TLR7 / 8 Antagonists and Their Uses

Abstract

Compounds useful as TLR7 / 8 antagonists and pharmaceutically acceptable compositions thereof are provided. The present invention provides a compound represented by formula I.

Description

[Technical Field]

[0001] Related Applications This application claims the benefit of U.S. Provisional Application No. 62 / 533,820, filed July 18, 2017, the contents of which are incorporated by reference in their entirety.

[0002] FIELD OF THE INVENTION The present invention provides compounds of formula (I) as Toll-like receptor 7 / 8 (TLR7 / 8) antagonists and their use in the treatment of immune disorders and other diseases associated with TLR7 / 8 overexpression. [Background technology]

[0003] Background of the Invention Toll-like receptors (TLRs), currently comprising a gene family of 10 receptors with different specificities, are part of the cellular pathogen pattern recognition system that has evolved to defend against various infectious diseases (bacterial, viral, and fungal). TLR activation leads to cytokine responses, including the release of interferon and the activation of specific immune cells. The functional expression of selected TLRs in tissues varies significantly. Some receptors, such as TLR4 (stimulated by E. coli lipopolysaccharide (LPS)), are located on the cell surface, e.g., on epithelial cells, while TLRs 3, 7, 8, and 9 are located on the endosomal membrane in specific immune cells. All of the latter are activated by nucleic acids, but they recognize various types of nucleic acids. For example, TLR9 is activated by single-stranded DNA containing CpG sequences, TLRs 7 and 8 are activated by single-stranded RNA, and TLR3 is activated by double-stranded RNA.

[0004] TLRs are involved in various autoimmune and inflammatory diseases, with the most obvious example being the role played by TLR7 in the pathogenesis of systemic lupus erythematosus (Barrat and Coffman, Immunol Rev, 223:271-283, 2008). In addition, TLR8 polymorphisms are associated with rheumatoid arthritis (Enevold et al., J Rheumatol, 37:905-10, 2010). Although various inhibitors of TLR7, TLR8 and TLR9 have been described, additional TLR inhibitors are desired. In particular, polynucleotides having inhibitory motifs for one or more of TLR7, TLR8 and TLR9 are needed to strictly inhibit immune responses in subjects (e.g., patients with autoimmune diseases or inflammatory disorders).

[0005] In recent years, intense efforts have been made worldwide to harness the potent immune activation induced by TLR7, 8, or 9 agonists for cancer treatment. However, cancer immunotherapy has a long history of failure. However, in recent years, our understanding of cancer immunosurveillance and the resulting functions of immune cell subsets has dramatically improved. TLR7 or TLR9 agonists are in clinical development for cancer monotherapy or combination therapy, or as vaccine adjuvants. The TLR agonist approach for cancer immunotherapy differs from previous efforts using cytokines, interferons, or monovalent vaccinations, for example. TLR agonist-mediated immune activation is multifaceted, mediated by specific immune cells (primarily dendritic cells and B cells, followed by other cells), resulting in innate and adaptive immune responses. Furthermore, not only one type of interferon is induced, but rather many different isoforms are induced simultaneously, including type I (alpha and beta) and type II (gamma and NK cells) (indirectly). Summary of the Invention

[0006] SUMMARY OF THE INVENTION In one aspect, the present invention provides a compound of formula (I): [ka] and pharmaceutically acceptable derivatives, solvates, salts, hydrates and stereoisomers thereof.

[0007] In another aspect, the present invention provides compounds of formula (I) that are dual antagonists of TLR7 and TLR8. In another aspect, the present invention provides compounds of formula (I) that are suitable for the treatment and / or prevention of disorders related to TLR7 / 8. In another aspect, the present invention provides compounds capable of modulating, in particular inhibiting, the activity or function of TLR7 / 8 in mammalian, in particular human, pathological conditions.

[0008] According to another aspect of the present invention, there is provided a method for the treatment and / or prevention of an autoimmune disorder.

[0009] According to another aspect, the present invention provides compounds of formula (I) that are selective for TLR7 or TLR8.

[0010] According to another aspect, the present invention provides compounds of formula (I) that are selective for TLR7 and TLR8. DETAILED DESCRIPTION OF THE INVENTION

[0011] DETAILED DESCRIPTION OF CERTAIN EMBODIMENTS 1. General Description of the Compounds of the Invention In some aspects, the present invention provides antagonists of TLR7 / 8. In some embodiments, such compounds include compounds represented by the formulas described herein, or pharmaceutically acceptable salts thereof, wherein each variable is as defined and described herein.

[0012] 2. Compounds and Definitions The compounds of the present invention include those generally described above and are further described by the classes, subclasses, and species disclosed herein. As used herein, the following definitions may apply unless otherwise indicated. For purposes of the present invention, chemical elements are defined as defined in the Elements, CAS version, Handbook of Chemistry and Physics, 75 th In addition, the general principles of organic chemistry are identified in "Organic Chemistry", Thomas Sorrell, University Science Books, Sausalito: 1999, and "March's Advanced Organic Chemistry", 5 th Ed., Ed.: Smith, MB and March, J., John Wiley & Sons, New York: 2001, the contents of which are incorporated herein by reference in their entireties.

[0013] The terms "aliphatic" or "aliphatic group," as used herein, refer to a straight-chain (i.e., unbranched) or branched, substituted or unsubstituted hydrocarbon chain that is fully saturated or contains one or more units of unsaturation, or a monocyclic or bicyclic hydrocarbon that is fully saturated or contains one or more units of unsaturation, but which is not aromatic (also referred to herein as "carbocyclic," "alicyclic," or "cycloalkyl") and has a single point of attachment to the rest of the molecule. Unless so specified, an aliphatic group contains 1-6 aliphatic carbon atoms. In some embodiments, an aliphatic group contains 1-5 aliphatic carbon atoms. In other embodiments, an aliphatic group contains 1-4 aliphatic carbon atoms. In still other embodiments, an aliphatic group contains 1-3 aliphatic carbon atoms, and in still other embodiments, an aliphatic group contains 1-2 aliphatic carbon atoms. In some embodiments, "alicyclic" (or "carbocycle" or "cycloalkyl") refers to a monocyclic C3-C6 hydrocarbon that is fully saturated or contains one or more units of unsaturation, but which is not aromatic and has a single point of attachment to the rest of the molecule. Exemplary aliphatic groups are linear or branched, substituted or unsubstituted C1-C8 alkyl groups, C2-C8 alkenyl groups, C2-C8 alkynyl groups, and hybrids thereof, such as (cycloalkyl)alkyl, (cycloalkenyl)alkyl, or (cycloalkyl)alkenyl.

[0014] The term "lower alkyl" refers to a C 1~4 It refers to a straight or branched chain alkyl group. Exemplary lower alkyl groups are methyl, ethyl, propyl, isopropyl, butyl, isobutyl, and tert-butyl.

[0015] The term "lower haloalkyl" refers to a C alkyl group substituted with one or more halogen atoms. 1~4 It refers to a straight or branched chain alkyl group.

[0016] The term "heteroatom" refers to one or more of oxygen, sulfur, nitrogen, or phosphorus (including any oxidized form of nitrogen, sulfur, or phosphorus; the quaternized form of any basic nitrogen; or a substitutable nitrogen of a heterocycle, such as N (as in 3,4-dihydro-2H-pyrrolyl), NH (as in pyrrolidinyl), or NR + (including those found in N-substituted pyrrolidinyls).

[0017] The term "unsaturated," as used herein, means that a moiety has one or more units of unsaturation.

[0018] As used herein, the term "divalent C 1~8 (or C 1~6 ) saturated or unsaturated, linear (straight chain) or branched, hydrocarbon chain" refers to divalent alkylene chains, alkenylene chains, and alkynylene chains, straight chains or branched chains as defined herein.

[0019] The term "alkylene" refers to a divalent alkyl group. An "alkylene chain" is a polymethylene group, i.e., -(CH) n -, where n is a positive integer, preferably 1 to 6, 1 to 4, 1 to 3, 1 to 2, or 2 to 3. A substituted alkylene chain is a polymethylene group in which one or more methylene hydrogen atoms are replaced with a substituent. Suitable substituents include those described below for substituted aliphatic groups.

[0020] The term "alkenylene" refers to a divalent alkenyl group. A substituted alkenylene chain is a polymethylene group containing at least one double bond in which one or more hydrogen atoms are replaced with a substituent. Suitable substituents include those described below for substituted aliphatic groups.

[0021] The term "halogen" means F, Cl, Br, or I.

[0022] The term "aryl," used alone or as part of a larger moiety as in "aralkyl," "aralkoxy," or "aryloxyalkyl," refers to monocyclic and bicyclic ring systems having a total of 5 to 14 ring members, wherein at least one ring in the system is aromatic, and wherein each ring in the system contains 3 to 7 ring members. The term "aryl" is used interchangeably with the term "aryl ring." In certain embodiments of the invention, "aryl" refers to an aromatic ring system. Exemplary aryl groups are phenyl, biphenyl, naphthyl, anthracyl, and the like, which optionally include one or more substituents. Also included within the scope of the term "aryl," as it is used herein, are groups in which an aromatic ring is fused to one or more non-aromatic rings, such as indanyl, phthalimidyl, naphthimidyl, phenanthridinyl, or tetrahydronaphthyl.

[0023] The terms "heteroaryl" and "heteroar-," used alone or as part of a larger moiety, e.g., "heteroaralkyl" or "heteroaralkoxy," refer to groups having 5 to 10 ring atoms, preferably 5, 6, or 9 ring atoms; having 6, 10, or 14 pi electrons shared in the cyclic array; and having from 1 to 5 heteroatoms in addition to carbon atoms. The term "heteroatom" refers to nitrogen, oxygen, or sulfur and includes any oxidized form of nitrogen or sulfur and the quaternized form of a basic nitrogen. Heteroaryl groups include, without limitation, thienyl, furanyl, pyrrolyl, imidazolyl, pyrazolyl, triazolyl, tetrazolyl, oxazolyl, isoxazolyl, oxadiazolyl, thiazolyl, isothiazolyl, thiadiazolyl, pyridyl, pyridazinyl, pyrimidyl, pyrazinyl, indolizinyl, purinyl, naphthyridinyl, and pteridinyl. The terms "heteroaryl" and "heteroara-," as used herein, also include groups in which a heteroaromatic ring is fused with one or more aryl, alicyclic, or heterocyclyl rings, where the radical or point of attachment is on the heteroaromatic ring. Non-limiting examples include indolyl, isoindolyl, benzothienyl, benzofuranyl, dibenzofuranyl, indazolyl, benzimidazolyl, benzthiazolyl, quinolyl, isoquinolyl, cinnolinyl, phthalazinyl, quinazolinyl, quinoxalinyl, 4H-quinolizinyl, carbazolyl, acridinyl, phenazinyl, phenothiazinyl, phenoxazinyl, tetrahydroquinolinyl, tetrahydroisoquinolinyl, and pyrido[2,3-b]-1,4-oxazin-3(4H)-one. Heteroaryl groups are optionally mono- or bicyclic. The term "heteroaryl" is used interchangeably with the terms "heteroaryl ring," "heteroaryl group," or "heteroaromatic," all of which encompass rings that are optionally substituted. The term "heteroaralkyl" refers to an alkyl group substituted by a heteroaryl, where the alkyl and heteroaryl portions independently are optionally substituted.

[0024] As used herein, the terms "heterocycle," "heterocyclyl," "heterocyclic radical," and "heterocyclic ring" are used interchangeably and refer to a stable 5- to 7-membered monocyclic or 7- to 10-membered bicyclic heterocyclic moiety, which is either saturated or partially unsaturated and, in addition to carbon atoms, has one or more, preferably one to four, heteroatoms as defined above. When used in reference to a ring atom of a heterocycle, the term "nitrogen" includes substituted nitrogen. By way of example, in a saturated or partially unsaturated ring having 0 to 3 heteroatoms selected from oxygen, sulfur, or nitrogen, the nitrogen can be N (as in 3,4-dihydro-2H-pyrrolyl), NH (as in pyrrolidinyl), or + NR (as found in N-substituted pyrrolidinyls).

[0025] A heterocycle can be attached to its pendant group at a heteroatom or a carbon atom that results in a stable structure, and any of the ring atoms can be optionally substituted. Examples of such saturated or partially unsaturated heterocyclic radicals include, without limitation, tetrahydrofuranyl, tetrahydrothiophenylpyrrolidinyl, piperidinyl, pyrrolinyl, tetrahydroquinolinyl, tetrahydroisoquinolinyl, decahydroquinolinyl, oxazolidinyl, piperazinyl, dioxanyl, dioxolanyl, diazepinyl, oxazepinyl, thiazepinyl, morpholinyl, and quinuclidinyl. The terms "heterocycle," "heterocyclyl," "heterocyclyl ring," "heterocyclic group," "heterocyclic moiety," and "heterocyclic radical" are used interchangeably herein and also include groups in which a heterocyclyl ring is fused to one or more aryl, heteroaryl, or alicyclic rings, such as indolinyl, 3H-indolyl, chromanyl, phenanthridinyl, or tetrahydroquinolinyl, where the radical or point of attachment is on the heterocyclyl ring. Heterocyclyl groups are optionally mono- or bicyclic. The term "heterocyclylalkyl" refers to an alkyl group substituted by a heterocyclyl, where the alkyl and heterocyclyl portions may independently be optionally substituted.

[0026] As used herein, the term "partially unsaturated" refers to a ring moiety that includes at least one double or triple bond. The term "partially unsaturated" is intended to encompass rings with multiple sites of unsaturation, but is not intended to include aryl or heteroaryl moieties, as defined herein.

[0027] Fused rings are described by the embodiments for each ring, Ring A and Ring B, as described herein. Together, Ring A and Ring B form a fused heteroaryl ring, as allowed by valence (e.g., Ring A is [ka] and ring B is [ka] When ring A and ring B are then together [ka] (It is).

[0028] As described herein, certain compounds of the invention include "optionally substituted" moieties. In general, the term "substituted," whether preceded by the term "optionally," means that one or more hydrogens of the specified moiety have been replaced with a suitable substituent. "Substituted" refers to the replacement of a given structure (e.g., [ka] At least [ka] refers to; and [ka] At least [ka] (referring to a group represented by a formula (I) or (II)), applies either explicitly or implicitly to one or more hydrogens. Unless so indicated, an "optionally substituted" group has a suitable substituent at each substitutable position of that group, and when more than one position in any given structure is substituted with more than one substituent selected from a specified group, the substituents may be either the same or different at every position. Combinations of substituents envisioned by the present invention are preferably those that result in the formation of stable or chemically feasible compounds. The term "stable," as used herein, refers to compounds that remain substantially unchanged when subjected to conditions that permit their production, detection, and in some embodiments, their recovery, purification, and use for one or more of the purposes disclosed herein.

[0029] Suitable monovalent substituents on a substitutable carbon atom of an "optionally substituted" group are independently deuterium; halogen; -(CH) 0~4 R°;-(CH2) 0~4 OR°;-O(CH2) 0~4 R o , -O-(CH2) 0~4 C(O)OR°;-(CH2) 0~4 CH(OR°)2;-(CH2) 0~4 SR°; optionally substituted with R°, -(CH2) 0~4 Ph; optionally substituted with R°, -(CH2) 0~4 O(CH2) 0~1 Ph; optionally substituted with R°, -CH=CHPh; optionally substituted with R°, -(CH2) 0~4 O(CH2) 0~1 -pyridyl; -NO2; -CN; -N3; ​​-(CH2) 0~4 N(R°)2;-(CH2) 0~4 N(R°)C(O)R°;-N(R°)C(S)R°;-(CH2) 0~4 N(R°)C(O)NR°2;-N(R°)C(S)NR°2;-(CH2) 0~4 N(R°)C(O)OR°;-N(R°)N(R°)C(O)R°;-N(R°)N(R°)C(O)NR°2;-N(R°)N(R°)C(O)OR°;-(CH2)0~4 C(O)R°;-C(S)R°;-(CH2) 0~4 C(O)OR°;-(CH2) 0~4 C(O)SR°;-(CH2) 0~4 C(O)OSiR°3;-(CH2) 0~4 OC(O)R°;-OC(O)(CH2) 0~4 SR°, SC(S)SR°;-(CH2) 0~4 SC(O)R°;-(CH2) 0~4 C(O)NR°2;-C(S)NR°2;-C(S)SR°;-SC(S)SR°, -(CH2) 0~4 OC(O)NR°2;-C(O)N(OR°)R°;-C(O)C(O)R°;-C(O)CH2C(O)R°;-C(NOR°)R°;-(CH2) 0~4 SSR°;-(CH2) 0~4 S(O)2R°;-(CH2) 0~4 S(O)2OR°;-(CH2) 0~4 OS(O)2R°;-S(O)2NR°2;-(CH2) 0~4 S(O)R°;-N(R°)S(O)2NR°2;-N(R°)S(O)2R°;-N(OR°)R°;-C(NH)NR°2;-P(O)2R°;-P(O)R°2;-OP(O)R°2;-OP(O)(OR°)2;SiR°3;-(C 1~4 linear or branched alkylene)ON(R°)2; or -(C 1~4 straight chain or branched alkylene)C(O)ON(R°)2, where each R° is optionally substituted as defined below, and each R° is independently hydrogen, C 1~6 Aliphatic, -CH2Ph, -O(CH2) 0~1Ph, -CH2- (a 5-6 membered heteroaryl ring), or a saturated, partially unsaturated, or aryl 5-6 membered ring (having 0-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur); or, notwithstanding the above definition, two independently occurring R° together with their intervening atom(s) form a saturated, partially unsaturated, or aryl 3-12 membered mono- or bicyclic ring (having 0-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur), which may be optionally substituted as defined below.

[0030] Suitable monovalent substituents on R° (or the ring formed by two independent occurrences of R° together with their intervening atoms) are independently deuterium, halogen, —(CH) 0~2 R l ,-(Halo R l ), -(CH2) 0~2 OH, -(CH2) 0~2 OR l , -(CH2) 0~2 CH(OR l )2;-O(HaloR l ), -CN, -N3, -(CH2 ) 0~2 C(O)R l , -(CH2) 0~2 C(O)OH, -(CH2) 0~2 C(O)OR l , -(CH2) 0~2 SR l , -(CH2) 0~2 SH, -(CH2) 0~2 NH2, -(CH2) 0~2 NHR l , -(CH2) 0~2 NR l 2, -NO2, -SiR l 3. -OSiR l 3. -C(O)SR l , -(C 1~4 Straight or branched alkylene)C(O) OR l , or -SSR l where each R lis unsubstituted or, if preceded by "halo", is substituted with only one or more halogens, and each R l independently, C 1~4 Aliphatic, -CH2Ph, -O(CH2) 0~1 or a 5-6 membered saturated, partially unsaturated, or aryl ring (having 0-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur). Suitable divalent substituents on a saturated carbon atom of R° include ═O and ═S.

[0031] Suitable divalent substituents on a saturated carbon atom of an "optionally substituted" group are the following: =O, =S, =NNR * 2, =NNHC(O)R * , =NNHC(O)OR * , =NNHS(O)2R * , =NR * , =NOR * , -O(C(R * 2)) 2~3 O-, or -S(C(R * 2)) 2~3 S-, but here R * Each independent occurrence of is selected from hydrogen, a 5-6 membered saturated, partially unsaturated, or aryl ring (having 0-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur) that is substituted or unsubstituted as defined below. Preferred divalent substituents attached to adjacent substitutable carbons of an "optionally substituted" group are: -O(CR * 2) 2~3 O-, where R * Each independent occurrence of is selected from hydrogen, a 5-6 membered saturated, partially unsaturated, or aryl ring (having 0-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur) that is substituted or unsubstituted as defined below.

[0032] R * Suitable substituents on the aliphatic groups are halogen, -R l ,-(Halo R l ), -OH, -ORl , -O(HaloR l ), -CN, -C(O)OH, -C(O)OR l , -NH2, -NHR l , -NR l 2, or -NO2, where each R l is unsubstituted, or if preceded by "halo", one or more halogens, and each R l independently, C 1~4 Aliphatic, -CH2Ph, -O(CH2) 0~1 Ph, or a 5-6 membered saturated, partially unsaturated, or aryl ring (having 0-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur).

[0033] A suitable substituent on a substitutable nitrogen of an "optionally substituted" group is -R † , -NR † 2. -C(O)R † , -C(O)OR † , -C(O)C(O)R † , -C(O)CHC(O)R † , -S(O)2R † , -S(O)NR † 2. -C(S)NR † 2. -C(NH)NR † 2, or -N(R † )S(O)2R † where each R † are independently hydrogen, optionally substituted C as defined below 1~6 an aliphatic, unsubstituted -OPh, or an unsubstituted 5-6 membered saturated, partially unsaturated, or aryl ring (having 0-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur); or, notwithstanding the above definition, R † two independent occurrences of are taken together with their intervening atom(s) to form an unsubstituted 3-12 membered saturated, partially unsaturated, or aryl mono- or bicyclic ring (having 0-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur).

[0034] R † Suitable substituents on the aliphatic group are independently halogen, —R l ,-(Halo R l ), -OH, -OR l , -O(HaloR l ), -CN, -C(O)OH, -C(O)OR l , -NH2, -NHR l , -NR l 2, or -NO2, where each R l is unsubstituted or, if preceded by "halo", is substituted with only one or more halogens, and each R l independently, C 1~4 Aliphatic, -CH2Ph, -O(CH2) 0~1 Ph, or a 5-6 membered saturated, partially unsaturated, or aryl ring (having 0-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur).

[0035] In certain embodiments, the terms "optionally substituted (optionally substituted)," "optionally substituted alkyl," "optionally substituted alkenyl," "optionally substituted alkynyl," "optionally substituted carbocyclic," "optionally substituted aryl," "optionally substituted heteroaryl," "optionally substituted heterocyclic," and any other optional substituent as used herein refer to groups that are substituted or unsubstituted by the independent replacement of one, two, or more of the hydrogen atoms on that group with exemplary substituents, including, but not limited to, the following: -F, -Cl, -Br, -I, deuterium, -OH, protected hydroxy, alkoxy, oxo, thioxo, -NO2, -CN, CF3, N3, -NH, protected amino, -NH alkyl, -NH alkenyl, -NH alkynyl, -NH cycloalkyl, -NH-aryl, -NH-heteroaryl, -NH-heterocyclic, -dialkylamino, -diarylamino, -diheteroarylamino, -O-alkyl, -O-alkenyl, -O-alkynyl, -O-cycloalkyl, -O-aryl, -O-heteroaryl, -O-heterocyclic, -C(O)-alkyl, -C(O)-alkenyl, -C(O)-alkynyl, -C(O)-carbocyclyl, -C(O)-aryl, -C(O)-heteroaryl, -C(O)-heterocyclyl, -CONH, -CONH-alkyl, -CONH-alkenyl, -CONH-alkynyl, -CONH-carbocyclyl, -CONH-aryl, -CONH-heteroaryl, -CONH-heterocyclyl, -OCO2-alkyl, -OCO2-alkenyl, -OCO2-alkynyl, -OCO2-carbocyclyl, -OCO2-aryl, -OCO2-heteroaryl, -OCO2-heterocyclyl, -OCONH2, -OCONH-alkyl, -OCONH-alkenyl, -OCONH-alkynyl, -OCONH-carbocyclyl, -OCONH-aryl, -OCONH-heteroaryl, -OCONH-heterocyclyl, -NHC(O)-alkyl, -NHC(O)-alkenyl, -NHC(O)-alkynyl, -NHC(O)-carbocyclyl, -NHC(O)-aryl, -NHC(O)-heteroaryl, -NHC(O)-heterocyclyl, -NHCO2-alkyl, -NHCO2-alkenyl, -NHCO2-alkynyl, -NHCO2-carbocyclyl, -NHCO2-aryl, -NHCO2-heteroaryl, -NHCO2-heterocyclyl, -NHC(O)NH2, -NHC(O)NH-alkyl, -NHC(O)NH-alkenyl, -NHC( O)NH-alkenyl, -NHC(O)NH-carbocyclyl, -NHC(O)NH-aryl, -NHC(O)NH-heteroaryl, -NHC(O)NH-heterocyclyl, NHC(S)NH2, -NHC(S)NH-alkyl, -NHC(S)NH-alkenyl, -NHC(S)NH-alkynyl, -NHC(S)NH-carbocyclyl, -NHC(S)NH-aryl, -NHC(S)NH-heteroaryl, -NHC(S)NH-heterocyclyl, -NHC(NH)NH2, -NHC(NH)NH-alkyl, -NHC(NH)NH- -alkenyl, -NHC(NH)NH-alkenyl, -NHC(NH)NH-carbocyclyl, -NHC(NH)NH-aryl, -NHC(NH)NH-heteroaryl, -NHC(NH)NH-heterocyclyl, -NHC(NH)-alkyl, -NHC(NH)-alkenyl, -NHC(NH)-alkenyl, -NHC(NH)-carbocyclyl, -NHC(NH)-aryl, -NHC(NH)-heteroaryl, -NHC(NH)-heterocyclyl, -C(NH)NH-alkyl, -C(NH)NH-alkenyl, -C(NH)NH-alkynyl, -C(NH)NH-carbocyclyl, -C(NH)NH-aryl, -C(NH)NH-heteroaryl, -C(NH)NH-heterocyclyl, -S(O)-alkyl, -S(O)-alkenyl, -S(O)-alkynyl, -S(O)-carbocyclyl, -S(O)-aryl, -S(O)-heteroaryl, -S(O)-heterocyclyl-SONH, -SONH-alkyl, -SONH-alkenyl, -SONH-alkynyl, -SONH-carbocyclyl, -SONH-aryl, -SONH-heteroaryl, -SONH-heterocyclyl, -NHSO2-alkyl, -NHSO2-alkenyl, -NHSO2-alkynyl, -NHSO2-carbocyclyl, -NHSO2-aryl, -NHSO2-heteroaryl, -NHSO2-heterocyclyl, -CH2NH2, -CH2SO2CH3, mono-, di-, or tri-alkylsilyl, -alkyl, -alkenyl, -alkynyl, -aryl, -arylalkyl, -heteroaryl, -heteroarylalkyl, -heterocycloalkyl, -cycloalkyl, -carbocyclic, -heterocyclic, polyalkoxyalkyl, polyalkoxy, -methoxymethoxy, -methoxyethoxy, -SH, -S-alkyl, -S-alkenyl, -S-alkynyl, -S-carbocyclyl, -S-aryl, -S-heteroaryl, -S-heterocyclyl, or methylthiomethyl.

[0036] As used herein, the term "pharmaceutically acceptable salts" refers to those salts that, within the scope of sound medical judgment, are suitable for use in contact with the tissues of humans and lower animals without undue toxicity, irritation, allergic response, etc., and are commensurate with a reasonable benefit / risk ratio. Pharmaceutically acceptable salts are well known in the art. For example, SM Berge et al. describe pharmaceutically acceptable salts in detail in J. Pharmaceutical Sciences, 1977, 66, 1-19 (incorporated herein by reference). Pharmaceutically acceptable salts of the compounds of the present invention include those derived from suitable inorganic and organic acids and bases. Examples of pharmaceutically acceptable non-toxic acid addition salts are salts of amino groups formed with inorganic acids such as hydrochloric, hydrobromic, phosphoric, sulfuric, and perchloric acids, or with organic acids such as acetic, oxalic, maleic, tartaric, citric, succinic, or malonic acids, or by using other methods used in the art, such as ion exchange. Other pharmaceutically acceptable salts include those with adipic, alginic, ascorbic, aspartic, benzenesulfonic, benzoic, bisulfuric, boric, butyric, camphoric, camphorsulfonic, citric, cyclopentanepropionic, digluconic, dodecylsulfuric, ethanesulfonic, formic, fumaric, glucoheptonic, glycerophosphate, gluconic, hemisulfuric, heptanoic, hexanoic, hydroiodic, 2-hydroxy-ethanesulfonic, and the like. Included are salts of sulfonic acid, lactobionic acid, lactic acid, lauric acid, lauryl sulfate, malic acid, maleic acid, malonic acid, methanesulfonic acid, 2-naphthalenesulfonic acid, nicotinic acid, nitric acid, oleic acid, oxalic acid, palmitic acid, pamoic acid, pectinic acid, persulfuric acid, 3-phenylpropionic acid, phosphoric acid, pivalic acid, propionic acid, stearic acid, succinic acid, sulfuric acid, tartaric acid, thiocyanic acid, p-toluenesulfonic acid, undecanoic acid, and valeric acid.

[0037] Salts derived from appropriate bases include alkali metal, alkaline earth metal, ammonium and N + (C1~4 Representative alkali metal or alkaline earth metal salts include sodium, lithium, potassium, calcium, magnesium, and the like. Further pharmaceutically acceptable salts include non-toxic ammonium, quaternary ammonium, and amine cations formed using counterions such as halides, hydroxides, carboxylates, sulfates, phosphates, nitrates, lower alkyl and aryl sulfonates, when appropriate.

[0038] Unless so stated, structures depicted herein are also meant to encompass all isomeric (e.g., enantiomeric, diastereomeric, and geometric (or conformational)) forms of that structure; for example, the R and S configurations, Z and E double bond isomers, and Z and E conformational isomers for each asymmetric center. Thus, single stereochemical isomers as well as enantiomeric, diastereomeric, and geometric (or conformational) mixtures of the present compounds are within the scope of the invention. Unless so stated, all tautomeric forms of the compounds of the invention are within the scope of the invention.

[0039] Additionally, unless so stated, structures depicted herein are also meant to include compounds that differ only in the presence of one or more isotopically enriched atoms, for example, the replacement of hydrogen by deuterium or tritium, or 13 C is or 14 Compounds having this structure including the replacement of a carbon with a C-enriched carbon are within the scope of this invention. In some embodiments, the group comprises one or more deuterium atoms.

[0040] It is further intended that the compounds of Formula I encompass isotopically labeled forms thereof. Isotopically labeled forms of compounds of Formula I are identical to the compounds except for the fact that one or more atoms of the compound have been replaced by an atom or atoms having an atomic mass or mass number different from the atomic mass or mass number of the normally occurring atom. Examples of isotopes that are readily commercially available and that can be incorporated into compounds of Formula I by well-known methods include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorus, fluorine, and chlorine, for example, 2 H, 3 H, 13 C. 14 C. 15 N, 18 O. 17 O. 31 P, 32 P, 35 S, 18 F and 36 CI. Compounds of Formula I, their prodrugs, or any pharmaceutically acceptable salts thereof containing one or more of the above isotopes and / or other isotopes of other atoms are intended to be part of the present invention. Isotopically labeled compounds of Formula I can be used in a number of beneficial ways. For example, isotopically labeled compounds of Formula I can be used in, for example, 3 H or 14 Radioactive isotopes such as C have been incorporated therein and are suitable for drug and / or substrate tissue distribution assays. 3 H) and carbon-14 ( 14 C) is particularly preferred due to its easy preparation and excellent detectability. Heavier isotopes, such as deuterium ( 2Incorporation of H) into compounds of Formula I has therapeutic advantages due to the higher metabolic stability of the isotopically labeled compounds. Higher metabolic stability translates directly into increased in vivo half-life or smaller dosages, which under most circumstances represent preferred aspects of the present invention. Isotopically labeled compounds of Formula I can generally be prepared by following the procedures disclosed in the synthetic schemes and related descriptions in the Examples and Preparations sections herein, substituting readily available isotopically labeled reactants for non-isotopically labeled reactants.

[0041] deuterium( 2 H) can also be incorporated into compounds of Formula I for the purpose of manipulating the oxidative metabolism of the compound through the primary kinetic isotope effect. The primary kinetic isotope effect is the change in the rate of a chemical reaction due to the exchange of an isotopic nuclei, which in turn is caused by the change in ground state energy required for covalent bond formation after this isotope exchange. The exchange of a heavier isotope usually results in a lowering of the ground state energy of the chemical bond, thus causing a slowing of the rate-limiting bond cleavage. When bond cleavage occurs along the coordinate of a multi-product reaction in or near a saddle-point region, the product distribution ratio can be substantially altered. By way of illustration: when deuterium is attached to a carbon atom at a non-exchangeable position, k M / k D A rate difference of 2 to 7 is typical. If this rate difference is successfully applied to an oxidation-susceptible compound of formula I, the in vivo profile of the compound can be dramatically modified, resulting in improved pharmacokinetic properties.

[0042] When discovering and developing therapeutic agents, it is reasonable for one skilled in the art to assume that many compounds with poor pharmacokinetic profiles are susceptible to oxidative metabolism, allowing for optimization of pharmacokinetic parameters while retaining desirable in vitro properties. Currently available in vitro liver microsome assays provide valuable information about the course of this type of oxidative metabolism, which in turn allows for the rational design of deuterated compounds of Formula I with improved stability through resistance to such oxidative metabolism. Significant improvements in the pharmacokinetic profile of compounds of Formula I can thereby be obtained, including improvements in in vivo half-life (t / 2), concentration at maximum therapeutic effect (C max ), area under the dose-response curve (AUC), and F; and quantitatively in terms of reduced clearance, dose, and material cost.

[0043] The following is intended to illustrate the above: A compound of Formula I having multiple potential sites of oxidative metabolic attack (e.g., benzylic hydrogen atoms and hydrogen atoms attached to nitrogen atoms) is prepared as a series of analogs in which various combinations of hydrogen atoms are replaced by deuterium atoms (such that some, most, or all of these hydrogen atoms are replaced by deuterium atoms). Determination of half-life allows for a convenient and accurate determination of the extent to which resistance to oxidative metabolism has improved. In this way, it is determined that the half-life of the parent compound can be extended by up to 100% as a result of this type of deuterium-hydrogen exchange.

[0044] Deuterium-hydrogen exchange in compounds of Formula I can also be used to achieve favorable modification of the metabolic spectrum of the starting compound to reduce or eliminate undesired toxic metabolites. For example, if a toxic metabolite arises through oxidative carbon-hydrogen (CH) bond cleavage, it can be reasonably expected that a deuterated analog will greatly reduce or eliminate the production of the unwanted metabolite, even if the specific oxidation is not the rate-limiting step. Further information on the state of the art regarding deuterium-hydrogen exchange can be found, for example, in Hanzlik et al., J. Org. Chem. 55, 3992-3997, 1990; Reider et al., J. Org. Chem. 52, 3326-3334, 1987; Foster, Adv. Drug Res. 14, 1-40, 1985; Gillette et al., Biochemistry 33(10) 2927-2937, 1994; and Jarman et al., Carcinogenesis 16(4), 683-688, 1993.

[0045] As used herein, the term "modulator" is defined as a compound that binds to and / or inhibits a target with measurable affinity. In certain embodiments, a modulator has an IC of less than about 50 μM, less than about 1 μM, less than about 500 nM, less than about 100 nM, or less than about 10 nM. 50 and / or have a binding constant.

[0046] The terms "measurable affinity" and "measurably inhibit," as used herein, refer to a measurable change in TLR7 / 8 activity between a sample containing a compound of the present invention or a composition thereof and TLR7 / 8 (and an equivalent sample containing TLR7 / 8 in the absence of the compound or composition thereof).

[0047] Combinations of substituents and variants envisioned by this invention are only those that result in the formation of stable compounds. The term "stable," as used herein, refers to a compound that possesses sufficient stability to permit its manufacture and maintains its integrity for a sufficient period of time to be useful for the purposes detailed herein (e.g., therapeutic or prophylactic administration to a subject).

[0048] The recitation of a list of chemical groups in any definition of a variant herein includes definitions of that variant as any single group or combination of the groups in the list. The recitation of an embodiment for a variant herein includes that embodiment as any single embodiment or in combination with any other embodiment or portion thereof.

[0049] 3. Description of example compounds According to one aspect, the present invention provides a compound of formula I: [ka] or a pharmaceutically acceptable salt thereof, wherein: Ring A is aryl or heteroaryl having 1 to 4 heteroatoms (independently selected from nitrogen, oxygen, and sulfur), each of which may be optionally substituted; Ring B is a heteroaryl having 1 to 4 heteroatoms (independently selected from nitrogen, oxygen, and sulfur), each of which may be optionally substituted; R 1 is -CH3, -CF3, or -CN; Each R 2 are independently -H, -R, halogen, -haloalkyl, -OR, -SR, -CN, -NO2, -S2R, -SOR, -C(O)R, -C2R, -C(O)N(R)2, -NRC(O)R, -NRC(O)N(R)2, -NRSO2R, or -N(R)2; Each R 3are independently -H, -R, halogen, -haloalkyl, -OR, -SR, -CN, -NO2, -S2R, -SOR, -C(O)R, -C2R, -C(O)N(R)2, -NRC(O)R, -NRC(O)N(R)2, -NRSO2R, or -N(R)2; Each R 4 are independently -H, -R, halogen, -haloalkyl, -OR, -SR, -CN, -NO2, -S2R, -SOR, -C(O)R, -C2R, -C(O)N(R)2, -NRC(O)R, -NRC(O)N(R)2, -NRC(O)OR, -NRSO2R, or -N(R)2; Each R 5 are independently -H, -R, halogen, -haloalkyl, -OR, -SR, -CN, -NO2, -S2R, -SOR, -C(O)R, -C2R, -C(O)N(R)2, -NRC(O)R, -NRC(O)N(R)2, -NRSO2R, or -N(R)2; Each R is independently hydrogen, C 1~6 aliphatic, C 3~10 aryl, a 3-8 membered saturated or partially unsaturated carbocycle, a 3-7 membered heterocycle having 1-4 heteroatoms (independently selected from nitrogen, oxygen, or sulfur), or a 5-6 membered monocyclic heteroaryl ring having 1-4 heteroatoms (independently selected from nitrogen, oxygen, or sulfur); each of which may be optionally substituted; or Two R groups on the same atom, together with the atom to which they are attached, form a C 3~10 forming an aryl, a 3-8 membered saturated or partially unsaturated carbocycle, a 3-7 membered heterocycle having 1-4 heteroatoms (independently selected from nitrogen, oxygen, or sulfur), or a 5-6 membered monocyclic heteroaryl ring having 1-4 heteroatoms (independently selected from nitrogen, oxygen, or sulfur); each of which may be optionally substituted; k is 0, 1, or 2; n is 0, 1, or 2; p is 0, 1, or 2; r is 1, 2, or 3; and t is 1, 2, or 3. where k is 0 when linked to nitrogen; and k is 1 or 2 when linked to carbon.

[0050] In some embodiments, R 1 is -CH3.

[0051] In some embodiments, R 1 is -CF3.

[0052] In some embodiments, R 1 is -CN.

[0053] In some embodiments, k is 0, and R 1 does not exist.

[0054] In some embodiments, ring A is phenyl or a 6-membered monocyclic heteroaryl having 1 to 4 heteroatoms (independently selected from nitrogen, oxygen, or sulfur).

[0055] In some embodiments, ring A is phenyl, pyridyl, pyrimidyl, pyrazinyl, pyridazinyl, or triazinyl.

[0056] In some embodiments, ring A is phenyl or pyridyl.

[0057] In some embodiments, ring A is [ka] is.

[0058] In some embodiments, ring A is [ka] is.

[0059] In some embodiments, ring A is [ka] is.

[0060] In some embodiments, ring A is [ka] is.

[0061] In some embodiments, ring A is [ka] is.

[0062] In some embodiments, ring A is [ka] is.

[0063] In some embodiments, ring A is [ka] is.

[0064] In some embodiments, Ring B is a 5-6 membered monocyclic heteroaryl having 1-4 heteroatoms (independently selected from nitrogen, oxygen, and sulfur).

[0065] In some embodiments, Ring B is pyridyl, pyrimidyl, pyrazinyl, pyridazinyl, triazinyl, pyrrole, imidazole, isoxazole, oxazole, or thiazole; each of which may be optionally substituted.

[0066] In some embodiments, Ring B is pyridyl, pyrazinyl, or pyrrole; each of which may be optionally substituted.

[0067] In some embodiments, ring B is [ka] is.

[0068] In some embodiments, ring B is [ka] is.

[0069] In some embodiments, ring B is [ka] is.

[0070] In some embodiments, ring B is [ka] is.

[0071] In some embodiments, ring B is [ka] is.

[0072] In some embodiments, each R 2 are independently -H.

[0073] In some embodiments, each R 2 independently C 1-6 aliphatic, C 3-10 aryl, a 3-8 membered saturated or partially unsaturated carbocyclic ring, a 3-7 membered heterocyclic ring having 1-4 heteroatoms (independently selected from nitrogen, oxygen, or sulfur), or a 5-6 membered monocyclic heteroaryl ring having 1-4 heteroatoms (independently selected from nitrogen, oxygen, or sulfur); each of which may be optionally substituted.

[0074] In some embodiments, each R 2 are independently methyl, ethyl, ethyl, propyl, i-propyl, butyl, s-butyl, t-butyl, straight-chained or branched pentyl, or straight-chained or branched hexyl; each of which may be optionally substituted.

[0075] In some embodiments, each R 2are independently selected from phenyl, naphthyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, adamantyl, cyclooctyl, [3.3.0]bicyclooctanyl, [4.3.0]bicyclononanyl, [4.4.0]bicyclodecanyl, [2.2.2]bicyclooctanyl, fluorenyl, indanyl, tetrahydronaphthyl, acridinyl, azocinyl, benzimidazolyl, benzofuranyl, benzothiofuranyl, benzothiophenyl, benzoxazolyl, benzthiazolyl, benztriazolyl, benztetrazolyl, benzisoxazolyl, benzisothiazolyl, benzimidazolinyl, carbazolyl, NH-carbazolyl, carbolinyl inyl), chromanyl, chromenyl, cinnolinyl, decahydroquinolinyl, 2H,6H-1,5,2-dithiazinyl, dihydrofuro[2,3-b]tetrahydrofuran, furanyl, furazanyl, imidazolidinyl, imidazolinyl, imidazolyl, 1H-indazolyl, indolenyl, indolinyl, indolizinyl, indolyl, 3H-indolyl, isoindolinyl, isoindolenyl, isobenzofuranyl, isochromanyl, isoindazolyl, isoindolinyl, isoindolyl, isoquinolinyl, isothiazolyl, isoxazolyl, morpholinyl, naphthyridinyl, octahydroisoquinolinyl, oxadiazolyl, 1,2,3-oxadiazolyl, 1,2,4-oxadiazolyl;-1,2,5-oxadiazolyl, 1,3,4-oxadiazolyl, oxazolidinyl, oxazolyl, oxazolidinyl, pyrimidinyl, phenanthridinyl, phenanthrolinyl, phenazinyl, phenothiazinyl, phenoxathiinyl, phenoxazinyl, phthalazinyl, piperazinyl, piperidinyl, pteridinyl, purinyl, pyranyl, pyrazinyl, pyrazolidinyl, pyrazolinyl, pyrazolyl, pyridazinyl, pyridooxazole, pyridoimidazole, pyridothiazole, pyridinyl, pyridyl, pyrimidinyl, pyrrolidinyl, pyrrolinyl, 2H-pyrrolyl, pyrrolyl, quinazolinyl, quinolinyl, 4H-quinolizinyl, quinolyl, oxalinyl, quinuclidinyl, tetrahydrofuranyl, tetrahydroisoquinolinyl, tetrahydroquinolinyl, 6H-1,2,5-thiadiazinyl, 1,2,3-thiadiazolyl, 1,2,4-thiadiazolyl, 1,2,5-thiadiazolyl, 1,3,4-thiadiazolyl, thianthrenyl, thiazolyl, thienyl, thienothiazolyl, thienoxazolyl, thienoimidazolyl, thiophenyl, triazinyl, 1,2,3-triazolyl, 1,2,4-triazolyl, 1,2,5-triazolyl, 1,3,4-triazolyl, oxetanyl, azetidinyl, or xanthenyl; each of which may be optionally substituted;

[0076] In some embodiments, each R 2 are independently halogen, -haloalkyl, -OR, -SR, -CN, -NO, -SOR, -SOR, -C(O)R, -COR, -C(O)N(R), -NRC(O)R, -NRC(O)N(R), -NRSOR, or -N(R).

[0077] In some embodiments, each R 3 are independently -H.

[0078] In some embodiments, each R 3 independently C 1-6 aliphatic, C 3-10aryl, a 3-8 membered saturated or partially unsaturated carbocyclic ring, a 3-7 membered heterocyclic ring having 1-4 heteroatoms (independently selected from nitrogen, oxygen, or sulfur), or a 5-6 membered monocyclic heteroaryl ring having 1-4 heteroatoms (independently selected from nitrogen, oxygen, or sulfur); each of which may be optionally substituted.

[0079] In some embodiments, each R 3 are independently methyl, ethyl, ethyl, propyl, i-propyl, butyl, s-butyl, t-butyl, straight-chained or branched pentyl, or straight-chained or branched hexyl; each of which may be optionally substituted.

[0080] In some embodiments, each R 3are independently selected from the group consisting of phenyl, naphthyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, adamantyl, cyclooctyl, [3.3.0]bicyclooctanyl, [4.3.0]bicyclononanyl, [4.4.0]bicyclodecanyl, [2.2.2]bicyclooctanyl, fluorenyl, indanyl, tetrahydronaphthyl, acridinyl, azocinyl, benzimidazolyl, benzofuranyl, benzothiofuranyl, benzothiophenyl, benzoxazolyl, benzthiazolyl, benztriazolyl, benztetrazolyl, benzisoxazolyl, benzisothiazolyl, benzimidazolinyl, carbazolyl, NH-carbazolyl, carbolinyl, chloazolyl, chlorin ... rhomanyl, chromenyl, cinnolinyl, decahydroquinolinyl, 2H,6H-1,5,2-dithiazinyl, dihydrofuro[2,3-b]tetrahydrofuran, furanyl, furazanyl, imidazolidinyl, imidazolinyl, imidazolyl, 1H-indazolyl, isoindolinyl, indolinyl, indolizinyl, indolyl, 3H-indolyl, isoindolinyl, isoindolenyl, isobenzofuranyl, isochromanyl, isoindazolyl, isoindolinyl, isoindolyl, isoquinolinyl, isothiazolyl, isoxazolyl, morpholinyl, naphthyridinyl, octahydroisoquinolinyl, oxadiazolyl, 1,2,3-oxadiazolyl, 1,2,4-oxadiazolyl;-1,2,5-oxadiazolyl, 1,3,4-oxadiazolyl, oxazolidinyl, oxazolyl, oxazolidinyl, pyrimidinyl, phenanthridinyl, phenanthrolinyl, phenazinyl, phenothiazinyl, phenoxathiinyl, phenoxazinyl, phthalazinyl, piperazinyl, piperidinyl, pteridinyl, purinyl, pyranyl, pyrazinyl, pyrazolidinyl, pyrazolinyl, pyrazolyl, pyridazinyl, pyridooxazole, pyridoimidazole, pyridothiazole, pyridinyl, pyridyl, pyrimidinyl, pyrrolidinyl, pyrrolinyl, 2H-pyrrolyl, pyrrolyl, quinazolinyl, quinolinyl, 4H-quinolizinyl, quinolyl, oxalinyl, quinuclidinyl, tetrahydrofuranyl, tetrahydroisoquinolinyl, tetrahydroquinolinyl, 6H-1,2,5-thiadiazinyl, 1,2,3-thiadiazolyl, 1,2,4-thiadiazolyl, 1,2,5-thiadiazolyl, 1,3,4-thiadiazolyl, thianthrenyl, thiazolyl, thienyl, thienothiazolyl, thienoxazolyl, thienoimidazolyl, thiophenyl, triazinyl, 1,2,3-triazolyl, 1,2,4-triazolyl, 1,2,5-triazolyl, 1,3,4-triazolyl, oxetanyl, azetidinyl, or xanthenyl; each of which may be optionally substituted;

[0081] In some embodiments, each R 3 are independently halogen, -haloalkyl, -OR, -SR, -CN, -NO, -SOR, -SOR, -C(O)R, -COR, -C(O)N(R), -NRC(O)R, -NRC(O)N(R), -NRSOR, or -N(R).

[0082] In some embodiments, each R 4 are independently -H.

[0083] In some embodiments, each R 4 independently C 1-6aliphatic, halogen, -haloalkyl, -OR, -SR, -CN, -NO2, -S2R, -SOR, -C(O)R, -C2R, -C(O)N(R)2, -NRC(O)R, -NRC(O)N(R)2, -NRC(O)OR, -NRC(O)OR, -NRSO2R, or -N(R)2.

[0084] In some embodiments, each R 4 independently C 1-6 Aliphatic, -OR, -C(O)R, -COR, -C(O)N(R), -NRC(O)R, -NRC(O)OR, -NRC(O)N(R), -NRSOR, or -N(R); each of which may be optionally substituted.

[0085] In some embodiments, each R 4 independently C 1-6 aliphatic, -C(O)N(R)2, -NR C(O)R, or -N(R)2; each of which may be optionally substituted.

[0086] In some embodiments, each R 4 independently [ka] [ka] is.

[0087] In some embodiments, each R 5 are independently -H.

[0088] In some embodiments, each R 5 are independently halogens, C 1-6 aliphatic, C 3-10aryl, a 3-8 membered saturated or partially unsaturated carbocyclic ring, a 3-7 membered heterocyclic ring having 1-4 heteroatoms (independently selected from nitrogen, oxygen, or sulfur), or a 5-6 membered monocyclic heteroaryl ring having 1-4 heteroatoms (independently selected from nitrogen, oxygen, or sulfur); each of which may be optionally substituted.

[0089] In some embodiments, each R 5 are independently methyl, ethyl, propyl, i-propyl, butyl, s-butyl, t-butyl, straight-chained or branched pentyl, or straight-chained or branched hexyl; each of which may be optionally substituted; or each R 5 are independently -F, -Cl, -Br, or -I.

[0090] In some embodiments, each R 5 are independently methyl, ethyl, -F, -Cl, or -Br.

[0091] In some embodiments, each R 5 independently [ka] is.

[0092] In some embodiments, ring A, ring B, R 1 , R 2 , R 3 , R 4 , R 5 , k, n, p, r, and t are each as defined above and as described alone or in combination in the embodiments, classes and subclasses above and herein.

[0093] In one embodiment, the present invention provides a compound of formula Ia [ka] or a pharmaceutically acceptable salt thereof, wherein R, R 1 , R4 , R 5 , r, and t are each as defined above and as described alone or in combination in the embodiments, classes and subclasses above and herein.

[0094] In one embodiment, the present invention provides a compound of formula Ib [ka] or a pharmaceutically acceptable salt thereof, wherein R 2 , R 3 , R 4 , R 5 , n, p, r, and t are each as defined above and as described alone or in combination in the embodiments, classes and subclasses above and herein.

[0095] In one embodiment, the present invention provides a compound of formula Ic [ka] or a pharmaceutically acceptable salt thereof, wherein R 2 , R 3 , R 4 , R 5 , n, p, r, and t are each as defined above and as described alone or in combination in the embodiments, classes and subclasses above and herein.

[0096] In one embodiment, the present invention provides: Formula Ic [ka] or a pharmaceutically acceptable salt thereof, wherein R 1 , R 2 , R 3 , R 4 , R 5, k, n, p, r, and t are each as defined above and as described alone or in combination in the embodiments, classes and subclasses above and herein.

[0097] In one embodiment, the present invention provides a compound of formula Id [ka] or a pharmaceutically acceptable salt thereof, wherein R 1 , R 2 , R 3 , R 4 , R 5 , k, n, p, r, and t are each as defined above and as described alone or in combination in the embodiments, classes and subclasses above and herein.

[0098] In one embodiment, the present invention provides a compound of formula Ie: [ka] or a pharmaceutically acceptable salt thereof, wherein R 1 , R 2 , R 3 , R 4 , R 5 , k, n, p, r, and t are each as defined above and as described alone or in combination in the embodiments, classes and subclasses above and herein.

[0099] In certain embodiments, the present invention provides a compound selected from Table 1. Table 1 [Table 1] [Table 2] [Table 3] [Table 4] [Table 5] [Table 6] [Table 7] [Table 8] [Table 9] [Table 10] [Table 11] [Table 12] [Table 13] [Table 14]

[0100] In some embodiments, the present invention provides a compound selected from those depicted above, or a pharmaceutically acceptable salt thereof.

[0101] Various structural depictions may show heteroatoms without attached groups, radicals, charges, or counterions. Those skilled in the art will recognize that such depictions are intended to indicate that the heteroatom is attached to a hydrogen. (For example, [ka] teeth, [ka] is understood to be).

[0102] In certain embodiments, compounds of the present invention were synthesized according to the schemes provided in the examples below.

[0103] 4. Uses, Formulation and Administration Pharmaceutically acceptable compositions According to another aspect, the present invention provides a composition comprising a compound of the present invention or a pharmaceutically acceptable derivative thereof and a pharmaceutically acceptable carrier, adjuvant, or vehicle. The amount of the compound in the composition of the present invention is an amount effective to measurably inhibit TLR7 / 8 or a mutant thereof in a biological sample or in a patient. In some embodiments, the amount of the compound in the composition of the present invention is an amount effective to measurably inhibit TLR7 / 8 or a mutant thereof in a biological sample or in a patient. In some embodiments, the composition of the present invention is formulated for administration to a patient in need of such a composition.

[0104] The terms "patient" or "subject", as used herein, mean an animal, preferably a mammal, and most preferably a human.

[0105] The term "pharmaceutically acceptable carrier, adjuvant, or vehicle" refers to a non-toxic carrier, adjuvant, or vehicle that does not impair the pharmacological activity of the compound with which it is formulated. Pharmaceutically acceptable carriers, adjuvants, or vehicles used in the compositions of the present invention include, but are not limited to, ion exchangers, alumina, aluminum stearate, lecithin, serum proteins such as human serum albumin, buffer substances such as phosphates, glycine, sorbic acid, potassium sorbate, partial glyceride mixtures of saturated vegetable fatty acids, water, salts or electrolytes such as protamine sulfate, disodium hydrogen phosphate, potassium hydrogen phosphate, sodium chloride, zinc salts, colloidal silica, magnesium trisilicate, polyvinylpyrrolidone, cellulose-based substances, polyethylene glycol, sodium carboxymethylcellulose, polyacrylates, waxes, polyethylene-polyoxypropylene-block polymers, polyethylene glycol, and wool fat.

[0106] "Pharmaceutically acceptable derivative" means any non-toxic salt, ester, salt of an ester, or other derivative of a compound of the present invention that, upon administration to a recipient, is capable of providing, either directly or indirectly, the compound of the present invention or an inhibitory active metabolite or residue thereof.

[0107] The compositions of the present invention may be administered orally, parenterally, by inhalation spray, topically, rectally, nasally, buccally, vaginally, or via an implanted reservoir. The term "parenteral," as used herein, includes subcutaneous, intravenous, intramuscular, intra-articular, intra-synovial, intracisternal, intrathecal, intrahepatic, intralesional, and intracranial injection or infusion techniques. Preferably, the compositions are administered orally, intraperitoneally, or intravenously. Sterile injectable forms of the compositions of the present invention include aqueous or oleaginous suspensions. These suspensions are formulated according to techniques known in the art using suitable dispersing or wetting agents and suspending agents. Sterile injectable preparations may also be sterile injectable solutions or suspensions in non-toxic parenterally acceptable diluents or solvents, such as solutions in 1,3-butanediol. Among the acceptable vehicles and solvents that may be employed are water, Ringer's solution, and isotonic sodium chloride solution.In addition, sterile, fixed oils are conventionally employed as a solvent or suspending medium.

[0108] For this purpose, any bland, fixed oil may be employed, including synthetic mono- or diglycerides. Fatty acids, such as oleic acid and its glyceride derivatives, are useful in the preparation of injectables, including natural pharmaceutically acceptable oils such as olive oil or castor oil, especially their polyoxyethylated versions. Solutions or suspensions in these oils may also contain long-chain alcohol diluents or dispersants, such as carboxymethylcellulose or similar dispersants, commonly used in the formulation of pharmaceutically acceptable dosage forms, including emulsions and suspensions. Other commonly used surfactants, such as Tween, Span, and other emulsifiers or bioavailability enhancers, commonly used in the production of pharmaceutically acceptable solid, liquid, or other dosage forms, may also be used for formulation purposes.

[0109] The pharmaceutically acceptable compositions of this invention are orally administered in any orally acceptable dosage form. Exemplary oral dosage forms are capsules, tablets, aqueous suspensions or solutions. In the case of tablets for oral use, commonly used carriers include lactose and corn starch.Lubricants such as magnesium stearate are also typically added.Useful diluents for oral administration in capsule form include lactose and dried corn starch.When aqueous suspension is required for oral use, active ingredient is combined with emulsifier and suspending agent.If desired, some sweetener, flavoring agent or coloring agent can also be optionally added.

[0110] Alternatively, the pharmaceutically acceptable compositions of the present invention may be administered in the form of suppositories for rectal administration. These may be prepared by mixing the agent with a suitable non-irritating excipient that is solid at room temperature but liquid at rectal temperature, and will melt in the rectum to release the drug. Such materials include cocoa butter, beeswax, and polyethylene glycol.

[0111] The pharmaceutically acceptable compositions of this invention may also be administered topically, particularly when the target of treatment includes areas or organs readily accessible by topical application, including diseases of the eye, the skin, or the lower intestinal tract. Suitable topical formulations are readily prepared for each of these areas or organs.

[0112] Topical application for the lower intestinal tract can be effected in a rectal suppository formulation (see above) or in a suitable enema formulation. Topically-transdermal patches may also be used.

[0113] The pharmaceutically acceptable composition provided for topical application can be formulated into a suitable ointment, which contains the active ingredient suspended or dissolved in one or more carriers.The exemplary carriers for topical administration of this compound are mineral oil, liquid petrolatum, white petrolatum, propylene glycol, polyoxyethylene, polyoxypropylene compound, emulsifying wax and water.Alternatively, the pharmaceutically acceptable composition provided can be formulated into a suitable lotion or cream, which contains the active ingredient suspended or dissolved in one or more pharmaceutically acceptable carriers.Suitable carriers include but are not limited to mineral oil, sorbitan monostearate, polysorbate 60, cetyl ester wax, cetearyl alcohol, 2-octyldodecanol, benzyl alcohol and water.

[0114] The pharmaceutically acceptable compositions of this invention are optionally administered by nasal aerosol or inhalation. Such compositions are prepared according to techniques well known in the art of pharmaceutical formulation, and are prepared as solutions in saline, employing benzyl alcohol or other suitable preservatives, absorption promoters to enhance bioavailability, fluorinated carbons, and / or other conventional solubilizing or dispersing agents.

[0115] The pharmaceutically acceptable composition of the present invention is formulated for oral administration. Such formulations may be administered with or without food. In some embodiments, the pharmaceutically acceptable composition of the present invention is administered without food. In other embodiments, the pharmaceutically acceptable composition of the present invention is administered with food.

[0116] The amount of the compounds of the present invention that are optionally combined with a carrier material to produce a composition in a single dosage form will vary depending upon the host treated, the particular mode of administration, etc. Preferably, the provided compositions should be formulated so that a dosage of between 0.01-100 mg / kg body weight / day of the compound can be administered to a patient receiving these compositions.

[0117] It should also be understood that the specific dosage and treatment regimen for any particular patient will depend on a variety of factors, including the activity of the particular compound employed, the patient's age, body weight, general health, sex, diet, time of administration, rate of excretion, drug combination, and the judgment of the treating physician, and the severity of the particular disease being treated. The amount of a compound of the invention in a composition will also depend on the particular compound in the composition.

[0118] Uses of the Compounds and Pharmaceutically Acceptable Compositions The present invention also relates to a method for treating a subject suffering from a TLR7 / 8-associated disorder, said method comprising administering to said subject an effective amount of a compound of Formula I and related formulas.

[0119] The compounds of the present invention are useful as anti-cancer agents for cancers that respond to TLR7 activation.In some embodiments, cancers include but are not limited to cancers of breast, bladder, bone, brain, central and peripheral nervous system, colon, endocrine glands, esophagus, endometrium, germ cell, head and neck, kidney, liver, lung, larynx and hypopharynx, mesothelioma, sarcoma, ovary, pancreas, prostate, rectum, renal, small intestine, soft tissue, testis, stomach, skin, ureter, vagina and vulva; hereditary cancer, retinoblastoma and Wilms' tumor; leukemia, lymphoma, non-Hodgkin's disease, chronic and acute myeloid leukemia, acute lymphoblastic leukemia, Hodgkin's disease, multiple myeloma and T-cell lymphoma; myelodysplastic syndrome, plasma cell neoplasm, paraneoplastic syndrome, cancer of unknown primary site and AIDS-related malignancies.

[0120] In some embodiments, the compounds of the present invention are used to treat cancers of the skin or kidney. The sensitivity of a given cancer to TLR7 activation can be assessed by, but is not limited to, measuring a reduction (minimal, partial, or complete) in primary or metastatic tumor load, an altered blood picture, altered hormone or cytokine blood levels, inhibition of further tumor load growth, stabilization of disease in a patient, assessment of a disease-associated biomarker or surrogate marker, prolonged overall survival in a patient, prolonged time to disease progression in a patient, prolonged progression-free survival in a patient, prolonged disease-free survival in a patient, improved quality of life in a patient, or modulation of disease comorbidities (such as, but not limited to, pain, cachexia, mobilization, hospitalization, altered blood picture, weight loss, wound healing, fever).

[0121] Compounds according to the present invention may also be useful as immune response modifiers, which may modulate the immune response from a number of different angles, making them useful in the treatment of a variety of disorders.

[0122] Provided herein are methods for inhibiting an immune response in an individual, comprising administering to the individual an effective amount of an inhibitor of TLR7 and / or TLR8 (e.g., a TLR inhibitor) using a compound as described herein. In some variations, the TLR inhibitor inhibits a TLR7-dependent immune response. In some variations, the TLR inhibitor inhibits a TLR8-dependent immune response. In some variations, the TLR inhibitor inhibits a TLR7-dependent and a TLR8-dependent immune response. In some variations, the TLR inhibitor inhibits a TLR7-dependent, a TLR8-dependent, and another TLR-dependent immune response. Unless so noted, the term TLR inhibitor refers to any one of the TLR inhibitors disclosed herein. In some preferred embodiments, the individual is a human patient.

[0123] Methods of immunomodulation are provided by the present disclosure and include those that suppress and / or inhibit immune responses, including, but not limited to, immune responses. The present disclosure also provides methods for ameliorating symptoms associated with unwanted immune activation, including, but not limited to, symptoms associated with autoimmunity. Immune suppression and / or inhibition according to the methods described herein may be practiced on individuals, including those suffering from disorders associated with unwanted activation of the immune response. The present disclosure also provides methods for inhibiting TLR7- and / or TLR8-induced responses (e.g., in vitro or in vivo). In some variations, cells are contacted with a TLR inhibitor in an amount effective to inhibit a response from cells that contribute to the immune response.

[0124] Inhibition of TLR7 and / or TLR8 is useful for treating and / or preventing various diseases or disorders that respond to cytokines. Conditions for which TLR7 and / or TLR8 inhibitors may be used as treatments include, but are not limited to, autoimmune diseases and inflammatory disorders. Provided herein is a method for treating or preventing a disease or disorder in an individual, comprising administering to the individual an effective amount of a TLR7 and / or TLR8 inhibitor. Also provided is a method for ameliorating symptoms associated with a disease or disorder, comprising administering to an individual having the disease or disorder an effective amount of a TLR7 and / or TLR8 inhibitor. Also provided herein is a method for preventing or delaying the onset of a disease or disorder, comprising administering to an individual having the disease or disorder an effective amount of one or more inhibitors of TLR7 and / or TLR8. In some embodiments, the inhibitor is a compound as described herein.

[0125] Provided herein are methods of inhibiting an immune response in an individual, the methods comprising administering to the individual at least one TLR inhibitor as disclosed herein in an amount effective to inhibit the immune response in the individual. In some variations, the immune response is associated with an autoimmune disease. In a further aspect, wherein inhibiting the immune response ameliorates one or more symptoms of the autoimmune disease. In yet a further aspect, wherein inhibiting the immune response treats the autoimmune disease. In an even further aspect, wherein inhibiting the immune response prevents or delays the onset of the autoimmune disease. In some variations, the TLR inhibitor inhibits a TLR7-dependent immune response. In some variations, the TLR inhibitor inhibits a TLR8-dependent immune response. In some variations, the TLR inhibitor inhibits a TLR7-dependent and a TLR8-dependent immune response. In some aspects, at least one TLR inhibitor is administered in an amount effective to inhibit an immune response in an individual.

[0126] Also provided herein are methods for treating or preventing an autoimmune disease in an individual, comprising administering to the individual an effective amount of a TLR7 and / or TLR8 inhibitor. In some aspects, the autoimmune disease is characterized by joint pain, positive antinuclear antibodies, a malar rash, or a discoid rash. In some aspects, the autoimmune disease is associated with skin, muscle tissue, and / or connective tissue. In some embodiments, the autoimmune disease is not apparent in the individual from symptoms in the skin, muscle tissue, and / or connective tissue. In some embodiments, the autoimmune disease is systemic. Autoimmune diseases include, but are not limited to, rheumatoid arthritis (RA), autoimmune pancreatitis (AIP), systemic lupus erythematosus (SLE), type 1 diabetes (diabetes mellitus), multiple sclerosis (MS), antiphospholipid syndrome (APS), sclerosing cholangitis, systemic onset arthritis, irritable bowel disease (IBD), scleroderma, Sjogren's disease, vitiligo, polymyositis, pemphigus vulgaris, pemphigus foliaceus, inflammatory bowel disease including Crohn's disease and ulcerative colitis, autoimmune hepatitis, hypopituitarism, graft-versus-host disease (GvHD), autoimmune skin diseases, uveitis, pernicious anemia, and hypoparathyroidism. Autoimmune diseases may also include, but are not limited to, polyangiitis overlap syndrome, Kawasaki disease, sarcoidosis, glomerulonephritis, and cold sores.

[0127] In some aspects, the autoimmune disease is selected from the group consisting of arthritis, pancreatitis, mixed connective tissue disease (MCTD), lupus, antiphospholipid syndrome (APS), systemic arthritis, and irritable bowel syndrome.

[0128] In other aspects, the autoimmune disease is selected from the group consisting of systemic lupus erythematosus (SLE), rheumatoid arthritis, autoimmune skin diseases, and multiple sclerosis.

[0129] In other aspects, the autoimmune disease is selected from the group consisting of pancreatitis, glomerulonephritis, pyelitis, sclerosing cholangitis, and type I diabetes. In some aspects, the autoimmune disease is rheumatoid arthritis. In some aspects, the autoimmune disease is autoimmune pancreatitis (AIP). In some aspects, the autoimmune disease is glomerulonephritis. In some aspects, the autoimmune disease is pyelitis. In some aspects, the autoimmune disease is sclerosing cholangitis. In some aspects, the autoimmune disorder is psoriasis. In some aspects, the autoimmune disease is a rheumatoid disease or disorder. In some aspects, the rheumatoid disease or disorder is rheumatoid arthritis. In some aspects, the disease is diabetes and / or a diabetes-related disease or disorder. In some aspects, wherein the autoimmune disease is associated with RNA-containing immune complexes. In some aspects, the autoimmune disease is Sjogren's disease.

[0130] Provided herein are methods of inhibiting an immune response in an individual, the methods comprising administering to the individual at least one TLR inhibitor as disclosed herein in an amount effective to inhibit the immune response in the individual. In some variations, the immune response is associated with an inflammatory disorder. As used herein, the term "inflammatory disorder" encompasses inflammatory conditions without a known autoimmune component (e.g., atherosclerosis, asthma, etc.). In a further aspect, inhibiting the immune response ameliorates one or more symptoms of an inflammatory disorder. In yet a further aspect, inhibiting the immune response treats an inflammatory disorder. In an even further aspect, inhibiting the immune response prevents or delays the onset of an inflammatory disorder. In some aspects, the inflammatory disorder is selected from the group consisting of non-rheumatoid arthritis, renal fibrosis, and hepatic fibrosis. In some aspects, the inflammatory disorder is interface dermatitis. In some further aspects, the interface dermatitis is selected from the group consisting of lichen planus, lichenoid eruption, lichen planus-like keratosis, linear lichen, chronic lichenoid keratosis, erythema multiforme, fixed drug eruption, pityriasis lichenoides, phototoxic dermatitis, radiodermatitis, viral exanthema, dermatomyositis, secondary syphilis, lichen sclerosus et atrophicus, mycosis fungoides, bullous pemphigoid, lichen xanthoides, porokeratosis, acrodermatitis chronica atrophica, and regressing melanoma. In some aspects, the inflammatory condition is a skin disorder such as atopic dermatitis (eczema). In some aspects, the inflammatory disorder is a sterile inflammatory condition such as drug-induced liver and / or pancreatic inflammation. In some further aspects, the inflammatory disease is an inflammatory liver disorder. In some other further aspects, the inflammatory disease is an inflammatory pancreatic disorder.

[0131] Provided herein are methods of inhibiting an immune response in an individual, the methods comprising administering to the individual at least one TLR inhibitor as disclosed herein in an amount effective to inhibit the immune response in the individual. In some variations, the immune response is associated with chronic pathogen stimulation. In some variations, the immune response is associated with infection by HIV. In a further aspect, wherein inhibiting the immune response ameliorates one or more symptoms of a viral disease or disorder resulting from infection by HIV. In yet a further aspect, wherein inhibiting the immune response treats a viral disease or disorder resulting from infection by HIV. In an even further aspect, wherein inhibiting the immune response prevents or delays the onset of a viral disease or disorder resulting from infection by HIV. Other variations provided herein relate to immunosuppressive treatment of individuals exposed to or infected with HIV. Administration of a TLR inhibitor to an individual exposed to or infected with HIV results in suppression of HIV-induced cytokine production. In some aspects, at least one TLR inhibitor is administered in an amount effective to suppress HIV-induced cytokine production in an individual exposed to or infected with HIV.

[0132] Provided herein are methods for inhibiting a TLR7- and / or TLR8-dependent immune response in an individual, the method comprising administering to the individual a TLR inhibitor in an amount effective to inhibit the immune response in the individual. In some variations, the immune response is associated with an autoimmune disease. In some aspects, the autoimmune disease is rheumatoid arthritis. In some aspects, the TLR inhibitor is effective in suppressing one or more symptoms of rheumatoid arthritis. In some aspects, the autoimmune disease is multiple sclerosis. In some aspects, the TLR inhibitor is effective in suppressing one or more symptoms of multiple sclerosis. In some aspects, the autoimmune disease is lupus. In some aspects, the TLR inhibitor is effective in suppressing one or more symptoms of lupus. In some aspects, the autoimmune disease is pancreatitis. In some aspects, the TLR inhibitor is effective in suppressing one or more symptoms of pancreatitis. In some aspects, the autoimmune disease is diabetes. In some aspects, the TLR inhibitor is effective in suppressing one or more symptoms of diabetes. In some aspects, the disease is Sjogren's disease. In some aspects, the TLR inhibitor is effective in suppressing one or more symptoms of Sjogren's disease. In some variations, the immune response is associated with an inflammatory disorder. In some aspects, the TLR inhibitor is effective in suppressing one or more symptoms of an inflammatory disorder. In some variations, the immune response is associated with chronic pathogen stimulation. In some aspects, the TLR inhibitor is effective in suppressing one or more symptoms of chronic pathogen stimulation. In some variations, the immune response is associated with a viral disease resulting from infection with HIV. In some aspects, the TLR inhibitor is effective in suppressing one or more symptoms of a viral disease resulting from infection with HIV. In any variation, the TLR inhibitor is a polynucleotide comprising an inhibitory motif for one or more of TLR7, TLR8, and TLR9.

[0133] In some embodiments of any of the methods involving administration of a TLR inhibitor to an individual (e.g., methods of inhibiting an immune response, treating or preventing an autoimmune disease or inflammatory disorder, etc.), the TLR inhibitor has a therapeutically acceptable safety profile. The TLR inhibitor has a therapeutically acceptable histological profile, including, for example, acceptably low (if any) toxicity to the liver, kidney, pancreas, or other organs. Occasionally, polynucleotides are associated with toxicity to certain organs, such as the liver, kidney, and pancreas. In some embodiments, the TLR inhibitor has an unexpected and advantageous safety profile. In some embodiments, the safety profile includes assessment of toxicity, histological profile, and / or necrosis (e.g., liver, kidney, and / or heart). In some embodiments, the TLR inhibitor has a therapeutically acceptable level of toxicity. In some embodiments, the TLR inhibitor has a reduced level of toxicity compared to other TLR inhibitors. In some embodiments, the TLR inhibitor induces a therapeutically acceptable reduction in body weight compared to the treated individual's initial body weight. In some embodiments, the TLR inhibitor induces a reduction in total body weight of less than 5%, 7.5%, 10%, 12.5, or 15%. In some embodiments, the TLR inhibitor has a therapeutically acceptable histology profile. In some embodiments, the TLR inhibitor has a better (e.g., lower severity score) histology profile, e.g., compared to a reference TLR inhibitor. In some embodiments, the TLR inhibitor has a better (e.g., lower severity score) histology profile, e.g., upon evaluation of the liver, kidney, and / or heart. In some embodiments, the TLR inhibitor has a therapeutically acceptable necrosis score. In some embodiments, the TLR inhibitor has reduced necrosis and / or a better (e.g., lower) necrosis score, e.g., compared to a reference TLR inhibitor.In some embodiments, the TLR inhibitor has a reduced renal and / or hepatic cell necrosis score and / or a better renal and / or hepatic cell necrosis score, eg, compared to a reference TLR inhibitor.

[0134] Accordingly, the present invention provides a method for activating TLR7 in an animal, particularly a mammal, preferably a human, comprising administering to the animal an effective amount of a compound of Formula I. As with all compositions for inhibiting an immune response, the effective amount of a particular TLR inhibitor formulation and its method of administration can vary based on the individual, the condition being treated, and other factors apparent to one of skill in the art. Effective amounts of the compounds will vary according to factors known in the art, but are expected to be doses of about 0.1-10 mg / kg, 0.5-10 mg / kg, 1-10 mg / kg, 0.1-20 mg / kg, 0.1-20 mg / kg, or 1-20 mg / kg.

[0135] The present invention also provides a method for treating a viral infection in an animal, comprising administering to the animal an effective amount of a compound of Formula I. An amount effective to treat or inhibit a viral infection is an amount that will cause a reduction in one or more symptoms of the viral infection, such as viral lesions, viral load, viral production rate, and mortality, compared to untreated control animals. The exact amount will vary according to factors known in the art, but is expected to be a dose as indicated above for activating TLR7, or from about 100 ng / kg to about 50 mg / kg, preferably from about 10 μg / kg to about 5 mg / kg.

[0136] In various embodiments, compounds of Formula (I) and related formulae have an IC for binding to TLR7 / 8 of less than about 5 μM, preferably less than about 1 μM, and even more preferably less than about 0.100 μM. 50 It presents.

[0137] The methods of the present invention can be carried out either in vitro or in vivo. The sensitivity of specific cells to treatment with the compounds of the present invention can be specifically determined by in vitro testing, whether in a research process or for clinical application. Typically, a culture of cells is combined with a compound of the present invention at various concentrations for a period of time sufficient for the active agent to inhibit TLR7 / 8 activity, usually between about one hour and one week. In vitro treatment can be carried out using cultured cells from a biopsy sample or a cell line.

[0138] The host or patient may belong to any mammalian species, such as a primate species, particularly humans; rodents, including mice, rats, and hamsters; rabbits; horses, cows, dogs, cats, etc. Animal models are the subject of experimental investigation and provide models for the treatment of human diseases.

[0139] For the identification of signal transduction pathways and for the detection of interactions between various signal transduction pathways, various scientists have developed suitable models or model systems, such as cell culture models and transgenic animal models. For the determination of certain stages of the signal transduction cascade, interacting compounds can be used to modulate the signal. The compounds according to the invention may also be used as reagents to study TLR7 / 8-dependent signaling pathways in animal and / or cell culture models or in the clinical diseases mentioned in this application.

[0140] Moreover, the following teachings herein regarding the use of compounds according to formula (I) and derivatives thereof for the production of medicaments for prophylactic or therapeutic treatment and / or monitoring are believed to be valid and applicable, without limitation, to the use of the compounds for the inhibition of TLR7 / 8 activity.

[0141] The present invention also relates to the use of compounds according to formula (I) and / or physiologically acceptable salts thereof for the prophylactic or therapeutic treatment and / or monitoring of diseases caused, mediated, and / or propagated by TLR7 / 8 activity. Furthermore, the present invention relates to the use of compounds according to formula (I) and / or physiologically acceptable salts thereof for the production of a medicament for the prophylactic or therapeutic treatment and / or monitoring of diseases caused, mediated, and / or propagated by TLR7 / 8 activity. In one embodiment, the present invention provides the use of a compound according to formula I, or a physiologically acceptable salt thereof, for the production of a medicament for the prophylactic or therapeutic treatment of a TLR7 / 8-mediated disorder.

[0142] The compounds of formula (I) and / or their physiologically acceptable salts may also be employed as intermediates for the preparation of further pharmaceutically active ingredients. The medicaments are preferably prepared in a non-chemical manner, for example by combining the active ingredient with at least one solid, liquid and / or semi-liquid carrier or excipient, optionally in combination with one or more other active substances in a suitable dosage form.

[0143] The compounds of formula (I) according to the present invention can act as a treatment by being administered once or several times before or after the onset of a disease. The aforementioned compounds and pharmaceuticals of use in the present invention are specifically used for therapeutic treatment. A therapeutically relevant effect is to alleviate one or more symptoms of a disorder to some extent, or to partially or completely restore one or more physiological or biochemical parameters associated with or causing a disease or pathological condition to normal. Monitoring is considered a type of treatment, provided that the compounds are administered at distinct intervals, for example, to boost the response and completely eliminate the pathogens and / or symptoms of the disease. Either the same compound or different compounds can be applied. The methods of the present invention can also be used to reduce the likelihood of developing a disorder, or even to prevent the occurrence of a disorder associated with TLR7 / 8 activity in advance, or to treat existing and ongoing symptoms.

[0144] In the sense of the present invention, preventive treatment is advisable if the subject possesses any prerequisite for the aforementioned physiological or pathological conditions, such as a familial disposition, a genetic defect, or a pre-existing disease.

[0145] The present invention also relates to a medicament comprising at least one compound according to the invention and / or its pharmaceutically usable derivatives, salts, solvates and stereoisomers (including mixtures thereof in any ratio). In one embodiment, the present invention relates to a medicament comprising at least one compound according to the invention and / or its physiologically acceptable salt.

[0146] A "medicine" in the sense of the present invention is any agent in the medical field that contains one or more compounds of formula (I) or preparations thereof (e.g. pharmaceutical compositions or pharmaceutical formulations) and that can be used in the prophylaxis, treatment, monitoring or aftercare of patients suffering from diseases associated with TLR7 / 8 activity, so that a pathogenic modification of their general condition or of the condition of a specific area of ​​the organism can only be established, at least temporarily.

[0147] In various embodiments, the active ingredient may be administered alone or in combination with other treatments. A synergistic effect may be achieved by using more than one compound in a pharmaceutical composition, i.e., the compound of formula (I) is combined with at least one other agent as an active ingredient (either another compound of formula (I) or a compound of a different structural framework). The active ingredients may be used simultaneously or sequentially.

[0148] The TLR inhibitor of the present disclosure can be administered in combination with one or more additional therapeutic agents.As described herein, the TLR inhibitor can be combined with a physiologically acceptable carrier.The methods described herein can also be practiced in combination with other treatments that complement standard therapy for disorders, such as the administration of anti-inflammatory agents.

[0149] In some embodiments, the TLR inhibitor as described herein is administered in combination with a corticosteroid. In some embodiments, the corticosteroid is a glucocorticosteroid. In some embodiments, the corticosteroid is a mineralocorticoid. Corticosteroids include corticosterone and derivatives, their prodrugs, isomers and analogs, cortisone and derivatives, their prodrugs, isomers and analogs (i.e., Cortone), aldosterone and derivatives, their prodrugs, isomers and analogs, dexamethasone and derivatives, their prodrugs, isomers and analogs (i.e., Decadron), prednisone and derivatives, their prodrugs, isomers and analogs (i.e., Prelone), fludrocortisone and derivatives, their prodrugs, isomers and analogs, hydrocortisone and derivatives, their prodrugs, isomers and analogs (i.e., Cortisol or Cortef), hydroxycortisone and derivatives, their prodrugs, isomers and analogs, betamethasone and derivatives, their prodrugs, isomers and analogs (i.e., Celestone), budesonide and derivatives, their prodrugs, isomers and analogs (i.e., Entocort Corticosteroids include, but are not limited to, methylprednisolone and derivatives, their prodrugs, isomers, and analogs (i.e., Medrol), prednisolone and derivatives, their prodrugs, isomers, and analogs (i.e., Deltasone, Crtan, Meticorten, Orasone, or Sterapred), triamcinolone and derivatives, their prodrugs, isomers, and analogs (i.e., Kenacort or Kenalog), and the like. In some embodiments, the corticosteroid is fludrocortisone or a derivative, a prodrug, isomer, or analog thereof. In some embodiments, the corticosteroid is fludrocortisone. In some embodiments, the corticosteroid is hydroxycortisone or a derivative, a prodrug, isomer, or analog thereof. In some embodiments, the corticosteroid is hydroxycortisone.

[0150] In some embodiments, the corticosteroid is administered at about 0.001 mg to 1 mg, 0.5 mg to 1 mg, 1 mg to 2 mg, 2 mg to 20 mg, 20 mg to 40 mg, 40 mg to 80 mg, 80 mg to 120 mg, 120 mg to 200 mg, 200 mg to 500 mg, or 500 mg to 1000 mg per day. In some embodiments, the corticosteroid is administered at about 0.1 mg / kg to 0.5 mg / kg, 0.5 mg / kg to 1 mg / kg, 1 mg / kg to 2 mg / kg, 2 mg / kg to 5 mg / kg, 5 mg / kg to 10 mg / kg, 10 mg / kg to 15 mg / kg, 15 mg / kg to 20 mg / kg, 20 mg / kg to 25 mg / kg, 25 mg / kg to 35 mg / kg, or 35 mg / kg to 50 mg / kg per day.

[0151] In some embodiments, the TLR inhibitor used in the combination therapy is given in an amount of TLR inhibitor delivered, which may be, for example, about 0.1-10 mg / kg, 0.5-10 mg / kg, 1-10 mg / kg, 0.1-20 mg / kg, 0.1-20 mg / kg, or 1-20 mg / kg.

[0152] In some embodiments, the TLR inhibitor is administered simultaneously with one or more additional therapeutic agents, including but not limited to, corticosteroids (coadministration). In some embodiments, the TLR inhibitor is administered sequentially with one or more additional therapeutic agents, including but not limited to, corticosteroids (sequential administration). In some embodiments, sequential administration includes administering the TLR inhibitor or additional therapeutic agent within about 1 minute, 5 minutes, 30 minutes, 1 hour, 5 hours, 24 hours, 48 ​​hours, or 1 week of each other. In some embodiments, the TLR inhibitor is administered by the same route of administration as the additional therapeutic agent. In some embodiments, the TLR inhibitor is administered by a different route of administration than the additional therapeutic agent. In some embodiments, the additional therapeutic agent is administered parenterally (e.g., via a central venous line, intraarterially, intravenously, intramuscularly, intraperitoneally, intradermally, or subcutaneously), orally, gastrointestinally, topically, nasopharyngeally, and pulmonary (e.g., by inhalation or intranasally). In some embodiments, the additional therapeutic agent is a corticosteroid.

[0153] The disclosed compounds of Formula I can be administered in combination with other known therapeutic agents, including anti-cancer agents. As used herein, the term "anti-cancer agent" refers to any agent administered to a patient with cancer for the purpose of treating the cancer.

[0154] The anti-cancer treatment defined above may be applied as a monotherapy or may involve, in addition to the compound of formula I disclosed herein, conventional surgery or radiation therapy or medicinal therapy. Such medicinal therapy (e.g., chemotherapy or targeted therapy) may include one or more, preferably one of the following anti-tumor agents: Alkylating agents:Altretamine, bendamustine, busulfan, carmustine, chlorambucil, chlormethine, cyclophosphamide, dacarbazine, ifosfamide, improsulfan, tosilate, lomustine, melphalan, mitobronitol, mitolactol, nimustine, ranimustine, temozolomide, thiotepa, treosulfan, mechlorethamine, carboquone; apaziquone, fotemustine, glufosfamide, palifosfamide, pipobroman, trofosfamide, uramustine, TH-302 4 , VAL-083 4 etc; Platinum compounds: Carboplatin, cisplatin, eptaplatin, miriplatin hydrate, oxaliplatin, lobaplatin, nedaplatin, picoplatin, satraplatin; lobaplatin, nedaplatin, picoplatin, satraplatin, etc.; DNA modifying agents: Amrubicin, bisantrene, decitabine, mitoxantrone, procarbazine, trabectedin, clofarabine; amsacrine, brostallicin, pixantrone, laromustine 1,3 etc; Topoisomerase inhibitors: Etoposide, irinotecan, razoxane, sobuzoxane, teniposide, topotecan; amonafide, belotecan, elliptinium acetate, voreloxin, etc.; Microtubule modifier: Cabazitaxel, docetaxel, eribulin, ixabepilone, paclitaxel, vinplastine, vincristine, vinorelbine, vindesine, vinflunine; fosbretabine, tesetaxel, etc. Antimetabolites: Asparaginase 3 , azacitidine, levofolinate calcium, capecitabine, cladribine, cytarabine, enocitabine, floxuridine, fludarabine, fluorouracil, gemcitabine, mercaptopurine, methotrexate, nelarabine, pemetrexed, pralatrexate, azathioprine, thioguanine, carmofur; doxifluridine, ellacitabine, raltitrexed, cepacitabine, tegafur2,3 , trimethotrexate, etc.; Anticancer antibiotics: Bleomycin, dactinomycin, doxorubicin, epirubicin, idarubicin, levamisole, miltefosine, mitomycin C, romidepsin, streptozocin, valrubicin, zinostatin, zorubicin, daunorubicin, plicamycin; aclarubicin, peplomycin, pirarubicin, etc.; Hormones / antagonists: Abarelix, abiraterone, bicalutamide, buserelin, calcitonin, chlorothiazol-1, degarelix, dexamethasone, estradiol, flutocortolone, fluoxymesterone, flutamide, fulvestrant, goserelin, histrelin, leuprorelin, megesterol, mitotane, nafarelin, nandrolone, nilutamide, octreotide, prednisolone, raloxifene, tamoxifen, thyrotropin alfa, toremifene, trilostane, triptorelin, diethylstilbestrol; acolbifene, danazol, deslorelin, epithiostanol, orteronel, enzalutamide 1,3 etc; Aromatase inhibitors: Aminoglutethimide, anastrozole, exemestane, fadrozole, letrozole, testolactone; formestane, etc.; Small molecule kinase inhibitors: Crizotinib, dasatinib, erlotinib, imatinib, lapatinib, nilotinib, pazopanib, regorafenib, ruxolitinib, sorafenib, sunitinib, vandetanib, vemurafenib, bosutinib, gefitinib, axitinib; afatinib, alisertib, dabrafenib, dacomitinib, dinaciclib, dovitinib, etanercept Inzastaurin, nintedanib, lenvatinib, linifanib, linsitinib, masitinib, midostaurin, motesanib, neratinib, orantinib, perifosine, ponatinib, radotinib, rigosertib, tipifanib, tivantinib, tivozanib, trametinib, pimasertib, brivanib alaninate, cediranib, apatinib 4 , Cabozantinib S-malate 1,3 , ibrutinib 1,3 , icotinib4 , buparlisib 2 , cipatinib 4 , cobimetinib 1,3 , idelalisib 1,3 , fedratinib 1 , XL-647 4 etc; Photosensitizers: Methoxsalen 3 ;Porfimer sodium, talaporfin, temoporfin, etc.; antibody: Alemtuzumab, besilesomab, brentuximab vedotin, cetuximab, denosumab, ipilimumab, ofatumumab, panitumumab, rituximab, tositumomab, trastuzumab, bevacizumab, pertuzumab 2,3 Catumaxomab, elotuzumab, epratuzumab, farletuzumab, mogamulizumab, necitumumab, nimotuzumab, obinutuzumab, ocalatuzumab, oregovomab, ramucirumab, rilotumumab, siltuximab, tocilizumab, zalutumumab, zanolimumab, matuzumab, dalotuzumab 1,2,3 , onartuzumab 1,3 , racotumomab 1 , tabalumab 1,3 , EMD-525797 4 , nivolumab 1,3 etc; Cytokines: Aldesleukin, interferon alfa 2 , interferon alpha 2a 3 , interferon alfa-2b 2,3 ; Celmoleukin, tasonermin, teseleukin, operelvekin 1,3 , recombinant interferon beta-1a 4 etc; Drug conjugates:Denileukin diftitox, ibritumomab tiuxetan, iobenguane I123, prednimustine, trastuzumab emtansine, estramustine, gemtuzumab, ozogamicin, aflibercept; syntredequin besudotox, edotreotide, inotuzumab ozogamicin, naptumomab estafenatox, oportuzumab monatox, technitium (99mTc) arcitumomab 1,3 , vintaphorid 1,3 etc; vaccine: Sipuleucel 3 ;Vitespen 3 , Emepepimt-S 3 , OncoVAX 4 , Lindopepimt 3 , troVax 4 , MGN-1601 4 , MGN-1703 4 etc.; and others: Alitretinoin, bexarotene, bortezomib, everolimus, ibandronate, imiquimod, lenalidomide, lentinan, metyrosine, mifamurtide, pamidronate, pegaspargase, pentostatin, sipuleucel 3 , sizofiran, tamibarotene, temsirolimus, thalidomide, tretinoin, vismodegib, zoledronic acid, vorinostat; celecoxib, cilengitide, entinostat, etanidazole, ganetespib, idronoxyl, iniparib, ixazomib, lonidamine, nimorazole, parabinostat, peretinoin, plitidepsin, pomalidomide, procodazol, ridaforolimus, tasquinimod, telotristat, simalfasin, tirapazamine, tosedostat, travedelsen, ubenimex, valspodar, and gendicine 4 , Picibanil 4 , leiolysin 4 , letaspimycin hydrochloride 1,3 , trebananib 2,3 , bilirudin 4 , carfilzomib 1,3 , endostatin 4, Immucothel 4 , Belinostat 3 , MGN-1703 4 . ( 1 Prop. INN (Proposed International Nonproprietary Name); 2 Rec. INN (Recommended International Nonproprietary Name); 3 USAN (United States Adopted Name); 4 No INN).

[0155] In some embodiments, the combination of a TLR inhibitor and one or more additional therapeutic agents reduces the effective amount (including, but not limited to, dosage volume, dosage concentration, and / or total drug dose) of the TLR inhibitor and / or one or more additional therapeutic agents administered to achieve the same result, compared to the effective amount administered when the TLR inhibitor or additional therapeutic agent is administered alone. In some embodiments, the combination of a TLR inhibitor and a corticosteroid reduces the effective amount of the corticosteroid administered, compared to the corticosteroid administered alone. In some embodiments, the combination of a TLR inhibitor and an additional therapeutic agent reduces the frequency of therapeutic agent administration, compared to administration of the additional therapeutic agent alone. In some embodiments, the combination of a TLR inhibitor and an additional therapeutic agent reduces the total treatment duration, compared to administration of the additional therapeutic agent alone. In some embodiments, the combination of a TLR inhibitor and an additional therapeutic agent reduces side effects associated with administration of the additional therapeutic agent alone. In some embodiments, the additional therapeutic agent is a corticosteroid. In some embodiments, the corticosteroid is fludrocortisone or a derivative, prodrug, isomer, or analog thereof. In some embodiments, the corticosteroid is fludrocortisone. In some embodiments, the combination of an effective amount of a TLR inhibitor and an additional therapeutic agent is more effective than an effective amount of either the TLR inhibitor or the additional therapeutic agent alone.

[0156] TLR inhibitors may also be useful as vaccine adjuvants for use in conjunction with any material that modulates either the humoral and / or cell-mediated immune response (e.g., live viral, bacterial, or parasitic immunogens; inactivated viral, tumor-derived, protozoan, organism-derived, fungal, or bacterial immunogens, toxoids, toxins; autoantigens; polysaccharides; proteins; glycoproteins; peptides; cellular vaccines; DNA vaccines; recombinant proteins; glycoproteins; peptides; etc.). In some aspects, combination therapy, including but not limited to, a combination of a TLR inhibitor and a vaccine, is used in the treatment of an autoimmune disease or inflammatory disorder. In some aspects, combination therapy, including but not limited to, a combination of a TLR inhibitor and a vaccine, is used in the treatment of an infectious disease.

[0157] In some embodiments, the combination therapy, including but not limited to the combination of TLR inhibitor and corticosteroid, is used to treat autoimmune disease or inflammatory disorder.In some embodiments, the autoimmune disease is selected from but not limited to rheumatoid arthritis, systemic lupus erythematosus, autoimmune skin disease, multiple sclerosis, pancreatitis, glomerulonephritis, pyelitis, sclerosing cholangitis and type I diabetes.In some embodiments, the autoimmune disease is Sjogren's disease.

[0158] Also provided herein are kits that include a TLR inhibitor as provided herein and instructions for use in a method of inhibiting a TLR7- and / or TLR8-dependent immune response.

[0159] The kit may include one or more containers containing a TLR inhibitor (or a formulation containing a TLR inhibitor) as described herein and a set of instructions, typically written instructions, although electronic storage media (e.g., a magnetic diskette or optical disk) containing instructions regarding the use and dosage of the TLR inhibitor or formulation for an intended treatment (e.g., suppressing responses to TLR7 and / or TLR8 agonists, suppressing TLR7- and / or TLR8-dependent immune responses, ameliorating one or more symptoms of an autoimmune disease, ameliorating symptoms of a chronic inflammatory disease, reducing cytokine production in response to a virus, and / or treating and / or preventing one or more symptoms of a disease or disorder mediated by TLR7 and / or TLR8) are also acceptable. The instructions included in the kit generally include information regarding the dosage, dosing schedule, and route of administration for the intended treatment. The containers for the TLR inhibitor (or formulations comprising the TLR inhibitor) may be unit doses, bulk packages (e.g., multi-dose packages) or sub-unit doses. The kit may further comprise a container containing an adjuvant.

[0160] In another aspect, the present invention provides a kit comprising separate packs of effective amounts of compounds according to the invention and / or their pharmaceutically acceptable salts, derivatives, solvates and stereoisomers (including mixtures thereof in any ratio), and, optionally, effective amounts of further active ingredients, the kit comprising suitable containers such as boxes, individual bottles, bags or ampoules. The kit may, for example, comprise separate ampoules, each containing an effective amount of a compound according to the invention and / or its pharmaceutically acceptable salts, derivatives, solvates and stereoisomers (including mixtures thereof in any ratio), and optionally an effective amount of a further active ingredient, in dissolved or lyophilized form.

[0161] As used herein, the terms "treatment," "treat," and "treating" refer to reversing, alleviating, delaying, or inhibiting the progression of a disease or disorder as described herein, or one or more symptoms thereof. In some embodiments, treatment is administered after one or more symptoms have developed. In other embodiments, treatment is administered in the absence of symptoms. For example, treatment is administered to a susceptible individual prior to the onset of symptoms (e.g., in light of a history of the condition and / or in light of genetic or other susceptibility factors). Treatment also continues after symptoms have resolved, e.g., to prevent or delay their recurrence.

[0162] According to the method of the present invention, the compounds and compositions are administered in any amount and using any route of administration effective to treat or reduce the severity of the disorders provided above. The exact amount required will vary from subject to subject, depending on the species, age, and general condition of the subject, the severity of the infection, the specific agent, its mode of administration, and the like. The compounds of the present invention are preferably formulated in dosage unit form for ease of administration and uniformity of dosage. As used herein, the expression "dosage unit form" refers to a physically discrete unit of agent appropriate for the patient to be treated. However, it will be understood that the total daily usage of the compounds and compositions of the present invention will be determined by the attending physician within the scope of sound medical judgment. The specific dosage level effective for any particular patient or organism will depend on a variety of factors, including the disorder being treated and the severity of the disorder; the activity of the particular compound employed; the particular compound employed; the patient's age, weight, general health, sex, and diet; the time of administration, route of administration, and rate of excretion of the particular compound employed; the duration of treatment; drugs used in combination or concomitantly with the particular compound employed, and similar factors well known in the medical arts.

[0163] The pharmaceutically acceptable compositions of the present invention can be administered to humans and other animals orally, rectally, parenterally, intracisternally, intravaginally, intraperitoneally, topically (such as by powder, ointment, or drops), buccally, as an oral or nasal spray, or the like, depending on the severity of the infection being treated. In some embodiments, the compounds of the present invention are administered orally or parenterally at dosage levels of from about 0.01 mg / kg to about 100 mg / kg of body weight (of the subject) per day, preferably from about 1 mg / kg to about 50 mg / kg of body weight (of the subject), one or more times per day to achieve the desired therapeutic effect.

[0164] In certain embodiments, the therapeutically effective amount of a compound of Formula (I) and related formulas, as well as the amount of other active ingredients, will depend on numerous factors, including, for example, the age and weight of the animal, the exact disease state and its severity, the nature of the formulation, and the method of administration, and will ultimately be determined by the treating physician or veterinarian. However, an effective amount of a compound will generally range from 0.1 to 100 mg / kg body weight (of the recipient (mammal)) per day, and more typically from 1 to 10 mg / kg body weight per day. Thus, the actual daily dose for an adult mammal weighing 70 kg will usually be between 70 mg and 700 mg, although this amount can be administered as individual doses per day, or in a set of partial doses (e.g., 2, 3, 4, 5, 6, etc.) per day so that the total daily dose is usually the same. An effective amount of a salt or solvate, or an effective amount of a physiologically functional derivative thereof, can be determined as a fraction of the effective amount of the compound itself.

[0165] In some embodiments, pharmaceutical preparations can be administered in the form of dosage units, each containing a predetermined amount of active ingredient. Such units can contain, for example, 0.5 mg to 1 g, preferably 1 mg to 700 mg, particularly preferably 5 mg to 100 mg of the compound according to the present invention, depending on the disease state to be treated, the method of administration, and the age, weight, and condition of the patient. Alternatively, pharmaceutical preparations can be administered in the form of dosage units, each containing a predetermined amount of active ingredient. Preferred dosage unit preparations are those containing the daily dose or partial dose indicated above, or a corresponding fraction thereof, of the active ingredient. Furthermore, pharmaceutical preparations of this type can be prepared using processes commonly known in the pharmaceutical arts.

[0166] Liquid dosage forms for oral administration include, but are not limited to, pharmaceutically acceptable emulsions, microemulsions, solutions, suspensions, syrups and elixirs. In addition to the active compound, the liquid dosage form optionally contains an inert diluent commonly used in the art, such as water or other solvents, solubilizers and emulsifiers such as ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butylene glycol, dimethylformamide, oils (especially cottonseed oil, peanut oil, corn oil, germ oil, olive oil, castor oil, and sesame oil), glycerol, tetrahydrofurfuryl alcohol, polyethylene glycol, and fatty acid esters of sorbitan, and mixtures thereof. In addition to the inert diluent, the oral composition also contains adjuvants such as wetting agents, emulsifying agents, and suspending agents, sweeteners, flavoring agents, and coloring agents.

[0167] Injectable preparations, for example, sterile injectable aqueous or oleaginous suspensions, are formulated according to known techniques using suitable dispersants or wetting agents and suspending agents.Sterile injectable preparations can also be sterile injectable solutions, suspensions, or emulsions in non-toxic parenterally acceptable diluents or solvents, such as solutions in 1,3-butanediol.Among acceptable vehicles and solvents that can be used are water, Ringer's solution, USP, and isotonic sodium chloride solution.In addition, sterile fixed oils are conventionally used as solvents or suspending media.For this purpose, any bland fixed oil that can be used includes synthetic mono- or diglycerides.In addition, fatty acids such as oleic acid are used in injectable preparations.

[0168] Injectable preparations can be sterilized, for example, by filtration through a bacterial-retaining filter, or by incorporating sterilizing agents in the form of sterile solid compositions which can be dissolved or dispersed in sterile water or other sterile injectable medium prior to use.

[0169] To prolong the effect of a compound of the present invention, it is often desirable to slow the absorption of the compound from subcutaneous or intramuscular injection. This can be accomplished by using a liquid suspension of poorly soluble crystalline or amorphous material. The rate of absorption of the compound then depends on its dissolution rate, which in turn may depend on crystal size and crystalline form. Alternatively, delayed absorption of a parenterally administered compound form can be achieved by dissolving or suspending the compound in an oil vehicle. Injectable depot forms are made by forming microencapsulated matrices of the compound in biodegradable polymers such as polylactide-polyglycolide. Depending on the compound-to-polymer ratio and the nature of the specific polymer employed, the compound release rate can be controlled. Examples of other biodegradable polymers include poly(orthoesters) and poly(anhydrides). Injectable depot formulations can also be prepared by entrapping the compound in liposomes or microemulsions that are compatible with body tissues.

[0170] Compositions for rectal or vaginal administration are preferably suppositories which may be prepared by mixing a compound of the invention with a suitable non-irritating excipient or carrier, such as cocoa butter, polyethylene glycol, and the like, or a suppository wax which is solid at ambient temperature but liquid at body temperature and which melts in the rectum or vaginal cavity to release the active compound.

[0171] Solid dosage forms for oral administration include capsules, tablets, pills, powders, and granules. In such solid dosage forms, the active compound is incorporated into at least one pharmaceutically acceptable excipient or carrier, such as sodium citrate or dicalcium phosphate, and / or a) fillers or extenders, such as starch, lactose, sucrose, glucose, mannitol, and silicic acid; b) binders, such as, for example, carboxymethylcellulose, alginic acid, gelatin, polyvinylpyrrolidinone, sucrose, and gum acacia; c) humectants, such as glycerol; d) agar, calcium carbonate, potato or tapioca starch, alginic acid, They are mixed with disintegrating agents such as certain silicates and sodium carbonate, e) solution retarding agents such as paraffin, f) absorption accelerators such as quaternary ammonium compounds, g) wetting agents such as cetyl alcohol and glycerol monostearate, h) absorbents such as kaolin and bentonite clay, and i) lubricants such as talc, calcium stearate, magnesium stearate, solid polyethylene glycol, sodium lauryl sulfate, and mixtures thereof. In the case of capsules, tablets, and pills, the dosage form also optionally comprises buffering agents.

[0172] Solid compositions of a similar type are also employed as fillers in soft and hard-filled gelatin capsules using excipients such as lactose or milk sugar and high molecular weight polyethylene glycols. Solid dosage forms such as tablets, dragees, capsules, pills, and granules can be prepared with coatings and shells, such as enteric coatings and other coatings well known in the pharmaceutical formulation art. They can optionally contain opacifying agents and can also be of a composition that they release the active ingredient(s) only, or preferentially, in a certain part of the intestinal tract, optionally in a delayed manner. Examples of embedding compositions that can be used include polymeric substances and waxes. Solid compositions of a similar type are also employed as fillers in soft and hard-filled gelatin capsules using excipients such as lactose or milk sugar and high molecular weight polyethylene glycols.

[0173] The active compound may also be in microencapsulated form with one or more excipients as noted above. Solid dosage forms such as tablets, dragees, capsules, pills, and granules may be prepared with coatings and shells, such as enteric coatings, release-controlling coatings, and other coatings well known in the pharmaceutical formulation art. In such solid dosage forms, the active compound may be admixed with at least one inert diluent, such as sucrose, lactose, or starch. Conventional dosage forms also contain additional substances other than inert diluents, such as tableting lubricants and other tableting aids, such as magnesium stearate and microcrystalline cellulose. In the case of capsules, tablets, and pills, the dosage forms also optionally contain buffering agents. They may optionally contain opacifying agents and may be of a composition that they release the active ingredient(s) only, or preferentially, in a certain part of the intestinal tract, optionally in a delayed manner. Examples of embedding compositions that can be used include polymeric substances and polymeric waxes.

[0174] Dosage forms for topical or transdermal administration of a compound of the present invention include ointments, pastes, creams, lotions, gels, powders, solutions, sprays, inhalants, or patches. The active ingredient is admixed under sterile conditions with a pharmaceutically acceptable carrier and any necessary preservatives or buffers. Ophthalmic formulations, ear drops, and eye drops are also contemplated as being within the scope of the present invention. In addition, the present invention contemplates the use of transdermal patches, which have the added advantage of providing controlled delivery of the compound to the body. Such dosage forms can be made by dissolving or dispersing the compound in the proper medium. Absorption enhancers can also be used to increase the flux of the compound across the skin. The rate can be controlled by either providing a rate-controlling membrane or dispersing the compound in a polymer matrix or gel.

[0175] According to one aspect, the present invention relates to a method of inhibiting TLR7 / 8 activity in a biological sample, said method comprising the step of contacting said biological sample with a compound of the present invention or a composition comprising said compound.

[0176] According to another aspect, the present invention relates to a method for actively inhibiting TLR7 / 8, or a mutant thereof, activity in a biological sample, said method comprising the step of contacting said biological sample with a compound of the present invention or a composition comprising said compound.

[0177] The compounds of the present invention are useful in vitro as unique tools for understanding the biological role of TLR7 / 8, including the evaluation of numerous factors thought to affect and be affected by TLR7 / 8 production and TLR7 / 8 interaction. The compounds are also useful for the development of other compounds that interact with TLR7 / 8, because they provide important structure-activity relationship (SAR) information that facilitates their development. Compounds of the present invention that bind to TLR7 / 8 can be used as reagents for detecting TLR7 / 8 from live cells, fixed cells, biological fluids, tissue homogenates, purified natural biological materials, and the like. For example, by labeling such compounds, cells expressing TLR7 / 8 can be identified. In addition, based on their ability to bind to TLR7 / 8, the compounds of the present invention can be used in enzyme purification or to purify cells expressing TLR7 / 8 inside permeabilized cells, such as in-situ staining, FACS (fluorescence-activated cell sorting), sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE), ELISA (enzyme-linked immunosorbent assay), etc. The compounds of the present invention can also be utilized as commercial research reagents for various medical research and diagnostic uses.Such uses include, but are not limited to, the following: use as a calibration standard to quantify the activity of candidate TLR7 / 8 inhibitors in various functional assays; use as a blocking agent in random screening of compounds, i.e., in searching for new families of TLR7 / 8 ligands, where the compounds can be used to block the recovery of the presently claimed TLR7 / 8 compounds; use in co-crystallization with TLR7 / 8, i.e., the compounds of the present invention will form crystals of the compound bound to TLR7 / 8, allowing the determination of the enzyme / compound structure by X-ray crystallography; other research and diagnostic applications, where TLR7 / 8 is preferably activated, or such activation is conveniently calibrated against a known quantity, such as a TLR7 / 8 inhibitor; use in assays as a probe to determine the expression of TLR7 / 8 in cells; and developing assays to detect compounds that bind to the same site as TLR7 / 8-binding ligands.

[0178] The compounds of the present invention can be applied either by themselves and / or in combination with physical measurements for the diagnosis of treatment efficacy. Pharmaceutical compositions containing the compounds and their use to treat TLR7 / 8-mediated conditions are promising new approaches to broad-spectrum treatments that cause direct and immediate improvement in the state of health, whether in humans or animals. The new orally bioavailable and active chemical entities of the present invention improve convenience for patients and compliance for physicians.

[0179] The compounds of formula (I), their salts, isomers, tautomers, enantiomeric forms, diastereomers, racemates, derivatives, prodrugs and / or metabolites are characterized by high specificity and stability, low production costs and convenient handling. These features form the basis for a reproducible action (which includes the lack of cross-reactivity) and for a reliable and safe interaction with target structures.

[0180] The term "biological sample," as used herein, includes, without limitation, cell cultures or extracts thereof; biopsies obtained from mammals or extracts thereof; and blood, saliva, urine, feces, semen, tears, or other bodily fluids, or extracts thereof.

[0181] Modulation of TLR7 / 8, or mutants thereof, activity in biological samples is useful for a variety of purposes known to those of skill in the art, including, but not limited to, blood transfusion, organ transplantation, biological specimen storage, and biological assays.

[0182] Example As depicted in the examples below, in certain exemplary embodiments, compounds are prepared according to the following general procedures: While the general methods depict the synthesis of certain compounds of the present invention, it will be understood that the following general methods, as well as other methods known to those skilled in the art, can be applied to all compounds as described herein, and each subclass and species of these compounds.

[0183] The symbols and conventions used in the described processes, schemes, and examples that follow are consistent with those used in the contemporary scientific literature, e.g., the Journal of the American Chemical Society or the Journal of Biological Chemistry.

[0184] Unless otherwise indicated, all temperatures are expressed in degrees Celsius.

[0185] All solvents used were commercially available and were used without further purification. Reactions were typically carried out using anhydrous solvents under an inert atmosphere of nitrogen. Flash column chromatography was generally performed using Silica gel 60 (0.035-0.070 mm particle size).

[0186] All NMR experiments were recorded on either a Bruker Mercury Plus 400 NMR spectrometer equipped with a Bruker 400BBFO probe at 400 MHz for proton NMR, or a Bruker Mercury Plus 300 NMR spectrometer equipped with a Bruker 300BBFO probe at 300 MHz for proton NMR, or a Bruker Avance III 400 NMR spectrometer equipped with a Bruker PABBO BB-1H / DZ GRD probe at 400 MHz for proton NMR. All deuterated solvents typically contained 0.03%–0.05% v / v tetramethylsilane, which was used as a reference signal ( 1 H and 13 (The d and C are both set to 0.00.) In cases where the deuterated solvent did not contain tetramethylsilane, the peaks of the remaining non-deuterated solvent were used as reference signals according to published guidelines (J. Org. Chem., Vol. 62, No. 21, 1997).

[0187] LC-MS analysis was performed on one of two instruments:

[0188] LC-MS analysis was performed on a SHIMADZU LC-MS instrument consisting of a UFLC 20-AD system and an LCMS 2020 MS detector. The column used was a Shim-pack XR-ODS, 2.2 μm, 3.0 × 50 mm. A linear gradient was applied, starting at 95% A (A: 0.05% TFA in water) and ending at 100% B (B: 0.05% TFA in acetonitrile) over 2.2 min. The total run time was 3.6 min. The column temperature was 40 °C with a flow rate of 1.0 mL / min. The diode array detector scanned from 200 to 400 nm. The mass spectrometer was equipped with an electrospray ion source (ES) operated in positive or negative mode. The mass spectrometer scanned from m / z 90 to 900 with a scan time of 0.6 s.

[0189] Agilent 1200 Series mass spectrometer (Agilent Technologies) using either atmospheric pressure chemical ionization (APCI) or electrospray ionization (ESI). The diode array detector was scanned from 200-400 nm. The mass spectrometer was scanned between m / z 90-900 with a scan time of 0.6 seconds. Column: XBridge C8, 3.5 μm, 4.6 x 50 mm; Solvent A: water + 0.1% TFA; Solvent B: ACN + 0.1% TFA; Flow rate: 2 ml / min; Gradient: 0 min: 5% B, 8 min: 100% B, 8.1 min: 100% B, 8.5 min: 5% B, 10 min: 5% B; or LC / MS Waters ZMD (ESI).

[0190] HPLC data were obtained either from a SHIMAZU LC-MS instrument or from an Agilent 1100 series HPLC from Agilent technologies using a column (XBridge C8, 3.5 μm, 4.6 × 50 mm) and two mobile phases (Mobile Phase A: Water + 0.1% TFA; Mobile Phase B: ACN + 0.1% TFA). The flow rate was 2 ml / min. The gradient method, unless otherwise indicated, was: 0 min: 5% B; 8 min: 100% B; 8.1 min: 100% B; 8.5 min: 5% B; 10 min: 5% B.

[0191] In general, compounds according to Formula (I) and related formulae of the present invention can be prepared from readily available starting materials. If such starting materials are not commercially available, they may be prepared by standard synthetic techniques. In general, the synthetic route for any individual compound represented by Formula (I) and related formulae will depend on the specific substituents of each molecule. Such factors are understood by those of ordinary skill in the art. The following general methods and procedures, described below in the examples, may be employed to prepare compounds represented by Formula (I) and related formulae. Reaction conditions depicted in the following schemes, such as temperatures, solvents, or co-reagents, are given by way of example only and are not intended to be limiting. Where typical or preferred experimental conditions (i.e., reaction temperature, time, moles of reagents, solvents, etc.) are given, it will be understood that other experimental conditions can also be used unless so stated. Optimum reaction conditions may vary with the specific reagents or solvents used, but such conditions can be determined by one of ordinary skill in the art using routine optimization procedures. For a complete discussion of protection and deprotection methods, see Philip J. Kocienski, in "Protecting Groups", Georg Thieme Verlag Stuttgart, New York, 1994, and Theodora W. Greene and Peter GM Wuts, in "Protective Groups in Organic Synthesis", Wiley Interscience, 3 rdSee Edition 1999. Intermediate 1: 8-Bromoquinoxaline-5-carbonitrile [ka]

[0192] 5-Bromo-8-methylquinoxaline: To a solution of 5-methylquinoxaline (9.50 g, 66.0 mmol) in acetonitrile (80 mL) was added 1-bromopyrrolidone-2,5-dione (27.0 g, 151.7 mmol) at room temperature. The resulting solution was stirred at 60 °C for 16 h. After cooling to room temperature, the reaction mixture was concentrated under reduced pressure, and the residue was diluted with ethyl acetate (500 mL). The insoluble solid in the mixture was filtered off, and the filtrate was washed with brine and dried over Na SO . The solvent was removed under reduced pressure to afford 5-bromo-8-methylquinoxaline as a brown solid (6.00 g, 41%). MS: m / z = 222.9 [M+H] + .

[0193] 5-Bromo-8-(dibromomethyl)quinoxaline: To a solution of 5-bromo-8-methylquinoxaline (6.00 g, 27.0 mmol) in CCl (200 mL) was added NBS (19.2 g, 108.1 mmol) and AIBN (0.71 g, 4.3 mmol) at room temperature. The resulting solution was then stirred at 80 °C for 16 h. After cooling to room temperature, the reaction mixture was concentrated under reduced pressure, and the residue was diluted with ethyl acetate (500 mL). The insoluble solid in the mixture was filtered off, and the filtrate was washed with brine and dried over Na SO . The solvent was removed under reduced pressure, and the residue was purified by flash chromatography eluting with EtOAc in petroleum ether (0% to 5% gradient) to afford 5-bromo-8-(dibromomethyl)quinoxaline as a pale yellow solid (7.15 g, 70%). MS: m / z = 378.7 [M+H] + .

[0194] 8-Bromoquinoxaline-5-carbaldehyde: To a solution of 5-bromo-8-(dibromomethyl)quinoxaline (13.5 g, 35.7 mmol) in ethanol (290 mL) was added dropwise a solution of AgNO3 (24.3 g, 142.9 mmol) in water (90 mL) at room temperature. The resulting mixture was then stirred at room temperature for 1 h. Upon completion of the reaction, the reaction mixture was diluted with acetonitrile (300 mL), resulting in precipitation. The precipitate was filtered off, and the filtrate was concentrated under reduced pressure to yield 8-bromoquinoxaline-5-carbaldehyde as a yellow solid (10.0 g, crude). MS: m / z = 236.8 [M+H] + .

[0195] (E)-8-Bromoquinoxaline-5-carbaldehyde oxime: To a solution of 8-bromoquinoxaline-5-carbaldehyde (10 g, crude) in ethanol (100 mL) was added NaOAc (6.34 g, 73.4 mmol) and NHOH·HCl (3.12 g, 42.7 mmol) at room temperature. The resulting mixture was stirred at 70 °C for 3 h. Upon completion of the reaction, the insoluble solid in the reaction mixture was filtered off at 70 °C, and the filtrate was cooled to 0 °C, resulting in precipitation. The precipitate was collected by filtration and dried in an oven to afford (E)-N-[(8-bromoquinoxalin-5-yl)methylidene]hydroxylamine as a yellow solid (2.96 g, 33% for two steps). MS: m / z = 253.9 [M+H] + .

[0196] 8-Bromoquinoxaline-5-carbonitrile: To a solution of (E)-N-[(8-bromoquinoxalin-5-yl)methylidene]hydroxylamine (3.47 g, 13.8 mmol) in acetonitrile (20 mL) was added Cu(OAc) (577 mg, 3.18 mmol) and acetic acid (1.24 g, 20.7 mmol) at room temperature. The resulting mixture was stirred at 88 °C for 15 h. After cooling to room temperature, the reaction mixture was diluted with acetonitrile (10 mL). The insoluble solid in the mixture was filtered off, and the filtrate was concentrated under reduced pressure. The residue was purified by flash chromatography eluting with EtOAc in petroleum ether (0% to 5% gradient) to afford 8-bromoquinoxaline-5-carbonitrile as a yellow solid (1.22 g, 38%). MS: m / z=235.8 [M+H] + . Intermediate 2: 5-Bromo-8-methyl-[1,7]naphthyridine [ka]

[0197] 5-Bromo-8-methyl-[1,7]naphthyridine: To a mixture of 5-bromo-2-methyl-pyridin-3-ylamine (3.00 g; 16.0 mmol), glycerol (4.7 mL; 64.1 mmol), and iron(II) sulfate heptahydrate (892 mg; 3.2 mmol), sulfuric acid (5.6 mL; 96.2 mmol) was added dropwise. The resulting mixture was heated at 120° C. overnight. The reaction mixture was treated with ice, 2N sodium hydroxide solution, ethyl acetate, and dichloromethane. After filtration to remove the dark brown solid, the organic layer was separated, washed with brine, dried, and concentrated. The crude product was purified by chromatography on silica gel eluting with ethyl acetate and hexane to give 5-bromo-8-methyl-[1,7]naphthyridine (470 mg, 13%). MS: m / z=224 [M+H] + . Intermediate 3: 8-chloropyrido[2,3-b]pyrazine [ka]

[0198] 8-Chloropyrido[2,3-b]pyrazine: To a solution of 4-chloropyridine-2,3-diamine (1.90 g, 13.20 mmol) in THF (100 mL) was added oxalaldehyde (1.00 g, 17.20 mmol) at room temperature. The resulting solution was then stirred at room temperature for 6 h. Upon completion of the reaction, the reaction mixture was concentrated under reduced pressure, and the residue was purified by flash chromatography eluting with EtOAc in petroleum ether (0% to 50% gradient) to afford 8-chloropyrido[2,3-b]pyrazine as a yellow solid (2.10 g, 91%). MS: m / z = 166.1 [M+H] + . Intermediate 4: 4-Bromo-1,2-dimethyl-1H-pyrrolo[2,3-b]pyridine [ka]

[0199] 4-Bromo-1,2-dimethyl-1H-pyrrolo[2,3-b]pyridine: 0 o At C, sodium hydride (1.62 g, 40.5 mmol) was added to a solution of 4-bromo-2-methyl-1H-pyrrolo[2,3-b]pyridine (6.00 g, 27.0 mmol) in N,N-dimethylformamide (60 mL). o C for 15 min, then iodomethane (2.1 mL, 32.4 mmol) was added, and the resulting mixture was stirred at room temperature for 8 h. When the reaction was complete, the reaction mixture was quenched with water (250 mL), and the resulting mixture was extracted with ethyl acetate (3 x 100 mL). The combined organic phases were washed with brine, dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residual gum was triturated with hexane (40 mL) and dried to afford 4-bromo-1,2-dimethyl-1H-pyrrolo[2,3-b]pyridine as a brown semi-solid (3.80 g, 62%). MS: m / z = 227 [M+H] + . Example 1: Synthesis of Compound 1 (8-[(3R,4R)-3-amino-4-methylpyrrolidin-1-yl]quinoxaline-5-carbonitrile) [ka]

[0200] tert-Butyl N-[(3R,4R)-1-(8-cyanoquinoxalin-5-yl)-4-methylpyrrolidin-3-yl]carbamate: To a solution of 8-bromoquinoxaline-5-carbonitrile (900 mg, 3.85 mmol) and tert-butyl N-[(3R,4R)-4-methylpyrrolidin-3-yl]carbamate (808 mg, 4.03 mmol) in N,N-dimethylformamide (7 mL) was added DIEA (1.58 g, 12.20 mmol) at room temperature. o The mixture was stirred at RT for 3 h. When the reaction was complete, it was quenched by adding water (30 mL). The resulting mixture was extracted with ethyl acetate (50 mL x 3). The organic phases were combined, washed with brine, and dried over Na2SO4. The solvent was removed under reduced pressure to afford tert-butyl N-[(3R,4R)-1-(8-cyanoquinoxalin-5-yl)-4-methylpyrrolidin-3-yl]carbamate as a yellow solid (1.19 g, 44%). MS: m / z = 354.1 [M+H] + .

[0201] 8-[(3R,4R)-3-amino-4-methylpyrrolidin-1-yl]quinoxaline-5-: To a solution of tert-butyl N-[(3R,4R)-1-(8-cyanoquinoxalin-5-yl)-4-methylpyrrolidin-3-yl]carbamate (720 mg, 2.04 mmol) in MeOH was added hydrochloric acid solution (12 N, 6 mL, 72 mmol) at room temperature. The resulting mixture was stirred at room temperature for 6 hours. Upon completion of the reaction, the reaction mixture was concentrated under reduced pressure, and the residue was purified by prep-HPLC under the following conditions: column, XBridge Prep C18 OBD column, 250 mm, 5 μm, 13 nm; mobile phase, acetonitrile in water (containing 0.05% NH3.HO), 29% to 42% gradient in 8 min; detector, UV 254 nm. 8-[(3R,4R)-3-amino-4-methylpyrrolidin-1-yl]quinoxaline-5-carbonitrile was obtained as a white solid (490 mg, 95%).

[0202] Compound 1: HPLC: 98.1% purity, RT=1.05min. MS:m / z=254.3[M+H] + . 1 H NMR (400 MHz, methanol-d 4、 ppm) δ 8.79 (d, J = 1.8 Hz, 1 H), 8.73 (d, J = 1.8 Hz, 1 H), 7.97-7.89 (m, 1 H), 6.72 (d, J = 8.8 Hz, 1 H), 4.21-4.12 (m, 1 H), 4.05-3.95 (m, 1 H), 3.93-3.85 (m, 1 H), 3.79-3.69 (m, 1 H), 3.63-3.52 (m, 1 H), 2.53-2.38 (m, 1 H), 1.16 (d, J = 7.0 Hz, 3 H).

[0203] The following compounds were synthesized in a similar manner:

[0204] Compound 56 8-(3-amino-4-methyl-pyrrolidin-1-yl)-quinoxaline-5-carbonitrile hydrochloride: from 8-bromoquinoxaline-5-carbonitrile and (4-methyl-pyrrolidin-3-yl)-carbamic acid tert-butyl ester. HPLC: 92% purity, RT=1.50 min. MS: m / z=254 [M+H] + . Example 2: Synthesis of Compound 2 (N-[(3R,4R)-1-(8-cyanoquinoxalin-5-yl)-4-methylpyrrolidin-3-yl]-2-(1-methylpiperidin-4-yl)acetamide) [ka]

[0205] N-[(3R,4R)-1-(8-cyanoquinoxalin-5-yl)-4-methylpyrrolidin-3-yl]-2-(1-methylpiperidin-4-yl)acetamide: To a solution of 8-[(3R,4R)-3-amino-4-methylpyrrolidin-1-yl]quinoxaline-5-carbonitrile (48 mg, 0.19 mmol) in N,N-dimethylformamide (10 mL) was added 2-(1-methylpiperidin-4-yl)acetic acid (59 mg, 0.38 mmol), DIEA (145 mg, 1.13 mmol), and HATU (143 mg, 0.38 mmol) at room temperature. The resulting solution was stirred at room temperature for 14 hours. Upon completion of the reaction, the solvent was removed under reduced pressure, and the residue was purified by prep-HPLC under the following conditions: column, XBridge Shield RP18 OBD column, 150 mm, 5 μm, 13 nm; mobile phase, acetonitrile in water (containing 10 mM NH₄HCO₃), 13% to 40% gradient in 8 min; detector, UV 254 nm. N-[(3R,4R)-1-(8-cyanoquinoxalin-5-yl)-4-methylpyrrolidin-3-yl]-2-(1-methylpiperidin-4-yl)acetamide was obtained as a yellow solid (18 mg, 25%).

[0206] Compound 2: HPLC: 92.4% purity, RT=3.85min. MS:m / z=393.4[M+H]+ . 1 H NMR (400 MHz, methanol-d 4、 ppm) δ 8.81 (d, J = 1.8 Hz, 1 H), 8.75 (d, J = 1.8 Hz, 1 H), 7.95 (d, J = 8.7 Hz, 1 H), 6.75 (d, J = 8.7 Hz, 1 H), 4.64-4.56 (m, 1 H), 4.31-4.21 (m, 1 H), 4.08-3.92 (m, 2 H), 3.74-3.64 (m, 1 H), 2.89-2.78 (m, 2 H), 2.69-2.54 (m, 1 H), 2.29-2.09 (m, 5 H), 2.06-1.94 (m, 2 H), 1.83-1.62 (m, 3 H), 1.37-1.20 (m, 2 H), 1.10 (d, J = 6.8 Hz, 3 H).

[0207] The following compounds were synthesized in a similar manner:

[0208] Compound 7 (N-[(3R,4R)-1-(8-cyanoquinoxalin-5-yl)-4-methylpyrrolidin-3-yl]-2-(morpholin-4-yl)acetamide): Purified from 8-[(3R,4R)-3-amino-4-methylpyrrolidin-1-yl]quinoxaline-5-carbonitrile and 2-(morpholin-4-yl)acetic acid by prep-HPLC under the following conditions: Column: XBridge Prep C18 OBD column, 150 mm, 5 μm, 13 nm; Mobile phase: acetonitrile in water (containing 10 mM NH4HCO3 and 0.1% NH3.HO), 5% to 75% gradient in 7 min; Detector: UV 254 nm (18 mg, 25%, yellow solid). HPLC: 91.7% purity, RT = 2.14 min. MS: m / z=381.3 [M+H] + . 1 H NMR (400 MHz, methanol-d 4、ppm) δ 8.83 (d, J = 1.8 Hz, 1 H), 8.76 (d, J = 1.8 Hz, 1 H), 7.97 (d, J = 8.7 Hz, 1 H), 6.78 (d, J = 8.7 Hz, 1 H), 4.64-4.54 (m, 1 H), 4.31-4.21 (m, 1 H), 4.12-3.96 (m, 2 H), 3.76-3.62 (m, 5 H), 3.09-3.04 (m, 2 H), 2.71-2.59 (m, 1 H), 2.57-2.45 (m, 4 H), 1.12 (d, J = 6.8 Hz, 3 H).

[0209] Compound 8 (N-[(3R,4R)-1-(8-cyanoquinoxalin-5-yl)-4-methylpyrrolidin-3-yl]-2-(1,4-dimethylpiperidin-4-yl)acetamide): Purified by prep-HPLC from 8-[(3R,4R)-3-amino-4-methylpyrrolidin-1-yl]quinoxaline-5-carbonitrile (43 mg, 0.17 mmol) and 2-(1,4-dimethylpiperidin-4-yl)acetic acid under the following conditions: Column: XBridge Prep C18 OBD column, 150 mm, 5 μm, 13 nm; Mobile phase: acetonitrile in water (containing 10 mM NH4HCO3), 22% to 35% gradient in 7 min; Detector: UV 254 nm (12 mg, 17%, yellow solid). HPLC: 94.5% purity, RT = 4.31 min. MS: m / z=407.3 [M+H] + . 1 H NMR (300 MHz, DMSO-d 6、 ppm) δ 8.91 (d, J = 1.8 Hz, 1 H), 8.80 (d, J = 1.8 Hz, 1 H), 8.01 (d, J = 8.7 Hz, 1 H), 7.89 (d, J = 8.3 Hz, 1 H), 6.71 (d, J = 8.8 Hz, 1 H), 4.52-4.39 (m, 1 H), 4.19-4.09 (m, 1 H), 3.97-3.90 (m, 1 H), 3.79-3.55 (m, 2 H), 2.47-1.93 (m, 10 H), 1.55-1.39 (m, 2 H), 1.31-1.25 (m, 2 H), 1.08-0.88 (m, 6 H).

[0210] Compound 15 ((2S)-N-[(3R,4R)-1-(8-cyanoquinoxalin-5-yl)-4-methylpyrrolidin-3-yl]-2-hydroxy-3-methylbutanamide): Purified from 8-[(3R,4R)-3-amino-4-methylpyrrolidin-1-yl]quinoxaline-5-carbonitrile and (2S)-2-hydroxy-3-methylbutanoic acid by prep-HPLC under the following conditions: Column: XBridge Shield Prep C18 OBD column, 150 mm, 5 μm, 13 nm; Mobile phase: Acetonitrile in water (containing 10 mmol / L NH4HCO3 and 0.1% NH3.HO), 30% to 35% gradient in 7 min; Detector: UV 254 nm (29 mg, 27%, yellow solid). HPLC: 95.0% purity, RT = 3.35 min. MS: m / z=354.3 [M+H] + . 1 H NMR (400 MHz, DMSO-d 6、 ppm) δ 8.93 (d, J = 1.9 Hz, 1 H), 8.81 (d, J = 1.8 Hz, 1 H), 8.03 (d, J = 8.7 Hz, 1 H), 7.79 (d, J = 8.5 Hz, 1 H), 6.73 (d, J = 8.8 Hz, 1 H), 5.09 (d, J = 6.1 Hz, 1 H), 4.55-4.45 (m, 1 H), 4.21-4.12 (m, 1 H), 4.05-3.74 (m, 2 H), 3.71-3.58 (m, 2 H), 2.58-2.49 (m, 1 H), 1.98-1.85 (m, 1 H), 0.98 (d, J = 6.8Hz, 3H), 0.86 (d, J = 6.8 Hz, 3 H), 0.73 (d, J = 6.8 Hz, 3 H).

[0211] Compound 16 ((2R)-N-[(3R,4R)-1-(8-cyanoquinoxalin-5-yl)-4-methylpyrrolidin-3-yl]-2-hydroxy-3-methylbutanamide): Purified from 8-[(3R,4R)-3-amino-4-methylpyrrolidin-1-yl]quinoxaline-5-carbonitrile and (2R)-2-hydroxy-3-methylbutanoic acid by prep-HPLC under the following conditions: column, XBridge Shield Prep C18 OBD column, 150 mm, 5 μm, 13 nm; mobile phase, acetonitrile in water (containing 10 mmol / L NH4HCO3 and 0.1% NH3.HO), 35% to 39% gradient in 7 min; detector, UV 254 nm (14 mg, 26%, yellow solid). HPLC: 96.1% purity, RT = 2.16 min. MS: m / z=354.4 [M+H] + . 1 H NMR (300 MHz, methanol-d 4、 ppm) δ 8.79 (d, J = 1.8 Hz, 1 H), 8.73 (d, J = 1.8 Hz, 1 H), 7.94 (d, J = 8.7 Hz, 1 H), 6.75 (d, J = 8.7 Hz, 1 H), 4.61-4.54 (m, 1 H), 4.30-4.18 (m, 1 H), 4.11-3.91 (m, 2 H), 3.86-3.78 (m, 1 H), 3.76-3.63 (m, 1 H), 2.66-2.57 (m, 1 H), 2.11-1.96 (m, 1 H), 1.10 (d, J = 6.8 Hz, 3 H), 0.96 (d, J = 6.9 Hz, 3 H), 0.86 (d, J = 6.8 Hz, 3 H). Example 3: Synthesis of compounds 3 and 4 (N-[(3R,4R)-1-(8-cyanoquinoxalin-5-yl)-4-methylpyrrolidin-3-yl]-2-[(3S)-1-methylpiperidin-3-yl]acetamide and N-[(3R,4R)-1-(8-cyanoquinoxalin-5-yl)-4-methylpyrrolidin-3-yl]-2-[(3R)-1-methylpiperidin-3-yl]acetamide) [ka]

[0212] N-[(3R,4R)-1-(8-cyanoquinoxalin-5-yl)-4-methylpyrrolidin-3-yl]-2-[(3S)-1-methylpiperidin-3-yl]acetamide and N-[(3R,4R)-1-(8-cyanoquinoxalin-5-yl)-4-methylpyrrolidin-3-yl]-2-[(3R)-1-methylpiperidin-3-yl]acetamide: 8-[(3R, To a solution of 4R)-3-amino-4-methylpyrrolidin-1-yl]quinoxaline-5-carbonitrile (48 mg, 0.19 mmol) in N,N-dimethylformamide (4 mL) was added 2-(1-methylpiperidin-3-yl)acetic acid (59 mg, 0.38 mmol), HATU (144 mg, 0.38 mmol), and DIEA (147 mg, 1.14 mmol) at room temperature. The resulting solution was stirred at room temperature for 14 hours. When the reaction was complete, it was quenched by adding water (15 mL). The resulting mixture was extracted with DCM (30 mL x 3). The organic phases were combined, washed with brine, and dried over Na2SO4. The solvent was removed under reduced pressure, and the residue was first purified by prep-HPLC under the following conditions: column, XBridge C18 OBD Prep column, 150 mm, 5 μm; mobile phase, acetonitrile in water (containing 10 mM NH4HCO3), 25% to 31% gradient in 8 min; detector, UV 254 nm. The two diastereomers were then separated on chiral prep-HPLC under the following conditions: column, CHIRALPAK IG, 2 x 15 cm, 3 μm; mobile phase, 100% MeOH (containing 0.1% DEA) in 20 min; detector, UV 254 nm.

[0213] Isomer 1: (18 mg, 25%, yellow solid) HPLC: 90.3% purity, RT=3.55min. MS:m / z=393.4[M+H] + . 1 H NMR (400 MHz, methanol-d 4、ppm) δ 8.82 (d, J = 1.7 Hz, 1 H), 8.76 (d, J = 1.8 Hz, 1 H), 7.96 (d, J = 8.7 Hz, 1 H), 6.77 (d, J = 8.7 Hz, 1 H), 4.62-4.57 (m, 1 H), 4.32-4.22 (m, 1 H), 4.09-3.94 (m, 2 H), 3.75-3.65 (m, 1 H), 2.92-2.78 (m, 2 H), 2.68-2.56 (m, 1 H), 2.27 (s, 3 H), 2.18-2.11 (m, 2 H), 2.04-1.99 (m, 2 H), 1.84-1.64 (m, 3 H), 1.64-1.53 ​​(m, 1 H), 1.11 (d, J = 6.8 Hz, 3 H), 1.03-0.90 (m, 1 H).

[0214] Isomer 2: (11 mg, 13%, yellow solid) HPLC: 95.3% purity, RT=5.07min. MS:m / z=393.1[M+H] + . 1 H NMR (300 MHz, methanol-d 4、 ppm) δ 8.83-8.69 (m, 2 H), 7.94 (d, J = 8.7 Hz, 1 H), 6.74 (d, J = 8.8 Hz, 1 H), 4.60-4.53 (m, 1 H), 4.30-4.18 (m, 1 H), 4.08-3.89 (m, 2 H), 3.74-3.60 (m, 1 H), 3.04-2.97 (m, 2 H), 2.67-2.51 (m, 1 H), 2.40 (s, 3 H), 2.32-1.96 (m, 6 H), 1.82-1.72 (m, 2 H), 1.67-1.56 (m, 1 H), 1.30-0.85 (m, 4 H).

[0215] The following compounds were synthesized in a similar manner:

[0216] Compounds 5 and 6 (N-[(3R,4R)-1-(8-cyanoquinoxalin-5-yl)-4-methylpyrrolidin-3-yl]-2-[(3S)-1-methylpyrrolidin-3-yl]acetamide and N-[(3R,4R)-1-(8-cyanoquinoxalin-5-yl)-4-methylpyrrolidin-3-yl]-2-[(3R)-1-methylpyrrolidin-3-yl]acetamide): were purified by prep-HPLC from 8-[(3R,4R)-3-amino-4-methylpyrrolidin-1-yl]quinoxaline-5-carbonitrile and 2-(1-methylpyrrolidin-3-yl)acetic acid under the following conditions: column, XBridge C18 OBD Prep Column, 150 mm, 5 μm; mobile phase, acetonitrile (10 mM) in water. NH4HCO3 and 0.1% NH3.H2O), 15% to 45% gradient in 8 min; detector, UV 254 nm. The two diastereomers were separated on chiral prep-HPLC under the following conditions: column, CHIRALPAK IC-3, 0.46 x 10 cm, 3 um; mobile phase, MtBE (containing 0.1% DEA) in 70% isocratic methanol in 30 min; detector, UV 254 nm. Isomer 1: (13 mg, 15%, yellow solid) HPLC: 97.3% purity, RT = 3.00 min. MS: m / z = 379.3 [M+H] + . 1 H NMR (300 MHz, DMSO-d 6、 ppm) δ 8.91 (d, J = 1.8 Hz, 1 H), 8.80 (d, J = 1.8 Hz, 1 H), 8.00 (d, J = 8.4 Hz, 2 H), 6.71 (d, J = 8.8 Hz, 1 H), 4.51-4.40 (m, 1 H), 4.15-4.08 (m, 1 H), 3.96-3.89 (m, 1 H), 3.80-3.73 (m, 1 H), 3.61-3.54 (m, 1 H), 2.37-2.26 (m, 5 H), 2.23-1.98 (m, 6 H), 1.91-1.75 (m, 1 H), 1.36-1.21 (m, 1 H), 0.94 (d, J = 6.8 Hz, 3 H). Isomer 2: (15 mg, 16%, yellow solid) HPLC: 94.1% purity, RT = 4.39 min. MS: m / z = 379.3 [M+H] + . 1 H NMR (300 MHz, メタノール-d 4、 ppm) δ 8.82-8.69 (m, 2 H), 7.93 (d, J = 8.7 Hz, 1 H), 6.73 (d, J = 8.7 Hz, 1 H), 4.59-4.52 (m, 1 H), 4.29-4.17 (m, 1 H), 4.08-3.88 (m, 2 H), 3.73-3.60 (m, 1 H), 2.80-2.68 (m, 1 H), 2.66-2.52 (m, 4 H), 2.33-2.17 (m, 6 H), 2.14-1.95 (m, 1 H), 1.57-1.39 (m, 1 H), 1.07 (d, J = 6.8 Hz, 3 H).

[0217] Compounds 13 and 14 ((2S)-N-[(3R,4R)-1-(8-cyanoquinoxalin-5-yl)-4-methylpyrrolidin-3-yl]-2-cyclopropyl-2-hydroxyacetamide and (2R)-N-[(3R,4R)-1-(8-cyanoquinoxalin-5-yl)-4-methylpyrrolidin-3-yl]-2-cyclopropyl-2-hydroxyacetamide) were purified by prep-HPLC from 8-[(3R,4R)-3-amino-4-methylpyrrolidin-1-yl]quinoxaline-5-carbonitrile (52 mg, 0.20 mmol) and 2-cyclopropyl-2-hydroxyacetic acid under the following conditions: column, XBridge Prep C18 OBD column, 150 mm, 5 μm; mobile phase, acetonitrile in water (10 mmol / L). NH4HCO3 and 0.1% NH3.H2O), 5% to 62% gradient in 7 min; detector, UV 254 nm. The two diastereomers were separated on chiral prep-HPLC under the following conditions: column, CHIRALPAK IA-3, 0.46 x 5 cm, 3 um; mobile phase, 50% isocratic hexane in EtOH (containing 0.1% DEA) in 15 min; detector, UV 254 nm. Isomer 1: (20 mg, 27%, yellow solid) HPLC: 96.1% purity, RT = 1.17 min. MS: m / z = 352.2 [M+H] + . 1 H NMR (400 MHz, DMSO-d 6、ppm) δ 8.93 (d, J = 1.8 Hz, 1 H), 8.82 (d, J = 1.8 Hz, 1 H), 8.04 (d, J = 8.7 Hz, 1 H), 7.78 (d, J = 8.5 Hz, 1 H), 6.74 (d, J = 8.8 Hz, 1 H), 5.09 (d, J = 6.1 Hz, 1 H), 4.52-4.47 (m, 1 H), 4.21-4.16 (m, 1 H), 4.00-3.95 (m, 1 H), 3.89-3.81 (m, 1 H), 3.62-3.57 (m, 1 H), 3.45 (t, J = 6.4 Hz, 1 H), 2.61-2.50 (m, 1 H), 1.08-0.95 (m, 4 H), 0.40-0.22 (m, 4 H). Isomer 2: (11 mg, 13%, yellow solid) HPLC: 97.4% purity, RT=1.19min. MS:m / z=352.2[M+H] + . 1 H NMR (400 MHz, DMSO-d 6、 ppm) δ 8.94 (d, J = 1.7 Hz, 1 H), 8.83 (d, J = 1.7 Hz, 1 H), 8.05 (d, J = 8.6 Hz, 1 H), 7.70 (d, J = 8.4 Hz, 1 H), 6.74 (d, J = 8.7 Hz, 1 H), 5.28 (d, J = 5.7 Hz, 1 H), 4.56-4.46 (m, 1 H), 4.25-4.15 (m, 1 H), 4.03-3.94 (m, 1 H), 3.91-3.83 (m, 1 H), 3.67-3.53 (m, 2 H), 2.61-2.52 (m, 1 H), 1.12-1.02 (m, 1 H), 1.00 (d, J = 6.8 Hz, 3H), 0.41-0.25 (m, 4H). Example 4: Synthesis of Compound 9 (N-[(3R,4R)-1-(8-cyanoquinoxalin-5-yl)-4-methylpyrrolidin-3-yl]-2-(4-fluoropiperidin-4-yl)acetamide) [ka]

[0218] tert-Butyl 4-([[(3R,4R)-1-(8-cyanoquinoxalin-5-yl)-4-methylpyrrolidin-3-yl]carbamoyl]methyl)-4-fluoropiperidine-1-carboxylate: To a solution of 8-[(3R,4R)-3-amino-4-methylpyrrolidin-1-yl]quinoxaline-5-carbonitrile (129 mg, 0.51 mmol) in DCM (8 mL) was added 2-[1-[(tert-butoxy)carbonyl]-4-fluoropiperidin-4-yl]acetic acid (266 mg, 1.02 mmol), HATU (388 mg, 1.02 mmol), and DIEA (394 mg, 3.06 mmol) at room temperature. The resulting solution was stirred at room temperature for 16 h. Upon completion of the reaction, the reaction mixture was concentrated under reduced pressure, and the residue was purified by prep-HPLC under the following conditions: column, XBridge Prep C18 OBD column, 150 mm, 5 μm, 13 nm; mobile phase, acetonitrile in water (containing 10 mM NH4HCO3 and 0.1% NH3.H2O), 40% to 53% gradient in 7 min; detector, UV 254 nm. 4-([[(3R,4R)-1-(8-cyanoquinoxalin-5-yl)-4-methylpyrrolidin-3-yl]carbamoyl]methyl)-4-fluoropiperidine-1-carboxylate was obtained as a yellow solid (76 mg, 30%). MS: m / z = 497.3 [M+H] + .

[0219] N-[(3R,4R)-1-(8-cyanoquinoxalin-5-yl)-4-methylpyrrolidin-3-yl]-2-(4-fluoropiperidin-4-yl)acetamide: To a solution of tert-butyl 4-([[(3R,4R)-1-(8-cyanoquinoxalin-5-yl)-4-methylpyrrolidin-3-yl]carbamoyl]methyl)-4-fluoropiperidine-1-carboxylate (76 mg, 0.15 mmol) in MeOH (4 mL) was added hydrochloric acid solution (12 N, 4 mL, 48 mmol) at room temperature. The resulting solution was stirred at room temperature for 12 h. Upon completion of the reaction, the reaction mixture was concentrated under reduced pressure, and the residue was purified by prep-HPLC under the following conditions: column, XBridge Prep C18 OBD column, 150 mm, 5 μm, 13 nm; mobile phase, acetonitrile in water (containing 10 mM NH4HCO3 and 0.1% NH3.HO), 15% to 42% gradient in 7 min; detector, UV 254 nm. N-[(3R,4R)-1-(8-cyanoquinoxalin-5-yl)-4-methylpyrrolidin-3-yl]-2-(4-fluoropiperidin-4-yl)acetamide was obtained as a yellow solid (13 mg, 19%).

[0220] Compound 9:MS:m / z=497.3[M+H] + . 1 H NMR (300 MHz, DMSO-d 6、 ppm) δ 8.95 (d, J = 1.8 Hz, 1 H), 8.84 (d, J = 1.8 Hz, 1 H), 8.13-8.01 (m, 2 H), 6.75 (d, J = 8.8 Hz, 1 H), 4.52-4.43 (m, 1 H), 4.26-3.86 (m, 2 H), 3.84-3.74 (m, 1 H), 3.62-3.52 (m, 1 H), 2.74-2.61 (m, 4 H), 2.50-2.36 (m, 3 H), 1.76-1.55 (m, 4 H), 1.00 (d, J = 6.8 Hz, 3 H). Example 5: Synthesis of Compound 10 (N-[(3R,4R)-1-(8-cyanoquinoxalin-5-yl)-4-methylpyrrolidin-3-yl]-2-(4-fluoro-1-methylpiperidin-4-yl)acetamide) [ka]

[0221] N-[(3R,4R)-1-(8-cyanoquinoxalin-5-yl)-4-methylpyrrolidin-3-yl]-2-(4-fluoro-1-methylpiperidin-4-yl)acetamide: N-[(3R,4R)-1-(8-cyanoquinoxalin-5-yl)-4-methylpyrrolidin-3-yl]-2-(4-fluoropiperidin-4-yl)acetamide (32 mg, 0.08 mmol) in MeOH (3 mL) (HCl). n (44 mg, 0.49 mmol), NaOAC (135 mg, 1.64 mmol), and NaBH (15 mg, 0.41 mmol) were added at room temperature. The resulting solution was stirred at room temperature for 3 h. Upon completion of the reaction, the reaction mixture was concentrated under reduced pressure, and the residue was purified by prep-HPLC under the following conditions: column, XBridge Shield RP18 OBD column, 150 mm, 5 μm, 13 nm; mobile phase, acetonitrile in water (containing 10 mM NHHCO), 35% to 65% gradient in 7 min; detector, UV 254 nm. N-[(3R,4R)-1-(8-cyanoquinoxalin-5-yl)-4-methylpyrrolidin-3-yl]-2-(4-fluoro-1-methylpiperidin-4-yl)acetamide was obtained as a yellow solid (7 mg, 21%).

[0222] Compound 10: HPLC: 90.3% purity, RT=5.76min. MS:m / z=411.3[M+H] + . 1 H NMR (300 MHz, methanol-d 4、ppm) δ 8.78 (d, J = 1.8 Hz, 1 H), 8.71 (d, J = 1.8 Hz, 1 H), 7.92 (d, J = 8.7 Hz, 1 H), 6.73 (d, J = 8.7 Hz, 1 H), 4.62-4.52 (m, 1 H), 4.31-4.16 (m, 1 H), 4.11-3.88 (m, 2 H), 3.73-3.59 (m, 1 H), 3.38-3.31 (m, 2 H), 3.22-3.06 (m, 2 H), 2.80 (s, 3 H), 2.73-2.52 (m, 3 H), 2.25-1.96 (m, 4 H), 1.09 (d, J = 6.8 Hz, 3H). Example 6: Synthesis of compounds 11 and 12 (N-[(3R,4R)-1-(8-cyanoquinoxalin-5-yl)-4-methylpyrrolidin-3-yl]-2-[(4S)-3,3-difluoro-1-methylpiperidin-4-yl]acetamide and N-[(3R,4R)-1-(8-cyanoquinoxalin-5-yl)-4-methylpyrrolidin-3-yl]-2-[(4R)-3,3-difluoro-1-methylpiperidin-4-yl]acetamide) [ka]

[0223] tert-Butyl 4-(2-ethoxy-2-oxoethylidene)-3,3-dimethylpiperidine-1-carboxylate: 0 o In C, sodium hydride (102 mg, 4.25 mmol) was added to a solution of ethyl (diethoxyphosphoryl) formate (950 mg, 4.52 mmol) in THF (50 mL). o C. The resulting mixture was stirred for 15 min, and then tert-butyl 3,3-difluoro-4-oxopiperidine-1-carboxylate (798 mg, 3.39 mmol) was added at 0. o The reaction mixture was added at 0 oThe mixture was stirred at RT for 0.5 h, warmed to room temperature, and stirred at room temperature for 3 h. When the reaction was complete, it was quenched by adding water (20 mL). The resulting mixture was extracted with ethyl acetate (40 mL x 3). The organic phases were combined, washed with brine, and dried over Na2SO4. The solvent was removed under reduced pressure, and the residue was purified by flash chromatography eluting with EtOAc in petroleum ether (0% to 50% gradient) to afford tert-butyl 4-(2-ethoxy-2-oxoethylidene)-3,3-dimethylpiperidine-1-carboxylate as a colorless oil (560 mg, 35%). MS: m / z = 205.9 [M -100 + 1] + .

[0224] tert-Butyl 4-(2-ethoxy-2-oxoethyl)-3,3-difluoropiperidine-1-carboxylate: To a solution of tert-butyl 4-(2-ethoxy-2-oxoethylidene)-3,3-difluoropiperidine-1-carboxylate (1.11 g, 3.62 mmol) in EtOH (30 mL) was added palladium on carbon (138 mg, 0.13 mmol) under a nitrogen atmosphere. The reaction vessel was evacuated and purged with hydrogen. The reaction mixture was then hydrogenated under a hydrogen atmosphere using a hydrogen balloon at room temperature for 3 h. Upon completion of the reaction, the reaction mixture was filtered through a Celite pad, and the filtrate was concentrated under reduced pressure to afford tert-butyl 4-(2-ethoxy-2-oxoethyl)-3,3-difluoropiperidine-1-carboxylate as a colorless oil (1.01 g, 92%). MS: m / z = 207.9 [M-100+1] + .

[0225] 2-[1-[(tert-butoxy)carbonyl]-3,3-difluoropiperidin-4-yl]acetic acid: To a solution of tert-butyl 4-(2-ethoxy-2-oxoethyl)-3,3-difluoropiperidine-1-carboxylate (1.01 g, 3.29 mmol) in tetrahydrofuran (25 mL) was added potassium hydroxide solution (500 mg, 12.5 mmol in 25 mL of water) at room temperature. The resulting mixture was cooled to 60°C.o The mixture was stirred at RT for 16 h at RT. After completion of the reaction, the reaction mixture was concentrated under vacuum. The pH value of the residual mixture was adjusted to 5 with hydrochloric acid solution (4N), and then the resulting mixture was extracted with ethyl acetate (50 mL x 3). The organic phases were combined, washed with brine, and dried over Na2SO4. The solvent was removed under reduced pressure to yield 2-[1-[(tert-butoxy)carbonyl]-3,3-difluoropiperidin-4-yl]acetic acid as a colorless oil (504 mg, 42%). MS: m / z = 280.1 [M+H] + .

[0226] 2-4-([[(3R,4R)-1-(8-cyanoquinoxalin-5-yl)-4-methylpyrrolidin-3-yl]carbamoyl]methyl)-3,3-difluoropiperidine-1-carboxylate: To a solution of 8-[(3R,4R)-3-amino-4-methylpyrrolidin-1-yl]quinoxaline-5-carbonitrile (208 mg, 0.82 mmol) in DCM (14 mL) was added 2-[1-[(tert-butoxy)carbonyl]-3,3-difluoropiperidin-4-yl]acetic acid (581 mg, 2.08 mmol), HATU (620 mg, 1.63 mmol), and DIEA (633 mg, 4.90 mmol) at room temperature. The resulting solution was stirred at room temperature for 1 h. When the reaction was complete, it was quenched by adding water (15 mL). The resulting mixture was extracted with ethyl acetate (50 mL x 3). The organic phases were combined, washed with brine, and dried over Na2SO4. The solvent was removed under reduced pressure to yield tert-butyl 4-([[(3R,4R)-1-(8-cyanoquinoxalin-5-yl)-4-methylpyrrolidin-3-yl]carbamoyl]methyl)-3,3-difluoropiperidine-1-carboxylate as a yellow solid (320 mg, crude). MS: m / z = 515.3 [M+H] + .

[0227] N-[(3R,4R)-1-(8-cyanoquinoxalin-5-yl)-4-methylpyrrolidin-3-yl]-2-[(4S)-3,3-difluoro-1-methylpiperidin-4-yl]acetamide and N-[(3R,4R)-1-(8-cyanoquinoxalin-5-yl)-4-methylpyrrolidin-3-yl]-2-[(4R)-3,3-difluoro-1-methylpiperidin-4-yl]acetamide: tert-butyl To a solution of 4-([[(3R,4R)-1-(8-cyanoquinoxalin-5-yl)-4-methylpyrrolidin-3-yl]carbamoyl]methyl)-3,3-difluoropiperidine-1-carboxylate (320 mg, crude) in HCOOH (26 mL) was added formalin (10%, 15 mL, 54 mmol) at room temperature. The resulting mixture was stirred for 140 minutes. o The mixture was stirred at RT for 1 h. After completion of the reaction, the reaction mixture was concentrated under reduced pressure, and the residue was purified by prep-HPLC under the following conditions: column, XBridge C18 OBD Prep column, 150 mm, 5 μm; mobile phase, acetonitrile in water (containing 10 mM NH4HCO3 and 0.1% NH3.H2O), 30% to 44% gradient in 8 min; detector, UV 254 nm. The two diastereomers were then separated by chiral prep-HPLC under the following conditions: column, CHIRALPAK IA-3, 0.46 x 5 cm, 3 μm; mobile phase, hexane in 50% isocratic ethanol (containing 0.1% DEA) in 15 min; detector, UV 254 nm.

[0228] Isomer 1: (93 mg, 59%, yellow solid) HPLC: 93.2% purity, RT=2.67min. MS:m / z=429.4[M+H] + . 1 H NMR (300 MHz, methanol-d 4、ppm) δ 8.78 (d, J = 1.8 Hz, 1 H), 8.72 (d, J = 1.8 Hz, 1 H), 7.93 (d, J = 8.7 Hz, 1 H), 6.73 (d, J = 8.7 Hz, 1 H), 4.62-4.53 (m, 1 H), 4.30-4.17 (m, 1 H), 4.07-3.88 (m, 2 H), 3.73-3.60 (m, 1H), 3.03-2.97 (m, 1 H), 2.86-2.75 (m, 1 H), 2.69-2.50 (m, 2 H), 2.34-2.01 (m, 7 H), 1.85-1.72 (m, 1 H),1.60-1.42 (m, 1 H), 1.07 (d, J = 6.9 Hz, 3 H).

[0229] Isomer 2: (11 mg, 13%, yellow solid) HPLC: 93.1% purity, RT=2.74min. MS:m / z=429.4[M+H] + . 1 H NMR (300 MHz, methanol-d 4、 ppm) δ 8.82-8.69 (m, 2 H), 7.93 (d, J = 8.7 Hz, 1 H), 6.73 (d, J = 8.7 Hz, 1 H), 4.60-4.53 (m, 1 H), 4.31-4.18 (m, 1 H), 4.07-3.89 (m, 2 H), 3.73-3.60 (m, 1 H), 3.04-2.94 (m, 1 H), 2.81-2.70 (m, 1 H), 2.69-2.50 (m, 2 H), 2.38-1.89 (m, 7 H), 1.79-1.68 (m, 1 H), 1.56-1.41 (m, 1 H), 1.07 (d, J = 6.9 Hz, 3 H). Example 7: Synthesis of Compound 17 ((3R,4R)-4-methyl-1-[8-(trifluoromethyl)quinolin-5-yl]pyrrolidin-3-amine) [ka]

[0230] tert-Butyl N-[(3R,4R)-4-methyl-1-[8-(trifluoromethyl)quinolin-5-yl]pyrrolidin-3-yl]carbamate: To a solution of 5-bromo-8-(trifluoromethyl)quinoline (245 mg, 0.89 mmol) in N,N-dimethylformamide (5 mL) was added tert-butyl N-[(3R,4R)-4-methylpyrrolidin-3-yl]carbamate (241 mg, 1.20 mmol), Pd(dba).CHCl (124 mg, 0.12 mmol), KPO (768 mg, 3.62 mmol), and DavePhos (93 mg, 0.23 mmol) at room temperature. The resulting mixture is 130 o The mixture was stirred at RT for 3 h. After completion of the reaction, the solids in the reaction mixture were filtered off, and the filtrate was diluted with water (20 mL). The resulting mixture was extracted with DCM (50 mL x 3). The organic phases were combined, washed with brine, and dried over Na2SO4. The solvent was removed under reduced pressure, and the residue was purified by flash chromatography eluting with EtOAc in hexane (0% to 20% gradient) to yield tert-butyl N-[(3R,4R)-4-methyl-1-[8-(trifluoromethyl)quinolin-5-yl]pyrrolidin-3-yl]carbamate as a brown solid (298 mg, 85%). MS: m / z = 396.2 [M+H] + .

[0231] (3R,4R)-4-Methyl-1-[8-(trifluoromethyl)quinolin-5-yl]pyrrolidin-3-amine: To a solution of tert-butyl N-[(3R,4R)-4-methyl-1-[8-(trifluoromethyl)quinolin-5-yl]pyrrolidin-3-yl]carbamate (50 mg, 0.13 mmol) in methanol (3 mL) was added hydrochloric acid solution (6 M in dioxane, 2 mL, 12 mmol) at room temperature. The resulting solution was stirred for 2 h at room temperature. Upon completion of the reaction, the reaction mixture was concentrated under reduced pressure, and the residue was purified by prep-HPLC under the following conditions: column, XBridge Shield Prep C18 OBD column, 150 mm, 5 μm, 13 nm; mobile phase, acetonitrile in water (containing 10 mmol / L NH4HCO3 and 0.1% NH3.HO), 30% to 51% gradient in 7 min; detector, UV 254 nm. (3R,4R)-4-methyl-1-[8-(trifluoromethyl)quinolin-5-yl]pyrrolidin-3-amine was obtained as a yellow solid (25 mg, 64%).

[0232] Compound 17: HPLC: 93.4% purity, RT=2.08min. MS:m / z=296.0[M+H] + . 1 H NMR (300 MHz, DMSO-d 6、 ppm) δ 8.89 (d, J = 3.9 Hz, 1 H), 8.70 (d, J = 8.8 Hz, 1 H), 7.86 (d, J = 8.4 Hz, 1 H), 7.51-7.40 (m, 1 H), 6.67 (d, J = 8.5 Hz, 1 H), 3.92-3.80 (m, 1 H), 3.58-3.43 (m, 3H), 3.38-3.27 (m, 1 H), 2.37-2.22 (m, 1 H), 1.03 (d, J = 6.8 Hz, 3 H). Example 8: Synthesis of Compound 18 (N-[(3R,4R)-4-methyl-1-[8-(trifluoromethyl)quinolin-5-yl]pyrrolidin-3-yl]-2-(1-methylpiperidin-4-yl)acetamide) [ka]

[0233] N-[(3R,4R)-4-Methyl-1-[8-(trifluoromethyl)quinolin-5-yl]pyrrolidin-3-yl]-2-(1-methylpiperidin-4-yl)acetamide: To a solution of (3R,4R)-4-methyl-1-[8-(trifluoromethyl)quinolin-5-yl]pyrrolidin-3-amine (38 mg, 0.13 mmol) in N,N-dimethylformamide (3 mL) was added 2-(1-methylpiperidin-4-yl)acetic acid (63 mg, 0.40 mmol), DIEA (31 mg, 0.24 mmol), and HATU (307 mg, 0.81 mmol, 6.36 equiv, 95%) at room temperature. The resulting solution was stirred at room temperature for 3 h. Upon completion of the reaction, the reaction mixture was concentrated under reduced pressure, and the residue was purified by prep-HPLC under the following conditions: column, XBridge Shield Prep C18 OBD column, 150 mm, 5 μm, 13 nm; mobile phase, acetonitrile in water (containing 10 mmol / L NH4HCO3 and 0.1% NH3.HO), 35% to 42% gradient in 7 min; detector, UV 254 nm. N-[(3R,4R)-4-methyl-1-[8-(trifluoromethyl)quinolin-5-yl]pyrrolidin-3-yl]-2-(1-methylpiperidin-4-yl)acetamide was obtained as a yellow solid (25 mg, 44%).

[0234] Compound 18: HPLC: 98.4% purity, RT=0.90min. MS: m / z = 435.2 [M+H] + . 1 H NMR (300 MHz, DMSO-d 6、ppm) δ 8.95-8.86 (m, 1 H), 8.69 (d, J = 8.8, 1.7 Hz, 1 H), 7.98-7.84 (m, 2 H), 7.54-7.43 (m, 1 H), 6.72 (d, J = 8.5 Hz, 1 H), 4.54-4.40 (m, 1 H), 4.04-3.92 (m, 1 H), 3.68-3.56 (m, 1 H), 3.58-3.45 (m, 1 H), 3.37-3.25 (m, 2 H), 2.74-2.60 (m, 2 H), 2.14-1.99 (m, 5 H), 1.78 (d, J = 9.4 Hz, 2 H), 1.61-1.44 (m, 3 H), 1.19-1.02 (m, 2 H), 0.97 (d, J = 6.8 Hz, 3 H).

[0235] The following compounds were synthesized in a similar manner:

[0236] Compound 23 (N-[(3R,4R)-4-methyl-1-[8-(trifluoromethyl)quinolin-5-yl]pyrrolidin-3-yl]-2-(morpholin-4-yl)acetamide): Purified from (3R,4R)-4-methyl-1-[8-(trifluoromethyl)quinolin-5-yl]pyrrolidin-3-amine and 2-(morpholin-4-yl)acetic acid on prep-HPLC under the following conditions: Column, XBridge Shield Prep C18 OBD column, 150 mm, 5 um, 13 nm; Mobile phase, acetonitrile in water (containing 0.05% NH3.HO), gradient 39% to 59% in 8 min; Detector, UV 254 nm (35 mg, 39%, yellow solid). HPLC: 96.3% purity, RT = 2.80 min. MS: m / z = 423.1 [M+H] + . 1 H NMR (300 MHz, DMSO-d 6、ppm) δ 8.96-8.87 (m, 1 H), 8.70 (d, J = 8.8, 1.7 Hz, 1 H), 7.95-7.80 (m, 2 H), 7.55-7.44 (m, 1 H), 6.76 (d, J = 8.5 Hz, 1 H), 4.52-4.43 (m, 1 H), 4.01-3.89 (m, 1 H), 3.72-3.37 (m, 7 H), 3.06-2.88 (m, 2 H), 2.59-2.51 (m, 1 H), 2.47-2.37 (m, 4 H), 0.98 (d, J = 6.8 Hz, 3 H).

[0237] Compound 24 (2-(1,4-dimethylpiperidin-4-yl)-N-[(3R,4R)-4-methyl-1-[8-(trifluoromethyl)quinolin-5-yl]pyrrolidin-3-yl]acetamide): Purified from (3R,4R)-4-methyl-1-[8-(trifluoromethyl)quinolin-5-yl]pyrrolidin-3-amine (60 mg, 0.20 mmol) and 2-(1,4-dimethylpiperidin-4-yl)acetic acid on a prep-HPLC system using the following conditions: column, XBridge Shield Prep C18 OBD column, 150 mm, 5 μm, 13 nm; mobile phase, acetonitrile in water (containing 10 mmol / L NH4HCO3 and 0.1% NH3.HO), 40% to 52% gradient in 10 min; detector, UV 254 nm (45 mg, 49%, yellow solid). HPLC: 98.4% purity, RT=4.24min. MS:m / z=449.4[M+H] + . 1 H NMR (300 MHz, DMSO-d 6、ppm) δ 8.95-8.87 (m, 1 H), 8.70 (d, J = 8.8, 1.7 Hz, 1 H), 7.94-7.84 (m, 2 H), 7.55-7.44 (m, 1 H), 6.72 (d, J = 8.5 Hz, 1 H), 4.50-4.41 (m, 1 H), 4.04-3.92 (m, 1 H), 3.66-3.46 (m, 2 H), 3.40-3.30 (m, 1 H), 2.36-2.30 (m, 2 H), 2.22-1.99 (m, 8 H), 1.58-1.44 (m, 2 H), 1.34-1.27 (m, 2 H), 1.03-0.91 (m, 6 H).

[0238] Compound 31 ((2S)-2-hydroxy-3-methyl-N-[(3R,4R)-4-methyl-1-[8-(trifluoromethyl)quinolin-5-yl]pyrrolidin-3-yl]butanamide): Purified from (3R,4R)-4-methyl-1-[8-(trifluoromethyl)quinolin-5-yl]pyrrolidin-3-amine and (2S)-2-hydroxy-3-methylbutanoic acid by prep-HPLC under the following conditions: Column: XBridge Shield Prep C18 OBD column, 150 mm, 5 μm; Mobile phase: acetonitrile in water (containing 10 mmol / L NH4HCO3 and 0.1% NH3.HO), 40% to 56% gradient in 7 min; Detector: UV 254 nm (42 mg, 53%, yellow solid). HPLC: 98.2% purity, RT = 4.89 min. MS: m / z=396.1 [M+H] + . 1 H NMR (300 MHz, DMSO-d 6、ppm) δ 8.94-8.86 (m, 1 H), 8.72 (d, J = 8.8, 1.7 Hz, 1 H), 7.88 (d, J = 8.4 Hz, 1 H), 7.67 (d, J = 8.5 Hz, 1 H), 7.53-7.42 (m, 1 H), 6.74 (d, J = 8.5 Hz, 1 H), 4.58-4.36 (m, 1 H), 3.96-3.84 (m, 1 H), 3.72-3.60 (m, 2 H), 3.59-3.42 (m, 2 H), 2.60-2.49 (m, 1 H), 2.06-1.91 (m, 1 H), 0.97 (d, J = 6.8 Hz, 3 H), 0.88 (d, J = 6.9 Hz, 3 H), 0.79 (d, J = 6.7 Hz, 3 H).

[0239] Compound 32 ((2R)-2-hydroxy-3-methyl-N-[(3R,4R)-4-methyl-1-[8-(trifluoromethyl)quinolin-5-yl]pyrrolidin-3-yl]butanamide): Purified from (3R,4R)-4-methyl-1-[8-(trifluoromethyl)quinolin-5-yl]pyrrolidin-3-amine (60 mg, 0.20 mmol) and (2S)-2-hydroxy-3-methylbutanoic acid by prep-HPLC under the following conditions: column, XBridge Shield Prep C18 OBD column, 150 mm, 5 μm; mobile phase, acetonitrile in water (containing 10 mmol / L NH4HCO3 and 0.1% NH3.HO), 40% to 56% gradient in 7 min; detector, UV 254 nm (42 mg, 52%, yellow solid). HPLC: 97.6% purity, RT=1.68min. MS:m / z=396.2[M+H] + . 1 H NMR (300 MHz, methanol-d 4、ppm) δ 8.89-8.80 (m, 1 H), 8.73 (d, J = 8.8, 1.7 Hz, 1 H), 7.91 (d, J = 8.5 Hz, 1 H), 7.53-7.42 (m, 1 H), 6.87 (d, J = 8.5 Hz, 1 H), 4.64-4.55 (m, 1 H), 4.03-3.86 (m, 2 H), 3.77-3.64 (m, 1 H), 3.61-3.43 (m, 2 H), 2.75-2.59 (m, 1 H), 2.14-1.97 (m, 1 H), 1.11 (d, J = 6.9 Hz, 3 H), 0.98 (d, J = 6.9 Hz, 3H), 0.80 (d, J = 6.8 Hz, 3 H). Example 9: Synthesis of compounds 19 and 20 (N-[(3R,4R)-4-methyl-1-[8-(trifluoromethyl)quinolin-5-yl]pyrrolidin-3-yl]-2-[(3S)-1-methylpiperidin-3-yl]acetamide and N-[(3R,4R)-4-methyl-1-[8-(trifluoromethyl)quinolin-5-yl]pyrrolidin-3-yl]-2-[(3R)-1-methylpiperidin-3-yl]acetamide) [ka]

[0240] N-[(3R,4R)-4-methyl-1-[8-(trifluoromethyl)quinolin-5-yl]pyrrolidin-3-yl]-2-[(3S)-1-methylpiperidin-3-yl]acetamide and N-[(3R,4R)-4-methyl-1-[8-(trifluoromethyl)quinolin-5-yl]pyrrolidin-3-yl]-2-[(3R)-1-methylpiperidin-3-yl]acetamide: (3R To a solution of (4R)-4-methyl-1-[8-(trifluoromethyl)quinolin-5-yl]pyrrolidin-3-amine (119 mg, 0.40 mmol) in N,N-dimethylformamide (5 mL), 2-(1-methylpiperidin-3-yl)acetic acid (152 mg, 0.97 mmol), DIEA (76 mg, 0.59 mmol), and HATU (735 mg, 1.93 mmol) were added at room temperature. The resulting mixture was stirred at room temperature for 3 h. Upon completion of the reaction, the reaction mixture was concentrated under reduced pressure, and the residue was first purified by prep-HPLC under the following conditions: column, XBridge Prep C18 OBD column, 150 mm, 5 μm; mobile phase, acetonitrile in water (containing 10 mmol / L NH4HCO3 and 0.1% NH3.HO), 40% to 70% gradient in 7 min; detector, UV 254 nm. The two diastereomers were then separated on chiral prep-HPLC under the following conditions: column, CHIRALPAK IG, 0.46x15cm, 3um; mobile phase, 88% isocratic hexane in EtOH (containing 0.1% DEA) in 30min; detector, UV 254nm.

[0241] Isomer 1: (20mg, 11%, yellow solid) HPLC: 97.8% purity, RT=2.39min. MS:m / z=435.3[M+H] + . 1 H NMR (300 MHz, DMSO-d 6、ppm) δ 8.95 - 8.87 (m, 1 H), 8.70 (d, J = 8.8, 1.7 Hz, 1 H), 8.00-7.85 (m, 2 H), 7.55-7.44 (m, 1 H), 6.73 (d, J = 8.5 Hz, 1 H), 4.58-4.37 (m, 1 H), 4.04-3.92 (m, 1 H), 3.69-3.57 (m, 1 H), 3.59-3.46 (m, 1 H), 3.38-3.27 (m, 1 H), 2.62-2.51 (m, 3 H), 2.14-1.94 (m, 5 H), 1.88-1.73 (m, 2 H), 1.65-1.48 (m, 3 H), 1.44-1.33 (m, 1 H), 0.97 (d, J = 6.7 Hz, 3 H), 0.91-0.74 (m, 1 H).

[0242] Isomer 2: (25 mg, 14%, yellow solid) HPLC: 99.5% purity, RT = 1.25 min. MS: m / z = 435.3 [M+H] + . 1 H NMR (300 MHz, DMSO-d 6、 ppm) δ 8.94-8.86 (m, 1 H), 8.69 (d, 1 H), 7.99-7.83 (m, 2 H), 7.54-7.43 (m, 1 H), 6.72 (d, J = 8.6 Hz, 1 H), 4.50-4.41 (m, 1 H), 4.04-3.92 (m, 1 H), 3.67-3.55 (m, 1 H), 3.58-3.45 (m, 1 H), 3.37-3.25 (m, 2 H), 2.62-2.52 (m, 2 H), 2.12-1.97 (m, 5 H), 1.92-1.73 (m, 2 H), 1.60-1.49 (m, 3 H), 1.45-1.34 (m, 1 H), 0.96 (d, J = 6.8 Hz, 3 H), 0.88-0.78 (m, 1 H).

[0243] The following compounds can be synthesized in similar ways:

[0244] Compounds 21 and 22 (N-[(3R,4R)-4-methyl-1-[8-(trifluoromethyl)quinolin-5-yl]pyrrolidin-3-yl]-2-[(3S)-1-methylpyrrolidin-3-yl]acetamide and N-[(3R,4R)-4-methyl-1-[8-(trifluoromethyl)quinolin-5-yl]pyrrolidin-3-yl]-2-[(3R)-1-methylpyrrolidin-3-yl]acetamide): Purified from (3R,4R)-4-methyl-1-[8-(trifluoromethyl)quinolin-5-yl]pyrrolidin-3-amine (109 mg, 0.37 mmol) and 2-(1-methylpyrrolidin-3-yl)acetic acid on prep-HPLC under the following conditions: column, XBridge Prep C18 OBD column, 150 mm, 5 μm; mobile phase, acetonitrile in water (containing 0.05% NH3.H2O), 39% to 54% gradient in 8 min; detector, UV 254 nm. The two diastereomers were separated on chiral prep-HPLC under the following conditions: column, CHIRALPAK IC-3, 0.46 x 15 cm, 3 μm; mobile phase, 50% isocratic hexane in IPA (containing 0.1% DEA) in 30 min; detector, UV 254 nm. Isomer 1: (20 mg, 11%, yellow solid) HPLC: 99.5% purity, RT = 2.87 min. MS: m / z = 421.2 [M+H] + . 1 H NMR (300 MHz, DMSO-d 6、ppm) δ 8.95 - 8.86 (m, 1 H), 8.69 (d, J = 8.7, 1.7 Hz, 1 H), 7.98 (d, J = 8.3 Hz, 1 H), 7.88 (d, J = 8.5 Hz, 1 H), 7.54 - 7.43 (m, 1 H), 6.72 (d, J = 8.5 Hz, 1 H), 4.50 - 4.41 (m, 1 H), 4.03 - 3.91 (m, 1 H), 3.67 - 3.45 (m, 2 H), 3.37 - 3.27 (m, 1 H), 2.58 - 2.30 (m, 5 H), 2.26 - 2.01 (m, 6 H), 1.91 - 1.73 (m, 1 H), 1.39 - 1.21 (m, 1 H), 0.96 (d, J = 6.7 Hz, 3 H). Isomer 2: (20 mg, 11%, yellow solid) HPLC: 98.8% purity, RT = 2.88 min. MS: m / z = 421.1 [M + H] + . 1 H NMR (300 MHz, DMSO-d 6、 ppm) δ 8.94 - 8.86 (m, 1 H), 8.74 - 8.64 (m, 1 H), 7.99 (d, J = 8.3 Hz, 1 H), 7.88 (d, J = 8.5 Hz, 1 H), 7.54 - 7.43 (m, 1 H), 6.72 (d, J = 8.5 Hz, 1 H), 4.49 - 4.42 (m, 1 H), 4.04 - 3.92 (m, 1 H), 3.67 - 3.45 (m, 2 H), 3.36 - 3.29 (m, 1 H), 2.59 - 2.33 (m, 5 H), 2.28 - 2.05 (m, 6 H), 1.97 - 1.79 (m, 1 H), 1.44 - 1.26 (m, 1 H), 0.96 (d, J = 6.8 Hz, 3 H).

[0245] Compounds 29 and 30 ((2S)-2-cyclopropyl-2-hydroxy-N-[(3R,4R)-4-methyl-1-[8-(trifluoromethyl)quinolin-5-yl]pyrrolidin-3-yl]acetamide and (2R)-2-cyclopropyl-2-hydroxy-N-[(3R,4R)-4-methyl-1-[8-(trifluoromethyl)quinolin-5-yl]pyrrolidin-3-yl]acetamide) were purified from (3R,4R)-4-methyl-1-[8-(trifluoromethyl)quinolin-5-yl]pyrrolidin-3-amine and 2-cyclopropyl-2-hydroxyacetic acid on prep-HPLC under the following conditions: column, XBridge Prep C18 OBD column, 150 mm, 5 μm; mobile phase, acetonitrile in water (10 mmol / L). NH4HCO3 and 0.1% NH3.H2O), gradient 35% to 52% in 7 min; detector, UV 254 nm. The two diastereomers were separated on chiral prep-HPLC under the following conditions: column, CHIRALPAK IA-3, 0.46 x 15 cm, 3 um; mobile phase, hexane in 90% isocratic EtOH (containing 0.1% DEA) in 30 min; detector, UV 254 nm. Isomer 1: (25 mg, 19%, yellow solid) HPLC: 99.8% purity, RT = 1.59 min. MS: m / z = 394.1 [M+H] + . 1 H NMR (300 MHz, DMSO-d 6、 ppm) δ 8.94-8.86 (m, 1 H), 8.71 (d, J = 8.7, 1.7 Hz, 1 H), 7.89 (d, J = 8.5 Hz, 1 H), 7.74 (d, J = 8.4 Hz, 1 H), 7.54-7.43 (m, 1 H), 6.73 (d, J = 8.5 Hz, 1 H), 4.53-4.46 (m, 1 H), 4.02-3.89 (m, 1 H), 3.72-3.60 (m, 1 H), 3.55-3.40 (m, 3 H), 2.60-2.51 (m, 1 H), 1.09-0.93 (m, 4 H), 0.43-0.21 (m, 4 H). Isomer 2: (18 mg, 10%, yellow solid) HPLC: 99.7% purity, RT=2.30min. MS:m / z=394.1[M+H] +. 1 H NMR (300 MHz, methanol-d 4、 ppm) δ 8.89-8.80 (m, 1 H), 8.75 (d, J = 8.8, 1.7 Hz, 1 H), 7.91 (d, J = 8.4 Hz, 1 H), 7.53-7.42 (m, 1 H), 6.88 (d, J = 8.4 Hz, 1 H), 4.66-4.59 (m, 1 H), 4.03-3.91 (m, 1 H), 3.76-3.63 (m, 2 H), 3.60-3.43 (m, 2 H), 2.75-2.59 (m, 1 H), 1.24-1.07 (m, 4 H), 0.57-0.37 (m, 4 H). Example 10: Synthesis of Compound 25 (2-(4-fluoropiperidin-4-yl)-N-[(3R,4R)-4-methyl-1-[8-(trifluoromethyl)quinolin-5-yl]pyrrolidin-3-yl]acetamide) [ka]

[0246] tert-Butyl 4-fluoro-4-([[(3R,4R)-4-methyl-1-[8-(trifluoromethyl)quinolin-5-yl]pyrrolidin-3-yl]carbamoyl]methyl)piperidine-1-carboxylate: To a solution of (3R,4R)-4-methyl-1-[8-(trifluoromethyl)quinolin-5-yl]pyrrolidin-3-amine (48 mg, 0.16 mmol) in N,N-dimethylformamide (1 mL) was added 2-[1-[(tert-butoxy)carbonyl]-4-fluoropiperidin-4-yl]acetic acid (82 mg, 0.31 mmol), DIEA (31 mg, 0.24 mmol), and HATU (307 mg, 0.81 mmol) at room temperature. The resulting solution was stirred at room temperature for 3 h. Upon completion of the reaction, the reaction mixture was diluted with water (10 mL), and the resulting mixture was extracted with ethyl acetate (30 mL x 3). The organic phases were combined, washed with brine, and dried over Na2SO4. The solvent was removed under reduced pressure to yield tert-butyl 4-fluoro-4-([[(3R,4R)-4-methyl-1-[8-(trifluoromethyl)quinolin-5-yl]pyrrolidin-3-yl]carbamoyl]methyl)piperidine-1-carboxylate as a yellow solid (80 mg, crude).

[0247] 2-(4-Fluoropiperidin-4-yl)-N-[(3R,4R)-4-methyl-1-[8-(trifluoromethyl)quinolin-5-yl]pyrrolidin-3-yl]acetamide: To a solution of tert-butyl 4-fluoro-4-([[(3R,4R)-4-methyl-1-[8-(trifluoromethyl)quinolin-5-yl]pyrrolidin-3-yl]carbamoyl]methyl)piperidine-1-carboxylate (80 mg, crude product) in methanol (3 mL) was added hydrochloric acid solution (6N in dioxane, 2 mL, 12 mmol) at room temperature. The resulting mixture was stirred at room temperature for 2 h. Upon completion of the reaction, the reaction mixture was concentrated under reduced pressure, and the residue was purified by prep-HPLC under the following conditions: column, XBridge Shield Prep C18 OBD column, 150 mm, 5 μm, 13 nm; mobile phase, acetonitrile in water (containing 10 mmol / L NH4HCO3 and 0.1% NH3.HO), gradient 32% to 38% in 7 min; detector, UV 254 nm. 2-(4-Fluoropiperidin-4-yl)-N-[(3R,4R)-4-methyl-1-[8-(trifluoromethyl)quinolin-5-yl]pyrrolidin-3-yl]acetamide was obtained as a yellow solid (12 mg, 1.3% stepwise).

[0248] Compound 25: HPLC: 95.2% purity, RT=2.93min. MS:m / z=439.4[M+H] + . 1 H NMR (300 MHz, DMSO-d 6、 ppm) δ 8.86-8.78 (m, 1 H), 8.61 (d, J = 8.8 Hz, 1 H), 7.80 (d, J = 8.5 Hz, 1 H), 7.67 (d, J = 7.5 Hz, 1 H), 7.45-7.34 (m, 1 H), 6.70 (d, J = 8.4 Hz, 1 H), 4.52-4.43 (m, 1 H), 4.10-3.90 (m, 1 H), 3.63-3.45 (m, 3 H), 2.81-2.60 (m, 4 H), 2.40-2.20 (m, 4 H), 1.85-1.55 (m, 4 H), 1.05-0.85 (m, 3 H). Example 11: Synthesis of Compound 26 (2-(4-fluoro-1-methylpiperidin-4-yl)-N-[(3R,4R)-4-methyl-1-[8-(trifluoromethyl)quinolin-5-yl]pyrrolidin-3-yl]acetamide) [ka]

[0249] 2-(4-Fluoro-1-methylpiperidin-4-yl)-N-[(3R,4R)-4-methyl-1-[8-(trifluoromethyl)quinolin-5-yl]pyrrolidin-3-yl]acetamide: 2-(4-Fluoro-1-methylpiperidin-4-yl)-N-[(3R,4R)-4-methyl-1-[8-(trifluoromethyl)quinolin-5-yl]pyrrolidin-3-yl]acetamide (48 mg, 0.11 mmol) in methanol (3 mL) (CHO) n (95 mg, 1.05 mmol), NaOAC (190 mg, 2.32 mmol), and NaBH (66 mg, 1.76 mmol) were added at room temperature. The resulting mixture was stirred at room temperature for 12 h. When the reaction was complete, it was quenched by adding water (5 mL). The resulting mixture was extracted with ethyl acetate (30 mL x 3). The organic phases were combined, washed with brine, and dried over NaSO. The solvent was removed under reduced pressure, and the residue was purified by prep-HPLC under the following conditions: column, XBridge Shield Prep C18 OBD column, 150 mm, 5 μm, 13 nm; mobile phase, acetonitrile in water (containing 10 mmol / L NHHCO and 0.1% NH.H0), 30% to 55% gradient in 7 min; detector, UV 254 nm. 2-(4-Fluoro-1-methylpiperidin-4-yl)-N-[(3R,4R)-4-methyl-1-[8-(trifluoromethyl)quinolin-5-yl]pyrrolidin-3-yl]acetamide was obtained as a yellow solid (20 mg, 40%).

[0250] Compound 26: HPLC: 98.3% purity, RT=3.39min. MS:m / z=453.5[M+H] + .1 H NMR (300 MHz, methanol-d 4、 ppm) δ 8.88-8.79 (m, 1 H), 8.74 (d, J = 8.8, 1.7 Hz, 1 H), 7.90 (d, J = 8.5 Hz, 1 H), 7.51-7.40 (m, 1 H), 6.83 (d, J = 8.5 Hz, 1 H), 4.65-4.54 (m, 1 H), 4.05-3.93 (m, 1 H), 3.72-3.60 (m, 1H), 3.62-3.49 (m, 1 H), 3.47-3.36 (m, 1 H), 2.74-2.45 (m, 5 H), 2.40-2.26 (m, 5 H), 1.95-1.89 (m, 3 H), 1.98-1.75 (m, 1 H), 1.09 (d, J = 6.8 Hz, 3 H). Example 12: Synthesis of compounds 27 and 28 (2-[(4S)-3,3-difluoro-1-methylpiperidin-4-yl]-N-[(3R,4R)-4-methyl-1-[8-(trifluoromethyl)quinolin-5-yl]pyrrolidin-3-yl]acetamide and 2-[(4R)-3,3-difluoro-1-methylpiperidin-4-yl]-N-[(3R,4R)-4-methyl-1-[8-(trifluoromethyl)quinolin-5-yl]pyrrolidin-3-yl]acetamide) [ka]

[0251] tert-Butyl 3,3-difluoro-4-([[(3R,4R)-4-methyl-1-[8-(trifluoromethyl)quinolin-5-yl]pyrrolidin-3-yl]carbamoyl]methyl)piperidine-1-carboxylate: To a solution of (3R,4R)-4-methyl-1-[8-(trifluoromethyl)quinolin-5-yl]pyrrolidin-3-amine (50 mg, 0.17 mmol) in dichloromethane (3 mL) was added 2-[1-[(tert-butoxy)carbonyl]-3,3-difluoropiperidin-4-yl]acetic acid (62 mg, 0.22 mmol), DIEA (32 mg, 0.24 mmol), and HATU (307 mg, 0.81 mmol) at room temperature. The resulting solution was stirred at room temperature for 3 h. When the reaction was complete, it was quenched by adding water (10 mL). The resulting mixture was extracted with ethyl acetate (50 mL x 3). The organic phases were combined, washed with brine, and dried over Na2SO4. The solvent was concentrated under reduced pressure to yield 3,3-difluoro-4-([[(3R,4R)-4-methyl-1-[8-(trifluoromethyl)quinolin-5-yl]pyrrolidin-3-yl]carbamoyl]methyl)piperidine-1-carboxylate as a yellow solid (80 mg, crude). MS: m / z = 557.5 [M+H] + .

[0252] tert-Butyl 3,3-difluoro-4-([[(3R,4R)-4-methyl-1-[8-(trifluoromethyl)quinolin-5-yl]pyrrolidin-3-yl]carbamoyl]methyl)piperidine-1-carboxylate: To a solution of tert-butyl 3,3-difluoro-4-([[(3R,4R)-4-methyl-1-[8-(trifluoromethyl)quinolin-5-yl]pyrrolidin-3-yl]carbamoyl]methyl)piperidine-1-carboxylate (80 mg, crude product) in methanol (3 mL) was added hydrochloric acid solution (6N in dioxane, 2 mL, 12 mmol) at room temperature. The resulting mixture was stirred at room temperature for 3 h. When the reaction was complete, it was quenched by adding water (10 mL). The pH value of the resulting mixture was adjusted to 8 using saturated NaHCO3 solution. The resulting mixture was extracted with ethyl acetate (50 mL x 3). The organic phases were combined, washed with brine, and dried over Na2SO4. The solvent was removed under reduced pressure to yield 2-(3,3-difluoropiperidin-4-yl)-N-[(3R,4R)-4-methyl-1-[8-(trifluoromethyl)quinolin-5-yl]pyrrolidin-3-yl]acetamide as a yellow solid (60 mg, crude). MS: m / z = 557.5 [M+H] + .

[0253] 2-[(4S)-3,3-Difluoro-1-methylpiperidin-4-yl]-N-[(3R,4R)-4-methyl-1-[8-(trifluoromethyl)quinolin-5-yl]pyrrolidin-3-yl]acetamide and 2-[(4R)-3,3-Difluoro-1-methylpiperidin-4-yl]-N-[(3R,4R)-4-methyl-1-[8-(trifluoromethyl)quinolin-5-yl]pyrrolidin-3-yl]acetamide: To a solution of 2-(3,3-difluoropiperidin-4-yl)-N-[(3R,4R)-4-methyl-1-[8-(trifluoromethyl)quinolin-5-yl]pyrrolidin-3-yl]acetamide (60 mg, crude) in methanol (4 mL) (CHO) n(110 mg, 2.45 mmol), NaOAc (200 mg, 4.86 mmol), and NaBH (72 mg, 3.79 mmol) were added at room temperature. The resulting mixture was stirred at room temperature for 16 h. Upon completion of the reaction, the reaction mixture was concentrated under reduced pressure, and the residue was first purified by prep-HPLC under the following conditions: column, XBridge Prep C18 OBD column, 150 mm, 5 μm; mobile phase, acetonitrile in water (containing 10 mmol / L NH4HCO3 and 0.1% NH3.H2O), 35% to 50% gradient in 8 min; detector, UV 254 nm. The two diastereomers were then separated on chiral prep-HPLC under the following conditions: column, CHIRALPAK IC-3, 0.46x15cm, 3um; mobile phase, 50% isocratic hexane in IPA (containing 0.1% DEA) in 20min; detector, UV 254nm.

[0254] Isomer 1: (12 mg, 15% over 3 steps, off-white solid) HPLC: 99.0% purity, RT = 3.06 min. MS: m / z = 471.5 [M+H] + . 1 H NMR (300 MHz, DMSO-d 6、 ppm) δ 8.96-8.87 (m, 1 H), 8.75-8.65 (m, 1 H), 8.12 (d, J = 8.4 Hz, 1 H), 7.90 (d, J = 8.4 Hz, 1 H), 7.55-7.44 (m, 1 H), 6.73 (d, J = 8.5 Hz, 1 H), 4.51-4.42 (m, 1 H), 4.06-3.94 (m, 1 H), 3.68-3.46 (m, 2 H), 2.98-2.86 (m, 1 H), 2.67-2.56 (m, 1 H), 2.50-2.44 (m, 2 H), 2.29-2.02 (m, 7 H), 1.98-1.84 (m, 1 H), 1.64-1.52 (m, 1 H), 1.38-1.20 (m, 1 H), 0.98 (d, J = 6.7 Hz, 3 H).

[0255] Isomer 2: (12 mg, 15% over 3 steps, yellow solid) HPLC: 99.5% purity, RT = 3.01 min. MS: m / z = 471.5 [M+H] + . 1 H NMR (300 MHz, DMSO-d 6、 ppm) δ 9.01-8.93 (m, 1 H), 8.83-8.71 (m, 1 H), 8.20 (d, J = 8.4 Hz, 1 H), 7.95 (d, J = 8.5 Hz, 1 H), 7.61-7.50 (m, 1 H), 6.79 (d, J = 8.6 Hz, 1 H), 4.64-4.40 (m, 1 H), 4.10 - 3.98 (m, 1 H), 3.74-3.62 (m, 1 H), 3.65-3.52 (m, 1 H), 3.14-2.83 (m, 2 H), 2.82-2.71 (m, 1 H), 2.53-2.47 (m, 1 H), 2.38-1.88 (m, 8 H), 1.81-1.70 (m, 1 H), 1.52-1.34 (m, 1 H), 1.02 (d, J = 6.7 Hz, 3 H). Example 13: Synthesis of Compound 33 ((3R,4R)-4-methyl-1-(8-methylquinolin-5-yl)pyrrolidin-3-amine) [ka]

[0256] 5-Bromo-8-methylquinoline: To a mixture of 5-bromo-2-methylaniline (980 mg, 5.27 mmol) in HCl solution (2 M, 30 mL) was added 3,3-diethoxyprop-1-ene (1.67 g, 12.80 mmol) at room temperature. The resulting mixture was stirred at 110 oThe mixture was stirred at RT for 16 h at RT. After completion of the reaction, the pH of the reaction mixture was adjusted to 7-8 with sodium bicarbonate solution (4 M). The resulting mixture was extracted with DCM (100 mL x 3). The organic phases were combined, washed with brine, and dried over Na2SO4. The organic phases were combined, washed with brine, and dried over Na2SO4. The solvent was removed under reduced pressure, and the residue was purified by flash chromatography eluting with EtOAc in hexane (0% to 70% gradient) to yield 5-bromo-8-methylquinoline as a yellow solid (360 mg, 30%). MS: m / z = 223.9 [M+H] + .

[0257] tert-Butyl N-[(3R,4R)-4-methyl-1-(8-methylquinolin-5-yl)pyrrolidin-3-yl]carbamate: To a solution of 5-bromo-8-methylquinoline (360 mg, 1.62 mmol) in N,N-dimethylformamide (10 mL) was added tert-butyl N-[(3R,4R)-4-methylpyrrolidin-3-yl]carbamate (342 mg, 1.71 mmol), Pd(dba)CHCl (177 mg, 0.17 mmol), KPO (1088 mg, 5.12 mmol), and Davephos (135 mg, 0.34 mmol) at room temperature. The resulting mixture was stirred for 130 minutes. o The mixture was stirred at RT for 3 h. When the reaction was complete, it was quenched by adding water (20 mL). The resulting mixture was extracted with DCM (100 mL x 3). The organic phases were combined, washed with brine, and dried over Na2SO4. The organic phases were combined, washed with brine, and dried over Na2SO4. The solvent was removed under reduced pressure, and the residue was purified by flash chromatography eluting with EtOAc in hexane (0% to 25% gradient) to afford tert-butyl N-[(3R,4R)-4-methyl-1-(8-methylquinolin-5-yl)pyrrolidin-3-yl]carbamate as a yellow oil (400 mg, 72%). MS: m / z = 342.2 [M+H] + .

[0258] (3R,4R)-4-Methyl-1-(8-methylquinolin-5-yl)pyrrolidin-3-amine: To a solution of tert-butyl N-[(3R,4R)-4-methyl-1-(8-methylquinolin-5-yl)pyrrolidin-3-yl]carbamate (200 mg, 0.59 mmol) in methanol (5 mL) was added hydrochloric acid solution (4 M in dioxane, 3 mL, 12 mmol) at room temperature. The resulting mixture was stirred at room temperature for 1 h. Upon completion of the reaction, the reaction mixture was concentrated under reduced pressure, and the residue was purified by prep-HPLC under the following conditions: Column: XBridge Shield Prep C18 OBD column, 150 mm, 5 μm; Mobile phase: acetonitrile in water (containing 10 mmol / L NH4HCO3 and 0.1% NH3.H2O), 5% to 60% gradient in 7 min; Detector: UV 254 nm. (3R,4R)-4-methyl-1-(8-methylquinolin-5-yl)pyrrolidin-3-amine was obtained as a yellow solid (60 mg, 42%).

[0259] Compound 33: HPLC: 99.5% purity, RT=1.07min. MS: m / z = 242.3 [M+H] + . 1 H NMR (300 MHz, methanol-d 4、 ppm) δ 8.82-8.74 (m, 1 H), 8.71-8.61 (m, 1 H), 7.48-7.37 (m, 2 H), 6.92 (d, J = 7.9 Hz, 1 H), 3.77-3.65 (m, 1 H), 3.62-3.51 (m, 1 H), 3.50-3.38 (m, 1 H), 3.37-3.25 (m, 1 H), 3.23-3.12 (m, 1 H), 2.63 (s, 3 H), 2.56-2.40 (m, 1 H), 1.14 (d, J = 7.0 Hz, 3 H). Example 14: Synthesis of Compound 34 (N-[(3R,4R)-4-methyl-1-(8-methylquinolin-5-yl)pyrrolidin-3-yl]-2-(1-methylpiperidin-4-yl)acetamide) [ka]

[0260] N-[(3R,4R)-4-Methyl-1-(8-methylquinolin-5-yl)pyrrolidin-3-yl]-2-(1-methylpiperidin-4-yl)acetamide: To a solution of (3R,4R)-4-methyl-1-(8-methylquinolin-5-yl)pyrrolidin-3-amine (40 mg, 0.16 mmol) in N-dimethylformamide (2 mL) was added 2-(1-methylpiperidin-4-yl)acetic acid (38 mg, 0.24 mmol), HATU (91 mg, 0.24 mmol), and DIEA (101 mg, 0.78 mmol) at room temperature. The resulting solution was stirred for 14 h. Upon completion of the reaction, the reaction mixture was concentrated under reduced pressure, and the residue was purified by prep-HPLC under the following conditions: column, XBridge Shield Prep C18 OBD column, 150 mm, 5 μm; mobile phase, acetonitrile in water (containing 10 mmol / L NH4HCO3 and 0.1% NH3.HO), 5% to 61% gradient in 7 min; detector, UV 254 nm. N-[(3R,4R)-4-methyl-1-(8-methylquinolin-5-yl)pyrrolidin-3-yl]-2-(1-methylpiperidin-4-yl)acetamide was obtained as a yellow solid (20 mg, 31%).

[0261] Compound 34: HPLC: 97.4% purity, RT=2.79min. MS: m / z = 381.4 [M+H] + . 1 H NMR (300 MHz, methanol-d 4、ppm) δ 8.84-8.75 (m, 1 H), 8.73-8.59 (m, 1 H), 7.50-7.39 (m, 2 H), 6.96 (d, J = 7.8 Hz, 1 H), 4.68-4.56 (m, 1 H), 3.80-3.68 (m, 1 H), 3.51-3.39 (m, 1 H), 3.32-3.19 (m, 1 H), 3.21-3.10 (m, 1 H), 2.90-2.78 (m, 2 H), 2.71-2.52 (m, 4 H), 2.30-2.12 (m, 5 H), 2.09-1.93 (m, 2 H), 1.88-1.62 (m, 3 H), 1.42-1.17 (m, 2 H), 1.06 (d, J = 7.0 Hz, 3 H).

[0262] The following compounds were synthesized in a similar manner:

[0263] Compound 39 (N-[(3R,4R)-4-methyl-1-(8-methylquinolin-5-yl)pyrrolidin-3-yl]-2-(morpholin-4-yl)acetamide): Purified from (3R,4R)-4-methyl-1-(8-methylquinolin-5-yl)pyrrolidin-3-amine and 2-(morpholin-4-yl)acetic acid on prep-HPLC under the following conditions: Column: XBridge Shield Prep C18 OBD column, 150 mm, 5 μm; Mobile phase: Acetonitrile in water (containing 10 mmol / L NH4HCO3 and 0.1% NH3.HO), 20% to 50% gradient in 8 min; Detector: UV 254 nm (20 mg, 24%, yellow solid). HPLC: 99.1% purity, RT = 1.29 min. MS: m / z = 369.4 [M+H] + . 1 H NMR (300 MHz, methanol-d 4、ppm) δ 8.85-8.76 (m, 1 H), 8.69-8.59 (m, 1 H), 7.52-7.41 (m, 2 H), 7.00 (d, J = 7.8 Hz, 1 H), 4.66-4.54 (m, 1 H), 3.81-3.64 (m, 5 H), 3.50-3.37 (m, 1 H), 3.31-3.11 (m, 3 H), 3.12-2.99 (m, 2 H), 2.78-2.57 (m, 4 H), 2.62-2.45 (m, 4 H), 1.08 (d, J = 7.0 Hz, 3 H).

[0264] Compound 40 (2-(1,4-dimethylpiperidin-4-yl)-N-[(3R,4R)-4-methyl-1-(8-methylquinolin-5-yl)pyrrolidin-3-yl]acetamide): Purified from (3R,4R)-4-methyl-1-(8-methylquinolin-5-yl)pyrrolidin-3-amine and 2-(1,4-dimethylpiperidin-4-yl)acetic acid by prep-HPLC under the following conditions: Column: XBridge Shield Prep C18 OBD column, 150 mm, 5 μm; Mobile phase: acetonitrile in water (containing 10 mmol / L NH4HCO3 and 0.1% NH3.HO), 30% to 60% gradient in 8 min; Detector: UV 254 nm (10 mg, 15%, pale yellow solid). HPLC: 97.3% purity, RT = 2.30 min. MS: m / z = 395.4 [M+H] + . 1 H NMR (300 MHz, DMSO-d 6、ppm) δ 8.83-8.74 (m, 1 H), 8.54-8.44 (m, 1 H), 7.83 (d, J = 8.3 Hz, 1 H), 7.42-7.31 (m, 2 H), 6.77 (d, J = 7.7 Hz, 1 H), 4.49-4.34 (m, 1 H), 3.68-3.57 (m, 1 H), 3.40-3.31 (m, 2 H), 3.23-3.11 (m, 1 H), 3.07-2.96 (m, 1 H), 2.55-2.49 (m, 4 H), 2.45-2.31 (m, 2 H), 2.23 (s, 3 H), 2.15-1.98 (m, 2 H), 1.60-1.47 (m, 2 H), 1.39-1.25 (m, 2 H), 1.20-1.13 (m, 1 H), 0.92-0.88 (m, 6 H).

[0265] Compound 47 ((2S)-2-hydroxy-3-methyl-N-[(3R,4R)-4-methyl-1-(8-methylquinolin-5-yl)pyrrolidin-3-yl]butanamide): Purified from (3R,4R)-4-methyl-1-(8-methylquinolin-5-yl)pyrrolidin-3-amine and (2S)-2-hydroxy-3-methybutanoic acid by prep-HPLC under the following conditions: column, XBridge Shield Prep C18 OBD column, 150 mm, 5 μm; mobile phase, acetonitrile in water (containing 10 mmol / L NH4HCO3 and 0.1% NH3.HO), 35% to 45% gradient in 7 min; detector, UV 254 nm (20 mg, 24%, orange solid). HPLC: 99.6% purity, RT=1.40min. MS: m / z = 342.4 [M+H] + . 1 H NMR (300 MHz, methanol-d 4、ppm) δ 8.85-8.77 (m, 1 H), 8.71-8.61 (m, 1 H), 7.53-7.42 (m, 2 H), 7.00 (d, J = 7.8 Hz, 1 H), 4.70-4.58 (m, 1 H), 3.94-3.86 (m, 1 H), 3.80-3.68 (m, 1 H), 3.53-3.40 (m, 1 H), 3.32-3.16 (m, 2 H), 2.76-2.60 (m, 4 H), 2.22-2.05 (m, 1 H), 1.10 (d, J = 7.0 Hz, 3 H), 1.03 (d, J = 7.0 Hz, 3 H), 0.92 (d, J = 6.8 Hz, 3H).

[0266] Compound 48 ((2R)-2-hydroxy-3-methyl-N-[(3R,4R)-4-methyl-1-(8-methylquinolin-5-yl)pyrrolidin-3-yl]butanamide): Purified from (3R,4R)-4-methyl-1-(8-methylquinolin-5-yl)pyrrolidin-3-amine and (2R)-2-hydroxy-3-methylbutanoic acid by prep-HPLC under the following conditions: Column: XBridge Shield Prep C18 OBD column, 150 mm, 5 μm; Mobile phase: acetonitrile in water (containing 10 mmol / L NH4HCO3 and 0.1% NH3.HO), 35% to 45% gradient in 7 min; Detector: UV 254 nm (23 mg, 29%, orange solid). HPLC: 99.7% purity, RT = 1.39 min. MS: m / z = 342.4 [M+H] + . 1 H NMR (300 MHz, methanol-d 4、ppm) δ 8.85-8.77 (m, 1 H), 8.70-8.60 (m, 1 H), 7.52-7.41 (m, 2 H), 7.01 (d, J = 7.8 Hz, 1 H), 4.69-4.56 (m, 1 H), 3.94 (d, J = 3.6 Hz, 1 H), 3.79-3.67 (m, 1 H), 3.53-3.41 (m, 1 H), 3.31-3.15 (m, 2 H), 2.77-2.62 (m, 4 H), 2.18-2.00 (m, 1 H), 1.10 (d, J = 7.0 Hz, 3 H), 1.01 (d, J = 6.9 Hz, 3 H), 0.84 (d, J = 6.8 Hz, 3 H). Example 15: Synthesis of compounds 35 and 36 (N-[(3R,4R)-4-methyl-1-(8-methylquinolin-5-yl)pyrrolidin-3-yl]-2-[(3S)-1-methylpiperidin-3-yl]acetamide and N-[(3R,4R)-4-methyl-1-(8-methylquinolin-5-yl)pyrrolidin-3-yl]-2-[(3R)-1-methylpiperidin-3-yl]acetamide) [ka]

[0267] N-[(3R,4R)-4-Methyl-1-(8-methylquinolin-5-yl)pyrrolidin-3-yl]-2-[(3S)-1-methylpiperidin-3-yl]acetamide and N-[(3R,4R)-4-Methyl-1-(8-methylquinolin-5-yl)pyrrolidin-3-yl]-2-[(3R)-1-methylpiperidin-3-yl]acetamide: To a solution of (3R,4R)-4-methyl-1-(8-methylquinolin-5-yl)pyrrolidin-3-amine (49 mg, 0.20 mmol) in N,N-dimethylformamide (3 mL) was added 2-(1-methylpiperidin-3-yl)acetic acid (46 mg, 0.30 mmol), HATU (120 mg, 0.30 mmol), and DIEA (126 mg, 0.98 mmol) at room temperature. The resulting solution was stirred at room temperature for 14 h. Upon completion, the reaction mixture was concentrated under reduced pressure, and the residue was first purified by prep-HPLC under the following conditions: column, XBridge Prep C18 OBD column, 150 mm, 5 μm; mobile phase, acetonitrile in water (containing 10 mmol / L NH4HCO3 and 0.1% NH3.H2O), 20% to 57% gradient in 7 min; detector, UV 254 nm. The two diastereomers were then separated by chiral prep-HPLC under the following conditions: column, CHIRALPAK IG-3, 0.46 x 10 cm, 3 μm; mobile phase, hexane in 80% isocratic EtOH (containing 0.1% DEA) in 30 min; detector, UV 254 nm.

[0268] Isomer 1: (15mg, 19%, red solid) HPLC: 99.8% purity, RT=1.31min. MS: m / z = 381.2 [M+H] + . 1 H NMR (300 MHz, methanol-d 4、ppm) δ 8.84-8.75 (m, 1 H), 8.72-8.62 (m, 1 H), 7.50-7.39 (m, 2 H), 6.96 (d, J = 7.8 Hz, 1 H), 4.68-4.56 (m, 1 H), 3.81-3.69 (m, 1 H), 3.51-3.39 (m, 1 H), 3.30-3.09 (m, 4 H), 2.72-2.51 (m, 8 H), 2.51-2.37 (m, 1 H), 2.31-2.09 (m, 3 H), 1.91-1.65 (m, 3 H), 1.24-1.10 (m, 1 H), 1.07 (d, J = 6.9 Hz, 3H).

[0269] Isomer 2: (18 mg, 10%, yellow solid) HPLC: 99.5% purity, RT=1.30min. MS:m / z=381.2[M+H] + . 1 H NMR (300 MHz, methanol-d 4、 ppm) δ 8.84-8.75 (m, 1 H), 8.73-8.63 (m, 1 H), 7.51-7.40 (m, 2 H), 6.96 (d, J = 7.8 Hz, 1 H), 4.69-4.56 (m, 1 H), 3.82-3.70 (m, 1 H), 3.51-3.35 (m, 2 H), 3.38-3.31 (m, 1 H), 3.30-3.21 (m, 1 H), 3.21-3.10 (m, 1 H), 2.83-2.69 (m, 4 H), 2.72-2.54 (m, 5 H), 2.38-2.21 (m, 3 H), 2.00-1.83 (m, 2 H), 1.86-1.65 (m, 1 H), 1.31-1.13 (m, 1 H), 1.07 (d, J = 6.9 Hz, 3 H).

[0270] The following compounds were synthesized in a similar manner:

[0271] Compounds 37 and 38 (N-[(3R,4R)-4-methyl-1-(8-methylquinolin-5-yl)pyrrolidin-3-yl]-2-[(3S)-1-methylpyrrolidin-3-yl]acetamide and N-[(3R,4R)-4-methyl-1-(8-methylquinolin-5-yl)pyrrolidin-3-yl]-2-[(3R)-1-methylpyrrolidin-3-yl]acetamide): were purified from (3R,4R)-4-methyl-1-(8-methylquinolin-5-yl)pyrrolidin-3-amine and 2-(1-methylpyrrolidin-3-yl)acetic acid on prep-HPLC under the following conditions: column, XBridge Prep C18 OBD column, 150 mm, 5 μm; mobile phase, acetonitrile in water (10 mmol / L). NH4HCO3 and 0.1% NH3.H2O), 20% to 47% gradient in 8 min; detector, UV 254 nm. The two isomers were separated on chiral prep-HPLC under the following conditions: column, CHIRALPAK IG-3, 0.46 x 10 cm, 3 um; mobile phase, 70% isocratic MtBE in EtOH (containing 0.1% DEA) in 30 min; detector, UV 254 nm. Isomer 1: (25 mg, 26%, red solid) HPLC: 97.7% purity, RT = 2.14 min. MS: m / z = 367.4 [M+H] + . 1 H NMR (400 MHz, DMSO-d 6、 ppm) δ 8.89-8.83 (m, 1 H), 8.60-8.48 (m, 1 H), 7.95 (d, J = 8.5 Hz, 1 H), 7.48-7.40 (m, 2 H), 6.84 (d, J = 7.8 Hz, 1 H), 4.54-4.43 (m, 1 H), 3.74-3.65 (m, 1 H), 3.50-3.35 (m, 1 H), 3.28-3.18 (m, 1 H), 3.13-3.02 (m, 1 H), 2.65-2.30 (m, 8 H), 2.27-2.05 (m, 6 H), 1.94-1.80 (m, 1 H), 1.41-1.28 (m, 1 H), 0.95 (d, J = 6.9Hz, 3H). Isomer 2: (25mg, 26%, yellow solid) HPLC: 95.2% purity, RT=2.16min. MS: m / z = 367.4 [M+H] + 。 1 H NMR (400 MHz、DMSO-d 6、 ppm) δ 8.89-8.83 (m、1 H)、8.60-8.53 (m、1 H)、7.96 (d、J = 8.5 Hz、1 H)、7.48-7.40 (m、2 H)、6.84 (d、J = 7.8 Hz、1 H)、4.54-4.43 (m、1 H)、3.74-3.65 (m、1 H)、3.44-3.35 (m、1 H)、3.28-3.19 (m、1 H)、3.12-3.04 (m、1 H)、2.62-2.57 (m、3 H)、2.57-2.50 (m、1 H)、2.49-2.37 (m、4 H)、2.20 (d、J = 9.5 Hz、5 H)、2.15-2.06 (m、1 H)、1.97-1.83 (m、1 H)、1.44-1.31 (m、1 H)、0.95 (d、J = 6.8 Hz、3 H)。

[0272] Compounds 45 and 46 ((2S)-2-cyclopropyl-2-hydroxy-N-[(3R,4R)-4-methyl-1-(8-methylquinolin-5-yl)pyrrolidin-3-yl]acetamide and (2R)-2-cyclopropyl-2-hydroxy-N-[(3R,4R)-4-methyl-1-(8-methylquinolin-5-yl)pyrrolidin-3-yl]acetamide): were purified from (3R,4R)-4-methyl-1-(8-methylquinolin-5-yl)pyrrolidin-3-amine and 2-cyclopropyl-2-hydroxyacetamide by prep-HPLC under the following conditions: column, XBridge RP18 OBD column, 150 mm, 5 μm; mobile phase, acetonitrile in water (containing 10 mmol / L NH4HCO3 and 0.1% NH3.H2O), 20% to 50% gradient in 7 min; detector, UV 254 nm. The two diastereomers were separated on chiral prep-HPLC under the following conditions: Column: CHIRALPAK IA-3, 0.46x5 cm, 3 um; Mobile phase: 70% isocratic hexane in EtOH (containing 0.1% DEA) in 15 min; Detector: UV 254 nm. Isomer 1: (25 mg, 22%, red solid) HPLC: 97.6% purity, RT=1.26 min. MS: m / z=340.3 [M+H] + . 1 H NMR (300 MHz, methanol-d 4、 ppm) δ 9.18-9.08 (m, 1 H), 8.92-8.83 (m, 1 H), 7.82-7.61 (m, 2 H), 7.04 (d, J = 8.1 Hz, 1 H), 4.68-4.56 (m, 1 H), 3.93-3.81 (m, 1 H), 3.67-3.54 (m, 2 H), 3.46-3.31 (m, 2 H), 2.77-2.61 (m, 4 H), 1.11-1.04 (m, 4 H), 0.56-0.29 (m, 4 H). Isomer 2: (25mg, 22%, red solid) HPLC: 98.1% purity, RT=1.26min. MS:m / z=340.4[M+H] + . 1 H NMR (300 MHz, methanol-d 4、ppm) δ 8.87-8.74 (m, 2 H), 7.59-7.48 (m, 2 H), 7.02 (d, J = 7.9 Hz, 1 H), 4.70-4.58 (m, 1 H), 3.83-3.72 (m, 1 H), 3.74-3.65 (m, 1 H), 3.57-3.44 (m, 1 H), 3.32-3.20 (m, 2 H), 2.77-2.62 (m, 4 H), 1.26-1.05 (m, 4 H), 0.60-0.37 (m, 4 H). Example 16: Synthesis of Compound 41 (2-(4-fluoropiperidin-4-yl)-N-[(3R,4R)-4-methyl-1-(8-methylquinolin-5-yl)pyrrolidin-3-yl]acetamide) [ka]

[0273] tert-Butyl 4-fluoro-4-([[(3R,4R)-4-methyl-1-(8-methylquinolin-5-yl)pyrrolidin-3-yl]carbamoyl]methyl)piperidine-1-carboxylate: To a solution of (3R,4R)-4-methyl-1-(8-methylquinolin-5-yl)pyrrolidin-3-amine (59 mg, 0.24 mmol) in N,N-dimethylformamide (3 mL) was added 2-[1-[(tert-butoxy)carbonyl]-4-fluoropiperidin-4-yl]acetic acid (101 mg, 0.39 mmol), HATU (138 mg, 0.36 mmol), and DIEA (153 mg, 1.18 mmol) at room temperature. The resulting solution was stirred at room temperature for 16 h. Upon completion of the reaction, the reaction mixture was concentrated under reduced pressure, and the residue was purified by prep-HPLC under the following conditions: column, XBridge Shield Prep C18 OBD column, 150 mm, 5 μm; mobile phase, acetonitrile in water (containing 10 mmol / L NH4HCO3 and 0.1% NH3.HO), 32% to 35% gradient in 7 min; detector, UV 254 nm. tert-Butyl 4-fluoro-4-([[(3R,4R)-4-methyl-1-(8-methylquinolin-5-yl)pyrrolidin-3-yl]carbamoyl]methyl)piperidine-1-carboxylate was obtained as a yellow solid (19 mg, 16%). MS: m / z=485.3 [M+H] + .

[0274] 2-(4-Fluoropiperidin-4-yl)-N-[(3R,4R)-4-methyl-1-(8-methylquinolin-5-yl)pyrrolidin-3-yl]acetamide: To a solution of tert-butyl 4-fluoro-4-([[(3R,4R)-4-methyl-1-(8-methylquinolin-5-yl)pyrrolidin-3-yl]carbamoyl]methyl)piperidine-1-carboxylate (17 mg, 0.04 mmol) in methanol (3 mL) was added hydrochloric acid solution (4 M in dioxane, 1 mL, 4 mmol) at room temperature. The resulting mixture was stirred at room temperature for 1 h. Upon completion of the reaction, the reaction mixture was concentrated under reduced pressure, and the residue was purified by prep-HPLC under the following conditions: column, XBridge Shield Prep C18 OBD column, 150 mm, 5 μm; mobile phase, acetonitrile in water (containing 10 mmol / L NH4HCO3 and 0.1% NH3.HO), 32% to 35% gradient in 7 min; detector, UV 254 nm. 2-(4-fluoropiperidin-4-yl)-N-[(3R,4R)-4-methyl-1-(8-methylquinolin-5-yl)pyrrolidin-3-yl]acetamide was obtained as a pale yellow solid (10 mg, 71%).

[0275] Compound 41: HPLC: 96.3% purity, RT=0.85min. MS:m / z=385.2[M+H] + . 1 H NMR (300 MHz, methanol-d 4、 ppm) δ 8.84-8.76 (m, 1 H), 8.74-8.63 (m, 1 H), 7.53-7.40 (m, 2 H), 7.01-6.92 (m, 1 H), 4.69-4.57 (m, 1 H), 3.82-3.70 (m, 1 H), 3.52-3.39 (m, 1 H), 3.36-3.00 (m, 6 H), 2.82-2.56 (m, 6 H), 2.23-1.85 (m, 4 H), 1.08 (d, J = 7.0 Hz, 3 H). Example 17: Synthesis of Compound 42 (2-(4-fluoro-1-methylpiperidin-4-yl)-N-[(3R,4R)-4-methyl-1-(8-methylquinolin-5-yl)pyrrolidin-3-yl]acetamide) [ka]

[0276] 2-(4-Fluoro-1-methylpiperidin-4-yl)-N-[(3R,4R)-4-methyl-1-(8-methylquinolin-5-yl)pyrrolidin-3-yl]acetamide: 2-(4-Fluoro-1-methylpiperidin-4-yl)-N-[(3R,4R)-4-methyl-1-(8-methylquinolin-5-yl)pyrrolidin-3-yl]acetamide (176 mg, 0.46 mmol) in MeOH (30 mL) (CHO) n (423 mg, 4.69 mmol) and NaOAc (790 mg, 9.64 mmol) were added at room temperature. The resulting mixture was stirred at room temperature for 2 h, and then NaBH (275 mg, 7.27 mmol) was added in portions. The reaction mixture was stirred at room temperature for another 16 h. When the reaction was complete, it was quenched by adding water (20 mL). The resulting mixture was extracted with DCM (50 mL x 3). The organic phases were combined, washed with brine, and dried over NaSO. The solvent was concentrated under reduced pressure, and the residue was purified by prep-HPLC under the following conditions: column, XBridge Shield Prep C18 OBD column, 150 mm, 5 μm; mobile phase, acetonitrile in water (containing 10 mmol / L NHHCO and 0.1% NH.H0), 30% to 43% gradient in 7 min; detector, UV 254 nm. 2-(4-Fluoro-1-methylpiperidin-4-yl)-N-[(3R,4R)-4-methyl-1-(8-methylquinolin-5-yl)pyrrolidin-3-yl]acetamide was obtained as a yellow solid (90 mg, 49%).

[0277] Compound 42: HPLC: 96.5% purity, RT=7.86min. MS:m / z=399.4[M+H] + . 1 H NMR (400 MHz, DMSO-d 6、ppm) δ 8.89-8.83 (m, 1 H), 8.60-8.52 (m, 1 H), 8.03 (d, J = 8.5 Hz, 1 H), 7.48-7.40 (m, 2 H), 6.84 (d, J = 7.8 Hz, 1 H), 4.55-4.43 (m, 1 H), 3.75-3.66 (m, 1 H), 3.45-3.36 (m, 1 H), 3.28-3.19 (m, 1 H), 3.14-3.06 (m, 1 H), 2.62-2.42 (m, 8 H), 2.18-2.03 (m, 5 H), 1.91-1.68 (m, 4 H), 0.97 (d, J = 6.8 Hz, 3 H). Example 18: Synthesis of compounds 43 and 44 (2-[(4S)-3,3-difluoro-1-methylpiperidin-4-yl]-N-[(3R,4R)-4-methyl-1-(8-methylquinolin-5-yl)pyrrolidin-3-yl]acetamide and 2-[(4R)-3,3-difluoro-1-methylpiperidin-4-yl]-N-[(3R,4R)-4-methyl-1-(8-methylquinolin-5-yl)pyrrolidin-3-yl]acetamide) [ka]

[0278] tert-Butyl 3,3-difluoro-4-([[(3R,4R)-4-methyl-1-(8-methylquinolin-5-yl)pyrrolidin-3-yl]carbamoyl]methyl)piperidine-1-carboxylate: To a solution of (3R,4R)-4-methyl-1-(8-methylquinolin-5-yl)pyrrolidin-3-amine (121 mg, 0.50 mmol) in N,N-dimethylformamide (3 mL) was added 2-{1-[(tert-butoxy)carbonyl]-3,3-difluoropiperidin-4-yl}acetic acid (363 mg, 1.30 mmol), HATU (342 mg, 0.90 mmol), and DIEA (380 mg, 2.94 mmol) at room temperature. The resulting solution was stirred at room temperature for 2 h. When the reaction was complete, it was quenched by adding water (20 mL). The resulting mixture was extracted with DCM (60 mL x 3). The organic phases were combined, washed with brine, and dried over Na2SO4. The solvent was removed under reduced pressure to yield tert-butyl 3,3-difluoro-4-([[(3R,4R)-4-methyl-1-(8-methylquinolin-5-yl)pyrrolidin-3-yl]carbamoyl]methyl)piperidine-1-carboxylate as a yellow oil (300 mg, crude). MS: m / z = 503.4 [M+H] + .

[0279] 2-(3,3-Difluoropiperidin-4-yl)-N-[(3R,4R)-4-methyl-1-(8-methylquinolin-5-yl)pyrrolidin-3-yl]acetamide: To a solution of tert-butyl 3,3-difluoro-4-([[(3R,4R)-4-methyl-1-(8-methylquinolin-5-yl)pyrrolidin-3-yl]carbamoyl]methyl)piperidine-1-carboxylate (300 mg, crude product) in methanol (4 mL) was added hydrochloric acid solution (4 M in dioxane, 4 mL, 16 mmol) at room temperature. The resulting mixture was stirred at room temperature for 2 h. When the reaction was complete, the pH value of the reaction mixture was adjusted to 7-8 using saturated NaHCO3 solution. The resulting mixture was extracted with ethyl acetate (50 mL x 3). The organic phases were combined, washed with brine, and dried over Na2SO4. The solvent was removed under reduced pressure to yield 2-(3,3-difluoropiperidin-4-yl)-N-[(3R,4R)-4-methyl-1-(8-methylquinolin-5-yl)pyrrolidin-3-yl]acetamide as a yellow oil (330 mg, crude). MS: m / z=403.4 [M+H] + .

[0280] 2-[(4S)-3,3-Difluoro-1-methylpiperidin-4-yl]-N-[(3R,4R)-4-methyl-1-(8-methylquinolin-5-yl)pyrrolidin-3-yl]acetamide and 2-[(4R)-3,3-Difluoro-1-methylpiperidin-4-yl]-N-[(3R,4R)-4-methyl-1-(8-methylquinolin-5-yl)pyrrolidin-3-yl]acetamide: To a solution of 2-(3,3-difluoropiperidin-4-yl)-N-[(3R,4R)-4-methyl-1-(8-methylquinolin-5-yl)pyrrolidin-3-yl]acetamide (330 mg, crude) in methanol (50 mL) (CHO) n(684 mg, 7.59 mmol), NaOAC (1.27 g, 15.52 mmol) were added at room temperature. The resulting mixture was stirred at room temperature for 2 h, and then NaBH4 (444 mg, 11.73 mmol) was added in portions. The reaction mixture was stirred at room temperature for another 16 h. When the reaction was complete, it was quenched by adding water (20 mL). The resulting mixture was extracted with DCM (50 mL x 3). The organic phases were combined, washed with brine, and dried over Na2SO4. The solvent was removed under reduced pressure, and the residue was first purified by prep-HPLC under the following conditions: column, XBridge Prep C18 OBD column, 150 mm, 5 μm; mobile phase, acetonitrile in water (containing 10 mmol / L NH4HCO3 and 0.1% NH3.H2O), 27% to 46% gradient in 7 min; detector, UV 254 nm. The two diastereomers were then separated on chiral prep-HPLC under the following conditions: column, Repaired ADH, 0.46 x 10 cm, 3 μm; mobile phase, hexane in 70% isocratic EtOH (containing 0.1% DEA) in 30 min; detector, UV 254 nm.

[0281] Isomer 1: (15 mg, 7% in 3 steps, off-white solid) HPLC: 97.9% purity, RT=2.77min. MS:m / z=209.1[M+H] + . 1 H NMR (300 MHz, methanol-d 4、 ppm) δ 8.83-8.75 (m, 1 H), 8.73-8.59 (m, 1 H), 7.50-7.39 (m, 2 H), 6.96 (d, J = 7.8 Hz, 1 H), 4.68-4.56 (m, 1 H), 3.80-3.68 (m, 1 H), 3.56-3.39 (m, 1 H), 3.29-2.97 (m, 3 H), 2.87-2.54 (m, 6 H), 2.40-2.18 (m, 6 H), 2.16-2.01 (m, 1 H), 1.81-1.69 (m, 1 H), 1.61-1.42 (m, 1 H), 1.06 (d, J = 6.9 Hz, 3H).

[0282] Isomer 2: (15 mg, 7% in 3 steps, off-white solid) HPLC: 98.6% purity, RT=2.70min. MS:m / z=209.0[M+H] + . 1 H NMR (300 MHz, methanol-d 4、 ppm) δ 8.84-8.75 (m, 1 H), 8.74-8.64 (m, 1 H), 7.51-7.40 (m, 2 H), 6.97 (d, J = 7.8 Hz, 1 H), 4.70-4.54 (m, 1 H), 3.79-3.67 (m, 1 H), 3.52-3.33 (m, 1 H), 3.31-2.98 (m, 3 H), 2.91-2.81 (m, 1 H), 2.77-2.55 (m, 5 H), 2.41-2.20 (m, 6 H), 2.20-2.06 (m, 1 H), 1.91-1.80 (m, 1 H), 1.66-1.52 (m, 1 H), 1.06 (d, J = 6.9 Hz, 3 H). Example 19: Synthesis of compounds 49 and 50 ((3S,4R)-4-fluoro-1-(8-trifluoromethyl-quinolin-5-yl)-pyrrolidin-3-ylamine and (3R,4S)-4-fluoro-1-(8-trifluoromethyl-quinolin-5-yl)-pyrrolidin-3-ylamine) [ka]

[0283] [cis-4-Fluoro-1-(8-trifluoromethyl-quinolin-5-yl)-pyrrolidin-3-yl]-carbamic acid tert-Butyl ester: To a solution of 5-bromo-8-(trifluoromethyl)quinoline (400 mg; 1.45 mmol) in tBuOH (9.0 mL) was added cis-(3-boc-amino)-4-fluoropyrrolidine (355 mg; 1.74 mmol), chloro(2-dicyclohexylphosphino-2',6'-di-i-propoxy-1,1'-biphenyl)[2-(2-aminoethylphenyl)]palladium(II), methyl t-butyl ether adduct (59.2 mg; 0.072 mmol), 2-dicyclohexylphosphino-2',6'-di-i-propoxy-1,1'-biphenyl (33.8 mg; 0.072 mmol), and cesium carbonate (944 mg; 2.90 mmol). The resulting mixture was sparged with nitrogen for 10 min and then 85 o The reaction mixture was microwaved at 17°C for 8 h. The reaction mixture was concentrated under reduced pressure, suspended in DCM (20 mL), sonicated for 30 s, and filtered through Celite. The solvent was removed under reduced pressure, and the residue was purified by flash chromatography eluting with EtOAc in hexanes (10% to 80% gradient) to afford [cis-4-fluoro-1-(8-trifluoromethyl-quinolin-5-yl)-pyrrolidin-3-yl]-carbamic acid tert-butyl ester as a light brown glassy solid (507 mg; 88%). MS: m / z = 400 [M+H] + .

[0284] (3S,4R)-4-Fluoro-1-(8-trifluoromethyl-quinolin-5-yl)-pyrrolidin-3-ylamine and (3R,4S)-4-Fluoro-1-(8-trifluoromethyl-quinolin-5-yl)-pyrrolidin-3-ylamine: To a solution of [cis-4-fluoro-1-(8-trifluoromethyl-quinolin-5-yl)-pyrrolidin-3-yl]-carbamic acid tert-butyl ester (485 mg; 1.21 mmol) in methanol (12 mL) was added hydrochloric acid solution (4 M in dioxane, 9 mL, 36 mmol) at room temperature. The resulting mixture was stirred overnight at room temperature. When the reaction was complete, ether (40 mL) was added to the orange solution, and the resulting yellow suspension was stirred at room temperature for 1 h. The solid was filtered and dissolved in 5N potassium hydroxide solution. The mixture was extracted with ethyl acetate, and the combined organic phase was washed with brine (30 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The two enantiomers were separated by chiral prep-HPLC under the following conditions: column, Lux Cellulose-2, 250 x 21.20 mm, 5 μm; mobile phase, 30% isocratic EtOH containing 0.5% DMEA in 11 min; detector, UV 254 nm.

[0285] Isomer 1: (86 mg, 23%, light brown solid) HPLC: 98.9% purity, RT=1.78min. MS:m / z=300[M+H] + . 1 H NMR (400 MHz, chloroform-d, ppm) d 8.86 (d, J = 1.7 Hz, 1H), 8.64 (d, J = 5.6 Hz, 1H), 8.60 (d, J = 1.7 Hz, 1H), 6.49 (d, J = 5.7 Hz, 1H), 6.41 (s, 1H), 4.29-4.14 (m, 2H), 4.08 (dt, J = 12.2, 7.3 Hz, 1H), 3.91 (q, J = 9.2, 8.7 Hz, 1H), 3.40-3.26 (m, 2H), 3.06 (p, J = 7.6 Hz, 1H), 2.56 (t, J = 6.0 Hz, 2H), 2.52 (q, J = 7.1 Hz, 4H), 2.41-2.24 (m, 2H), 0.99 (t, J = 7.1 Hz, 6H).

[0286] Isomer 2: (104mg, 29%, cream solid) HPLC:>99.9% purity, RT=1.75min. MS:m / z=300[M+H] + . 1 H NMR (400 MHz, chloroform-d, ppm) d 8.99 (dd, J = 4.1, 1.7 Hz, 1H), 8.47 (dd, J = 8.7, 1.7 Hz, 1H), 7.90 (dd, J = 8.4, 0.8 Hz, 1H), 7.37 (dd, J = 8.7, 4.1 Hz, 1H), 6.75 (d, J = 8.3 Hz, 1H), 5.05 (dt, J = 54.5, 3.4 Hz, 1H), 4.05 (ddd, J = 37.9, 12.3, 3.6 Hz, 1H), 3.79 - 3.54 (m, 4H), 1.52 (s, 2H). Example 20: Synthesis of Compound 51 ((3R,4R)-4-methyl-1-(8-methyl-[1,7]naphthyridin-5-yl)-pyrrolidin-3-ylamine hydrochloride) [ka]

[0287] [(3R,4R)-4-Methyl-1-(8-methyl-[1,7]naphthyridin-5-yl)-pyrrolidin-3-yl]-carbamic acid tert-butyl ester: To a solution of 5-bromo-8-methyl-[1,7]naphthyridine (100 mg; 0.45 mmol) in tBuOH (1.5 mL) was added (3R,4R)-3-(boc-amino)-4-methylpyrrolidine (104 mg; 0.49 mmol), chloro(2-dicyclo[2.0]phenyl]pyrrolidine (2.0 mL), and tert-butyl ester. hexylphosphino-2',6'-di-i-propoxy-1,1'-biphenyl)[2-(2-aminoethylphenyl)]palladium(II), methyl t-butyl ether adduct (18.3 mg; 0.02 mmol), 2-dicyclohexylphosphino-2',6'-di-i-propoxy-1,1'-biphenyl (10.5 mg; 0.02 mmol), and cesium carbonate (292 mg; 0.90 mmol) were added. The resulting mixture was sparged with nitrogen for 10 minutes and then cooled to 100°C. oThe mixture was microwaved at 17°C for 8 h. The reaction mixture was concentrated under reduced pressure, and the residue was purified by flash chromatography eluting with EtOAc in hexanes (20% to 80% gradient) to afford [(3R,4R)-4-methyl-1-(8-methyl-[1,7]naphthyridin-5-yl)-pyrrolidin-3-yl]-carbamic acid tert-butyl ester as a pale yellow solid (70 mg; 46%). MS: m / z=343 [M+H] + .

[0288] (3R,4R)-4-Methyl-1-(8-methyl-[1,7]naphthyridin-5-yl)-pyrrolidin-3-ylamine hydrochloride: To a solution of [(3R,4R)-4-methyl-1-(8-methyl-[1,7]naphthyridin-5-yl)-pyrrolidin-3-yl]-carbamic acid tert-butyl ester (120 mg, 0.35 mmol) in methanol (3 mL) was added hydrochloric acid solution (4 M in dioxane, 1.3 mL, 5.3 mmol) at room temperature. The resulting mixture was stirred overnight at room temperature. Upon completion of the reaction, the mixture was concentrated to dryness to yield (3R,4R)-4-methyl-1-(8-methyl-[1,7]naphthyridin-5-yl)-pyrrolidin-3-ylamine hydrochloride as a yellow amorphous solid (110 mg; 97%).

[0289] Compound 51: HPLC: 98.9% purity, RT=0.93min. MS:m / z=243[M+H] + . Example 21: Synthesis of compound 52 (N-[(3R,4R)-4-methyl-1-(8-methyl-[1,7]naphthyridin-5-yl)-pyrrolidin-3-yl]-2-(1-methyl-piperidin-4-yl)-acetamide) [ka]

[0290] N-[(3R,4R)-4-Methyl-1-(8-methyl-[1,7]naphthyridin-5-yl)-pyrrolidin-3-yl]-2-(1-methyl-piperidin-4-yl)-acetamide: To a solution of (3R,4R)-4-methyl-1-(8-methyl-[1,7]naphthyridin-5-yl)-pyrrolidin-3-ylamine hydrochloride (30 mg, 0.1 mmol) in N,N-dimethylformamide (1 mL) was added 2-(1-methylpiperidin-4-yl)acetic acid (16.5 mg, 0.105 mmol), DIEA (63 mL, 0.948 mmol), and (benzotriazol-1-yloxy)tris(dimethylamino)phosphonium hexafluorophosphate (51 mg, 0.114 mmol). The resulting solution was stirred at room temperature for 1 hour. Upon completion of the reaction, the solvent was removed under reduced pressure, and the residue was purified by prep-HPLC under the following conditions: column, XBridge Prep C18 OBD 10 mm, column, 30 x 250 mm; mobile phase, acetonitrile in water (containing 0.1% NH4OH), 10% to 60% gradient in 15 min; detector, UV 254 nm. N-[(3R,4R)-4-methyl-1-(8-methyl-[1,7]naphthyridin-5-yl)-pyrrolidin-3-yl]-2-(1-methyl-piperidin-4-yl)-acetamide was obtained as a white amorphous solid (32 mg, 88%).

[0291] Compound 52: HPLC:>99% purity, RT=1.30min. MS:m / z=382[M+H] + . 1H NMR (400 MHz, DMSO-d6, ppm) d 8.96 (dd, J = 4.1, 1.6 Hz, 1H), 8.60 (dd, J = 8.7, 1.6 Hz, 1H), 7.91 (d, J = 8.4 Hz, 1H), 7.86 (s, 1H), 7.66 (dd, J = 8.7, 4.1 Hz, 1H), 4.48 (dtd, J = 9.2, 6.0, 3.4 Hz, 1H), 3.85 (dd, J = 9.9, 6.0 Hz, 1H), 3.51 (dd, J = 9.1, 7.3 Hz, 1H), 3.36 (d, J = 8.9 Hz, 1H), 3.20 (dd, J = 9.9, 3.4 Hz, 1H), 2.81 (s, 3H), 2.73 - 2.61 (m, 2H), 2.49 - 2.43 (m, 1H), 2.10 (s, 3H), 2.08 - 1.99 (m, 2H), 1.77 (tdd, J = 11.3, 5.8, 2.4 Hz, 2H), 1.65 - 1.45 (m, 3H), 1.20 - 1.05 (m, 2H), 0.97 (d, J = 6.8 Hz, 3H).

[0292] The following compounds were synthesized in a similar manner:

[0293] Compound 53 (2-(1-methyl-azetidin-3-yl)-N-[(3R,4R)-4-methyl-1-(8-methyl-[1,7]naphthyridin-5-yl)-pyrrolidin-3-yl]-acetamide): Purified from 8-[(3R,4R)-3-amino-4-methylpyrrolidin-1-yl]quinoxaline-5-carbonitrile and (1-methyl-azetidin-3-yl)-acetic acid hydrochloride by prep-HPLC under the following conditions: column, XBridge Prep C18 OBD 10 μm, column, 30 × 250 mm; mobile phase, acetonitrile in water (containing 0.1% NH4OH), 10% to 60% gradient in 15 min; detector, UV 254 nm (29 mg, 86%, white amorphous solid). HPLC: >99% purity, RT = 1.24 min. MS: m / z=354 [M+H] + . 1H NMR (400 MHz, DMSO-d6, ppm) d 8.97 (dd, J = 4.1, 1.5 Hz, 1H), 8.61 (dd, J = 8.7, 1.7 Hz, 1H), 7.97 (d, J = 8.5 Hz, 1H), 7.87 (s, 1H), 7.67 (dd, J = 8.7, 4.1 Hz, 1H), 4.46 (dtd, J = 9.3, 6.0, 3.4 Hz, 1H), 3.85 (dd, J = 9.9, 6.0 Hz, 1H), 3.51 (dd, J = 9.1, 7.3 Hz, 1H), 3.38 - 3.32 (m, 1H), 3.28 - 3.15 (m, 3H), 2.82 (s, 3H), 2.78 - 2.68 (m, 2H), 2.61 - 2.55 (m, 1H), 2.48 - 2.31 (m, 3H), 2.14 (s, 3H), 0.95 (d, J = 6.9 Hz, 3H). Example 22: Synthesis of Compound 54 (1-pyrido[2,3-b]pyrazin-8-yl-pyrrolidine-3-carboxylic acid (2-diethylamino-ethyl)-amide) [ka]

[0294] 3-(2-Diethylamino-ethylcarbamoyl)-pyrrolidine-1-carboxylic acid tert-butyl ester: To a solution of 1-boc-pyrrolidine-3-carboxylic acid (1.0 g, 4.65 mmol) in anhydrous dichloromethane (25.0 mL) was added N,N-diethylethylenediamine (653 μL, 4.65 mmol), DIEA (4.0 mL, 23.2 mmol), and a 50% solution of propylphosphonic anhydride (8.2 mL, 13.9 mmol) in ethyl acetate. The resulting solution was stirred at room temperature for 1 h 30 h. Upon completion of the reaction, it was diluted with dichloromethane (70 mL), extracted with saturated sodium bicarbonate solution (3×50 mL), and washed with brine (50 mL). The organic phase was dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and the residue was purified by flash chromatography using an NH2-bonded silica column eluted with EtOAC in hexane (10% to 60% gradient) to yield 3-(2-diethylamino-ethylcarbamoyl)-pyrrolidine-1-carboxylic acid tert-butyl ester as a colorless oil (1.21 g, 83%). MS: m / z = 314 [M+H] + .

[0295] Pyrrolidine-3-carboxylic acid (2-diethylamino-ethyl)-amide hydrochloride: To a solution of 3-(2-diethylamino-ethylcarbamoyl)-pyrrolidine-1-carboxylic acid tert-butyl ester (1.2 g; 3.73 mmol) in methanol (30 mL) was added hydrochloric acid solution (4 M in dioxane, 28 mL, 112 mmol) at room temperature. The resulting mixture was stirred at room temperature overnight. When the reaction was complete, the mixture was concentrated under reduced pressure, the residue was dissolved in isopropanol (20 mL), acetonitrile (50 mL) and ether (10 mL) were added, and the colorless solution was stirred at room temperature overnight. The white suspension was filtered and the white solid was dried under vacuum to yield pyrrolidine-3-carboxylic acid (2-diethylamino-ethyl)-amide hydrochloride as a white solid (581 mg, 62%). MS: m / z=214 [M+H] + .

[0296] 1-Pyrido[2,3-b]pyrazin-8-yl-pyrrolidine-3-carboxylic acid (2-diethylamino-ethyl)-amide: A solution of 8-chloro-pyrido[2,3-b]pyrazine (60 mg, 0.362 mmol), pyrrolidine-3-carboxylic acid (2-diethylamino-ethyl)-amide hydrochloride (136 mg, 0.544 mmol) and DIEA (316 μl, 1.81 mmol) in absolute ethanol (3 mL) was added to 100 mL of ethanol. o C for 3 h. When the reaction was complete, the solvent was removed under reduced pressure, and the residue was purified by flash chromatography using an NH-bonded silica column eluting with methanol in dichloromethane (0% to 7% gradient) to afford 1-pyrido[2,3-b]pyrazin-8-yl-pyrrolidine-3-carboxylic acid (2-diethylamino-ethyl)-amide as a yellow solid (108 mg, 85%).

[0297] Compound 54: HPLC: 97.3% purity, RT=1.00min. MS:m / z=343[M+H] + . 1 H NMR (400 MHz, chloroform-d, ppm) d 8.86 (d, J = 1.7 Hz, 1H), 8.64 (d, J = 5.6 Hz, 1H), 8.60 (d, J = 1.7 Hz, 1H), 6.49 (d, J = 5.7 Hz, 1H), 6.41 (s, 1H), 4.29-4.14 (m, 2H), 4.08 (dt, J = 12.2, 7.3 Hz, 1H), 3.91 (q, J = 9.2, 8.7 Hz, 1H), 3.40-3.26 (m, 2H), 3.06 (p, J = 7.6 Hz, 1H), 2.56 (t, J = 6.0 Hz, 2H), 2.52 (q, J = 7.1 Hz, 4H), 2.41-2.24 (m, 2H), 0.99 (t, J = 7.1 Hz, 6H). Example 23: Synthesis of Compound 55 ([1-(1,2-dimethyl-1H-pyrrolo[2,3-b]pyridin-4-yl)-pyrrolidin-3-ylmethyl]-(2-piperidin-1-yl-ethyl)-amine) [ka]

[0298] 3-[(2-Piperidin-1-yl-ethylamino)-methyl]-pyrrolidine-1-carboxylic acid tert-butyl ester: To a solution of 3-formyl-pyrrolidine-1-carboxylic acid tert-butyl ester (200 mg, 0.98 mmol) and 2-piperidin-1-yl-ethylamine (126 mg, 0.98 mmol) in a mixture of methanol (0.40 mL) and 1,2-dichloroethane (1.60 mL), acetic acid (66 mg, 1.08 mmol) was added, and the reaction mixture was stirred at room temperature for 1 h. Sodium triacetoxyborohydride (329 mg, 1.48 mmol) was then added, and the reaction mixture was stirred at room temperature for 24 h. The reaction mixture was then poured into ice-cold water (10 mL) and extracted with ethyl acetate. The combined organic phases were dried over anhydrous sodium sulfate, filtered and concentrated to yield 3-[(2-piperidin-1-yl-ethylamino)-methyl]-pyrrolidine-1-carboxylic acid tert-butyl ester as a pale yellow gum (150 mg, 38%). MS: m / z=312.3 [M+H] + .

[0299] 3-{[benzyloxycarbonyl-(2-piperidin-1-yl-ethyl)amino]-methyl}-pyrrolidine-1-carboxylic acid tert-butyl ester: To a solution of 3-[(2-piperidin-1-yl-ethylamino)-methyl]-pyrrolidine-1-carboxylic acid tert-butyl ester (150 mg, 0.38 mmol) in THF (1.2 mL) and water (0.3 mL) was added potassium hydroxide (45 mg, 1.13 mmol) and 50% benzyl chloroformate in toluene (0.12 mL, 0.41 mmol). The reaction mixture was stirred at room temperature for 24 h, poured into ice-cold water (10 mL), and extracted with ethyl acetate. The combined organic phase was dried over anhydrous sodium sulfate, filtered and concentrated to give 3-{[benzyloxycarbonyl-(2-piperidin-1-yl-ethyl)-amino]-methyl}-pyrrolidine-1-carboxylic acid tert-butyl ester as a pale yellow gum (150 mg, 47%). MS: m / z=446.3 [M+H] + .

[0300] (2-Piperidin-1-yl-ethyl)-pyrrolidin-3-ylmethyl-carbamic acid benzyl ester trifluoroacetate: 3-{[benzyloxycarbonyl-(2-piperidin-1-yl-ethyl)-amino]-methyl}-pyrrolidine-1-carboxylic acid tert-butyl ester (150 mg, 0.18 mmol) o To a solution of dichloromethane (0.75 mL) at C was added trifluoroacetic acid (0.75 mL) dropwise over a period of 30 minutes. The reaction mixture was stirred at room temperature for 5 h. The reaction mixture was then concentrated under reduced pressure and azeotroped with toluene to yield (2-piperidin-1-yl-ethyl)-pyrrolidin-3-ylmethyl-carbamic acid benzyl ester trifluoroacetate as a brown gum (80 mg, 57%). MS: m / z=346.3 [M+H] + .

[0301] [1-(1,2-Dimethyl-1H-pyrrolo[2,3-b]pyridin-4-yl)-pyrrolidin-3-ylmethyl]-(2-piperidin-1-yl-ethyl)-carbamic acid benzyl ester: To a solution of 4-bromo-1,2-dimethyl-1H-pyrrolo[2,3-b]pyridine (200 mg, 0.88 mmol) and (2-piperidin-1-yl-ethyl)-pyrrolidin-3-ylmethyl-carbamic acid benzyl ester trifluoroacetate (847 mg, 1.06 mmol) in toluene (2 mL) was added potassium tert-butoxide (307 mg, 2.65 mmol), 2,2′-bis-diphenylphosphanyl-[1,1′]binaphthalenyl (56 mg, 0.09 mmol). The reaction mixture was degassed for 30 min before adding tris(dibenzylideneacetone)dipalladium(0) (42 mg, 0.04 mmol). The reaction mixture was then stored in a sealed tube for 24 h at 100°C. oThe mixture was heated at RT, poured into ice-cold water (30 mL) and extracted with ethyl acetate. The combined organic phases were dried over anhydrous sodium sulfate, filtered and concentrated to afford [1-(1,2-dimethyl-1H-pyrrolo[2,3-b]pyridin-4-yl)-pyrrolidin-3-ylmethyl]-(2-piperidin-1-yl-ethyl)-carbamic acid benzyl ester as a black gum (220 mg, 15%). MS: m / z=490.2 [M+H] + .

[0302] [1-(1,2-Dimethyl-1H-pyrrolo[2,3-b]pyridin-4-yl)-pyrrolidin-3-ylmethyl]-(2-piperidin-1-yl-ethyl)-amine: To a solution of [1-(1,2-dimethyl-1H-pyrrolo[2,3-b]pyridin-4-yl)-pyrrolidin-3-ylmethyl]-(2-piperidin-1-yl-ethyl)-carbamic acid benzyl ester (200 mg, 0.12 mmol) in dioxane (1 mL) was added 4 M hydrochloric acid solution in dioxane (0.15 mL, 0.60 mmol). The resulting pink reaction mixture was 65 o The mixture was heated at RT for 5 h and concentrated under reduced pressure. The resulting mixture was dissolved in water, basified with aqueous sodium bicarbonate solution, and extracted with ethyl acetate. The combined organic phases were dried over anhydrous sodium sulfate, filtered, and concentrated to provide the crude product, which was purified by flash chromatography to yield [1-(1,2-dimethyl-1H-pyrrolo[2,3-b]pyridin-4-yl)-pyrrolidin-3-ylmethyl]-(2-piperidin-1-yl-ethyl)-amine as a brown gum (10 mg, 23%).

[0303] Compound 55: HPLC: 97.3% purity, RT=1.00min. MS:m / z=343[M+H] + . 1H NMR (400 MHz, chloroform-d, ppm) d 8.86 (d, J = 1.7 Hz, 1H), 8.64 (d, J = 5.6 Hz, 1H), 8.60 (d, J = 1.7 Hz, 1H), 6.49 (d, J = 5.7 Hz, 1H), 6.41 (s, 1H), 4.29-4.14 (m, 2H), 4.08 (dt, J = 12.2, 7.3 Hz, 1H), 3.91 (q, J = 9.2, 8.7 Hz, 1H), 3.40-3.26 (m, 2H), 3.06 (p, J = 7.6 Hz, 1H), 2.56 (t, J = 6.0 Hz, 2H), 2.52 (q, J = 7.1 Hz, 4H), 2.41-2.24 (m, 2H), 0.99 (t, J = 7.1 Hz, 6H). Example 24: HEK TLR7 cell assay

[0304] 5000 c / w TLR7 / NFKb HEK cells were plated in 30 μL of phenol red-free DMEM (Gibco #31053) containing 10% FCS-HI, 2 mM L-Glutamine, and 1% Pen / Strept in a 384-well culture plate (Corning 3707). Cells were incubated at 37°C and 10% carbon dioxide for 24 h. 3 μL of control, standard, and compound were dispensed into wells and incubated for 30 minutes, followed by the addition of 3 μL of R848 agonist (10 μM final concentration) in 20 mM Hepes. The wells were incubated for 5 hours, followed by 15 minutes at room temperature. 10 μL of Steady-Glo substrate reagent was added, and the assay plate was rocked at 1500 rpm for 5 minutes. The assay plate was allowed to sit at room temperature for 30 minutes, after which the plate was read on the EnVision. Example 25: HEK TLR8 Cell Assay

[0305] 5000 c / w TLR7 / NFKb HEK cells were placed in 30 μL of phenol red-free DMEM (Gibco #31053) containing 10% FCS-HI, 2 mM L-Glutamine, and 1% Pen / Strept in a 384-well culture plate (Corning 3707). Cells were incubated at 37°C and 10% carbon dioxide for 24 h. 3 μL of control, standard, and compound were dispensed into wells and incubated for 30 minutes, followed by the addition of 3 μL of R848 agonist (30 μM final concentration) in 20 mM Hepes. The wells were incubated for 5 hours, followed by a 15-minute incubation at room temperature. 10 μL of Steady-Glo substrate reagent was added, and the assay plate was rocked at 1500 rpm for 5 minutes. The assay plate was allowed to sit at room temperature for 30 minutes, after which the plate was read on the EnVision.

[0306] The results are given in the table below. A:IC 50 <1uM B:IC 50 :1uM-10uM C:IC 50 >10uM Table 2 [Table 15] [Table 16] [Table 17] Example 26 Pharmaceutical Preparations

[0307] (A) Injection vials: A solution of 100 g of the active ingredient according to the invention and 5 g of disodium hydrogen phosphate in double-distilled water is adjusted to pH 6.5 using 2N hydrochloric acid, sterile filtered, transferred into injection vials, lyophilized under sterile conditions and sealed under sterile conditions, each injection vial containing 5 mg of the active ingredient.

[0308] (B) Suppositories: A mixture of 20 g of an active ingredient according to the invention is melted with 100 g of soybean lecithin and 1400 g of cocoa butter, poured into moulds and allowed to cool. Each suppository contains 20 mg of the active ingredient.

[0309] (C) Solution: A solution is prepared from 1 g of the active ingredient according to the invention, 9.38 g of NaH2PO4·2H2O, 28.48 g of Na2HPO4·12H2O and 0.1 g of benzalkonium chloride in 940 ml of double-distilled water. The pH is adjusted to 6.8, the solution is made up to 1 liter and sterilized by radiation. This solution can be used in the form of eye drops.

[0310] (D) Ointment: 500 mg of an active ingredient according to the invention is mixed with 99.5 g of petrolatum under aseptic conditions.

[0311] (E) Tablets: A mixture of 1 kg of active ingredient according to the invention, 4 kg of lactose, 1.2 kg of potato starch, 0.2 kg of talc and 0.1 kg of magnesium stearate is compressed in the conventional manner to give tablets, each tablet containing 10 mg of the active ingredient.

[0312] (F) Coated tablets: Tablets are compressed similarly to Example E and then coated in the conventional manner with a coating of sucrose, potato starch, talc, tragacanth and dye.

[0313] (G) Capsules: 2 kg of the active ingredient according to the invention are introduced into hard gelatin capsules in the conventional manner so that each capsule contains 20 mg of the active ingredient.

[0314] (H) Ampoules: A solution of 1 kg of the active ingredient according to the invention in 60 l of double-distilled water is sterile-filtered into ampoules, lyophilized under sterile conditions and sealed under sterile conditions, each ampoule containing 10 mg of the active ingredient.

[0315] (I) Inhalation spray: 14 g of the active ingredient according to the invention is dissolved in 10 liters of isotonic NaCl solution and the solution is transferred into a commercially available spray container with a pump mechanism. The solution can be sprayed into the mouth or nose. One spray (approximately 0.1 ml) corresponds to a dose of approximately 0.14 mg.

[0316] While numerous embodiments of the present invention have been described herein, it will be apparent that the basic examples may be modified to provide other embodiments that utilize the compounds and methods of the present invention. Therefore, it will be appreciated that the scope of this invention is to be defined by the appended claims rather than by the specific embodiments that have been represented by way of example.

Claims

1. Formula Ie, 【Chemical 1】 or a pharmaceutically acceptable salt thereof, During the ceremony: R １ is -CH ３ or -CF3; R ４ teeth, 【Chemistry 2】 is; R ５ is methyl, ethyl, propyl, i-propyl, butyl, s-butyl, t-butyl, straight-chain or branched pentyl, or straight-chain or branched hexyl; k is 1; n is 0; p is 0; r is 1; and The compound, or a pharmaceutically acceptable salt thereof, wherein t is 1.

2. The table below: 【Table 1】 2. The compound of claim 1 selected from:

3. A pharmaceutical composition comprising a compound of claim 1 or 2 and a pharmaceutically acceptable adjuvant, carrier, or vehicle.

4. A pharmaceutical composition for inhibiting TLR7 / 8 (or a mutant thereof) activity in a patient or in a biological sample, said pharmaceutical composition comprising a compound according to claim 1 or 2 or a physiologically acceptable salt thereof.

5. 10. A pharmaceutical composition for the treatment of a TLR7 / 8 mediated disorder in a patient in need thereof, comprising a compound according to claim 1 or 2 or a physiologically acceptable salt thereof.

6. 6. The pharmaceutical composition of claim 5, wherein the disorder is selected from rheumatoid arthritis, psoriatic arthritis, osteoarthritis, lupus, systemic lupus erythematosus, lupus nephritis, ankylosing spondylitis, osteoporosis, systemic sclerosis, multiple sclerosis, psoriasis, type I diabetes, type II diabetes, inflammatory bowel disease (Crohn's disease and ulcerative colitis), hyper-IgDemia and periodic fever syndrome, cryopyrin-associated periodic syndrome, Schnitzler's syndrome, systemic juvenile idiopathic arthritis, adult-onset Still's disease, gout, pseudogout, SAPHO syndrome, Castleman's disease, sepsis, stroke, atherosclerosis, celiac disease, DIRA (IL-1 receptor antagonist deficiency), Alzheimer's disease, Parkinson's disease, Sjogren's disease, polymyositis, dermatomyositis, and cancer.

7. 10. A pharmaceutical composition for treating cancer in a subject, comprising a compound according to claim 1 or 2 or a physiologically acceptable salt thereof.

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