0.01 to about 15.0mg/kg body weight per day, or any range therein. In yet another example, the range may be about 0.05 to about 7.5 mg/kg of body weight per day, or any amount within the range therein. In yet another example, the range may be about 0.1 to about 5.0mg/kg of body weight per day, or any amount within the range therein. Pharmaceutical compositions comprising compounds of formulas I-IV or pharmaceutically acceptable salts of any of the foregoing compounds may be administered on a regimen of 1 to 4 times per day or in a single daily dose.

VI methods of use

Provided herein are methods of treating a disease or pathological condition in a subject in need thereof using one or more compounds of formulas I-IV, conjugates thereof, and/or pharmaceutical compositions comprising such compounds. In some embodiments, such methods may comprise administering to a subject in need thereof a therapeutically effective amount of a compound, conjugate or pharmaceutical composition of formulas I-IV of the present disclosure, thereby treating such diseases or conditions in the subject. In some cases, the subject is a mammal, such as a human or rodent. Diseases and/or conditions that may be treated using the compounds of the present disclosure include, but are not limited to, cancer, viral or bacterial infections.

In some embodiments, the cancer, including but not limited to a tumor, metastasis, or other disease or disorder characterized by abnormal cells characterized by uncontrolled cell growth, is treated or inhibited by administering a compound of the present disclosure or a conjugate thereof as disclosed herein. In some embodiments, the subject has previously received a cancer treatment. In some embodiments, the previous treatment is surgery, radiation therapy, administration of one or more anticancer agents, or any combination of the foregoing.

In any of the methods described herein, the cancer is selected from the group consisting of: adenocarcinomas, adrenocortical carcinomas, adrenal neuroblastomas, anal squamous cell carcinomas, appendicular adenocarcinomas, bladder urothelial carcinomas, cholangiocarcinomas, bladder carcinomas, bladder urothelial carcinomas, bone chordoma, lymphocytic chronic myelogenous leukemia, non-lymphocytic acute myelogenous leukemia, myeloproliferative diseases, myelomas, osteosarcomas, brain astrocytomas, glioblastomas, brain medulloblastomas, meningiomas, brain oligodendrogliomas, adenoid cystic carcinomas, breast carcinomas, ductal carcinoma of the breast, invasive lobular carcinoma of the breast, breast carcinoma of the breast, cervical neuroendocrine carcinoma, cervical squamous cell carcinoma, colonic adenocarcinoma, colon carcinoid carcinoma, duodenal adenocarcinoma, endometrial tumors, esophageal adenocarcinoma, esophageal carcinoma, gastric carcinoma, invasive ductal carcinoma of the breast, invasive lobular carcinoma of the breast, invasive carcinoma of the breast, cervical carcinoma of the cervical carcinoma, endometrial tumor, esophageal carcinoma, gastric carcinoma, and gastric carcinoma intraocular melanoma, intraocular squamous cell carcinoma, tear duct carcinoma, serous fallopian tube carcinoma, gall bladder adenocarcinoma, gall bladder glomeruloma, gastroesophageal junction adenocarcinoma, head and neck adenoid cystic carcinoma, head and neck neuroblastoma, head and neck squamous cell carcinoma, renal colored cell carcinoma (kidney chromophore carcinoma), renal medullary carcinoma, renal cell carcinoma, renal papillary carcinoma, renal sarcoidosis, renal urothelial carcinoma, renal carcinoma, lymphoblastic leukemia chronic lymphocytic leukemia, hepatobiliary cell carcinoma, hepatocellular carcinoma, liver cancer, lung adenocarcinoma, lung adenosquamous carcinoma, atypical lung carcinoma, lung carcinoma sarcoma, large lung cell neuroendocrine carcinoma, lung non-small cell lung carcinoma, lung sarcoma, pulmonary sarcoidosis, small lung cell carcinoma, small lung cell undifferentiated carcinoma, squamous lung cell carcinoma, squamous cell carcinoma of upper respiratory tract, digestive tract carcinoma, diffuse large B-cell lymphoma of lymph nodes, follicular lymphoma of lymph nodes, mediastinal B-cell lymphoma of lymph nodes, lymphomatous plasmablasts of lung adenocarcinoma, lymphomatous follicular lymphoma, non-Hodgkins, nasopharyngeal and sinus undifferentiated carcinoma, ovarian carcinoma sarcoma, ovarian clear cell carcinoma, ovarian epithelial carcinoma, ovarian granuloma, ovarian serous carcinoma, pancreatic ductal adenocarcinoma, pancreatic neuroendocrine carcinoma, peritoneal mesothelioma, peritoneal serous carcinoma, placental choriocarcinoma, pleural mesothelioma, prostate adenocarcinoma, prostate carcinoma, rectal adenocarcinoma, rectal squamous cell carcinoma, skin attachment carcinoma, basal skin carcinoma, skin melanoma, skin merck cell carcinoma (skin Merkel cell carcinoma), skin squamous cell carcinoma, small intestine adenocarcinoma, small intestine intestinal tract mesothelioma (GIST), small intestine mesothelioma large intestine/colon cancer, large intestine adenocarcinoma, soft tissue angiosarcoma, soft tissue ewing sarcoma (soft tissue Ewing sarcoma), soft tissue vascular endothelial tumor, soft tissue inflammatory myofibroblastic tumor, soft tissue leiomyosarcoma, soft tissue liposarcoma, soft tissue neuroblastoma, soft tissue paraganglioma, perivascular epithelial cell tumor, soft tissue sarcoma, soft tissue synovial sarcoma, gastric adenocarcinoma, diffuse gastric adenocarcinoma, intestinal gastric adenocarcinoma, gastric leiomyosarcoma, thymus carcinoma, lymphocytic thymoma, papillary thyroid carcinoma, primary unknown adenocarcinoma, primary unknown carcinoma, primary unknown malignant tumor, lymphoid tumor, primary unknown melanoma, primary unknown sarcoma-like carcinoma, primary unknown squamous cell carcinoma, unknown undifferentiated neuroendocrine carcinoma, undifferentiated small cell carcinoma, uterine carcinoma sarcoma, endometrial adenocarcinoma, endometrioid endometrial adenocarcinoma, papillary serous endometrial adenocarcinoma, and uterine leiomyosarcoma, which are not originally identified.

Methods provided herein include methods of treating one or more diseases in a subject in need thereof. Some embodiments provide methods of treating cancer in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a compound of formulas I-IV, a pharmaceutically acceptable salt thereof, a conjugate thereof, or a pharmaceutical composition thereof. Some embodiments provide methods of inducing an anti-tumor immune response in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a compound of formulas I-IV, a pharmaceutically acceptable salt thereof, a conjugate thereof, or a pharmaceutical composition thereof.

Some embodiments herein provide methods of treating cancer in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a compound of formulae I-IV, or a pharmaceutically acceptable salt or conjugate thereof, in combination with another anti-cancer therapy (e.g., surgery and radiation therapy) and/or an anti-cancer agent (e.g., an immunotherapy such as nivolumab (nivolumab) or pembrolizumab). The compounds of formulas I-IV or conjugates thereof may be administered to a subject before, during, or after administration of the anti-cancer therapy and/or anti-cancer agent. In some embodiments, the compounds of formulas I-IV described herein may be administered to a subject after radiation therapy and/or post-operatively.

Some embodiments provide methods for delaying or preventing acquired resistance to an anti-cancer agent comprising administering to a subject (to a patient at risk of developing or having acquired resistance to an anti-cancer agent) a therapeutically effective amount of a compound of formulas I-IV, a pharmaceutically acceptable salt thereof, or a conjugate thereof. In some embodiments, a dose of an anti-cancer agent is administered to a patient (e.g., substantially simultaneously with administration of a dose of a compound of formulas I-IV, a pharmaceutically acceptable salt thereof, or a conjugate thereof to a patient).

Some embodiments provide methods of delaying and/or preventing the progression of cancer that is resistant to an anti-cancer agent in a subject comprising administering to the subject a therapeutically effective amount of a compound of formulas I-IV, a pharmaceutically acceptable salt thereof, or a conjugate thereof, before, during, or after administration of the therapeutically effective amount of the anti-cancer agent.

The compounds of formulas I-IV and/or conjugates thereof are useful for inhibiting proliferation of cancer cells, causing apoptosis of cancer cells, for increasing phagocytosis of cancer cells, and/or for treating cancer in a subject in need thereof. In some embodiments, the cancer is as described herein. In some embodiments, the subject has previously received a cancer treatment. In some embodiments, the previous treatment is surgery, radiation therapy, administration of one or more anticancer agents, or any combination of the foregoing. In some embodiments, the subject has discontinued prior treatment, for example, due to unacceptable or intolerable side effects, wherein the prior treatment is too toxic, or wherein the subject develops resistance to the prior treatment. Some embodiments provide methods for delaying or preventing a disease or disorder comprising administering to a subject (to a patient at risk of having the disease or disorder) a therapeutically effective amount of a compound of formulas I-IV or a pharmaceutically acceptable salt thereof, and a vaccine against the disease or disorder. In some embodiments, the disease or disorder is cancer, as described herein. In some embodiments, the disease or disorder is a viral pathogen. In some embodiments, the vaccine is administered subcutaneously. In some embodiments, the vaccine is administered intramuscularly. In some embodiments, the compounds of formulas I-IV, pharmaceutically acceptable salts thereof, or conjugates thereof, and the vaccine are administered by the same route (e.g., the compounds of formulas I-IV, or pharmaceutically acceptable salts thereof, and the vaccine are all administered subcutaneously). In some embodiments, the compounds of formulas I-IV, pharmaceutically acceptable salts thereof, or conjugates thereof, and the vaccine are administered by different routes. In some embodiments, the vaccine and the compound of formulas I-IV, or a pharmaceutically acceptable salt thereof, are provided in a single formulation. In some embodiments, the vaccine and the compound of formulas I-IV, or a pharmaceutically acceptable salt thereof, are provided in separate formulations. In some embodiments, the compounds of formulas I-IV described herein are present in salt form when used in therapy. In some embodiments, the salt is a pharmaceutically acceptable salt.

VII Compounds Activity and test

Provided herein are various experimental procedures for the synthesis, evaluation, and use of compounds (e.g., those of formulas I-IV) and conjugates of the present disclosure.

Screening compounds for their binding ability to TLR-7/8 expressing cells: toll-like receptor (TLR) -7 or TLR-8 positive cell lines and HEK-Blue TLR cells can be used in the screening assay. The HEK-Blue TLR cells may be engineered HEK293 cells that stably express the TLR gene and an inducible NF-kB-SEAP (secreted embryonic alkaline phosphatase) reporter gene. Binding of a ligand (e.g., a compound of formulas I-IV) to a TLR in HEK-Blue cells can induce SEAP with pNPP as a phosphatase substrate to turn Blue. In some cases, a compound of the present disclosure may be added in triplicate to a temperature of 37 ℃ and 5% co, for example, at a concentration of about 20nmol/ml or about 5.2nmol/ml ₂ HEK-Blue-TLR7 or TLR8 cells cultured under conditions. After a period of time, for example 24 hours, a volume (e.g., 5 μl) of supernatant from each experiment may be mixed with about 200 μl of detection medium containing pNPP. After one hour, SEAP activity can be read as OD at 650nm using a microplate reader to determine the binding affinity of the test compound.

TLR7/8 modulator NF- κb reporter assay: human Embryonic Kidney (HEK) cells stably transfected with human TLR-7 or TLR-8 and NF- κb-reactive Secreted Embryonic Alkaline Phosphatase (SEAP) genes (HEK-TLR-7 and-8) can be purchased and then stimulated with 30 μm compound (e.g., compound of formula I-IV) in DMEM containing 10% fbs and 0.01% normocin (invitogen) in 96-well plates for about 24h. Subsequently, a volume (e.g., about 20 μl) of supernatant from each well can be incubated with the Quanti-blue substrate solution (invitogen) at 37 ℃ for about 1h, and absorbance can be read at 650nm wavelength.

Measurement of pro-inflammatory cytokines by flow bead assay (Cytometric Bead Assay): bone marrow derived dendritic cells (BMDCs) can be generated by isolating a single cell suspension of bone marrow from the femur of a C57BL/6 mouse (6-8 weeks old). Then 0.83% NH can be used ₄ Cl、0.1％KHCO ₃ And 0.009% lysed erythrocytes. 500 ten thousand cells can be seeded in full RPMI medium (Invitrogen, grand Island, N.Y.) in each well of a 6-well plate supplemented with 20ng/ml granulocyte-macrophage colony stimulating factor (PeproTech, rocky Hill, N.J.) in mice. After a few days of incubation, BMDC may be stimulated with 30. Mu.M of a compound (e.g., a compound of formulas I-IV) for 3 days. Subsequently, a volume of about 25. Mu.L of supernatant can be removed and TNF. Alpha., IL-12p40, IL-1. Beta. And IL-10 can be determined using a flow cytometric bead array according to the manufacturer's instructions (e.g., BD Bioscience, san Jose, calif.). Controls can be performed by adding the medium and carrier without the drug.

Evaluation of effects on human monocyte-derived dendritic cells (MoDC): moDCs can be seeded into 96-well plates and stimulated with several (e.g., 5 or more) different concentrations of one or more compounds of formula I at various concentrations (e.g., about 0.3, 0.5, 1.3, 5.2, and 20 nmol/cell culture medium) for about 48 hours and performed in triplicate. For immunostaining and flow cytometry analysis, and about 48 hours after stimulation, cells can be stained with anti-HLA-DR, CD11c, CD-86, CD80, CD83, CD8a, CD123, combinations thereof, and related isotype controls, followed by analysis, including cytokine analysis.

In vivo evaluation of IL-6 levels: 8-12 week old TLR7 mutant mice and C57BL/6j mice can be used to introduce the TLR7 mutant gene into 129S1/Sv derived from CJ7 embryonic stem cells. This cell line can be backcrossed ten times with C57BL/6Ncr, and TLR7RNA expression was not detected in bone marrow-derived macrophages. Single cell suspensions of spleen cells from C57BL/6j or TLR7 mutant mice can then be isolated. Splenocytes in culture media may be treated in triplicate with varying concentrations of one or more compounds of the present disclosure. For example, supernatants from the culture media can be harvested 12 hours and 24 hours after treatment. Flow microbead array (cytometric bead array) (BD Biosciences, san Jose, calif.) can be used to measure IL-6 levels.

proliferation of pmel CD8 and ifnγ production: single cell suspensions of B MDC cells from C56BL/6j mice and spleen cells from pmel mice can be treated in triplicate with about 3.5 μg human gp100 peptide per well (gp 100 peptide sequence: CALLAVGATKVPR-NQDWLGVSRQL RTK, genScript, piscataway, NJ) and test compound (e.g., compound of formula I) at a concentration of about 10.4 nmol/ml. BMDC cells can then be washed twice with complete RPMI medium and then co-cultured with CFSE labeled pmel CD8 splenocytes and isolated from pmel splenocytes at a DC/CD8 ratio of 1:3 using a cd8+ T cell isolation kit. Four days after co-cultivation, the supernatant can be harvested and subsequently subjected to ifnγ measurement. The cell pellet may be washed and stained with a fluorescently labeled antibody.

IL-2 production by OT-I cells after stimulation with compounds of formulas I-IV: single cell suspensions of C57BL/6j can be treated in triplicate with a concentration of about 20.8nmol/ml of test compound (e.g., one of formula I) with and without 15 μg ovalbumin added per well. Four days later, the cells may be washed twice with complete RPMI medium and cultured with isolated OT-I CD 8T cells. Four days after co-cultivation, the supernatant can be harvested and tested. CBA may be performed for IL-2 production.

Description of the embodiments

For further explanation, additional non-limiting embodiments of the present disclosure are set forth below.

Embodiment P1: a compound of formula I:

(I)

wherein:

fused ring a is selected from the group consisting of:

R ² is H, (C) ₁ -C ₆ ) Alkyl, (C) ₂ -C ₆ ) Alkenyl group (C) ₂ -C ₆ ) Alkynyl, (C) ₁ -C ₆ ) Alkanoyl, (C) ₁ -C ₆ ) Alkoxycarbonyl, (C) ₁ -C ₆ ) Alkanoyloxy, (C) ₃ -C ₆ ) Cycloalkyl, aryl, heteroaryl, heterocycle, NR ^g R ^h Or R is ^m R ⁿ NC(＝O)-，Any of (C) ₁ -C ₆ ) Alkyl, (C) ₂ -C ₆ ) Alkenyl group (C) ₂ -C ₆ ) Alkynyl, (C) ₁ -C ₆ ) Alkanoyl, (C) ₁ -C ₆ ) Alkoxycarbonyl, (C) ₁ -C ₆ ) Alkanoyloxy, (C) ₃ -C ₆ ) Cycloalkyl, aryl, heteroaryl, and heterocycle are optionally substituted with one or more groups independently selected from the group consisting of: hydroxy, halo, -SH, cyano, oxo, oxirane, (C) ₃ -C ₈ ) Cycloalkyl, aryl, heteroaryl, (C) ₁ -C ₆ ) Alkoxy, (C) ₁ -C ₆ ) Alkylthio and NR ^g R ^h ；

R ^b Is H or X-Y;

R ^e is H, (C) ₁ -C ₆ ) Alkyl or R ^aa ；

R ^f Is H or (C) ₁ -C ₆ ) An alkyl group;

x is a linking group; and is also provided with

Y is a peptide, protein or maleimide;

wherein rings B and C in formula I can optionally be one or more independently selected from halo, hydroxy, nitro, (C) at one or more carbons ₁ -C ₆ ) Alkyl, (C) ₁ -C ₆ ) Alkenyl group (C) ₁ -C ₆ ) Alkynyl, (C) ₁ -C ₆ ) Alkoxy radicalRadical (C) ₁ -C ₆ ) Alkanoyl, (C) ₁ -C ₆ ) Alkanoyloxy, (C) ₁ -C ₆ ) Alkoxycarbonyl, trifluoromethyl, trifluoromethoxy, cyano and NR ^p R ^q Further substitution of the groups of the group;

R ^aa Is aryl optionally substituted with one or more groups independently selected from the group consisting of: hydroxy, halogen, nitro, cyano, ("A")C ₁ -C ₆ ) Alkoxy and optionally substituted with one or more halo (C ₁ -C ₆ ) An alkyl group;

or a pharmaceutically acceptable salt thereof.

Embodiment P2: the compound of embodiment P1, wherein R ⁴ Is R ^k -C(＝O)-、R ^k -O-C(＝O)-、R ^k -S(O) ₂ -O-、R ^c R ^d NC(＝O)-、R ^c R ^d NS(O) ₂ -、R ^e C(＝O)N(R ^e ) -or R ^e S(O) ₂ NR ^f 。

Embodiment P3: the compound of embodiment P1 or P2, wherein R ⁴ Is R ^e S(O) ₂ NR ^f -and R ^e Is R ^w 。

Embodiment P4: the compound of embodiment P1 or P2, wherein R ⁴ Is R ^k -C(＝O)-、R ^k -O-C (=o) -or R ^k -S(O) ₂ -O-。

Embodiment P5: the compound of embodiment P1, wherein R ⁴ Is at the 2-position by R ^k -C(＝O)-、R ^k -O-C(＝O)-、R ^k -S(O) ₂ -O-、R ^c R ^d NC(＝O)-、R ^c R ^d NS(O) ₂ -、R ^e C(＝O)N(R ^e ) -or R ^e S(O) ₂ NR ^f -substituted 1-ethylene.

Embodiment P6: the compound of any one of embodiments P1, P2, P4 or P5, wherein R ^k Is H, (C) ₁ -C ₆ ) Alkyl, (C) ₂ -C ₆ ) Alkenyl group (C) ₂ -C ₆ ) Alkynyl, (C) ₃ -C ₈ ) Cycloalkyl, trifluoromethyl, aryl or aryl (C) ₁ -C ₆ ) Alkyl groups, each of which (C ₁ -C ₆ ) The alkyl group being optionally substituted by one or more halo groups, (C) ₁ -C ₆ ) Alkanoyloxy, (C) ₁ -C ₆ ) Alkoxy or (C) ₃ -C ₈ ) Cycloalkyl substitution.

Embodiment P7: the compound of any one of embodiments P1 to P6, wherein ring a is selected from the group consisting of:

embodiment P8: the compound of embodiment P7 wherein R ¹ And R is ³ Each independently is H or optionally substituted (C) ₁ -C ₆ ) An alkyl group.

Embodiment P9: the compound of embodiment 7P or P8 wherein R ² Is H, (C) ₁ -C ₆ ) Alkyl, (C) ₂ -C ₆ ) Alkenyl group (C) ₂ -C ₆ ) Alkynyl, (C) ₁ -C ₆ ) Alkanoyl, (C) ₁ -C ₆ ) Alkoxycarbonyl, (C) ₁ -C ₆ ) Alkanoyloxy, (C) ₃ -C ₆ ) Cycloalkyl, aryl, heteroaryl, heterocycle, NR ^g R ^h Or R is ^m R ⁿ NC (=o) -, any of which (C) ₁ -C ₆ ) Alkyl, (C) ₂ -C ₆ ) Alkenyl group (C) ₂ -C ₆ ) Alkynyl, (C) ₁ -C ₆ ) Alkanoyl, (C) ₁ -C ₆ ) Alkoxycarbonyl, (C) ₁ -C ₆ ) Alkanoyloxy, (C) ₃ -C ₆ ) Cycloalkyl, aryl, heteroaryl, and heterocycle are optionally substituted with one or more groups independently selected from the group consisting of: hydroxy, halo, -SH, cyano, oxo, oxirane, (C) ₃ -C ₈ ) Cycloalkyl, aryl, heteroaryl, (C) ₁ -C ₆ ) Alkoxy, (C) ₁ -C ₆ ) Alkylthio and NR ^g R ^h 。

Embodiment P10: the compound of any one of embodiments P1 to P9, wherein ring a is selected from the group consisting of:

embodiment P11: the method according to any one of embodiments P1 to P10Wherein R is a compound of formula (I) ² Is H or optionally substituted (C) ₁ -C ₆ ) An alkyl group.

Embodiment P12: the compound of any one of embodiments P9 to P11, wherein ring a is

Embodiment P13: the compound of any one of embodiments P9 to P11, wherein ring a is

Embodiment P14: the compound of any one of embodiments P9 to P11, wherein ring a is

Embodiment P15: the compound of any one of embodiments P9 to P11, wherein ring a is

Embodiment P16: a compound of formula II:

(II)

Or a pharmaceutically acceptable salt thereof,

wherein:

R ⁴ Is R ^k -C(＝O)-、R ^k -O-C (=o) -or R ^k -S(O) ₂ -O-；

R ^b Is H or X-Y;

x is a linking group; and is also provided with

Y is a peptide, protein or maleimide.

Embodiment P17: the compound of embodiment P16, wherein R ² Is H or optionally substituted (C) ₁ -C ₆ ) An alkyl group.

Embodiment P18: the compound of embodiment P16 or P17 wherein R ⁴ Is R ^k -C (=o) -or R ^k -O-C(＝O)-。

Embodiment P19: the compound of embodiment P18, wherein R ^k Is H or optionally substituted (C) ₁ -C ₆ ) An alkyl group.

Embodiment P20: the compound of any one of embodiments P16 to P19, wherein R ^a Is H or optionally substituted (C) ₁ -C ₆ ) An alkyl group.

Embodiment P21: the compound of any one of embodiments P16 to P20, wherein R ^b Is H or X-Y.

Embodiment P22: the compound of any one of embodiments P16 to P21 having the formula:

embodiment P23: the compound of any one of embodiments P1 to P7, wherein ring a is:

wherein the method comprises the steps of

R ³ Is H, halo, hydroxy, (C) ₁ -C ₆ ) Alkyl, (C) ₂ -C ₆ ) Alkenyl group (C) ₂ -C ₆ ) Alkynyl, (C) ₁ -C ₆ ) Alkanoyl, (C) ₁ -C ₆ ) Alkoxycarbonyl, (C) ₁ -C ₆ ) Alkanoyloxy, (C) ₃ -C ₆ ) Cycloalkyl, aryl, heteroaryl, heterocycle, NR ^g R ^h Or R is ^m R ⁿ NC (=o) -, any of which (C) ₁ -C ₆ ) Alkyl, (C) ₂ -C ₆ ) Alkenyl group (C) ₂ -C ₆ ) Alkynyl, (C) ₁ -C ₆ ) Alkanoyl, (C) ₁ -C ₆ ) Alkoxycarbonyl, (C) ₁ -C ₆ ) Alkanoyloxy, (C) ₃ -C ₆ ) Cycloalkyl, aryl, heteroaryl, and heterocycle are optionally substituted with one or more groups independently selected from the group consisting of: hydroxy, halo, -SH, cyano, oxo, oxirane, (C) ₃ -C ₈ ) Cycloalkyl, aryl, heteroaryl, (C) ₁ -C ₆ ) Alkoxy, (C) ₁ -C ₆ ) Alkylthio and NR ^g R ^h 。

Embodiment P24: a compound of formula III or formula IV:

Or a pharmaceutically acceptable salt thereof,

wherein:

R ² is H, (C) ₁ -C ₆ ) Alkyl, (C) ₂ -C ₆ ) Alkenyl group (C) ₂ -C ₆ ) Alkynyl, (C) ₁ -C ₆ ) Alkanoyl, (C) ₁ -C ₆ ) Alkoxycarbonyl, (C) ₁ -C ₆ ) Alkanoyloxy, (C) ₃ -C ₆ ) Cycloalkyl, aryl, heteroaryl, heterocycle,NR ^g R ^h Or R is ^m R ⁿ NC (=o) -, any of which (C) ₁ -C ₆ ) Alkyl, (C) ₂ -C ₆ ) Alkenyl group (C) ₂ -C ₆ ) Alkynyl, (C) ₁ -C ₆ ) Alkanoyl, (C) ₁ -C ₆ ) Alkoxycarbonyl, (C) ₁ -C ₆ ) Alkanoyloxy, (C) ₃ -C ₆ ) Cycloalkyl, aryl, heteroaryl, and heterocycle are optionally substituted with one or more groups independently selected from the group consisting of: hydroxy, halo, -SH, cyano, oxo, oxirane, (C) ₃ -C ₈ ) Cycloalkyl, aryl, heteroaryl, (C) ₁ -C ₆ ) Alkoxy, (C) ₁ -C ₆ ) Alkylthio and NR ^g R ^h ；

R ⁴ Is R ^k -C(＝O)-、R ^k -O-C (=o) -or R ^k -S(O) ₂ -O-；

R ^b Is H or X-Y;

x is a linking group; and is also provided with

Y is a peptide, protein or maleimide.

Embodiment P25: the compound of embodiment P24, wherein R ¹ And R is ² Independently is H or can optionally be covered (C) ₁ -C ₃ ) Alkyl, substituted or unsubstituted (C ₃ -C ₈ ) Cycloalkyl or substituted or unsubstituted aryl substituted (C ₁ -C ₆ ) An alkyl group.

Embodiment P26: the compound of embodiment P24 or P25, wherein R ⁴ Is R ^k -O-C (=o) -or R ^k -S(O) ₂ -O-。

Embodiment P27: the compound of embodiment P24 or P25 selected from the group consisting of:

embodiment P28: the compound of any one of embodiments P1 to P21 or P23 to P26, wherein X is (C ₁ -C ₆ ) Alkyl, (C) ₂ -C ₆ ) Alkenyl or (C) ₁ -C ₆ ) Alkynyl, said (C ₁ -C ₆ ) Alkyl, (C) ₂ -C ₆ ) Alkenyl or (C) ₁ -C ₆ ) Alkynyl groups are optionally substituted with oxo groups.

Embodiment P29: the compound of any one of embodiments P1 to P21 or P23 to P26, wherein X is selected from the group consisting of:

and wherein n is 2, 3, 4, 5 or 6.

Embodiment P30: the compound of any one of embodiments P1 to P21, P23 to P26, or P28 to P29, wherein Y is maleimide.

Embodiment P31: the compound of any one of embodiments P1 to P21, P23 to P26, or P28 to P29, wherein Y is an antigen.

Embodiment P32: the compound of embodiment P31, wherein the antigen is associated with a bacterium or virus, and wherein the virus is selected from the group consisting of influenza virus, HIV, and HCV.

Embodiment P33: the compound of any one of embodiments P1 to P21, P23 to P26, or P28 to P29, wherein Y is an antigen associated with a tumor cell or tumor cell lysate.

Embodiment P34: the compound of any one of embodiments P31 to P33, wherein Y is an antigen comprising a peptide sequence comprising cysteine, lysine, or both.

Embodiment P35: the compound of any one of embodiments P1 to P34, wherein the compound is any one of compounds 1 or 22 to 26.

Embodiment P36: a pharmaceutical composition comprising a compound according to any one of embodiments P1 to P35 and a pharmaceutically acceptable diluent or carrier.

Embodiment P37: a method of treating a pathological condition in an animal comprising administering to the animal a compound according to any one of embodiments P1 to P35, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition according to embodiment P36.

Embodiment P38: a method of stimulating an immune response in an animal comprising administering to the animal a compound according to any one of embodiments P1 to P35, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition according to embodiment P36.

Embodiment P39: a method of treating cancer in an animal comprising administering to the animal a compound according to any one of embodiments P1 to P35, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition according to embodiment P36.

Embodiment P40: a method of synthesizing a compound of any one of embodiments P1 to P35, the method comprising performing a reaction according to any one of reaction schemes 1-5.

Examples

The following examples are given to illustrate various embodiments of the disclosure and are not meant to limit the disclosure in any way. The present examples and methods described herein presently represent some embodiments, are exemplary, and are not intended to limit the scope of the present disclosure. Variations and other uses thereof will occur to those skilled in the art that are encompassed within the spirit of the disclosure as defined by the scope of the claims.

Example 1

General synthetic procedure

This example shows the synthesis of compounds of formulas I and II.

General experimental conditions: all solvents were used as received from commercial suppliers except DMF, which was purified by 2 alumina columns in a solvent purification system. 2-amino-4-bromobenzoic acid was purchased from AstaTech. Pd2 (dba) ₃ And triethylamine were purchased from Chem-Impex. XPhos is available from TCI America. Phosphorus pentasulfide was purchased from Acros Organics. All other general chemicals were purchased from Sigma-Aldrich, macron, alfa Aesar, fisher Chemical, or synthesized using the cited literature schemes. Thin Layer Chromatography (TLC) on 0.25mm hard silica G plates; by usingThe developed plate was observed with a hand-held UV lamp. The silica used for chromatography is from SilicicleP60. Recording in said solvent on a Varian600MHz or Varian 400MHz spectrometer ¹ H and ¹³ c NMR spectrum; peaks are recorded as chemical shifts (multiplicity, J-coupling, in Hz, proton number). High resolution mass spectra were obtained on a Agilent TOF II TOF/MS instrument equipped with ESI or APCI interfaces.

Example 2

Synthesis of 4-bromo-2- ((2-nitroethylene) amino) benzoic acid (3)

2-amino-4-bromobenzoic acid (2, 3.0g,13.89mmol,1 eq.) was dissolved in a 1:1 mixture of water: dioxane (90 mL) at 50 ℃. The addition of nitroaldoxime (methacrylic acid) (40 mL, about 28mmol, about 2 eq.) resulted in immediate precipitation of an off-white solid. The solution was cooled to 25 ℃ and stirred for 18 hours. The pale yellow precipitate was filtered and rinsed with water until the filtrate was clear. The filtered solid was dried in a vacuum oven to constant mass. The crude yellow solid 3 was used without characterization.

Example 3

Nitroglyoxime synthesis

Sodium hydroxide (6.72 g,168 mmol) was dissolved in water (12 mL) and maintained at 25 ℃. Nitromethane (3 mL,56 mmol) was added dropwise to the aqueous sodium hydroxide solution with rapid stirring and heated to 35℃for 10 minutes. The reaction was cooled to 25 ℃ and another portion of nitromethane (3 ml,56 mmol) was added while carefully maintaining the temperature at 35 ℃ or below 35 ℃. After rapid stirring for 10 minutes, the reaction was heated to 50 ℃ for 5 minutes and then allowed to cool to 25 ℃ for 10 minutes. The reaction mixture was poured into cold water (50 mL) and then concentrated hydrochloric acid was added. Additional water was added until the total volume was 80mL. The crude nitroglyoxime solution was used immediately in the next step, or frozen (-20 ℃) and then thawed carefully before being used in the reaction shown in example 2.

Example 4

Synthesis of 7-bromo-3-nitroquinolin-4-ol (4)

Crude 4-bromo-2- ((2-nitroethyleneamino) benzoic acid (3, 0.45g,1.57mmol,1 eq.) was added to a flask with stirring bar and acetic anhydride (5 mL). The suspension was heated to 100 ℃ for 45 minutes, or until completely dissolved. Anhydrous sodium acetate was added to the reaction at once while vigorously stirring. The reaction was heated to reflux for 2 hours and then allowed to cool to 25 ℃. The contents of the flask were diluted with cooled 80% aqueous acetic acid and the solids filtered. The solid was rinsed with acetic acid until the filtrate was clear. The solid was dried in a vacuum oven at 45 ℃ overnight to give a pale brown powder that was easily peeled from the filter paper. The crude solid 4 has limited solubility in most solvents and therefore can proceed without characterization.

Example 5

Synthesis of 3-amino-7-bromo-4-hydroxyquinoline (5)

To a solution of crude 7-bromo-3-nitroquinolin-4-ol (0.34 g,1.26 mmol) in anhydrous DMF (16 mL) was added sodium dithionite (1.0 g). The reaction mixture was taken up in N ₂ Stirring was carried out at 25℃for 18 hours. DMF was removed in vacuo and the crude solid was suspended in ethyl acetate (15 mL) and then washed with hot water (35 mL). The mixture was stirred and heated, along with saturated aqueous sodium carbonate (10 mL), until all remaining sodium dithionite had decomposed. The organics were separated from the aqueous layer and extracted with ethyl acetate (4 x 10 ml). The organics were combined and concentrated in vacuo. The crude compound was passed through a silica column 85:9:3:3etoac: meoh: h ₂ O:Et ₃ N purification gave 24.8% yield as an off-white solid: ¹ H NMR((CD ₃ ) ₂ SO,400MHz)δ11.48(br s,1H),8.01(d,J＝8.8Hz,1H),7.68(s,1H),7.55(s,1H),7.29(d,J＝8.8Hz,1H),4.72(br s,2H)。

example 6

Synthesis of 7-bromo-2-butylthiazolo [4,5-c ] quinoline (6)

Direction N ₂ To a solution of purged 3-amino-7-bromo-4-hydroxyquinoline (0.07 g,0.29mmol,1 eq.) in anhydrous pyridine (3.4 mL) was added pentanoyl chloride (0.044 g,0.37mmol,1.25 eq.) and heated to 60℃in a pressure vessel for 2 hours. The reaction was cooled to 25 ℃ and placed again in N ₂ And flow down. Phosphorus pentasulfide (0.14 g,0.32mmol,1.1 eq.) was added and the reaction was heated to 105 ℃ in a pressure vessel for 18 hours. The reaction was cooled to 25 ℃ and aqueous sodium carbonate (10 mL) and ethyl acetate (10 mL) were added. The mixture was separated and the aqueous layer was extracted with ethyl acetate (2 x 5 ml) (note that traces of hydrogen sulfide in organics were present, which was typically removed with gentle vacuum with gentle heating for 2 hours). The organics were combined and concentrated in vacuo and purified via silica column 1:99meoh:dcm to give a yellow waxy solid in 86.1% yield: ¹ H NMR(CDCl ₃ 400 MHz) delta 9.43 (s, 1H), 8.43 (d, j=1.6 hz, 1H), 7.84 (d, j=8.4 hz, 1H), 7.73 (dd, j=8.4, 1.6hz, 1H), 3.23 (t, j=7.6 hz, 2H), 1.94 (quintuples, j=7.6 hz, 2H), 1.51 (app-six-fold peaks, j=7.6, 7.2hz, 2H), 1.01 (t, j=7.2 hz, 3H).

Example 7

Synthesis of 7-bromo-2-butylthiazolo [4,5-c ] quinoline 5-oxide (7)

To 7-bromo-2-butylthiazolo [4,5-c]Quinoline (0.075 g,0.23mmol,1 eq.) in dichloromethane (3mL) was added 53% m-chloroperoxybenzoic acid (0.27 g,0.82mmol,3.5 eq.). It was stirred at 25℃for 15 hours. TLC indicated almost complete reaction. The reaction mixture was washed with saturated aqueous sodium carbonate (10 mL), diluted with another portion of dichloromethane (10 mL) and water (10 mL), the mixture was separated, the aqueous layer was extracted with DCM (2×10 mL), and the organics were combined and concentrated in vacuo. The crude solid was purified by silica column 1:99-2:98meoh:dcm (linear gradient) to give a white powder in 92.7% yield: ¹ H NMR(CDCl ₃ 400 MHz) δ9.19 (s, 1H), 9.11 (s, 1H), 7.84 (s, 2H), 3.20 (t, j=7.6 hz, 2H), 1.92 (quintuple peak, j=7.6 hz, 2H), 1.51 (six-tuple peak, j= 7.6,2H), 1.00 (t, j=7.6 hz, 3H).

Example 8

Synthesis of N- (7-bromo-2-butylthiazolo [4,5-c ] quinolin-4-yl) benzamide (8)

To 7-bromo-2-butylthiazolo [4,5-c]To a solution of quinoline 5-oxide (0.073 g,0.22mmol,1 eq.) in methylene chloride (8 mL) was added benzoyl isocyanate (0.127 g,0.86mmol,4 eq.). After complete dissolution of the benzoyl isocyanate, the solution became pale yellow and was stirred at 25 ℃ for 18 hours. The reaction mixture was concentrated in vacuo and purified via silica column 0:100-1:99meoh:dcm (linear gradient) to give a white solid in 73.4% yield: ¹ H NMR(CDCl ₃ 400 MHz) delta 9.69 (br s, 1H), 8.41 (s, 1H), 8.08 (s, 1H), 8.06 (s, 1H), 7.70 (d, j=8.4 hz, 1H), 7.65-7.53 (m, 4H), 3.21 (t, j=7.6 hz, 2H), 1.92 (quintuple peak, j=7.6 hz, 2H), 1.53 (app-six-tuple peak, j=7.6, 7.2hz, 2H), 1.02 (t, j=7.2 hz, 3H). ¹³ C{ ¹ H}NMR(CDCl ₃ ,600MHz)δ173.0,164.6,145.4,144.3,140.7,139.3,134.7,132.4,132.2,129.2,128.9,127.7,125.5,122.9,120.2,34.0,31.9,22.3,13.8。(ESI+)：C ₂₁ H ₁₈ BrN ₃ OS[M+H]+calculated 440.0427, found 440.0437 (error 2.3 ppm).

Example 9

Synthesis of N- (2-butyl-7-phenoxycarbonylthiazolo [4,5-c ] quinolin-4-yl) benzamide (9)

A solution of formic acid (2 mL) in acetic anhydride (5 mL) was stirred at 35℃for 1 hour to yield acetic anhydride. N- (7-bromo-2-butylthiazolo [4, 5-c)]Quinolin-4-yl) benzamide (0.07 g,0.16mmol,1 eq.), phenol (0.061 g,0.65mmol,4.1 eq.), tris (dibenzylideneacetone) dipalladium (0) [ Pd) ₂ (dba) ₃ ](0.015 g,0.016mmol,10 mol%) and 9, 9-dimethyl-9H-xanthene-4, 5-diylbis (diphenylphosphine) [ XPhos ] ](0.021 g,0.037mmol,23 mol%) was added to the flask and the mixture was taken in N ₂ Suspended in toluene (4 mL) under reflux. The mixture was stirred for 1 minute and then pre-prepared acetic anhydride (75 μl,0.95mmol,6 eq.) was added in one portion while stirring followed immediately by triethylamine (100 μl,0.72mmol,4.5 eq.). The flask was capped with a septum and stirred for 2 minutes. Then another portion of triethylamine (100 μl) (1.44 mmol, total 9 equivalents) was added followed by another portion of acetic anhydride (30 μl) (1.33 mmol, total 8.3 equivalents). The reaction was stirred and heated to 85 ℃ for 18 hours, then cooled to 25 ℃ and diluted with ethyl acetate (5 mL). The crude reaction mixture was filtered through a small plug of silica, concentrated in vacuo, and purified through silica column 1:99meoh: dcm-4:96meoh: dcm to afford an off-white solid in 15.7% yield: ¹ H NMR(CDCl ₃ 600 MHz) δ9.76 (br s, 1H), 9.17 (s, 1H), 8.25 (dd, j=8.4, 1.8hz, 1H), 8.09 (d, j=7.8 hz, 2H), 7.94 (d, j=8.4 hz, 1H), 7.65-7.43 (m, 5H), 7.35-7.75 (m, 3H), 3.25 (t, j=7.8 hz, 2H), 1.95 (quintuple, j=7.8 hz, 2H), 1.54 (app-six-tuple, j=7.8, 7.2hz, 2H), 1.03 (t, j=7.2 hz, 3H). ¹³ C{ ¹ H}NMR(CDCl ₃ ,600MHz)δ174.4,164.7,164.4,151.0,145.4,143.1,140.3,140.2,134.6,132.4,129.9,129.6,129.51,129.50,128.9,127.6,126.1,125.9,124.8,121.6,34.0,31.9,22.3,13.7。(ESI+)：C ₂₈ H ₂₃ N ₃ O ₃ S[M+H]+calculated 482.1533, found 482.1572 (error 8.1 ppm).

Example 10

Synthesis of 4-amino-2-butyl-7-methoxycarbonylthiazolo [4,5-c ] quinoline (1)

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N- (2-butyl-7-phenoxycarbonyl thiazolo [4, 5-c)]Quinolin-4-yl) benzamide (0.010g, 0.020mmol) was added to a vial and suspended in anhydrous methanol (3 mL). The mixture was sonicated for 1 minute to break up any larger solid pieces. At N ₂ Under stream, concentrated sulfuric acid (3 drops, about 125 mg) was added, which rapidly dissolved the finely suspended solids. The vials were capped and stirred and heated at 60 ℃ for 15 hours. The reaction was cooled to 25 ℃ and neutralized with solid sodium carbonate powder, diluted with ethyl acetate (5 mL) and water (5 mL), and stirred for 10 minutes. The mixture was concentrated in vacuo and redissolved in equal amounts of ethyl acetate and water. The mixture was separated and the aqueous solution was extracted with ethyl acetate (3×5 mL). The organics were combined and concentrated in vacuo. The crude product was purified through silica column 25:75etoac:hexs to give the final product as a white solid in 68.7% yield: ¹ H NMR(CDCl ₃ 600 MHz) δ8.53 (d, j=1.2 hz, 1H), 8.03 (dd, j=8.4, 1.2hz, 1H), 7.81 (d, j=8.4 hz, 1H), 6.86 (br s, 2H), 3.98 (s, 3H), 3.20 (t, j=7.8 hz, 2H), 1.91 (app-quintessence, j=7.8, 7.2hz, 2H), 1.51 (hexa-quintessence, j=7.2 hz, 2H), 1.01 (t, j=7.2 hz, 3H). ¹³ C{ ¹ H}NMR(CDCl ₃ ,600MHz)δ174.5,166.2,151.0,141.5,138.7,138.1,131.5,125.5,125.2,124.6,121.5,52.6,33.9,31.8,22.2,13.7。(APCI+)：C ₁₆ H ₁₇ N ₃ O ₂ S[M+H]+calculated 316.1114, found 316.1108 (error 1.9 ppm).

This synthetic route provides a better route, allowing for more varied substitution of the C7 position of 4-amino-quinoline, and is demonstrated by the synthesis of 4-amino-2-butyl-7-methoxycarbonylthiazolo [4,5-C ] quinoline 1. This approach overcomes the limitations of the conventional route that requires the formation of 5N-oxide intermediates that are incompatible with the electron withdrawing group at the C7 position. In contrast, it has surprisingly been found that d-catalyzed carbonylation reactions allow for the design and efficient and scalable synthesis of substituted quinoline nucleoside base analogs as well as highly substituted isoquinolines.

Example 11

Synthesis of 4-amino-2-butyl-7-methoxycarbonylthiazolo [4,5-c ] quinoline (1)

Compound 1 was synthesized according to the following reaction conditions.

Example 12

Heterocyclic compounds of the present disclosure

The compounds of Table 2 were produced using the synthetic methods described in examples 1-11 and schemes 1-5.

TABLE 2 Compounds produced according to embodiments of the present disclosure

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Example 13

Assay of test compounds using TLR7/8-NF- κB reporter gene

Human Embryonic Kidney (HEK) cells stably transfected with human TLR-7 or TLR-8 and NF-. Kappa.B-responsive Secreted Embryonic Alkaline Phosphatase (SEAP) genes (HEK-TLR-7 and-8) were purchased from InvivoGen (San Diego, calif.). HEK-TLR7/8 cells were stimulated with 30. Mu.M compound in 96-well plates containing 10% FBS and 0.01% Normocin (InvivoGen) DMEM for 24 hours. mu.L of supernatant from each well was incubated with the Quanti-blue substrate solution (InvivoGen) for 1h at 37℃and absorbance was read at 650nm using a Synergy plate reader (Biotek, winioski, VT).

Example 14

Measurement of proinflammatory cytokines using flow microbead assay

Bone marrow derived dendritic cells (BMDCs) were generated by isolating a single cell suspension of bone marrow from the femur of C57BL/6 mice (6-8 weeks old). With 0.83% NH ₄ Cl、0.1％KHCO ₃ And 0.009% lysed erythrocytes. 500 ten thousand cells were seeded in full RPMI medium (Invitrogen, grand Island, N.Y.) in each well of a 6-well plate, supplemented with mouse 20ng/ml granulocyte-macrophage colony stimulating factor (PeproTech, rocky Hill, N.J.). Approximately 6 days after incubation, BMDC was stimulated with 30. Mu.M compound for 3 days. 25 μl of the supernatant was then removed and tnfα, IL-12p40, IL-1 β and IL-10 were determined using a flow cytometric bead array according to the manufacturer's instructions (BD Bioscience, san Jose, CA). Controls were performed by adding the medium without drug and the vehicle. Flow cytometry was performed on a FACS canto-II (BD Bioscience) and the data was analyzed using Flowjo software (Tree Star, inc. Ashland, OR).

Example 15

Measurement of proinflammatory cytokines using flow microbead assay

Dendritic cells are produced by peripheral blood mononuclear cells as described (Brossart P et al blood 1998; 92:4238-4247). Briefly, CD14 positive monocytes were from healthy human Peripheral Blood Mononuclear Cells (PBMC) obtained via isolation with lymphocyte separation medium (Mediatech, inc, manassas, VA) and after purification with CD14 microbeads from Miltenyi Biotec Inc (Aubun, CA). CD14 positive monocytes (> 95% CD 14) were cultured into immature monocyte-derived dendritic cells (MoDC) by further culturing for 6 days with GM-CSF (100 ng/ml) and IL-4 (100 ng/ml) (R & D, MN).

Approximately 10 ten thousand MoDC cells were plated into 96-well plates and stimulated in triplicate with 5 different concentrations of TLR modulating compound of the present disclosure at concentrations of 0, 0.325, 1.3, 5.2 and 20.8nmol/ml for 48 hours. For immunostaining and flow cytometry analysis, and approximately 48 hours after stimulation with the compounds, cells were stained with anti-HLA-DR, CD11c, CD-86, CD80, CD83, CD8a, CD123 and related isotype controls (eBioscience, san Diego, calif.). Cells were loaded onto FACS-canto II and analyzed with FACSDiva and Flowjo. For flow microbead assay (CBA), supernatants were harvested 48 hours after stimulation with TLR modulating compound. The inflammatory cytokine levels were determined with CBA according to the manufacturer's instructions (BD, san Jose, CA).

Example 16

Evaluation of IL-6 levels following stimulation with compounds of the present disclosure

TLR7 mutant mice and C57BL/6j mice at 8-12 weeks of age were obtained from Jackson Lab (Bar Harbor, main). The TLR7 mutant gene was introduced into 129S1/Sv derived from CJ7 embryonic stem cells. This cell line was backcrossed ten times with C57BL/6 Ncr. TLR7RNA expression was not detected in bone marrow derived macrophages. The homologous TLR7 mutant mice are bred by backcrossing heterologous mutant mice and wild C57BL/6 j.

After squeezing the whole spleen through a 70 μm cell filter and performing a erythrocyte lysis process, single cell suspensions of spleen cells from C57BL/6j or TLR7 mutant mice were isolated. Splenocytes were pulsed in triplicate with complete RPMI-1640 medium (10% heat-inactivated FBS, glutamine, 1% penicillin/streptomycin, 55nmol 2-ME, 10mmol HEPES) containing 2.08nmol/ml or 20.8nmol/ml Imiquimod (IMQ), hydroxy imiquimod (IMQ-OH) or 10 μg/ml CpG 685. Supernatants from the media were harvested 12 hours and 24 hours post-pulse and frozen at-80 ℃ until analysis. According to the manufacturer's instructions, a flow microbead array (BD Biosciences, san Jose, calif.) was used to measure IL-6 levels. Analyses were performed on a FACScanto-II machine using FACSAria II software and further analysis was performed using Flowjo software (Tree Star, inc, ashland, OR). Standard curves and negative controls (PBS) were included for calculation of cytokine concentrations in samples.

Example 17

In vitro binding Capacity screening of TLR7/8 expressing cells

TLR-7 or TLR-8 positive cell lines, such as HEK-Blue TLR cells (invitrogen, san Diego, CA) were used for this screening assay. HEK-Blue TLR cells are engineered HEK293 cells. Stable expression and induction of TLR genes by such cells Type NF-kB-SEAP (secreted embryo alkaline phosphatase) reporter gene. Binding of a ligand (e.g., a compound of the present disclosure) to a TLR in a HEK-Blue cell induces SEAP to produce a Blue color. Screening assays were performed according to the manufacturer's instructions. TLR targeting compounds at concentrations of 20.8nmol/ml or 5.2nmol/ml were added in triplicate to a mixture of 5% CO at 37 ℃ ₂ HEK-Blue-TLR7 or TLR8 cells cultured under conditions. After 24 hours, 5. Mu.l of culture supernatant was mixed with 200. Mu.l of detection medium containing pNPP. After one hour, SEAP activity was read as OD at 650nm using a microplate reader (BioTek Synergy 2, vermont).

Example 18

Detection of inflammatory cytokines in BMDC and spleen cells

Bone marrow cells were harvested from the femur and tibia of C57BL/6j mice. After removal of erythrocytes with potassium ammonium chloride buffer, bone marrow cells were incubated with complete RPMI-1640 medium and 2ng/ml granulocyte macrophage colony stimulating factor (GM-CFS) at 5% CO ₂ And incubated at 37℃for 6 days. During the 6 day incubation period, the medium was changed twice. Single cell suspensions of splenocytes were prepared as described herein. BMDC or splenocytes were stimulated in triplicate with different concentrations of test compound of formula I of 20.8nmol/ml, 5.2nmol/ml, 1.3nmol/ml, 0.325nmol/ml and 0 nmol/ml. 48 hours after stimulation, the supernatant was harvested and frozen at-80 ℃ until analysis. Inflammatory cytokines were subjected to a flow microbead array (CBA, BD Bioscience) according to the manufacturer's instructions. 500 events were collected. All samples were analyzed on a FACScanto-II machine with software and further analysis with Flowjo. Each cytokine included a standard curve and a negative control (PBS) to calculate the concentration of the cytokine in the sample.

Costimulator levels on BMDCs were measured. BMDCs were stained in triplicate with different fluorescently labeled antibodies obtained from eBioscience (San Diego, CA) two days after stimulation with the test compound of formula I. Antibodies included anti-MHC-II (I-A/I-E, clone M5/114.15.2), anti-CD 86 (clone GL 1), anti-CD 80 (clone 16-10A 1), anti-CD 8 alpha (clone 53-6.7), anti-CD 11b (clone M1/70), anti-CD 205 (clone 205 yekta), anti-CD 3 (clone 17A 2), and anti-CD 11c (clone N418). All samples were collected on a FACSCanto II flow cytometer (BD Biosciences, san Jose, CA). 50,000 to 100,000 events were collected. All data were analyzed using Flowjo software (Tree Star, inc, ashland, OR). Gating was based on the CD3-cd11c+ population.

Proliferation of pmel CD8 and IFNγ production after cross presentation. BMDC from C56BL/6j mice and BMDC from pmel mice (containing human gp 100) _25-33 A single cell suspension of splenocytes from H2Db specific receptor T cell receptor transgenic mice, jackson Lab) was prepared as described in this example. BMDCs were pulsed in triplicate with 3.5 μg human gp100 peptide per well (peptide sequence: CALLAVGATKVPRNQDWLGVSRQLRTK, genScript, piscataway, NJ) and test compound of formula I at a concentration of 10.4nmol/ml for 48 hours with the hgp100 peptide control and PBS negative control. BMDCs were washed twice with complete RPMI medium and then co-cultured with pmel CD8 splenocytes, which were CFSE-labeled and isolated from pmel splenocytes using a cd8+ T cell isolation kit (Miltenyi Biotec, auburn, CA) at a DC/CD8 ratio of 1:3. Four days after co-cultivation, the supernatant was harvested and frozen at-80 ℃ until infγ was detected with CBA kit. CBA of ifnγ measurement was performed according to the manufacturer's instructions. The cell pellet was washed and stained with fluorescently labeled antibodies, all obtained from eBioscience. They are anti-CD 3 (clone, 17A 2) and anti-CD 8 alpha (clone 53-6.7). Flow cytometer data were obtained from stained samples on a FACSCanto II flow cytometer and analyzed using Flowjo software. Gating was from the cd3+cd8+ population.

OT-I cells produce IL-2 after stimulation with IMQ-derived novel TLR7 ligands. Single bone marrow cell suspensions from C57BL/6j mice were prepared as previously described in this example. Cells were pulsed in triplicate with a concentration of 20.8nmol/ml of test compound of formula I, with and without 15 μg ovalbumin (Sigma-Aldrich, st. Louis, MO) added per well. Four days later, the cells were washed twice with complete RPMI medium and incubated with isolated OT-I CD8T cells using the cd8+ isolation kit (Miltenyl Biotec, auburn, CA). Four days after co-cultivation, the supernatant was harvested and examined. CBA was performed for IL-2 production according to the specifications of BD Bioscience. Data were collected on a FACSCanto-II flow cytometer and 500 events were collected and analyzed with Flowjo software.

The terms and expressions which have been employed herein are used as terms of description and not of limitation, and there is no intention in the use of such terms and expressions of excluding any equivalents of the features shown and described or portions thereof, but it is recognized that various modifications are possible within the scope of the disclosure claimed. Accordingly, it should be understood that while the present disclosure has been specifically disclosed by some embodiments, exemplary embodiments and optional features, modification and variation of the concepts herein disclosed may be resorted to by those skilled in the art, and that such modifications and variations are considered to be within the scope of this disclosure as defined by the appended claims. The specific embodiments provided herein are examples of useful embodiments of the present disclosure, and it will be apparent to those skilled in the art that the present disclosure may be practiced with numerous variations of the apparatus, apparatus assemblies, method steps set forth in the present specification. It will be apparent to those skilled in the art that the methods and apparatus useful in the present methods may include a wide variety of optional compositions and processing elements and steps.

Claims

1. A compound of formula I:

wherein:

fused ring a is selected from the group consisting of:

R ¹ is H, (C) ₁ -C ₆ ) Alkyl, (C) ₂ -C ₆ ) Alkenyl group (C) ₂ -C ₆ ) Alkynyl, (C) ₁ -C ₆ ) Alkanoyl, (C) ₁ -C ₆ ) Alkoxycarbonyl, (C) ₁ -C ₆ ) Alkanoyloxy, (C) ₃ -C ₆ ) Cycloalkyl, arylRadicals, heteroaryl, heterocyclic rings, NR ^g R ^h Or R is ^m R ⁿ NC (=o) -, any of which (C) _1- C ₆ ) Alkyl, (C) ₂ -C ₆ ) Alkenyl group (C) ₂ -C ₆ ) Alkynyl, (C) ₁ -C ₆ ) Alkanoyl, (C) ₁ -C ₆ ) Alkoxycarbonyl, (C) ₁ -C ₆ ) Alkanoyloxy, (C) ₃ -C ₆ ) Cycloalkyl, aryl, heteroaryl, and heterocycle are optionally substituted with one or more groups independently selected from the group consisting of: halo, cyano, oxo, oxirane, (C) ₃ -C ₈ ) Cycloalkyl, optionally substituted by NR ^u R ^v Substituted alkyl-substituted aryl, carboxy-substituted aryl, heteroaryl, (C) ₁ -C ₆ ) Alkoxy, (C) ₁ -C ₆ ) Alkylthio, OR ^z 、-N(H)S(O) ₂ R ^r 、R ^s C(＝O)O-、-S-R ^w 、-NR ^x R ^y 、(C ₁ -C ₆ ) Alkoxycarbonyl and carboxyl;

R ⁴ Is R ^w S(O) ₂ NR ^f -；

R ^b Is H or X-Y;

R ^e is H, (C) ₁ -C ₆ ) Alkyl or R ^aa ；

R ^f Is H or (C) ₁ -C ₆ ) An alkyl group;

x is a linking group; and is also provided with

Y is a peptide, protein or maleimide;

each R ^u And R is ^v Independently H or (C) ₁ -C ₆ ) An alkyl group; or together with the nitrogen to which they are attached form an aziridinyl, azetidinyl, morpholino, piperazino, pyrrolidino or piperidinyl ring, said ring optionally being capable of being substituted with one or moreC ₁ -C ₆ ) Alkyl substitution;

or a pharmaceutically acceptable salt thereof.

2. A compound of formula I:

wherein:

fused ring a is selected from the group consisting of:

R ¹ is H, (C) ₁ -C ₆ ) Alkyl, (C) ₂ -C ₆ ) Alkenyl group (C) ₂ -C ₆ ) Alkynyl, (C) ₁ -C ₆ ) Alkanoyl, (C) ₁ -C ₆ ) Alkoxycarbonyl, (C) ₁ -C ₆ ) Alkanoyloxy, (C) ₃ -C ₆ ) Cycloalkyl, aryl, heteroaryl,Heterocycle, NR ^g R ^h Or R is ^m R ⁿ NC (=o) -, any of which (C) ₁ -C ₆ ) Alkyl, (C) ₂ -C ₆ ) Alkenyl group (C) ₂ -C ₆ ) Alkynyl, (C) ₁ -C ₆ ) Alkanoyl, (C) ₁ -C ₆ ) Alkoxycarbonyl, (C) ₁ -C ₆ ) Alkanoyloxy, (C) ₃ -C ₆ ) Cycloalkyl, aryl, heteroaryl, and heterocycle are optionally substituted with one or more groups independently selected from the group consisting of: halo, cyano, oxo, oxirane, (C) ₃ -C ₈ ) Cycloalkyl, optionally substituted by NR ^u R ^v Substituted alkyl-substituted aryl, carboxy-substituted aryl, heteroaryl, (C) ₁ -C ₆ ) Alkoxy, (C) ₁ -C ₆ ) Alkylthio, OR ^z 、-N(H)S(O) ₂ R ^r 、R ^s C(＝O)O-、-S-R ^w 、-NR ^x R ^y 、(C ₁ -C ₆ ) Alkoxycarbonyl and carboxyl;

R ⁴ Is at the 2-position by R ^k -C(＝O)-、R ^k -O-C(＝O)-、R ^k -S(O) ₂ -O-、

R ^c R ^d NC(＝O)-、R ^c R ^d NS(O) ₂ -、R ^e C(＝O)N(R ^e ) -or R ^e S(O) ₂ NR ^f Substituted 1-

Ethylene;

R ^b Is H or X-Y;

R ^e is H, (C) ₁ -C ₆ ) Alkyl or R ^aa ；

R ^f Is H or (C) ₁ -C ₆ ) An alkyl group;

R ^k is H, (C) ₁ -C ₆ ) Alkyl, (C) ₂ -C ₆ ) Alkenyl group (C) ₂ -C ₆ ) Alkynyl, (C) ₃ -C ₈ ) Cycloalkyl, trifluoromethyl, aryl or aryl (C) ₁ -C ₆ ) Alkyl groups, each of which (C ₁ -C ₆ ) Alkyl (C)The radicals being optionally substituted by one or more halogen radicals, (C) ₁ -C ₆ ) Alkanoyloxy, (C) ₁ -C ₆ ) Alkoxy, (C) ₃ -C ₈ ) Cycloalkyl substitution;

x is a linking group; and is also provided with

Y is a peptide, protein or maleimide;

or a pharmaceutically acceptable salt thereof.

3. The compound of claim 1 or 2, wherein R ^k Is H, (C) ₁ -C ₆ ) Alkyl, (C) ₂ -C ₆ ) Alkenyl group (C) ₂ -C ₆ ) Alkynyl, (C) ₃ -C ₈ ) Cycloalkyl, trifluoromethyl, aryl or aryl (C) ₁ -C ₆ ) Alkyl groups, each of which (C ₁ -C ₆ ) The alkyl group being optionally substituted by one or more halo groups, (C) ₁ -C ₆ ) Alkoxyacyl group (C) ₁ -C ₆ ) Alkoxy or (C) ₃ -C ₈ ) Cycloalkyl substitution.

4. The compound of any one of claims 1-6, wherein ring a is selected from the group consisting of:

5. the compound of claim 4, wherein R ¹ And R is ³ Each independently is H or optionally substituted (C) ₁ -C ₆ ) An alkyl group.

6. The compound of claim 4 or 5, wherein R ² Is H, (C) ₁ -C ₆ ) Alkyl, (C) ₂ -C ₆ ) Alkenyl group (C) ₂ -C ₆ ) Alkynyl, (C) ₁ -C ₆ ) Alkanoyl, (C) ₁ -C ₆ ) Alkoxycarbonyl, (C) ₁ -C ₆ ) Alkanoyloxy, (C) ₃ -C ₆ ) Cycloalkyl, aryl, heteroaryl, heterocycle, NR ^g R ^h Or R is ^m R ⁿ NC (=o) -, any of which (C) ₁ -C ₆ ) Alkyl, (C) ₂ -C ₆ ) Alkenyl group (C) ₂ -C ₆ ) Alkynyl, (C) ₁ -C ₆ ) Alkanoyl, (C) ₁ -C ₆ ) Alkoxycarbonyl, (C) ₁ -C ₆ ) Alkanoyloxy, (C) ₃ -C ₆ ) Cycloalkyl, aryl, heteroaryl, and heterocycle are optionally substituted with one or more groups independently selected from the group consisting of: hydroxy, halo, -SH, cyano, oxo, oxirane, (C) ₃ -C ₈ ) Cycloalkyl, aryl, heteroaryl, (C) ₁ -C ₆ ) Alkoxy, (C) ₁ -C ₆ ) Alkylthio and NR ^g R ^h 。

7. The compound of any one of claims 1-6, wherein ring a is selected from the group consisting of:

8. the compound of any one of claims 1-7, wherein R ² Is H or optionally substituted (C) ₁ -C ₆ ) An alkyl group.

9. The compound of any one of claims 6-8, wherein ring a is:

10. the compound of any one of claims 6-8, wherein ring a is:

11. the compound of any one of claims 6-8, wherein ring a is:

12. the compound of any one of claims 6-8, wherein ring a is:

13. a compound of formula I:

wherein:

the condensed ring A is:

R ⁴ Is R ^k -C(＝O)-、R ^k -O-C(＝O)-、R ^k -S(O) ₂ -O-、R ^c R ^d NC(＝O)-、

R ^c R ^d NS(O) ₂ -、R ^e C(＝O)N(R ^e )-、R ^e S(O) ₂ NR ^f -, or at the 2-position by R ^k -C(＝O)-、R ^k -O-C(＝O)-、R ^k -S(O) ₂ -O-、R ^c R ^d NC(＝O)-、R ^c R ^d NS(O) ₂ -、R ^e C(＝O)N(R ^e ) -or R ^e S(O) ₂ NR ^f -substituted 1-ethylene;

R ^b Is H or X-Y;

x is a linking group; and is also provided with

Y is a peptide, protein or maleimide.

14. A compound of formula I:

Wherein:

the condensed ring A is:

R ^c R ^d NS(O) ₂ -、R ^e C(＝O)N(R ^e )-、R ^e S(O) ₂ NR ^f -, or at the 2-position by R ^k -C(＝O)-、

R ^k -O-C(＝O)-、R ^k -S(O) ₂ -O-、R ^c R ^d NC(＝O)-、R ^c R ^d NS(O) ₂ -、R ^e C(＝O)N(R ^e )-

Or R is ^e S(O) ₂ NR ^f -substituted 1-ethylene;

R ^b Is H or X-Y;

x is a linking group; and is also provided with

Y is a peptide, protein or maleimide.

15. A compound of formula I:

wherein:

the condensed ring A is:

R ² is H, (C) ₁ -C ₆ ) Alkyl, (C) ₂ -C ₆ ) Alkenyl group (C) ₂ -C ₆ ) Alkynyl, (C) ₁ -C ₆ ) Alkanoyl, (C) ₁ -C ₆ ) Alkoxycarbonyl, (C) ₁ -C ₆ ) Alkanoyloxy, (C) ₃ -C ₆ ) Cycloalkyl, aryl, heteroaryl, heterocycle, NR ^g R ^h Or R is ^m R ⁿ NC (=o) -, any of which (C) ₁ -C ₆ ) Alkyl, (C) ₂ -C ₆ ) Alkenyl group (C) ₂ -C ₆ ) Alkynyl, (C) ₁ -C ₆ ) Alkanoyl, (C) ₁ -C ₆ ) Alkoxycarbonyl, (C) ₁ -C ₆ ) Alkanoyloxy, (C) ₃ -C ₆ ) Cycloalkyl, aryl, heteroaryl, and heterocycle are optionally substituted with one or more groups independently selected from the group consisting of: hydroxy, halo, -SH, cyano, oxo, oxirane 、(C ₃ -C ₈ ) Cycloalkyl, aryl, heteroaryl, (C) ₁ -C ₆ ) Alkoxy, (C) ₁ -C ₆ ) Alkylthio and NR ^g R ^h ；

Or R is ^e S(O) ₂ NR ^f -substituted 1-ethylene;

R ^b Is H or X-Y;

x is a linking group; and is also provided with

Y is a peptide, protein or maleimide.

16. A compound of formula I:

wherein:

the condensed ring A is:

R ¹ is H, (C) ₁ -C ₆ ) Alkyl, (C) ₂ -C ₆ ) Alkenyl group (C) ₂ -C ₆ ) Alkynyl, (C) ₁ -C ₆ ) Alkanoyl, (C) ₁ -C ₆ ) Alkoxycarbonyl, (C) ₁ -C ₆ ) Alkanoyloxy, (C) ₃ -C ₆ ) Cycloalkyl, aryl, heteroaryl, heterocycle, NR ^g R ^h Or R is ^m R ⁿ NC (=o) -, any of whichC ₁ -C ₆ ) Alkyl, (C) ₂ -C ₆ ) Alkenyl group (C) ₂ -C ₆ ) Alkynyl, (C) ₁ -C ₆ ) Alkanoyl, (C) ₁ -C ₆ ) Incubating carbonyl group (C) ₁ -C ₆ ) Alkanoyloxy, (C) ₃ -C ₆ ) Cycloalkyl, aryl, heteroaryl, and heterocycle are optionally substituted with one or more groups independently selected from the group consisting of: halo, cyano, oxo, oxirane, (C) ₃ -C ₈ ) Cycloalkyl, aryl optionally substituted by alkyl, aryl substituted by carboxyl, heteroaryl, (C) ₁ -C ₆ ) Alkoxy, (C) ₁ -C ₆ ) Alkylthio, OR ^z 、-N(H)S(O) ₂ R ^r 、R ^s C(＝O)O-、-S-R ^w 、-NR ^x R ^y 、(C ₁ -C ₆ ) Alkoxycarbonyl and carboxyl, said alkyl being substituted by NR ^u R ^v Substitution;

Or R is ^e S(O) ₂ NR ^f -substituted 1-ethylene;

R ^a is H, (C) ₁ -C ₆ ) Alkyl, (C) ₂ -C ₆ ) Alkenyl group (C) ₂ -C ₆ ) Alkynyl, (C) ₁ -C ₆ ) Alkanoyl, (C) ₁ -C ₆ ) Alkoxycarbonyl, (C) ₃ -C ₆ ) Cycloalkyl, aryl or heteroaryl, any of whichWhat (C) ₁ -C ₆ ) Alkyl, (C) ₂ -C ₆ ) Alkenyl group (C) ₂ -C ₆ ) Alkynyl, (C) ₁ -C ₆ ) Alkanoyl, (C) ₁ -C ₆ ) Alkoxycarbonyl, (C) ₃ -C ₆ ) Cycloalkyl, aryl, and heteroaryl are optionally substituted with one or more groups independently selected from the group consisting of: hydroxy, halo, -SH, cyano, oxirane, (C) ₃ -C ₈ ) Cycloalkyl, aryl, heteroaryl, (C) ₁ -C ₆ ) Alkoxy, (C) ₁ -C ₆ ) Alkylthio and NR ^g R ^h ；

R ^b Is H or X-Y;

R ^e is H, (C) ₁ -C ₆ ) Alkyl or R ^aa ；

R ^f Is H or (C) ₁ -C ₆ ) An alkyl group;

x is a linking group; and is also provided with

Y is a peptide, protein or maleimide;

R ^r Is H, arylOr optionally substituted by halogen or aryl (C ₁ -C ₁₀ ) An alkyl group;

R ^aa Is aryl optionally substituted with one or more groups independently selected from the group consisting of: hydroxy, halo, nitro, cyano, (C) ₁ -C ₆ ) Alkoxy and optionally substituted with one or more halo (C ₁ -C ₆ ) An alkyl group;

or a pharmaceutically acceptable salt thereof.

17. A compound of formula II:

Or a pharmaceutically acceptable salt thereof,

wherein:

R ² is H,(C ₁ -C ₆ ) Alkyl, (C) ₂ -C ₆ ) Alkenyl group (C) ₂ -C ₆ ) Alkynyl, (C) ₁ -C ₆ ) Alkanoyl, (C) ₁ -C ₆ ) Alkoxycarbonyl, (C) ₁ -C ₆ ) Alkanoyloxy, (C) ₃ -C ₆ ) Cycloalkyl, aryl, heteroaryl, heterocycle, NR ^g R ^h Or R is ^m R ⁿ NC (=o) -, any of which (C) ₁ -C ₆ ) Alkyl, (C) ₂ -C ₆ ) Alkenyl group (C) ₂ -C ₆ ) Alkynyl, (C) ₁ -C ₆ ) Alkanoyl, (C) ₁ -C ₆ ) Alkoxycarbonyl, (C) ₁ -C ₆ ) Alkanoyloxy, (C) ₃ -C ₆ ) Cycloalkyl, aryl, heteroaryl, and heterocycle are optionally substituted with one or more groups independently selected from the group consisting of: hydroxy, halo, -SH, cyano, oxo, oxirane, (C) ₃ -C ₈ ) Cycloalkyl, aryl, heteroaryl, (C) ₁ -C ₆ ) Alkoxy, (C) ₁ -C ₆ ) Alkylthio and NR ^g R ^h ；

R ⁴ Is R ^k -C (=o) -or R ^k -O-C(＝O)-；

R ^a Is H or optionally substituted (C) ₁ -C ₆ ) An alkyl group;

R ^b is H or X-Y;

x is a linking group; and is also provided with

Y is a peptide, protein or maleimide.

18. The compound of claim 17, wherein R ² Is H or optionally substituted (C) ₁ -C ₆ ) An alkyl group.

19. The compound of any one of claims 17 or 18, wherein R ^k Is H or optionally substituted (C) ₁ -C ₆ ) An alkyl group.

20. The compound of any one of claims 17-19, having the formula:

21. a compound of formula III or formula IV:

or a pharmaceutically acceptable salt thereof,

wherein:

R ⁴ Is R ^k -C(＝O)-、R ^k -O-C (=o) -or R ^k -S(O) ₂ -O-；

R ^b Is H or X-Y;

x is a linking group; and is also provided with

Y is a peptide, protein or maleimide.

22. The compound of claim 21, wherein R ¹ And R is ² Independently is H or can optionally be covered (C) ₁ -C ₃ ) Alkyl, substituted or unsubstituted (C ₃ -C ₈ ) Cycloalkyl or substituted or unsubstituted aryl substituted (C ₁ -C ₆ ) An alkyl group.

23. The compound of claim 21 or 22, wherein R ⁴ Is R ^k -O-C (=o) -or R ^k -S(O) ₂ -O-。

24. The compound of claim 21 or 22, selected from the group consisting of:

25. the compound of any one of claims 1-19 or 21-23, wherein X is (C ₁ -C ₆ ) Alkyl, (C) ₂ -C ₆ ) Alkenyl or (C) ₁ -C ₆ ) Alkynyl, said (C ₁ -C ₆ ) Alkyl, (C) ₂ -C ₆ ) Alkenyl or (C) ₁ -C ₆ ) Alkynyl groups are optionally substituted with oxo groups.

26. The compound of any one of claims 1-19 or 21-23, wherein X is selected from the group consisting of:

and wherein n is 2, 3, 4, 5 or 6.

27. The compound of any one of claims 1-19, 21-23, or 25-26, wherein Y is maleimide.

28. The compound of any one of claims 1-19, 21-23, or 25-26, wherein Y is an antigen.

29. The compound of claim 28, wherein the antigen is associated with a bacterium or virus, and wherein the virus is selected from the group consisting of influenza virus, HIV, and HCV.

30. The compound of any one of claims 1-19, 21-23, or 25-26, wherein Y is an antigen associated with a tumor cell or tumor cell lysate.

31. The compound of any one of claims 28-30, wherein Y is an antigen comprising a peptide sequence comprising cysteine, lysine, or both.

32. The compound of any one of claims 1-31, wherein the compound is any one of compounds 1 or 22-26.

33. A compound of formula I:

Wherein:

fused ring a is selected from the group consisting of:

R ² is H, (C) ₁ -C ₆ ) Alkyl, (C) ₂ -C ₆ ) Alkenyl group (C) ₂ -C ₆ ) Alkynyl, (C) ₁ -C ₆ ) Alkanoyl, (C) ₁ -C ₆ ) Alkoxycarbonyl, (C) ₁ -C ₆ ) Alkanoyloxy, (C) ₃ -C ₆ ) Cycloalkyl, aryl, heteroaryl, heterocycle, NR ^g R ^h Or R is ^m R ⁿ NC (=o) -, any of which (C) ₁ -C ₆ ) Alkyl, (C) ₂ -C ₆ ) Alkenyl group (C) ₂ -C ₆ ) Alkynyl, (C) ₁ -C ₆ ) Alkanoyl, (C) ₁ -C ₆ ) Alkoxycarbonyl, (C) ₁ -C ₆ ) Alkanoyloxy, (C) ₃ -C ₆ ) Cycloalkyl, aryl, heteroaryl and heterocycle are optionally substituted with one or more groups independently selected from the group consisting of Group substitution: hydroxy, halo, -SH, cyano, oxo, oxirane, (C) ₃ -C ₈ ) Cycloalkyl, aryl, heteroaryl, (C) ₁ -C ₆ ) Alkoxy, (C) ₁ -C ₆ ) Alkylthio and NR ^g R ^h ；

Or R is ^e S(O) ₂ NR ^f -substituted 1-ethylAn alkene;

R ^b Is H or X-Y;

R ^e is H, (C) ₁ -C ₆ ) Alkyl or R ^aa ；

R ^f Is H or (C) ₁ -C ₆ ) An alkyl group;

each R ^g And R is ^h Independently H, (C) ₁ -C ₆ ) Alkyl, (C) ₂ -C ₆ ) Alkenyl group (C) ₃ -C ₈ ) Cycloalkyl, (C) ₃ -C ₈ ) Cycloalkyl (C) ₁ -C ₆ ) Alkyl, aryl (C) ₁ -C ₆ ) An alkyl group;or together with the nitrogen to which they are attached form an aziridinyl, azetidinyl, morpholino, piperazino, pyrrolidino or piperidinyl ring, said ring being optionally substituted with one or more (C ₁ -C ₆ ) Alkyl substitution;

x is a linking group; and is also provided with

Y is a peptide, protein or maleimide;

each R ^p And R is ^q Independently H orC ₁ -C ₆ ) An alkyl group; or together with the nitrogen to which they are attached form an aziridinyl, azetidinyl, morpholino, piperazino, pyrrolidino or piperidinyl ring, said ring being optionally substituted with one or more (C ₁ -C ₆ ) Alkyl substitution; and is also provided with

or a pharmaceutically acceptable salt thereof.

34. A compound of formula I:

wherein:

fused ring a is selected from the group consisting of:

R ⁴ Is R ^k -C(＝O)-、R ^k -O-C(＝O)-、R ^k -S(O) ₂ -O-、R ^c R ^d NC(＝O)-、R ^c

R ^d NS(O) ₂ -、R ^e S(O) ₂ NR ^f -, or at the 2-position by R ^k -C(＝O)-、R ^k -O-C(＝O)-、R ^k -S(O) ₂ -O-、R ^c R ^d NC(＝O)-、R ^c R ^d NS(O) ₂ -、R ^e C(＝O)N(R ^e ) -or R ^e S(O) ₂ NR ^f -substituted 1-ethylene;

R ^b Is H or X-Y;

R ^e is H, (C) ₁ -C ₆ ) Alkyl or R ^aa ；

R ^f Is H or (C) ₁ -C ₆ ) An alkyl group;

x is a linking group; and is also provided with

Y is a peptide, protein or maleimide;

or a pharmaceutically acceptable salt thereof.

35. The compound of any one of claims 1-6, 16, 21, 23, 25-31, or 33-34, wherein R ¹ Is H, (C) ₁ -C ₆ ) Alkyl, (C) ₂ -C ₆ ) Alkenyl group (C) ₂ -C ₆ ) Alkynyl, (C) ₁ -C ₆ ) Alkanoyl, (C) ₁ -C ₆ ) Alkoxycarbonyl or-NR ^g R ^h Any of (C) ₁ -C ₆ ) Alkyl, (C) ₂ -C ₆ ) Alkenyl group (C) ₂ -C ₆ ) Alkynyl, (C) ₁ -C ₆ ) Alkanoyl (C) ₁ -C ₆ ) The alkoxycarbonyl group is optionally substituted with one or more groups independently selected from the group consisting of: halo, cyano, oxo, oxirane, (C) ₃ -C ₈ ) Cycloalkyl, optionally substituted by NR ^u R ^v Substituted alkyl-substituted aryl, carboxy-substituted aryl, heteroaryl, (C) ₁ -C ₆ ) Alkoxy, (C) ₁ -C ₆ ) Alkylthio, OR ^z 、-N(H)S(O) ₂ R ^r 、R ^s C(＝O)O-、-S-R ^w 、-NR ^x R ^y 、(C ₁ -C ₆ ) Alkoxycarbonyl and carboxyl.

36. The compound of any one of claims 1 to 6, 16, 21, 23, 25-31 or 33-35, wherein R ¹ Is H, (C) ₁ -C ₆ ) Alkyl, (C) ₂ -C ₆ ) Alkenyl group (C) ₂ -C ₆ ) Alkynyl, (C) ₁ -C ₆ ) Alkanoyl, (C) ₁ -C ₆ ) Alkoxycarbonyl or-NR ^g R ^h Any of (C) ₁ -C ₆ ) Alkyl, (C) ₂ -C ₆ ) Alkenyl group (C) ₂ -C ₆ ) Alkynyl, (C) ₁ -C ₆ ) Alkanoyl (C) ₁ -C ₆ ) Alkoxycarbonyl groups optionally being independently selected from one or more halo groupsCyano, oxo, oxirane, (C) ₃ -C ₈ ) Cycloalkyl groups.

37. The compound of any one of claims 1 to 6, 16, 21, 23, 25-31 or 33-36, wherein R ¹ Is H, (C) ₁ -C ₆ ) Alkyl, (C) ₂ -C ₆ ) Alkenyl group (C) ₂ -C ₆ ) Alkynyl or-NR ^g R ^h Any of (C) ₁ -C ₆ ) Alkyl, (C) ₂ -C ₆ ) Alkenyl group sum (C) ₂ -C ₆ ) Alkynyl groups are optionally substituted with one or more groups independently selected from halo, cyano, oxo, oxirane, (C) ₃ -C ₈ ) Cycloalkyl groups.

38. The compound of any one of claims 1-6, 16, 21, 23, 25-31, or 33-37, wherein R ¹ Is H or optionally substituted with one or more groups independently selected from the group consisting of halo and cyano (C ₁ -C ₆ ) An alkyl group.

39. The compound of any one of claims 1-6, 16, 21, 23, 25-31, or 33-38, wherein R ¹ Is H or (C) ₁ -C ₆ ) An alkyl group.

40. The compound of any one of claims 1-6, 16, 21, 23, 25-31, or 33-39, wherein R ¹ Is H.

41. The compound of any one of claims 1-6, 16, 21, 23, 25-31, or 33-39, wherein R ¹ Is not H.

42. The compound of any one of claims 1-5, 7, 9-17, 19, 21, 23, 25-31, or 33-41, wherein R ² Is H, (C) ₁ -C ₆ ) Alkyl, (C) ₂ -C ₆ ) Alkenyl group (C) ₂ -C ₆ ) Alkynyl, (C) ₁ -C ₆ ) Alkyl (C)Acyl group (C) ₁ -C ₆ ) Alkoxycarbonyl or-NR ^g R ^h Any of (C) ₁ -C ₆ ) Alkyl, (C) ₂ -C ₆ ) Alkenyl group (C) ₂ -C ₆ ) Alkynyl, (C) ₁ -C ₆ ) Alkanoyl (C) ₁ -C ₆ ) The alkoxycarbonyl group is optionally substituted with one or more groups independently selected from the group consisting of: hydroxy, halo, -SH, cyano, oxo, oxirane, (C) ₃ -C ₈ ) Cycloalkyl, aryl, heteroaryl, (C) ₁ -C ₆ ) Alkoxy, (C) ₁ -C ₆ ) Alkylthio and NR ^g R ^h 。

43. The compound of any one of claims 1-5, 7, 9-17, 19, 21, 23, 25-31, or 33-42, wherein R ² Is H, (C) ₁ -C ₆ ) Alkyl, (C) ₂ -C ₆ ) Alkenyl group (C) ₂ -C ₆ ) Alkynyl, (C) ₁ -C ₆ ) Alkanoyl, (C) ₁ -C ₆ ) Alkoxycarbonyl or-NR ^g R ^h Any of (C) ₁ -C ₆ ) Alkyl, (C) ₂ -C ₆ ) Alkenyl group (C) ₂ -C ₆ ) Alkynyl, (C) ₁ -C ₆ ) Alkanoyl (C) ₁ -C ₆ ) Alkoxycarbonyl groups optionally being independently selected from halo, -SH, cyano, (C) ₃ -C ₈ ) Cycloalkyl and NR ^g R ^h The groups of the group are substituted.

44. The compound of any one of claims 1-5, 7, 9-17, 19, 21, 23, 25-31, or 33-43, wherein R ² Is H, (C) ₁ -C ₆ ) Alkyl, (C) ₂ -C ₆ ) Alkenyl or (C) ₂ -C ₆ ) Alkynyl, any of which (C ₁ -C ₆ ) Alkyl, (C) ₂ -C ₆ ) Alkenyl group sum (C) ₂ -C ₆ ) Alkynyl groups are optionally substituted with one or more groups independently selected from halo, -SH, cyano, (C) ₃ -C ₈ ) Cycloalkyl and NR ^g R ^h The groups of the group are substituted.

45. The compound of any one of claims 1-5, 7, 9-17, 19, 21, 23, 25-31, or 33-44, wherein R ² Is H, (C) ₁ -C ₆ ) Alkyl, (C) ₂ -C ₆ ) Alkenyl or (C) ₂ -C ₆ ) Alkynyl, any of which (C ₁ -C ₆ ) Alkyl, (C) ₂ -C ₆ ) Alkenyl group sum (C) ₂ -C ₆ ) Alkynyl groups are optionally substituted with one or more groups independently selected from halo, -SH, cyano, (C) ₃ -C ₈ ) Cycloalkyl and NR ^g R ^h The groups of the group are substituted.

46. The compound of any one of claims 1-5, 7, 9-17, 19, 21, 23, 25-31, or 33-45, wherein R ² Is H or is optionally substituted with one or more groups independently selected from halo, -SH, cyano and NR ^g R ^h Group-substituted (C) ₁ -C ₆ ) An alkyl group.

47. The compound of any one of claims 1-5, 7, 9-17, 19, 21, 23, 25-31, or 33-46, wherein R ² Is H or (C) ₁ -C ₆ ) An alkyl group.

48. The compound of any one of claims 1-5, 7, 9-17, 19, 21, 23, 25-31, or 33-47, wherein R ² Is H.

49. The compound of any one of claims 1-5, 7, 9-17, 19, 21, 23, 25-31, or 33-47, wherein R ² Is not H.

50. The compound of any one of claims 1-4, 6, 16, 25-31, or 33-49, wherein R ³ Is H, halo, hydroxy, (C) ₁ -C ₆ ) Alkyl, (C) ₂ -C ₆ ) Alkenyl group (C) ₂ -C ₆ ) Alkynyl, (C) ₁ -C ₆ ) Alkanoyl, (C) ₁ -C ₆ ) AlkoxycarbonylRadicals or NR ^g R ^h Any of (C) ₁ -C ₆ ) Alkyl, (C) ₂ -C ₆ ) Alkenyl group (C) ₂ -C ₆ ) Alkynyl, (C) ₁ -C ₆ ) Alkanoyl (C) ₁ -C ₆ ) The alkoxycarbonyl group is optionally substituted with one or more groups independently selected from the group consisting of: hydroxy, halo, -SH, cyano, oxo, oxirane, (C) ₃ -C ₈ ) Cycloalkyl, aryl, heteroaryl, (C) ₁ -C ₆ ) Alkoxy, (C) ₁ -C ₆ ) Alkylthio and NR ^g R ^h 。

51. The compound of any one of claims 1-4, 6, 16, 25-31, or 33-50, wherein R ³ Is H, halo, hydroxy, (C) ₁ -C ₆ ) Alkyl, (C) ₂ -C ₆ ) Alkenyl group (C) ₂ -C ₆ ) Alkynyl, (C) ₁ -C ₆ ) Alkanoyl, (C) ₁ -C ₆ ) Alkoxycarbonyl or NR ^g R ^h Any of (C) ₁ -C ₆ ) Alkyl, (C) ₂ -C ₆ ) Alkenyl group (C) ₂ -C ₆ ) Alkynyl, (C) ₁ -C ₆ ) Alkanoyl (C) ₁ -C ₆ ) Alkoxycarbonyl groups optionally being independently selected from halo, -SH, cyano, (C) ₃ -C ₈ ) Cycloalkyl and NR ^g R ^h The groups of the group are substituted.

52. The compound of any one of claims 1-4, 6, 16, 25-31, or 33-51, wherein R ³ Is H, (C) ₁ -C ₆ ) Alkyl, (C) ₂ -C ₆ ) Alkenyl group (C) ₂ -C ₆ ) Alkynyl or NR ^g R ^h Any of (C) ₁ -C ₆ ) Alkyl, (C) ₂ -C ₆ ) Alkenyl group sum (C) ₂ -C ₆ ) Alkynyl groups are optionally substituted with one or more groups independently selected from halo, -SH, cyano, (C) ₃ -C ₈ ) Cycloalkyl and NR ^g R ^h The groups of the group are substituted.

53. The compound of any one of claims 1-4, 6, 16, 25-31, or 33-52, wherein R ³ Is H or is optionally substituted with one or more substituents independently selected from halo, -SH, cyano, (C) ₃ -C ₈ ) Cycloalkyl and NR ^g R ^h Group-substituted (C) ₁ -C ₆ ) An alkyl group.

54. The compound of any one of claims 1-4, 6, 16, 25-31, or 33-53, wherein R ³ Is H or (C) ₁ -C ₆ ) An alkyl group.

55. The compound of any one of claims 1-4, 6, 16, 25-31, or 33-54, wherein R ³ Is H.

56. The compound of any one of claims 1-4, 6, 16, 25-31, or 33-54, wherein R ³ Is not H.

57. The compound of any one of claims 13-16, 21-22, 25-31, or 33-56, wherein R ⁴ Is R ^k -C(＝O)-、R ^k -O-C(＝O)-、R ^k -S(O) ₂ -O-、R ^c R ^d NS(O) ₂ -、R ^e S(O) ₂ NR ^f -, or at the 2-position by R ^k -C(＝O)-、R ^k -O-C(＝O)-、R ^k -S(O) ₂ -O-、R ^c R ^d NS(O) ₂ -or R ^e S(O) ₂ NR ^f -substituted 1-ethylene.

58. The compound of any one of claims 13-16, 21-22, 25-31, or 33-57, wherein R ⁴ Is R ^k -C(＝O)-、R ^k -O-C (=o) -, or at the 2-position by R ^k -C(＝O)-、R ^k -O-C (=o) -substituted 1-ethylene.

59. The compound of any one of claims 13-16, 21-22, 25-31, or 33-57, whichR in (B) ⁴ Is R ^k -S(O) ₂ -O-、R ^c R ^d NS(O) ₂ -、R ^e S(O) ₂ NR ^f -, or at the 2-position by R ^k -S(O) ₂ -O-、R ^c R ^d NS(O) ₂ -or R ^e S(O) ₂ NR ^f -substituted 1-ethylene.

60. The compound of any one of claims 13-16, 21-22, 25-31, or 33-58, wherein R ⁴ Is R ^k -C (=o) -or R ^k -O-C(＝O)-。

61. The compound of any one of claims 1-19, 25-31, or 33-60, wherein the compound of formula I comprises a structure of formula (Ia) or formula (Ib):

62. the compound of any one of claims 1-19, 25-31, or 33-61, wherein the compound of formula I comprises a structure of formula (Ia):

63. the compound of any one of claims 1-19, 21-23, 25-31, or 33-62, wherein R ^a Is H, (C) ₁ -C ₆ ) Alkyl, (C) ₂ -C ₆ ) Alkenyl or (C) ₂ -C ₆ ) Alkynyl, any of which (C ₁ -C ₆ ) Alkyl, (C) ₂ -C ₆ ) Alkenyl or (C) ₂ -C ₆ ) Alkynyl groups are optionally substituted with one or more groups independently selected from the group consisting of: halo, -SH, cyano, oxirane, (C) ₃ -C ₈ ) Cycloalkyl, (C) ₁ -C ₆ ) Alkoxy, (C) ₁ -C ₆ ) Alkylthio and NR ^g R ^h 。

64. The compound of any one of claims 1-19, 21-23, 25-31, or 33-63, wherein R ^a Is H, (C) ₁ -C ₆ ) Alkyl, (C) ₂ -C ₆ ) Alkenyl or (C) ₂ -C ₆ ) Alkynyl, any of which (C ₁ -C ₆ ) Alkyl, (C) ₂ -C ₆ ) Alkenyl or (C) ₂ -C ₆ ) Alkynyl is optionally substituted with one or more groups independently selected from halo, -SH, cyano and NR ^g R ^h The groups of the group are substituted.

65. The compound of any one of claims 1-19, 21-23, 25-31, or 33-64, wherein R ^a Is H or is optionally substituted with one or more substituents independently selected from halo, -SH, cyano and NR ^g R ^h Group-substituted (C) ₁ -C ₆ ) An alkyl group.

66. The compound of any one of claims 1-19, 21-23, 25-31, or 33-65, wherein R ^a Is H or (C) ₁ -C ₆ ) An alkyl group.

67. The compound of any one of claims 1-19, 21-23, 25-31, or 33-66, wherein R ^a Is H.

68. The compound of any one of claims 1-19, 21-23, 25-31, or 33-66, wherein R ^a Is not H.

69. The compound of any one of claims 1-3, 25-31, or 33-68, wherein the fused ring a is selected from the group consisting of:

70. the compound of any one of claims 1-3, 25-31, or 33-69, wherein the fused ring a is selected from the group consisting of:

71. The compound of any one of claims 1-3, 25-31, or 33-70, wherein the fused ring a is selected from the group consisting of:

72. the compound of any one of claims 33-68, wherein the fused ring a is

73. The compound of any one of claims 33-68, wherein the fused ring a is selected from the group consisting of:

74. the compound of any one of claims 33-68 or 73, wherein the fused ring a is selected from the group consisting of:/>

75. the compound of any one of claims 33-68 or 73, wherein the fused ring a is

76. A pharmaceutical composition comprising a compound according to any one of claims 1-75 and a pharmaceutically acceptable diluent or carrier.

77. A method of treating a pathological condition in an animal comprising administering to the animal a compound according to any one of claims 1-75, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition according to claim 76.

78. A method of stimulating an immune response in an animal comprising administering to the animal a compound according to any one of claims 1-75, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition according to claim 76.

79. A method of treating cancer in an animal comprising administering to the animal a compound according to any one of claims 1-75, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition according to claim 76.

80. A method of synthesizing the compound of any one of claims 1-75, the method comprising performing a reaction according to any one of reaction schemes 1-5.