Substituted quinoline as an NLRP3 modulator

Abstract

The present invention provides a compound of formula (I), (II), or (III). All variable groups herein are as defined herein. These compounds are modifiers of NLRP3, which can be used as medicines for the treatment of proliferative disorders, such as cancer, in subjects (e.g., humans).

Description

Technology Field

[0001] Cross-reference regarding related applications

[0002] This application claims the benefit of priority of U.S. provisional application No. 62 / 791,967 filed January 14, 2019; the full text of which is incorporated herein by reference.

[0003] Technology field

[0004] The present disclosure features chemical substances (e.g., compounds or pharmaceutically acceptable salts of compounds, and / or co-crystals, and / or drug combinations) that modulate NLRP3 (e.g., agonistic or partially agonistic) which are useful for treating said condition, disease, or disorder in which an increase in NLRP3 signaling can correct a deficiency in innate immune activity that contributes to the pathological state and / or symptoms and / or progression and / or treatment refractory state of said condition, disease, or disorder (e.g., cancer with low T-cell infiltration) of said condition, disease, or disorder in a subject (e.g., human). The present disclosure also features compositions as well as other methods of use and methods of preparation thereof. Background Technology

[0005] Nucleotide-binding oligomerized domain-like receptors ("NLRs") include a family of intracellular receptors that detect pathogen-associated molecular patterns ("PAMPs") and endogenous molecules (see, e.g., Ting, JPY et al., "The NLR gene family: a standard nomenclature," Immunity, 28(3):285-287, (2008)).

[0006] NLRP represents a subfamily of NLRs comprising proteins such as NLRP1, NLRP3, NLRP4, NLRP6, NLRP7, and NLRP12 that contain a pyrin domain. NLRP is believed to be associated with the formation of a multiprotein complex named the inflammasome (see, for example, [Chaput, C. et al., "NOD-like receptors in lung diseases," Frontiers in Immunology, 4: article 393, (2013)]). These complexes typically comprise one or two NLR proteins, an apoptosis-associated speck-like CARD domain-containing (ASC) adapter molecule, and pro-caspase-1 F (see, e.g., Bauernfeind, F and Hornung, V. "Of inflammasomes and pathogens-sensing of microbes by the inflammasome," EMBO Molecular Medicine, 5(6):814-826, (2013)).

[0007] These inflammasomes are formed by the NLRP3 scaffold, ASC adapter, and pro-caspase-1 (see, e.g., [Hirota, JA, et al., "The airway epithelium nucleotide-binding domain and leucine-rich repeat protein 3 inflammasome is activated by urban particulate matter," Journal of Allergy and Clinical Immunology, 129(4):1116.e6-1125.e6, (2012)]), and their expression is thought to be induced by inflammatory cytokines and TLR agonists in bone marrow cells and human bronchial epithelial cells (ibid.). The NLRP3 inflammasome is thought to mediate the caspase-1-dependent conversion of pro-IL-1β and pro-IL-18 to IL-1β and IL-18. Furthermore, IL-1β and IL-18 have potential in the treatment of various types of cancer (e.g., see the literature [Chen, LC. et al., EMBO Mol Med., 4(12):1276-1293 (2012) and Tse, B. WC. et al., PLoS One, 6(9):e24241 (2011)]). IL-18 has been shown to overcome resistance to checkpoint inhibitors in animal tumor models of colon cancer (e.g., see the literature [Ma, Z. et al., Clin. Cancer Res. Jan 11. (2016) DOI: 10.1158 / 1078-0432.CCR-15-1655]).

[0008] The present invention relates to a compound of formula (I), (II), or (III):

[0009]

[0010] Herein, all variable units are as defined in the following text.

[0011] The pharmaceutically permissible salts, stereoisomers, tautomers, and solvates of the compound of formula (I) or (II) are also within the scope of the present invention.

[0012] The present invention also relates to a pharmaceutical composition comprising one or more compounds of the present invention. The present invention also relates to a method for treating cancer using one or more compounds of the present invention.

[0013] The present invention also provides a method for preparing a compound of formula (I) or (II) or a salt, stereoisomer, tautomer, and solvate thereof that is pharmaceutically acceptable thereto, and an intermediate.

[0014] The compound of the present invention can be used in therapy.

[0015] The compound of the present invention can be used in the manufacture of medicines for the treatment of cancer.

[0016] The compounds of the present invention may be used alone, in combination with other compounds of the present invention, or in combination with one or more other agents.

[0017] Other features and advantages of the present invention will become apparent from the following detailed description and claims. Specific details for implementing the invention

[0018] Compound of the present invention

[0019] In a first aspect, the present invention provides, in particular, a compound of formula (I), (II) or (III) or its stereoisomers, tautomers, or pharmaceutically acceptable salts:

[0020]

[0021] Here

[0022] W is independently -YR 6 , -QYR 6 , -QR 6a , and R 6b Selected from;

[0023] Q is independently NR 5 , CHR 5 Selected from , O, and S;

[0024] Y independently C 1-10 Alkylene, C 2-10 Alkenylene, and C 2-10 Selected from alkynylenes, each of which has 0 to 4 R e Replaced with and / or one of the following optionally interposed in each of these:

[0025] (i) O;

[0026] (ii) N(R f );

[0027] (iii) 0 to 4 R g C substituted with 3-6 Cycloalkylene;

[0028] (iv) 0 to 4 R d Phenylene substituted with;

[0029] (v) comprising 5 to 10 ring atoms, wherein 1 to 4 ring atoms are each independently N, N(R f Selected from ), O, and S, and 0 to 4 R d Heteroarylene substituted with; or

[0030] (vi) comprising 3 to 10 ring atoms, wherein 1 to 3 ring atoms are each independently N, N(R f ), O and S(O) 1-2 Selected from, 0 to 4 R g Heterocycloalkylene substituted with;

[0031] R 1 and R 3 In each case, H, halogen, cyano, and C independently 1-4 Alkyl, C 1-4 Haloalkyl, C 1-4 Alkoxy, and C 1-4 Selected from haloalkoxy;

[0032] R 1ais independently halogen, cyano, C 1-4 Alkyl, C 1-4 Haloalkyl, C 1-4 Alkoxy, and C 1-4 Selected from haloalkoxy;

[0033] R 2 is a heteroaryl comprising five ring atoms independently, wherein 1 to 4 ring atoms are each independently selected from N, NH, O, and S, and the heteroaryl has 0 to 3 R d Replaced with;

[0034] R 4a is independently halogen, cyano, C 1-4 Alkyl, C 1-4 Haloalkyl, C 1-4 Alkoxy, C 1-4 Haloalkoxy, N(C 1-4 -(C containing alkyl)2, and 5 ring atoms 0-3 Selected from alkylene)-heteroaryl, wherein 1 to 4 ring atoms are each independently N, NH, N(C 1-4 Selected from alkyl), O, and S, and the heteroaryl has 0 to 3 R d Replaced with;

[0035] R 4 is independently H or R 4a And;

[0036] R 5 is independently H or C 1-4 It is alkyl;

[0037] R 6 -OR independently a , C 1-4 Haloalkoxy, -C(O)R a , -CO2R a , -SO 1-2 (R h ), -CONR i R j , cyano and R 6a Selected from;

[0038] R6a is independently 0 to 4 R d phenyl substituted with; a heteroaryl comprising 5 to 10 ring atoms (wherein 1 to 4 ring atoms are each independently N, N(R f Selected from ), O, and S, and the heteroaryl has 0 to 4 R d Replaced with); 0 to 4 R g C substituted with 3-10 cycloalkyl; and heterocyclyl comprising 3 to 10 ring atoms (wherein 1 to 3 ring atoms are each independently N, N(R f ), O and S(O) 1-2 Selected from, the heterocyclil is 0 to 4 R g Selected from (replaced as);

[0039] R 6b C independently 1-6 Alkoxy, C 1-4 Haloalkoxy, -C(O)R a , -CO2R a , -SO 1-2 (R h ), -CONR i R j , 0 to 4 R d phenyl substituted with; a heteroaryl comprising 5 to 10 ring atoms (wherein 1 to 4 ring atoms are each independently N, N(R f Selected from ), O, and S, and the heteroaryl has 0 to 4 R d Replaced with); 0 to 4 R g C substituted with 3-10 Cycloalkyl; and Heterocyclil selected from (wherein the heterocyclil is 0 to 2 R g Selected from (replaced as);

[0040] R 7 is independently H, halogen, cyano, C 1-4 Alkyl, C 1-4 Haloalkyl, C 1-4Alkoxy, C 1-4 Haloalkoxy, or C 3-6 It is cycloalkyl;

[0041] R 7a is independently a halogen, C 1-4 alkyl or C 3-6 It is cycloalkyl;

[0042] R a is independently H; 0 to 2 R e C substituted with 1-8 Alkyl; -(C 0-3 Alkylene)-C 3-10 Cycloalkyl (wherein the cycloalkyl is 0 to 4 R g Substituted with); containing 3 to 10 ring atoms -(C 0-3 Alkylene)-heterocyclyl (wherein 1 to 3 ring atoms are each independently N(R f Selected from ), O, and S, and the heterocyclil has 0 to 4 R g Replaced with); -(C 0-3 Alkylene)-(C 6-10 aryl) (where aryl is 0 to 4 R d Substituted with); and -(C containing 5 to 10 ring atoms 0-3 Alkylene)-heteroaryl (wherein 1 to 4 ring atoms are each independently N, N(R f Selected from ), O, and S, and the heteroaryl has 0 to 4 R d Selected from (replaced as);

[0043] R b and R c R independently in each case a or -C(O)R a And;

[0044] R d is independently halogen, OH, cyano, C 1-4 Alkoxy, C 1-4 Alkoxy, C 1-4 Haloalkyl, C 1-4 Haloalkoxy, -C(O)O(C 1-4alkyl), NH2, N(C 1-4 Alkyl)2, -C(O)NH2, -C(O)N(C 1-4 Alkyl)2, C 2-6 Alkenyl, C 2-6 Alkynyl, and 0 to 2 R e C substituted with 1-6 Selected from alkyls;

[0045] R e is independently F, OH, cyano, C 1-4 Alkoxy, C 1-4 Haloalkyl, C 1-4 Haloalkoxy, and 0 to 1 R n C substituted with 1-4 Selected from alkyls;

[0046] R f is independently H, C 1-4 Alkyl, -C(O)(C 1-4 alkyl), and -C(O)O(C 1-4 Selected from alkyl;

[0047] R g is independently iodine or R d And;

[0048] R h C independently 1-6 Alkyl, C 1-4 Haloalkyl, -(C 0-3 -(C)-phenyl, comprising 5 to 6 ring atoms, and alkylene)-phenyl, and -(C 0-3 Selected from alkylene)-heteroaryl, wherein 1-4 ring atoms are each independently N, N(R f Selected from ), O, and S;

[0049] R i and R j H or R independently in each case h or; or R i and R j Each forms a ring comprising 5 to 6 ring atoms together with the nitrogen atom to which it is attached, wherein the ring is (a) each of which is H and R m3 to 5 cyclic carbon atoms substituted with 1 to 2 substituents independently selected from; and (b) (R i and R j In addition to the nitrogen atoms attached to it) each independently N(R f It comprises 0 to 2 cyclic heteroatoms selected from ), O, and S;

[0050] R m is independently iodine or R e And;

[0051] R n is independently OH, CONH2, and C 1-4 It is selected from alkoxy.

[0052] In the second aspect, within the category of the first aspect,

[0053] R 1 , R 3 and R 7 In each case, H, halogen, cyano, and C independently 1-4 Alkyl, C 1-4 Haloalkyl, C 1-4 Alkoxy, and C 1-4 Selected from haloalkoxy;

[0054] R 7a is independently a halogen.

[0055] In the third aspect, within the category of the first aspect or the second aspect,

[0056] Q are independently NH, N(C 1-4 Selected from alkyl), CH2, and O;

[0057] Y independently C 1-10 Alkylene, C 2-6 Alkenylene, and C 2-6 Selected from alkynylenes, each of which has 0 to 4 R e Replaced with and / or one of the following optionally interposed in each of these:

[0058] (i) O;

[0059] (ii) N(Rf );

[0060] (iii) 0 to 4 R g C substituted with 3-6 Cycloalkylene;

[0061] (iv) 0 to 4 R d Phenylene substituted with;

[0062] (v) comprising 5 to 6 ring atoms, wherein 1 to 4 ring atoms are each independently N, N(R f Selected from ), O, and S, and 0 to 4 R d Heteroarylene substituted with; or

[0063] (vi) comprising 3 to 7 ring atoms, wherein 1 to 3 ring atoms are each independently N, N(R f ), O and S(O) 1-2 Selected from, 0 to 4 R g Heterocycloalkylene substituted with;

[0064] R 2 is independently a 5-membered heteroaryl comprising one to two ring atoms selected from N, NH, O, and S, each independently;

[0065] R 4a is independently halogen, cyano, C 1-4 Alkyl, C 1-4 Haloalkyl, C 1-4 Alkoxy, C 1-4 Haloalkoxy, N(C 1-4 Selected from 5-membered heteroaryls comprising alkyl)2, and each independently 1 to 2 ring atoms selected from N, NH, O, and S;

[0066] R 4 is independently H or R 4a And;

[0067] R a is independently H, 0 to 2 R e C substituted with 1-6Selected from alkyl, and benzyl;

[0068] R h C independently 1-6 It is alkyl or benzyl;

[0069] R i and R j H or R independently in each case h am.

[0070] In another aspect, within any one of the first to third aspects,

[0071] R 2 is independently selected from pyrazolyl, thienyl, and isothiazolyl;

[0072] R 4a is independently halogen, cyano, C 1-4 Alkyl, C 1-4 Haloalkyl, C 1-4 Alkoxy, C 1-4 Haloalkoxy, N(C 1-4 alkyl)2, and heteroaryl selected from pyrazolyl, thienyl, and isothiazolyl;

[0073] R 4 is independently H or R 4a am.

[0074] In the fourth aspect, within any one of the first to third aspects,

[0075] W is independently R 6 , -YR 6 , -OR 6a , -NH-R 6a , -OYR 6 , -NH-YR 6 , Selected from, where each ring moiety is 0 to 2 R g Replaced with;

[0076] Y independently C 1-8 Alkylene or C 2-6 They are alkynylenes, each of which has 0 to 4 R e Replaced with;

[0077] R 1 is independently H or a halogen;

[0078] R 3 , R 4 and R 7 In each case, H, halogen, and C independently 1-4 Selected from alkyls;

[0079] R 1a is independently a halogen;

[0080] R 4a is independently a halogen or C 1-4 It is alkyl;

[0081] R 7a is independently F or Cl;

[0082] R 2 is independently in each case And;

[0083] R 6 is independently H, OH, C 1-6 Alkoxy, N(C 1-4 Alkyl)2, C 1-6 Haloalkyl, cyano, and R 6a Selected from;

[0084] R 6a is independently 0 to 3 R d phenyl substituted with; a heteroaryl comprising 5 to 10 ring atoms (wherein 1 to 4 ring atoms are each independently N, N(R f Selected from ), O, and S, and the heteroaryl has 0 to 3 R d Replaced with); 0 to 3 R g C substituted with 3-6 Cycloalkyl; heterocyclyl comprising 3 to 8 ring atoms (wherein 1 to 3 ring atoms are each independently N, N(R f ), O and S(O) 1-2 Selected from, the heterocyclil is 0 to 3 R g Replaced with); and Selected from;

[0085] R d is independently halogen, cyano, OH, CH2OH, C 1-4 Alkoxy, C 1-4 Haloalkyl, N(C 1-4 alkyl)2, and 0 to 2 C 1-4 C substituted with an alkoxy 1-4 Selected from alkyls;

[0086] R e is independently F, OH, -(CH2) 1-4 OH, -CH2CONH2, and 1 C 1-4 C substituted with an alkoxy 1-4 Selected from alkyls;

[0087] R f is independently H or C 1-4 It is alkyl;

[0088] R g is independently iodine or R d am.

[0089] In the fifth aspect, within any one of the first to fourth aspects,

[0090] W is independently R 6 , -YR 6 , -NH-R 6a , and -NH-YR 6 Selected from;

[0091] Y independently has 0 to 1 R e C substituted with 1-6 It is an alkylene;

[0092] R 1 , R 3 and R 7 is independently selected from H, F, and Cl in each case;

[0093] R 4 is independently selected from H, F, and CH3 in each case;

[0094] R 1a and R 4ais independently selected from F, Cl, and CH3 in each case;

[0095] R 6 is independently H, OH, C 1-6 Alkoxy, CN, C 1-6 haloalkyl, and R 6a Selected from;

[0096] R 6a is independently 0 to 1 R g Pyrazolyl substituted with , 0 to 2 R g C substituted with 3-6 Cycloalkyl; and Selected from;

[0097] R g is independently halogen, OH, CH2OH, C 1-4 Alkoxy, C 1-4 haloalkyl, and 0 to 2 C 1-4 C substituted with an alkoxy 1-4 Selected from alkyls;

[0098] R e is independently F or OH.

[0099] In the sixth aspect, within any one of the first to fifth aspects,

[0100] W is independently -NH-R 6a or -NH-YR 6 And;

[0101] Y independently C 1-4 It is an alkylene;

[0102] R 1 is independently H or F;

[0103] R 3 and R 7 is independently selected from H, F, and Cl in each case;

[0104] R 4 is independently selected from H, F, and CH3 in each case;

[0105] R 1ais independently F or Cl;

[0106] R 4a is independently selected from F, Cl, and CH3;

[0107] R 6 is independently OH, OCH3 and R 6a Selected from;

[0108] R 6a is independently pyrazolyl, cyclopentyl substituted with OH; and Selected from.

[0109] In another aspect, the present invention provides a compound selected from the exemplified Examples 1 to 16 or its stereoisomers, tautomers, or pharmaceutically acceptable salts.

[0110] In another aspect, the present invention provides a list of any subset of compounds from the examples exemplified within the category of any of the above aspects, or a compound selected from a single compound.

[0111] In some embodiments, R 2 is independently pyrazolyl, thienyl, or isothiazolyl. In other embodiments, R 2 is pyrazolil. In another embodiment, R 2 is thienyl. In another embodiment, R 2 is isothiazolyl.

[0112] A person skilled in the art will recognize that some of the chemical structures described herein may be represented on paper by one or more different resonance forms; or that one or more other tautomeric forms may exist, even if a person skilled in the art recognizes that such tautomeric forms represent only a very small portion of the sample of these compound(s). Such compounds are clearly considered within the scope of this disclosure, but such resonance forms or tautomeric forms are not explicitly shown herein.

[0113] Other aspects and embodiments of the present invention

[0114] In one aspect, the invention features a method for modulating NLRP3 activity (e.g., agonist, partial agonist, antagonist thereof) comprising contacting NLRP3 with a chemical substance described herein (e.g., a compound or a pharmaceutically acceptable salt thereof or a composition containing it as generally or specifically described herein). In a preferred embodiment, the method for modulating NLRP3 activity is an agonist or a partial agonist. In a specific embodiment, the method for modulating NLRP3 activity is an agonist. In a specific embodiment, the method for modulating NLRP3 activity is a partial agonist. The method comprises an in vitro method of contacting a chemical substance with a sample comprising, for example, one or more types of cells (e.g., THP-1 cells) containing NLRP3. The method may also include an in vivo method of administering a chemical substance to a subject (e.g., human) having said disease, in which, for example, an increase in NLRP3 signaling can correct a deficiency in innate immune activity that contributes to the pathological state and / or symptoms and / or progression of said disease (e.g., cancer; e.g., refractory cancer).

[0115] In some embodiments, the compounds of the present invention are useful for treating said condition, disease, or disorder in which a reduction in NLRP3 activity in a subject (e.g., human) (e.g., a condition, disease, or disorder associated with inhibition or impaired NLRP3 signaling) contributes to the pathological state and / or symptoms and / or progression of said condition, disease, or disorder (e.g., cancer).

[0116] Cancer is referred to as refractory when it does not respond to (or is resistant to) cancer treatment. Refractory cancer is also known as resistant cancer.

[0117] In another aspect, a method for treating cancer is characterized by administering an effective amount of the chemical substance described herein (e.g., a compound generally or specifically described herein, or a pharmaceutically acceptable salt thereof, or a composition containing it) to a subject requiring treatment for cancer. In some embodiments, the cancer may be refractory cancer.

[0118] In an additional aspect, a method for treating the said disease, which can correct a deficiency in innate immune activity in which an increase in NLRP3 signaling contributes to the pathological state and / or symptoms and / or progression of the disease, is characterized by comprising administering an effective amount of the chemical substance described herein (e.g., a compound generally or specifically described herein or a pharmaceutically acceptable salt thereof or a composition containing it) to a subject requiring such treatment.

[0119] In another aspect, the treatment method is characterized by administering an effective amount of the chemical substance described herein (e.g., a compound or a pharmaceutically acceptable salt thereof or a composition containing the same) to a subject having a disease in which an increase in NLRP3 signaling can correct a deficiency in innate immune activity that contributes to the pathological state and / or symptoms and / or progression of the disease.

[0120] In an additional aspect, a treatment method comprises administering to a subject a chemical substance described herein (e.g., a compound or a pharmaceutically acceptable salt thereof or a composition containing the same) which is generally or specifically described herein, said chemical substance is administered in an amount effective for treating said disease,

[0121] An embodiment may include one or more of the following features.

[0122] The chemical substance may be administered in combination with one or more additional cancer therapies (e.g., surgery, radiation therapy, chemotherapy, toxin therapy, immunotherapy, cryotherapy, or gene therapy, or combinations thereof; e.g., cancer therapies including the administration of one or more (e.g., two, three, four, five, six, or more) additional anticancer agents). Non-limiting examples of additional anticancer agents (chemotherapy agents) include alkylating agents (e.g., cisplatin, carboplatin, mechloretamine, cyclophosphamide, chlorambucil, ifosfamide, and / or oxaliplatin); anmetanosides (e.g., azathioprine and / or mercaptopurines); terpenoids (e.g., vinca alkaloids and / or taxanes; e.g., vincristine, vinblastine, vinorelbine and / or vindecin, taxol, paclitaxel, and / or docetaxel); Topoisomerases (e.g., type I topoisomerase and / or type II topoisomerase; e.g., camptothecin, e.g. irinotecan and / or topotecan; amsacrin, etoposide, etoposide phosphate and / or tenifoside); cytotoxic antibiotics (e.g., actinomycin, anthracyclines, doxorubicin, daunorubicin, valueusin, idarubicin, epirubicin, bleomycin, plicamycin and / or mitomycin); hormones (e.g., luteinizing hormone-releasing hormone agonists; e.g., leuprolidine, goserelin, triptorelin, histrelin, bicalutamide, flutamide and / or nilutamide); Antibodies (e.g., apsiximab, adalimumab, alemtuzumab, atlizumab, basiliximab, belimumab, bevacizumab, brentuximab vedotin, canakinumab, cetuximab, sertolizumab pegol, daclizumab, denosumab, eculizumab, epalizumab, gemtuzumab, golimumab, golimumab, ibritumomab tiuxetane, infliximab, ipilimumab, muromonab-CD3, natalizumab, ofatumumab, omalizumab, palivizumab, panitumumab, ranibizumab, rituximab, tocilizumab, tocitumomab and / or trastuzumab); anti-angiogenic agents; cytokines; anticoagulants; growth inhibitors; antiparasitics;and CTLA-4, PD-1, PD-L1, PD-1-PD-L1, PD-1-PD-L2, T cell immunoglobulin and mucin 3 (TIM3 or HAVCR2), galectin 9-TIM3, phosphatidylserine-TIM3, lymphocyte activation gene 3 protein (LAG3), MHC class II-LAG3, 4-1BB-4-1BB ligand, OX40-OX40 ligand, GITR, GITR ligand-GITR, CD27, CD70-CD27, TNFRSF25, TNFRSF25-TL1A, CD40L, CD40-CD40 ligand, HVEM-LIGHT-LTA, HVEM, HVEM-BTLA, HVEM-CD160, HVEM-LIGHT, HVEM-BTLA-CD160, CD80, CD80- Immunological checkpoint receptors and other immunomodulators selected from the group consisting of PDL-1, PDL2-CD80, CD244, CD48-CD244, CD244, ICOS, ICOS-ICOS ligand, butyrophyllin including B7-H3, B7-H4, VISTA, TMIGD2, HHLA2-TMIGD2, and BTNL2, members of the Siglec family, TIGIT and PVR families, KIR, ILT and LIR, NKG2D and NKG2A, MICA and MICB, CD244, CD28, CD86-CD28, CD86-CTLA, CD80-CD28, phosphatidylserine, TIM3, phosphatidylserine-TIM3, SIRPA-CD47, VEGF, neurophyllin, CD160, CD30, and CD155 (e.g., CTLA-4 or PD1 or PD-L1), e.g. It is selected from immune checkpoint inhibitors targeting interleukin-2 (IL-2), indoleamine 2,3-deoxygenase (IDO), IL-10, transforming growth factor-β (TGFβ), CD39, CD73 adenosine-CD39-CD73, and CXCR4-CXCL12.;

[0123] The subject may have cancer; for example, the subject has received, is receiving, or will receive one or more types of cancer therapy.

[0124] Non-limiting examples of cancer include acute myeloid leukemia, adrenocortical carcinoma, Kaposi's sarcoma, lymphoma, anal cancer, appendiceal cancer, teratoid / rhabdomyoplastic tumor, basal cell carcinoma, cholangiocarcinoma, bladder cancer, bone cancer, brain cancer, breast cancer, bronchial tumor, carcinoid tumor, heart tumor, cervical cancer, chordoma, chronic lymphocytic leukemia, chronic myeloproliferative neoplasm, colon cancer, colorectal cancer, craniopharyngioma, endometrial cancer, ependymoma, esophageal cancer, sensory neuroblastoma, Ewing sarcoma, eye cancer, fallopian tube cancer, gallbladder cancer, gastric carcinoid tumor, gastric stromal tumor, germ cell tumor, hairy cell leukemia, head and neck cancer, heart cancer, liver cancer, hypopharyngeal cancer, pancreatic cancer, kidney cancer, laryngeal cancer, chronic myeloid leukemia, lip and oral cancer, lung cancer, melanoma, Merkel cell carcinoma, mesothelioma, stomatal cancer, oral cancer, It includes osteosarcoma, ovarian cancer, penile cancer, pharyngeal cancer, prostate cancer, rectal cancer, salivary gland cancer, skin cancer, small intestine cancer, soft tissue sarcoma, testicular cancer, throat cancer, thyroid cancer, urethral cancer, uterine cancer, vaginal cancer, and vulvar cancer.

[0125] In another embodiment, the mammal is identified as having cancer or an infectious disease. Representative infectious diseases are, without limitation, Acinobacter ( Acinobacter ) infection, actinomycosis, African sleeping sickness, acquired immunodeficiency syndrome, amoebiasis, anaplasmosis, anthrax, Arcanobacterium haemoliticum( Arcanobacterium haemolyticum ) infection, Argentine hemorrhagic fever, ascariasis, aspergillosis, astrovirus infection, babesiosis, Bacillus cereus( Bacillus cereus ) infection, bacterial pneumonia, bacterial vaginosis, Bacteroides( Bacteroides ) infection, balantidium, vilerus scaris( Baylisascaris ) infection, BK virus infection, melanosis, Blastocystis hominis( Blastocystic hominis ) infection, blastomycosis, Bolivian hemorrhagic fever, botulism, Brazilian hemorrhagic fever, brucellosis, bubonic plague, Burgholderia( Burkholderi ) infection, Buruli ulcer, calicivirus( Calicivirus ) infection, Campylobacteritis, Candidiasis, feline abrasion, cellulitis, Chagas disease, chancroid, chickenpox, Chikungunya, Chlamydia, Chlamydophila pneumoniae( Chlamydophila pneumoniae ) infection, cholera, chromatophoresis, liver fluke infection, Clostridium difficile( Clostridium difficile ) infection, coccidioidomycosis, Colorado tick fever, common cold, Creutzfeldt-Jakob disease, Crimean-Congo hemorrhagic fever, cryptococcosis, cryptosporidiosis, cutaneous larval migration, cytosporosis, cysticercosis, cytomegalovirus infection, dengue fever, desmodesmus( Desmodesmus ) infection, dientamebiasis, diphtheria, tapeworm infection, guinea worm infection, Ebola hemorrhagic fever, echinococcus, ehrlichiosis, pinworm infection, enterococcus( Enterococcus ) infection, enterovirus( Enterovirus ) infection, epidemic typhus, erythematous infection, sudden rash, hypertrophic trematode disease, epilepsy, fatal familial insomnia, filariasis, Clostridium myonecrosis ( Clostridium myonecrosis Food poisoning caused by ), free-living amoeba infection, Fusobacterium ( Fusobacterium ) infection, gas gangrene, geotricholism, Gerstmann-Straussler-Scheinker syndrome, giardiasis, maggotism, gnatism, gonorrhea, inguinal granuloma, Group A Streptococcus infection, Group B Streptococcus infection, to Haemophilus influenzae ( Haemophilus influenzae ) infection, hand, foot, and mouth disease, Hantavirus pulmonary syndrome, Heartland virus disease, Helicobacter pylori( Heliobacter pylori ) Infection, Hemolytic-Urelytic Syndrome, Hemorrhagic Fever with Renal Syndrome, Hepatitis A, Hepatitis B, Hepatitis C, Hepatitis D, Hepatitis E, Herpes Simplex, Histoplasmosis, Hookworm Infection, Human Bocavirus Infection, Human Ewingie's Ehrlichiosis, Human Granulocytic Anaplasmosis, Human Metapneumovirus Infection, Human Mononucleic Ehrlichiosis, Human Papillomavirus Infection, Human Parainfluenza Virus Infection, Dwarf Tachycystosis, Epstein-Barr Infectious Mononucleosis, Influenza, Sporosis, Kawasaki Disease, Keratitis, Kingella Kingae ( Kingella kingae ) Infection, Kuru, Lassa fever, Legionellosis, Pontiac fever, Leishmaniasis, Leptospirosis, Listeriosis, Lyme disease, Lymphatic filariasis, Lymphocytic choriomeningitis, Malaria, Marburg hemorrhagic fever, Measles, Middle East Respiratory Syndrome, Melioidosis, Meningitis, Meningococcal disease, Yokogawa fluke infection, Microsporidiasis, Infectious molluscum contagiosum, Bean, Mumps, Typhus fever, Mycoplasma pneumoniae( mycoplasma pneumonia ), fungal species, mycosis, neonatal conjunctivitis, variant Creutzfeldt-Jakob disease, nocardiosis, onchocerciasis, paracoccidioidomycosis, paroxysmal pulmonary emphysema, pasteurellosis, lice capitis, lice corporis, lice pubis, pelvic inflammatory disease, pertussis, bubonic plague, pneumonia, poliomyelitis, Prevotella( Prevotella ) Infection, Primary Amoebic Meningoencephalitis, Progressive Multifocal Leukoencephalopathy, Psittacosis, Q fever, Rabies, Relapsing fever, Respiratory syncytial virus infection, Rhinosporidiosis, Rhinovirus infection, Rickettsia infection, Rickettsia pox, Rift Valley fever, Rocky Mountain spotted fever, Rotavirus infection, Rubella, Salmonellosis, Severe Acute Respiratory Syndrome, Scabies, Schistosomiasis, Sepsis, Shigelosis, Herpes zoster, Smallpox, Sporotrichum, Staphylococcus food poisoning, Staphylococcus infection, Strongyloides, Subacute sclerosing panencephalitis, Syphilis, Tapeworm infection, Tetanus, Ringworm, Tinea capitis, Tinea corporis, Tinea cruris, Tinea manus, Ringworm melanoma, Tinea pedis, Tinyfoot, Onychomycosis, Ringworm versicolor, Toxocariasis, Trachoma, Toxoplasmosis, trichinosis, trichomoniasis, whipworm infection, tuberculosis, tularemia, typhoid fever, Ureaplasma urealyticum ( Ureaplasma urealyticum ) infection, Valley fever, Venezuelan hemorrhagic fever, viral pneumonia, West Nile fever, white sarcoma, Yersinia pseudotuberculousis( Yersinia psuedotuberculosis ) Includes infection, yersiniasis, yellow fever, and zygomycosis.

[0126] Chemical substances can be administered into the tumor.

[0127] Chemical substances may be administered systemically (including, but not limited to, orally, subcutaneously, intramuscularly, and intravenously).

[0128] The method may additionally include verifying the object.

[0129] Other embodiments include those described in the detailed description and / or claims.

[0130] definition

[0131] To facilitate understanding of the disclosure presented herein, a number of additional terms are defined below. In general, the nomenclature and laboratory procedures used herein in organic chemistry, medicinal chemistry, and pharmacology are widely known and commonly used in the relevant technical field. Unless otherwise defined, all technical and scientific terms used herein generally have the same meaning as commonly understood by a person skilled in the art to which this disclosure belongs.

[0132] Unless otherwise specifically stated herein, references to the singular may also include the plural. For example, the singular form may refer to one or more than one.

[0133] Unless otherwise indicated, any heteroatom with an unsatisfied valence is assumed to have enough hydrogen atoms to satisfy the valence.

[0134] For clarity and in accordance with standard regulations in the relevant technical field, symbols It is used in chemical formulas and tables to present the bonds that are the attachment points of moiety or substituents to the core / nucleus of the structure.

[0135] Additionally, for clarity, when a substituent has a dash (-) between two letters or symbols, this is used to indicate the attachment point for the substituent. For example, -OCH3 is attached through the oxygen atom.

[0136] The term "NLRP3" as used herein is intended to include, but not limited to, nucleic acids, polynucleotides, oligonucleotides, sense and antisense polynucleotide strands, complementary sequences, peptides, polypeptides, proteins, homologous and / or heterologous NLRP3 molecules, isoforms, precursors, mutants, variants, derivatives, splice variants, alleles, different species, and their active fragments.

[0137] "Agonists" of NLRP3 include compounds that directly bind to or modify NLRP3 at the protein level so as to increase the activity of NLRP3, for example, through activation, stabilization, altered distribution, etc.

[0138] Certain compounds described herein that act on NLRP3 to a lesser extent than full NLRP3 agonists may function as agonists as well as antagonists during testing. Because these compounds block the maximum effect of the NLRP3 interaction, they antagonize the activation of NLRP3 by full NLRP3 agonists. However, these compounds also themselves activate some NLRP3 activity, typically to a lesser extent than a corresponding amount of full NLRP3 agonist. These compounds may be referred to as "partial agonists of NLRP3."

[0139] In some embodiments, the compound described herein is an agonist of NLRP3 (e.g., a full agonist). In other embodiments, the compound described herein is a partial agonist of NLRP3.

[0140] Generally, receptors exist in active (Ra) and inactive (Ri) stereotypes. Certain compounds affecting the receptor can alter the ratio of Ra to Ri (Ra / Ri). For example, full agonists increase the Ra / Ri ratio and can induce a "maximum" saturation effect. Partial agonists, upon binding to the receptor, provide a lower response than that elicited by full agonists (e.g., endogenous agonists). Therefore, the Ra / Ri for partial agonists is lower than that for full agonists. However, the potency of partial agonists can be greater or less than that of full agonists.

[0141] The term "acceptable" with respect to the formulations, compositions, or components used herein means that they do not have a sustained harmful effect on the overall health of the subject to be treated.

[0142] "API" refers to the active pharmaceutical ingredient.

[0143] As used herein, the terms “effective dose” or “therapeutic effective dose” refer to a sufficient amount of a chemical substance to be administered (e.g., a compound exhibiting activity as a mitochondrial decoupling agent or its pharmaceutically acceptable salts and / or hydrates and / or cocrystals; e.g., a compound such as niclosamide or its pharmaceutically acceptable salts and / or hydrates and / or cocrystals; e.g., a compound such as a niclosamide analog or its pharmaceutically acceptable salts and / or hydrates and / or cocrystals) to alleviate, to some extent, one or more of the symptoms of the disease or condition to be treated. The result includes a reduction and / or alleviation of the signs, symptoms, or causes of the disease, or any other intended alteration of the biological system. For example, an “effective dose” for therapeutic use is an amount of a composition containing a compound as disclosed herein that is necessary to provide a clinically significant reduction in the symptoms of the disease. In any individual case, an appropriate “effective” dose is determined using any suitable technique, such as a dose-escalation study.

[0144] The term “excipient” or “pharmaceutical-acceptable excipient” means a pharmaceutically acceptable substance, composition, or vehicle, such as a liquid or solid filler, diluent, carrier, solvent, or encapsulating material. In one embodiment, each component is “pharmaceutical-acceptable” in terms of compatibility with other components of the pharmaceutical formulation and suitability for use in contact with tissues or organs of humans and animals without excessive toxicity, irritation, allergic reaction, immunogenicity, or other problems or complications corresponding to a reasonable benefit / risk ratio. For example, the literature [Remington: The Science and Practice of Pharmacy, 22nd Edition, Pharmaceutical Press, London, UK (2012); Handbook of Pharmaceutical Excipients, 6th ed.; Rowe et al., Eds.; The Pharmaceutical Press and the American Pharmaceutical Association: (2009); Handbook of Pharmaceutical Additives, 3rd ed.; Ash and Ash Eds.; Gower Publishing Company: (2007); Pharmaceutical Preformulation and Formulation, 2nd ed.; See Gibson Ed.; CRC Press LLC: Boca Raton, FL, (2009)].

[0145] The term “pharmaceutical acceptable salt” refers to a preparation of a compound that does not cause significant irritation to an organism to which said salt is administered and does not eliminate the biological activity and properties of the compound. In certain cases, the pharmaceutically acceptable salt is obtained by reacting the compound described herein with an acid, such as hydrochloric acid, hydrobromide, sulfuric acid, nitric acid, phosphoric acid, methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid, salicylic acid, etc. In some cases, the pharmaceutically acceptable salt is obtained by reacting the compound having an acidic group described herein with a base to form a salt, such as an ammonium salt, an alkali metal salt, such as a sodium or potassium salt, an alkaline earth metal salt, such as a calcium or magnesium salt, an organic base, such as a salt of dicyclohexylamine, N-methyl-D-glucarmine, or tris(hydroxymethyl)methylamine, and a salt with an amino acid, such as arginine or lysine, etc., or by other previously determined methods. Pharmacologically acceptable salts are not specifically limited as long as they can be used in medicine. Examples of salts formed by the compounds described herein with bases include salts with inorganic bases such as sodium, potassium, magnesium, calcium, and aluminum; salts with organic bases such as methylamine, ethylamine, and ethanolamine; salts with basic amino acids such as lysine and ornithine; and ammonium salts. The salts may be acid addition salts, which are specifically exemplified by inorganic acids such as hydrochloric acid, hydrobromide, hydroiodide, sulfuric acid, nitric acid, and phosphoric acid; organic acids such as formic acid, acetic acid, propionic acid, oxalic acid, malonic acid, succinic acid, fumaric acid, maleic acid, lactic acid, malic acid, tartaric acid, citric acid, methanesulfonic acid, and ethanesulfonic acid; and acid addition salts with acidic amino acids such as aspartic acid and glutamic acid.

[0146] The term “pharmaceutical composition” refers to a mixture of the compound described herein and other chemical components (collectively referred to herein as “excipients”), such as carriers, stabilizers, diluents, dispersants, suspending agents, and / or thickeners. A pharmaceutical composition facilitates the administration of the compound to an organism. Numerous techniques for administering the compound exist in the relevant art, including but not limited to rectal, oral, intravenous, aerosol, parenteral, ocular, pulmonary, and topical administration.

[0147] The term "subject" refers to an animal including, but not limited to, primates (e.g., humans), monkeys, cattle, pigs, sheep, goats, horses, dogs, cats, rabbits, rats, or mice. The terms "subject" and "patient" are used interchangeably herein with respect to, for example, mammalian subjects, such as humans.

[0148] In relation to treating a disease or disorder, the terms “treat,” “treating,” and “treatment” are intended to include the alleviation or elimination of one or more of the disorder, disease, or condition, or symptoms associated with the disorder, disease, or condition; or the slowing of the progression, spread, or exacerbation of the disease, disorder, or condition, or one or more of its symptoms. “Treatment of cancer” is the following effect: (1) inhibition to some extent of tumor growth, including (i) slowing and (ii) complete growth arrest; (2) reduction in the number of tumor cells; (3) maintenance of tumor size; (4) reduction in tumor size; (5) inhibition of tumor cell infiltration into peripheral organs, including (i) reduction, (ii) slowing, or (iii) complete prevention; (6) inhibition of metastasis, including (i) reduction, (ii) slowing, or (iii) complete prevention; (7) (i) maintenance of tumor size, (ii) reduction of tumor size, (iii) slowing of tumor growth, (iv) enhancement of an anti-tumor immune response capable of reducing, slowing, or preventing invasion, and / or (8) reduction to some degree of the severity or number of one or more symptoms associated with the disorder.

[0149] The term "halo" or "halogen" refers to fluoro (F), chloro (Cl), bromo (Br), or iodo (I).

[0150] The term "alkyl" refers to a hydrocarbon chain that may be straight or branched, containing the indicated number of carbon atoms. For example, C 1-10 The above group indicates that it may have 1 to 10 carbon atoms (including upper and lower limits). Non-limiting examples include methyl, ethyl, iso-propyl, tert-butyl, and n-hexyl.

[0151] The term "alkylene" refers to branched or unbranched divalent alkyl (e.g., -CH2-).

[0152] The term "haloalkyl" refers to an alkyl group in which one or more hydrogen atoms are replaced by independently selected halos.

[0153] The term "alkoxy" refers to -O-alkyl radicals (e.g., -OCH3).

[0154] The term "haloalkoxy" refers to an -O-haloalkyl group as defined above, having the indicated number of carbon atoms and attached via oxygen crosslinking. For example, "C 1-6 "Haloalkoxy" is intended to include C1, C2, C3, C4, C5, and C6 haloalkoxy groups. Examples of haloalkoxy include, but are not limited to, trifluoromethoxy, 2,2,2-trifluoroethoxy, and pentafluoroethoxy.

[0155] The term "alkenyl" refers to a hydrocarbon chain that may be straight or branched and has one or more carbon-carbon double bonds. An alkenyl moiety contains the indicated number of carbon atoms. For example, C 2-6 The above indicates that the group may have 2 to 6 carbon atoms (including upper and lower limits) within it.

[0156] The term "alkynyl" refers to a hydrocarbon chain that may be straight or branched and has one or more carbon-carbon triple bonds. An alkynyl moiety contains the indicated number of carbon atoms. For example, C 2-6 The above indicates that the group may have 2 to 6 carbon atoms (including upper and lower limits) within it.

[0157] The term "aromatic" generally refers to a ring containing a resonance-stabilized cyclic arrangement of 4n + 2 pi electrons where n is an integer (e.g., 1 or 2). Aromatic moiety includes aryl and heteroaryl groups. The term "non-aromatic" describes any moiety that does not fall under the definition of "aromatic."

[0158] The term "aryl" refers to a 6-carbon monocyclic, 10-carbon bicyclic, or 14-carbon tricyclic aromatic ring system, wherein 0, 1, 2, 3, or 4 atoms of each ring may be substituted by substituents, wherein the ring constituting the monocyclic radical is aromatic, and wherein at least one of the fused rings constituting the bicyclic or tricyclic radical is aromatic, e.g., tetrahydronaphthyl. Examples of aryl groups also include phenyl, naphthyl, etc.

[0159] As used herein, the term "cycloalkyl" comprises a saturated cyclic hydrocarbon group having 3 to 10 carbons, preferably 3 to 8 carbons, more preferably 3 to 6 carbons, wherein the cycloalkyl group may optionally be substituted. Preferred cycloalkyl groups include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl, cycloheptyl, and cyclooctyl. As used herein, the term "cycloalkylene" refers to a divalent cycloalkyl.

[0160] The term "heteroaryl" refers to an aromatic 5-8-membered monocyclic, 8-12-membered cyclic, or 11-14-membered tricyclic ring system having 1-3 heteroatoms in the case of monocyclic, 1-6 heteroatoms in the case of cyclic, or 1-9 heteroatoms in the case of tricyclic, wherein the heteroatoms are selected from O, N, or S (e.g., carbon atoms, and having 1-3, 1-6, or 1-9 heteroatoms of N, O, or S in the case of monocyclic, cyclic, or tricyclic, respectively), wherein 0, 1, 2, 3, or 4 atoms of each ring may be substituted by substituents, wherein the ring constituting the monocyclic radical is aromatic, and wherein at least one of the fused rings constituting the cyclic or tricyclic radical is aromatic (however, the ring containing heteroatoms (It is not necessary), for example, tetrahydroisoquinolinyl. Examples of heteroaryl groups also include pyridyl, furyl or furanyl, imidazolyl, benzimidazolyl, pyrimidinyl, thiophenyl or thienyl, quinolinyl, indolyl, thiazolyl, etc.

[0161] The term "heterocyclyl" refers to a non-aromatic 5-8-membered monocyclic, 8-12-membered cyclic, or 11-14-membered tricyclic ring system having 1-3 heteroatoms in the case of monocyclic, 1-6 heteroatoms in the case of cyclic, or 1-9 heteroatoms in the case of tricyclic, wherein the heteroatoms are selected from O, N, or S (e.g., carbon atoms, and having 1-3, 1-6, or 1-9 heteroatoms of N, O, or S in the case of monocyclic, cyclic, or tricyclic, respectively), wherein 0, 1, 2, or 3 atoms of each ring may be substituted by substituents. Examples of heterocyclyl groups include piperazinyl, pyrrolidinyl, dioxanyl, morpholinyl, tetrahydrofuranyl, etc. The term "heterocycloalkylene" refers to divalent heterocyclyl.

[0162] Additionally, the atoms constituting the compounds of embodiments of the present invention are intended to include all isotopic forms of such atoms. The isotopes used herein include these atoms having the same atomic number but different mass numbers. As a general example and without limitation, isotopes of hydrogen include tritium and deuterium, and isotopes of carbon are 13 C and 14 Includes C.

[0163] Details of one or more embodiments of the present invention are set forth in the accompanying drawings and the following description. Other features and advantages of the present invention will be apparent from the description, drawings, and claims.

[0164] The present disclosure features, for example, chemical substances (e.g., compounds or pharmaceutically acceptable salts and / or hydrates and / or co-crystals and / or drug combinations) that modulate NLRP3 (e.g., agonistic or partially agonistic) which are useful for treating said condition, disease, or disorder (e.g., a condition, disease, or disorder associated with an insufficient immune response) in which an increase in NLRP3 signaling can correct a deficiency in innate immune activity that contributes to the pathological state and / or symptoms and / or progression of said condition, disease, or disorder (e.g., cancer) in a subject (e.g., human). The present disclosure also features compositions as well as other methods of use and methods of preparation thereof.

[0165] Pharmaceutical composition and administration

[0166] In some embodiments, a chemical substance (e.g., a compound that modifies NLRP3 (e.g., an agonist or partial agonist) or a pharmaceutically acceptable salt and / or hydrate thereof and / or cocrystal and / or drug combination) is administered as a pharmaceutical composition comprising said chemical substance and one or more pharmaceutically acceptable excipients, and optionally one or more additional therapeutic agents as described herein.

[0167] In some embodiments, the pharmaceutical composition comprises a compound of the present invention or a salt thereof and one or more pharmaceutically acceptable excipients. In certain embodiments, the pharmaceutical composition comprises a compound of the present invention or a pharmaceutically acceptable salt thereof and one or more pharmaceutically acceptable excipients. In certain embodiments, the pharmaceutical composition comprises a therapeutically effective amount of a compound of the present invention or a pharmaceutically acceptable salt thereof and one or more pharmaceutically acceptable excipients.

[0168] In some embodiments, the chemical substance may be administered in combination with one or more conventional pharmaceutical excipients. Pharmaceutically acceptable excipients include, but are not limited to, ion exchangers, alumina, aluminum stearate, lecithin, self-emulsifying drug delivery systems (SEDDS) such as d-α-tocopherol polyethylene glycol 1000 succinate, surfactants used in pharmaceutical dosage forms such as Tween, poloxamer or other similar polymer delivery matrices, serum proteins such as human serum albumin, buffering materials such as phosphate, tris, glycine, sorbic acid, potassium sorbate, mixtures of partial glycerides 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 materials, polyethylene glycol, sodium carboxymethyl cellulose, polyacrylates, waxes, polyethylene-polyoxypropylene-block polymers, and wool fats. Cyclodextrins, such as α-, β-, and γ-cyclodextrins, or chemically modified derivatives, such as hydroxyalkylcyclodextrins including 2- and 3-hydroxypropyl-β-cyclodextrins, or other solubilized derivatives may also be used to enhance the delivery of the compounds described herein. Dosage forms or compositions may be prepared containing the chemical as described herein in a range of 0.005% to 100% and the remainder consisting of non-toxic excipients. The compositions under consideration may contain the chemical provided herein in an amount of 0.001% to 100%, 0.1% to 95% in some embodiments, 75% to 85% in other embodiments, and 20% to 80% in additional embodiments.The actual method of preparing this dosage form will be known or obvious to a person skilled in the art; for example, see the literature [Remington: The Science and Practice of Pharmacy, 22nd Edition (Pharmaceutical Press, London, UK. 2012)].

[0169] Route of administration and composition components

[0170] In some embodiments, the chemical substance or its pharmaceutical composition described herein may be administered to a subject in need of it by any acceptable route of administration. Permitted routes of administration include, but are not limited to: buccal, cutaneous, intracervical, sinusoidal, tracheal, transcutaneous, epidural, interstitial, intraperitoneal, intra-arterial, intrabronchial, bursal, intracerebral, cisternous, coronary, intradermal, intraductal, duodenal, intradural, intraepidermal, intraesophageal, gastric, gingival, ileal, lymphatic, medullary, meningeal, intramuscular, ovarian, intraperitoneal, prostate, lung, sinus or periorbital sinus, spinal cord, synovial, testicular, spinal canal, tubular, tumor, uterine, intravascular, intravenous, nasal, nasogastric tube, oral, parenteral, percutaneous, percutaneous, rectal, respiratory (inhalation), subcutaneous, sublingual, submucosal, local, percutaneous, transmucosal, trans-tracheal, ureteral, urethra, and vagina. In certain embodiments, the preferred route of administration is parenteral (e.g., intratumoral). In certain embodiments, the preferred route of administration is systemic.

[0171] The composition may be formulated for parenteral administration and may be formulated for injection via, for example, intravenous, intramuscular, subcutaneous, or even intraperitoneal routes. Typically, such composition may be prepared as an injectable preparation as a liquid solution or suspension; a solid form suitable for use in preparing a solution or suspension upon the addition of liquid prior to injection may also be prepared; and such preparations may also be emulsified. Methods for preparing such preparations will be known to those skilled in the art in light of the present disclosure.

[0172] Pharmaceutical forms suitable for injectable use include sterile aqueous solutions or dispersions; preparations containing sesame oil, peanut oil, or aqueous propylene glycol; and sterile powders for the immediate preparation of sterile injectable solutions or dispersions. In all cases, the form must be sterile and fluid enough to be easily injected. It must also be stable under manufacturing and storage conditions and preserved against the action of microorganisms, such as bacteria and fungi.

[0173] The carrier may also be a solvent or dispersion medium containing, for example, water, ethanol, polyols (e.g., glycerol, propylene glycol, and liquid polyethylene glycol, etc.), suitable mixtures thereof, and vegetable oil. Suitable fluidity can be maintained, for example, by using a coating such as lecithin, by maintaining the particle size required for the dispersion, and by using a surfactant. Prevention of microbial action can be achieved by various antibacterial and antifungal agents, for example, parabens, chlorobutanol, phenol, sorbic acid, thimerosal, etc. In many cases, it will be desirable to include an isotonic agent, for example, sugar or sodium chloride. Sustained absorption of the injectable composition can be achieved by using an absorption-delaying agent, for example, aluminum monostearate and gelatin, in the composition.

[0174] A sterile injectable solution is prepared by incorporating a required amount of an active compound into a suitable solvent along with, if necessary, various other components listed above, and then filtering and sterilizing. Generally, a dispersion is prepared by incorporating various sterile active ingredients into a sterile vehicle containing a basic dispersion medium and other necessary components from those listed above. In the case of a sterile powder for the preparation of a sterile injectable solution, a preferred manufacturing method is vacuum-drying and freeze-drying techniques that produce a powder of the active ingredient plus any additional desired component from its previously sterile-filtered solution.

[0175] Intratumoral injection is discussed, for example, in the literature [Lammers, et al., "Effect of Intratumoral Injection on the Biodistribution and the Therapeutic Potential of HPMA Copolymer-Based Drug Delivery Systems" Neoplasia. 10:788-795 (2006)].

[0176] Pharmacologically acceptable excipients usable in rectal compositions as gels, creams, enemas, or rectal suppositories are, without limitation, cocoa butter glycerides, synthetic polymers such as polyvinylpyrrolidone, PEG (e.g., PEG ointment), glycerin, glycerinated gelatin, hydrogenated vegetable oils, poloxamer, Vaseline (a mixture of polyethylene glycol of various molecular weights and fatty acid esters of polyethylene glycol), anhydrous lanolin, shark liver oil, sodium saccharinate, menthol, sweet almond oil, sorbitol, sodium benzoate, anoxidized SBN, vanilla essential oil, parabens in aerosols and phenoxyethanol, sodium methyl p-oxybenzoate, sodium propyl p-oxybenzoate, diethylamine, carbomer, carbopol, methyloxybenzoate, macrogol cetostearyl ether, cocoyl caprylocaprate, and isopropyl It includes any one or more of alcohol, propylene glycol, liquid paraffin, xanthan gum, carboxy-metabisulfite, sodium edetate, sodium benzoate, potassium metabisulfite, grapefruit seed extract, methylsulfonyl methane (MSM), lactic acid, glycine, vitamins, such as vitamins A and E and potassium acetate.

[0177] In certain embodiments, suppositories may be prepared by mixing the chemical described herein with a suitable non-irritating excipient or carrier, such as cocoa butter, polyethylene glycol, or suppository wax, which is solid at ambient temperature but liquid at body temperature and thus melts rectum to release the active compound. In other embodiments, the composition for rectal administration is in the form of an enema.

[0178] In other embodiments, the compound or pharmaceutical composition thereof described herein is suitable for local delivery to the digestive tract or GI tract by oral administration (e.g., in the form of a solid or liquid dose).

[0179] Solid dosage forms for oral administration include capsules, tablets, pills, powders, and granules. In these solid dosage forms, the chemical comprises one or more pharmaceutically acceptable excipients, e.g., sodium citrate or dicalcium phosphate, and / or a) fillers or extenders, e.g., starch, lactose, sucrose, glucose, mannitol, and silicic acid; b) binders, e.g., carboxymethylcellulose, alginate, gelatin, polyvinylpyrrolidinone, sucrose, and acacia; c) humectants, e.g., glycerol; d) disintegrants, e.g., agar-agar, calcium carbonate, potato or tapioca starch, alginic acid, certain silicates, and sodium carbonate; e) dissolution retardants, e.g., paraffin; f) absorption promoters, e.g., quaternary ammonium compounds; g) wetting agents, e.g., cetyl alcohol and glycerol monostearate; h) absorbents, e.g., kaolin and bentonite clay; and i) lubricants, e.g., talc. It is mixed with calcium stearate, magnesium stearate, solid polyethylene glycol, sodium lauryl sulfate, and mixtures thereof. In the case of capsules, tablets, and pills, the dosage form may also include a buffer. Similar types of solid compositions may also be used as fillers in soft and hard-filled gelatin capsules using excipients such as lactose or lactose, as well as high molecular weight polyethylene glycol, etc.

[0180] In one embodiment, the composition will take the form of a unit dosage form, e.g., a pill or a tablet, and thus the composition may contain, together with the chemical provided herein, a diluent, e.g., lactose, sucrose, dical phosphate, etc.; a lubricant, e.g., magnesium stearate, etc.; and a binder, e.g., starch, acacia gum, polyvinylpyrrolidine, gelatin, cellulose, cellulose derivatives, etc. In another solid dosage form, a powder, marum, solution, or suspension (e.g., among propylene carbonate, vegetable oil, PEG, poloxamer 124, or triglycerides) is encapsulated within a capsule (a gelatin or cellulose-based capsule). Unit dosage forms in which one or more of the chemical substances or additional active agents provided herein are physically distinct; e.g., a capsule (or tablet within a capsule) having granules of each drug; a two-layer tablet; a two-compartment gel cap, etc. are also considered. Enteric-coated or delayed-release oral dosage forms are also considered.

[0181] Other physiologically acceptable compounds include wetting agents, emulsifiers, dispersants, or preservatives that are particularly useful for preventing the growth or action of microorganisms. Various preservatives are widely known and include, for example, phenol and ascorbic acid.

[0182] In certain embodiments, the excipients are sterile and do not contain generally undesirable substances. These compositions may be sterilized by conventional and widely known sterilization techniques. For various oral dosage forms of excipients, such as tablets and capsules, sterility is not required. USP / NF standards are typically sufficient.

[0183] In certain embodiments, the solid oral administration form may further comprise one or more components that cause the composition to have a tendency to chemically and / or structurally deliver the chemical to the stomach or lower GI; e.g., the ascending colon and / or transverse colon and / or distal colon and / or small intestine. Exemplary formulation techniques are described, for example, in the literature [Filipski, KJ, et al., Current Topics in Medicinal Chemistry, 2013, 13, 776-802], the full text of which is incorporated herein by reference.

[0184] Examples include upper-GI targeting technologies, such as accordion pills (Intec Pharma), floating capsules, and materials capable of adhering to mucosal walls.

[0185] Other examples include lower-GI targeting technologies. For targeting various regions of the intestinal tract, various enteric / pH-responsive coatings and excipients are available. These materials are typically polymers designed to dissolve or corrode within a specific pH range selected based on the GI region of the desired drug release. These materials also function to protect acid-unstable drugs from gastric juice or to limit exposure when the active ingredient may be irritating to the upper GI (e.g., hydroxypropyl methylcellulose phthalate series, Coateric (polyvinyl acetate phthalate), cellulose acetate phthalate, hydroxypropyl methylcellulose acetate succinate, Eudragit series (methacrylate-methyl methacrylate copolymer), and Marcoat). Other technologies include local flora-responsive dosage forms in the GI tube, pressure-controlled colon delivery capsules, and Pulsincap.

[0186] The ocular composition may, without limitation, include any one or more of viscogen (e.g., carboxymethylcellulose, glycerin, polyvinylpyrrolidone, polyethylene glycol); stabilizers (e.g., Pluronic (triblock copolymer), cyclodextrin); preservatives (e.g., benzalkonium chloride, ETDA, SofZia (boric acid, propylene glycol, sorbitol, and zinc chloride; Alcon Laboratories, Inc.), and Purite (stabilized oxychloro complex; Allergan, Inc.).

[0187] Topical compositions may include ointments and creams. Ointments are typically semi-solid formulations based on petrolatum or other petroleum derivatives. Creams containing selected active agents are typically viscous liquids or semi-solid emulsions that are often oil-in-water or water-in-oil. Cream bases are typically water-washable and contain an oil phase, an emulsifier, and an aqueous phase. The oil phase, sometimes also referred to as the "internal" phase, generally consists of petrolatum and fatty alcohols such as cetyl or stearyl alcohols; the aqueous phase typically exceeds the oil phase in volume, though not necessarily, and generally contains a humectant. The emulsifier in the cream formulation is generally a nonionic, anionic, cationic, or amphoteric surfactant. Like other carriers or vehicles, the ointment base must be inert, stable, non-irritating, and non-sensitizing.

[0188] In any of the above embodiments, the pharmaceutical composition described herein may comprise one or more of lipids, multilayer vesicles cross-linked within a bilayer, biodegradable poly(D,L-lactic acid-co-glycolic acid) [PLGA]-based or polyanhydride-based nanoparticles or microparticles, and a nanoporous particle-supported lipid bilayer.

[0189] Dosage

[0190] The dosage may vary depending on the patient's requirements, the severity of the condition to be treated, and the specific compound used. The determination of the appropriate dosage for a specific situation may be made by a person skilled in the art of medical technology. The total daily dosage may be divided and administered in multiple portions throughout the day or by means of providing continuous delivery.

[0191] In some embodiments, the compounds described herein are in an amount of about 0.001 mg / kg to about 500 mg / kg (e.g., about 0.001 mg / kg to about 200 mg / kg; about 0.01 mg / kg to about 200 mg / kg; about 0.01 mg / kg to about 150 mg / kg; about 0.01 mg / kg to about 100 mg / kg; about 0.01 mg / kg to about 50 mg / kg; about 0.01 mg / kg to about 10 mg / kg; about 0.01 mg / kg to about 5 mg / kg; about 0.01 mg / kg to about 1 mg / kg; about 0.01 mg / kg to about 0.5 mg / kg; about 0.01 mg / kg to about 0.1 mg / kg; about 0.1 mg / kg to about 200 mg / kg; about 0.1 mg / kg to about It is administered at doses of 150 mg / kg; about 0.1 mg / kg to about 100 mg / kg; about 0.1 mg / kg to about 50 mg / kg; about 0.1 mg / kg to about 10 mg / kg; about 0.1 mg / kg to about 5 mg / kg; about 0.1 mg / kg to about 1 mg / kg; about 0.1 mg / kg to about 0.5 mg / kg).

[0192] therapy

[0193] The above dosage may be administered on a daily basis (e.g., as a single dose or as two or more divided doses), or on a non-daily basis (e.g., every other day, every 2 days, every 3 days, once a week, twice a week, once every 2 weeks, once a month).

[0194] In some embodiments, the duration of administration of the compound described herein is 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, 8 days, 9 days, 10 days, 11 days, 12 days, 13 days, 14 days, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 7 weeks, 8 weeks, 9 weeks, 10 weeks, 11 weeks, 12 weeks, 4 months, 5 months, 6 months, 7 months, 8 months, 9 months, 10 months, 11 months, 12 months, or longer than that. In additional embodiments, the period during which administration is discontinued is 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, 8 days, 9 days, 10 days, 11 days, 12 days, 13 days, 14 days, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 7 weeks, 8 weeks, 9 weeks, 10 weeks, 11 weeks, 12 weeks, 4 months, 5 months, 6 months, 7 months, 8 months, 9 months, 10 months, 11 months, 12 months, or longer. In one embodiment, the therapeutic compound is administered to an individual for a separate time period following a predetermined time period. In another embodiment, the therapeutic compound is administered for a first period, administration is discontinued for a second period after the first period, administration of the therapeutic compound is then started for a third period, and administration is then discontinued for a fourth period after the third period. In one aspect of such an embodiment, the period of discontinuation of administration following the administration of the therapeutic compound is repeated for a determined or undetermined period of time. In a further embodiment, the administration period is 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, 8 days, 9 days, 10 days, 11 days, 12 days, 13 days, 14 days, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 7 weeks, 8 weeks, 9 weeks, 10 weeks, 11 weeks, 12 weeks, 4 months, 5 months, 6 months, 7 months, 8 months, 9 months, 10 months, 11 months, 12 months, or for a longer period.In additional embodiments, the period during which administration is discontinued is 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, 8 days, 9 days, 10 days, 11 days, 12 days, 13 days, 14 days, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 7 weeks, 8 weeks, 9 weeks, 10 weeks, 11 weeks, 12 weeks, 4 months, 5 months, 6 months, 7 months, 8 months, 9 months, 10 months, 11 months, 12 months, or longer than that.

[0195] Treatment methods

[0196] In some embodiments, a method is provided for treating a subject having said condition, disease, or disorder (e.g., a condition, disease, or disorder associated with an insufficient immune response), wherein an increase in NLRP3 signaling can correct a deficiency in innate immune activity that contributes to the pathological state and / or symptoms and / or progression of said condition, disease, or disorder (e.g., cancer).

[0197] Indications

[0198] In any method described herein, the subject may have cancer. In some examples of any method described herein, the mammal is confirmed to have cancer or is diagnosed to have cancer.

[0199] Non-limiting examples of cancer include acute myeloid leukemia, adrenocortical carcinoma, Kaposi's sarcoma, lymphoma, anal cancer, appendiceal cancer, teratoid / rhabdomyoplastic tumor, basal cell carcinoma, cholangiocarcinoma, bladder cancer, bone cancer, brain cancer, breast cancer, bronchial tumor, carcinoid tumor, heart tumor, cervical cancer, chordoma, chronic lymphocytic leukemia, chronic myeloproliferative neoplasm, colon cancer, colorectal cancer, craniopharyngioma, endometrial cancer, ependymoma, esophageal cancer, sensory neuroblastoma, Ewing sarcoma, eye cancer, fallopian tube cancer, gallbladder cancer, gastric carcinoid tumor, gastric stromal tumor, germ cell tumor, hairy cell leukemia, head and neck cancer, heart cancer, liver cancer, hypopharyngeal cancer, pancreatic cancer, kidney cancer, laryngeal cancer, chronic myeloid leukemia, lip and oral cancer, lung cancer, melanoma, Merkel cell carcinoma, mesothelioma, stomatal cancer, oral cancer, It includes osteosarcoma, ovarian cancer, penile cancer, pharyngeal cancer, prostate cancer, rectal cancer, salivary gland cancer, skin cancer, small intestine cancer, soft tissue sarcoma, testicular cancer, throat cancer, thyroid cancer, urethral cancer, uterine cancer, vaginal cancer, and vulvar cancer.

[0200] In certain embodiments, non-limiting examples of cancer include breast cancer, colon cancer, rectal cancer, colorectal cancer, pancreatic cancer, and prostate cancer.

[0201] Methods for diagnosing a subject as having cancer or confirming a mammal as having cancer are widely known in the relevant art. For example, a medical professional (e.g., a physician, a physician’s assistant, or a technician) may diagnose cancer in a mammal by observing one or more symptoms of cancer in the mammal. Non-limiting examples of symptoms of cancer include fatigue, a lump or thickened area felt under the skin, changes in body weight, jaundice, darkening or redness of the skin, pain that does not heal, changes in existing moles, changes in bowel or bladder habits, persistent cough or difficulty breathing, difficulty swallowing, hoarseness, persistent dyspepsia or postprandial discomfort, unexplained persistent muscle pain or joint pain, unexplained persistent fever or night sweats, and unexplained bleeding or bruising. Methods for diagnosing a subject as having cancer or confirming a subject as having cancer may further include performing one or more diagnostic tests (e.g., performing one or more diagnostic tests on a biopsy or blood sample).

[0202] In some examples of any method described herein, the subject may be a subject having cancer that was unresponsive to previously administered treatment for cancer, a subject diagnosed with cancer, or a subject confirmed to have cancer. Diagnostic tests for diagnosing a subject having cancer or confirming a mammal having cancer are known in the art.

[0203] In some embodiments, a method is provided for treating a subject having said condition, disease, or disorder (e.g., a condition, disease, or disorder associated with an insufficient immune response), wherein an increase in NLRP3 signaling can correct a deficiency in innate immune activity that contributes to the pathological state and / or symptoms and / or progression of said condition, disease, or disorder (e.g., cancer).

[0204] In some embodiments, the present invention provides a method for treating cancer, wherein the cancer may be any cancer that does not elicit an optimal innate immune system response.

[0205] The innate immune system refers to a part of the immune system composed of cells that respond to threats to the organism, such as infections or cancer, in an antigen-nonspecific manner and stimulate the adaptive antigen-specific immune system. Generally, the complete elimination of a threat and long-term sustained defense (i.e., immunity) require the activation of the adaptive antigen-specific immune system, which in turn depends on stimulation by the innate immune system.

[0206] In some embodiments, the present invention provides a method for treating a case in which cancer is selected based on resistance to T-cell checkpoint inhibition, based on failure of response to previous T-cell checkpoint inhibitor therapy regardless of cancer type, or based on cancer types generally resistant to T-cell checkpoint inhibitor therapy, such as hormone receptor-positive breast cancer, microsatellite-stable colon or rectal cancer, pancreatic cancer, and prostate cancer.

[0207] In a specific other embodiment, the present invention provides a method for treating cancer comprising the NLPR3 agonist of the present invention for treating a non-inflammatory tumor having low CD8+ T-cell infiltration to enhance tumor immunogenicity and promote an inflammatory response. For example, the combination may be used to treat a solid tumor based on biopsy results demonstrating low CD8+ T-cell infiltration or low expression of genes produced by CD8+ T-cells.

[0208] Resistance to T-cell checkpoint inhibition refers to cancer progression during therapy or lack of response within 6 months of therapy according to the consensus response criteria for each cancer, e.g., RECIST 1.1 for most solid tumors.

[0209] T-cell infiltration refers to the percentage of T-cells among all nucleated cells determined by immunohistochemistry of a tumor biopsy specimen.

[0210] CD8+ T-cell infiltration refers to the percentage of CD8+ cells among all nucleated cells by immunohistochemistry of a tumor biopsy specimen.

[0211] In addition to immunohistochemistry for quantifying CD8+ T-cells in biopsy specimens, the expression of genes produced by CD8+ T-cells, such as interferon-γ, can be measured by quantifying mRNA using, for example, next-generation sequencing, and CD8+ T-cell infiltration can be indicated. Thresholds for low CD8+ T-cell infiltration and high CD8+ T-cell infiltration by immunohistochemistry of mRNA quantification technology are being developed by various methods, considering the spectrum of CD8+ T-cell infiltration across cancers as well as specific cancers.

[0212] In any method described herein, the subject may have an infectious disease. In some examples of any method described herein, the subject is identified as having an infectious disease or is diagnosed as having an infectious disease. For example, the infectious disease may be caused by bacteria, viruses, fungi, parasites, or mycobacterium.

[0213] Non-limiting examples of infectious diseases include Acinobacter ( Acinobacter ) infection, actinomycosis, African sleeping sickness, acquired immunodeficiency syndrome, amoebiasis, anaplasmosis, anthrax, Arcanobacterium haemoliticum( Arcanobacterium haemolyticum ) infection, Argentine hemorrhagic fever, ascariasis, aspergillosis, astrovirus infection, babesiosis, Bacillus cereus( Bacillus cereus ) infection, bacterial pneumonia, bacterial vaginosis, Bacteroides( Bacteroides ) infection, balantidium, vilerus scaris( Baylisascaris ) infection, BK virus infection, melanosis, Blastocystis hominis( Blastocystic hominis ) infection, blastomycosis, Bolivian hemorrhagic fever, botulism, Brazilian hemorrhagic fever, brucellosis, bubonic plague, Burgholderia( Burkholderi ) infection, Buruli ulcer, calicivirus( Calicivirus) Infection, Campylobacteritis, Candidiasis, Feline Abrasion, Cellulitis, Chagas Disease, Chancroid, Chickenpox, Chikungunya, Chlamydia, Chlamydophila pneumoniae ( Chlamydophila pneumoniae ) infection, cholera, chromatophoresis, liver fluke infection, Clostridium difficile( Clostridium difficile ) infection, coccidioidomycosis, Colorado tick fever, common cold, Creutzfeldt-Jakob disease, Crimean-Congo hemorrhagic fever, cryptococcosis, cryptosporidiosis, cutaneous larval migration, cytosporosis, cysticercosis, cytomegalovirus infection, dengue fever, desmodesmus( Desmodesmus ) infection, dientamebiasis, diphtheria, tapeworm infection, guinea worm infection, Ebola hemorrhagic fever, echinococcus, ehrlichiosis, pinworm infection, enterococcus( Enterococcus ) infection, enterovirus( Enterovirus ) infection, epidemic typhus, erythematous infection, sudden rash, hypertrophic trematode disease, epilepsy, fatal familial insomnia, filariasis, Clostridium myonecrosis ( Clostridium myonecrosis Food poisoning caused by ), free-living amoeba infection, Fusobacterium ( Fusobacterium ) infection, gas gangrene, geotricholism, Gerstmann-Straussler-Scheinker syndrome, giardiasis, maggotism, gnatism, gonorrhea, inguinal granuloma, Group A Streptococcus infection, Group B Streptococcus infection, to Haemophilus influenzae ( Haemophilus influenzae ) infection, hand, foot, and mouth disease, Hantavirus pulmonary syndrome, Heartland virus disease, Helicobacter pylori( Heliobacter pylori ) Infection, Hemolytic-Urelytic Syndrome, Hemorrhagic Fever with Renal Syndrome, Hepatitis A, Hepatitis B, Hepatitis C, Hepatitis D, Hepatitis E, Herpes Simplex, Histoplasmosis, Hookworm Infection, Human Bocavirus Infection, Human Ewingie's Ehrlichiosis, Human Granulocytic Anaplasmosis, Human Metapneumovirus Infection, Human Mononucleic Ehrlichiosis, Human Papillomavirus Infection, Human Parainfluenza Virus Infection, Dwarf Tachycystosis, Epstein-Barr Infectious Mononucleosis, Influenza, Sporosis, Kawasaki Disease, Keratitis, Kingella Kingae ( Kingella kingae ) Infection, Kuru, Lassa fever, Legionellosis, Pontiac fever, Leishmaniasis, Leptospirosis, Listeriosis, Lyme disease, Lymphatic filariasis, Lymphocytic choriomeningitis, Malaria, Marburg hemorrhagic fever, Measles, Middle East Respiratory Syndrome, Melioidosis, Meningitis, Meningococcal disease, Yokogawa fluke infection, Microsporidiasis, Infectious molluscum contagiosum, Bean, Mumps, Typhus fever, Mycoplasma pneumoniae( mycoplasma pneumonia ), fungal species, mycosis, neonatal conjunctivitis, variant Creutzfeldt-Jakob disease, nocardiosis, onchocerciasis, paracoccidioidomycosis, paroxysmal pulmonary emphysema, pasteurellosis, lice capitis, lice corporis, lice pubis, pelvic inflammatory disease, pertussis, bubonic plague, pneumonia, poliomyelitis, Prevotella( Prevotella ) Infection, Primary Amoebic Meningoencephalitis, Progressive Multifocal Leukoencephalopathy, Psittacosis, Q fever, Rabies, Relapsing fever, Respiratory syncytial virus infection, Rhinosporidiosis, Rhinovirus infection, Rickettsia infection, Rickettsia pox, Rift Valley fever, Rocky Mountain spotted fever, Rotavirus infection, Rubella, Salmonellosis, Severe Acute Respiratory Syndrome, Scabies, Schistosomiasis, Sepsis, Shigelosis, Herpes zoster, Smallpox, Sporotrichum, Staphylococcus food poisoning, Staphylococcus infection, Strongyloides, Subacute sclerosing panencephalitis, Syphilis, Tapeworm infection, Tetanus, Ringworm, Tinea capitis, Tinea corporis, Tinea cruris, Tinea manus, Ringworm melanoma, Tinea pedis, Tinyfoot, Onychomycosis, Ringworm versicolor, Toxocariasis, Trachoma, Toxoplasmosis, trichinosis, trichomoniasis, whipworm infection, tuberculosis, tularemia, typhoid fever, Ureaplasma urealyticum ( Ureaplasma urealyticum ) infection, Valley fever, Venezuelan hemorrhagic fever, viral pneumonia, West Nile fever, white sarcoma, Yersinia pseudotuberculousis( Yersinia psuedotuberculosis ) Includes infection, yersiniasis, yellow fever, and zygomycosis.

[0214] Methods for diagnosing or confirming that a subject has an infectious disease are widely known in the relevant art. For example, a medical professional (e.g., a physician, a physician’s assistant, or a technician) may diagnose an infectious disease in a subject by observing one or more symptoms of an infectious disease in the subject. Non-limiting examples of symptoms of an infectious disease include fever, diarrhea, fatigue, and muscle pain. Methods for diagnosing that a mammal has an infectious disease or confirming that a subject has an infectious disease may further include performing one or more diagnostic tests (e.g., performing one or more diagnostic tests on a biopsy or blood sample). Diagnostic tests for diagnosing or confirming that a subject has an infectious disease are known in the relevant art.

[0215] Combination therapy

[0216] The present disclosure considers both monotherapy regimens and combination therapy regimens.

[0217] In some embodiments, the method described herein may further include administering one or more additional therapies (e.g., one or more additional therapeutic agents and / or one or more therapeutic therapies) in combination with the administration of the compound described herein.

[0218] In certain embodiments, the method described herein may further include administering one or more additional cancer therapies.

[0219] One or more additional cancer therapies may include, but are not limited to, surgery, radiation therapy, chemotherapy, toxin therapy, immunotherapy, cryotherapy, cancer vaccines (e.g., HPV vaccine, hepatitis B vaccine, Oncophage, Provenge), and gene therapy, as well as combinations thereof. Immunotherapy may, but is not limited to, adoptive cell therapy, induction of stem cells and / or dendritic cells, blood transfusion, washing, and / or other treatments including, but is not limited to, freezing of the tumor.

[0220] In some embodiments, one or more additional cancer therapies are chemotherapy that may include administering one or more additional chemotherapy agents.

[0221] In certain embodiments, additional cancer therapy includes an immunomodulatory moiety (chemotherapy agent), e.g., an immune checkpoint inhibitor. In certain of these embodiments, the immune checkpoint inhibitor is CTLA-4, PD-1, PD-L1, PD-1-PD-L1, PD-1-PD-L2, T cell immunoglobulin and mucin 3 (TIM3 or HAVCR2), galectin 9-TIM3, phosphatidylserine-TIM3, lymphocyte activation gene 3 protein (LAG3), MHC class II-LAG3, 4-1BB-4-1BB ligand, OX40-OX40 ligand, GITR, GITR ligand-GITR, CD27, CD70-CD27, TNFRSF25, TNFRSF25-TL1A, CD40L, CD40-CD40 ligand, HVEM-LIGHT-LTA, HVEM, HVEM-BTLA, HVEM-CD160, HVEM-LIGHT, HVEM-BTLA-CD160, CD80, CD80-PDL-1, PDL2-CD80, CD244, CD48-CD244, CD244, ICOS, ICOS-ICOS ligand, butyrophyllin including B7-H3, B7-H4, VISTA, TMIGD2, HHLA2-TMIGD2, BTNL2, members of the Siglec family, TIGIT and PVR families, KIR, ILT and LIR, NKG2D and NKG2A, MICA and MICB, CD244, CD28, CD86-CD28, CD86-CTLA, CD80-CD28, phosphatidylserine, TIM3, phosphatidylserine-TIM3, SIRPA-CD47, VEGF, neurophyllin, CD160, CD30, and CD155 (e.g., CTLA-4 or PD1 or It targets immune checkpoint receptors selected from the group consisting of PD-L1 and other immunomodulators, such as interleukin-2 (IL-2), indoleamine 2,3-deoxygenase (IDO), IL-10, transforming growth factor-β (TGFβ), CD39, CD73 adenosine-CD39-CD73, and CXCR4-CXCL12.For example, refer to the literature [Postow, MJ Clin. Oncol. 33, 1 (2015)].

[0222] In a specific embodiment, the immune checkpoint inhibitor targets an immune checkpoint receptor selected from CTLA-4, PD-1, PD-L1, PD-1-PD-L1, and PD-1-PD-L2.

[0223] In certain embodiments, the immune checkpoint inhibitors are nivolumab (also known as "Opdivo"; previously designated as 5C4, BMS-936558, MDX-1106, or ONO-4538), pembrolizumab (also known as "Keytruda", lambrolizumab, and MK-3475; see WO 2008 / 156712), PDR001 (Novartis; see WO 2015 / 112900), MEDI-0680 (AstraZeneca; AMP-514; see WO 2012 / 145493), cemiplimab (REGN-2810) (Regeneron; see WO 2015 / 112800), and JS001 (Taizhou Junshi). Pharma (TAIZHOU JUNSHI PHARMA; see reference [Si-Yang Liu et al., J. Hematol. Oncol. 10:136 (2017)]), BGB-A317 (Beigene; see WO 2015 / 35606 and US 2015 / 0079109), INCSHR1210 (SHR-1210; Jiangsu Hengrui Medicine; WO 2015 / 085847; see reference [Si-Yang Liu et al., J. Hematol. Oncol. 10:136 (2017)]), TSR-042 (ANB011; Tesaro Biopharmaceutical; see WO2014 / 179664), GLS-010 (WBP3055; Wuxi / Harbin Gloria Pharmaceuticals; Literature [Si-Yang Liu et al., J. Hematol. Oncol.[See 10:136 (2017)]), AM-0001 (Armo), STI-1110 (Sorrento Therapeutics; see WO 2014 / 194302), AGEN2034 (Agenus; see WO 2017 / 040790), MGD013 (Macrogenics); IBI308 (Innovent; see WO 2017 / 024465, WO 2017 / 025016, WO 2017 / 132825, WO2017 / 133540); BMS-936559 (formerly 12A4 or MDX-1105; see, e.g., U.S. Patent No. 7,943,743 and WO 2013 / 173223), MPDL3280A (also known as RG7446, atezolizumab, and TeccentriQ; U.S. 8,217,149; see also [Herbst et al. (2013) J Clin Oncol 31(suppl):3000]), durvalumab (IMFINZI; MEDI-4736; AstraZeneca; see WO 2011 / 066389), avelumab (Pfizer; MSB-0010718C; BAVENCIO; see WO 2013 / 079174), STI-1014 (Sorrento; See WO2013 / 181634), CX-072 (Cytomx; see WO2016 / 149201), KN035 (3D Med / Alphamab; see reference [Zhang et al., Cell Discov. 7:3 (March 2017)]), LY3300054 (Eli Lilly Co.; see, e.g., WO 2017 / 034916), CK-301 (Checkpoint Therapeutics; see reference [Gorelik et al., see AACR:Abstract 4606 (Apr 2016)]); Urelumab, PF-05082566, MEDI6469, TRX518, varlilumab, CP-870893, BMS-986016, MGA271, rililumab, IPH2201, emactuzumab, INCB024360, galunicertib, ulocuplumab, BKT140, babituximab, CC-90002, bevacizumab, MNRP1685A, ipilimumab (Yervoy; U.S. Patent No. 6,984,720), MK-1308 (Merck), AGEN-1884 (Agenus Inc.; WO 2016 / 196237), and tremelimumab (formerly ticilimumab, CP-675,206; Selected from AstraZeneca; for example, WO 2000 / 037504 and the literature [Ribas, Update Cancer Ther. 2(3): 133-39 (2007)].

[0224] In certain embodiments, immune checkpoint inhibitors are selected from nivolumab, pembrolizumab, JS001, BGB-A317, INCSHR1210, TSR-042, GLS-010, STI-1110, MGD013, IBI308, BMS-936559, atezolizumab, durvalumab, avelumab, STI-1014, CX-072, KN035, LY3300054, CK-301, urelumab, PF-05082566, MEDI6469, TRX518, varlilumab, BMS-986016, ipilimumab, AGEN-1884, and tremelimumab.

[0225] In these specific embodiments, the immune checkpoint inhibitor is selected from urelumab, PF-05082566, MEDI6469, TRX518, varlilumab, CP-870893, pembrolizumab (PD1), nivolumab (PD1), atezolizumab (formerly MPDL3280A) (PDL1), MEDI4736 (PD-L1), avelumab (PD-L1), PDR001 (PD1), BMS-986016, MGA271, rililumab, IPH2201, emactuzumab, INCB024360, galunisertib, ulokuflumab, BKT140, babituximab, CC-90002, bevacizumab, and MNRP1685A.

[0226] In certain embodiments, immune checkpoint inhibitors are selected from nivolumab, ipilimumab, pembrolizumab, atezolizumab, durvalumab, and avelumab.

[0227] In certain embodiments, the immune checkpoint inhibitor is selected from nivolumab and ipilimumab.

[0228] In certain embodiments, an additional anticancer agent (chemotherapy agent) is a STING agonist. For example, the STING agonist may comprise cyclic dinucleotides, such as cAMP, cGMP, and cGAMP, as well as modified cyclic dinucleotides comprising one or more of the following modification features: a 2'-O / 3'-O linkage, a phosphorothioate linkage, an adenine and / or guanine analogue, and a 2'-OH modification (e.g., -OCH3 or a substitute, e.g., -F or N3). For example, refer to WO 2014 / 189805.

[0229] In certain embodiments, the additional chemotherapeutic agent is an alkylating agent. The alkylating agent is so named because of its ability to alkylate many nucleophilic functional groups in a state of presence within cells, including but not limited to cancer cells. In further embodiments, the alkylating agent includes, but is not limited to, cisplatin, carboplatin, mechlorethamine, cyclophosphamide, chlorambucil, ifosfamide, and / or oxaliplatin. In one embodiment, the alkylating agent may function by impairing cell function by forming covalent bonds with amino, carboxyl, sulfhydryl, and phosphate groups within biologically important molecules, or may act by altering the cell's DNA. In further embodiments, the alkylating agent is synthetic, semi-synthetic, or a derivative.

[0230] In certain embodiments, the additional chemotherapeutic agent is an antimetabolite. The antimetabolite mimics purines or pyrimidines, which are building blocks of DNA, and generally prevents these substances from incorporating into DNA during the "S" phase (of the cell cycle) to halt normal development and division. The antimetabolite may also affect RNA synthesis. In one embodiment, the antimetabolite includes, but is not limited to, azathioprine and / or mercaptopurines. In further embodiments, the antimetabolite is synthetic, semi-synthetic, or a derivative.

[0231] In certain embodiments, additional chemotherapeutic agents are plant alkaloids and / or terpenoids. These alkaloids are derived from plants and block cell division, generally inhibiting microtubule function. In one embodiment, the plant alkaloids and / or terpenoids are vinca alkaloids, podophyllotoxins, and / or taxanes. Vinca alkaloids generally bind to specific sites on tubulin, inhibiting the assembly of tubulin into microtubules, generally during the M phase of the cell cycle. In one embodiment, vinca alkaloids are, without limitation, Madagascar periwinkle ( Madagascar periwinkle ), Cataranthus Roseus( Catharanthus roseus ) (Previously Vinca Rosea( Vinca rosea It is derived from (as known as ). In one embodiment, the vinca alkaloid includes, but is not limited to, vincristine, vinblastine, vinorelbine, and / or vindecin. In one embodiment, the taxane includes, but is not limited to, taxol, paclitaxel, and / or docetaxel. In additional embodiments, the plant alkaloid or terpenoid is synthetic, semi-synthetic, or a derivative. In additional embodiments, the podophyllotoxin is, but is not limited to, etoposide and / or tenifoside. In one embodiment, the taxane is, but is not limited to, docetaxel and / or ortataxel. In one embodiment, the anticancer agent is topoisomerase. Topoisomerase is an essential enzyme that maintains the phase of DNA. Inhibition of type I or type II topoisomerase interferes with both transcription and replication of DNA by overturning proper DNA supercoiling. In additional embodiments, the topoisomerase is, non-limitingly, a type I topoisomerase inhibitor or a type II topoisomerase inhibitor. In one embodiment, the type I topoisomerase inhibitor is, non-limitingly, camptothecin. In another embodiment, camptothecin is, non-limitingly, exatecan, irinotecan, lutetotecan, topotecan, BNP 1350, CKD 602, DB 67 (AR67), and / or ST 1481. In one embodiment, the type II topoisomerase inhibitor is, non-limitingly, epipodophyllotoxin. In additional embodiments, the epipodophyllotoxin is, non-limitingly, amsacrin, etoposide, etoposide phosphate, and / or tenifoside. In additional embodiments, topoisomerase is synthetic, semi-synthetic, or a derivative, and is found in nature, such as, but not limited to, American Mayapple ( American Mayapple ) (Podophilum peltatum( Podophyllum peltatum It contains epipodophyllotoxin, a naturally occurring substance in the roots of )).

[0232] In certain embodiments, the additional chemotherapeutic agent is a stilbenoid. In additional embodiments, the stilbenoid includes, but is not limited to, resveratrol, picetanol, pinosylvin, pterostilbene, alpha-viniferin, ampelopsin A, ampelopsin E, diptoindonesin C, diptoindonesin F, epsilon-viniferin, flexuosol A, gnetin H, hemlesleyanol D, hofeaphenol, trans-diptoindonesin B, astringin, picaide, and diptoindonesin A. In additional embodiments, the stilbenoid is synthetic, semi-synthetic, or a derivative.

[0233] In certain embodiments, the additional chemotherapeutic agent is a cytotoxic antibiotic. In one embodiment, the cytotoxic antibiotic is, but is not limited to, actinomycin, anthracendione, anthracyclin, thalidomide, dichloroacetic acid, nicotinic acid, 2-deoxyglucose, and / or clofazimine. In one embodiment, actinomycin is, but is not limited to, actinomycin D, bacitracin, colistin (polymyxin E), and / or polymyxin B. In another embodiment, the anthracendione is, but is not limited to, mitoxantrone and / or picantrone. In additional embodiments, the anthracyclin is, but is not limited to, bleomycin, doxorubicin (adriamycin), daunorubicin (daunomycin), epirubicin, idarubicin, mitomycin, plicamycin, and / or valubicin. In additional embodiments, the cytotoxic antibiotic is synthetic, semi-synthetic, or a derivative.

[0234] In certain embodiments, additional chemotherapeutic agents include endostatin, angiogenin, angiostatin, chemokine, angioarestin, angiostatin (plasminogen fragment), basement membrane collagen-derived anti-angiogenic factor (tumstatin, canstatin, or arrestin), anti-angiogenic antithrombin III, signal transduction inhibitor, chondrogenic inhibitor (CDI), CD59 complement fragment, fibronectin fragment, gro-beta, heparinase, heparin hexasaccharide fragment, human chorionic gonadotropin (hCG), interferon alpha / beta / gamma, interferon-derived protein (IP-10), interleukin-12, kringle 5 (plasminogen fragment), metalloproteinase inhibitor (TIMP), 2-methoxyestradiol, placental ribonuclease inhibitor, plasminogen activator inhibitor, platelet factor-4 (PF4), prolactin It is selected from 16 kD fragments, proliferin-related protein (PRP), various retinoids, tetrahydrocortisol-S, thrombospondin-1 (TSP-1), transforming growth factor-beta (TGF-β), vasculostatin, vasostatin (caleticulin fragment), etc.

[0235] In certain embodiments, additional chemotherapeutic agents are abiraterone acetate, altretamine, anhydrobinblastine, auristatin, bexarotene, bicalutamide, BMS 184476, 2,3,4,5,6-pentafluoro-N-(3-fluoro-4-methoxyphenyl)benzenesulfonamide, bleomycin, N,N-dimethyl-L-valyl-L-valyl-N-methyl-L-valyl-L-proli-1-L-proline-t-butylamide, kaketin, semadotin, chlorambucil, cyclophosphamide, 3',4'-didehydro-4'-deoxy-8'-norbin-carucoblastine, docetaxol, docetaxel, cyclophosphamide, carboplatin, carmustine, cisplatin, cryptopicin, cyclophosphamide, cytarabine, Dacarbazine (DTIC), Dactinomycin, Daunorubicin, Decitabine, Dolastatin, Doxorubicin (Adriamycin), Etoposide, 5-Fluoruracil, Finasteride, Flutamide, Hydroxyurea and Hydroxyureataxane, Ifosfamide, Riarozole, Ronidamine, Lomustine (CCNU), MDV3100, Mechloretamine (Nitrogen Mustard), Melphalan, Mibobulin Isethionate, Rizoxin, Sertenef, Streptozosin, Mitomycin, Methotrexate, Taxane, Nilutamide, Onapriston, Paclitaxel, Prednimustine, Procarbazine, RPR109881, Stramustine Phosphate, Tamoxifen, Tasonermin, Taxol, Tretinoin, Vinblastine, It is selected from vincristine, vindecin sulfate, and vinflunine.

[0236] In certain embodiments, additional chemotherapeutic agents are platinum, cisplatin, carboplatin, oxaliplatin, mechloretamine, cyclophosphamide, chlorambucil, azathioprine, mercaptopurine, vincristine, vinblastine, vinorelbine, vindecin, etoposide and tenifoside, paclitaxel, docetaxel, irinotecan, topotecan, amsacrine, etoposide, etoposide phosphate, tenifoside, 5-fluorouracil, leucovorin, methotrexate, gemcitabine, taxane, leucovorin, mitomycin C, tegafur-uracil, idarubicin, fludarabine, mitoxantrone, ifosfamide and doxorubicin. Additional agents include inhibitors of mTOR (mammalian target of rapamycin), including but not limited to rapamycin, everolimus, temsirolimus, and deporolimus.

[0237] In another embodiment, additional chemotherapy agents may be selected from those described in U.S. Patent 7,927,613.

[0238] In another embodiment, the method comprises (i) one or more antifungal agents (e.g., bifonazole, butoconazole, clotrimazole, econazole, ketoconazole, luliconazole, miconazole, omoconazole, oxyconazole, sertaconazole, sulfonazole, thioconazole, albaconazole, efinaconazole, epoxyconazole, fluconazole, isavuconazole, itraconazole, posaconazole, propiconazole, labuconazole, terconazole, voriconazole, abafungin, amorolfine, butenafin, naftifine, terbinafine, anidulafungin, caspofungin, micapungin, benzoic acid, ciclopirox, flucytosine, 5-fluorocytosine, griseofulvin, haloprozine, tolnaphtate, undecylenic acid, and (selected from the group of Peru balsam) and (ii) one or more antibiotics (e.g., amikacin, gentamicin, kanamycin, neomycin, netilmycin, tobramycin, paromomycin, streptomycin, spectinomycin, geldanamycin, herbimycin, rifaximin, loracarbev, ertapenem, doripenem, imipenem, cilastatin, meropenem, cefadroxil, cefazolin, cephalotin, cefalothin, cephalexin, cefachlor, cefamandol, cefoxitin, cefprozil, cefuroxim, cefixim, cefdinir, cefditoren, cefoperazone, cefotaxim, cefpodoxim, ceftazidime, ceftibuten, ceftizoxime, ceftriaxone, cefepim, Ceftaroline, fosamil, ceftobiprol, teicoplanin, vancomycin, telavancin, dalbavancin, oritavancin, clindamycin, lincomycin, daptomycin, azithromycin, clarithromycin, dirithromycin, erythromycin, roxithromycin, troleandomycin, telithromycin, spiramycin, aztreonam, furazolidone, nitrofurantoin, linezolid, fosizolid, radezolid, torezolid, amoxicillin, ampicillin, azlocillin, carbenicillin, cloxacillin, dicloxacillin, flucloxacillin, mezlocillin, methicillin, nafcillin, oxacillin, penicillin G, penicillin V, piperacillin, penicillin G, temocillin, ticarcillin, Amoxicillin, Clavulanate, Ampicillin, Sulbactam,Piperacillin, Tazobactam, Ticarcillin, Clavulanate, Bacitracin, Colistin, Polymyxin B, Ciprofloxacin, Enoxacin, Gatifloxacin, Gemifloxacin, Levofloxacin, Lomefloxacin, Moxifloxacin, Naldixic Acid, Norfloxacin, Ofloxacin, Trovafloxacin, Grepafloxacin, Sparfloxacin, Temafloxacin, Mapenide, Sulfacetamide, Sulfadiazine, Sulfadiazine, Sulfadimethoxin, Sulfamethoxazole, Sulfanylimide, Sulfasalazine, Sulfisoxazole, Trimethoprim-Sulfamethoxazole, Sulfonamidochrysoidin, Demeclocycline, Minocyclin, Oxytetracycline, Tetracycline, Clofazimine, Dapsone, It may additionally include administering one or both of the group selected from capreomycin, cycloserine, ethambutol, ethionamide, isoniazid, pyrazinamide, rifampicin, rifabutin, rifapentine, streptomycin, arsphenamine, chloramphenicol, fosfomycin, fusidic acid, metronidazole, mupirocin, platensymycin, quinupristine, dalfopristin, thiamphenicol, tigecycline, tinidazole, trimethoprim, and teixobactin.

[0239] In certain embodiments, the second therapeutic agent or therapy is administered to the subject before contact with or administration of the chemical (e.g., about 1 hour before, or about 6 hours before, or about 12 hours before, or about 24 hours before, or about 48 hours before, or about 1 week before, or about 1 month before).

[0240] In another embodiment, the second therapeutic agent or therapy is administered to the subject at approximately the same time as contacting or administering the chemical. For example, the second therapeutic agent or therapy and the chemical are provided to the subject simultaneously in the same dosage form. As yet another example, the second therapeutic agent or therapy and the chemical are provided to the subject jointly in separate dosage forms.

[0241] In another embodiment, the second therapeutic agent or therapy is administered to the subject after contact with the chemical or after administration thereof (e.g., after about 1 hour, or after about 6 hours, or after about 12 hours, or after about 24 hours, or after about 48 hours, or after about 1 week, or after about 1 month).

[0242] Patient selection

[0243] In some embodiments, the method described herein further comprises the step of identifying a subject (e.g., a patient) requiring such treatment (e.g., by biopsy, endoscopy, or other conventional methods known in the art). In certain embodiments, the NLRP3 protein may function as a biomarker for a specific type of cancer.

[0244] In some embodiments, the chemicals, methods, and compositions described herein may be administered to a specific group of treatment-resistant patients (e.g., patients resistant to checkpoint inhibitors).

[0245] In some embodiments, the compounds of the present invention may be used in therapy. In certain embodiments, the present invention provides a combination formulation of the compound of the present invention, or a pharmaceutically acceptable salt thereof, and additional therapeutic(s) for simultaneous, individual, or sequential use in therapy.

[0246] In some embodiments, the compound of the present invention, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition containing the same may be used as a medicine. In certain embodiments, the compound of the present invention may be used in the manufacture of a medicine for the treatment of cancer. In certain embodiments, the compound of the present invention may be used in the manufacture of a medicine for modulating NLRP3 activity. In certain embodiments, modulation involves acting efferentially on NLRP3.

[0247] Manufacturing method

[0248] As would be recognized by a person skilled in the art, the method for synthesizing the compounds of the formulas of the present invention will be obvious to a person skilled in the art. For example, the compounds described herein may be synthesized, for example, by using one or more of the methods described herein or by using the method described in, for example, US 2015 / 0056224. Synthetic chemical transformations and protecting group methodologies (protection and deprotection) useful for synthesizing the compounds described herein are known in the art, for example, [Larock, RC, Comprehensive Organic Transformations, 2 nd [Includes those described in Edition, Wiley-VCH, New York, NY (1999); Wuts, PGM, Greene's Protective Groups in Organic Synthesis, 5th Edition, Wiley (2014); L. Fieser and M. Fieser, Fieser and Fieser's Reagents for Organic Synthesis, John Wiley and Sons (1994); and L. Paquette, ed., Encyclopedia of Reagents for Organic Synthesis, John Wiley and Sons (1995), and subsequent editions thereof.] The starting materials used in the preparation of the compounds of the present invention are known, prepared by known methods, or are commercially available. A person skilled in the art will also recognize that the conditions and reagents described herein may be interchangeable with alternative equivalents recognized in the relevant art. For example, in many reactions, triethylamine can be interchanged with other bases, such as non-nucleophilic bases (e.g., diisopropylamine, 1,8-diazabicycloundes-7-ene, 2,6-di-tert-butylpyridine, or tetrabutylphosphazene).

[0249] A person of ordinary skill in the art, for example 1 You will recognize various analytical methods that can be used to characterize the compounds described herein, including ¹H NMR, dinuclear NMR, mass spectrometry, liquid chromatography, and infrared spectroscopy. The above list is a subset of characterization methods available to a person skilled in the art and is not intended to be limiting.

[0250] The following abbreviations have the indicated meanings:

[0251] ACN = Acetonitrile

[0252] AcOH = Acetic acid

[0253] CDCl3 = Chloroform-d

[0254] CD3OD = methanol-d4

[0255] CH2Cl2 = Dichloromethane

[0256] CH3ReO3 = Methyltrioxorhenium

[0257] Cs2CO3 = Cesium carbonate

[0258] CuI = copper(I) iodide

[0259] d = double line

[0260] DCM = Dichloromethane

[0261] DIEA = N,N-diethylisopropylamine

[0262] DMF = N,N-dimethylformamide

[0263] DMSO = Dimethyl sulfoxide

[0264] ES = Electrospray ionization

[0265] Et2O = Diethyl ether

[0266] EtOAc = Ethyl acetate

[0267] EtOH = Ethanol

[0268] equiv = equivalent weight

[0269] g = gram

[0270] h = time

[0271] HCl = Hydrogen chloride (usually as a solution)

[0272] H2O = water

[0273] H2O2 = hydrogen peroxide

[0274] HATU = 1-[bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium 3-oxide hexafluorophosphate

[0275] HPLC = High-performance liquid chromatography

[0276] I2 = Iodine

[0277] K2CO3 = potassium carbonate

[0278] K2HPO4 = potassium phosphate, dibasic

[0279] KI = Potassium iodide

[0280] kg = kilogram

[0281] LC / MS = Liquid Chromatography Mass Spectrometer

[0282] LiBH4 = Lithium boroside

[0283] m = multiple lines

[0284] m / z = mass to charge ratio

[0285] M = mole

[0286] m-CPBA = meta-chloroperoxybenzoic acid

[0287] mg = milligram

[0288] MeOH = methanol

[0289] MHz = Megahertz

[0290] mL = milliliters

[0291] mmol = millimole

[0292] min = minutes

[0293] NaHCO3 = sodium bicarbonate

[0294] Na2CO3 = Sodium carbonate

[0295] NaOH = Sodium hydroxide

[0296] Na2SO4 = Sodium sulfate

[0297] NEt3 and TEA = Triethylamine

[0298] NH4OH or NH3H2O ​​= Ammonium hydroxide

[0299] NH4HCO3 = Ammonium bicarbonate

[0300] nm = nanometer

[0301] PdCl2(PPh3)2 = bis(triphenylphosphine)palladium(II) dichloride

[0302] Pd(dppf)Cl2= [1,1'-bis(diphenylphosphino)ferrocene]dichloropalladium(II)

[0303] Pd(dppf)Cl2DCM = [1,1'-Bis(diphenylphosphino)ferrocene]dichloropalladium(II) dichloromethane complex

[0304] Pd(OH)2 = Palladium hydroxide

[0305] PMB = Para-methoxybenzyl

[0306] POCl3 = Phosphorus oxychloride

[0307] ppm = parts per million

[0308] Pt = Platinum

[0309] Pt / C = Carbon phase platinum

[0310] s = single line

[0311] t = triplet

[0312] TFA = Trifluoroacetic acid

[0313] TLC = Thin-layer chromatography

[0314] TsCl = Para-toluenesulfonyl chloride

[0315] ℃ = Celsius temperature

[0316] μmol = micromoles

[0317] The compounds of the present invention can be prepared in a number of ways widely known to those skilled in the art of organic synthesis. The compounds of the present invention can be synthesized using the methods described below, or modifications thereof as recognized by those skilled in the art, in combination with synthesis methods known in the field of synthetic organic chemistry. Preferred methods include, but are not limited to, the methods described below.

[0318] The compounds of the present invention may be prepared using the reactions and techniques described in this section. The reactions are carried out in a solvent suitable for the reagents and materials used and suitable for carrying out the transformation. Furthermore, in the description of the synthesis method described below, all proposed reaction conditions, including the selection of solvent, reaction atmosphere, reaction temperature, duration of experiment, and post-processing procedure, are selected to be standard conditions for the reaction, and it should be understood that this should be readily recognized by a person skilled in the art. It is understood by a person skilled in the art of organic synthesis that functional groups present in various parts of the molecule must be compatible with the proposed reagents and reactions. Such limitations on substituents compatible with the reaction conditions will be readily apparent to a person skilled in the art, and thus alternative methods should be used. This may sometimes require modifying the order of synthesis steps to obtain the desired compounds of the present invention, or making a judgment regarding the selection of one particular process reaction scheme over another. In addition, it will be recognized that another major consideration in planning any synthetic route in this field is the careful selection of a protecting group used to protect the reactive functional groups present in the compounds described in this invention.

[0319] Compounds of formulas (I), (II), or (III) may be prepared by reference to the method exemplified in the following reaction scheme. As indicated therein, the final product is a compound having the same structural formula as formulas (I), (II), or (III). It will be understood that any compound of formulas (I), (II), or (III) may be prepared by the reaction scheme by the appropriate selection of reagents having appropriate substitutions. The solvent, temperature, pressure, and other reaction conditions may be readily selected by a person skilled in the art. The starting materials may be commercially available or readily prepared by a person skilled in the art. The components of the compounds are as defined herein or elsewhere in this specification.

[0320] Chemical formula (I, R 7a The synthesis of compounds (where α is a halogen) can be carried out using the method summarized in reaction schemes 1 and 2.

[0321] Reaction Equation 1

[0322]

[0323] Step 1: The first step of Reaction Scheme 1 begins with a suitably functionalized quinolinol (i). If desired, group R 8 , R 9 , R 10 , and R 11 R, a group present in the compound of chemical formula (I). 1 , R 2 , R 3 , and R 4It may be. Alternatively, one or more of these groups may be groups that can be modified in later stages of synthesis, such as bromo. This quinolinol may be commercially available or synthesized by methods known to those skilled in the art. In Step 1, the alcohol group of compound (i) may be converted to a halogen group or a sulfonate ester, such as chloro, bromo, or tritrilate. If the desired group Z is chloro, this conversion may be carried out by treating compound (i) with a reagent, such as phosphoryl chloride, in a solvent, such as toluene. Alternatively, if the desired group Z is bromo, this conversion may be carried out by treating compound (i) with a reagent, such as tribromide, in a solvent, such as DMF. Alternatively, if the desired group Z is a trilate, this conversion can be carried out by treating compound (i) in a solvent such as dichloromethane with a reagent such as trifluoromethanesulfonyl chloride, a reagent such as 4-dimethylaminopyridine, and a base such as Finnig base.

[0324] Step 2: In Step 2 of Reaction Scheme 1, compound (ii) is treated with a suitable oxidizing agent, such as meta-chloroperoxybenzoic acid, in a solvent such as DCM to convert it into N-oxide (iii).

[0325] Step 3: In Step 3 of Reaction Scheme 1, compound (iii) is treated in a suitable solvent, such as DCM, with a suitable activating reagent, such as tosyl chloride, and an ammonia source, such as ammonium chloride and triethylamine, to convert it into amine (iv).

[0326] Step 4: In Step 4 of Reaction Scheme 1, the halogen Z of compound (iv) is replaced by the group R of compound (v). 12 Converts to. Gi R 12may be the desired group W in the final compound; alternatively, it may be a group that can be converted to group W in later stages of synthesis. A person skilled in the art would know that the means for carrying out such conversion is group R 12 You will be aware that it will vary depending on the properties of Z. For example, if Z is chloro and the target group R 12 In the case where α is an amine, this conversion can be carried out by heating compound (iv) in a solvent such as DMSO or NMP with a suitable amine and a base such as Finnig base to a suitable temperature, e.g., 120°C. Alternatively, Z is chloro and the desired group R is 12 In the case where α is an ether, this conversion can be carried out by heating compound (iv) to a suitable temperature, e.g. 100°C, using a suitable alcohol and a base, e.g. potassium tert-butoxide, in a solvent, e.g. NMP. Alternatively, Z is bromo and the desired group R is 12 In the case where Z is an alkyne, this conversion can be carried out by heating compound (iv) in a suitable solvent, such as THF, at a suitable temperature, such as 70°C, together with a suitable alkyne, copper iodide (I), a suitable base, such as Finnig base, and a suitable source of palladium, such as tetrakis(triphenylphosphine)palladium (O). Alternatively, Z is a trilate and the desired group R 12 In the case where is an alkyl group arbitrarily substituted, this step can be achieved by treating compound (iv) with a suitable alkyl boronic acid or ester, a catalyst such as a PdCl2(dppf)-DCM complex, and a base such as cesium carbonate in a solvent such as dioxane.

[0327] Step 5: In Step 5 of Reaction Scheme 1, compound (v) is converted to amine (vi) by incorporating a fluorosubstituent through treatment with a suitable electrophilic halogenating agent, such as selectfluorine, while stirring in a suitable solvent, such as tetrahydrofuran and acetonitrile, at room temperature. Additionally, electrophilic halogenating agents, such as NCS, NBS, and NIS, are applied to compound (vi) using a suitable solvent, such as THF, 1,4-dioxane, or acetonitrile, at room temperature or elevated temperature. 7a Cl, Br, or I substituents can be incorporated.

[0328] Steps 6 to 8 of Reaction Scheme 1 are substituent R of intermediate (v). 8 , R 9 , R 10 , R 11 , and R 12 The desired substituent R in the compound of chemical formula (I). 1 , R 2 , R 3 , R 4 It consists of a series of optional functional group manipulations to convert to , and W. A person skilled in the art will recognize that some or all of these steps may not be necessary depending on the groups present in compounds (v) and (ix). A person skilled in the art will also recognize that, in some descriptions, these steps may be performed in an alternative order.

[0329] Step 6: Step 6 of Reaction Scheme 1 is the gas R in compound (vi). 9 R present in molecule (vii) 2 It is an arbitrary step or series of steps for converting to. For example, R 9 A is Bromo and the intended R 2If A is an aromatic or heteroaromatic group, this conversion may be carried out by reacting compound (vi) with an optionally protected aromatic or heteroaromatic boronic acid or boronic acid ester in a solvent mixture such as dioxane and water, a catalyst such as a PdCl2(dppf)-DCM complex, and a base such as tripotassium phosphate. If the introduced group contains a protecting group, an additional optional step may be performed to remove the protecting group under appropriate conditions if desired. For example, if the introduced group is a pyrazole having a tetrahydropyran protecting group, the tetrahydropyran may be removed by reaction with an acid such as trifluoroacetic acid in a solvent such as dichloromethane. Alternatively, R 9 A is Bromo and the intended R 2 In the case where is an aromatic or heteroaromatic group, this conversion can be carried out by first reacting compound (vi) with a compound such as bis(pinacoleto)diborone, a reagent such as potassium acetate, and a catalyst such as a PdCl2(dppf)-DCM complex in a solvent such as dioxane, and then reacting the resulting boron acid ester with a suitable aryl or heteroaryl halide, a base such as sodium carbonate, and a catalyst such as tetrakis(triphenylphosphine)palladium (0) in a suitable solvent mixture such as dioxane and water. Alternatively, R 9 A is Bromo and the intended R 2 In the case where it is a heterocycle connected through nitrogen atoms, this step can be carried out by reacting compound (vi) with a suitable heterocycle in the presence of a copper source such as copper iodide (I), a base such as sodium carbonate, and a ligand such as N,N'-dimethylethane-1,2-diamine in a suitable solvent such as DMSO.

[0330] Step 7: Step 7 of Reaction Scheme 1 is the group R of intermediate (vii). 12It is an arbitrary step or a series of steps to convert into the group W present in molecule (viii). For example, group R 12 Where A contains a Boc-protected amine and the desired group W contains an amide, this conversion can be achieved by first removing the Boc group using a suitable combination of acid and solvent, such as hydrochloric acid and dioxane, and then forming the desired amide by reaction with a suitable carboxylic acid, a coupling agent, such as T3P, and a base, such as triethylamine, in a solvent, such as DMF. Alternatively, group R 12 When the group contains an unsaturated group, such as an alkyne, and the target group W is completely saturated, this conversion can be carried out by reaction with hydrogen and a suitable catalyst, such as carbon-phase palladium.

[0331] Step 8: Step 8 of Reaction Scheme 1 is the gas R in compound (viii). 8 , R 10 , and R 11 R present in the compound of chemical formula (I) 1 , R 3 , and R 4 It is an arbitrary step or series of steps to convert it into.

[0332] A person skilled in the art will recognize that many of these steps can be performed in an alternative order depending on the desired group in the compound of formula (I). For example, for some molecules, the group R described in step 6 9 of R 2 The conversion to is the R of Z described in Step 4. 12 It can be performed before the conversion to.

[0333] An alternative synthesis for the preparation of compound (vi) in Reaction Scheme 1 is presented in Reaction Scheme 2. In this case, R 7aStarting with a suitably functionalized quinolinol (x) that has been functionalized by [the method], follow steps 1-4 outlined in Reaction Scheme 1. Compound (vi) can be processed into the compound of formula (I) according to steps 6-8 outlined in Reaction Scheme 1.

[0334] Reaction Equation 2

[0335]

[0336] Chemical formula (I, R 7a The synthesis of compounds of (II), and (III), where (a) is an alkyl or cycloalkyl, can be carried out using the method summarized in Reaction Scheme 3.

[0337] Reaction Equation 3

[0338]

[0339] Step 1: The first step of Scheme 3 begins with suitably functionalized 2-aminobenzoate (xiii). If desired, group R 8 , R 9 , R 10 , and R 11 R present in the final product 1 , R 2 , R 3 , and R 4 It may be. Alternatively, one or more of these groups may be groups that can be modified in later stages of synthesis, such as bromo. Additionally, R 13 Eun Gi R 7 or R 7a It may be (alkyl or cycloalkyl). 2-aminobenzoate may be commercially available or synthesized by methods known to those skilled in the art. In step 1, the ester group of compound (xiii) may be converted to oxobutanitrile (xiv) under conditions such as substitution with a lithiate, such as acetonitrile, produced by the addition of a base, such as n-BuLi, in a solvent, such as THF.

[0340] Step 2: In Step 2 of Reaction Scheme 3, compound (xiv) can be converted to quinolinol (xv) through base-catalyzed cyclization by exposing (xiv) to a base, such as sodium ethoxide, in a solvent, such as ethanol, at a temperature as high as 100°C.

[0341] Alternatively, steps 1 and 2 can be performed in step 1 by allowing step 1 to continue at room temperature for an extended period.

[0342] Steps 3 to 7 of Reaction Scheme 3 are substituent R of the intermediate (xv). 8 , R 9 , R 10 , R 11 , R 13 , and OH as the desired substituent R in the compound of formula (II). 1a , R 2 , R 3 , R 4 , R 7 , and the desired R in W or the compound of formula (III). 1 , R 2 , R 3 , R 4a , R 7 It consists of some optional series of functional group manipulations to convert to , and W. A person skilled in the art will recognize that some or all of these steps may not be necessary depending on the groups present in compound (xv), or compounds of formulas (II) and (III). A person skilled in the art will also recognize that, in some cases, these steps may be performed in an alternative order.

[0343] Step 3: Step 3 of Reaction Scheme 3 is the gas R in compound (xv). 9 R present in molecule (xvi) 2 It is an arbitrary step or series of steps for converting to. For example, R 9 A is Bromo and the intended R 2If A is an aromatic or heteroaromatic group, this conversion may be carried out by reacting compound (xv) in a solvent mixture, such as dioxane and water, with an optionally protected aromatic or heteroaromatic boronic acid or boronic acid ester, a catalyst, such as a PdCl2(dppf)-DCM complex, and a base, such as tripotassium phosphate. If the introduced group contains a protecting group, an additional optional step may be performed to remove the protecting group under appropriate conditions if desired. For example, if the introduced group is a pyrazole having a tetrahydropyran protecting group, the tetrahydropyran may be removed by reaction with an acid, such as trifluoroacetic acid, in a solvent, such as dichloromethane. Alternatively, R 9 A is Bromo and the intended R 2 In the case where is an aromatic or heteroaromatic group, this conversion can be carried out by first reacting compound (xv) with a compound such as bis(pinacoleto)diborone, a reagent such as potassium acetate, and a catalyst such as a PdCl2(dppf)-DCM complex in a solvent such as dioxane, and then reacting the resulting boron acid ester with a suitable aryl or heteroaryl halide, a base such as sodium carbonate, and a catalyst such as tetrakis(triphenylphosphine)palladium (0) in a suitable solvent mixture such as dioxane and water. Alternatively, R 9 A is Bromo and the intended R 2 In the case where it is a heterocycle connected through nitrogen atoms, this step can be carried out by reacting the compound (xv) with a suitable heterocycle in the presence of a copper source such as copper iodide (I), a base such as sodium carbonate, and a ligand such as N,N'-dimethylethane-1,2-diamine in a suitable solvent such as DMSO.

[0344] Step 4: In Step 4 of Reaction Scheme 3, the alcohol group of compound (xvi) can be converted to a halogen group or a sulfonate ester, such as chloro, bromo, or triflate. If the desired group Z is chloro, this conversion can be carried out by treating compound (xvi) with a reagent, such as phosphoryl chloride, in a solvent, such as toluene. Alternatively, if the desired group Z is bromo, this conversion can be carried out by treating compound (xvi) with a reagent, such as tribromide, in a solvent, such as DMF. Alternatively, if the desired group Z is triflate, this conversion can be carried out by treating compound (xvi) with a reagent, such as trifluoromethanesulfonyl chloride, a reagent, such as 4-dimethylaminopyridine, and a base, such as Finnig base, in a solvent, such as dichloromethane.

[0345] Step 5: In Step 5 of Reaction Scheme 3, the halogen Z of compound (xvii) is replaced by the group R of compound (xviii). 12 Converts to. Gi R 12 may be the desired group W in the final compound; alternatively, it may be a group that can be converted to group W in later stages of synthesis. A person skilled in the art would know that the means for carrying out such conversion is group R 12 You will be aware that it will vary depending on the properties of Z. For example, if Z is chloro and the target group R 12 If Z is an amine, this conversion can be carried out by heating the compound (xvii) in a solvent, such as DMSO or NMP, with a suitable amine and a base, such as Finnig base, at a suitable temperature, such as 120°C. Alternatively, Z is chloro and the desired group R 12If α is an ether, this conversion can be carried out by heating compound (xvii) in a solvent such as NMP with a suitable alcohol and a base such as potassium tert-butoxide to a suitable temperature such as 100°C. Alternatively, if Z is bromo and the desired group R is 12 In the case where is an alkyne, this conversion can be carried out by heating the compound (xvii) in a suitable solvent, such as THF, with a suitable alkyne, copper iodide (I), a suitable base, such as Finnig base, and a suitable source of palladium, such as tetrakis(triphenylphosphine)palladium (O), to a suitable temperature, e.g., 70°C. Alternatively, Z is a trilate and the desired group R 12 In the case where is an alkyl group arbitrarily substituted, this step can be achieved by treating compound (xvii) with a suitable alkyl boronic acid or ester, a catalyst such as a PdCl2(dppf)-DCM complex, and a base such as cesium carbonate in a solvent such as dioxane.

[0346] Step 6: Step 6 of Equation 3 is the group R of the intermediate (xviii). 12 It is an arbitrary step or a series of steps to convert into the group W present in molecule (xix). For example, group R 12 Where A contains a Boc-protected amine and the desired group W contains an amide, this conversion can be achieved by first removing the Boc group using a suitable combination of acid and solvent, e.g., hydrochloric acid and dioxane, and then forming the desired amide by reaction with a suitable carboxylic acid, a coupling agent, e.g., T3P, and a base, e.g., triethylamine, in a solvent, e.g., DMF. Alternatively, group R 12 When the group contains an unsaturated group, such as an alkyne, and the target group W is completely saturated, this conversion can be carried out by reaction with hydrogen and a suitable catalyst, such as carbon-phase palladium.

[0347] Step 7: Step 7 of Reaction Scheme 3 is the gas R in compound (xix). 8 , R 10 , R 11 , and R 13 R present in the compound of chemical formula (II) 1a , R 3 , and R 4 , and R 7 or group R present in the compound of formula (III) 1 , R 3 , and R 4a , and R 7 It is an arbitrary step or series of steps to convert it into.

[0348] A person skilled in the art will recognize that many of these steps may be performed in an alternative order depending on the desired group in the compounds of formulas (II) and (III). For example, for some molecules, the group R described in step 3 9 of R 2 The conversion to is the R of Z described in Step 5. 12 It can be performed after conversion to.

[0349] An alternative approach to the synthesis of compounds of chemical formulas (II) and (III) can be carried out using the method summarized in reaction scheme 4.

[0350] Reaction Equation 4

[0351]

[0352] Step 1: Step 1 of Reaction Scheme 4 begins with suitablely functionalized aniline (xx). If desired, group R 8 , R 9 , R 10 , and R 11 R, present in the compound of chemical formula (II). 1a , R 2 , R 3 , and R 4 or group R present in the compound of formula (III) 1 , R2 , R 3 , and R 4a It may be. Alternatively, one or more of these groups may be groups that can be modified in later stages of synthesis, such as bromo. Aniline may be commercially available or synthesized by methods known to those skilled in the art. In step 1, the aniline of compound (xx) may be converted into compound (xxi) by treating it with commercially available 5-(methoxymethylene)-2,2-dimethyl-1,3-dioxane-4,6-dione in a solvent, such as diphenyl ether, when heated at a temperature, such as 120°C.

[0353] Step 2: In Step 2 of Reaction Scheme 4, compound (xxiv) can be converted to quinolinol (i) when heated in a solvent, such as diphenyl ether, at a temperature of 240°C.

[0354] Step 3: In Step 3 of Reaction Scheme 4, the alcohol group of compound (i) can be converted to a halogen group or a sulfonate ester, such as chloro, bromo, or triflate. If the desired group Z is chloro, this conversion can be carried out by treating compound (ii) with a reagent, such as phosphoryl chloride, in a solvent, such as toluene. Alternatively, if the desired group Z is bromo, this conversion can be carried out by treating compound (i) with a reagent, such as tribromide, in a solvent, such as DMF. Alternatively, if the desired group Z is triflate, this conversion can be carried out by treating compound (ii) with a reagent, such as trifluoromethanesulfonyl chloride, a reagent, such as 4-dimethylaminopyridine, and a base, such as Finnig base, in a solvent, such as dichloromethane.

[0355] Step 4: In Step 4 of Reaction Scheme 4, compound (ii) is treated with a suitable oxidizing agent, such as meta-chloroperoxybenzoic acid, in a solvent such as DCM to convert it into N-oxide (iii).

[0356] Step 5: In Step 5 of Reaction Scheme 4, compound (iii) is treated in a suitable solvent, such as DCM, with a suitable activating reagent, such as tosyl chloride, and an ammonia source, such as ammonium chloride and triethylamine, to convert it into amine (iv).

[0357] Step 6: In Step 6 of Reaction Scheme 4, the halogen Z of compound (iv) is replaced by the group R of compound (v). 12 Converts to. Gi R 12 may be the desired group W in the final compound; alternatively, it may be a group that can be converted to group W in later stages of synthesis. A person skilled in the art would know that the means for carrying out such conversion is group R 12 You will be aware that it will vary depending on the properties of Z. For example, if Z is chloro and the target group R 12 In the case where α is an amine, this conversion can be carried out by heating compound (iv) to a suitable temperature, e.g. 120°C, in a solvent, e.g. DMSO or NMP, with a suitable amine and a base, e.g. Finnig base. Alternatively, Z is chloro and the desired group R is 12 In the case where α is an ether, this conversion can be carried out by heating compound (iv) in a solvent such as NMP with a suitable alcohol and a base such as potassium tert-butoxide to a suitable temperature, e.g., 100°C. Alternatively, Z is bromo and the desired group R is 12 In the case where is an alkyne, this conversion can be carried out by heating compound (iv) in a suitable solvent, such as THF, with a suitable alkyne, copper iodide (I), a suitable base, such as Finnig base, and a suitable source of palladium, such as tetrakis(triphenylphosphine)palladium (O), to a suitable temperature, such as 70°C. Alternatively, Z is a trilate and the desired group R 12In the case where is an alkyl group arbitrarily substituted, this step can be achieved by treating compound (iv) with a suitable alkyl boronic acid or ester, a catalyst such as a PdCl2(dppf)-DCM complex, and a base such as cesium carbonate in a solvent such as dioxane.

[0358] Step 7: Step 7 of Equation 4 is the gas R of intermediate (v). 9 R present in molecule (xxii) 2 It is an arbitrary step or series of steps for converting to. For example, R 9 A is Bromo and the intended R 2 If A is an aromatic or heteroaromatic group, this conversion may be carried out by reacting compound (v) with an optionally protected aromatic or heteroaromatic boronic acid or boronic acid ester in a solvent mixture such as dioxane and water, a catalyst such as a PdCl2(dppf)-DCM complex, and a base such as tripotassium phosphate. If the introduced group contains a protecting group, an additional optional step may be performed to remove the protecting group under appropriate conditions if desired. For example, if the introduced group is a pyrazole having a tetrahydropyran protecting group, the tetrahydropyran may be removed by reaction with an acid such as trifluoroacetic acid in a solvent such as dichloromethane. Alternatively, R 9 A is Bromo and the intended R 2 In the case where is an aromatic or heteroaromatic group, this conversion can be carried out by first reacting compound (v) with a compound such as bis(pinacoleto)diborone, a reagent such as potassium acetate, and a catalyst such as a PdCl2(dppf)-DCM complex in a solvent such as dioxane, and then reacting the resulting boron acid ester with a suitable aryl or heteroaryl halide, a base such as sodium carbonate, and a catalyst such as tetrakis(triphenylphosphine)palladium (0) in a suitable solvent mixture such as dioxane and water. Alternatively, R 9A is Bromo and the intended R 2 In the case where it is a heterocycle connected through nitrogen atoms, this step can be carried out by reacting compound (v) with a suitable heterocycle in the presence of a copper source such as copper iodide (I), a base such as sodium carbonate, and a ligand such as N,N'-dimethylethane-1,2-diamine in a suitable solvent such as DMSO.

[0359] Step 8: Step 8 of Reaction Scheme 4 is the group R in compound (xxii). 12 It is an optional step or a series of steps to convert into the group W present in the compound of formula (II) or (III). For example, group R 12 Where A contains a Boc-protected amine and the desired group W contains an amide, this conversion can be achieved by first removing the Boc group using a suitable combination of acid and solvent, such as hydrochloric acid and dioxane, and then forming the desired amide by reaction with a suitable carboxylic acid, a coupling agent, such as T3P, and a base, such as triethylamine, in a solvent, such as DMF. Alternatively, group R 12 If G contains an unsaturated group, such as an alkyne, and the desired group W is completely saturated, this conversion can be carried out by reaction with hydrogen and a suitable catalyst, such as carbon-phase palladium. Additionally, the group R in compound (xxii) 8 , R 10 , and R 11 R present in the compound of chemical formula (II) 1a , R 3 and R 4 or group R present in the compound of formula (III) 1 , R 3 , and R 4a A series of arbitrary steps to convert it can be performed.

[0360] A person skilled in the art will recognize that many of these steps may be performed in an alternative order depending on the desired group in the compounds of formulas (II) and (III). For example, for some molecules, the group R described in step 7 9 of R 2 The conversion to is the R of Z described in Step 6. 12 It can be performed before the conversion to.

[0361] Chemical formula (I, R 7a The approach to the synthesis of compounds (where alkyl or cycloalkyl) can be carried out using the method summarized in Reaction Scheme 5.

[0362] Reaction Equation 5

[0363]

[0364] Step 1: Step 1 of Reaction Scheme 6 is the gas R in compound (iv). 9 The group R present in molecule (xxiii) 2 It is an arbitrary step or series of steps for converting to. For example, R 9 A is Bromo and the intended R 2 If g is an aromatic or heteroaromatic group, this conversion may be carried out by reacting compound (iv) in a solvent mixture, such as dioxane and water, with an optionally protected aromatic or heteroaromatic boronic acid or boronic acid ester, a catalyst, such as a PdCl2(dppf)-DCM complex, and a base, such as tripotassium phosphate. If the introduced group contains a protecting group, an additional optional step may be performed to remove the protecting group under appropriate conditions if desired. For example, if the introduced group is a pyrazole having a tetrahydropyran protecting group, the tetrahydropyran may be removed by reaction with an acid, such as trifluoroacetic acid, in a solvent, such as dichloromethane. Alternatively, R 9 A is Bromo and the intended R 2In the case where is an aromatic or heteroaromatic group, this conversion can be carried out by first reacting compound (iv) with a compound such as bis(pinacoleto)diborone, a reagent such as potassium acetate, and a catalyst such as a PdCl2(dppf)-DCM complex in a solvent such as dioxane, and then reacting the resulting boron acid ester with a suitable aryl or heteroaryl halide, a base such as sodium carbonate, and a catalyst such as tetrakis(triphenylphosphine)palladium (0) in a suitable solvent mixture such as dioxane and water. Alternatively, R 9 A is Bromo and the intended R 2 In the case where it is a heterocycle connected through nitrogen atoms, this step can be carried out by reacting compound (iv) with a suitable heterocycle in the presence of a copper source such as copper iodide (I), a base such as sodium carbonate, and a ligand such as N,N'-dimethylethane-1,2-diamine in a suitable solvent such as DMSO.

[0365] Step 2: In Step 2 of Reaction Scheme 5, compound (xxiii) is converted to amine (xxiv) by treating it with a suitable alkyl or cycloalkyl trifluoroborate salt in the presence of manganese(III) acetate dihydrate in a suitable solvent mixture, such as acetic acid-water, heated to a temperature, such as 50°C.

[0366] Step 3: In Step 3 of Reaction Scheme 5, the halogen Z of compound (xxiv) is replaced by the group R of compound (xxv). 12 Converts to. Gi R 12 may be the desired group W in the final compound; alternatively, it may be a group that can be converted to group W in later stages of synthesis. A person skilled in the art would know that the means for carrying out such conversion is group R 12 You will be aware that it will vary depending on the properties of Z. For example, if Z is chloro and the target group R 12If Z is an amine, this conversion can be carried out by heating the compound (xxiv) in a solvent, such as DMSO or NMP, with a suitable amine and a base, such as Finnig base, to a suitable temperature, e.g., 120°C. Alternatively, Z is chloro and the desired group R 12 If α is an ether, this conversion can be carried out by heating compound (xxiv) in a solvent such as NMP with a suitable alcohol and a base such as potassium tert-butoxide to a suitable temperature, e.g., 100°C. Alternatively, if Z is bromo and the desired group R is 12 In the case where is an alkyne, this conversion can be carried out by heating the compound (xxiv) in a suitable solvent, such as THF, with a suitable alkyne, copper iodide (I), a suitable base, such as Finnig base, and a suitable palladium source, such as tetrakis(triphenylphosphine)palladium (O), to a suitable temperature, e.g. 70°C. Alternatively, Z is a trilate and the desired group R 12 In the case where is an alkyl group arbitrarily substituted, this step can be achieved by treating compound (xxiv) with a suitable alkyl boronic acid or ester, a catalyst such as a PdCl2(dppf)-DCM complex, and a base such as cesium carbonate in a solvent such as dioxane.

[0367] Step 4: Step 4 of Reaction Scheme 5 is the gas R of the intermediate (xxv). 12 It is an arbitrary step or series of steps to convert into the gas W present in molecule (xxvi). For example, gas R 12 Where A contains a Boc-protected amine and the desired group W contains an amide, this conversion can be achieved by first removing the Boc group using a suitable combination of acid and solvent, such as hydrochloric acid and dioxane, and then forming the desired amide by reaction with a suitable carboxylic acid, a coupling agent, such as T3P, and a base, such as triethylamine, in a solvent, such as DMF. Alternatively, group R12 When the group contains an unsaturated group, such as an alkyne, and the target group W is completely saturated, this conversion can be carried out by reaction with hydrogen and a suitable catalyst, such as carbon-phase palladium.

[0368] Step 5: Step 5 of Reaction Scheme 5 is the gas R in compound (xxvi). 8 , R 10 , and R 11 R present in the compound of chemical formula (I) 1 , R 3 , and R 4 It is an arbitrary step or series of steps to convert it into.

[0369] A person skilled in the art will recognize that many of these steps may be performed in an alternative order depending on the desired group in the compound of formula (I). For example, for some molecules, the group R described in step 1 9 of R 2 The conversion to is the R of Z described in Step 4. 12 It can be performed after conversion to.

[0370] Evaluation of biological activity

[0371] Measurement of IL-1β production in PMA-differentiated THP-1 cells

[0372] THP-1 cells were purchased from the American Type Culture Collection and subcultured according to the supplier's instructions. Prior to the experiment, cells were cultured in RPMI 1640 containing 10% heat-inactivated FBS, penicillin (100 units / ml), and streptomycin (100 μg / ml) and maintained in the logarithmic phase prior to experimental setup. Prior to the experiment, THP-1 was treated with PMA (Phorbol 12-myristate 13-acetate) (10 μg / ml) for 24 hours. On the day of the experiment, the medium was removed, adhered cells were treated with trypsin for 2 minutes, followed by cell collection, washing with PBS (phosphate-buffered saline), spin-down, and 1 x 10⁶ cells in RPMI containing 2% heat-inactivated FBS. 6 The cells were resuspended at a concentration of 1 cell / ml and 100 μl was plated into a 96-well plate. The compound was dissolved in dimethyl sulfoxide (DMSO) and added to the culture medium to achieve the desired concentration (e.g., 100, 30, 10, 3, 1, 0.3, or 0.1 μM). Cells were incubated with the compound for 4 hours. Cell-free supernatants were collected, and IL-1β production was evaluated by ELISA. A vehicle-only control was tested concurrently with each experiment. The final DMSO concentration was 1%. The compound demonstrated a dose-related increase in IL-1β production in PMA-differentiated THP-1 cells.

[0373] Measurement of IL-1β production in PMA-differentiated THP-1 cells (Alternative procedure)

[0374] THP-1 cells were purchased from the American Type Culture Collection and subcultured according to the supplier's instructions. Prior to the experiment, cells were cultured in RPMI 1640 containing 10% heat-inactivated FBS, penicillin (100 units / ml), streptomycin (100 μg / ml), HEPES (10 mM), and sodium pyruvate (1 mM), and maintained in the logarithmic phase prior to experimental setup. Prior to the experiment, THP-1 cells were treated overnight with PMA (Porbol 12-Myristate 13-Acetate) (20 μg / ml). On the day of the experiment, the medium was removed, adhered cells were treated with trypsin for 2 minutes, followed by cell collection, washing with PBS (phosphate-buffered saline), pelleting by centrifugation, and resuspending in RPMI containing 2% heat-inactivated FBS in 384-well plates at a concentration of 50,000 cells / well. Cell-free supernatants were collected, and IL-1β production was evaluated by ELISA. The compound was dissolved in dimethyl sulfoxide (DMSO) and added to culture medium to achieve target concentrations (e.g., 100, 30, 10, 3, 1, 0.3, or 0.1 μM). Cells were incubated with the compound for 2 hours. A vehicle-only control was tested concurrently with each experiment. The final DMSO concentration was 1%. The compound demonstrated a dose-related increase in IL-1β production in PMA-differentiated THP-1 cells.

[0375] Measurement of IL-1β Production - hTRF Protocol (Second Alternative Procedure)

[0376] Serial dilutions of the compound in DMSO were added at a rate of 100 nl / well to a low-volume 384-well plate using an ECHO 550 acoustic splitter (Labcyte) to achieve a final starting concentration of 10 μM in the assay.

[0377] 1 x 10⁶ in RPMI (Gibco, 11875) medium containing 10% FBS in a T175 flask6 THP-1 cells at a density of 10 cells / ml were treated with a final concentration of 50 ng / ml of phorbol 12-myristate 13-acetate (PMA) (Sigma, P1585) overnight at 37°C in 5% CO2 for differentiation. The next day, cells were harvested after rinsing the wells with dPBS using 0.5% trypsin. 1x10 6 A cell solution of 1 cell / ml was prepared and mixed with 50 μl / well in RPMI medium containing 2% FBS to obtain 50,000 cells. Using a multichannel pipette, the cells were plated onto the compound dilution in a Greiner 384-well, black clear-bottomed tissue culture plate (781090). The plate was incubated in a 37°C incubator at 5% CO2 for 2 hours.

[0378] After 2 hours of incubation, the cell plates were centrifuged at 1200 rpm for 5 minutes. Using Felix (CyBio), 8 μl of the supernatant was transferred to a 384-well, low-volume, white proxy plate (Perkin Elmer, 6008230). The supernatant was analyzed using the Human IL1-beta hTRF Kit (CISBIO, 62HIL1BPEG). An IL1-beta standard curve was constructed according to the kit instructions, and the antibody from the kit was subsequently diluted to 1:40 instead of the 1:20 indicated on the kit. After aggregation, the antibody was added to the plates at a rate of 5 μl / well. The plates were sealed and incubated overnight at 4°C. Subsequently, the plates were read on a Perkin Elmer Envision at 665 / 615 nm using an hTRF laser. The compound showed a capacity-related increase in IL-1β production.

[0379] Measurement of IL-1β Production - Human Whole Blood Test

[0380] Serial dilutions of the compound in DMSO were added to a low-volume 384-well plate at 100 nl / well using an ECHO 550 acoustic splitter (LabSight) to achieve a final starting concentration of 10 uM in the assay.

[0381] Human venous whole blood obtained from healthy donors was pretreated with 1 ng / ml LPS (Invivogen, Cat# tlrl-eblps) for 4 hours at 37°C in a humidified 95% air / 5% CO2 incubator. Primed blood was added to compound plates and incubated for an additional 4 hours at 37°C. IL-1β in the supernatant was measured using the AlphLISA kit (Cat# AL220) according to the manufacturer's instructions. The compound showed a dose-related increase in IL-1β production. EC50 was determined using primed but untreated blood as a baseline.

[0382] Measurement of IL-1β Production - Mouse hTRF Protocol

[0383] Immortalized mouse macrophages derived from C57BL / 6 mice were obtained from Ericke Latz at the University of Massachusetts / Worcester, Massachusetts. Cells were harvested using 0.05% trypsin and washed with PBS. Cells were plated at 30,000 cells per well in 25 µl of DMEM (Gibco, 11965) supplemented with 2% FBS and incubated at 37°C for 10 minutes in 5% CO2. LPS-EB (Invivogen, tlr-eblps) was added at a final concentration of 200 ng / ml at 5 µl / well, and cells were incubated at 37°C for 2 hours in 5% CO2.

[0384] Serial dilutions of the compound in DMSO were added to cells in a low-volume 384-well plate at 60 nl / well using an ECHO 550 acoustic splitter (Labsite) to achieve a final starting concentration of 50 uM in the assay, and incubated with the compound for an additional 2 hours at 37°C in 5% CO2.

[0385] After 2 hours of incubation, the cell plates were centrifuged at 1200 rpm for 5 minutes. Using Felix (Cybio), 8 μl of the supernatant was transferred to a 384-well, low-volume, white proxy plate (PerkinElmer, 6008230). The supernatant (CysBio, 62MIL1BPEH) was analyzed using the Human IL1beta hTRF Kit. An IL1beta standard curve was constructed according to the kit instructions (the antibody from the kit was diluted to 1:40 instead of the 1:20 indicated on the kit). After aggregation, the antibody was added throughout the plate at a rate of 5 μl / well. The plates were sealed and incubated overnight at 4°C. The plates were read on a PerkinElmer Envision at 665 / 615 nm using an hTRF laser. Subsequently, the data were converted to IL1beta in pg / ml. The compound showed a capacity-related increase in IL-1β production.

[0386] In vitro human TLR7 and TLR8 binding reporter assay

[0387] Logarithmically growing human HEK-Blue cells co-expressing the TLR7 or TLR8 gene and the NF-kB / AP1-inducible SEAP (secreted embryonic alkaline phosphatase; Invivogen, San Diego, California) reporter gene were added to individual wells of a 384-well plate (15,000 cells per 20 μL well) and maintained at 37°C, 5% CO2 for 24 hours. The test compound or DMSO was dispensed into individual wells the following day using an acoustic liquid handling technique (100 nL per well), and the cells were subsequently incubated at 37°C, 5% CO2 for 18 hours. Newly prepared Quanti-Blue reagent (prepared according to manufacturer's instructions; Invivogen, San Diego, California) was added to the HEK-Blue TLR Nf-kB-SEAP cell reaction, and after 30 minutes, cell SEAP production was measured using an Envision plate reader. All EC 50 The value (half-maximum effective concentration) was determined using dedicated data analysis software. Normalized EC 50 Value = Absolute value determined by setting 100% Ymax using the reference standard RLU (Relative Light Unit) value from cells treated with a 50 μM reference standard.

[0388] Examples

[0389] To further illustrate the foregoing, the following non-limiting and exemplary synthetic reaction formulas are included. Variations of these examples within the scope of the claims are within the understanding of those skilled in the art and are deemed to fall within the scope of the invention as described and claimed herein. Readers will recognize that a person skilled in the art who has received this disclosure can manufacture and use the invention without complete embodiments.

[0390] The biological data of the compounds were verified using one or more of the above procedures. Unless otherwise indicated, the TLR7 agonist EC of the following compounds50 and TLR8 agonist EC 50 It was measured at a value of >100 μM.

[0391] HPLC / MS and HPLC methods for purification / analysis used for characterizing or purifying examples

[0392] Analytical HPLC / MS was performed using the following method:

[0393] Method A: Column: Aquiti UPLC BEH C18, 1.7 μm particles; Mobile phase A: 99.95:0.05 Water:TFA; Mobile phase B: 99.95:0.05 Acetonitrile:TFA; Temperature: 50°C; Gradient: 2%B to 98%B over 1.00 min, followed by holding at 98%B for 0.50 min; Flow rate: 0.8 mL / min; Detection: MS and UV (254 nm).

[0394] Method B: Column: Waters Exbridge C18, 2.1 mm x 50 mm, 1.7 μm particles; Mobile Phase A: 5:95 acetonitrile:water, containing 10 mM ammonium acetate; Mobile Phase B: 95:5 acetonitrile:water, containing 10 mM ammonium acetate; Temperature: 50°C; Gradient: 0%B to 100%B over 3 minutes, followed by holding at 100%B for 0.50 minutes; Flow Rate: 1 mL / min; Detection: MS and UV 220 nm;

[0395] Method C: Column: Waters Exbridge C18, 2.1 mm x 50 mm, 1.7 μm particles; Mobile phase A: 5:95 acetonitrile:water, containing 0.1% trifluoroacetic acid; Mobile phase B: 95:5 acetonitrile:water, containing 0.1% trifluoroacetic acid; Temperature: 50°C; Gradient: 0% B to 100% B over 3 minutes, followed by holding at 100% B for 0.50 minutes; Flow rate: 1 mL / min; Detection: MS and UV (220 nm).

[0396] Nuclear Magnetic Resonance (NMR) Spectroscopy

[0397] Chemical shifts are recorded as parts per million (ppm) downfields from the internal tetramethylsilane (TMS) or from the position of TMS inferred by the deuterated NMR solvent. Apparent multiplicity is recorded as follows: single-s, double-d, triple-t, quadruple-q, or multiply-m. Peaks indicating broadening are additionally denoted by br. Integrations are approximate. It should be noted that integration intensity, peak shape, chemical shift, and coupling constant may vary depending on the solvent, concentration, temperature, pH, and other factors. Additionally, peaks that overlap or are exchanged with water or solvent peaks in the NMR spectrum may not provide reliable integration intensity. In some cases, NMR spectra may be obtained using water peak suppression, which may result in the overlapping peaks being invisible or altering their shape and / or integration.

[0398] Example 1. 3-((2-amino-3-fluoro-7-(1H-pyrazole-3-yl)quinoline-4-yl)amino)propan-1-ol

[0399]

[0400] 1A. 7-Bromo-4-chloroquinoline

[0401]

[0402] POCl3 (2.080 mL, 22.32 mmol) was added to a suspension of 7-bromoquinoline-4-ol (2.5 g, 11.16 mmol) in toluene (20 mL). The reaction mixture was heated to 100°C. After 1.5 hours, the reaction mixture was cooled, followed by the addition of ice. The reaction mixture was vigorously stirred for approximately 30 minutes, followed by the addition of water. The reaction mixture was extracted twice with DCM. The organic layer was washed with saturated aqueous NaHCO3 and brine, dried over sodium sulfate, and concentrated. A saturated aqueous NaHCO3 solution was slowly added to the aqueous layer while stirring. The precipitate was collected by vacuum filtration, washed with water, and dried. The solid was isolated from the organic layer, the filtered solid was combined, and dried under reduced pressure to obtain 7-bromo-4-chloroquinoline (2.46 g, 10.14 mmol, 91% yield).

[0403] 1 ¹H NMR (400 MHz, chloroform-d) δ 8.80 (d, J=4.7 Hz, 1H), 8.33 (d, J=1.9 Hz, 1H), 8.12 (d, J=9.0 Hz, 1H), 7.75 (dd, J=9.0, 2.0 Hz, 1H), and 7.52 (d, J=4.8 Hz, 1H).

[0404] 1B. 7-Bromo-4-chloroquinoline 1-oxide

[0405]

[0406] mCPBA (6.10 g, 24.74 mmol) was added to a solution of 7-bromo-4-chloroquinoline (2.0 g, 8.25 mmol) in DCM (55.0 mL). The reaction mixture was stirred overnight and then quenched with a saturated sodium thiosulfate solution. The reaction mixture was stirred for 0.5 hours and a saturated aqueous sodium bicarbonate solution was added. The reaction mixture was extracted twice with DCM, the organic layer was washed with brine, dried with sodium sulfate, and concentrated to obtain 7-bromo-4-chloroquinoline 1-oxide (2.16 g, 8.36 mmol, quantitative yield).

[0407] 1 ¹H NMR (400 MHz, chloroform-d) δ 8.99 (d, J=1.9 Hz, 1H), 8.43 (d, J=6.6 Hz, 1H), 8.10 (d, J=9.0 Hz, 1H), 7.86 (dd, J=9.0, 2.0 Hz, 1H), and 7.40 (d, J=6.6 Hz, 1H).

[0408] 1C. 7-Bromo-4-chloroquinoline-2-amine

[0409]

[0410] In one round-bottom flask, 7-bromo-4-chloroquinoline 1-oxide (9400 mg, 36.4 mmol) was suspended in DCM (150 mL). Ts-Cl (7626 mg, 40.0 mmol) was added. The mixture was stirred for 1 hour. In a second round-bottom flask, ammonium chloride (9725 mg, 182 mmol) (dried overnight in an oven at 110°C) was suspended in DCM (150 mL). Triethylamine (25.3 mL, 182 mmol) was added, and the mixture was stirred for 0.5 hours. The contents of the first flask were added to the second flask, the reaction mixture was stirred overnight, filtered, and concentrated. The residue was dissolved in 100 mL of hot DCM. The solution was cooled to room temperature, and the solid was collected by vacuum filtration. The filter cake was washed with 100 mL of DCM (-20°C). The filter cake was suspended in water (50 mL) and filtered. The DCM filtrate was evaporated, the residue was suspended in water (100 mL) and filtered. The filter cake was washed with 100 mL of DCM (-20°C) to obtain additional products. The combined solids were dried under reduced pressure to obtain 7-bromo-4-chloroquinoline-2-amine (6.52 g, 70% yield).

[0411] 1 H NMR (400 MHz, DMSO-d6) δ 7.79 (d, J=8.7 Hz, 1H), 7.65 (d, J=1.9 Hz, 1H), 7.39 (dd, J=8.8, 2.0 Hz, 1H), 6.98 (s, 1H), and 6.88 (s, 2H).

[0412] 1D. 3-((2-amino-7-bromoquinoline-4-yl)amino)propan-1-ol

[0413]

[0414] A mixture of 7-bromo-4-chloroquinoline-2-amine (0.200 g, 0.777 mmol), 3-aminopropan-1-ol (0.594 mL, 7.77 mmol), Finnig base (0.678 mL, 3.88 mmol), and DMSO (0.8 mL) in a screw-stop vial of 2 Drams was heated overnight in an oil bath at 120°C. The reaction mixture was diluted with dichloromethane, washed with 10% aqueous lithium chloride (2x), washed with brine, and dried over anhydrous sodium sulfate. After concentrating under reduced pressure, the product was subsequently purified by ISCO silica gel chromatography (12 g; 0-30% methanol in dichloromethane) to obtain 3-((2-amino-7-bromoquinoline-4-yl)amino)propan-1-ol (0.161 g, 0.538 mmol, 69% yield) as a white solid.

[0415] LC / MS [M+H] + = 296.2 and 298.2.

[0416] 1E. 3-((2-amino-7-bromo-3-fluoroquinoline-4-yl)amino)propan-1-ol

[0417]

[0418] A mixture of 3-((2-amino-7-bromoquinoline-4-yl)amino)propan-1-ol (0.108 g, 0.365 mmol) and selectfluoro (0.142 g, 0.401 mmol) in a mixture of tetrahydrofuran (0.5 mL) and acetonitrile (2.5 mL) was stirred at room temperature for 4 hours. The reaction mixture was concentrated under reduced pressure, the residue was diluted with dichloromethane, and washed with 1N aqueous sodium hydroxide (2x). The organic layer was collected, and the pH of the aqueous layer was adjusted to ~9 using aqueous sodium hydroxide. The mixture was extracted with ethyl acetate (3x). The organic layer was washed with brine and dried over anhydrous sodium sulfate. 3-((2-amino-7-bromo-3-fluoroquinoline-4-yl)amino)propan-1-ol (11 mg, 0.035 mmol, 10% yield) was obtained as a white solid by concentrating under reduced pressure.

[0419] LC / MS [M+H] + = 314.0 and 216.0.

[0420] Example 1

[0421] A mixture of 3-((2-amino-7-bromo-3-fluoroquinoline-4-yl)amino)propan-1-ol (0.011 g, 0.035 mmol), 3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazole (0.014 g, 0.070 mmol), and tripotassium phosphate (2 M in water) (0.053 mL, 0.105 mmol) was degassed (3x; vacuum / nitrogen) in dioxane (1.0 mL). 1,1'-bis(di-tert-butylphosphino)ferrocene palladium dichloride (2.86 mg, 3.50 μmol) was added to the mixture, and the mixture was degassed (3x; vacuum / nitrogen). The reaction mixture was immersed in an oil bath at 85°C and stirred overnight. The reaction mixture was diluted with ethyl acetate, washed with water, and washed with brine. The organic layer was collected, and the aqueous layer was sequentially extracted with ethyl acetate (3x). The combined organic layer was dried over anhydrous sodium sulfate, concentrated under reduced pressure, and the residue was purified by reverse-phase HPLC. The target fraction was concentrated and dried, the residue was dissolved in ethyl acetate, washed with a saturated aqueous solution of sodium bicarbonate, and then washed with brine. The organic layer was collected, and the aqueous layer was sequentially extracted with ethyl acetate (2x). The combined organic layer was concentrated to obtain 3-((2-amino-3-fluoro-7-(1H-pyrazole-3-yl)quinoline-4-yl)amino)propan-1-ol (3.2 mg, 10.51 μmol, 30% yield) as a white solid.

[0422] Example 2. (1S,3S)-3-((2-amino-3-fluoro-7-(1H-pyrazole-3-yl)quinoline-4-yl)amino)cyclopentan-1-ol

[0423]

[0424] 2A. (1S,3S)-3-((2-amino-7-bromoquinoline-4-yl)amino)cyclopentan-1-ol

[0425]

[0426] 2. In a screw-stop vial of Dram, a mixture of 7-bromo-4-chloroquinoline-2-amine (0.215 g, 0.835 mmol), (1S,3S)-3-aminocyclopentan-1-ol, HCl (0.300 g, 2.179 mmol), Finnig base (0.729 mL, 4.17 mmol), and DMSO (0.9 mL) was heated overnight in an oil bath at 120°C. The reaction mixture was diluted with dichloromethane, washed with 10% aqueous lithium chloride (2x), washed with brine, and dried over anhydrous sodium sulfate. After concentrating under reduced pressure, the (1S,3S)-3-((2-amino-7-bromoquinoline-4-yl)amino)cyclopentan-1-ol (0.208 g, 0.626 mmol, 75% yield) was subsequently purified by ISCO silica gel chromatography (12 g; 0-30% methanol in dichloromethane) to obtain (1S,3S)-3-((2-amino-7-bromoquinoline-4-yl)amino)cyclopentan-1-ol (0.208 g, 0.626 mmol, 75% yield) as a pale yellow solid.

[0427] LC / MS [M+H] + = 322.3 and 324.3.

[0428] 1 H NMR (400 MHz, DMSO-d6) δ 7.97 - 7.88 (m, 1H), 7.44 - 7.37 (m, 1H), 7.14 - 7.07 (m, 1H), 6.49 - 6.38 (m, 1H), 6.11 - 5.98 (m, 2H), 5.79 - 5.72 (m, 1H), 4.61 - 4.52 (m, 1H), 4.31 - 4.22 (m, 1H), 4.04 - 3.89 (m, 1H), 2.26 - 2.13 (m, 1H), 2.04 - 1.86 (m, 2H), 1.85 - 1.78 (m, 1H), and 1.63 - 1.47 (m, 2H).

[0429] 2B. (1S,3S)-3-((2-amino-7-bromo-3-fluoroquinoline-4-yl)amino)cyclopentan-1-ol

[0430]

[0431] A homogeneous mixture of (1S,3S)-3-((2-amino-7-bromoquinoline-4-yl)amino)cyclopentan-1-ol (0.104 g, 0.323 mmol) and selectfluoride (0.137 g, 0.387 mmol) in a mixture of tetrahydrofuran (1 mL) and acetonitrile (5 mL) was stirred overnight at room temperature. The solvent was removed under reduced pressure, the residue was sonicated in dichloromethane, and stirred overnight at room temperature. The dichloromethane layer was gently decanted, the residue was washed with dichloromethane, and the residue was gently decanted. The combined organic layer did not appear to contain the target product upon analysis, but the residue was a 1:1 mixture of the product and the starting material. The residue was purified by ISCO silica gel chromatography (12 g; 0-20% methanol in dichloromethane) to obtain (1S,3S)-3-((2-amino-7-bromo-3-fluoroquinoline-4-yl)amino)cyclopentan-1-ol (11 mg, 0.032 mmol, 10% yield) as a white solid.

[0432] LC / MS [M+H] + = 340.3 and 342.3.

[0433] 1 ¹H NMR (400 MHz, methanol-d4) δ 7.90 - 7.80 (m, 1H), 7.60 (s, 1H), 7.33 - 7.24 (m, 1H), 4.69 - 4.58 (m, 1H), 4.48 - 4.36 (m, 1H), 2.37 - 2.23 (m, 1H), 2.18 - 2.06 (m, 2H), 2.01 - 1.88 (m, 1H), and 1.78 - 1.58 (m, 2H).

[0434] Example 2

[0435] A mixture of (1S,3S)-3-((2-amino-7-bromo-3-fluoroquinoline-4-yl)amino)cyclopentan-1-ol (0.011 g, 0.032 mmol), 3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazole (0.013 g, 0.065 mmol), and tripotassium phosphate (2 M in water, 0.049 mL, 0.097 mmol) was degassed (3x; vacuum / nitrogen). 1,1'-bis(di-tert-butylphosphino)ferrocene palladium dichloride (2.64 mg, 3.23 μmol) was added to the mixture, and the mixture was degassed (3x; vacuum / nitrogen). The reaction mixture was immersed in an oil bath at 85°C and stirred overnight. The reaction mixture was diluted with ethyl acetate, washed with water, and washed with brine. The organic layer was collected, and the aqueous layer was sequentially extracted with ethyl acetate (3x). The combined organic layer was dried over anhydrous sodium sulfate, concentrated under reduced pressure, and the residue was purified by reverse-phase HPLC. The target fraction was concentrated and dried, the residue was dissolved in ethyl acetate, washed with a 1.5 M aqueous solution of dibasic potassium phosphate, and then washed with brine. The organic layer was collected, and the aqueous layer was sequentially extracted with ethyl acetate (2x). The combined organic layer was concentrated to obtain (1S,3S)-3-((2-amino-3-fluoro-7-(1H-pyrazole-3-yl)quinoline-4-yl)amino)cyclopentan-1-ol (6.4 mg, 0.019 mmol, 58.6% yield) as a grayish-white solid.

[0436] Example 3. 3-((2-amino-3-chloro-7-(1H-pyrazole-3-yl)quinoline-4-yl)amino)propan-1-ol

[0437]

[0438] 3A. 7-Bromo-3,4-Dichloroquinoline 1-Oxide

[0439]

[0440] Methyltrioxorenium (vii) (0.036 g, 0.144 mmol) was added to a solution of 7-bromo-3,4-dichloroquinoline (0.4 g, 1.444 mmol) in DCM (9.63 mL). The reaction mixture was placed in an ice bath, and hydrogen peroxide (35% in water) (0.379 ml, 4.33 mmol) was added. The mixture was stirred overnight. An additional 18 mg of methyltrioxorenium and hydrogen peroxide (35% in water) (0.379 ml, 4.33 mmol) were added. The reaction mixture was diluted with water, and a saturated sodium thiosulfate solution was added to generate vigorous bubbling. The reaction mixture was combined with the previous reaction mixture and extracted with DCM (3x). The organic layer was washed with a sodium bicarbonate solution, washed with brine, dried over sodium sulfate, and concentrated to obtain 7-bromo-3,4-dichloroquinoline 1-oxide (442 mg, 1.509 mmol, 87% yield).

[0441] LC / MS [M+H] + = 293.8.

[0442] 1 ¹H NMR (400 MHz, chloroform-d) δ 8.93 (d, J=2.0 Hz, 1H), 8.57 (s, 1H), 8.11 (d, J=8.9 Hz, 1H), and 7.88 (dd, J=8.9, 2.0 Hz, 1H).

[0443] 3B. 7-Bromo-3,4-dichloroquinoline-2-amine

[0444]

[0445] Ts-Cl (157 mg, 0.826 mmol) was added to a solution of 7-bromo-3,4-dichloroquinoline 1-oxide (220 mg, 0.751 mmol) in DCM (3129 μL). Meanwhile, triethylamine (523 μL, 3.76 mmol) was added to a separate suspension of oven-dried ammonium chloride (201 mg, 3.76 mmol) in DCM (6258 μL). The solution of tosylate was added dropwise to an ammonia solution. 80 mg of Ts-Cl was added to the mixture, and the reaction mixture was stirred overnight. An additional 100 mg of Ts-Cl was added. After approximately 45 minutes, a suspension prepared from 100 mg of ammonium chloride and 260 μL of triethylamine in 3 mL of DCM, which had been stirred for about 45 minutes, was added, and the reaction mixture was stirred overnight. An additional 100 mg of Ts-Cl was added, the mixture was stirred overnight, and then filtered. The filtrate was allowed to stand and then filtered again. The combined solid was washed with water and dried under reduced pressure to obtain 7-bromo-3,4-dichloroquinoline-2-amine (91 mg, 0.312 mmol, 42% yield).

[0446] [M+H] + = 292.8 and 294.6.

[0447] 1 ¹H NMR (400 MHz, methanol-d4) δ 7.89 (d, J=8.9 Hz, 1H), 7.77 (d, J=1.8 Hz, 1H), and 7.51–7.42 (m, 1H).

[0448] 3C. 3,4-Dichloro-7-(1-(tetrahydro-2H-pyran-2-yl)-1H-pyrazol-5-yl)quinoline-2-amine

[0449]

[0450] A mixture of 1-(tetrahydro-2H-pyran-2-yl)-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazole (108 mg, 0.390 mmol), 7-bromo-3,4-dichloroquinoline-2-amine (91 mg, 0.312 mmol), and PdCl2(dppf)-CH2Cl2 adduct (25.5 mg, 0.031 mmol) was placed in a pressure vial. The vial was placed under vacuum and refilled with nitrogen three times. Dioxane (1.6 mL) and tripotassium phosphate (2 M aqueous, 468 μL, 0.935 mmol) were added, nitrogen was bubbled through the solution, and the reaction mixture was heated to 100°C. The reaction mixture was cooled, diluted with water, and extracted three times with EtOAc. The organic layer was dried with sodium sulfate and concentrated. The residue was purified by ISCO silica gel chromatography (24 g column; CH2Cl2 / MeOH; 0-8% gradient) to obtain 3,4-dichloro-7-(1-(tetrahydro-2H-pyran-2-yl)-1H-pyrazole-5-yl)quinoline-2-amine (60 mg, 0.165 mmol, 53% yield).

[0451] LC / MS [M+H] + = 363.0 and 365.0.

[0452] 1 H NMR (400 MHz, methanol-d4) δ 8.11 (d, J=8.6 Hz, 1H), 7.77 (d, J=1.4 Hz, 1H), 7.63 (d, J=1.7 Hz, 1H), 7.52 (dd, J=8.5, 1.7 Hz, 1H), 6.53 (d, J=1.9 Hz, 1H), 5.33 (dd, J=10.2, 2.2 Hz, 1H), 4.15 - 4.08 (m, 1H), 3.71 - 3.63 (m, 1H), 2.54 - 2.41 (m, 1H), 2.07 - 1.99 (m, 1H), 1.87 - 1.79 (m, 1H), and 1.75 - 1.49 (m, 3H).

[0453] Example 3

[0454] Finnig base (0.036 mL, 0.206 mmol) was added to a solution of 3-aminopropan-1-ol (38.8 mg, 0.516 mmol) and 3,4-dichloro-7-(1-(tetrahydro-2H-pyran-2-yl)-1H-pyrazole-5-yl)quinoline-2-amine (25 mg, 0.069 mmol) in DMSO (0.5 mL). The reaction mixture was heated to 120°C. The reaction mixture was cooled, diluted with water, and extracted three times with EtOAc. The organic layer was concentrated. The residue was dissolved in 0.4 mL DCM and 0.2 mL TFA. After approximately 1 hour, the reaction mixture was concentrated and azeotropically mixed with DCM. The material was dissolved in MeOH, and K2CO3 was added. After approximately 2 hours, the reaction mixture was filtered, quenched with AcOH, and purified by reverse-phase LC / MS under the following conditions: Column: Exbridge C18, 200 mm x 19 mm, 5-μm particles; Mobile phase A: 5:95 acetonitrile:water, containing 10-mM ammonium acetate; Mobile phase B: 95:5 acetonitrile:water, containing 10-mM ammonium acetate; Gradient: hold at 1% B for 0 min, 1-41% B over 20 min, followed by hold at 100% B for 4 min; Flow rate: 20 mL / min; Column temperature: 25°C. The fractions containing the target product were combined and dried by centrifugal evaporation to obtain 3-((2-amino-3-chloro-7-(1H-pyrazole-3-yl)quinoline-4-yl)amino)propan-1-ol (10.2 mg).

[0455] Example 4. 3-((2-amino-8-fluoro-7-(1H-pyrazole-3-yl)quinoline-4-yl)amino)propan-1-ol

[0456]

[0457] 4A. 5-(((3-bromo-2-fluorophenyl)amino)methylene)-2,2-dimethyl-1,3-dioxane-4,6-dione

[0458]

[0459] A mixture of 3-bromo-2-fluoroaniline (1.15 g, 6.05 mmol) and 5-(methoxymethylene)-2,2-dimethyl-1,3-dioxane-4,6-dione (1.352 g, 7.26 mmol) in 5 mL of dioxane was heated at 120°C for 30 minutes. Subsequently, the reaction mixture was cooled to room temperature and diluted with 50 mL of diethyl ether. The solid was filtered and dried to obtain 5-(((3-bromo-2-fluorophenyl)amino)methylene)-2,2-dimethyl-1,3-dioxane-4,6-dione (1.76 g, 5.1 mmol, 85% yield).

[0460] 1 H NMR (400 MHz, DMSO-d6) δ 11.33 (br d, J=13.4 Hz, 1H), 8.67 (d, J=13.8 Hz, 1H), 7.91 - 7.81 (m, 1H), 7.59 (ddd, J=8.0, 6.6, 1.4 Hz, 1H), 7.25 (td, J=8.2, 1.4 Hz, 1H), and 1.70 (s, 6H).

[0461] 4B. 7-Bromo-8-fluoroquinoline-4-ol

[0462]

[0463] A solution of 5-(((3-bromo-2-fluorophenyl)amino)methylene)-2,2-dimethyl-1,3-dioxane-4,6-dione (1050 mg, 3.05 mmol) in diphenyl ether (7 mL) was heated at 240°C for 5 minutes. The reaction mixture was cooled to room temperature and diluted with 50 mL of diethyl ether. The solid was collected by vacuum filtration and dried to obtain 7-bromo-8-fluoroquinoline-4-ol (0.598 g, 2.5 mmol, 81% yield).

[0464] 1H NMR (400 MHz, DMSO-d6) δ 11.98 (br s, 1H), 7.87 (dd, J=7.2, 6.1 Hz, 1H), 7.82 (dd, J=8.7, 1.1 Hz, 1H), 7.54 (dd, J=8.7, 6.3 Hz, 1H), and 6.12 (d, J=7.4 Hz, 1H).

[0465] 4C. 7-Bromo-4-chloro-8-fluoroquinoline

[0466]

[0467] POCl3 (0.616 ml, 6.61 mmol) was added to a suspension of 7-bromo-8-fluoroquinoline-4-ol (800 mg, 3.31 mmol) in toluene (7 mL). The reaction mixture was heated at 100°C for 1 hour. The cooled reaction mixture was poured over ice and then distributed between DCM and saturated sodium carbonate. The organic layer was dried over sodium sulfate, concentrated under reduced pressure, and the residue was purified by ISCO silica gel chromatography (40 g column; hexane / ethyl acetate; 0-100% gradient) to obtain 7-bromo-4-chloro-8-fluoroquinoline (821 mg, 3.10 mmol, 95% yield).

[0468] 1 H NMR (400 MHz, DMSO-d6) δ 11.98 (br s, 1H), 7.87 (dd, J=7.2, 6.1 Hz, 1H), 7.82 (dd, J=8.7, 1.1 Hz, 1H), 7.54 (dd, J=8.7, 6.3 Hz, 1H), and 6.12 (d, J=7.4 Hz, 1H).

[0469] 4D. 7-Bromo-4-chloro-8-fluoroquinoline-1-oxide

[0470]

[0471] mCPBA (1.78 g, 7.22 mmol) was added to a solution of 7-bromo-4-chloro-8-fluoroquinoline (0.470 g, 1.804 mmol) in DCM (40.0 ml). The reaction mixture was stirred overnight and then quenched with a saturated sodium thiosulfate solution. The reaction mixture was stirred for 0.5 hours and then saturated aqueous sodium bicarbonate was added. The reaction mixture was extracted twice with DCM, the organic layer was washed with brine, dried with sodium sulfate, and concentrated to obtain 7-bromo-4-chloro-8-fluoroquinoline-1-oxide (0.215 g, 0.779 mmol, quantitative yield), which was used in subsequent steps without any further purification.

[0472] 4E. 7-Bromo-4-chloro-8-fluoroquinoline-2-amine

[0473]

[0474] In a round-bottom flask, 7-bromo-4-chloro-8-fluoroquinoline-1-oxide (215 mg, 0.779 mmol) was suspended in DCM (20 mL). Ts-Cl (378 mg, 1.99 mmol) was added, and the mixture was stirred for 1 hour. In a second round-bottom flask, ammonium chloride (483 mg, 9.02 mmol) (dried overnight in an oven at 110°C) was suspended in DCM (20 mL). Triethylamine (1.26 mL, 9.02 mmol) was added, and the mixture was stirred for 1 hour. The contents of the first flask were added to the second flask, the reaction mixture was stirred overnight, filtered, and concentrated. The residue was purified by ISCO silica gel chromatography (40 g column; DCM / MeOH 0-30% gradient) to obtain 7-bromo-4-chloro-8-fluoroquinoline-2-amine (220 mg, 0.79 mmol, 44% yield).

[0475] 4F. 4-chloro-8-fluoro-7-(1-(tetrahydro-2H-pyran-2-yl)-1H-pyrazol-5-yl)quinoline-2-amine

[0476]

[0477] (1-(tetrahydro-2H-pyran-2-yl)-1H-pyrazole-5-yl)boronic acid (0.222 g, 0.799 mmol), 7-bromo-4-chloro-8-fluoroquinoline-2-amine (0.220 g, 0.799 mmol), and PdCl2(dppf)-DCM adduct (0.065 g, 0.080 mmol) were placed in a pressure vial. The vial was placed under vacuum and refilled with nitrogen three times. Dioxane (10 mL) and tripotassium phosphate (2 M aqueous, 1.2 ml, 2.4 mmol) were added, and nitrogen was bubbled through the solution. The reaction mixture was heated to 100°C for 2 hours. The reaction mixture was cooled to room temperature, diluted with 50 mL of DCM, dried over sodium sulfate, and concentrated. The residue was purified by ISCO silica gel chromatography (24 g column; hexane / ethyl acetate 0-100% gradient) to obtain 4-chloro-8-fluoro-7-(1-(tetrahydro-2H-pyran-2-yl)-1H-pyrazole-5-yl)quinoline-2-amine (0.154 g, 0.22 mmol, 56% yield).

[0478] Example 4

[0479] Finnig base (0.5 mL, 2.88 mmol) was added to a solution of 4-chloro-8-fluoro-7-(1-(tetrahydro-2H-pyran-2-yl)-1H-pyrazole-5-yl)quinoline-2-amine (100 mg, 0.288 mmol) and 3-aminopropan-1-ol (65 mg, 0.865 mmol) in DMSO (0.5 mL). The reaction mixture was heated overnight at 120°C. The reaction mixture was cooled to room temperature, and 4N HCl (2 mL, 8 mmol) in dioxane was added. After 20 minutes, the reaction mixture was concentrated, the residue was diluted with DMF (1 mL), filtered through a syringe filter, and the crude material was purified by reverse-phase LC / MS under the following conditions: Column: Exbridge C18, 200 mm x 19 mm, 5-μm particles; Mobile phase A: 5:95 acetonitrile:water, containing 10-mM ammonium acetate; Mobile phase B: 95:5 acetonitrile:water, containing 10-mM ammonium acetate; Gradient: hold at 0% B for 0 min, 0-40% B over 20 min, followed by hold at 100% B for 4 min; Flow rate: 20 mL / min; Column temperature: 25°C. Fractions containing the target product were combined and dried by centrifugation. The material was further purified by purification LC / MS under the following conditions: Column: Exbridge C18, 200 mm x 19 mm, 5-μm particles; Mobile phase A: 5:95 acetonitrile:water, containing 10 mM ammonium acetate; Mobile phase B: 95:5 acetonitrile:water, containing 10 mM ammonium acetate; Gradient: 0% B for 0 minutes, 0-40% B over 20 minutes, followed by 100% B for 4 minutes; flow rate: 20 mL / min; column temperature: 25℃. Fractions containing the target product were combined and dried by centrifugal evaporation to obtain 3-((2-amino-8-fluoro-7-(1H-pyrazole-5-yl)quinoline-4-yl)amino)propan-1-ol (3.7 mg, 7.1%).

[0480] Example 5. (1S,3S)-3-((2-amino-5-methyl-7-(1H-pyrazole-3-yl)quinoline-4-yl)amino)cyclopentan-1-ol

[0481]

[0482] 5A. 7-Bromo-5-methylquinoline-4-ol

[0483]

[0484] A solution of 3-bromo-5-methylaniline (2.0 g, 10.75 mmol) and 5-(methoxymethylene)-2,2-dimethyl-1,3-dioxane-4,6-dione (2.00 g, 10.8 mmol) was stirred in diphenyl ether (7.2 mL) at 120°C for 30 minutes, followed by stirring with a vent needle at 240°C for 30 minutes. The reaction mixture was cooled to room temperature, hexane was added to precipitate the target product, filtered, and rinsed with hexane. The solid was dried to obtain 7-bromo-5-methylquinoline-4-ol (2.3 g, 90% yield) and its positional isomer, 5-bromo-4-chloro-7-methylquinoline-2-amine, as a yellowish-brown solid.

[0485] LC / MS [M+H] + = 237.9.

[0486] 5B. 7-Bromo-4-chloro-5-methylquinoline

[0487]

[0488] POCl3 (1.80 mL, 19.3 mmol) was added to a solution of 7-bromo-5-methylquinoline-4-ol (2.3 g, 9.66 mmol) in toluene (10.9 mL). The reaction mixture was stirred at 100°C for 1 hour, cooled to room temperature, and transferred to ice water in a beaker. While melting the ice, the mixture was diluted with DCM. The cold two-phase mixture was treated with 1.5 M dibasic potassium phosphate until the aqueous phase reached a pH of 7. The mixture was extracted with DCM (3x), the combined organic layer was washed with brine, and dried on MgSO4. It was concentrated under reduced pressure to obtain 7-bromo-4-chloro-5-methylquinoline (2.4 g, 98%) and its positional isomers as a dark brown oil.

[0489] LC / MS [M+H] + = 255.9.

[0490] 1 ¹H NMR (400 MHz, chloroform-d) δ 8.72–8.64 (m, 1H), 8.22–8.14 (m, 1H), 7.06–6.96 (m, 2H), and 3.04–3.01 (m, 3H).

[0491] 5C. 7-Bromo-4-chloro-5-methylquinoline-2-amine

[0492]

[0493] mCPBA (4.88 g, 28.3 mmol) was added to a solution of 7-bromo-4-chloro-5-methylquinoline (2.42 g, 9.43 mmol) in DCM (63 mL), and the reaction mixture was stirred overnight, followed by quenching with saturated aqueous sodium thiosulfate (18.9 mL, 47.2 mmol) while stirring for 1 hour. The reaction mixture was diluted with ethyl acetate, washed with 1.5 M dibasic potassium phosphate, washed with water, washed with brine, and dried over MgSO4. The amber oil was obtained by concentrating under reduced pressure. Ts-Cl (1.978 g, 10.37 mmol) was added to N-oxide (2.57 g, 9.43 mmol) in DCM (94 mL) at room temperature under nitrogen. The resulting homogeneous mixture was stirred for 1 hour. After filling with oven-dried ammonium chloride (2.52 g, 47.2 mmol), DCM (94 mL), and triethylamine (6.57 mL, 47.2 mmol), this solution was added via syringe to individual flasks that had been stirred for 30 minutes prior. Subsequently, the reaction mixture was stirred overnight, filtered, the filtrate concentrated, and the target product was purified by separating it from its positional isomer. Purification was carried out using a reverse-phase purification ISCO (150 g C18 column) with a gradient of 15-100% B (water / MeOH) to obtain 7-bromo-4-chloro-5-methylquinoline-2-amine (0.17 g, 15% yield) as a grayish-white solid.

[0494] LC / MS [M+H] + = 271.1.

[0495] 1 H NMR (500 MHz, DMSO-d6) δ 7.65 (s, 1H), 7.42 (br s, 1H), 7.10 - 6.99 (m, 1H), and 2.87 - 2.83 (m, 3H).

[0496] 5D. (1S,3S)-3-(2-amino-7-bromo-5-methylquinoline-4-ylamino)cyclopentanol

[0497]

[0498] To a solution of 7-bromo-4-chloro-5-methylquinoline-2-amine (0.030 g, 0.110 mmol) in NMP (0.55 ml), Finnig base (0.193 ml, 1.105 mmol) and (1S,3S)-3-aminocyclopentan-1-ol (0.034 g, 0.331 mmol) were added. Subsequently, the reaction mixture was stirred overnight at 170°C, then diluted with ethyl acetate and washed with water. The aqueous phase was extracted with ethyl acetate (3x), the combined organic layer was washed with saturated aqueous ammonium chloride, washed with water, washed with brine, and dried on MgSO4. (1S,3S)-3-(2-amino-7-bromo-5-methylquinoline-4-ylamino)cyclopentanol (17 mg, 27% yield) was obtained as an amber oil by concentrating under reduced pressure.

[0499] LC / MS [M+H] + = 336.2.

[0500] Example 5

[0501] A mixture of (1S,3S)-3-((2-amino-7-bromo-5-methylquinoline-4-yl)amino)cyclopentan-1-ol (0.0173 g, 0.051 mmol), 3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazole (0.013 g, 0.067 mmol), and tripotassium phosphate (0.077 ml, 0.154 mmol) was degassed (3x) in dioxane (0.322 mL). 1,1'-bis(di-tert-butylphosphino)ferrocene palladium dichloride (3.35 mg, 5.15 μmol) was added to the mixture, the mixture was degassed again (3x), and then stirred overnight at 80°C. Dioxane was removed under reduced pressure, the crude material dissolved in DMF was filtered, and purified by reverse-phase LC / MS under the following conditions: Column: Exbridge C18, 200 mm x 19 mm, 5-μm particles; Mobile phase A: 5:95 acetonitrile:water, containing 10-mM ammonium acetate; Mobile phase B: 95:5 acetonitrile:water, containing 10-mM ammonium acetate; Gradient: 0-min held at 2% B, 2-42% B over 25 minutes, followed by 4-min held at 100% B; Flow rate: 20 mL / min to obtain (1S,3S)-3-(2-amino-5-methyl-7-(1H-pyrazole-3-yl)quinoline-4-ylamino)cyclopentanol (1.0 mg, 6% yield).

[0502] Example 6. N4-((1H-pyrazole-3-yl)methyl)-5-methyl-7-(1H-pyrazole-3-yl)quinoline-2,4-diamine

[0503]

[0504] A mixture of N4-((1H-pyrazole-3-yl)methyl)-7-bromo-5-methylquinoline-2,4-diamine (0.0125 g, 0.038 mmol, prepared as outlined in the preparation of Example 5), 3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazole (9.49 mg, 0.049 mmol), and tripotassium phosphate (0.056 mL, 0.113 mmol) was degassed (3x) in dioxane (0.24 mL). 1,1'-bis(di-tert-butylphosphino)ferrocene palladium dichloride (2.45 mg, 3.76 μmol) was added to the mixture, the mixture was degassed again (3x), and stirred overnight at 80°C. Dioxane was removed under reduced pressure, the crude material was dissolved in DMF, filtered, and purified by LC / MS for purification under the following conditions: Column: Exbridge C18, 200 mm x 19 mm, 5-μm particles; Mobile phase A: 5:95 acetonitrile:water, containing 10-mM ammonium acetate; Mobile phase B: 95:5 acetonitrile:water, containing 10-mM ammonium acetate; Gradient: 0-min hold at 0% B, 0-30% B over 25 minutes, followed by 4-min hold at 100% B; Flow rate: 20 mL / min to obtain N4-((1H-pyrazole-3-yl)methyl-7-(1H-pyrazole-3-yl)quinoline-2,4-diamine (2.2 mg, 18% yield).

[0505] Example 7. 3-((2-amino-5-fluoro-7-(1H-pyrazole-3-yl)quinoline-4-yl)amino)propan-1-ol

[0506]

[0507] 7A. 5-(((3-bromo-5-fluorophenyl)amino)methylene)-2,2-dimethyl-1,3-dioxane-4,6-dione

[0508]

[0509] A heterogeneous mixture of 5-(methoxymethylene)-2,2-dimethyl-1,3-dioxane-4,6-dione (1.959 g, 10.53 mmol), 5-(((3-bromo-5-fluorophenyl)amino)methylene)-2,2-dimethyl-1,3-dioxane-4,6-dione, and diphenyl ether (7.02 mL) was immersed in an oil bath at 120°C and stirred for 20 minutes (the reaction mixture became homogeneous). The reaction mixture was used in subsequent steps without any additional post-treatment or purification.

[0510] 7B. 7-Bromo-5-fluoroquinoline-4-ol

[0511]

[0512] Reaction mixture form step A (40 mL screw-top vial) was heated at 240°C for 15 minutes. In addition to the mixture, 3x similar reactants were rapidly cooled and added to diethyl ether (300 mL) while stirring. The precipitates produced from all four reactions were collected by vacuum filtration to obtain 3.53 g of a 1:1 mixture, 7-bromo-5-fluoroquinoline-4-ol and 5-bromo-7-fluoroquinoline-4-ol, as yellowish-brown solids, which were used without further purification.

[0513] LC / MS [M+H] + = 242.0 and 244.0.

[0514] 7C. 7-Bromo-4-chloro-5-fluoroquinoline

[0515]

[0516] A mixture of 7-bromo-5-fluoroquinoline-4-ol (3.53 g, 14.58 mmol) (a mixture of two positional isomers) and POCl3 (0.832 mL, 8.93 mmol) in toluene (16.20 mL) was heated in an oil bath at 100°C for 60 minutes. The reaction mixture was concentrated under reduced pressure, the residue was diluted with dichloromethane, washed with a saturated aqueous solution of sodium bicarbonate, and dried over anhydrous sodium sulfate. The mixture was concentrated under reduced pressure and subsequently purified by ISCO flash silica gel chromatography (220 g column; 0-50% ethyl acetate in hexane) to obtain a mixture (2.16 g) of 7-bromo-4-chloro-5-fluoroquinoline and 5-bromo-4-chloro-7-fluoroquinoline as a pale yellow solid.

[0517] LC / MS [M+H] + = 260.0 and 262.0.

[0518] 7D. 7-Bromo-4-chloro-5-fluoroquinoline 1-oxide

[0519]

[0520] mCPBA (4.3 g, 24.9 mmol) was added to a solution of 7-bromo-4-chloro-5-fluoroquinoline (2.16 g, 8.29 mmol) (a mixture of two positional isomers) in dichloromethane (55 mL) at room temperature. The reaction mixture was stirred overnight at room temperature. The reaction mixture was quenched with a saturated aqueous sodium thiosulfate solution, slowly added, and stirred for 30 minutes. The mixture was diluted with dichloromethane and washed with saturated aqueous sodium bicarbonate (2x). The organic layer was collected, and the aqueous layer was extracted with dichloromethane (2x). The combined organic layer was dried and concentrated over anhydrous sodium sulfate to obtain a mixture of 7-bromo-4-chloro-5-fluoroquinoline 1-oxide and 5-bromo-4-chloro-7-fluoroquinoline 1-oxide (2.00 g, 87% yield) as a pale yellow solid.

[0521] LC / MS [M+H] + = 276.0 and 278.0 (two close peaks in LC).

[0522] 7E. 7-Bromo-4-chloro-5-fluoroquinoline-2-amine

[0523]

[0524] Tosyl-Cl (1.52 g, 7.96 mmol) was added to a solution (100 mL round-bottom flask) of 7-bromo-4-chloro-5-fluoroquinoline 1-oxide (2.00 g, 7.23 mmol) (a mixture of two positional isomers) in dichloromethane (20 mL). The resulting homogeneous mixture was stirred at room temperature for 30 minutes. In a second flask (100 mL round-bottom) containing a solution of triethylamine (5.04 mL, 36.2 mmol) and dichloromethane (50 mL), ammonium chloride (1.94 g, 36.2 mmol) (oven-dried) was added. The first reaction mixture was added to the second flask, and the resulting reaction mixture was stirred overnight at room temperature. The reaction mixture was filtered, concentrated, and the resulting residue (~2.0 g) was subjected to: 1.) reverse-phase iscochromatography (1.43 g); and 2.) A mixture of 7-bromo-4-chloro-5-fluoroquinoline-2-amine and 5-bromo-4-chloro-7-fluoroquinoline-2-amine (1.05 g, 53%) was obtained as a grayish-white solid by purification by ISCO flash silica gel chromatography (80 g column; 0-20% methanol in dichloromethane).

[0525] LC / MS [M+H] + = 275.0 and 277.0 (two close peaks in LC).

[0526] 7F. 3-((2-amino-7-bromo-5-fluoroquinoline-4-yl)amino)propan-1-ol

[0527]

[0528] 2. In a screw-stopped vial of Dram, a mixture of 7-bromo-4-chloro-5-fluoroquinoline-2-amine / 5-bromo-4-chloro-7-fluoroquinoline-2-amine (0.120 g, 0.436 mmol), 3-aminopropan-1-ol (0.067 mL, 0.871 mmol), Finnig base (0.38 mL, 2.18 mmol), and DMSO (0.5 mL) was heated overnight in an oil bath at 120°C. The reaction mixture was diluted with ethyl acetate, washed with 10% aqueous lithium chloride (2x), and washed with brine. The organic layer was collected, and the aqueous layer was sequentially extracted with ethyl acetate (2x). The combined organic layer was dried over anhydrous sodium sulfate. After concentration under reduced pressure, the product was subsequently purified by ISCO silica gel chromatography (12 g column in dichloromethane; 0-30% methanol) to obtain 3-((2-amino-7-bromo-5-fluoroquinoline-4-yl)amino)propan-1-ol (0.012 mg, 0.036 μmol, 0.083% yield) as a pale yellow solid (the first of two overlapping peaks). A significant mixed fraction and undesirable products were retained for further purification.

[0529] LC / MS [M+H] + = 314.0 and 316.0.

[0530] 1 ¹H NMR (400 MHz, methanol-d4) δ 7.46 (s, 1H), 7.24 (m, 1H), 5.85 (s, 1H), 3.77 (t, J=5.8 Hz, 2H), 3.48–3.42 (m, 2H), and 1.99–1.94 (m, 2H).

[0531] Example 7

[0532] A mixture of 3-((2-amino-7-bromo-5-fluoroquinoline-4-yl)amino)propan-1-ol (0.0195 g, 0.062 mmol), 3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazole (0.024 g, 0.124 mmol), and tripotassium phosphate (2 M in water) (0.093 mL, 0.186 mmol) was degassed (3x; vacuum / nitrogen) in dioxane (1.2 mL). 1,1'-bis(di-tert-butylphosphino)ferrocene palladium dichloride (5.07 mg, 6.21 μmol) was added to the mixture, and the mixture was degassed (3x; vacuum / nitrogen). The reaction mixture was immersed in an oil bath at 85°C and stirred overnight. The reaction mixture was diluted with ethyl acetate, washed with water, and washed with brine. The organic layer was collected, and the aqueous layer was sequentially extracted with ethyl acetate (3x). The combined organic layer was dried over anhydrous sodium sulfate and concentrated under reduced pressure. The residue was purified by ISCO silica gel chromatography (4 g column; 0-20% methanol in dichloromethane) to obtain 3-((2-amino-5-fluoro-7-(1H-pyrazole-3-yl)quinoline-4-yl)amino)propan-1-ol (8.0 mg, 0.026 mmol, 42% yield) as a yellowish-brown solid. Additional product was isolated when the column was run up to 30% methanol in dichloromethane, but the fraction was not included.

[0533] Example 8. (1S,3S)-3-((2-amino-5-fluoro-7-(1H-pyrazole-3-yl)quinoline-4-yl)amino)cyclopentan-1-ol

[0534]

[0535] 8A. (1S,3S)-3-((2-amino-7-bromo-5-fluoroquinoline-4-yl)amino)cyclopentan-1-ol

[0536]

[0537] 2. In a screw-stopped vial of Dram, a mixture of 7-bromo-4-chloro-5-fluoroquinoline-2-amine / 5-bromo-4-chloro-7-fluoroquinoline-2-amine (7E, 0.180 g, 0.653 mmol), (1S,3S)-3-aminocyclopentan-1-ol, HCl (0.180 g, 1.31 mmol), Finnig base (0.571 mL, 3.27 mmol), and DMSO (0.8 mL) was heated overnight in an oil bath at 120°C. The reaction mixture was diluted with ethyl acetate, washed with 10% aqueous lithium chloride (2x), and washed with brine. The organic layer was collected, and the aqueous layer was sequentially extracted with ethyl acetate (2x). The combined organic layer was dried over anhydrous sodium sulfate. Concentrated under reduced pressure and subsequently purified by ISCO silica gel chromatography (12 g in dichloromethane; 0-30% methanol) to obtain (1S,3S)-3-((2-amino-7-bromo-5-fluoroquinoline-4-yl)amino)cyclopentan-1-ol as a pale yellow solid (the first of two overlapping peaks). Significant mixed fractions and undesirable products were concentrated and retained for further purification.

[0538] LC / MS [M+H] + = 340.2 and 342.2.

[0539] Example 8

[0540] A mixture of (1S,3S)-3-((2-amino-7-bromo-5-fluoroquinoline-4-yl)amino)cyclopentan-1-ol (0.019 g, 0.056 mmol), 3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazole (0.022 g, 0.112 mmol), and potassium carbonate (2 M in water) (0.084 mL, 0.168 mmol) was degassed (3x; vacuum / nitrogen) in dioxane (1.2 mL). 1,1'-bis(di-tert-butylphosphino)ferrocene palladium dichloride (4.6 mg, 5.59 μmol) was added to the mixture, and the mixture was degassed (3x; vacuum / nitrogen). The reaction mixture was immersed in an oil bath at 85°C and stirred overnight. The reaction mixture was diluted with ethyl acetate, washed with water, and washed with brine. The organic layer was collected, and the aqueous layer was sequentially extracted with ethyl acetate (3x). The combined organic layer was dried over anhydrous sodium sulfate and concentrated under reduced pressure. The residue was purified by ISCO silica gel chromatography (4 g column in dichloromethane; 0-20% methanol) to obtain (1S,3S)-3-((2-amino-5-fluoro-7-(1H-pyrazole-3-yl)quinoline-4-yl)amino)cyclopentan-1-ol (6.0 mg, 0.017 mmol, 31.2% yield) as a yellowish-brown solid. Additional product was isolated when the column was run up to 30% methanol in dichloromethane, but the fraction was not included.

[0541] Example 9. N4-((1H-pyrazole-3-yl)methyl)-5-fluoro-7-(1H-pyrazole-3-yl)quinoline-2,4-diamine

[0542]

[0543] 9A. N4-((1H-pyrazole-3-yl)methyl)-7-bromo-5-fluoroquinoline-2,4-diamine

[0544]

[0545] 2. In a screw-stopped vial of Dram, 7-bromo-4-chloro-5-fluoroquinoline-2-amine / 5-bromo-4-chloro-7-fluoroquinoline-2-amine (7E, 0.150 g, 0.544 mmol), (1H-pyrazole-3-yl)methaneamine (0.106 g, 1.089 mmol) and a mixture of other positional isomers, Finnig base (0.48 mL, 2.72 mmol), and DMSO (0.6 mL) were heated overnight in an oil bath at 120°C. The reaction mixture was diluted with ethyl acetate, washed with 10% aqueous lithium chloride (2x), and washed with brine. The organic layer was collected, and the aqueous layer was sequentially extracted with ethyl acetate (2x). The combined organic layer was dried over anhydrous sodium sulfate. Concentrated under reduced pressure and then purified by ISCO silica gel chromatography (compounds pre-adsorbed onto silica gel) (12 g column; 0-30% methanol in dichloromethane) to obtain a mixture of N4-((1H-pyrazole-3-yl)methyl)-7-bromo-5-fluoroquinoline-2,4-diamine and N4-((1H-pyrazole-3-yl)methyl)-5-bromo-7-fluoroquinoline-2,4-diamine as a pale yellow solid (33 mg, 18% yield).

[0546] LC / MS [M+H] + = 314.0 and 316.0.

[0547] Example 9

[0548] A mixture of N4-((1H-pyrazole-3-yl)methyl)-7-bromo-5-fluoroquinoline-2,4-diamine (plus other positional isomers) (0.033 g, 0.098 mmol), 3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazole (0.038 g, 0.196 mmol), and potassium carbonate (2.0 M in water) (0.147 mL, 0.294 mmol) was degassed (3x; vacuum / nitrogen) in dioxane (1.5 mL). 1,1'-bis(di-tert-butylphosphino)ferrocene palladium dichloride (8.02 mg, 9.82 μmol) was added to the mixture, and the mixture was degassed (3x; vacuum / nitrogen). The reactants were immersed in an oil bath at 85°C and stirred overnight. Analysis of the reaction mixture indicated that only the target positional isomer reacted, while the untarget positional isomer remained intact. The reaction mixture was diluted with ethyl acetate, washed with water, and washed with brine. The organic layer was collected, and the aqueous layer was sequentially extracted with ethyl acetate (3x). The combined organic layer was dried over anhydrous sodium sulfate, concentrated under reduced pressure, and purified by reverse-phase LC / MS under the following conditions: Column: Exbridge C18, 200 mm x 19 mm, 5-μm particles; Mobile phase A: 5:95 acetonitrile:water, containing 10-mM ammonium acetate; Mobile phase B: 95:5 acetonitrile:water, containing 10-mM ammonium acetate; Gradient: 0% B for 0 minutes, 0-30% B over 23 minutes, followed by 100% B for 4 minutes; flow rate: 20 mL / min; column temperature: 25℃. Fractions containing the target product were combined and dried by centrifugal evaporation to obtain N4-((1H-pyrazole-3-yl)methyl)-5-fluoro-7-(1H-pyrazole-3-yl)quinoline-2,4-diamine (3.6 mg).

[0549] Example 10. (S)-5-fluoro-7-(1H-pyrazole-3-yl)-N4-(tetrahydrofuran-3-yl)quinoline-2,4-diamine

[0550]

[0551] 10A. (S)-7-bromo-5-fluoro-N4-(tetrahydrofuran-3-yl)quinoline-2,4-diamine

[0552]

[0553] In a screw-stopped vial of 2 Drams, a mixture of 7-bromo-4-chloro-5-fluoroquinoline-2-amine (mixture of two positional isomers) (0.050 g, 0.181 mmol), (S)-tetrahydrofuran-3-amine (0.024 g, 0.272 mmol), Finnig base (0.158 mL, 0.907 mmol), and DMSO (0.2 mL) was heated overnight in an oil bath at 120°C. The reaction mixture was diluted with ethyl acetate, washed with 10% aqueous lithium chloride (2x), and washed with brine. The organic layer was collected, and the aqueous layer was sequentially extracted with ethyl acetate (2x). The combined organic layer was dried over anhydrous sodium sulfate. After concentrating under reduced pressure, the mixture (12 mg) of (S)-7-bromo-5-fluoro-N4-(tetrahydrofuran-3-yl)quinoline-2,4-diamine and (S)-5-bromo-7-fluoro-N4-(tetrahydrofuran-3-yl)quinoline-2,4-diamine was subsequently purified by ISCO silica gel chromatography (12 g column; 0-30% methanol in dichloromethane) to obtain a pale yellow solid.

[0554] LC / MS [M+H] + = 326.1 and 328.1.

[0555] Example 10

[0556] A mixture of (S)-7-bromo-5-fluoro-N4-(tetrahydrofuran-3-yl)quinoline-2,4-diamine and (S)-5-bromo-7-fluoro-N4-(tetrahydrofuran-3-yl)quinoline-2,4-diamine (0.012 g, 0.037 mmol), 3-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazole (0.014 g, 0.074 mmol), and potassium carbonate (2 M in water, 0.055 mL, 0.110 mmol) was degassed (3x; vacuum / nitrogen). 1,1'-bis(di-tert-butylphosphino)ferrocene palladium dichloride (3.00 mg, 3.68 μmol) was added to the mixture, and the mixture was degassed (3x; vacuum / nitrogen). The reaction mixture was immersed in an oil bath at 100°C and stirred overnight. Analysis of the reaction mixture indicated that the homogeneous reaction was complete, forming products from both positional isomers. The reaction mixture was diluted with ethyl acetate, washed with water, and washed with brine. The organic layer was collected, and the aqueous layer was sequentially extracted with ethyl acetate (3x). The combined organic layer was dried over anhydrous sodium sulfate and concentrated under reduced pressure. The residue was purified by ISCO silica gel chromatography (4 g column; 0-30% methanol in dichloromethane) to obtain ((S)-5-fluoro-7-(1H-pyrazole-3-yl)-N4-(tetrahydrofuran-3-yl)quinoline-2,4-diamine (1.5 mg, 4.55 μmol, 12% yield) as a yellowish-brown solid. Of the two overlapping peaks, the eluted first peak corresponded to the target product.

[0557] Example 11

[0558] N4-((1H-pyrazole-5-yl)methyl)-8-fluoro-7-(1H-pyrazole-5-yl)quinoline-2,4-diamine

[0559]

[0560] 11A. Methyl 2-amino-4-bromo-3-fluorobenzoate

[0561]

[0562] TMS-diazomethane (2.14 mL, 4.27 mmol, 2.0 M in hexane) was added to a solution of 2-amino-4-bromo-3-fluorobenzoic acid (500 mg, 2.14 mmol) in dichloromethane (5 mL) and methanol (5 mL). The resulting mixture was stirred at room temperature for 5 hours. The reaction mixture was concentrated to obtain the product as a gray solid (510 mg, 96% yield).

[0563] LC / MS [M+H] + = 248.0.

[0564] 1 ¹H NMR (400 MHz, methanol-d4) δ 7.59–7.48 (m, 1H), 6.82–6.76 (m, 1H), and 3.95–3.85 (m, 3H).

[0565] 11B. 2-amino-7-bromo-8-fluoroquinoline-4-ol

[0566]

[0567] n-butyllithium (0.825 mL, 9.07 mmol, 11.0 M in hexane) was added to a stirred solution of acetonitrile (0.948 mL, 18.14 mmol) in tetrahydrofuran (15 mL) at -78°C. The resulting mixture was stirred at -78°C for 1 hour. Subsequently, methyl 2-amino-4-bromo-3-fluorobenzoate (450 mg, 1.81 mmol) in 30 mL of tetrahydrofuran was added dropwise over 15 minutes, and the reaction mixture was continued at -78°C for 30 minutes. After heating to room temperature, the reaction mixture was stirred at room temperature for an additional 24 hours. A saturated aqueous solution of ammonium chloride was added, and the precipitate was collected by vacuum filtration to obtain the product as a white solid (300 mg, 1.17 mmol, 64% yield).

[0568] LC / MS [M+H] + = 258.9.

[0569] 11C. 2-amino-8-fluoro-7-(1H-pyrazol-5-yl)quinoline-4-ol

[0570]

[0571] A solution of 2-amino-7-bromo-8-fluoroquinoline-4-ol (50 mg, 0.195 mmol) in dioxane (2 mL) was added to a 10 mL reaction vial that had been purged and maintained under an inert atmosphere of nitrogen. To the solution, tribasic potassium phosphate (0.292 mL, 0.584 mmol, 2 M in water), 5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazole (45.3 mg, 0.233 mmol), and PdCl2(dppf) (7.12 mg, 9.73 μmol) were added. The resulting solution was stirred at 100°C for 13 hours. The reaction mixture was purified by reverse-phase HPLC to obtain 2-amino-8-fluoro-7-(1H-pyrazole-5-yl)quinoline-4-ol (35 mg, 0.137 mmol, 70% yield) as a white solid.

[0572] LC / MS [M+H] + = 245.1.

[0573] 1 ¹H NMR (400 MHz, methanol-d4) δ 8.05–8.01 (m, 1H), 7.99–7.96 (m, 1H), 7.87–7.81 (m, 1H), 6.98–6.88 (m, 1H), and 6.40–6.34 (m, 1H).

[0574] 11D. 4-chloro-8-fluoro-7-(1H-pyrazol-5-yl)quinoline-2-amine

[0575]

[0576] A solution of 2-amino-8-fluoro-7-(1H-pyrazole-5-yl)quinoline-4-ol (50 mg, 0.205 mmol) in POCl3 (954 μl, 10.24 mmol) was heated at 100°C for 12 hours. The reaction mixture was evaporated under reduced pressure. It was purified by reverse-phase HPLC to obtain 4-chloro-8-fluoro-7-(1H-pyrazole-5-yl)quinoline-2-amine (35 mg, 0.133 mmol, 65% yield).

[0577] LC / MS [M+H] + = 263.1.

[0578] 1 ¹H NMR (400 MHz, methanol-d4) δ 8.07–8.03 (m, 1H), 7.96–7.91 (m, 1H), 7.88–7.83 (m, 1H), 7.33–7.28 (m, 1H), and 7.00–6.93 (m, 1H).

[0579] Example 11

[0580] To a solution of 4-chloro-8-fluoro-7-(1H-pyrazole-5-yl)quinoline-2-amine (10 mg, 0.038 mmol) in NMP (1 mL), Finnig base (0.033 mL, 0.190 mmol) and (1H-pyrazole-5-yl)methaneamine (7.39 mg, 0.076 mmol) were added. The resulting mixture was heated at 150°C for 12 hours. The reaction mixture was purified by reverse-phase HPLC to obtain N4-((1H-pyrazole-5-yl)methyl)-8-fluoro-7-(1H-pyrazole-5-yl)quinoline-2,4-diamine (5 mg, 0.015 mmol, 39% yield).

[0581] Example 12. 3-((2-amino-3-methyl-7-(1H-pyrazole-5-yl)quinoline-4-yl)amino)propan-1-ol

[0582]

[0583] 12A. 2-amino-7-bromo-3-methylquinoline-4-ol

[0584]

[0585] n-butyllithium (0.988 mL, 10.87 mmol, 11.0 M in hexane) was added to a stirred solution of propionitrile (1.551 mL, 21.73 mmol) in tetrahydrofuran (18 mL) at -78°C. The resulting mixture was stirred at -78°C for 1 hour. Subsequently, methyl 2-amino-4-bromobenzoate (500 mg, 2.173 mmol) in 3 mL of tetrahydrofuran was added dropwise over 15 minutes. The reaction mixture was stirred at -78°C for 30 minutes, heated to room temperature, and stirred at room temperature for 48 hours. A saturated aqueous solution of ammonium chloride was added, the resulting precipitate was collected by vacuum filtration and dried to obtain 2-amino-7-bromo-3-methylquinoline-4-ol (300 mg, 1.19 mmol, 55% yield) as a white solid.

[0586] LC / MS [M+H] + = 254.9.

[0587] 1 ¹H NMR (400 MHz, methanol-d4) δ 8.07–7.95 (m, 1H), 7.81–7.71 (m, 1H), 7.62–7.49 (m, 1H), and 2.23–2.12 (m, 3H).

[0588] 12B. 2-amino-3-methyl-7-(1H-pyrazole-5-yl)quinoline-4-ol

[0589]

[0590] A solution of 2-amino-7-bromo-3-methylquinoline-4-ol (100 mg, 0.395 mmol) in dioxane (3 mL) was added to a 10 mL reaction vial that had been purged and maintained under an inert atmosphere of nitrogen. To the solution, 2 M tribasic potassium phosphate (0.593 mL, 1.19 mmol), 5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1H-pyrazole (92 mg, 0.474 mmol), and PdCl2(dppf) (14.5 mg, 0.020 mmol) in water were added. The resulting solution was stirred at 100°C for 13 hours. The reaction mixture was purified by reverse-phase HPLC to obtain 2-amino-3-methyl-7-(1H-pyrazole-5-yl)quinoline-4-ol (35 mg, 0.146 mmol, 37% yield) as a white solid.

[0591] LC / MS [M+H] + = 241.1.

[0592] 12C. 4-chloro-3-methyl-7-(1H-pyrazol-5-yl)quinoline-2-amine

[0593]

[0594] A solution of 2-amino-3-methyl-7-(1H-pyrazole-5-yl)quinoline-4-ol (30 mg, 0.125 mmol) in POCl3 (349 μl, 3.75 mmol) was heated at 100°C for 12 hours. The reaction mixture was concentrated under reduced pressure and dried to obtain 4-chloro-3-methyl-7-(1H-pyrazole-5-yl)quinoline-2-amine (30 mg, 0.116 mmol, 93% yield).

[0595] LC / MS [M+H] + = 259.0.

[0596] Example 12

[0597] To a solution of 4-chloro-3-methyl-7-(1H-pyrazole-5-yl)quinoline-2-amine (15 mg, 0.058 mmol) in NMP (1 mL), Finnig base (0.051 mL, 0.290 mmol) and 3-aminopropan-1-ol (8.71 mg, 0.116 mmol) were added. The resulting mixture was heated at 150°C for 12 hours and then purified by reverse-phase HPLC to obtain 3-((2-amino-3-methyl-7-(1H-pyrazole-5-yl)quinoline-4-yl)amino)propan-1-ol (12 mg, 0.040 mmol, 69.6% yield).

[0598] Example 13. N4-((1H-pyrazole-5-yl)methyl)-3-methyl-7-(1H-pyrazole-5-yl)quinoline-2,4-diamine

[0599]

[0600] The target compound was prepared in a manner similar to that used in the preparation of Example 12.

[0601] Example 14. 2-((2-amino-3-cyclopentyl-7-(1H-pyrazole-5-yl)quinoline-4-yl)amino)ethanol-1-ol

[0602]

[0603] 4-chloro-7-(1-(tetrahydro-2H-pyran-2-yl)-1H-pyrazole-5-yl)quinoline-2-amine (150 mg, 0.456 mmol)) and potassium cyclopentyl-trifluoroborate (80 mg, 0.456 mmol) were added to a reaction vial in acetic acid-water (1:1, 5 mL). TFA (35.1 μl, 0.456 mmol) was added, and the reaction mixture was stirred until homogeneous. Subsequently, manganese (III) acetate dihydrate (306 mg, 1.141 mmol) was added. The reaction mixture was sealed and heated overnight at 50°C. The cooled reaction mixture was filtered to obtain 111 mg of crude 4-chloro-3-cyclopentyl-7-(1H-pyrazole-5-yl)quinoline-2-amine. A portion of this substance (30 mg, 0.096 mmol) was dissolved in NMP (0.25 mL) in a reaction vial. Ethanolamine (29 μL, 0.480 mmol) was added. The vial was flushed with nitrogen, sealed, and heated to 160 °C. After reacting overnight, the cooled reaction was diluted with methanol and filtered. The crude substance was purified by purification LC / MS under the following conditions: Column: Exbridge C18, 200 mm x 19 mm, 5-μm particles; Mobile phase A: 5:95 Acetonitrile:water, containing ammonium acetate; Mobile phase B: 95:5 Acetonitrile:water, containing ammonium acetate; Gradient: 0-min hold at 5% B, 5-45% B over 20 minutes, followed by 0-min hold at 100% B; Flow rate: 20 mL / min; Column temperature: 25℃. Fraction collection was initiated by MS and UV signals. Fractions containing the target product were combined and dried by centrifugal evaporation to obtain 2-((2-amino-3-cyclopentyl-7-(1H-pyrazole-5-yl)quinoline-4-yl)amino)ethanol-1-ol (8.8 mg, 0.026 mmol).

[0604] Example 15. 3-Cyclopentyl-7-(1H-pyrazole-5-yl)-N4-(pyridine-2-ylmethyl)quinoline-2,4-diamine

[0605]

[0606] Example 15 was prepared in a manner similar to that described for Example 14.

[0607] Example 16. 3-((2-amino-3-cyclopentyl-7-(1H-pyrazole-5-yl)quinoline-4-yl)amino)propan-1-ol

[0608]

[0609] 4-chloro-3-cyclopentyl-7-(1H-pyrazole-5-yl)quinoline-2-amine (50 mg, 0.160 mmol) was added to a reaction vial in NMP (250 μL). 3-amino-1-propanol (61.1 μL, 0.799 mmol) was added. The vial was flushed with nitrogen, sealed, and heated to 160°C. After stirring overnight, the cooled reaction mixture was diluted with methanol, filtered, and purified by RP-HPLC (methanol-water gradient +0.1% TFA). The product containing the fraction was treated with a sodium bicarbonate solution. The solvent was evaporated until the sample became mostly water. The product was separated as a sticky oil. The sample was sonicated, and the liquid was carefully decanted. Water was added, and the process was repeated. The residue was dissolved in ethanol and evaporated. The sample was pumped down to obtain 3-((2-amino-3-cyclopentyl-7-(1H-pyrazole-5-yl)quinoline-4-yl)amino)propan-1-ol (18.5 mg, 0.051 mmol, 32.2% yield).

[0610]

[0611]

[0612]

[0613]

[0614] Many embodiments of the present invention have been described. However, it will be understood that various modifications may be made without departing from the spirit and scope of the invention. Accordingly, other embodiments are also within the scope of the following claims.

Claims

Claim 1 Compounds of chemical formulas (I), (II), or (III) or their stereoisomers, tautomers, or pharmaceutically acceptable salts. Here, W is independently -NH-R 6a or -NH-YR 6 And;Y is independently C 1-3 It is an alkylene and;R 1 and R 3 is H and;R 1a is a halogen and;R 2 is a heteroaryl containing five ring atoms independently, where two ring atoms are each independently selected from N and NH;R 4a is independently halogen and C 1-4 Selected from alkyl; R 4 is independently H or R 4a Igo;R 6 -OR independently a and R 6a Selected from;R 6a is a heteroaryl comprising 5 to 6 ring atoms independently (wherein 1 to 2 ring atoms are independently N and N(R f Selected from ); 0 to 1 R g C substituted with 5-6 Selected from cycloalkyl; and heterocyclyl comprising 5 to 6 ring atoms (wherein one ring atom is O); and R a H and C independently 1-4 Selected from alkyl; R g is OH and;R 7 is H and;R 7a is a halogen, C 1-4 alkyl, or C 3-6 It is cycloalkyl and;R f is H. Claim 2 In paragraph 1, W independently -NH-R 6a or -NH-YR 6 and;Y independently C 1-3 It is an alkylene and;R 1 and R 3 This is H and;R 7 This is H and;R 4 In each case, is independently selected from H, F, and CH3; R 1a is independently F or Cl and R 4a is independently selected from F, Cl, and CH3; R 6 These independently consist of OH, OCH3, and R 6a Selected from;R 6a α independently pirazolyl, cyclopentyl substituted with OH; and A compound selected from. Claim 3 In paragraph 1, Or any one compound selected from salts permitted by his constraints. Claim 4 A pharmaceutical composition for use in the treatment of cancer, comprising a compound according to claim 1 or a pharmaceutically acceptable salt thereof and one or more pharmaceutically acceptable excipients. Claim 5 In paragraph 4, cancer is acute myeloid leukemia, adrenocortical carcinoma, Kaposi sarcoma, lymphoma, anal cancer, appendiceal cancer, teratoid / rhabdomyoplastic tumor, basal cell carcinoma, cholangiocarcinoma, bladder cancer, bone cancer, brain cancer, breast cancer, bronchial tumor, carcinoid tumor, heart tumor, cervical cancer, chordoma, chronic lymphocytic leukemia, chronic myeloproliferative neoplasm, colon cancer, colorectal cancer, craniopharyngioma, endometrial cancer, ependymoma, esophageal cancer, sensory neuroblastoma, Ewing sarcoma, eye cancer, fallopian tube cancer, gallbladder cancer, gastric carcinoid tumor, gastric stromal tumor, germ cell tumor, hairy cell leukemia, head and neck cancer, heart cancer, liver cancer, hypopharyngeal cancer, pancreatic cancer, kidney cancer, laryngeal cancer, chronic myeloid leukemia, lip and oral cancer, lung cancer, melanoma, Merkel cell carcinoma, mesothelioma, stomatal cancer, oral cancer, A pharmaceutical composition selected from osteosarcoma, ovarian cancer, penile cancer, pharyngeal cancer, prostate cancer, rectal cancer, salivary gland cancer, skin cancer, small intestine cancer, soft tissue sarcoma, testicular cancer, throat cancer, thyroid cancer, urethral cancer, uterine cancer, vaginal cancer, and vulvar cancer. Claim 6 A pharmaceutical composition according to claim 4, wherein the cancer is selected from breast cancer, colon cancer, rectal cancer, colorectal cancer, pancreatic cancer, and prostate cancer. Claim 7 A pharmaceutical composition according to claim 4, wherein the cancer is selected from hormone receptor-positive breast cancer, microsatellite-stable colon or rectal cancer, pancreatic cancer, and prostate cancer. Claim 8 delete Claim 9 delete Claim 10 delete Claim 11 delete Claim 12 delete Claim 13 delete Claim 14 delete Claim 15 delete Claim 16 delete Claim 17 delete

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