EA037780B1 - Nlrp3 modulators - Google Patents

Abstract

The invention relates to compounds of formula (II)or pharmaceutically acceptable salt thereof, wherein all the variables are as defined herein. These compounds modulate (e.g., agonize or partially agonize) NLRP3 that are useful, e.g., for treating a condition, disease or disorder in which an increase in an activity of NLRP3 may correct a deficiency in innate immune activity that contributes to the pathology and/or symptoms and/or progression and/or treatment refractory state of the condition, disease or disorder (e.g., cancers with low T-cell infiltration) in a subject (e.g., a human). The invention also relates to pharmaceutical compositions comprising the claimed compounds and the uses of the claimed compounds and the uses of the pharmaceutical compositions as a medicament and in treating cancer in a subject.

Description

Cross-reference to related claims

This application claims priority under US Provisional Patent Application No.

No. 62/460677, filed Feb. 17, 2017, U.S. Provisional Application No. 62/490881, filed April 2017, and U.S. Provisional Application No. 62/573991, filed Oct. 18, 2017, the contents of which are incorporated herein by reference in their entirety. completeness.

Field of the invention

The description refers to chemical entities (for example, a compound or a pharmaceutically acceptable salt and / or hydrate and / or co-crystal and / or drug combination of a compound) that modulate (for example, are agonists or partial agonists) NLRP3, which are useful, for example, for the treatment of a condition, disease or disorder in which an increase in NLRP3 activity can correct a deficiency in innate immune activity that contributes to the pathology and / or symptoms and / or progression and / or refractory to treatment of the condition, disease or disorder (for example, cancers with low infiltration of T cells) in a subject (eg, a human). The present disclosure also relates to compositions as well as other methods of using and preparing them.

State of the art

Nucleotide-binding oligomerization domain-like receptors (NLRs) include a family of intracellular receptors that exhibit pathogen-associated molecular patterns (PAMP) and endogenous molecules (cf. e.g. Ting, JPY et al., The NLR gene family: a standard nomenclature, Immunity, 28 (3): 285-287, (2008)).

NLRPs represent a subfamily of NLR receptors that include the Pyrin domain and include proteins such as NLRP1, NLRP3, NLRP4, NLRP6, NLRP7, and NLRP12. NLRPs are believed to be involved in the formation of multi-protein complexes called inflammasomes (see, for example, Chaput C. et al., NOD-like receptors in lung diseases, Frontiers in Immunology, 4: article 393, (2013)). These complexes usually include one or two NLR proteins, an apoptosis associated speck-like protein containing a CARD domain (ASC), an adapter speck-like protein associated with apoptosis, and procaspase-1F (see, for example, Bauernfeind F and Hornung V. Of inflammasomes and pathogens-sensing of microbes by the inflammasome, EMBO Molecular Medicine, 5 (6): 814-826, (2013)).

One of these inflammasomes is formed by the NLRP3 scaffold protein, the ASC adapter molecule, and procaspase-1 (see, for example, 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 Immunonogy, 129 (4): 1116.e6-1125.e6, (2012)), and its expression is believed to be induced by inflammatory cytokines and TLR agonists in myeloid cells and in humans in bronchial epithelial cells (Id. ). NLRP3 inflammasome is believed to mediate caspase-1-dependent conversion of pro-IL-1β and pro-III-18 to IL-1 and IL-18. In addition, IL-1e and IL-18 have potential in the treatment of various types of malignant diseases (see, for example, Chen L.-C. et al., EMBO Mol. Med., 4 (12): 1276-1293 (2012 ) and Tse BW-C. 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 (see, e.g., Ma Z. et al., Clin. Cancer Res. Jan 11, (2016) DOI: 10.1158 / 1078 -0432.CCR-15-1655).

The essence of the invention

The disclosure relates to chemicals (e.g., a compound or pharmaceutically acceptable salt and / or hydrate and / or co-crystal and / or drug combination of a compound) that modulate (e.g., are agonists or partial agonists) NLRP3, which are useful, for example, for the treatment of a condition, disease or disorder in which an increase in NLRP3 activity can correct the lack of innate immune activity that contributes to the pathology and / or symptoms and / or progression and / or refractory to treatment of the condition, disease or disorder (for example, malignant tumors with low infiltration of T cells) in a subject (eg, a human). The present disclosure also relates to compositions as well as other methods of using and preparing them.

The term NLRP3 agonist includes compounds that, at the protein level, directly bind or modify NLRP3 in such a way that the activity of NLRP3 is increased, for example, by activation, stabilization, altered distribution, or otherwise.

Certain compounds described herein that are less agonists of NLRP3 than a full NLRP3 agonist may function as antagonists as well as agonists in assays. These compounds antagonize the activation of NLRP3 by the full NLRP3 agonist as they prevent the full effect of the NLRP3 interaction. However, the compounds themselves also induce some NLRP3 activity, generally less than the corresponding amount of the full NLRP3 agonist. Such compounds can be referred to as partial agonists of NLRP3.

In some embodiments, the compounds described herein are

- 1,037780 agonists (eg full agonists) of NLRP3. In other embodiments, the compounds described herein are partial agonists of NLRP3.

Typically, the receptor exists in active (Ra) and inactive (Ri) conformation. Certain compounds that affect the receptor can alter the ratio of Ra to Ri (Ra / Ri). For example, a full agonist increases the Ra / Ri ratio and can produce a maximal, satiating effect. A partial agonist, when bound to a receptor, produces a response that is lower than that of a full agonist (eg, an endogenous agonist). Thus, the Ra / Ri for a partial agonist is less than for a full agonist. However, the potency of a partial agonist may be greater or less effective than that of a full agonist.

In one aspect, there are provided compounds of Formula II, or a pharmaceutically acceptable salt thereof

R ⁴ in which R ¹ , R ² , R ³ , R ⁴ , and R ^{5 are} defined below.

In one aspect, methods of modulating (e.g., agonizing, partially agonizing, antagonizing) NLRP3 activities are provided that comprise contacting NLRP3 with a chemical described herein (e.g., a compound described generally or specifically herein, or a pharmaceutically acceptable salt thereof, or compositions containing the same). In preferred embodiments, the methods for modulating NLRP3 activity are agonist and partially agonist. In certain embodiments, the methods for modulating NLRP3 activity are agonistic. In certain embodiments, the methods for modulating NLRP3 activity are partially agonistic. The methods include in vitro methods, for example, contacting a sample that includes one or more cells containing NLRP3 (eg, THP-1 cells) with a chemical. The methods can also include in vivo methods; for example, administration of a chemical to a subject (e.g., human) suffering from a disease in which an increase in NLRP3 activity can correct a deficiency in innate immune activity that contributes to the pathology and / or symptoms and / or progression of the disease (e.g., cancer; e.g., refractory cancer ).

In some embodiments, the compounds of the invention are useful for treating a condition, disease, or disorder in which a decrease in NLRP3 activity (e.g., a condition, disease, or disorder associated with suppressed or impaired NLRP3 signaling) contributes to the pathology and / or symptoms, and / or the progression of a condition, disease, or disorder (eg, cancer) in a subject (eg, human).

A malignant tumor is called refractory when it is not responsive to (or resistant to) treatment for the malignant tumor. Refractory cancer is also known as refractory cancer.

In another aspect, methods of treating cancer are provided which comprise administering to a subject in need of such treatment an effective amount of a chemical described herein (e.g., a compound described generally or specifically herein, or a pharmaceutically acceptable salt thereof, or compositions containing the same). In some embodiments, the cancer may be a refractory cancer.

In yet another aspect, there are provided methods of treating a disease in which an increase in NLRP3 activity can correct a deficiency in innate immune activity that contributes to the pathology and / or symptoms and / or progression of the disease, which include administering to a subject in need of such treatment an effective amount of a chemical substances described herein (for example, a compound described generally or specifically herein, or a pharmaceutically acceptable salt thereof, or compositions containing the same).

In another aspect, methods of treating a disease are provided that include administering to a subject having a disease in which an increase in NLRP3 activity can correct a deficiency in innate immune activity that contributes to the pathology and / or symptoms and / or progression of the disease, an effective amount of the chemical described herein (for example, a compound described generally or specifically herein, or a pharmaceutically acceptable salt thereof, or compositions containing the same).

In yet another aspect, methods of treating a disease are provided that include administering

- 2,037780 to the subject of a chemical described herein (for example, a compound described generally or specifically herein, or a pharmaceutically acceptable salt thereof, or compositions containing the same), wherein the chemical is administered in an amount effective to treat the disease in which an increase in NLRP3 activity can correct a deficiency in innate immune activity that contributes to the pathology and / or symptoms and / or progression of the disease, thereby providing treatment for the disease.

Embodiments may include one or more of the following features.

The chemical can be administered in combination with one or more additional anti-cancer therapies (for example, surgery, radiation therapy, chemotherapy, toxotherapy, immunotherapy, cryotherapy, or gene therapy, or a combination thereof; for example, anti-cancer therapies that include the administration of one or more (for example, two, three, four, five, six or more) additional anti-cancer agents Non-limiting examples of additional anti-cancer agents (chemotherapeutic agents) are selected from an alkylating agent (e.g., cisplatin, carboplatin, mechlorethamine, cyclophosphamide, chlorambucil, ifosfamide and / or oxaliplatin), an antimetabolite (e.g. azathioprine and / or mercaptopurine), terpenoid (e.g. vinca alkaloid and / or taxane, e.g. vincristine, vinblastine, vinorelbine and / or vindesine taxol, paclitaxel and / or docetaxel), topoisomerase (e.g., type I topoisomerase and topoisomerase type 2; e.g. camptothecins such as irinotecan and / or topotecan, amsacrine, etoposide, etoposite phosphate and / or teniposide); a cytotoxic antibiotic (eg, actinomycin, anthracyclines, doxorubicin, daunorubicin, valrubicin, idarubicin, epirubicin, bleomycin, plikamycin, and / or mitomycin); a hormone (eg, a luteinizing hormone releasing hormone agonist; eg, leuprolidine, goserelin, triptorelin, histrelin, bicalutamide, flutamide and / or nilutamide); antibodies (e.g., Abciximab, Adalimumab, Alemtuzumab, Atlizumab, Atlizumab) Basiliximab, Belimumab, Bevacizumab (Brevacentizabotumab) , Cetuximab, Ceertolizumab pegol (Ceertolizumab pegol) (Daclizumab), Denosumab, Eculizumab, Golizumab, Golizumab, Golizumabolab, Gemtuzumab (Ibritumomab tiuxetan), Infliximab (Infliximab), Ipilimumab (Ipilimumab), Muromonab-CD3 (Muromonab-CD3), Natalizumab, Ofatumumab (Ofatumumab), Omalizumab (Omalizumabitumab) (Omalizumabumab) (Ranibizumab), Rituximab, Tocilizumab, Tositumomab and / or Trastuzumab); anti-angiogenic agent; cytokine; a thrombotic agent; a growth inhibiting agent; anthelmintic agent; and an immune checkpoint inhibitor that targets an immune checkpoint receptor selected from 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 (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, B7-H3, B7-H4, VISTA, TMIGD2, HHLA2TMIGD2, butyrophilins including BTNL2, Sigitlec family, members of the TIGITlec family and PVR, 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, neuropilin, CD160 , CD30 and Cd155 (e.g. CTLA-4 or PD1 or PD-L1) and other immunomodulatory agents such as interleukin-2 (IL-2), indoleamine-2,3-dioxygenase (IDO), IL-10, transforming growth factor-β (TGFe), CD39, CD73 Adenosine-CD39-CD73 and CXCR4-CXCL12 ...

The subject may have cancer; for example, a subject has undergone, and / or is under, and / or will undergo one or more methods of treating a malignant disease.

Non-limiting examples of malignant disease include acute myeloid leukemia, adrenocortical cancer, Kaposi's sarcoma, lymphoma, anal cancer, appendix cancer, teratoid / rhabdoid tumor, basal cell carcinoma, bile duct cancer, bladder cancer, bone cancer, brain cancer, breast cancer, bronchial tumor, carcinoid tumor, cardiac tumor, cervical cancer, chordoma, chronic lymphocytic leukemia, chronic myeloproliferative neoplasms, colon cancer, colorectal cancer, craniopharyngioma, bile duct cancer, endometrial cancer, ependymoma, esophageal cancer, esthesioneurobarkoma ), eye cancer, fallopian tube cancer, gallbladder cancer, gastrointestinal carcinoid tumors, gastrointestinal stromal tumors, germ cell tumors, hair cell leukemia, head and neck cancer, heart cancer, liver cancer, hypopharyngeal cancer, pancreatic cancer , kidney cancer, laryngeal cancer, chronic myelogenous leukemia, lip cancer, etc. Oral cavity, lung cancer, melanoma, Merkel carcinoma, mesothelioma, oral cancer, oral cancer, osteosarcoma, ovarian cancer, penile cancer, pharyngeal cancer, prostate cancer, rectal cancer, salivary gland cancer, skin cancer, small bowel cancer , soft tissue sarcoma, testicular cancer, throat cancer, thyroid cancer, urethral cancer, uterine cancer, cancer

- 3 037780 vagina and vulvar cancer.

In other embodiments, the mammal has been identified as having cancer or an infectious disease. Representative infectious diseases include, but are not limited to, Acinobacter infection, actinomycosis, African sleeping sickness, acquired immunodeficiency syndrome, amebiasis, anaplasmosis, anthrax, Arcanobacterium haemonyticum infection, Argentine hemorrhagic fever, Bacteriosis, Bacteriosis, aspergillosis pneumonia, bacterial vaginosis, Bacteroides infection, balantidiasis, Baylisascaris infection, BK viral infection, black piedra, Blastocystic hominis infection, blastomycosis, Bolivian hemorrhagic fever, botulism, Brazilian haemorrhagic fever, Burholis, brucellosis Calicivirus infection, camptobacteriosis, candidiasis, cat scratch disease, cellulitis, Chagas disease, chancroid, chickenpox, chikungunya, chlamydia, Chlamydophila pneumoniae infection, cholera, chromoblastomycosis, clonorchiasis, Clostridium difficile infection, clostridium difficile Olorada tick-borne fever, viral infection of the upper respiratory tract, Creutzfeldt-Jakob disease, Crimean-Congo hemorrhagic fever, crytococcosis, cryptosporidiosis, migratory skin larva, Cyclosporosis, cysticercosis cysticercosis , diphyllobothriasis, dracunculiasis, hemorrhagic fever Ebola, echinococcosis, ehrlichiosis, enterobiasis, Enterococcus infection, Enterovirus infection, epidemic typhus, erythema infection, exanthema, fascioloptiosis, fasciolosis, fatal familial insomnia amoebae, Fusobacterium infection, gas gangrene, geotrichosis, Gerstmann-Straussler-Scheinker syndrome, giardiosis, glanders, gnatostomosis, gonorrhea, inguinal granuloma, streptococcal infection of group A, streptococcal influenza ilious vesicular stomatitis, hantavirus pulmonary syndrome, Heartland virus disease, Heliobacter pylori infection, hemolytic uremic syndrome, hemorrhagic fever with renal syndrome, hepatitis A, hepatitis B, hepatitis C, hepatitis D, hepatitis E, herpes simplex, histoplasmosis , human bocavirus infection, human ehrlichiosis, anaplasmosis of human granulocytes, human metaplevomyovirus infection, human monocytic ehrlichiosis, human papillomavirus infection, human parainfluenza virus infection, hymenolepasis, Epstein-Barr virus (Epstein-Barr disease), infectious mononucleus , keratitis, Kingella kingae infection, kuru, Lassa fever, Legionnaires' disease, Pontiac fever, leishmaniasis, leprosy, leptospirosis, listeriosis, Lyme disease, lymphatic filariasis, lymphocytic choreomeningitis, malaria, Marburg hemorrhagic fever, mild pea syndrome , meningoc ocular infection, metagonimiasis, microsporidiosis, molluscum contagiosum, monkeypox, mumps, rat typhus, mycoplasma pneumonia, mycetoma, myiasis, neonatal conjunctivitis, a variant of the Creutzfeldt-Jakob disease, nocardiosis, parasymocymidosis, headache pediculosis, pubic lice, pelvic inflammatory disease, pertussis, plague, pneumonia, poliomyelitis, Prevotella infection, primary amoebic meningoencephalopathy, progressive multifocal leukoencephalopathy, psittacosis, Q fever, rabies, relapsing rheumatic fever infection, rickettsiosis, Rift Valley fever, Rocky Mountain spotted fever, rotavirus infection, rubella, salmonellosis, severe acute respiratory syndrome, scabies, schistosomiasis, sepsis, shigellosis, herpes zoster, smallpox, sporotrichocosis, staphylococcus aureus tion, staphylococcal infection, strongyloidosis, subacute sclerosing panencephalitis, syphilis, teniasis, tetanus, ringworm barabe, ringworm, fungal infection of the smooth skin of the trunk, inguinal dermatomycosis, fungal infection of the skin of the hands, black lichen, fungal fungal infection versicolor, toxocariasis, trachoma, toxoplasmosis, trichinosis, trichomoniasis, trichuriasis, tuberculosis, tularemia, typhoid fever, Ureaplasma urealyticum infection, valley fever, Venezuelan hemorrhagic fever, viral pneumonia, pyretic fever, western pus , yellow fever and zygomycosis.

The chemical can be administered intratumorally.

The chemical can be administered systemically (including, but not limited to, oral, subcutaneous, intramuscular, intravenous).

The methods can further include identifying the subject.

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

Additional definitions.

To facilitate an understanding of the disclosure set forth herein, a number of additional terms are defined below. Typically, the nomenclature used in this document and the laboratory procedures in organic chemistry, medicinal chemistry and pharmacology described in this

- 4037780 document, are well known and widely used in the art. Unless otherwise indicated, all technical and scientific terms used in this document generally have the same meaning as commonly understood by those of ordinary skill in the art to which this disclosure relates.

As used herein, the term NLRP3 includes, without limitation, nucleic acids, polynucleotides, oligonucleotides, sense and antisense strands of polynucleotides, complementary sequences, peptides, polypeptides, proteins, homologous and / or orthologous NLRP3 molecules, isoforms, precursors, mutants, variants , derivatives, splicing variants, alleles, various species and their active fragments.

As used herein, the term “acceptable” in relation to a formulation, composition, or ingredient means no permanent deleterious effect on the general health of the subject being treated.

API refers to an active pharmaceutical ingredient.

The terms effective amount or therapeutically effective amount, as used herein, refers to a sufficient amount of a chemical (e.g., a compound that exhibits activity as a mitochondria-uncoupling agent, or a pharmaceutically acceptable salt and / or hydrate and / or co-crystal thereof, for example a compound, such as niclosamide or a pharmaceutically acceptable salt and / or hydrate and / or co-crystal thereof, for example a compound such as a niclosamide analog or a pharmaceutically acceptable salt and / or hydrate and / or co-crystal), which is administered to some extent will ease one or more symptoms of the disease or condition being treated. The result includes a reduction and / or mitigation of signs, symptoms or causes of disease or any other desired change in the biological system. For example, an effective amount for therapeutic use is the amount of a composition containing a compound as described herein required to provide a clinically significant reduction in disease symptoms. The appropriate effective amount in each case is determined using any suitable method, such as a dose escalation study.

The term excipient or pharmaceutically acceptable excipient means a pharmaceutically acceptable material, composition or carrier, for example, a liquid or solid filler, diluent, carrier, solvent or encapsulating material. In one embodiment, each component is pharmaceutically acceptable in the sense of being compatible with the other ingredients of the pharmaceutical composition and suitable for use in contact with a tissue or organ of humans and animals without undue toxicity, irritation, allergic response, immunogenicity, or other problems or complications commensurate with a reasonable ratio benefit / risk. See, for example, 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 .; Gibson Ed .; CRC Press LLC: Boca Raton, FL (2009).

The term pharmaceutically acceptable salt refers to a formulation of a compound that does not cause significant irritation to the host to which it is administered, and does not eliminate the biological activity and properties of the compound. In some cases, pharmaceutically acceptable salts are prepared by reacting a compound described herein with acids such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid, salicylic acid, and the like. In some cases, pharmaceutically acceptable salts are prepared by reacting a compound having an acidic group as 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 salt or magnesium, a salt of organic bases such as dicyclohexylamine, N-methyl-D-glucamine, tris (hydroxymethyl) methylamine, and salts with amino acids such as arginine, lysine and the like, or by other methods as previously defined. The pharmaceutically acceptable salt is not particularly limited as far as it can be used in medicaments. Examples of the salt that the compounds described herein form with a base include the following: their salts with inorganic bases such as sodium, potassium, magnesium, calcium, and aluminum; their salts with organic bases such as methylamine, ethylamine and ethanolamine; their salts with basic amino acids such as lysine and ornithine; and ammonium salt. Salts can be acid addition salts, examples of which are acid addition salts with the following: mineral acids such as hydrochloric acid, hydrobromic acid, hydroiodic acid, 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; acidic amino acids 5,037,780 such as aspartic acid and glutamic acid.

The term pharmaceutical composition refers to a mixture of a compound described herein with other chemical components (collectively referred to herein as adjuvants) such as carriers, stabilizers, diluents, dispersing agents, suspending agents and / or thickening agents. The pharmaceutical composition facilitates administration of the compound to the body. Several routes of administration of the compound exist in the art, including, but not limited to: rectal, oral, intravenous, aerosol, parenteral, ocular, pulmonary, and topical administration.

The term “subject” refers to an animal, including, but not limited to, a primate (eg, human), monkey, cow, pig, sheep, goat, horse, dog, cat, rabbit, rat, or mouse. The terms subject and patient are used interchangeably herein to refer to, for example, a mammalian subject such as a human.

The terms treat and treat in the context of treating a disease or disorder are intended to include amelioration or elimination of a disorder, disease or condition, or one or more symptoms associated with the disorder, disease or condition; or slowing the progression, spread, or worsening of a disease, disorder, or condition, or one or more of its symptoms. Cancer treatment refers to one or more of the following effects: (1) inhibiting, to some extent, tumor growth, including (i) slowing down and (ii) stopping growth completely; (2) a decrease in the number of tumor cells; (3) maintaining tumor size; (4) reducing the size of the tumor; (5) inhibiting, including (i) reducing, (ii) slowing down or (iii) completely preventing the infiltration of tumor cells into peripheral organs; (6) inhibition, including (i) reduction, (ii) delay, or (iii) complete prevention of metastasis; (7) enhancement of the anti-tumor immune response, which can lead to (i) maintenance of tumor size, (ii) reduction of tumor size, (iii) slowing tumor growth, (iv) shrinking, slowing down or preventing invasion, and / or (8) relief, to some extent, the severity, or the amount of one or more symptoms associated with the disorder.

The term halogen refers to a fluorine (F), chlorine (Cl), bromine (Br) or iodine (I) atom.

The term alkyl refers to a hydrocarbon chain, which may be straight or branched, containing the indicated number of carbon atoms. For example, C _1-10 indicates that the group may contain from 1 to 10 (inclusive) carbon atoms therein. Non-limiting examples include methyl, ethyl, isopropyl, t-butyl, n-hexyl.

The term haloalkyl refers to n-alkyl in which one or more hydrogen atoms are / are substituted by an independently selected halogen atom.

The term alkoxy refers to an —O-alkyl radical (eg —OCH ₃ ).

The term alkylene refers to a branched or unbranched divalent alkyl (eg, -CH2-).

The term alkenyl refers to a hydrocarbon chain that can be straight or branched chain having one or more carbon-carbon double bonds. The alkenyl part contains the indicated number of carbon atoms. So, for example, C _2-6 indicates that the group may have from 2 to 6 carbon atoms (inclusive) in it.

The term alkynyl refers to a hydrocarbon chain that can be straight or branched chain having one or more carbon-carbon triple bonds. The alkynyl moiety contains the indicated number of carbon atoms. So, for example, C2-6 indicates that the group may have from 2 to 6 carbon atoms (inclusive) in it.

The term aryl refers to a 6-carbon monocyclic, 10-carbon bicyclic or 14-carbon tricyclic aromatic ring system in which 0, 1, 2, 3, or 4 atoms of each ring may be substituted with a substituent, and where the ring containing the monocyclic radical is aromatic, and wherein at least one of the condensed rings containing a bicyclic or tricyclic radical is aromatic, for example tetrahydronaphthyl. Examples of aryl groups also include phenyl, naphthyl, and the like.

As used herein, the term cycloalkyl includes saturated cyclic hydrocarbon groups having 3 to 10 carbon atoms, preferably 3 to 8 carbon atoms, and more preferably 3 to 6 carbon atoms, said cycloalkyl group being optionally substituted. Preferred cycloalkyl groups include, without limitation, cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl, cycloheptyl, and cyclooctyl. As used herein, the term cycloalkylene refers to a divalent cycloalkyl.

The term heteroaryl refers to an aromatic 5 to 8 membered monocyclic, 8 to 12 membered bicyclic, or 11 to 14 membered tricyclic ring system having 1 to 3 heteroatoms when monocyclic, 1 to 6 heteroatoms when bicyclic, or 1 to 9 heteroatoms in the case of a tricyclic system, said heteroatoms being selected from O, N or S (e.g. carbon atoms and 1 to 3, 1 to 6, or 1 to 9 heteroatoms N, O or S in the case of mono

- 6 037780 cyclic, bicyclic or tricyclic system, respectively), where 0, 1, 2, 3 or 4 atoms of each ring may be substituted with a substituent, and where the ring containing the monocyclic radical is aromatic, and where at least one of the condensed rings containing a bicyclic or tricyclic radical is aromatic (but need not be a ring that contains a heteroatom, for example, tetrahydroisoquinolinyl. Examples of heteroaryl groups also include pyridyl, furyl or furanyl, imidazolyl, benzimidazolyl, pyrimidinyl, thiophenyl or thienyl, indolinyl, thiophenyl etc.

The term heterocyclyl refers to a non-aromatic 5 to 8 membered monocyclic, 8 to 12 membered bicyclic, or 11 to 14 membered tricyclic ring system having 1 to 3 heteroatoms when monocyclic, 1 to 6 heteroatoms when bicyclic, or from 1 to 9 heteroatoms in the case of a tricyclic system, said heteroatoms being selected from O, N or S (for example, carbon atoms and from 1 to 3, from 1 to 6 or from 1 to 9 heteroatoms N, O or S in the case of a monocyclic , bicyclic or tricyclic system, respectively), where 0, 1, 2 or 3 atoms of each ring may be substituted with a substituent. Examples of heterocyclyl groups include piperazinyl, pyrrolidinyl, dioxanyl, morpholinyl, tetrahydrofuranyl, and the like. The term heterocycloalkylene refers to a divalent heterocyclyl.

In addition, the atoms constituting the compounds of these embodiments are intended to include all isotopic forms of such atoms. Isotopes as used herein include those atoms that have 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 include ¹³ C and ¹⁴ C.

Details of one or more embodiments of the invention are set forth in the accompanying drawings and in the description below. Other features and advantages of the invention will be apparent from the description and drawings, as well as from the claims.

Detailed description

The invention provides chemicals (e.g., a compound or pharmaceutically acceptable salt and / or hydrate and / or co-crystal and / or drug combination of a compound) that modulate (e.g. are agonists or partial agonists) NLRP3 that are useful, e.g. for the treatment of a condition, disease or disorder in which an increase in NLRP3 activity can correct a lack of innate immune activity (for example, a condition, disease or disorder associated with an insufficient immune response) that contributes to the pathology and / or symptoms and / or progression of the condition, a disease or disorder (eg, cancer) in a subject (eg, human). This disclosure also contains compositions, as well as other methods of their use and preparation.

Compounds of formula II.

The invention relates to a compound of formula (II)

or a pharmaceutically acceptable salt thereof, where

R ^{1 is} independently unsubstituted C _1-6 αalkyl, C (= O) R ^a ;

R ^{2 is} independently H or unsubstituted C _1-6 α-alkyl;

R ³ represents:

(i) H;

(ii) unsubstituted C _1-2 alkyl;

(iii) XR ⁸ wherein X is unbranched C1-6 alkylene and R ⁸ is —OH, C1-4 alkoxy, —C _1-4 haloalkoxy, CO ₂ R ^a or —CONR ^c R ^d ;

(iv) (C _1-3 alkylene) (C ₆ -C ₁₀ aryl) wherein said aryl is optionally substituted with 1 to 3 substituents independently selected from C _1-6 alkyl, C _1-4 haloalkyl, C _1-4 alkoxy and C _1-4 haloalkoxy; or (v) (C _1-3 alkylene) heteroaryl having 5 to 6 ring atoms, where each of 1 to 4 ring atoms is independently selected from the group consisting of N, N (R ^e ), O and S, and wherein said heteroaryl is optionally substituted with 1 to 3 substituents independently selected from the group consisting of: C _1-6 alkyl, C _1-4 haloalkyl, C _1-4 α alkoxy, and C _1-4 haloalkoxy;

R ⁴ and R ⁵ are each independently selected from:

(i) H;

- 7 037780 (ii) halogen;

(iii) - (C _0-3 alkylene) C _3-10 cycloalkyl, said cycloalkyl optionally substituted with 1 to independently selected Rf;

(iv) - (C _0-3 alkylene) heterocyclyl containing from 3 to 10 ring atoms, and from 1 to 3 ring atoms, each independently selected from the group consisting of N (R ^e ), O and S, where said heterocyclyl is optionally substituted with 1 to 4 independently selected R ^f ;

(v) - (C _0-3 alkylene) (C ₆ -C ₁₀ aryl) optionally substituted with 1 to 4 R ^g ;

(vi) - (C _0-3 alkylene) heteroaryl having 5 to 10 ring atoms, with 1 to 4 ring atoms each independently selected from the group consisting of N, NHO and S, wherein said heteroaryl is optionally substituted from 1 to 3 R ^g ;

(vii) Cl _-6 alkyl optionally substituted with 1 to 2 independently selected R ^h ; and (viii) - (C _0-3 alkylene) C _{4-1 0} cycloalkenyl, said cycloalkenyl optionally substituted with 1 to 2 R ^f ;

R ^a represents:

(i) C _1-6 alkyl optionally substituted with 1 to 2 R ^h ;

(ii) - (C _0-3 alkylene) C _3-10 cycloalkyl, said cycloalkyl optionally substituted with 1 to 2 R ^f ;

(iii) - (C _1-3 alkylene) heterocyclyl having 3 to 10 ring atoms, wherein 1 to 3 ring atoms are each independently selected from N (R ^e ), O and S, wherein said heterocyclyl is optionally substituted 1 to 4 independently selected R ^f ;

(iv) - (C _0-3 alkylene) phenyl optionally substituted with 1 to 4 independently selected R ^g ; or (v) - (C _0-3 alkylene) heteroaryl having 5 to 10 ring atoms, with 1 to 4 ring atoms each independently selected from the group consisting of N, N (R ^e ), O and S, where said heteroaryl is optionally substituted with R ^g ;

in each case, R ^c and R ^{d are} independently H or C _1-4 alkyl;

in each case, R ^{e is} independently H or C _1-4 alkyl;

in each occurrence, Rf is independently C _1-6 alkyl, C _1-4 haloalkyl, —OH, F, Cl, C _1-4 alkoxy, C _1-4 haloalkoxy, cyano, or phenyl optionally substituted with 1 to 4 R ^g ;

in each occurrence, R ^{g is} independently halogen, cyano, C _1-6 alkyl, C _1-4 haloalkyl, C _1-4 alkoxy, or C _1-4 haloalkoxy; and in each occurrence, R ^{h is} independently —OH, F, C _1-4 alkoxy, C _1-4 haloalkoxy, or cyano.

In another aspect, the present invention relates to a compound of formula (II), or a pharmaceutically acceptable salt thereof, wherein

R ³ is H, unsubstituted C1-2 alkyl or XR ⁸ , wherein X is unbranched C1-2 alkylene and R ⁸ is CO ₂ R ^a or —CONR ^c R ^d ;

R ⁴ independently represents H or halogen;

R ^{5 is} independently selected from:

(i) H;

(ii) halogen;

(iii) - (C _0-3 alkylene) C _3-10 cycloalkyl, said cycloalkyl optionally substituted with 1 to 4 independently selected R ^f ;

(iv) - (C _0-3 alkylene) heterocyclyl containing from 3 to 10 ring atoms, and from 1 to 3 ring atoms, each independently selected from the group consisting of N (R ^e ), O and S, where said heterocyclyl is optionally substituted with 1 to 4 R ^f ;

(v) - (C _0-3 alkylene) (C ₆ -C ₁₀ aryl) optionally substituted with 1 to 4 R ^g ;

(v) - (C _0-3 alkylene) heteroaryl having 5 to 10 ring atoms, with 1 to 4 ring atoms each independently selected from the group consisting of N, NH, O and S, wherein said heteroaryl is optionally substituted with 1 to 3 R ^g ;

(vi) C _1-6 alkyl optionally substituted with 1 to 2 independently selected R ^h ; and (vii) - (C _0-3 alkylene) C _4-10 cycloalkenyl, said cycloalkenyl optionally substituted with 1 to 2 R ^f ;

In another aspect, the present invention relates to a compound of formula (II), or a pharmaceutically acceptable salt thereof, wherein

R ^{1 is} independently unsubstituted C _1-6 alkyl, C (= O) R ^a ;

R ² independently represents H or unsubstituted C1-3alkyl;

R ³ is H, unsubstituted C _1-2 alkyl or XR ⁸ , wherein X is unbranched C 2-4 alkylene and R ⁸ is CO2R ^a or —CONR ^c R ^d ;

R ^{5 is} independently selected from:

(i) C3-6cycloalkyl optionally substituted with 1 to 2 independently selected R ^f ;

(ii) phenyl optionally substituted with 1 to 3 R ^g ;

(iii) heteroaryl containing from 5 to 6 ring atoms, with from 1 to 3 ring atoms,

- 8 037780 each independently selected from N, NH, O and S, wherein said heteroaryl is optionally substituted from 1 to

R ^g ;

(iv) C _1-6 alkyl optionally substituted with 1 to 2 independently selected R ^h ; and (v) C _5-6 cycloalkenyl optionally substituted with 1 to 2 Rf;

R ^a is Cl _-4 alkyl optionally substituted with OH, C _3-6 cycloalkyl, phenyl or heteroaryl having 5 to 6 ring atoms, where 1 to 4 ring atoms are each independently selected from N, N (R ^e ), O and S.

In another aspect, the present invention relates to a compound of formula (II), or a pharmaceutically acceptable salt thereof, wherein

R ^{1 is} independently Cl _-6 alkyl, C (= O) R ^a ;

R ^{2 is} independently H, CH ₃ or CH ₂ CH ₃ ;

R ³ represents H, CH ₃ or - (CH ₂ ) ₃ C (= O) OCH ₃ ;

R ^{5 is} independently CH3, cyclopentyl, cyclopentenyl, phenyl, pyrazol-1-yl, or pyrazol-3-yl; and

R ^a is CH ₃ , CH2CH3, CH (CH ₃ ) ₂ , C (CH ₃ ) ₃ , cyclopropyl or thiazolyl.

In another aspect, the present invention relates to a compound of formula (II), or a pharmaceutically acceptable salt thereof, wherein

R ^{1 is} independently Cl _-6 alkyl, C (= O) R ^a ;

R ^{2 is} independently H, CH ₃ or CH ₂ CH ₃ ;

R ³ represents H or CH ₃ ;

R ^{5 is} independently CH3, cyclopentyl, cyclopentenyl, phenyl, pyrazol-1-yl, or pyrazol-3-yl; and

R ^a represents CH3, CH2CH3, CH (CH3) 2, C (CH3) 3 or cyclopropyl.

In another aspect, the present invention relates to a compound of formula (II), or a pharmaceutically acceptable salt thereof, wherein

R ^{1 is} independently C (= O) R ^a ;

R ^{2 is} independently H, CH ₃ or CH ₂ CH ₃ ;

R ³ represents H;

R ^{5 is} independently cyclopentyl, cyclopentenyl, phenyl, pyrazol-1-yl, or pyrazol3-yl; and

R ^a is CH ₃ , CH2CH3, CH (CH ₃ ) ₂ , C (CH ₃ ) _3, or cyclopropyl.

In another aspect, the present invention relates to a compound of formula (II), or a pharmaceutically acceptable salt thereof, wherein

R ¹ independently represents C (CH ₃ ) ₃ ;

R ² independently represents H, CH3, or CH2CH3;

R ³ independently represents H, CH ₃ or CH ₂ CH ₃ ;

R ⁴ represents H; and

R ⁵ independently represents pyrazol-1-yl, pyrazol-3-yl, or pyrazol-5-yl.

In another aspect, the present invention relates to a compound of formula (II), or a pharmaceutically acceptable salt thereof, wherein

R ¹ represents C (= O) R ^a ;

R ^{2 is} independently H, CH _3, or CH2CH3;

R ³ independently represents H, CH ₃ , CH2CH3, or CH2CH2OH;

R ⁴ represents H;

R ^{5 is} independently cyclopentyl, cyclopentenyl, thienyl, pyrazol-1-yl, pyrazol-3yl, pyrazol-5-yl, or (phenyl substituted with 0 to 1 C _1-4 alkyl); and

R ^a is CH ₃ , CH2CH3, CH (CH ₃ ) ₂ , C (CH ₃ ) ₃ , - (CH) ₂ CH (CH ₃ ) ₂ , cyclopropyl, 1-methyl1H-pyrrol-2-yl or (phenyl, substituted with C _1-4 alkoxy or Cl).

In some aspects, R ¹ is unsubstituted C _1-6 alkyl or C (= O) OR ^a .

In some aspects, R ² is H or unsubstituted C 1-6 alkyl. In other aspects, R ² is H or unsubstituted C1-3alkyl. In other aspects, R ² is H, CH3, or CH2CH3. In other aspects, R ² is H. In other aspects, R ² is CH3 or CH2CH3. In other aspects, R ² is CH ₃ . In other aspects, R ² is CH 2 CH 3.

In some aspects, R ³ is H, unsubstituted C 1-2 alkyl, or XR ⁸ , wherein X is unbranched C 2-6 alkylene and R ⁸ is C02R ^a or —CONR ^c R ^d . In other aspects, R ³ is H, unsubstituted C 1-2 alkyl, or XR ⁸ , wherein X is unbranched C 2-4 alkylene and R ⁸ is C02R ^a or —CONR ^c R ^d . In other aspects, R ³ is H, CH3, or - (CH2) 3C (= O) OCH3. In other aspects, R ³ is H or unsubstituted C1-2 alkyl. In other aspects, R ³ is H or CH3. In other aspects, R ³ is H. In other aspects, R ³ is CH3.

In some aspects, R ⁴ is H or halogen. In other aspects, R ⁴ is co-9 037780 H.

In some aspects, R ^{5 is} independently selected from: (i) - (C 0-3 alkylene) C 3-10 cycloalkyl, said cycloalkyl optionally substituted with 1 to 4 independently selected R ^f ; (ii) - (C0-3alkylene) heterocyclyl having 3 to 10 ring atoms, with 1 to 3 ring atoms each independently selected from: N (R ^e ), O and S, wherein said heterocyclyl is optionally substituted with 1 up to 4 R ^f ; (iii) - (C0-3 alkylene) (C6-C10 aryl) optionally substituted with 1 to 4 R ^g ; (iv) - (C0-3alkylene) heteroaryl having 5 to 10 ring atoms, with 1 to 4 ring atoms each independently selected from: N, NH, O and S, wherein said heteroaryl is optionally substituted with 1 to 3 R ^g ; (v) C1-6alkyl optionally substituted with 1 to 2 independently selected R ^h ; and (iv) - (C0-3alkylene) C ₃ 10 cycloalkenyl, said cycloalkenyl optionally substituted with 1 to 2 R ^f .

In other aspects, R ^{5 is} independently selected from: (i) C3-6 cycloalkyl optionally substituted with 1 to 2 independently selected R ^f ; (ii) phenyl optionally substituted with 1 to 3 R ^g ; (iii) heteroaryl having 5 to 6 ring atoms, with 1 to 3 ring atoms each independently selected from: N, NH, O and S, wherein said heteroaryl is optionally substituted with 1 to 3 R ^g ; (iv) C1-6alkyl optionally substituted with 1 to 2 independently selected R ^h ; and (v) C _5-6 cycloalkenyl optionally substituted with 1 to 2 R ^f .

In other aspects, R ^{5 is} independently CH ₃ , cyclopentyl, cyclopentenyl, phenyl, pyrazol-1-yl, or pyrazol-3-yl. In other aspects, R ⁵ is CH3. In other aspects, R ⁵ is cyclopentyl or cyclopentenyl. In other aspects, R ⁵ is cyclopentyl. In other aspects, R ⁵ is cyclopentenyl. In other aspects, R ⁵ is phenyl. In other aspects, R ⁵ is pyrazolyl. In other aspects, R ⁵ is pyrazol-1-yl or pyrazol-3-yl. In other aspects, R ⁵ is pyrazol-1-yl. In other aspects, R ⁵ is pyrazol-3-yl.

In some aspects (i) C _1-6 alkyl optionally substituted with 1 to 2 R ^h ; (ii) - (C0-3alkylene) C3-10cycloalkyl, said cycloalkyl optionally substituted with 1 to 2 R ^f ; (iii) - (C0-3alkylene) phenyl optionally substituted with 1 to 4 independently selected R ^g ; or (v) - (C0-3alkylene) heteroaryl having 5 to 10 ring atoms, wherein 1 to 4 ring atoms are each independently selected from N, N (R ^e ), O and S, the heteroaryl optionally substituted with 1 to 3 independently selected R ^g . In other aspects, R ^a is (i) C1-6 alkyl optionally substituted with 1 to 2 R ^h ; (ii) - (C0-3alkylene) C3-10cycloalkyl, said cycloalkyl optionally substituted with 1 to 2 R ^f . In other aspects, R ^a is H, C1-4 alkyl optionally substituted with OH, C _3-6 cycloalkyl, phenyl, or heteroaryl having 5 to 6 ring atoms, where 1 to 4 ring atoms are each independently selected from NN (R ^e ), O, and S. In other aspects, R ^a is C1-6 alkyl optionally substituted with 1 to 2 R ^h. In other aspects, R ^a is C1-6 alkyl. In other aspects, R ^a is - (C0-3alkylene) C3-10cycloalkyl, said cycloalkyl optionally substituted with 1 to 2 Rf. In other aspects, R ^a is —C3-10cycloalkyl, optionally substituted with 1 to 2 Rf. In other aspects, R ^a is —C3-6cycloalkyl. In other aspects, R ^a is C1-4 alkyl or C _3-6 cycloalkyl. In other aspects, R ^a is CH3, CH2CH3, CH (CH3) 2, or cyclopropyl. In other aspects, R ^a represents CH3, CH2CH3 or CH _{(CH3) 2.} In other aspects, R ^a is cyclopropyl.

In another aspect, a compound of the invention is selected from

- 10 037780

- 12 037780

or a pharmaceutically acceptable salt thereof.

In another aspect, a compound of the invention is selected from

- 13 037780

or a pharmaceutically acceptable salt thereof.

In another aspect, a compound of the invention is selected from

- 14 037780

- 15 037780

- 16 037780

or a pharmaceutically acceptable salt thereof.

In another aspect, a compound of the invention is selected from

- 17 037780 or a pharmaceutically acceptable salt thereof.

In another aspect, a compound of the invention is selected from

or a pharmaceutically acceptable salt thereof.

In another aspect, a compound of the invention is selected from

- 18 037780

or a pharmaceutically acceptable salt thereof.

In another aspect, a compound of the invention is selected from

- 19 037780

or a pharmaceutically acceptable salt thereof.

In another aspect, a compound of the invention is selected from

- 20 037780

or a pharmaceutically acceptable salt thereof.

In another aspect, a compound of the invention is selected from

- 21 037780

or a pharmaceutically acceptable salt thereof.

In another aspect, a compound of the invention is

or a pharmaceutically acceptable salt thereof.

In another aspect, a compound of the invention is

or a pharmaceutically acceptable salt thereof.

In another aspect, a compound of the invention is

or a pharmaceutically acceptable salt thereof.

In another aspect, a compound of the invention is

- 22 037780

or a pharmaceutically acceptable salt thereof.

In another aspect, a compound of the invention is

or a pharmaceutically acceptable salt thereof.

In another aspect, a compound of the invention is

or a pharmaceutically acceptable salt thereof.

In another aspect, the invention relates to a compound of formula (I), or a pharmaceutically acceptable salt thereof

with R ⁴ (I) where R ^{1 is} independently unsubstituted C _1-6 α-alkyl or C (= O) R ^a ;

R ^{2 is} independently H or unsubstituted C _1-6 α-alkyl;

R ³ represents:

ω ^H; (ii) unsubstituted C _1-2 alkyl;

(iii) XR ⁸ wherein X is unbranched C1-6αalkylene and R ⁸ is —OH, C1-4alkoxy, —C _1-4 haloalkoxy, CO ₂ R ^a, or —CONR ^c R ^d ;

(iv) (C _1-3 alkylene) (C ₆ -C ₁₀ aryl), wherein aryl is optionally substituted with 1 to 3 substituents independently selected from C _1-6 alkyl, C _1-4 haloalkyl, C _1-4 alkoxy and C _1-4 haloalkoxy; or (v) (C _1-3 alkylene) heteroaryl having 5 to 6 ring atoms, where 1 to 4 ring atoms are each independently selected from N, N (R ^e ), O and S, and where heteroaryl is optionally substituted from 1 to 3 substituents independently selected from C _1-6 alkyl, C _1-4 haloalkyl, C _1-4 alkoxy, and C _1-4 halogen alkoxy;

R ⁴ and R ⁵ are each independently selected from:

(i) H;

(ii) halogen;

(iii) - (C _0-3 alkylene) C _3-10 cycloalkyl, wherein cycloalkyl is optionally substituted with 1 to 4 independently selected R ^f ;

(iv) - (C _0-3 alkylene) heterocyclyl containing from 3 to 10 ring atoms, and from 1 to 3

23,037,780 ring atoms each independently selected from N (R ^e ), O and S, where heterocyclyl is optionally substituted with 1 to 4 Rf;

(v) - (C _0-3 alkylene) (C ₆ -C ₁₀ aryl) optionally substituted with 1 to 4 R ^g ;

(vi) - (C _0-3 alkylene) heteroaryl having 5 to 10 ring atoms, with 1 to 4 ring atoms each independently selected from N, NHO and S, wherein the heteroaryl is optionally substituted with 1 to 3 R ^g ;

(vii) Cl _-6 alkyl optionally substituted with 1 to 2 independently selected R ^h ; and (viii) - (C _0-3 alkylene) C _4-10 cycloalkenyl, wherein cycloalkenyl is optionally substituted with 1 to 2 Rf;

R ⁷ and R ⁸ are H;

R ^a represents:

(i) Cl _-6 alkyl optionally substituted with 1 to 2 R ^h ; or (ii) - (C _0-3 alkylene) C _{3-1 0} cycloalkyl, wherein cycloalkyl is optionally substituted with 1 to 2 Rf;

(iii) - (C1 _-3 alkylene) heterocyclyl having from 3 to 10 ring atoms, in which from 1 to 3 ring atoms, each independently selected from N (R ^e ), O and S, where heterocyclyl is optionally substituted from 1 up to 4 independently selected R ^f ;

(iv) - (C _0-3 alkylene) phenyl optionally substituted with 1 to 4 independently selected R ^g ; or (v) - (C _0-3 alkylene) heteroaryl having 5 to 10 ring atoms, with 1 to 4 ring atoms each independently selected from N, N (R ^e ), O and S, where heteroaryl is optionally substituted with 1 to 3 independently selected R ^g ;

in each case, R ^c and R ^{d are} independently H or C _1-4 alkyl;

in each case, R ^{e is} independently H or Cl _-4 alkyl;

in each occurrence, Rf is independently Cl _-6 alkyl, Cl _-4 haloalkyl, —OH, F, Cl, Cl _-4 alkoxy, Cl _-4 haloalkoxy, cyano, or phenyl optionally substituted with 1 to 4 R ^g ;

in each case, R ^{g is} independently halogen, cyano, Cl _-6 alkyl,

Cl _-4 haloalkyl, Cl _-4 alkoxy or Cl _-4 haloalkoxy; and in each occurrence, R ^{h is} independently —OH, F, C _1-4 alkoxy, Cl _—4 haloalkoxy, or cyano.

In another aspect, a compound of the invention is selected from

- 24 037780

- 25 037780

- 26 037780

or a pharmaceutically acceptable salt thereof.

In another aspect, a compound of the invention is selected from

- 27 037780

- 28 037780

or a pharmaceutically acceptable salt thereof.

In another aspect, a compound of the invention is selected from

or a pharmaceutically acceptable salt thereof.

- 29 037780

In another aspect, a compound of the invention is selected from

or a pharmaceutically acceptable salt thereof.

In another aspect, a compound of the invention is

or a pharmaceutically acceptable salt thereof.

In another aspect, a compound of the invention is

or a pharmaceutically acceptable salt thereof.

In some aspects, R ¹ and R ² , together with the nitrogen atom to which each is attached, form a saturated or unsaturated ring having 5 to 6 ring atoms, said ring comprising: (a) 3 to 5 ring carbon atoms , each of which is optionally substituted with 1 to 2 independently selected R ^f , and (b) 0 to 1 ring heteroatom (in addition to the nitrogen atom attached to R ¹ and R ² ), which is independently selected from: N, N ( R ^e ), O and S; and with the proviso that one of the ring atoms is —C (O) -. In other aspects, R ¹ and R ² , together with the nitrogen atom to which each is attached, form a saturated or unsaturated ring having 5 to 6 ring atoms, said ring comprising: (a) 3 to 5 carbon ring atoms, each of which is optionally substituted with 1 to 2 independently selected R ^f , and (b) 0 to 1 ring heteroatom (in addition to the nitrogen atom attached to R ¹ and R ² ) which is independently selected from N, N (R ^e ), O and S; and with the proviso that one of the ring atoms is —C (O) -. In other aspects, R ¹ and R ² together with the nitrogen atom to which each is attached form a saturated or unsaturated ring of 5 to 6 ring atoms, wherein the ring comprises: (a) 3 to 5 ring carbon atoms, each of which is optionally substituted with 1 to 2 independently selected R ^f , and (b) 0 to 1 ring heteroatom (in addition to the nitrogen atom attached to R ¹ and R ² ), which is independently selected from: N, N (R ^e ) , O and S; and with the proviso that one of the ring atoms is —C (O) -. In other aspects, R ¹ and R ² together with the nitrogen atom to which each is attached form

In other aspects, R ¹ and R ² together with the nitrogen atom to which each is attached form

In another aspect, the present invention relates to a compound selected from the exemplary examples, or a pharmaceutically acceptable salt thereof.

In another aspect, the present invention relates to a compound selected from any subset of the list of compounds, or an individual compound, exemplified within the scope of any of the aforementioned aspects.

- 30 037780

One of ordinary skill in the art will understand that some of the chemical structures described herein may be represented on paper by one or more other resonance forms; or may exist in one or more other tautomeric forms, even when kinetically the skilled person recognizes that such tautomeric forms represent only a very small fraction of the sample of such compound (s). Such compounds are expressly included within the scope of this disclosure, although such resonance forms or tautomers are not explicitly represented herein.

In some embodiments, the implementation of the connection is selected from the compounds shown in table. 1 below. Biological assays used to test compounds are discussed in the examples section. The key to the ranges of activity: A = <1 μM; B => 1 μM, <20 μM; C => 20 μM, <100 μM; D => 100 μM.

Table 1

Structure Compound hNLRP3 ECso agonist (mkM) TLR7 ECso (mkM) TLR8 ECso (mkM) LCMS [M + HJ + СН ₃ Ί 1 V сМ HN'N 101 0.27 D D 308.3 H ₃ C CH Y 1 / ° 1 J ^N \ = N 102 1.60 D D 336.3 NH ₂ 103 0.52 D D 424.3

- 31 037780

СН ₃ <СНз ju JL / ⁰ JT ^H H ₃ c ^^ 104 0.60 D D 298.1 nh ₂ n ^ W4 ⁿ 4 ju Y / ⁰ f # HN ' ⁿ 105 0.69 348.2 Y ⁿ 2 CH _{3 n} A ^ n hn ^ ju JL ^x / ilT ^N 'JJ CH ₃ fT ^ HN' ⁿ 106 0.70 D D 322.1 H ₃ C nh ₂ Vch _{3 N} VH _N 4 JI Y / ⁰ <YN HN'N 107 0.64 350.3 A z-z X Z ω 1 ω = Ο o w T O ω 108 0.87 D D 386.3 CH ₃ CH ₃ Y ⁿ 2 (0 - (- CH _{3 Ν} Α <Ν CH3 1! Υ 7 “^ ⁰ g / T ΗΝ ' ^ν 109 0.88 D D 408.3 H ₃ with NH ₂ H ₃ C ^ n-CH ₃ ^N TvJ ^ n ΗΝ ' ^ν 110 0.90 D D 379.3

- 32 037780

^ CH ₃ ην ^ ji 2 'ζ ⁰ l ΗΝ' ^ν 111 0.72 D D 336.3 ΟΗ ₃ ^ ² <CH ₃ ji / L ⁰ ri ^ r ^N e / 'J HN' ⁿ 112 1.28 D D 350.1 Y ^H 2 CH ₃ ^H / ^N 4 ji Λ / ⁰ ίΐΎ ^ ^Ν hYG HN ' ⁿ 113 0.56 322.3 CH ₃ nh ₂ ( ³ ₀ Αν V fj н M HN ' ⁿ 114 1.04 D D 419.4 nh ₂ h ₃ s 0 n-S '° II I CH ₃ DA HN' ⁿ 115 0.90 D D 372.1 nh ₂ 0 _n A ^ n hn-s * ° H Г CH ₃ HN ' ⁿ 116 1.16 358.0 ΝΠ ₂ j! ( ^N and Lry ^CHs \ ^ N 117 1.26 C D 362.4

- 33 037780

CH ₃ IX nh ₂ ( ³ L> l JL / ⁰ ΗΝ ' ^Ν 118 1.36 D D 376.2 Α ζ-ζ “γΧ one ω = Ο ο W I Ο ω 119 1.38 D D 372.1 2 ^ Η ι ₃ 0 ^1/0 <Α ^'Χ ΗΝ' ^Ν 120 1.69 D D 362.2 SN ₃ Ν ^η 2 (CH3 NAV / A ii Λ ⁰ A CH3 Cr ^ ΗΝ ' ^ν 121 1.82 D D 364.2 νη ₂ Н ₃ С _СНз ν A η l 1 ⁰ ΗΝ ' ^ν 122 1.40 D D 350.2 n _{3 2} n ₃ νη with _l ^ _CHs ii ^1/0 ΗΝ'Ν 123 1.9 D D 376.2

- 34 037780

CO I about H «<Z) = O T 1 'А ZI CHA > = z 124 2.4 D D 386.2 CH ₃ (^ ch ₃ n \ va JAZ ^- ^ ⁰ rX HN ' ⁿ 125 2.1 D D 364.2 nh _2H3 c ^ О nV ^ 4 J! A / - ⁰ <YN (X HN ' ⁿ 126 2.1 412.4 CH ₃ nh _{^2/3 0} νΆ ^ν Α ^ν 4 January 11 7- hn-ch ₃ ^N X HN ^'n 128 2.9 D D 365.2 H ₃ C CH ₃ n \ v ⁿ A 1 A / ⁰ HN ' ⁿ 129 3.6 D D 336.2 _Z CH ₃ ΐ ^Η 2 <NH ₂ 1! JL 7 “^ ⁰ jlj \ = N 130 3.3 D D 351.2 nh _{2 n} An nh ₂ 1 A 'z fX hn-n 131 3.5 D D 280.3

- 35 037780

, ch ₃ G ² <pHz L A ⁰ ί T ⁿ \ = Ν 132 3.5 D D 350.9 ch ₃ <ζ-ΟΗ ν \ ^ν A 1 A / - ° Zy ^ N rN HN-N 133 3.5 366.3 CH ₃ CH ₃ (Q - (- CH _{3 n} xVn N-Λ CH3 l 1 t ^- ^ ° Zy ^ N ίι T ⁿ Vn 135 4.6 409.2 H ₃ NH ₂ C n ₃ C _x ³ \ _ _CHs l A / ⁰ eN HN ^'n 136 4.8 D D 362.2 nh ₂ oVyV N'S'V ^^ 1 L L 'HN-N 137 5.4 424.4 ° A Z ^x ΖΞ CM) ----- ('Yae Oh 138 6,7 C D 373.2 ^z \ o =% YY I / ------ \ ZZ I о JI O ^ o ΞΕ w 140 7.3 D D 364.2

- 36 037780

/ ---- \ ^m ъ 141 9.0 D D 316.1 ΝΗ _of H ₂ gc _CH ^Ν ^ 4Ν / Ν ~ ^ ^removal of JI / ⁰ AAn hn-n 142 9.1 D D 380.2 nh ₂ ii V> _ jj ^H Cl 143 9.4 C D 316.1 o ^z O Ά 153 12.0 C D 393.4 7 ^H 2 CH ₃ Ad ^H / ^N ^ Л A / - / YES ί J ^H H ₃ CO ^^ 154 16.1 D C 272.0 H ₃ C CH ₃ NH ₂ h ₃ C - y p- \ ³ N - '' H-L / ^N V ^CH 3 L X / ⁰ f T ^H V- NH 155 17.4 D D 408.2 nh ₂ JI | ____ ^{/ N} <3 j (jH ъ 157 23.4 C C 398.2

- 37 037780

CH ₃ <CH3 Η ₃ Ο (Τ ^ 158 24.5 D D 314.3 ch ₃ ^ 2 (CHs. A CI 159 25.9 D D 352.0 nh ₂ СН 1 СН ₃ Ν ^ γ-Ν НМ-2 Λ 162 42.0 D D 308.3 J ¹ about x 163 48.4 C D 350.2 Y < ^C \ n, Ανί ^^ OSN ₃ 164 4.6 D D 384.2 CH ₃ NH ₂ (CH ₃ ιι A ⁰ <N ^ n 165 0.19 D D 360.2 / = Z О tx _z A // т "x \ Z о II ω О - ^ О I ω 166 3.4 D D 361.2

- 38 037780

CH ₃ A ² (CH ₃ 1! A z ^- ^ ^{0 n} 167 58 D D 361.2 CH ₃ N ^h 2 (CH ₃ 1 A / ⁰ hr Ay ^NM 3> 1 H n ^ AAA yr H ₃ c 168 101 D D 378.2 CH, (CH ₃ 1 A ⁰ H ₃ C- / X 169 92 D D 392.2 ch ₃ ^ ² (CH ₃ l X / ⁰ AAA ^ N HI ^H h ₃ cAX CH ₃ 170 7.9 C D 326.2 CH ₃ nh ₂ (CH ₃ CH ₃ 171 3.2 IN D 312.2 ph ₃ (, CH ₃ N '^ W ^ ^N ^ Ji A / ⁰ / U ^H HsC ^ 172 2.1 IN D 326.2 7 ^Ng (ph ₃ j ^ X / ^{0 xi n} 173 0.84 D D 352.2

- 39 037780

CH ₃ 7 ^Н 2 (СН ₃ l JL / ⁰ VU ⁿ Μ 175 2.6 D D 379.2 CH ₃ ^ ^Η 2 (CH ₃ n \ ^N va ^N 4 1! A 7 ^- ^ ° U ^H 176 25.7 D D 284.1 NH ² ( ³ CH ₃ m \\ 7Л ii A / ⁰ AJ ^H Br ^^^ 177 7.4 D D 362.1 CH ₃ (CH ₃ n \ va JA / ⁰ AA ^ N ί TH Cl 179 12.5 D D 318.1 CH ₃ NH ² (CH ₃ ⁰ 180 1.5 D D 350.2 CH ₃ nh ₂ (CH ₃ JI A / X ⁰ jfjP h ^CH3 Br ^^ 182 5.9 D 376.1 ΝΠ ₂ ( ^З SN ₃ n ^ Wv ^N A ji JL / ^{0 H} 183 0.52 D D 350.3

- 40 037780

Y ^H 2 Н ₃ С СНз 187 64 IN FROM 270.2 NH ₂ n ₃ С СНз •? С base ₃ 188 5.7 D D 370.2 ^ ^H 2 ( ³ CH ₃ nUijU l Jl / ⁰ / V'N JU and Yy ^ ° ^CH 3 190 10.3 D D 444.4 .СНз nh ₂ (СНз Ν Η oU H ₃ U 191 65 D D 441.2 , ch ₃ 7 ² <ph ₃ Br 194 7.9 D D 362.1 Y ² (En ₃ irVY uu \ Y SN ₃ 196 32,7 D D 488.2 , ch ₃ νη ₂ (ch ₃ Cl 197 13.7 D D 318.0

- 41 037780

, ch ₃ D ² (CH ₃ Di 0 ^ 0 ^ CHs 198 24.2 D D 434.2 СН ₃ Y ^H2 (СН ₃ n \ ⁿ X4 ii X / ⁰ XXn US С ° СН ₃ 199 D D 369.3 CH ₃ ^ ^H 2 (o-sn ₃ Br 200 And, 9 D D 378.0 OT — ά 'Ηλτ \ (ΙΌ Ή f s ω O'll'o About t 201 20.8 D D 398.2 CH ₃ nh ₂ (p nVv / ^ ¢ 0 Q 1 CH ₃ Br 202 35.9 D D 474.1 ph _{3 ΝΗ 2} (0 n \ V v 1 X / / ^СН s | f ^ | n ^НзС Br 203 10.8 D D 390.4 ph ₃ o Bg 204 6,7 D D 424.0

- 42 037780

СН ₃ СН ₃ У ^Н 2 (О - (- СН ₃ (У С Λ ~ ν \ о V_ / ° 205 36 D D 483.4 co I o £ \ ^ 0 o Γ 1 - 7 ^x m 2 - ~ / ^z aK CX 207 73 D D 350.1 ΝΗ2 (Рнз РтХЧ FJ V Q СН ₃ 209 22 D D 530.2 νη ₂ CH ₃ (CH ₃ ώΧ wow HN-ch ₃ 210 61 D D 463.2 CH ₃ CH ₃ nh ₂ (o - (- SN ₃ ^CH 3 and S V η / ^SN 3 0 ^ ^Ν \ A ^ CH ^{3 0} SN, 211 40 D D 582.4 ph _{3 ΝΗ 2} (CH ₃ lx \ _ / ^CHs 212 85 D D 524.4 ΝΗ ₂ ( ^З SN ₃ ij S Ap, ^N ~ CH ₃ Н ₃ С 213 17.6 D D 477.2

- 43 037780

ph ₃ nh ₂ (ch ₃ l X / ° XT 4 Br>"\ z ° 0 214 51.5 D D 517.2 nh ₂ Q n ^ V ^N \ ^ ^N ъ XJ 4 Br- ^ ⁴ ^ \ _zP, ^N ~ CH ₃ H ₃ C ³ 215 18.2 D D 473.1 o% 'ft) _o l? o ° I 216 6.6 D D 416.2 ^ ^H 2 (ch ₃ ji A / ⁰ XI \ _f HN-cH3 217 96 D D 417.2 PH ₃ T ² <CH ₃ A "vL ^ ^ \ ^ P ° ~ CH ₃ 218 1,2 D D 464.2 ph ₃ i ^ ^H ₂ (ch ₃ J! X / ⁰ XJ h CV '''' ^ \ __ zP ° ~ CH ₃ 219 2.0 D D 418.2 TY v ^z ^ o 220 51 D D 473.2

- 44 037780

^ ’. СНз ryVA jj h Cl \ __ £ oh Ν SNz 221 85 D D 486.4 , SN ₃ NH ₂ <CH ₃ ji Y / ° 1 J γ 222 42 D D 457.2 .СНз СНз NH ² (О ~ (“СН ₃ СНз ά \ 1> О 223 77 FROM D 531.3 U ⁿ 2 SNz X T ⁿ 225 10.9 D IN 321.9 co with X co I O X ° V / ° co b _o δ Chk / chy o co LL 226 85 D D 509.4 Y ⁿ 2 CH _{3 s} 4l ην-> 1 X / IT γ F ₃ C ^^ O-CHs 227 22.7 FROM D 410.3 CH ₃ CH ₃ (0 - (- CH ₃ ^CH3 .eV \ o 0 228 93 D D 565.3

- 45 037780

ch ₃ nh ₂ (ch ₃ l X / ⁰ U h F ₃ C ^^ o-ch ₃ 229 12.3 D D 452.2 t about about about L __R 230 4.5 D D 388.2 U ⁿ 2 CH ₃ H ⁴ 233 120 Β D 256.1 Oh X 278 10.8 C D 282.1 _{νη 2} ζΧ ii HH Jf T ^H 279 7.5 D D 359.1 g o n chu- Ν / ^Ν Ά Ρ 280 0.22 D D 358.2 _{νη 2} ζχ ϊι jA__ ^{/ Ν} u 4 ^Β 281 1.4 D D 392.1

- 46 037780

nh ₂ 4 jA_ ^{/ N} s 282 4.8 D D 372.2 Y Z ^ ZI co \ 283 6.5 D D 388.2 O ^"m / ---- ^(w IZ, ZA 284 0.86 D D 376.1 __ _z o Y Ae AA o 285 5.7 D D 392.1 I about VA 'φ-χ'ί X P 286 0.61 D D 372.2 ΞΕ ABOUT VA 'C 287 1.08 D D 388.2 -Z. about VA IZ, z P 288 10.1 D D 383.1

- 47 037780

nh ₂ 1! ^Νχ > ___ ^{/ N} yA 289 1.58 D D 376.1 ₀ f ° 1 Z ^X ZI CM) ----- (ΛΑ Th t 291 oh and D D 372.2 Ltd ) ----- (th X P 292 4.48 D D 388.2 'Ά m 294 4.6 FROM D 360.0 NH ₂ AV ⁴ o L J ⁿ o \ = N 295 1.20 D D 348.1 __o A, cm) ( Z-Z V about with T 296 5.1 D D 362.2 νπ ₂ Ϊ jTV ^ ¹ u χΡΑν ^Η ° i 298 21 IN D 366.2

- 48 037780

nh ₂ Ν'η '' ^N \ ^ __ / ^N 43 1 J ⁿ Vn H ₃ C 301 2.93 D D 362.2 nh ₂ ϊ jU_ <Y ^NH 302 0.93 D D 364.2 □□ / ---- ( ^M IZ _x z O ^ Z ^ \ I 1 / <y ~ ^ 303 ID FROM C 373.8 nh ₂ N __ / ^N 4e P ⁿ Λ J НзСО '^^ 304 36.4 389.2 νη ₂ Л and F rV ^ N 1 JL J ^H u 307 12.3 377.1 c ^ ° Λ Z ^ ZI о ω ^χ 308 1.06 364.1 C <z> chi “Y 309 0.24 D D 364.1

- 49 037780

__ / About cm) ( about <5 315 1.18 D D 383.2 from Hell Ά-Α 316 ι, ιο D D 436.1 nh ₂ 0СН ₃ AAfj _М ^ ДД and 319 6.2 D D 389.2 Г ² О ϊ jrV ^ ^N l JAT n _Ν ΑγΑ / and 320 1.5 D D 393.1 r = -Z. W C>) = \ > A Ά 321 4.5 C D 393.1 P with ABOUT 5 = 2 W d 324 6,7 D D 427.1 nh ₂ ϊ • e rAf N L J ⁿ Y CN 326 2.7 D D 384.1

- 50 037780

__, Ο CXI) ------- ( about 327 13 D D 384.1 Г ² О JJ _Яс <Υ N XJ ⁿ Υ Cl 328 4.1 D D 393.1 νη ₂ А ϊ if V ^ ^N jO "А 330 1.6 D D 377.1 νη ₂ (А ϊ jA_ ^{/ Ν} u χ τ ^Η τ F 331 3.2 D D 377.1 ^σ 1 ζ ^ ζχ h 332 3.3 D D 378.2 1 νη ₂ Α η (Ύ ^ ^Ν ό rAg ^Ν XX ^Η and 333 1.9 D D 435.2 g _ο:? with OCH ₃ 335 6.5 D D 382.2

- 51 037780

_{νη 2} ϊι jT V Xj Υ u X ' ^CH s H3C 337 80 D D 429.2 νη ₂ ^ Χ ν ητ ^Ν ^ χ ^Ν b ij γ Cx h ° ^Ν / N ~ CH ₃ H3C 340 fifty D D 472.4 νη ₂ у ϊ jx) __ ^{/ Ν} 'e ij γ ОНЗ 341 2.1 D D 462.2 νη ₂ у Ar γ νΧΧ ^^ V /, ° OCH ₃ 342 3.3 D D 459.3 Ν ηΧχΧ A and y 0 345 64 453.2 r O ltHC 346 17.3 D D 296.1 r about OH Oso 347 41 D D 396.2

- 52 037780

Ψ ^Η 2 H ₃ C N-CH3 j JL XX ^ N / X ΗΝ- ^ν 348 0.89 D D 308.0 νη ₂ n ₃ s sn ₃ Αν l X / XXn ΗΝ ' ^Ν 349 0.60 D D 322.1 CH ₃ / —N NH ₂ (\ l Y / ⁰ χχχ X HN ' ⁿ 350 10 377.2 / -0 nh ₂ (\ l Y / ⁰ XXn X ± HN ^N 351 2.17 364.2 nh ₂ n ^ WX a л X / ⁰ XX ^N V HN ' ⁿ 352 2.23 362.1 NH ₂ O = X'X nVX ^J Xn ' ^A ^ Vn (R) -enantiomer 357 0.64 D D 424.4

- 53 037780

nh ₂ J! 1 z ^- ^ i T ^H Vn (8) -enantiomer 358 0.63 D D 424.4 nh ₂ ° f ° d AL ^ O ⁿ hn-n 363 5.8 D D 426.4 NH ₂ O = Z °> ήν ft HN ' ⁿ 365 5.0 D D 350.2 CH ₃ N nh ₂ 0 = / 7 ϊ ttV ^ ¹ ftHN-N 368 3.9 D D 363.2 nh ₂ H ₃ c ^ oh mA ^ n n-4 11 1 φ— ⁰ ft HN ' ^N 370 4.8 D D 352.3 nh ₂ h ₃ c ^ ^nh 2 r / t HN- ⁿ 372 8.3 D D 351.3

- 54 037780

H ₃ C nh ₂ H ₃ c N-CH ₃ X ^- HN ' ⁿ 375 2.4 D D 365.2 NH ₂ H ₃ C j \ ^N p ъ HN ' ^N 376 3.6 D D 398.3 A z 'ZI IN) ( CHU Xz M 377 4.3 D D 405.3 ^T y ° fl A / ^ZT Λ = ^ζ \ NX: z 379 2.0 D D 336.3 CH ₃ nh _2H3 c ^ _Λ ν 'n ³ X HN- ^N 383 3.6 D D 393.4 CH ₃ -CH ₃ NH ₂ ζ p - (- CH ₃ ApA ^CH3 nn H ₃ C 393 7, H D D 422.2 n T co X О X one zx CNJ) ------ ( 'Yae 397 1.05 D D 424.4

- 55 037780

nh ₂ ch p 398 1.22 D D 308.3 « ^N 2 CH ₃ N Jj ^H V 399 1.06 D D 324.2 nh ₂ ch ₃ Ayr ⁷ riY N s- ^ 400 1.79 D D 324.2 nh _{2 n} AL-N HN-СНз j JL X 'HN'N 401 1.23 D D 294.3 nh _{2 n} 4-N HN-CHz l JL / ⁷ ί ϊ n 402 1.31 D D 294.3 NH ₂ ^ N ' ^CH 3 Xv ⁿ⁴ ° ι / j X X ^ PoCH ₃ HN'N 403 3.66 D D 483.3 9 NH ₂ A ^N V ⁴ o РРосн ₃ HN ' ⁿ 404 2.50 D D 590.3

- 56 037780

7 ^Н 2 Н ₃ С р-СНз prVA ΑγΝ rA ΗΝ'Ν 405 2.19 D D 352.3 SN ₃ ΝΗ ₂ (O-SNz Ν ^ Ν ^N p γΥ ΗΝ ' ^Ν 406 1.37 D D 366.1 ο _{νη 2} ρρ ΗΝ ' ^Ν (I) -enantiomer 407 6.55 D D 424.2 0 ~ (S) Г ² Ο ρβ_ ^{/ Ν} and fAr Ν Λ ΗΝ ' ^ν (8) -enantiomer 408 5.23 D D 424.2 νη ₂ V / ^ OCHs ΗΝ ' ^Ν 409 2.36 D D 546.5 Ν — l νη ₂ ¢ / Ά ΛΑ Αζ ^basic ΗΝ ' ^ν 414 3.83 D D 479.4

- 57 037780

СНзГ ° \ nh ₂ ( ³ V Λ ι Λ ⁰ Α 'ην-ν 417 7.4 D D 392.4 CO co Ά Az ο σ> 419 0.91 D D 366.3 СНз η ₂ ν <ρ Α_Ν Ν — 9 ^Η 3 Эо — рсн ₃ ΑΑν СНз _N JU ^Ьн > HN Л 420 0.19 D D 422.4 ΝΗ GN ₂ 1 ⁿ s l nA-N HN- (L. P Y ^/ CH ₃ nJU ^h hn'JT 421 0.24 322.0 p H ₂ NH ₃ C- (CH ₃ zVx A НзС ^^ 422 0.28 > 10 > 10 374.1 NH ₂ A) N ^ yA4 / xAn JU ^H Vn 423 0.62 D D co T co О co = c I T o-ko ° <s / ° A Z ^ ZI CN) < 424 0.79 D 424.2

- 58 037780

p nh ₂ H ₃ cY ch ₃ 1¼ V <Y ^N Vs 425 0.44 D 366.2 Re about 426 4.3 D p n ₃ s — 0 CH ₃ ί T ^H T F 427 5.0 D D 379.0 ΝΠ ₂ ⁿ Xa / ^N v J! JL z ~ ^{J 0} / VP XT h 428 5.1 D nh ₂ cw °> ^N | T ^N ^ _y H 429 5.8 D 350.2 0 _2N NH ₃ ^ C (CH ^{_{mA ^ n n- N |? T}} \\ __ / fj X ^{CH Br} CH ₃ 430 7.45 D > 10 418.1 p NH ₂ H ₃ c4 / ^СН s N ^ ryY _Bf jCO q ^z CH ₃ 431 8.67 D > 10 434.1

- 59 037780

CH ₃ NH ₂ (\ θ ί J ^H nXA HNJ 432 ~ 10 377.2 Η ₃ Χ ^Η 3 ° Do ^ CH ₃ l ² O ΑΧ JL J ^H M 433 11.5 474.3 nh ₂ X _w A ^ n nX n X \> _ / 0 Z \ X X ^H 434 15.5 > 30 > 30 nh ₂ 0 = ( ³ CH ₃ nX ⁿ x Ay n X? CH ₃ 435 19.3 D 404.2 CH ₃ CH ₃ nh ₂ ( ³ ₀ V ³ nVx ⁿ ^ l Y / ⁰ XX N X HN ' ⁿ 436 1,3 > 50 > 50 390.0 Λ- <Ύ II T ^x XO CH ₃ / УУ ^Н HN ' ⁿ 437 1.1 D D 440.4 nh ₂ n-S'N ^N X JI Y / - ⁰ XX N X HN'N 438 4.2 D D 413.0

- 60 037780

CH ₃ 55 NH ₂ <) = N L A / ⁰ <YN HN ' ⁿ 439 5.7 D D 413.2 ch ₃ OH nh ₂ / \ / \ UHo Ava ^сНз J! JL / ⁰ r /> hn- ⁿ 440 18.4 D D 408.4 OCH ₃ n \ v4 Л A / ⁰ ГГ HN ' ⁿ 441 0.69 D > 62 442.0 N ^ \ CH ₃ (/ A nh ₂ ( ³ W nV ^N ^ H _l L A / ⁰ N c / d HN ' ⁿ 442 2.6 D D 414.0 CH ₃ / = \ Γ ² <1! A ° X> s ^ N лА - HN- ^N 443 0.86 D > 62 426.0 nh ₂ p λΏ mALn nA HN-N 444 0.93 D IN 454.1 H ₃ c ₄ Ych ₃ nh ₂ ^ ^Нз \ ^N 4 Л A ⁰ HN- ⁿ 445 0.74 D > 62 406.1

- 61 037780

СН ₃ Λ νη ₂ / Α _Νχ Ad A ^СНз II Υ / ⁰ ΑΑν Α ΗΝ ' ^ν 446 0.67 D D 415.2 CH _S Ρ ΝΗ ₂ ζ ³ Τ 'ι JL / ⁰ ΑΑ Α ΗΝ' ^ν 447 1.5 D Β 440.2 _ζ % ΝΗ ₂ F ^h 'Ρ ΑαΝ Α ΗΝ-Ν 448 2.0 D D 419.3 α ΡΡ Vw4 ^Cl l Ρ / ⁰ ΑΑΝ Α ΗΝ-Ν 449 0.90 D C 446.0 Λ ^- Ν CH ₃ / νη ₂ ζ) = / νΑ ^ ν ^ν Α 1 Υ / ⁰ ν Α ην- ^ν 450 4.5 D D 413.0 SNz ^ΝΗ 2 / ^SNZ νΆΟλ 1 Ρ 7— ° AkhD-m Α ην- ^ν 451 1,3 D C 465.2 ΝΗ, ha J! Ρ Α ° AAA Α ^ ^Η ην- ^ν 452 1,2 D > 62 446.0

- 62 037780

SN ₃ nh ₂ ( ³ / А ji A / ^СНз - HN ' ^N 453 0.71 D D 364.1 CH ₃ CH ₃ <CH ₃ nh ₂ ζ ³ W ³ J! A / ° A HN ' ⁿ 454 0.74 D D 407.1 SNZP nh ₂ ζ ³ nA ^ n ^N 4 II А A ^ о A HN ' ⁿ 455 2.0 D D 421.4 NH F ^Hg / nh ^CH _2; HN ^ _{CH3 n} A ^ n ch ₃ 11 A / A o aaa A HN ' ⁿ 456 2.2 D D 407.1 CH ₃ CH ₃ NH ₂ (HN ^ J! A / ⁰ A HN ' ⁿ 457 6.6 D D 379.3 CH ₃ NH _{2/0 n A ^ n n-S '} ° 1 L - N-CH ₃ ^CIP Cj H'HN'N 458 2.1 D D 415.0 CH ₃ N ^h 2/0 „\ IDO n4 <n ns; Ϊ X ^CH 3 fjH ^НзС HN-N 459 6,7 D D 413.9

- 63 037780

СН ₃ nh ₂ Г сэ \ сО X “HN ' ^N 460 1.5 D > 62 414.0 N ^h 2 ch ₃ ^{NAyy N- /} eA? N-NH he 462 5.5 D D 352.3 nh ₂ О = ХА ___ ^{/ Ν} / UU> H ₃ c Vs 463 0.22 D D 392.2 νη ₂ 1 Xs νΧ-Ν ηι \ Χ ЛчГ ρ fj a Ζ ^ Γ ^ Xn ₃ ν-νη 464 0.24 D D 336.3 _{νη 2} ΟγΧ m_ ^{/ Ν} JU> n ₃ s Χν 465 1.0 D D 376.2 _{νη 2} ΟγΧ J) __ ^{/ Ν} / UU> ^H 3 ^C Vnh 466 1.1 D D 376.2 nh ₂ A 7 ^CH 3 ^N fj V Vh ₃ Xn 467 1,2 D D 336.3

- 64 037780

nh ₂ 1 P ⁿ s hn ^ Al P GT \ 468 2.1 D D 352.1 Xr CM) I NE mJ 469 0.67 D D 408.0 CH ₃ NH ₂ (^ CH ₃ mX ^ n nA; t V ° AAn AA O 'oh 470 0.69 D D 424.1 T ² pH, m> = A - N HN— ⁷ x LA- ⁷ iiT ^N оД ^ OH 471 1,2 D D 368.1 0 nh ₂ H ₃ c ^ _CH3 ⁿ Spirit ^-7 V ^ CH3 SU 472 0.43 D D 393.9 NH / ^CH 3] ^H2 (^ CH ₃ N ^ VN nA A IL / ^{--7 0} D1 V / hAA> SN ₃ Vnh 473 0.22 D D 392.3 0 ^ ^Η 2 H ₃ cX CH ₃ rr ^N X 1 J Ah Vnh 474 1,2 D D 378.3

- 65 037780

nh, LHn 1 \ / ^ CH ₃ l 1 / - ⁷ o N-NH OH 475 23 D D 408.1 о НзСЯ СН ₃ о А? \ = N OH 476 6.1 D D 394.1 Ϊ ^Η * RN, oh Huh? 477 6.3 D D 352.1 A ZZ Yx \ _ / X / ---- ( ^m IX. Л ^ω θ '] о □: W 478 3.7 D D 408.1 A Ζ — Ζ / --- \ ^mu t co 479 0.67 D D 378.0 with X ABOUT co 1 ^ ° HA £ A Uz 480 1.9 D D 408.0 CH ₃ Η ₂ Ν (^ CH ₃ Ν ψ - / 0 ί Τ Jan ₃ Ρ VN 481 0.63 D D 392.2 CO A τ ^Τ \ / ^z ^ o CM V / χ Λα ^ζ Par 482 0.43 D D 410.1

and their pharmaceutically acceptable salts.

Pharmaceutical compositions and routes of administration.

General information.

In some embodiments, a chemical (eg, a compound that modulates (eg, agonist or partial agonist) NLRP3, or a pharmaceutically acceptable salt and / or hydrate and / or co-crystal and / or drug combination) is administered as a pharmaceutical composition that includes a chemical and one or more pharmaceutically acceptable excipients and, optionally, one or more additional therapeutic agents, as described herein.

In some embodiments, the pharmaceutical composition comprises a compound according to

- 66 037780 of the present invention or a salt thereof and one or more pharmaceutically acceptable excipients. In some 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 some 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.

In some embodiments, the implementation of the chemicals can be entered 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 tweens, poloxamers or other similar polymeric delivery matrices, whey proteins such as human serum albumin, buffering agents such as phosphates, tris, glycine, sorbic acid, potassium sorbate, partial glyceride mixtures of saturated vegetable fatty acids, water, salts or electrolytes such as protamine sulfate, disodium hydrogen phosphate, potassium hydrogen phosphate, sodium chloride, zinc salts, colloidal silicon dioxide, magnesium trisilicate, polyvinyl pyrrolidone, cellulose-based substances, polyethylene glycol, sodium carboxymethyl cellulose, polyacrylates, waxes, polyethylene polymers and wool polymers. Cyclodextrins, such as α-, β- and γ-cyclodextrin, or chemically modified derivatives such as hydroxyalkylcyclodextrins, including 2- and 3-hydroxypropyl cyclodextrins, or other solubilized derivatives, can also be used to enhance delivery of the compounds described herein. document. Can be prepared dosage forms or compositions containing a chemical, as described herein, in the range from 0.005 to 100% with a balance composed of non-toxic excipient. The contemplated compositions may contain from 0.001 to 100% of the chemical provided herein, in one embodiment from 0.1 to 95%, in another embodiment from 75 to 85%, in another embodiment from 20 to 80% ... The actual methods for preparing such dosage forms are known or will be apparent to those skilled in the art; for example, see Remington:. The Science and Practice of Pharmacy, 22 ^{nd Edition (Pharmaceutical Press, London,} UK, 2012).

Routes of administration and components of the composition.

In some embodiments, these chemicals described herein, or a pharmaceutical composition thereof, can be administered to a subject in need thereof by any conventional route of administration. Acceptable routes of administration include, but are not limited to, buccal, dermal, endocervical, endosinusial, endotracheal, enteral, epidural, interstitial, intra-abdominal, or intra-abdominal, intra-arterial, intrabronchial, intrauterine, intra-duodenal, intra-coronal, intra-dermal, intra-duodenal intraepidermal, intraesophageal, intragastric, intraesophageal, intradural, intralymphatic, intramedullary, intrameningeal, intramuscular, intraovarial, intraperitoneal, intraprostatic, intrapulmonary, intrasinal, intravascular, intravascular, intravascular, intravascular, intravascular oral, parenteral, percutaneous, epidural, rectal, respiratory (inhalation), subcutaneous, sublingual, submucous (under the tissue layer located on from the mucous membrane), topical or local, transdermal, transmucous, transtracheal, ureteral, urethral and vaginal. In some embodiments, the preferred route of administration is parenteral (eg, intratumoral). In certain embodiments, the preferred route of administration is systemic.

The compositions can be formulated for parenteral administration, for example formulated for injection by the intravenous, intramuscular, subcutaneous, or even intraperitoneal route. Typically, such compositions can be prepared as injectables, either as liquid solutions or suspensions; also can be obtained solid forms suitable for use for the preparation of solutions or suspensions after adding a liquid before injection; and preparations can also be emulsified. The preparation of such formulations will be known to those skilled in the art in light of the present disclosure.

Pharmaceutical dosage forms suitable for injection use include sterile aqueous solutions or dispersions; formulations including 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 dosage form must be sterile and must be fluid to the extent that it can be easily administered. It must also be stable under the conditions of manufacture and storage and must be protected from the contaminating action of microorganisms such as bacteria and fungi.

The carrier can also be a solvent or dispersion medium containing, for example, water, ethanol, polyol (eg, glycerol, propylene glycol and liquid polyethylene glycol and the like), suitable mixtures thereof, and vegetable oils. The proper fluidity can be maintained, for example, by the use of a coating such as lecithin, by maintaining the desired particle size in the case of dispersion, and by the use of surfactants. Prevention of the action of microorganisms can be provided by various antibacterial and antifungal agents, for example, parabens, chlorobutanol, phenol, sorbic acid, thimerosal, and the like. In many cases it will be preferable to include isotonic agents such as sugars or sodium chloride. Prolonged absorption of the injectable compositions can be achieved by the use in the compositions of agents that delay absorption, for example, aluminum monostearate and gelatin.

Sterile injectable solutions are prepared by incorporating the active compounds in the required amount in an appropriate solvent with various other ingredients listed above, as needed, followed by filtration sterilization. Typically, dispersions are prepared by incorporating various sterilized active ingredients into a sterile carrier that contains a basic dispersion medium and the required other ingredients from those listed above. In the case of sterile powders for the preparation of sterile injectable solutions, the preferred methods of preparation are vacuum drying and freeze drying methods, which yield the powder of the active ingredient plus any additional desired ingredient from its previously sterile filtered solution.

Intratumoral injections are discussed, for example, in 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).

Pharmacologically acceptable excipients suitable for use in a rectal composition as a gel, cream, enema or rectal suppository include, without limitation, any one or more of the following: cocoa butter glycerides, synthetic polymers such as polyvinylpyrrolidone, PEG (for example containing PEG ointments), glycerin, glycerol gelatin, hydrogenated vegetable oils, poloxamers, mixtures of polyethylene glycols of various molecular weights and fatty acid esters with polyethylene glycol, petrolatum, anhydrous lanolin, shark liver oil, sodium saccharinate, menthol, sweet almond oil, sorbitol, sodium benzoate, SBN anoxide, vanilla essential oil, aerosol, parabens in phenoxyethanol, sodium methyl p-hydroxybenzoate, sodium propyl p-hydroxybenzoate, diethylamine, carbomers, carbopol, methyloxybenzoate, macrogol cetostearyl ether, cocoyl capryropyne alcohol, isopropyl alcohol, isopropyl alcohol liquid paraffin, xanthan gum, carboxymetabisulfite, edetate on sodium, sodium benzoate, potassium metabisulfite, grapefruit seed extract, methylsulfonylmethane (MSM), lactic acid, glycine, vitamins such as vitamins A and E, and potassium acetate.

In certain embodiments, suppositories can be prepared by mixing the chemicals described herein with suitable non-irritating excipients or carriers such as cocoa butter, polyethylene glycol, or suppository wax that are solid at ambient temperature but liquid at body temperature. and therefore melt in the rectum and release the active compound. In other embodiments, the compositions for rectal administration are in the form of an enema.

In other embodiments, the compounds described herein, or pharmaceutical compositions thereof, are suitable for local delivery to the digestive or gastrointestinal tract by oral administration (eg, solid or liquid dosage forms).

Solid dosage forms for oral administration include capsules, tablets, pills, powders, and granules. In such solid dosage forms, the chemical is mixed with one or more pharmaceutically acceptable excipients such as sodium citrate or dicalcium phosphate and / or: a) fillers such as starches, lactose, sucrose, glucose, mannitol and silicic acid, b) binders such as, for example, carboxymethyl cellulose, alginates, gelatin, polyvinylpyrrolidinone, sucrose and acacia, c) humectants such as glycerin, d) disintegrants such as agar agar, calcium carbonate, potato or tapioca starch, alginic acid, certain silicates, and sodium carbonate, e) dissolution retardants, such as paraffin, f) absorption accelerators, such as quaternary ammonium compounds, g) wetting agents, such as, for example, cetyl alcohol and glycerol monostearate, h) absorbents, such as kaolin and bentonite clay and i) lubricants such as talc, calcium stearate, magnesium stearate, solid polyethylene englycols, sodium lauryl sulfate and mixtures thereof. In the case of capsules, tablets and pills, the dosage form may contain buffering agents. Solid compositions of a similar type can be used as fillers in soft and

- 68,037,780 hard gelatin capsules using excipients such as lactose or milk sugar, as well as high molecular weight polyethylene glycols and the like.

In one embodiment, the compositions will take the form of a unit dosage form such as a pill or tablet, and thus the composition may contain, along with the chemical provided herein, a diluent such as lactose, sucrose, dicalcium phosphate, or the like; a lubricant such as magnesium stearate or the like; and a binder such as starch, acacia gum, polyvinylpyrrolidine, gelatin, cellulose, cellulose derivatives, or the like. In another solid dosage form, the powder, marum, solution or suspension (for example, in propylene carbonate, vegetable oils, PEG, poloxamer 124 or triglycerides) is enclosed in a capsule (gelatin or cellulose base). Also provided are unit dosage forms in which one or more chemicals provided herein or additional active agents are physically separated; for example, capsules with granules (or tablets in a capsule) of each drug; bilayer tablets; two-piece gel caps, etc. Enteric coated or delayed release oral dosage forms are also contemplated.

Other physiologically acceptable compounds include wetting agents, emulsifying agents, dispersing agents or preservatives, which are especially useful for preventing the growth or action of microorganisms. Various preservatives are well known and include, for example, phenol and ascorbic acid.

In certain embodiments, the excipients are sterile and generally do not contain unwanted substances. These compositions can be sterilized by conventional, well known sterilization techniques. For various excipients of oral dosage forms such as tablets and capsules, sterility is not required. The USP / NF standard is usually sufficient.

In certain embodiments, solid oral dosage forms may further comprise one or more components that chemically and / or structurally predispose the composition to deliver the chemical to the stomach or lower gastrointestinal tract; for example, the ascending colon and / or the transverse colon and / or the distal colon and / or the small intestine. Exemplary compilation methods are described, for example, in Filipski K.J. et al., Current Topics in Medicinal Chemistry, 2013, 13, 776-802, which is incorporated herein by reference in its entirety.

Examples include methods for targeting the upper gastrointestinal tract, such as Accordion Pill (Intec Pharma), floating capsules, and materials capable of adhering to mucosal walls.

Other examples include methods of targeting the lower gastrointestinal tract. Several enteric / pH sensitive coatings and fillers are available to target different areas of the intestinal tract. These materials are typically polymers that are designed to dissolve or degrade at specific pH ranges based on the region of the gastrointestinal tract in which drug release is desired. These materials also function to protect acid labile drugs from gastric fluid or to limit exposure in cases where the active ingredient may cause irritation of the upper gastrointestinal tract (e.g. hydroxypropyl methylcellulose phthalate series, Coateric (polyvinyl acetate phthalate), cellulose acetate phthalate , hydroxypropyl cellulose, methyl cellulose acetate succinate, Eudragit series (copolymers of methacrylic acid and methyl methacrylate) and Marcoat). Other methods include dosage forms that respond to local flora in the gastrointestinal tract, a controlled pressure capsule for colon delivery, and Pulsincap.

Ophthalmic compositions may include, but are not limited to, one or more of the following: viscosity-increasing agents (eg, carboxymethylcellulose, glycerin, polyvinylpyrrolidone, polyethylene glycol); stabilizers (eg Pluronic (triblock copolymers), cyclodextrins); preservatives (e.g. benzalkonium chloride, EDTA, SofZia (boric acid, propylene glycol, sorbitol and zinc chloride; Alcon Laboratories, Inc.), Purite (stabilized oxychloro complex; Allergan, Inc.)).

Topical compositions can include ointments and creams. Ointments are semi-solid preparations that are usually based on petroleum jelly or other petroleum products. Creams containing the selected active agent are usually viscous liquid or semi-solid emulsions, often either 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 oily phase, also sometimes called the internal phase, usually consists of petrolatum and a fatty alcohol such as cetyl or stearyl alcohol; the aqueous phase usually, although not necessarily, exceeds the oil phase in volume and usually contains a humectant. An emulsifier in a cream is usually a nonionic, anionic, cationic, or amphoteric surfactant. As with other carriers or excipients, the ointment base must be inert, stable, non-irritating and non-sensitizing.

In any of the above embodiments, the pharmaceutical compositions described herein may include one or more of one or more of the following components: lipids, interlayer cross-linked multilayer vesicles, biodegradable nanoparticles based on poly (D, L-lactic-co-glycolic acid ) [PLGA] or nanoparticles or microparticles based on polyanhydride and lipid bilayers based on nanoporous particles.

Dosages.

Dosages can vary depending on the requirements of the patient, the severity of the condition being treated, and the particular compound used. Determining the correct dosage for a particular situation can be determined by a medical professional. The total daily dose can be divided and administered in portions throughout the day or by continuous delivery.

In some embodiments, the compounds described herein are administered at a dosage of from about 0.001 mg / kg to about 500 mg / kg (e.g., from about 0.001 mg / kg to about 200 mg / kg; from about 0.01 mg / kg to about 200 mg / kg, from about 0.01 to about 150 mg / kg, from about 0.01 to about 100 mg / kg, from about 0.01 to about 50 mg / kg, from about 0.01 mg / kg to about 10 mg / kg, from about 0.01 to about 5 mg / kg, from about 0.01 to about 1 mg / kg, from about 0.01 to about 0.5 mg / kg, from about 0, 01 mg / kg to about 0.1 mg / kg, from about 0.1 mg / kg to about 200 mg / kg, from about 0.1 mg / kg to about 150 mg / kg, from about 0.1 mg / kg to about 100 mg / kg, from about 0.1 mg / kg to about 50 mg / kg, from about 0.1 mg / kg to about 10 mg / kg, from about 0.1 mg / kg to about 5 mg / kg, from about 0.1 mg / kg to about 1 mg / kg; from about 0.1 mg / kg to about 0.5 mg / kg).

Treatment regimens.

The above doses may be administered daily (e.g., as a single dose, or as two or more divided doses) or not daily (e.g. every other day, every two days, every three days, once a week, two weeks, once every two weeks, once a month).

In some embodiments, the administration period for the compound described herein is 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14 days, 3, 4, 5, 6 , 7, 8, 9, 10, 11, 12 weeks, 4, 5, 6, 7, 8, 9, 10, 11, 12 months or more.

In a further embodiment, the period during which the administration is stopped is 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14 days, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12 weeks, 4, 5, 6, 7, 8, 9, 10, 11, 12 months or more. In one embodiment, the therapeutic compound is administered to the individual over a period of time followed by a separate period of time. In another embodiment, the therapeutic compound is administered during a first period and a second period after the first period, the administration being stopped during the second period, followed by a third period when administration of the therapeutic compound begins, and then a fourth period after the third period when administration is stopped. In an aspect of this embodiment, the period of administration of the therapeutic compound followed by the period when the administration is stopped is repeated for a specified or indefinite period of time. In a further embodiment, the administration period is 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14 days, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12 weeks, 4, 5, 6, 7, 8, 9 10, 11, 12 months or more. In a further embodiment, the period during which the administration is stopped is 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14 days, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12 weeks, 4, 5, 6, 7, 8, 9, 10, 11, 12 months or more.

Treatment methods.

In some embodiments, methods are provided for treating a subject having a condition, disease, or disorder in which an increase in NLRP3 signaling can correct a deficiency in innate immune activity (eg, a condition, disease, or disorder associated with a deficient immune response) that contributes to the pathology, and / or symptoms, and / or the progression of a condition, disease, or disorder (eg, cancer).

Indications for use.

In any of the methods described herein, the subject may have cancer. In some examples of any of the methods described herein, the mammal has been identified as having cancer, or has been diagnosed as having cancer.

Non-limiting examples of cancer include acute myeloid leukemia, adrenocortical cancer, Kaposi's sarcoma, lymphoma, anal cancer, appendix cancer, teratoid / rhabdoid tumor, basal cell carcinoma, bile duct cancer, bladder cancer, bone cancer, brain cancer, breast cancer, bronchial tumor, carcinoid tumor, heart tumor, cervical cancer, chordoma, chronic lymphocytic leukemia, chronic myeloproliferative tumor, colon cancer, colorectal cancer, craniopharyngioma, bile duct cancer, endometrial cancer, ependymoma, esophageal cancer, esthesioneuroblastoma, cancer of the esophagus, esthesioneuroblastoma , fallopian tube cancer, gallbladder cancer, gastrointestinal carcinoid tumor, gastrointestinal stromal tumor, germ cell tumor, hair cell leukemia, head and neck cancer, heart cancer, liver cancer, hypopharyngeal cancer, pancreatic cancer, kidney cancer, laryngeal cancer, chronic myelogenous leukemia, lip and oral cavity cancer, lung cancer, glue melanoma exact Merkel carcinoma, mesothelioma, oral cancer, oral cancer, osteosarcoma, ovarian cancer, penile cancer, pharyngeal cancer, prostate cancer, cancer

- 70 037780 rectum, salivary gland cancer, skin cancer, small bowel cancer, soft tissue sarcoma, testicular cancer, throat cancer, thyroid cancer, urethral cancer, uterine cancer, vaginal cancer and vulvar cancer.

In certain embodiments, non-limiting examples of cancers include breast cancer, colon cancer, rectal cancer, rectal cancer, pancreatic cancer, and prostate cancer.

Methods for diagnosing a subject as having cancer or identifying a mammal as having cancer are well known in the art. For example, a medical professional (eg, a doctor, medical assistant, or technician) can diagnose cancer in a mammal by observing one or more of the symptoms of cancer in the mammal. Non-limiting examples of cancer symptoms include fatigue, a lump or area of thickening felt under the skin, change in mass, jaundice, darkening or redness of the skin, sores that do not heal, changes in existing moles, changes in bowel or bladder habits, persistent cough or problems with breathing, difficulty swallowing, hoarseness, persistent upset stomach or discomfort after eating, persistent, unexplained muscle or joint pain, persistent, unexplained fever or night sweats, and unexplained bleeding or bruising. Methods for diagnosing a subject as having cancer or identifying a subject as having cancer may further include performing one or more diagnostic tests (eg, performing one or more diagnostic tests on a biopsy or blood sample).

In some examples of any of the methods described herein, the subject may be a subject with cancer, a subject diagnosed with cancer, or a subject identified as having cancer who does not respond to previously prescribed cancer treatments. Diagnostic tests for diagnosing a subject as having cancer or identifying a mammal as having cancer are known in the art.

In some embodiments, methods are provided for treating a subject having a condition, disease, or disorder in which an increase in NLRP3 signaling can correct a deficiency in innate immune activity (e.g., a condition, disease, or disorder associated with a deficient immune response) that contributes to the pathology and / or symptoms and / or progression of a condition, disease, or disorder (eg, cancer).

In some embodiments, the present invention relates to a method for treating a cancer, wherein the cancer can be any cancer that does not elicit an optimal response from the innate immune system.

The innate immune system refers to the part of the immune system made up of cells that respond to threats to the body, such as infection or cancer, in an antigen-nonspecific manner and stimulate the adaptive antigen-specific immune system. In general, complete elimination of the threat and long-term protection (= immunity) requires the activity of the adaptive, antigen-specific immune system, which, in turn, depends on the stimulation of the innate immune system.

In some embodiments, the present invention relates to a method for treating a cancer selected based on resistance to T cell checkpoint inhibition, regardless of the type of cancer and based on failure to respond to previous T cell checkpoint inhibitor therapy or based on the type of cancer that typically resistant to therapy with T cell checkpoint inhibitors such as hormone receptor positive breast cancer, colon or rectal microsatellite cancer, pancreatic cancer, and prostate cancer.

In some other embodiments, the present invention provides a method for treating a malignant disease comprising an NLPR3 agonist of the present invention for treating noninflammatory tumors with low CD8 + T cell infiltration to enhance tumor immunogenicity and stimulate inflammatory responses. For example, the combination can be used to treat a solid tumor based on biopsy results that have demonstrated low CD8 + T cell infiltration or low expression of genes produced by CD8 + T cells.

Resistance to T-cell checkpoint inhibition refers to cancer progression during therapy or no response within 6 months of therapy according to consensus response criteria for the respective cancer, such as RECIST1.1 for most solid tumors.

T cell infiltration refers to the percentage of T cells of all nucleated cells as determined by immunohistochemistry of tumor biopsy samples.

CD8 + T cell infiltration refers to the percentage of CD8 + cells from all nucleated cells as determined by immunohistochemistry of tumor biopsy samples.

In addition to immunohistochemistry to quantify CD8 + T cells in biopsy samples, 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 reporting CD8 + infiltration T cells. Threshold values for low and high infiltration of CD8 + T cells when quantifying mRNA by immunohistochemistry are developed by various groups and take into account the spectrum of infiltration of CD8 + T-71 037780 cells in cancer, as well as in relation to specific types of cancer.

In any of the methods described herein, the subject may have an infectious disease. In some examples of any of the methods described herein, the subject has been identified as having an infectious disease, or has been diagnosed as having an infectious disease. For example, an infectious disease can be caused by a bacterium, virus, fungus, parasite, or mycobacterium.

Non-limiting examples of infectious disease include Acinobacter infection, actinomycosis, African sleeping sickness, acquired immunodeficiency syndrome, amebiasis, anaplasmosis, anthrax, Arcanobacterium haemolyticum infection, Argentine hemorrhagic fever, ascariasis, bacteriosis, aspergillosis, babies ceremonia , bacterial vaginosis, Bacteroides infection, balantidiasis, Baylisascaris infection, BK viral infection, black piedra, Blastocystic hominis infection, blastomycosis, Bolivian haemorrhagic fever, botulism, Brazilian haemorrhagic fever, Burkurulosis, Burkurulosa infection, brucellosis Calicivirus, camptobacteriosis, candidiasis, cat scratch disease, cellulitis, Chagas disease, chancroid, chickenpox, chikungunya, chlamydia, Chlamydophila pneumoniae infection, cholera, chromoblastomycosis, clonorchiasis, Clostridium difficile infection, coccidiomycosis Orada tick-borne fever, viral infection of the upper respiratory tract, Creutzfeldt-Jakob disease, Crimean-Congo hemorrhagic fever, crytococcosis, cryptosporidiosis, migratory skin larva, Cyclosporosis, cysticercosis cysticercosis, infection , diphyllobothriasis, dracunculiasis, hemorrhagic fever Ebola, echinococcosis, ehrlichiosis, enterobiasis, Enterococcus infection, Enterovirus infection, epidemic typhus, erythema infection, exanthema, fascioloptiosis, fasciolosis, fatal familial insomnia amoebae, Fusobacterium infection, gas gangrene, geotrichosis, Gerstmann-Straussler-Scheinker syndrome, giardiosis, glanders, gnatostomosis, gonorrhea, inguinal granuloma, group A streptococcal infection, group A streptococcal infection, group B streptococcal influenza Russian vesicular stomatitis, hantavirus pulmonary syndrome, Heartland virus disease, Heliobacter pylori infection, hemolytic uremic syndrome, hemorrhagic fever with renal syndrome, hepatitis A, hepatitis B, hepatitis C, hepatitis D, hepatitis E, herpes simplex, histoplasmosis , human bocavirus infection, human ehrlichiosis, anaplasmosis of human granulocytes, human metaplevomyovirus infection, human monocytic ehrlichiosis, human papillomavirus infection, human parainfluenza virus infection, hymenolepase caused by the Epstein-Barr virus (Epstein-Barr disease), infectious disease , keratitis, Kingella kingae infection, kuru, Lassa fever, Legionnaires' disease, Pontiac fever, leishmaniasis, leprosy, leptospirosis, listeriosis, Lyme disease, lymphatic filariasis, lymphocytic choreomeningitis, malaria, Marburg hemorrhagic fever, mesenteric fever syndrome t, meningococcal infection, metagonimiasis, microsporidiosis, molluscum contagiosum, monkeypox, mumps, rat typhus, mycoplasma pneumonia, mycetoma, myiasis, neonatal conjunctivitis, a variant of Creutzfeldt-Jakob disease, paradiocytosis, paradiocytosis, onchocardiosis , head lice, pubic lice, pelvic inflammation, pertussis, plague, pneumonia, poliomyelitis, Prevotella infection, primary amoebic meningoencephalopathy, progressive multifocal leukoencephalopathy, psittacosis, Q fever, rabies, recurrent rheumatoid arthritis , rickettsial infection, rickettsiosis, Rift Valley fever, Rocky Mountain spotted fever, rotavirus infection, rubella, salmonellosis, severe acute respiratory syndrome, scabies, schistosomiasis, sepsis, shigellosis, herpes zoster, staphylitis ovarian infection, staphylococcal infection, strongyloidosis, subacute sclerosing panencephalitis, syphilis, teniasis, tetanus, ringworm barabe, ringworm, fungal infection of the smooth skin of the trunk, inguinal dermatomycosis, fungal infection of the skin of the hands, black fungus, feet pityriasis versicolor, toxocariasis, trachoma, toxoplasmosis, trichinosis, trichomoniasis, trichuriasis, tuberculosis, tularemia, typhoid fever, Ureaplasma urealyticum infection, valley fever, Venezuelan hemorrhagic fever, pyocytic pneumoniae, Western fever, Nile fever yersineosis, yellow fever and zygomycosis.

Methods for diagnosing a subject as having an infectious disease or identifying a subject as having an infectious disease are well known in the art. For example, a medical professional (eg, a physician, physician assistant, or technician) can 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 a mammal as having an infectious disease or identifying a subject as having an infectious disease may further comprise performing one or more diagnostic tests (eg, performing one or more diagnostic tests on a biopsy or blood sample). Diagnostic tests for diagnosing a subject as having an infectious disease or identifying a subject as having an infectious disease are known in the art.

Combined therapy.

This disclosure addresses both monotherapy and combination therapy regimens.

In some embodiments, the methods described herein may further comprise administering one or more complementary therapies (e.g., one or more complementary therapeutic agents and / or one or more treatment regimens) in combination with the administration of the compounds described herein.

In certain embodiments, the methods described herein may further comprise administering one or more additional anti-cancer therapies.

One or more complementary cancer therapies may include, but are not limited to, surgery, radiation therapy, chemotherapy, toxotherapy, immunotherapy, cryotherapy, cancer vaccines (eg, HPV vaccine, hepatitis B vaccine, Oncophage, Provenge), gene therapy, and their combinations. Immunotherapy, including, but not limited to, adoptive cell therapy, stem cell and / or dendritic cell production, blood transfusion, lavage, and / or other treatments including, but not limited to, tumor freezing.

In some embodiments, the one or more additional anti-cancer therapies is chemotherapy, which may include the administration of one or more additional chemotherapeutic agents.

In certain embodiments, the additional anti-cancer therapy comprises (chemotherapeutic agent) an immunomodulatory moiety, eg, an immune checkpoint inhibitor. In some of these embodiments, an immune checkpoint inhibitor targets an immune checkpoint receptor selected from CTLA-4, PD-1, PD-L1, PD-1-PDL1, PD-1-PD-L2, T cell immunoglobulin, and mucin 3 (TIM3 or HAVCR2), Galectin 9 - TIM3, phosphatidylserine - TIM3, lymphocyte activation gene protein 3 (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, HVEMCD160, HVEM-LIGHT, HVEM-BTLA-CD160, CD80, CD80 - PDL-1, PDL2 - CD80, CD244, CD48 - CD244, CD244, ICOS, ICOS-ICOS ligand, B7-H3, B7-H4, VISTA, TMIGD2, HHLA2-TMIGD2, butyrophilins, including BTNL2, Siglec family, members of the TIGIT family and PVR, 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, neuropilin, CD160 , CD30 and CD155 (e.g. CTLA-4 or PD1 or PD-L1) and other immunomodulatory agents such as interleukin-2 (IL-2), indoleamine2,3-dioxygenase (IDO), IL-10, transforming growth factor-β (TGFP), CD39, CD73 Adenosine CD39-CD73 and CXCR4-CXCL12. See, for example, Postow M.J. Clin. Oncol. 33, 1 (2015).

In certain embodiments, an 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.

In certain embodiments, the immune checkpoint inhibitor is selected from nivolumab (also known as OPDIVO; previously designated 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), JS001 (TAIZHOU JUNSHI PHARMA; see 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; see WO 2015/085847; Si-Yang Liu et al., J. Hematol. Oncol. 10: 136 (2017)), TSR-042 (ANB011; Tesaro Biopharmaceutical ; see WO 2014/179664), GLS-010 (WBP3055; Wuxi / Harbin Gloria Pharmaceuticals; see Si-Yang Liu et al., J. Hematol. Oncol. 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, WO 2017/133540); BMS-936559 (formerly 12A4 or MDX1105; see, for example, US Patent No. 7943743 and WO 2013/173223), MPDL3280A (also known as RG7446, atezolizumab and TECENTRIQ; US 8217149; see also Herbst et al. (2013) J Clin Oncol 31 (Suppl): 3000), durvalmab (IMFINZI; MEDI-4736; AstraZeneca; see WO 2011/066389), avelumab (Pfizer; MSB0010718C; BAVENCIO; see WO 2013/079174)), STI- 1014 (Sorrento; see WO 2013/181634), CX-072 (Cytomx; see WO 2016/149201), KN035 (3D Med / Alphamab; see Zhang et al., Cell Discov. 7: 3 (March 2017) , LY3300054 (Eli Lilly Co .; see, for example, WO 2017/034916), CK-301 (Checkpoint Therapeutics; see Gorelik et al., AACR: Abstract 4606 (Apr 2016)); urelumab, PF-05082566, MEDI6469 , TRX518, varlilumab, CP870893, BMS-986016, MGA271, lirilumab, IPH2201, emactuzumab, INCB024360, galunizertiba, ulocuplumaba, BKT140, Bavituximab, Ss-90002, MNERVILIP;

- 73 037780 US patent No. 698420), MK-1308 (Merck), AGEN-1884 (Agenus Inc .; WO 2016/196237) and tremelimumab (formerly ticilimumab, CP-675.206; AstraZeneca; see, for example, WO 2000/037504 and Ribas, Update Cancer

Ther. 2 (3): 133-39 (2007)).

In certain embodiments, the immune checkpoint inhibitor is selected from: nivolumab, pembrolizumab, JS001, BGB-A317, INCSHR1210, TSR-042, GLS-010, STI-1110, MGD013, IBI308, BMS-936559, atezolizumab, durvalumab, STI avelumab -1014, CX-072, KN035, LY3300054, CK-301, urelumab, PF-05082566, MEDI6469, TRX518, varlilumab, BMS-986016, ipilimumab, AGEN1884, and tremelimumab.

In some of these embodiments, the immune checkpoint inhibitor is selected from urelumab, PF-5082566, MEDI6469, TRX518, varlilumab, CP-870893, pembrolizumab (PD1), nivolumab (PD1), atezolizumab (formerly MPDL3280A), PDI361 (PDL1 -L1), avelumab (PD-L1), PDR001 (PD1), BMS-986016, mGa271, lyrilumab, IPH2201, emactuzumab, INCB024360, galunisertiba, ulocuplumab, BKT140, Bavituximab, CCN-90002P85A

In certain embodiments, the immune checkpoint inhibitor is selected from nivolumab, ipilimumab, pembrolizumab, atezolizumab, durvalumab, and avelumab.

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

In certain embodiments, the additional anti-cancer agent (chemotherapeutic agent) is a STING agonist. For example, a STING agonist may include cyclic dinucleotides such as cAMP, cGMP, and cG AMP, and modified cyclic dinucleotides that include one or more of the following modification traits (2'O / 3'-O bond, phosphorothioate bond, adenine and / or a guanine analogue modification 2'-OH (eg —OCH3 or substitution eg —F or N ₃ ) See, eg, WO 2014/189805 In certain embodiments, the additional chemotherapeutic agent is an alkylating agent. so named because of their ability to alkylate many nucleophilic functional groups under conditions present in cells, including but not limited to cancer cells.In an additional embodiment, an alkylating agent includes, but is not limited to, cisplatin, carboplatin, mechlorethamine, cyclophosphamide, chlorambucil, ifosfamide, and / or oxaliplatin In one embodiment, alkylating agents can function to disrupt cellular function by forming covalent bonds with amino, carboxyl, sulfhydryl and phosphate groups in biologically important molecules, or they can work by modifying cell DNA. In a further embodiment, the alkylating agent is synthetic, semi-synthetic, or derived.

In certain embodiments, the additional chemotherapeutic agent is an antimetabolite. Antimetabolites disguise themselves as purines or pyrimidines, the building blocks of DNA and, in general, prevent the inclusion of these substances in DNA during the S phase (cell cycle), stopping normal development and division. Antimetabolites can also interfere with RNA synthesis. In one embodiment, the antimetabolite includes, but is not limited to, azathioprine and / or mercaptopurine. In a further embodiment, the antimetabolite is synthetic, semi-synthetic, or derived.

In certain embodiments, the additional chemotherapeutic agent is a plant alkaloid and / or terpenoid. These alkaloids come from plants and block cell division, generally preventing microtubule function. In one embodiment, the plant alkaloid and / or terpenoid is vinca alkaloid, podophyllotoxin and / or taxane. Vinca alkaloids tend to bind to specific regions of tubulin, inhibiting the assembly of tubulin into microtubules, usually during the M phase of the cell cycle. In one embodiment, the vinca alkaloid is, without limitation, derived from the Madagascar vinca, Catharanthus roseus (formerly known as Vinca rosea). In one embodiment, the vinca alkaloid includes, but is not limited to, vincristine, vinblastine, vinorelbine, and / or vindesine. In one embodiment, the taxane includes, but is not limited to, taxol, paclitaxel, and / or docetaxel. In a further embodiment, the plant alkaloid or terpernoid is synthetic, semi-synthetic, or derived. In an additional embodiment, the podophyllotoxin is, without limitation, etoposide and / or teniposide. In one embodiment, the taxane is, without limitation, docetaxel and / or ortataxel. In one embodiment, the anti-cancer therapeutic agent is a topoisomerase. Topoisomerases are important enzymes that maintain DNA topology. Inhibition of type I or type II topoisomerases interferes with both transcription and DNA replication, disrupting proper DNA supercoiling. In an additional embodiment, the topoisomerase is, without limitation, a type I topoisomerase inhibitor or a type II topoisomerase inhibitor. In one embodiment, the type I topoisomerase inhibitor is, without limitation, camptothecin. In another embodiment, camptothecin is, without limitation, exeatecan, irinotecan, lurtotecan, topotecan, BNP 1350, CKD 602, DB 67 (AR67) and / or ST 1481. In one embodiment, the type II topoisomerase inhibitor is , without limitation, epipodophyllotoxin. In an additional embodiment, the epipodophyllotoxin is, without limitation, amsacrine, etoposide, etoposide phosphate, and / or teniposide. In a further embodiment, the topoisomerase is synthetic, semi-synthetic or derivative, including those that occur naturally, such as, without limitation, epipodophyllotoxins, substances naturally occurring in the root of the American Mayapple (Podophyllum peltatum).

In certain embodiments, the additional chemotherapeutic agent is a stilbenoid. In an additional embodiment, the stilbenoid includes, but is not limited to, Resveratrol, Piceatannol, Pinosylvin, Pterostilbene, Alpha Viniferin, Ampelopsin A (Ampelopsin A) (Ampelopsin A) (Ampelopsin E), Diptoindonesin C (Diptoindonesin C), Diptoindonin F (Diptoindonesin F), Epsilon-Vinferin (Epsilon-Vinferin), Flexuosol A (Flexuosol A), Gnetin N (Gnetin N), Gemsleanol D (Hemsleyanol D), Hemsleyo (Hopeaphenol), Trans-Diptoindonesin B (Trans-Diptoindonesin B), Astringin (Astringin), Piceid and Diptoindonesin A (Diptoindonesin A). In a further embodiment, the stilbenoid is synthetic, semi-synthetic, or derived.

In certain embodiments, the additional chemotherapeutic agent is a cytotoxic antibiotic. In one embodiment, the cytotoxic antibiotic is, without limitation, actinomycin, anthracenedione, anthracycline, thalidomide, dichloroacetic acid, nicotinic acid, 2-deoxyglucose, and / or chlofazimine. In one embodiment, actinomycin is, without limitation, actinomycin D, bacitracin, colistin (polymyxin E), and / or polymyxin B. In another embodiment, anthracenedione is, without limitation, mitoxantrone and / or pixantrone. In an additional embodiment, the anthracycline is, without limitation, bleomycin, doxorubicin (adriamycin), daunorubicin (daunomycin), epirubicin, idarubicin, mitomycin, plikamycin and / or valrubicin. In an additional embodiment, the cytotoxic antibiotic is synthetic, semi-synthetic, or derived.

In certain embodiments, the additional chemotherapeutic agent is selected from endostatin, angiogenin, angiostatin, chemokines, angioarrestin, angiostatin (plasminogen fragment), anti-angiogenic factors derived from basement membrane collagen (tumstatin, canstatin, or arrestin), transangiogenic antithrombin III inhibitor cartilage origin (CDI), CD59 complement fragment, fibronectin fragment, gro-β, heparinases, heparin hexasaccharide fragment, human chorionic gonadotropin (hCG), α / β / γ interferon, interferon-induced protein (IP-10), interleukin-12 , kringle 5 (plasminogen fragment), metalloproteinase (TIMP) inhibitors, 2-methoxyestradiol, placental ribonuclease inhibitor, plasminogen activator inhibitor, platelet factor-4 (PF4), 16 kDa prolactin fragment, proliferin-related protein (PRP), various retinoids tetrahydrocortisol-S, thrombospondin-1 (TSP-1), transforming fa growth factor-beta (TGF-β), vasculostatin, vasostatin (calreticulin fragment)) and the like.

In certain embodiments, the additional chemotherapeutic agent is selected from abiraterone acetate, altretamine, anhydrovinblastine, auristatin, bexarotene, bicalutamide, BMS 184476, 2,3,4,5,6-pentafluoro-N- (3-fluoro-4-methoxyphenyl) benzenesulfonamide, bleomycin, N, Ndimethyl-L-valyl-L-valyl-N-methyl-L-valyl-L-proli-1-L-proline-tert-butylamide, cachectin, cemadotin, chlorambucil, cyclophosphamide, 3 ', 4'- didehydro-4'-deoxy-8'-norvin-kaleikoblastine, docetaxol, docetaxel, cyclophosphamide, carboplatin, carmustine, cisplatin, cryptophycin, cyclophosphamide, cytarabine, dacarbazine-decibuxin (ditic), cytacabucinarabine) (diticabucinacin , decibuxin, dacticubin (molecule)), etoposide, 5-fluorouracil, finasteride, flutamide, hydroxyurea, and hydroxyureataxanes, ifosfamide, liarosol, lonidamine, lomustine (CCNU), MDV3100, mechlorethamine, mechlorethene, mechlorethamine (mechlorethine) , streptozocin, mitomycin, methotrexate, taxanes, nilutamide, onapristone, paclitaxel, prednimustine, procarbazine, RPR109881, stramustine phosphate, tamoxifen, tazonermine, taxol, tretinoin, vinblastine, vincristine, vindesine sulfate and vinflunine.

In certain embodiments, the additional chemotherapeutic agent is platinum, cisplatin, carboplatin, oxaliplatin, mechlorethamine, cyclophosphamide, chlorambucil, azathioprine, mercaptopurine, vincristine, vinblastine, vinorelbine, vindesine, etoposide and tenipelinacetane, paclitoside, paclitoside , etoposide phosphate, teniposide, 5-fluorouracil, leucovorin, methotrexate, gemcitabine, taxane, leucovorin, mitomycin C, tegafururacil, 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 deforolimus.

In still other embodiments, the additional chemotherapeutic agent may be selected from those described in US Pat. No. 7,927,613.

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In yet another embodiment, the methods may further comprise administering one or both of: (i) one or more antifungal agents (e.g., selected from the group consisting of bifonazole, butoconazole, clotrimazole, econazole, ketoconazole, luliconazole, miconazole, omoconazole, oxyconazole, sertaconazole , sulconazole, thioconazole, albaconazole, efinaconazole, eposiconazole, fluconazole, isavuconazole, itraconazole, posaconazole, propiconazole, ravuconazole, terconazole, voriconazole, abafungin, amorolfine, butenafinafinung, caabafinafluxin -fluorocytosine, griseofulvin, haloprogyn, tolnaflate, undecylenic acid and balm of Peru) and (ii) one or more antibiotics (for example, selected from the group of amikacin, gentamicin, kanamycin, neomycin, netilmicin, tobramycin, paromycin, spectomycin , rifaximin, loracarbef, ertapenem, doripenem, imipenem, cyl astatine, meropenem, cefadroxil, cefazolin, cephalothin, cephalothin, cephalexin, cefaclor, tsefamandola, cefoxitin, cefprozil, cefuroxime, cefixime, cefdinir, tsefditorena, cefoperazone, cefotaxime, cefpodoxime, ceftazidime, ceftibuten, tseftizoksima, ceftriaxone, cefepime, ceftaroline fozamila, ceftobiprole , teicoplanin, vancomycin, telavancin, dalbavantsina, oritavancin, clindamycin, lincomycin, daptomycin, azithromycin, clarithromycin, dirithromycin, erythromycin, roxithromycin, troleandomycin, telithromycin, spiramycin, aztreonam, furazolidone, nitrofurantoin, linezolid, posizolida, radezolida, torezolida, amoxicillin, ampicillin , azlocillin, carbenicillin, cloxacillin, dicloxacillin, flucloxacillin, mezlocillin, methicillin, nafcillin, oxacillin, penicillin G, penicillin V, piperacillin, penicillin G, temotsillina, ticarcillin, amoxicillin, kalvulanata, ampicillin, ampicillin bibaktiltsillotsillotsila, tsibaktsillina, ambiltsiltsina, Cipro floksatsina, enoxacin, gatifloxacin, gemifloxacin, levofloxacin, lomefloksa CIN, moxifloxacin, nalidixic acid, norfloxacin, ofloxacin, trovafloxacin grepafloksatsina, sparfloxacin, temafloxacin, mafenida, sulfacetamide, sulfadiazine, silver sulfadiazine, sulfadimethoxine, sulfamethoxazole, sulfanilamide, sulfasalazine, sulfisoxazole, trimetoprimsulfametoksazola , sulfonamideochrysoidine, demeclocycline, minocycline, oxytetracycline, tetracycline, clofazimine, dapsone, dapreomycin, cycloserine, ethambutol, ethionamide, isoniazid, pyrazinamide, rifampicin, rifabutin, rifapentine, fosfapentine, streptomycinoma acid quinupristine, dalopristin, thiamphenicol, tigecycyline, tinidazole, trimethoprim, and teixobactin).

In certain embodiments, the second therapeutic agent or dosing regimen is administered to the subject prior to contact with the chemical or administration of the chemical (e.g., about an 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).

In other embodiments, the second therapeutic agent or dosing regimen is administered to the subject at about the same time as when the chemical is contacted or administered. By way of example, a second therapeutic agent or regimen and a chemical are administered to a subject simultaneously in the same dosage form. As another example, the second therapeutic agent or regimen and the chemical are administered to a subject simultaneously in separate dosage forms.

In yet other embodiments, the second therapeutic agent or dosing regimen is administered to the subject following contact with or administration of the chemical (e.g., about one hour after, or about 6 hours after, or about 12 hours after, or about 24 hours after , or about 48 hours, or about 1 week after, or about 1 month after).

Patient selection.

In some embodiments, the methods described herein further include the step of identifying a subject (eg, a patient) in need of such treatment (eg, by biopsy, endoscopy, or other conventional method known in the art). In certain embodiments, the NLRP3 protein can serve as a biomarker for certain types of cancer.

In some embodiments, the chemicals, methods, and compositions described herein can be administered to specific treatment-resistant patient populations (eg, patients resistant to checkpoint inhibitors).

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

In some embodiments, a compound of the present invention, or a pharmaceutically acceptable salt thereof, or a pharmaceutical composition containing it, can be used as a medicament. In certain embodiments, the compounds of the present invention can be used for the manufacture of a medicament for the treatment of cancer.

Examples of

As one skilled in the art will appreciate, methods for synthesizing compounds of the formulas provided herein will be apparent to those skilled in the art. For example, the compounds described herein can be synthesized, for example, using one or more of the methods described herein and / or using the methods described, for example, in US 2015/0056224. Synthetic chemistry transformations and protecting group (protection and deprotection) techniques used in the synthesis of the compounds described herein are known in the art and include, for example, those described in R. Larock, Comprehensive Organic Transformations. VCH Publishers (1989); T.W. Greene and RGM. Wuts, Protective Groups in Organic Synthesis, 2d. Ed., John Wiley and Sons (1991); 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. The starting materials used in the preparation of the compounds of the invention are known, prepared by known methods, or are commercially available. One skilled in the art will also understand that the conditions and reagents described herein may be replaced by alternative equivalents recognized in the art. For example, in many reactions, triethylamine can be interchangeable with other bases such as non-nucleophilic bases (eg, diisopropylamine, 1,8-diazabicycloundec-7-ene, 2,6-di-tert-butylpyridine, or tetrabutylphosphazene).

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

To further illustrate the foregoing, the following non-limiting exemplary synthesis schemes are included. Variations of these examples within the scope of the claims are within the skill of the art and are considered to be within the scope of the invention as described and claimed herein. The reader will understand that a person skilled in the art provided by the present disclosure and a person skilled in the art can prepare and apply the invention without exhaustive examples.

The following abbreviations have the meanings mentioned:

ACN - acetonitrile;

CH ₂ Cl ₂ - dichloromethane;

CH ₃ ReO ₃ - methyltrioxorhenium;

Cs ₂ CO ₃ - cesium carbonate;

d - doublet;

DCM for dichloromethane;

DIEA - N, N-diethylisopropylamine;

DMF - N, N-dimethylformamide;

DMSO - dimethyl sulfoxide;

ES - electrospray ionization;

EA, EtOAc - ethyl acetate;

EtOH is ethanol;

eq - equivalents;

g - gram;

h - hours;

HCl - hydrogen chloride (usually in the form of a solution);

H2O - water;

H ₂ O ₂ - hydrogen peroxide;

HATU - 1- [bis (dimethylamino) methylene] -1H-1,2,3-triazole [4,5-b] pyriginium 3-oxide hexafluorophosphate;

HPLC - high performance liquid chromatography;

K2CO3 - potassium carbonate;

LC / MS - liquid chromatographic mass spectrometer;

LiBH ₄ - lithium borohydride;

m - multiplet;

M - molar;

m-CPBA - metachloroperoxybenzoic acid;

- 77,037,780 mg - milligram;

MeOH - methanol;

MHz - megahertz;

ml - milliliter (s);

mmol - millimole (s);

NaHCO ₃ - sodium bicarbonate;

Na2CO ₃ - sodium carbonate;

NaOH - sodium hydroxide;

Na2CO ₃ - sodium sulfate;

NEt ₃ - trimethylamine;

NH3 is ammonia;

NH ₄ OH or NH3H2O - ammonium hydroxide;

NH ₄ HCO ₃ - ammonium hydrogen carbonate;

nm - nanometer;

PdCl2 (PPh ₃ ) 2 - bis (triphenylphosphine) palladium (II) dichloride;

Pd (dppf) Cl ₂ - [1,1'-bis (diphenylphosphino) ferrocene] dichloropalladium (II);

PE - petroleum ether;

PMB - para-methoxybenzyl;

POCl ₃ - phosphorus oxychloride;

ppm - parts per million;

Ru - pyridine;

s - singlet;

t - triplet;

T3P - 2,4,6-tripropyl-1,3,5,2,4,6-trioxatriphosphorinan-2,4,6-trioxide;

TFA, trifluoroacetic acid;

TLC - thin layer chromatography;

TsCl = para-toluenesulfonyl chloride chloride;

° С - degrees Celsius;

micromolar - micromolar.

General procedure for the synthesis of compounds of formula I.

The compounds of the present invention can be prepared by a number of methods well known to those skilled in the art of organic synthesis. The compounds of the present invention can be synthesized using the methods described below, together with synthetic methods known in the art of synthetic organic chemistry, or variations thereof that are appreciated by those skilled in the art. Preferred methods include, but are not limited to, the methods described below.

Compounds of this invention can be prepared using the reactions and procedures described in this section. The reactions are carried out in solvents appropriate to the reagents and materials used and suitable for the transformations to be carried out. In addition, when describing the synthesis methods described below, it should be understood that all proposed reaction conditions, including the choice of solvent, the reaction atmosphere, the reaction temperature, the duration of the experiment and the processing procedures, are chosen as standard conditions for this reaction, which should be easily understood. a person skilled in the art. One of ordinary skill in the art of organic synthesis understands that functional groups present at various sites on the molecule must be compatible with the reagents and proposed reactions. Such restrictions on substituents that are compatible with the reaction conditions will be apparent to a person skilled in the art, and then alternative methods must be used. Sometimes it will be necessary to change the order of the synthesis steps or to prefer one particular process scheme over another in order to obtain the desired compound of the invention. It is also recognized that another important factor in planning any synthetic route in the art is the judicious choice of the protecting group used to protect the reactive functional groups present in the compounds described herein. An authoritative report describing many alternative paths to the trained practitioner is Greene and Wuts (Protective Groups In Organic Synthesis, Third Edition, Wiley and Sons, 1999).

Compounds of formula (I) can be prepared by reference to the methods illustrated in the following schemes. As shown herein, the final product is a compound having the structural formula of formula (I). It is understood that any compound of formula (I) can be prepared according to the aforementioned schemes by a suitable selection of reagents with suitable substitution. Solvents, temperatures, pressures, and other reaction conditions can be readily selected by one of ordinary skill in the art. The starting materials are commercially available or can be readily obtained by a person skilled in the art. Components of the compounds are as defined herein or elsewhere in the description.

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Synthesis of compounds of formula (I) can be carried out using the methods summarized in Scheme 1.

Scheme 1

Stage 1. The first stage of Scheme 1 begins with a suitably functionalized quinolinol (i). If desired, the R ⁸ and R ⁹ groups can be the R ⁴ and R ⁵ groups found in the final product (xii). Alternatively, one or both of R ⁸ and R ⁹ can be groups that can be modified at a later stage of the synthesis, such as bromine. The first step of Scheme 1 can be performed by treating compound (i) with a suitable nitrating agent such as nitric acid in a suitable solvent such as propionic acid at a suitable temperature such as 130 ° C. to provide compound (ii).

Step 2. The second step of Scheme 1 can be performed by treating compound (ii) with a suitable chlorinating agent, such as phosphorus oxychloride, at a suitable temperature, such as 120 ° C., to provide compound (iii).

Step 3. In the third step of Scheme 1, the established R ¹⁰ group may be the desired R ³ group found in the final product (xii). Alternatively, a protecting group such as pmethoxybenzyl can be set and removed at a later stage in the synthesis. Step 3 can be carried out by treating compound (iii) with a suitable amine such as p-methoxybenzylamine and a base such as triethylamine in a solvent such as dichloromethane at a suitable temperature such as room temperature to give compound (iv) ...

Step 4. The fourth step of Scheme 1 can be performed by treating compound (iv) with a suitable reducing agent such as stannous chloride (II) in a solvent such as ethanol at a suitable temperature such as 65 ° C. to provide compound (v) ...

Step 5. In the fifth step of Scheme 1, the specified R ¹¹ and R ¹² groups may be the R ¹ and R ² groups required in the final product (xii). Alternatively, they can be groups that allow R ¹ and / or R ² to be set at a later stage, such as the Boc protecting group. Stage 5 can be carried out by treating compound (V) with an appropriate

79 037780 with a functionalized carboxylic acid such as N-Boc-N-ethylglycine, a suitable coupling agent such as HATU, and a suitable amine such as triethylamine, in a solvent such as dichloromethane at an appropriate temperature such as room temperature, to give compound (vi).

Step 6. The sixth step of Scheme 1 can be performed by treating compound (vi) with a suitable base such as triethylamine in a solvent such as ethanol at a suitable temperature such as 70 ° C. to provide compound (vii).

Step 7. The seventh step of Scheme 1 can be performed by treating compound (vii) with a suitable oxidizing agent such as m-CPBA in a solvent such as dichloromethane to provide compound (viii).

Step 8. The eighth step of Scheme 1 can be performed by treating compound (viii) with a reagent such as tosyl chloride and an amine such as ammonia in a solvent such as dichloromethane to give compound (ix).

Steps 9, 10 and 11 of Scheme 1 can be carried out to convert one or more of the groups R ⁸ , R ⁹ , R ¹⁰ , R ¹¹ and R ¹² to the groups R ¹ , R ² , R ³ , R ⁴ and R ⁵ required in final product (xii). One of ordinary skill in the art will understand that one or more of these steps may be optional depending on the reagents selected earlier in the synthesis and the desired end product. In addition, one skilled in the art will understand that these steps can be performed in an alternative order depending on the desired end product.

Step 9. The ninth step of Scheme 1 can optionally be carried out to convert one or both of the R ⁸ and R ⁹ groups to the R ⁴ and R ⁵ groups required in the final product. For example, if R ⁹ is bromo and the desired R ⁵ is 3-pyrazoyl, this transformation can be achieved by treating compound (ix) with a suitable boronic ester such as 3 (tetramethyl-1,3,2-dioxaborolane) -2 -yl) -7H-pyrazole in the presence of a catalyst such as a complex of dichloromethane Pd (dppf) Cl ₂ and a base such as cesium carbonate in a solvent mixture such as dioxane / water at a suitable temperature such as 100 ° C, to get connection (x). Alternatively, if R ⁹ is bromine and the desired R ⁵ is 1-pyrazolyl, this step can be carried out by treating compound (ix) with pyrazole in the presence of a catalyst such as copper (I) iodide, a ligand such as N, N'-dimethyl-ethylenediamine, and a base such as sodium carbonate in a solvent such as dimethyl sulfoxide at an appropriate temperature, such as 120 ° C., to give compound (x).

Step 10. The tenth step of Scheme 1 can optionally be carried out to convert one or both of the R ¹¹ and R ¹² groups to the R ¹ and R ² groups required in the final product. If one or both of the groups R ¹¹ and R ¹² represent a protecting group such as Boc, this group can be removed under suitable conditions, such as treatment of compound (x) with HCl in dioxane. If R ¹¹ and / or R ¹² is H and the desired R ¹ and / or R ² is an amide, this step can be carried out by treating compound (x) with a suitable anhydride and a base such as acetic anhydride and triethylamine in a solvent such as dichloromethane, or by treating compound (x) with a suitable acid, a coupling agent and a base such as 3-hydroxy-3-methylbutanoic acid, 2,4,6-tripropyl-1,3,5,2,4,6 β-trioxatriphosphinan, 2,4,6-trioxide, and Hunig's base, in a solvent such as DMF, to give compound (xi). Alternatively, if R ¹¹ and / or R ¹² is H and the desired R ¹ and / or R ² is an amine, this step can be carried out by treating compound (x) with a suitable aldehyde or ketone such as isobutyraldehyde, and a reducing agent such as sodium triacetoxyborohydride in a solvent such as methanol to give compound (xi). Alternatively, if R ¹¹ and / or R ¹² is H and the desired R ¹ and / or R ² is a carbamate, this step can be carried out by treating compound (x) with a suitable chloroformate such as ethyl chloroformate and a suitable a base such as triethylamine in a solvent such as dichloromethane, followed by treatment with a suitable base such as triethylamine in an alcoholic solvent such as methanol to give compound (xi). Alternatively, if R ¹¹ and / or R ¹² is H and the desired R ¹ and / or R ² is urea, this step can be carried out by treating compound (x) with a suitable carbamoyl chloride such as morpholine-4-carbonyl chloride , or an appropriate isocyanate such as ethyl isocyanate in the presence of a base such as Hunig's base in a solvent such as DMF. Alternatively, if R ¹¹ and / or R ¹² is H and the desired R ¹ and / or R ² is a sulfonylurea, this step can be carried out by treating compound (x) with a suitable sulfamoyl chloride such as dimethylsulfamoyl chloride and a base, such as Hunig's base in a solvent such as DMF to give compound (xi). Alternatively, if R ¹¹ and / or R ¹² is H and the desired R ¹ and / or R ² is a sulfonamide, this step can be carried out by treating compound (x) with a suitable sulfonyl chloride and a base such as 2-propylsulfonyl chloride and Junig's base in a solvent such as DMF.

Stage 11. The eleventh stage of Scheme 1 can optionally be carried out to convert

80 037780 of the R ¹⁰ group to the R ³ group desired in the final product (xii). For example, if R ¹⁰ is a protecting group such as p-methoxybenzyl, it can be removed under appropriate conditions, for example, treatment with trifluoroacetic acid at a suitable temperature, such as 70 ° C, to provide compound (xii).

Example 1. Procedure for the synthesis of 7-bromo-N ⁴ - (4-methoxybenzyl) quinoline-3,4-diamine.

Scheme 2

7-Bromoquinolin-4-ol (4.5 g, 20.0 mol) was dissolved in propionic acid (34 ml). The mixture was heated to 130 ° C and nitric acid (1.7 ml, 70%) was added. The reaction mixture was heated at 130 ° C (bath temperature) for 4 hours, after which it was cooled to room temperature and filtered. The resulting solid was washed with water (3x20 ml), 2-propanol (20 ml) and hexane (20 ml). The product was then dried under high vacuum to give 3.8 g (70.6%) of 7-bromo-3-nitroquinoline-4-ol as a tan powder, which was used in the next step without further purification. (ES, m / z): [M + H] + = 269.2 / 271.3.

Stage 2. Obtaining 7-bromo-4-chloro-3-nitroquinoline.

7-Bromo-3-nitroquinolin-4-ol (3.8 g, 14.12 mmol) was suspended in POCl ₃ (20 ml). Anhydrous DMF (1 ml) was added. The mixture was then heated to 120 ° C under nitrogen atmosphere for 3 h, after which the reaction mixture was cooled to room temperature. The precipitate was collected by filtration, washed with water and then partitioned between CH ₂ Cl ₂ (60 ml) and saturated aqueous Na ₂ CO ₃ . The organic layer was separated, washed with brine, dried over Na ₂ SO ₄ , filtered and concentrated to give 3.3 g (11.5 mmol, 81%) of 7-bromo-4-chloro-3-nitroquinoline as a beige solid colors. (ES, m / z): [M + H] + = 287.1 / 288.9.

Note: It has been found that when higher temperatures and longer reaction times are used, a significant amount of Cl-Br exchange occurs, resulting in an intermediate that does not undergo subsequent cross-coupling reactions.

Stage 3. Obtaining 7-bromo-N- (4-methoxybenzyl) -3-nitroquinolin-4-amine

7-Bromo-4-chloro-3-nitroquinoline (1.9 g, 6.62 mmol) was dissolved in CH ₂ Cl ₂ (20 ml). Added 4methoxybenzylamine (0.85 ml, 6.7 mmol) followed by the addition of NEt ₃ (0.95 ml, 6.7 mmol). The mixture was stirred at room temperature for 4 h, after which it was diluted with CH ₂ Cl ₂ (30 ml), washed with water, washed with brine, dried over Na ₂ SO ₄ and filtered. The resulting solution was evaporated to dryness to give (7-bromo-3-nitroquinolin-4-yl) - (4-methoxybenzyl) amine as a yellow foam (2.5 g, 6.44 mmol, 97%). This material was used in the next step without further purification. (ES, m / z): [M + H] ⁺ = 388.3 / 390.1.

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Stage 4. Obtaining 7-bromo-N ⁴ - (4-methoxybenzyl) quinoline-3,4-diamine

(7-Bromo-3-nitroquinolin-4-yl) - (4-methoxybenzyl) amine (2.5 g, 6.44 mmol) was dissolved in ethanol (60 ml) at room temperature. Tin (II) chloride hydrate (4.8 g, 21.2 mmol) was added in one portion. The mixture was stirred at 65 ° C for 3 h, after which water (50 ml) was added, followed by a saturated aqueous solution of NaHCO ₃ to pH ~ 9. The mixture was extracted with EtOAc (3 x 60 ml) and the combined organic layers were dried over Na ₂ SO ₄ , filtered and concentrated to give 7-bromo-N ⁴ - (4-methoxybenzyl) quinoline-3,4-diamine (1.5 g, 4.2 mmol, 65%). This material was used in the next step without further purification. (ES, m / z): [M + H] + = 357.9 / 360.1.

Example 2. A method of obtaining analogs, where RVR ² is alkyl, acetyl and R ⁵ is aryl, heteroaryl, heterocyclyl or amino.

Scheme 3

Stage 1. Obtaining tert-butyl ((7-bromo-1- (4-methoxybenzyl) -1H-imidazo [4,5-c] quinolin-2-yl) methyl) (ethyl) carbamate

Vos

Not selected

To a solution of 7-bromo-N ⁴ - (4-methoxybenzyl) quinoline-3,4-diamine (0.950 g, 2.65 mmol, example 1) and (tert-butoxycarbonylethylamino) acetic acid (0.600 g, 2.92 mmol) in CH ₂ Cl ₂ (20 ml) was added HATU (1.1 g, 2.65 mmol) and NEt ₃ (0.4 ml, 2.87 mmol). The mixture was stirred at room temperature for 12 hours, after which it was concentrated to remove all volatiles. EtOH (10 ml) was added followed by NEt ₃ (4 ml). The mixture was stirred in an oil bath at 70 ° C for 15 hours and then cooled to room temperature. Most of the volatiles were evaporated in vacuo and the resulting residue was partitioned between CH ₂ Cl ₂ (30 ml) and water (30 ml). The organic layer was washed with additional water (30 ml), brine, dried over Na ₂ SO ₄ , filtered and concentrated. Crude tert-butyl ((7-bromo-1- (4-methoxybenzyl) -1H-imidazo [4,5-c] quinolin-2-yl) methyl) (ethyl) carbamate (0.972 g, 1.85 mmol, yield 70%) was used in the next step without further purification. (ES, m / z): [M + H] ⁺ = 525.1 / 527.4.

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Stage 2. Obtaining N - ((7-bromo-1- (4-methoxybenzyl) -1H-imidazo [4,5-c] quinolin-2-yl) methyl) -Nethylacetamide

To a solution of tert-butyl ((7-bromo-1- (4-methoxybenzyl) -1H-imidazo [4,5-c] quinolin-2-yl) methyl) (ethyl) carbamate (0.972 g, 1.85 mmol) in dioxane (10 ml) was added HCl (10 ml, 4N in dioxane). The mixture was stirred at room temperature for 4 h, after which it was concentrated to remove all volatiles, then dissolved in CH ₂ Cl ₂ (20 ml). The mixture was stirred in an ice water bath for 5 min, then NEt ₃ (0.4 ml, 2.88 mmol) and Ac ₂ O (0.25 ml, 2.64 mmol) were added sequentially. The mixture was further stirred at room temperature for 30 min and then diluted with water (20 ml). The layers were separated and the organic layer was washed with water (20 ml), brine, dried over Na ₂ SO ₄ , filtered and concentrated. Crude N - ((7-6pom-1- (4methoxybenzyl) -1H-imidazo [4,5-c] quinolin-2-yl) methyl) -N-ethylacetamide (0.800 g, 1.71 mmol, 92% yield) was used in the next step without further purification. (ES, m / z): [M + H] + = 467.2 / 469.4.

Stage 3. Obtaining 7-bromo-2 - ((N-ethylacetamido) methyl) -1- (4-methoxybenzyl) -1H-imidazo [4,5c] quinoline-5-oxide

To a solution of N - ((7-bromo-1- (4-methoxybenzyl) -1H-imidazo [4,5-c] quinolin-2-yl) methyl) -N-ethyl acetate (0.800 g, 1.71 mmol) in CH ₂ Cl ₂ (20 ml) was added H ₂ O ₂ (10 ml) and m-chloroperoxybenzoic acid (70% grade, 0.500 g, 0.2 mmol). The mixture was stirred for 15 h at room temperature, after which it was diluted with saturated aqueous NaHCO ₃ solution and extracted with CH ₂ Cl ₂ (3 x 20 ml). The combined organic layers were washed with brine, dried over Na ₂ SO ₄ , filtered and concentrated to give crude 7-bromo-2 - ((N-ethylacetamido) methyl) -1- (4-methoxybenzyl) -1Himidazo [4,5- c] quinoline-5-oxide (0.800 g, 1.65 mmol, 96% yield) as a brownish foam. (ES, m / z): [M + H] + = 483.2 / 485.5.

Stage 4. Obtaining N - ((4-amino-7-bromo-1- (4-methoxybenzyl) -1H-imidazo [4,5-c] quinolin-2-yl) methyl) -N-ethylacetamide

To a solution of 7-bromo-2 - ((N-ethylacetamido) methyl) -1- (4-methoxybenzyl) -1H-imidazo [4,5-c] quinoline 5-oxide (0.800 g, 1.65 mmol) and NH4OH (10 ml) in dichloromethane (20 ml) cooled in an ice bath, p-toluenesulfonyl chloride (0.439 g, 2.3 mmol) in CH ₂ Cl ₂ (10 ml) was added dropwise. The resulting solution was stirred for an additional 30 minutes after the addition was complete. Water (20 ml) was added and the layers were separated. The aqueous layer was extracted with CH ₂ Cl ₂ (30 ml). The combined organic layers were filtered through a pad of Na ₂ SO ₄ and the filtrate was concentrated in vacuo. The residue was triturated with EtOAc / hexanes (1/3) and dried under high vacuum to give N - ((4-amino-7-bromo-1- (4-methoxybenzyl) -1H-imidazo [4,5-c] quinoline- 2-yl) methyl) -N-ethylacetamide as a yellow solid (677 mg, 1.4 mmol, 85% yield). (ES, m / z): [M + H] + = 482.3 / 484.2.

Stage 5. Coupling with aryl bromide.

but. Preparation of N - ((4-amino-1 - (4-methoxybenzyl) -7- (1 H-pyrazol-1-yl) -1 H-imidazo [4,5-c] quinolin-2yl) methyl) -N- ethylacetamide (general method for the condensation reaction of Ullman N heterocycles)

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To a solution of N - ((4-amino-7-bromo-1- (4-methoxybenzyl) -1H-imidazo [4,5-c] quinolin-2-yl) methyl) -Nethylacetamide (25 mg, B 52 mol) in dry DMSO (2 ml), N-heterocycle (2 eq., pyrazole in the above example) was added, followed by CuI (25 mg, 2 eq.) and Na ₂ CO ₃ (30 mg, 4 eq.). The mixture was degassed, N, N'-dimethylethylenediamine (20 mg, 3 eq.) Was added and the mixture was stirred at 120 ° C for 2 h. The cooled mixture was diluted with EtOAc, filtered and the solvent was evaporated. The resulting crude N - ((4-amino-1- (4-methoxybenzyl) -7- (1H-pyrazol-1-yl) -1H-imidazo [4,5-c] quinolin-2-yl) methyl) -N α-ethylacetamide was used in the deprotection step without further purification, or

5b. Alternative preparation for setting aryl groups in place of N-heterocycles: general procedure for a Suzuki cross-coupling reaction of acylboronic acids and esters

To a solution of N - ((4-amino-7-bromo-1- (4-methoxybenzyl) -1H-imidazo [4,5-c] quinolin-2-yl) methyl) -Nethylacetamide (25 mg, 52 mol) was added aryl (heteroaryl) boric acid (or aryl boronate ester) (2 eq.), then Pd (dppf) Cl ₂ -CH ₂ Cl ₂ (5 mg) and an aqueous solution of K ₂ CO ₃ (1 ml, 2M aqueous). The mixture was heated in a Biotage InitiatoR microwave reactor at 120 ° C for 10 min. The organic layer was diluted with EtOAc and separated, and the aqueous layer was washed with EtOAc. The combined organic phases were filtered, evaporated and the product residue was used in the next step without further purification.

Step 6. General deprotection procedure - preparation of N - ((4-amino-7-aryl (heteroaryl) -1H-umidazo [4,5-c] quinolin-2-yl) methyl) -N-ethylacetamide

The crude product from the previous step was dissolved in TFA (2 ml) and stirred at 70 ° C for 1 hour, while monitoring the reaction with LC / MS. Once the protecting group PMB was completely cleaved as indicated by LC / MS, the solvent was evaporated and the residue was purified by HPLC to give N - ((4-amino-7-aryl (heteroaryl) -1H-umidazo [4,5-c] quinoline- 2-yl) methyl) -N-ethylacetamide as trifluoroacetic salt.

The compounds shown in table. 2 were prepared according to the synthesis methods described above.

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table 2

Compound NAME [M + HJ + 165 T4 - [(4-amino-7-phenyl-1H-imidazo [4,5c] quinoline-2-y1) methyl] -M-ethylacetamide 360.2 166 N- [(4-amino-7-pyridin-3-yl-1Nimidazo [4,5-c] quinolin-2-yl) methyl] -Nethylacetamide 361.2 167 N- [(4-amino-7-pyrid in-4-yl-1Nimidazo [4,5-c] quinolin-2-yl) methyl] -Nethylacetamide 361.2 168 N- [[4-amino-7- (2,5-dimethylpyrazol-3-yl) 1 H-imidazo [4,5-c] quinolin-2-yl] methyl] -Nethylacetamide 378.2 169 N- [[4-amino-7- (1-propylpyrazol-4-yl) -1Nimidazo [4,5-c] quinolin-2-yl] methyl] -Nethylacetamide 392.2 170 N- [(4-amino-7-propan-2-yl-1 H-imidazo [4.5 s] quinolin-2-yl) methyl] A-ethylacetamide 326.2 171 A [(4-amino-7-ethyl-1 H-imide azo [4.5 s] quinolin-2-yl) methyl] A-ethylacetamide 312.2 172 H - [(4-amino-7-propyl-1H-imidazo [4,5c] quinolin-2-yl) methyl] A-ethylacetamide 326.2 173 A [(4-amino-7-cyclopentyl-1Nimidazo [4,5-c] quinolin-2-yl) methyl] -Nethylacetamide 352.2 175 N- [[4-amino-7- (2-fluoropyridin-3-yl) -1Nimidazo [4,5 -c] quinolin-2-yl] methyl] -Nethylacetamide 379.2 177 N - [(4-aMHHO-7-6poM-l H-imide azo [4,5c] quinolin-2-yl) methyl] -H-ethylacetamide 362.1 179 H - [(4-amino-7-chloro-1H-imidazo [4,5c] quinolin-2-yl) methyl] -No-ethylacetamide 318.1 180 H - [(4-amino-7-pyrazol-1-yl-1Nimidazo [4,5-c] quinolin-2-yl) methyl] -Nethylacetamide 350.2 182 M- [1- (4-amino-7-bromo-1H-imidazo [4,5c] quinolin-2-yl) ethyl] A-ethylacetamide 376.1

Example 3. An example of a method for producing analogs, where R ¹ is H, R ² is alkyl and R ⁵ is aryl, heteroaryl, heterocyclyl or amino.

The following synthetic schemes were used to prepare compound 101 and related analogs, compounds 463, 465 and 468. Additional analogs of these compounds can be obtained, for example, by replacing the hydrogen of the secondary amino group of compound 101 with a number of additional substituents, for example, by acylation and sulfonylation using transformations, known to those skilled in the art as described in Example 4. Additional analogs thus obtained include, for example, compounds 105, 109, 110, 112 (described in Example 4a), 114, 118, 121, 123, 124, 125 , 126, 133, 379, 383, 395, 406, 410 and 411.

Example 3a. Preparation of 2 - [(ethylamino) methyl] -7- (1H-pyrazol-3-yl) -1H-imidazo [4,5-c] quinoline-4amine hydrochloride (compound 101).

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Scheme 4

Stage 1. tert-Butyl-No. - [[(4-amino-7-bromoquinolin-3-yl) carbamoyl] methyl] -N-ethyl carbamate

A solution of 7-bromoquinoline-3,4-diamine (375 g, 1.58 mol, 1.00 eq .; Bioorg. Med. Chem. Lett, 2012 , 22, 285) in ethyl acetate (6 L). To the solution was added pyridine (623 g, 7.88 mol, 5.00 eq.), 2 - [(tert-butoxy) carbonyl] (ethyl) aminoacetic acid (480 g, 2.36 mol, 1.50 eq.) and T ₃ P (2004, 3.15 mol, 2 eq.). The resulting solution was stirred for 16 hours at room temperature. The resulting mixture was washed with aqueous sodium hydroxide solution (3x10 L) and brine (2x10 L). The resulting mixture was concentrated in vacuo. This gave 592 g (89%) tert-butylU - [[(4-amino-7-bromoquinolin-3-yl) carbamoyl] methyl] -Y-ethyl carbamate as a yellow solid.

Stage 2. tert-Butyl-N - [(7-bromo-1H-imidazo [4,5-c] quinolin-2-yl] methyl) -N-ethyl carbamate

A solution of tert-butyl-U - [[(4-amino-7-bromoquinoline-3yl) carbamoyl] methyl] A-ethyl carbamate (592 g, 1.40 mol, 1.00 eq.) In ethanol (6 L). Sodium hydroxide (558 g, 13.95 mol, 10.00 ex) was added to the solution. The resulting solution was stirred for 1 hour at 80 ° C in an oil bath. The resulting mixture was concentrated in vacuo and diluted with 5 L of DCM. The resulting mixture was washed with saturated brine (5x10 L), dried over anhydrous sodium sulfate, and concentrated in vacuo. This reaction gave 400 g (71%) tert-butyl-N - [(7-bromo-1Himidazo [4,5-c] quinolin-2-yl] methyl) -N-ethyl carbamate as a yellow solid.

Step 3. 7-Bromo-2 - ([[(tert-butoxy) carbonyl] (ethyl) amino] methyl) -1H-imidazo [4,5-c] quinoline-5ium-5-olate

A solution of tert-butyl-N - ([7-bromo-1H-imidazo [4,5-c] quinolin-2-yl] methyl) -N - 86 037780 ethyl carbamate (400 g, 986.95 mmol, 1.00 equiv.) In dichloromethane (6 L). CPBA (342 g, 1.98 mol, 2.00 ex) was added to the solution. The resulting solution was stirred for 16 hours at room temperature. The resulting mixture was washed with aqueous sodium carbonate solution (3x10 L), dried over anhydrous sodium sulfate, and concentrated in vacuo. The residue was washed with 5 L of ethyl acetate. The solids were collected by filtration. As a result, 218.6 g (in pure form) and 341 g (crude) 7-bromo-2 ([[(tert-butoxy) carbonyl] (ethyl) amino] methyl) -1H-imidazo [4,5-c ] quinolin-5-ium-5-olate as a light yellow solid.

Step 4. tert-Butyl-N - [(4-amino-7-bromo-1H-imidazo [4,5-c] quinolin-2-yl] methyl) -N-ethyl carbamate

A solution of 7-bromo-2 - ([[(tert-butoxy) carbonyl] (ethyl) amino] methyl) -1H-imidazo [4,5-c] quinolin-5-ium -5-olate (214 g, 518.31 mmol, 1.00 eq.) In dichloromethane (3 L) and NH ₃ -H2O (1 L). To the solution was then added 4-methylbenzene-1-sulfonyl chloride (194 g, 1.037 mol, 2.00 eq.). The resulting solution was stirred for 6 hours at room temperature. The solids were collected by filtration, washed with DCM (3x2 L) and dried. As a result, 188 g (86%) of tert-butyl-N - [(4-amino-7-bromo-1H-imidazo [4,5-c] quinolin-2-yl] methyl) N-ethyl carbamate were obtained in the form of light- yellow solid.

Step 5. tert-Butyl-N - [[4-amino-7- (1H-pyrazol-3-yl) -1H-imidazo [4,5-c] quinolin-2-yl] methyl] -Nethyl carbamate

A solution of tert-butyl-N - ([4-amino-7-bromo-1H-imidazo [4,5-c] quinolin-2-yl] methyl) -N-ethyl carbamate (156 g, 371.16 mmol, 1.00 eq.) In 1,4-dioxane / H2O (3 / 0.3 L). To the resulting solution was added Cs ₂ CO ₃ (363 g, 1.11 mol, 3.00 eq), 3- (tetramethyl-1,3,2-dioxaborolan-2-yl) -1H-pyrazole (144 g, 742, 12 mmol, 2.00 eq.) And Pd (dppf) Cl ₂ dichloromethane complex (30 g, 36.72 mmol, 0.10 eq.). The resulting solution was stirred for 24 h at 100 ° C in an oil bath. The resulting solution was cooled to room temperature and diluted with 3 L of H2O. The solid was collected by filtration and loaded onto a silica gel column with dichloromethane / methanol (20/1). As a result, 76.3 g (50%) of tert-butyl-N - [[4-amino-7- (1H-pyrazol-3-yl) -1H-imidazo [4,5-c] quinoline-2- yl] methyl] -N-ethyl carbamate as a yellow solid.

Stage 6. Compound 101

A solution of tert-butyl-N - [[4-amino-7- (1H-pyrazol-3-yl) 1H-imidazo [4,5-c] quinolin-2-yl] methyl] - N-ethyl carbamate (75.6 g, 185.54 mmol, 1.00 eq.) In 1,4-dioxane / HCl (2 L, 4M). The resulting solution was stirred for 16 hours at room temperature. The resulting mixture was concentrated in vacuo. The residue was slurried in 2 L of dichloromethane and the solid was collected by filtration. This procedure was repeated three times. The collected solid was then dried to give 75.7 g of (crude) 2 - [(ethylamino) methyl] -7- (1H-pyrazol-3-yl) -1Himidazo [4,5-c] quinolin-4-amine hydrochloride in the form of a yellow solid. LC / MS [M + H] 308.3.

LC / MS conditions: Column: VEN C18 2.1 x 50 mm; mobile phase A: water with 0.05% TFA; Mobile Phase B: Acetonitrile with 0.05% TFA; temperature: 50 ° C; gradient: 2-98% B over 1.7 minutes; flow rate: 0.8 ml / min; LC RT = 0.46 min. ¹ H NMR (400 MHz, METHANOL-d ₄ ) δ 8.11 (d, J = 8.3 Hz, 1H), 8.04 (s, 1H), 7.84 - 7.77 (m, 1H) , 7.72 (s, 1H), 6.79 (d, J = 2.1 Hz, 1H), 4.13 (s, 2H), 2.80 (q, J = 7.2 Hz, 2H) , 1.22 (t, J = 7.2 Hz, 3H).

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Example 3b. Preparation of 1-ethyl-2 - [(ethylamino) methyl] -7- (1H-pyrazol-5-yl) -1H-imidazo [4,5-c] quinolin-4-amine (compound 464). Scheme 5

Stage 1. 7-Bromo-Nethyl-3-nitroquinoline-4-amine

A solution of 7-bromo-4-chloro-3-nitroquinoline (20 g, 62.61 mmol, 1 eq., 90%) in dichloromethane (300 ml) was placed in a 500 ml round bottom flask. Then ethanamine (4.23 g, 93.91 mmol, 1.5 eq.) And triethylamine (19.0 1 g, 187.83 mmol, 3 ex) were added. The resulting solution was stirred for 1 hour at room temperature. The reaction was then quenched by the addition of water. The resulting solution was extracted three times with 100 ml of dichloromethane, and the organic layers were combined. The solution was dried over anhydrous sodium sulfate and concentrated. This gave 20 g of 7-bromo-N-ethyl-3-nitroquinolin-4-amine as a yellow crude solid.

A solution of 7-bromo-Nethyl-3-nitroquinoline-4amine (20 g, 67.54 mmol, 1 eq.) In CH ₃ CN (500 ml) was placed in a 1000 ml round bottom flask. Pt / C (3 g, 15.38 mmol, 0.23 eq.) Was added to the solution. The resulting solution was degassed and backfilled with H ₂ . The resulting solution was stirred for 24 hours at room temperature. The solids were filtered off. The filtrate was concentrated to give 19.7 g of 7-bromo-N ⁴ -ethylquinoline-3,4-diamine as a yellow crude oil. LC / MS: (ES, m / z): [M + H] + = 226.1.

Stage 3.tert-Butyl- (2 - ((7-bromo-4- (ethylamino) quinolin-3-yl) amino) -2-oxoethyl) (ethyl) carbamate

A solution of 7-bromo-N ^4- ethylquinoline-3,4-diamine (10 g, 35.70 mmol, 1 eq, 95%) and 2 - [[(tert-butoxy) carbonyl] ( ethyl) amino] acetic acid (10 0.88 g,

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53.54 mmol, 1.5 eq.) In dichloromethane (500 ml). This was followed by the addition of HATU (16.2 9 g, 42.83 mmol, 1.2 eq.) And DIEA (15.34 g, 107.09 mmol, 3 eq.). The resulting solution was stirred for 3 hours at room temperature. The reaction was then quenched by the addition of water. The resulting solution was extracted three times with 100 ml of dichloromethane, and the organic layers were combined. The solution was dried over anhydrous sodium sulfate and concentrated. This gave 40 g of (crude) tert-butylNo ([[7-bromo-4- (ethylamino) quinolin-3-yl] carbamoyl] methyl) -No ethyl carbamate as a yellow oil.

Stage 4. tert-Butyl-N ([7-bromo-1-ethyl-1H-imidazo [4,5-c] quinolin-2-yl] methyl) -No ethyl carbamate

1000 ml was placed a solution of tert-butyl-N [[(3-amino-7In a round-bottom flask with a volume of bromoquinolin-4-yl) (ethyl) carbamoyl] methyl] -No ethyl carbamate (40 g, 88.62 mmol, 1 eq.) In ethanol (500 ml) and water (20 ml). To the resulting solution was added sodium hydroxide (35, 45 g, 886.20 mmol, 10 eq). The resulting solution was stirred for 2 h at 100 ° C. The resulting mixture was concentrated. The residue was diluted with water. The resulting solution was then extracted with ethyl acetate and the organic layers were combined. The solution was dried over anhydrous sodium sulfate and concentrated. This gave 34.8 g (crude) tert-butyl-N ([7-bromo-1-ethyl-1H-imidazo [4,5-c] quinolin-2-yl] methyl) -No ethyl carbamate as a yellow oil ... LC / MS: (ES, m / z): [M + H] + = 433.3.

Step 5. 7-Bromo-2 - ([[(tert-butoxy) carbonyl] (ethyl) amino] methyl) -1-ethyl-1 H-imidazo [4,5-c] quinoline-5-ium-5- olat

In a round bottom flask with a volume of 1000 ml was placed tert-butyl-N ([7-bromo-1-ethyl-1H-imidazo [4,5c] quinolin-2-yl] methyl) -No ethyl carbamate (6 g, 13.85 mmol, 1 eq.) In dichloromethane (300 ml). This was followed by the addition of m-CPBA (3.58 g, 20.77 mmol, 1.5 eq) in portions. The resulting solution was stirred for 2 hours at room temperature. The reaction mixture was quenched by the addition of 150 ml of H ₂ O. The resulting solution was extracted with DCM and the organic layers were combined. The solution was dried over anhydrous sodium sulfate and concentrated. As a result, 7.5 g of 7-bromo-2 - ([[(tert-butoxy) carbonyl] (ethyl) amino] methyl) -1-ethyl-1 H-imidazo [4,5-c] quinoline-5-ium- 5-olate in the form of a crude yellow solid.

Stage 6. tert-Butyl-N - ([4-amino-7-bromo-1-ethyl-1H-imidazo [4,5-c] quinolin-2-yl] methyl) -N-ethyl carbamate

7-bromo-2 - ([[(tert-butoxy) carbonyl] (ethyl) amino] methyl) -1-ethyl-1H-imidazo [4,5-c] quinoline-5-ium -5-olate (6 g, 13.35 mmol, 1 eq., Crude) in dichloromethane (250 ml) and NH ₃ H ₂ O (5 ml, 20 mmol). This was followed by the addition of a solution of 4-methylbenzene-1-sulfonyl chloride (5.09 g, 26.71 mmol, 2 eq.) In dichloromethane (10 mL) dropwise with stirring. The resulting solution was stirred for 2 hours at room temperature. The reaction was then quenched by the addition of water. The resulting solution was extracted with dichloromethane and the organic layers were combined. The solution was dried over anhydrous sodium sulfate and concentrated. As a result, 2 g of tert-butyl-N - ([4-amino-7-bromo-1-ethyl-1H-imidazo [4,5-c] quinolin-2-yl] methyl) N-ethyl carbamate was obtained in the form of a white solid substances. LC / MS: (ES, m / z): [M + H] ⁺ = 448.1, 1H-NMR: (400 MHz, CD ₃ OD) δ 8.04-8.02 (m, 1H), 7 , 85-7.84 (m, 1H), 7.47-7.45 (m, 1H), 4.84-4.82 (m, 2H), 4.67 (s, 2H), 3.35 (m, 2H), 1.5-1.29 (m, 12H), 1.09-0.89 (m, 3H).

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Step 7. tert-Butyl-N - [[4-amino-1-ethyl-7- (1H-pyrazol-5-yl) -1H-imidazo [4,5-c] quinolin-2-yl] methyl] - N-ethylcarbamate

A solution of tert-butyl-N - ([4-amino-7-bromo-1-ethyl-1H-imidazo [4,5-c] quinoline-2 -yl] methyl) N-ethyl carbamate (150 mg, 0.30 mmol, 1 eq., 90%), 5- (4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl) -1Npyrazole (116.8 mg, 0.60 mmol, 2 eq.) In dioxane (25 ml) under N2. To the solution were added Cs ₂ CO ₃ (294.3 mg, 0.90 mmol, 3 eq.) And Pd (dppf) Cl ₂ (11.0 mg, 0.02 mmol, 0.05 eq.). The resulting solution was stirred for 24 h at 100 ° C. The reaction was then quenched by the addition of water. The resulting solution was extracted with ethyl acetate. The organic layers were combined and dried over anhydrous sodium sulfate. The resulting mixture was concentrated. The residue was applied to a silica gel column and eluted with dichloromethane / methanol (10: 1). This led to the production of 100 mg (76%) tert-butyl-N - [[4-amino-1-ethyl-7 (1H-pyrazol-5-yl) -1H-imidazo [4,5-c] quinoline-2 -yl] methyl] -N-ethyl carbamate as a yellow solid.

Stage 8. Synthesis of compound 465

Tert-butyl-N - [[4-amino-1-ethyl-7- (1H-pyrazol-5yl) -1H-imidazo [4,5-c] quinolin-2-yl] methyl ] -N-ethyl carbamate (100 mg, 0.23 mmol, 1 eq.) In dichloromethane (20 ml). This was followed by the addition of HCl in dioxane (1.5 ml) dropwise with stirring. The resulting solution was stirred for 1 hour at room temperature. The resulting mixture was concentrated. The crude product was purified by preparative HPLC with the following conditions. Column: SunFire C18 OBD Prep. Column, 100 A, 5 microns, 19 mm x 250 mm; Mobile phase A: water (0.05% TFA), mobile phase B: ACN; flow rate: 25 ml / min; gradient: 5% B to 30% B in 8 minutes; 254/210 nm; Rt: 7.03 min. As a result, 18 mg (13.77%) of 1-ethyl-2 - [(ethylamino) methyl] -7- (1H-pyrazol-5-yl) -1Nimidazo [4,5-c] quinolin-4-amine were obtained; bis (trifluoroacetic acid) as a white solid. LC Methods: Column: Express C18 2.1 mm x 50 mm, 2.7 μm particles; mobile phase A: water with 0.05% TFA; Mobile Phase B: Acetonitrile with 0.05% TFA; temperature: 40 ° C; gradient: 5% B to 100% B over 2 min, then held at 100% B for 0.75 min; flow rate: 0.8 ml / min. Retention time LC: 0.81 min. LC / MS: (ES, m / z): [M + H] + = 336, 3, 1H-NMR: (300 MHz, CD ₃ OD, ppm) δ 8.38-8.35 (d , J = 8.7 Hz, 1H), 8.22 (m, 1H), 8.12-8.09 (m, 1H), 7.80-7.79 (d, J = 2.8 Hz, 1H), 6.89-6.86 (m, 1H), 4.76-4.69 (m, 4H), 3.453.38 (m, 2H), 1.63 (t, J = 7.2 Hz , 3h), 1.48 (t, J = 72 Hz, 3H).

Example 3c. Preparation of 1-ethyl-2 - ((ethylamino) methyl) -7- (1H-pyrazol-1-yl) -1H-imidazo [4,5-c] quinolin-4-amine (compound 467).

Scheme 6

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Stage 1. Synthesis of 1-ethyl-2 - [(ethylamino) methyl] -7- (1H-pyrazol-1-yl) -1H-imidazo [4,5-c] quinoline-4 amine

In a 50 ml round bottom flask, purged and maintained in an inert nitrogen atmosphere, was placed tert-butyl-N - ([4-amino-7-bromo-1-ethyl-1 H-imidazo [4,5-c] quinoline-2 -yl] methyl) -N-ethyl carbamate (150 mg, 0.33 mmol, 1 eq.), 1H-pyrazole (45.6 mg, 0.67 mmol, 2 eq.) and K ₃ PO ₄ (213.0 mg, 1.00 mmol, 3 eq.) in DMF (5 ml). Then (1S, 2S) -cyclohexane-1,2-diamine (15.3 mg, 0.13 mmol, 0.4 equivalents) and CuI (25.5 mg, 0.13 mmol, 0.4 equivalents) were added. The resulting solution was stirred for 24 h at 100 ° C under N2. The reaction mixture was then diluted with water. The resulting solution was extracted three times with 50 ml of ethyl acetate and the organic layers were combined. The solution was dried over anhydrous sodium sulfate and concentrated. The residue was applied to a silica gel column with dichloromethane / methanol (10: 1). As a result, we obtained 100 mg (80.20%) of 1-ethyl-2 - [(ethylamino) methyl] -7- (1H-pyrazol-1-yl) -1H-imidazo [4,5-c] quinolin-4-amine in the form of a white solid. LC / MS: (ES, m / z): [M + H] + = 436.2.

Stage 2. Synthesis of compound 468

A solution of tert-butyl-N - [[4-amino-1-ethyl-7- (1Hpyrazol-1-yl) -1H-imidazo [4,5-c] quinolin-2-yl] methyl] -N-ethyl carbamate (100 mg, 0.23 mmol, 1 eq.) in DCM (10 ml). This was followed by the addition of HCl in dioxane (1.5 ml) dropwise with stirring. The resulting solution was stirred for 1 hour at room temperature. The resulting mixture was concentrated. The crude product was purified by preparative HPLC with the following conditions. Column: SunFire C18 OBD Prep Column, 100A, 5μm, 19mm x 250mm; Mobile Phase A: Water (0.05% TFA) Mobile Phase B: ACN; flow rate: 25 ml / min; gradient: 5% B to 30% B in 10 minutes; 254/210 nm; Rt: 9.20 min. As a result, 33.5 mg (25.64%) 1-ethyl-2 - ((ethylamino) methyl) -7- (1H-pyrazol-1-yl) -1H-imidazo [4,5-c] quinoline-4- amine as a white solid. LC / MS conditions: Column: Kinetex EVO, 3.0 mm x 50 mm, 2.6 μm particles; mobile phase A: water with 5 mM ammonium bicarbonate; mobile phase B: acetonitrile; temperature: 40 ° C; gradient: 10% B to 95% B over 2 min, then held at 95% B for 0.79 min; flow rate: 1 ml / min. Retention time LC: 1.01 min. LC / MS: (ES, m / z): [M + H] + = 336.3; 1H-NMR: 1HNMR (300 MHz, CD3OD) δ 8.46-8.40 (m, 2H), 8.25-8.24 (m, 1H), 8.10-8.06 (m, 1H) , 7.84-7.83 (m, 1H) 6.64-6.63 (m, 1H), 4.72 (s, 2H), 4.70-4.63 (m, 2H), 3, 54-3.08 (m, 2H), 1.62 (t, J = 6.0 Hz, 3H), 1.48 (t J = 9.0 Hz, 3H).

Example 3d. Preparation of 2- [4-amino-2 - [(ethylamino) methyl] -7- (1H-pyrazol-5-yl) -1H-imidazo [4,5-c] quinolin-1-yl] ethan-1-ol (compound 462).

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Scheme 7

Stage 1. X [2- (benzyloxy) ethyl] -7-bromo-3-nitroquinoline-4-amine

To a stirred mixture of 7-bromo-4-chloro-3-nitroquinoline (20 g, 69.57 mmol, 1 eq.) And 2 (benzyloxy) ethan-1-amine (12.0 g, 79.30 mmol, 1, 14 eq.) In DCM (400 ml) was added TEA (10.6 g, 104.35 mmol, 1.50 eq.) At room temperature under nitrogen atmosphere. The resulting mixture was stirred for 16 hours at room temperature under a nitrogen atmosphere. The resulting mixture was concentrated in vacuo. This gave X [2- (benzyloxy) ethyl] -7-bromo-3-nitroquinolin-4-amine (30 g) as a yellow crude solid. LC / MS: (ES, m / z): [M + H] + = 402.2 / 404, 2.

Stage 2. N ⁴ - [2- (benzyloxy) ethyl] -7-bromoquinoline-3,4-diamine

To a solution of X [2- (benzyloxy) ethyl] -7-bromo-3-nitroquinolin-4-amine (30 g, 74.58 mmol, 1 eq.) In CH ₃ CN (400 ml) was added Pt / C (2 , 9 g, 14.87 mmol, 0.20 eq.) Under nitrogen in a 1000 mL round bottom flask. The mixture was hydrogenated at room temperature for 16 hours under a hydrogen atmosphere using a hydrogen balloon, filtered through a celite pad, and concentrated under reduced pressure. The residue was purified by column chromatography on silica gel eluting with CH2Cl2 / MeOH (20: 1) to give N4- [2- (benzyloxy) ethyl] -7-bromoquinoline-3,4-diamine (23.4 g, 84.28%) as a yellow solid. LC / MS: (eS, m / z): [M + H] ⁺ = 372.3 / 374.2.

Stage 3. tert-Butyl-N - [[(4 - [[2- (benzyloxy) ethyl] amino] -7-bromoquinolin-3-yl) carbamoyl] methyl] N-ethyl carbamate

OBn

To a stirred mixture of N4- [2- (benzyloxy) ethyl] -7-bromoquinoline-3,4-diamine (14.9 g, 40.03 mmol, 1 eq.) And 2 - [[(tert-butoxy) carbonyl] (ethyl) amino] acetic acid (8.9 g, 44.03 mmol, 1.1 eq.) in DCM (500 ml) was added HATU (18.3 g, 48.03 mmol, 1.2 eq.) and DIEA (10.3 g, 79.69 mmol, 1.99 equiv.) At

- 92 037780 room temperature. The resulting mixture was stirred for 16 hours at room temperature under a nitrogen atmosphere. The resulting mixture was concentrated in vacuo. The result was tert-butyl-N [[(4 - [[2- (benzyloxy) ethyl] amino] -7-bromoquinolin-3-yl) carbamoyl] methyl] -N-ethyl carbamate (24 g, 107.56%) in the form of a red crude oil. The crude product was used directly in the next step without further purification. LC / MS: (ES, m / z): [M + H] ⁺ = 557.2 / 559.2.

Step 4. tert-Butyl-N - ([1 - [2- (benzyloxy) ethyl] -7-bromo-1 H-imidazo [4,5-c] quinolin-2-yl] methyl) -N ethyl carbamate

A mixture of tert-butyl-N - [[(4- [[2- (benzyloxy) ethyl] amino] -7-bromoquinolin-3-yl) carbamoyl] methyl] -Nethyl carbamate (23 g, 41.26 mmol, 1 eq. ) and NaOH (3.3 g, 82.51 mmol, 2 eq.) in MeOH (250 ml) were stirred for 16 h at 65 ° C. The resulting mixture was concentrated in vacuo. The residue was purified by column chromatography on silica gel eluting with PE / EtOAc (1.5: 1) to give tert-butyl-N - ([1- [2 (benzyloxy) ethyl] -7-bromo-1H-imidazo [4.5- c] quinolin-2-yl] methyl) -N-ethyl carbamate (17.4 g, 78.18%) as a light yellow solid. LC-MS: (ES, m / z): [M + H] ⁺ = 539.2 / 541.2 H-NMR: 1H NMR (400 MHz, DMSO-d6) δ 9.23 (s, 1H), 8.42-8.31 (m, 2H), 7.75 (dd, J = 8.9, 2.2 Hz, 1H), 7.14 (dd, J = 5.0, 2.0 Hz, 3H), 7.00 (dd, J = 6.7, 2.9 Hz, 2H), 4.95 (s, 2H), 4.83 (s, 2H), 4.39 (s, 2H), 3.86 (d, J = 6.0 Hz, 2H), 2.51 (p, J = 1.8 Hz, 2H), 1.38 (d, 7 = 42.6). Hz, 9H), 0.99 (t, J = 7.0 Hz, 3H).

Step 5. tert-Butyl-N - ([1 - [2- (benzyloxy) ethyl] -7-bromo-1 H-imidazo [4,5-c] quinolin-2-yl] methyl) -N ethyl carbamate

A mixture of 1 - [2- (benzyloxy) ethyl] -7-bromo-2 - ([[(tert-butoxy) carbonyl] (ethyl) amino] methyl) -1 H-imidazo [4,5-c] quinoline-5 -ium-5-olate (2.1 g, 3.78 mmol, 1 eq.) and mCPBA (1.0 g, 5.67 mol, 1.5 eq.) in DCM (35 ml) were stirred for 4 h at room temperature in a nitrogen atmosphere. The resulting mixture was concentrated in vacuo. The residue was purified by column chromatography on silica gel eluting with CH ₂ Cl ₂ / MeOH (20: 1) to give tert-butyl N - ([1- [2- (benzyloxy) ethyl] -7-bromo-1H-imidazo [4 , 5-c] quinolin2-yl] methyl) -N-ethyl carbamate (1.15 g, 56.39%) as a yellowish white solid. LC / MS: (ES, m / z): [M + H] ⁺ = 555.2 / 557.2.

Step 6. tert-Butyl-N - ([4-amino-1- [2- (benzyloxy) ethyl] -7-bromo-1H-imidazo [4,5-c] quinoline-2yl] methyl) -N-ethyl carbamate

1- [2- (benzyloxy) ethyl] -7-bromo-2 - ([[(tert-butoxy) carbonyl] (ethyl) amine K of a stirred mixture but] methyl) -1H-imidazo [4,5-c] quinoline- 5-ium-5-olate (1.15 g, 2.07 mmol, 1 eq.) And NH ₄ OH (5 ml) in DCM (20 ml) was added TsCl (0.8 g, 4.20 mol, 2 , 03 equiv.) At room temperature. The resulting mixture was stirred for 1 hour at room temperature. The resulting mixture was concentrated in vacuo. The residue was purified by column chromatography on silica gel eluting with CH ₂ Cl ₂ / McOH (30: 1) to give tert-butyl N - ([4-amino-1- [2- (benzyloxy) ethyl] -7-bromo-1H α-imidazo [4,5-c] quinolin-2-yl] methyl) -Nethyl carbamate (775 mg, 67.51%) as an off-white solid. LC-MS: (ES, m / h): [M + H] + = 554.2 / 556.2.

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Stage 7. Seret-Butyl-N - ([4-amino-1- [2- (benzyloxy) etul] -7- (1H-purazol-5-yl) -1H-imidazo [4,5c] quinoline-2- yl] methyl) -N-ethylcarbamate

To a solution of tert-butyl-N - ([4-amino-1- [2- (benzyloxy) etul] -7-bromo-1H-imidazo [4,5-c] quinolin-2yl] methyl) -N-ethyl carbamate ( 350 mg, 0.63 mmol, 1 eq.) And 5- (4,4,5,5-tetramethyl-1,3,2-dioxaborolane-2yl) -1H-pyrazole (245.0 mg, 1.26 mmol , 2.00 eq.) In dioxane (8 ml) and H ₂ O (0.8 ml) were added K ₂ CO ₃ (261.7 mg, 1.89 mmol, 3 eq.) And Pd (dppf) Cl ₂ CH ₂ Cl ₂ (103.1 mg, 0.13 mmol, 0.2 eq.) Under nitrogen. After stirring for 20 hours at 90 ° C. under nitrogen atmosphere, the resulting mixture was concentrated under reduced pressure. The residue was purified by column chromatography on silica gel eluting with CH ₂ Q ₂ / MeOH (20: 1) to give tert-butyl-N - ([4-amino-1- [2- (benzyloxy) ethyl] -7- (1H- nyrazol-5-yl) 1H-imidazo [4,5-c] quinolin-2-yl] methyl) -N-ethylcarbamate (250 mg, 73.12%) as a brown solid. LC / MS: (ES, m / z): [M + H] + = 542.2.

Stage 8. Compound 463

A mixture of ret-butyl-N - ([4-amino-1- [2- (benzyloxy) ethyl] -7- (1H-pyrazol-5-yl) -1H-imidazo [4,5-c] quinoline-2- yl] methyl) -N-ethyl carbamate (210 mg, 390 mmol, 1 eq.) in HCl in dioxane (10 ml, 4 mol / L) was stirred for 24 h at 60 ° C. The resulting mixture was concentrated in vacuo. The crude product was purified by preparative HPLC under the following conditions (column: XBridge Prep C18 OBD column, 5 μm, 19 * 150 mm; mobile phase A: water (10 mmol / L NH4HCO3), mobile phase B: ACN; flow rate: 25 ml / min; gradient: 12% B to 26% B in 10 min; 254/210 nm; Rt: 8.63 min), which gave 2- [4-amino2 - [(ethylamino) methyl] -7- (1H- pyrazol-5-yl) -1H-imidazo [4,5-c] quinolin-1-yl] ethan-1-ol (8.1 mg, 5.95%) as a white solid. LC methods: Column: Shim-pack XR-ODS 3.0 mm x 50 mm, 2.2 μm particles; mobile phase A: water with 0.05% TFA; Mobile Phase B: Acetonitrile with 0.05% TFA; temperature: 40 ° C; gradient: 0% B to 95% B over 2 min, then held at 95% B for 0.7 min; flow rate: 1.5 ml / min. Retention time LC: 0.76 min. LC / MS: (ES, m / z): [M + H] + = 352.3 H-NMR: 1H NMR (300 MHz, DMSO-d ₆ ) δ 13.15 (m, 1H), 8.16 -7.95 (m, 2H), 7.89-7.48 (m, 2H), 6.80 (s, 1H), 6.54 (s, 2H), 5.57 (s, 1H), 4.75 (d, J = 6.3 Hz, 2H), 4.05 (s, 2H), 3.91 (t, J = 5.3 Hz, 2H), 2.61 (q, J = 7 , 1 Hz, 2H), 1.04 (t, J = 7.0 Hz, 3H).

Compounds in the table. 3 were obtained according to the methods outlined above.

Table 3

Compound NAME LC / MS [M ⁺ + H] LC RT LC Method 468 1-ethyl-2 - [(ethylamino) methyl] -7 (thiophen-2-yl) -1H-imidazo [4,5c] quinolin-4-amine 352.1 1.29 minutes BUT 471 2- {4-amino-2 [(ethylamino) methyl] -7- (thiophen2-yl) -1H-imidazo [4,5c] quinolin-1-yl (ethan-1-ol 368.1 1.12 minutes IN 477 2- [4-amino-2 [(ethylamino) methyl] -7- (1Hpyrazol-1-yl) -1 H-imidazo [4.5 s] quinolin-1-yl] ethane-1-ol 352.0 0.92 minutes IN

LC Methods: A: Column: Kinetex EVO, 3.0 mm x 50 mm, 2.6 μm particles; mobile phase A: water with 5 mM ammonium bicarbonate; mobile phase B: acetonitrile; temperature: 40 ° C; gradient: 10% B to 95% B over 2 min, then held at 95% B for 0.79 min; flow rate: 1 ml / min. B: Column: Kinetex EVO, 3.0 mm x 50 mm, 2.6 μm; mobile phase A: water with 0.03% NH3H2O; mobile phase B: acetonitrile; temperature: 40 ° C; gradient: 10% B to 95% B over 2 min, then held at 95% B for 0.60 min; flow rate: 1.2 ml / min.

Example 4. An example of a method for producing analogs, where RVR ² is H, alkyl, acetyl, sulfonyl, carbonyl, amido, carboxy and R ⁵ is aryl, heteroaryl, heterocyclyl, or amino.

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Example 4a. An example of a procedure for the formation of acetamide

Preparation of N- [[4-amino-7- (1 H-pyrazol-3-yl) -1 H-imidazo [4,5-c] quinolin-2-yl] methyl] -N-ethylacetamide (compound 112).

A solution of 2 - [(ethylamino) methyl] -7- (7H-pyrazol-3-yl) 1H-imidazo [4,5-c] quinolin-4-amine (75.7 g, 245 , 97 mmol, 1.00 equiv, example 3a) in dichloromethane (2 L). To the solution were added TEA (124 g, 1.23 mol, 5.00 eq.) And acetic anhydride (50 g, 490.20 mmol, 2.00 eq.). The resulting solution was stirred for 6 hours at room temperature. The resulting mixture was then concentrated in vacuo. The residue was dissolved in 3 L of MeOH and allowed to react with stirring for an additional 16 h while the temperature was maintained at 80 ° C. in an oil bath. The reaction mixture was cooled to room temperature and evaporated to give the crude product. The crude material was stirred with 2 L of DCM and the solid was collected by filtration. This procedure was repeated three times. The resulting solid was stirred with 3 L of water and the pH was adjusted to 10 with the addition of aqueous NH ₃ . The precipitated solid was collected by filtration. The mother liquor was evaporated and the resulting precipitate was collected by filtration (3X). The combined solid was dissolved in methanol and stirred with ~ 10% Sithiol at reflux for 4 to 6 hours and filtered. The filtrate was evaporated to give the crude product. This procedure was repeated until the Pd content of the resulting product was <50 ppm. The crude product was washed with 3x250 ml water, collected by filtration and dried. This gave 20.28 g (24%) of compound 112 as a light yellow solid.

¹ H NMR (400 MHz, MeOD, ppm) δ: 8.15-8.07 (m, 1 H), 7.98 (s, 1H), 7.80-7.67 (m, 2 H), 6.80-6.70 (m, 1H), 4.88-4.85 (m, 2H), 3.60-3.48 (m, 2H), 2.30-2.20 (m, 3 H), 1.23 (t, J = 7.2 Hz, 2 H), 1.11 (t, J = 7.2 Hz, 1H); LC / MS [M ++ H] 350.1, LC / MS process conditions: Column: Shim-pack XR-ODS 3.0 x 50 mm, 2.2 μm particles; mobile phase A: water with 0.05% TFA; Mobile Phase B: Acetonitrile with 0.05% TFA; temperature: 40 ° C; gradient: 5% to 100% B over 3.8 minutes; flow rate: 1.2 ml / min; 5 min run. LC RT 1.349 min.

Preparation of N- [[4-amino-7- (1 H-pyrazol-3-yl) -1 H-imidazo [4,5-c] quinolin-2-yl] methyl] -N-ethylacetamide (compound 112, alternative technique).

2 - [(Ethylamino) methyl] -7- (7H-pyrazol-3-yl) -7H-imidazo [4,5-c] quinolin-4-amine hydrochloride (107 g, 311 mmol) and CH ₂ Cl ₂ ( 1.6 L) was stirred at room temperature. Et ₃ N (130 ml, 3 eq.) Was added over 15 to 30 min while maintaining the temperature between 15 and 25 ° C. Acetic anhydride (41 ml, 1.4 eq.) Was added over 15 to 30 minutes and the reaction mixture was stirred for 90 minutes. CH3OH (54 ml) was added and the reaction mixture was stirred for 20 min then concentrated under reduced pressure. The crude material was treated with MeOH (2.2 L) and stirred at 60 to 70 ° C for 2 hours. After cooling to room temperature, the crude product was collected by filtration and washed with CH ₃ OH (220 ml). The resulting solid was dried in vacuo and then stirred in n-butanol (5.4 L) and H2O (1.3 L). The mixture was heated to 60 ° C and the pH was adjusted to 8 with 20% aqueous Na ₂ CO ₃ solution (~ 60 ml). The organic layer was separated and the aqueous layer was extracted with an additional portion of n-butanol (540 ml) at 60 ° C. The combined organic layers were washed with saturated brine (540 ml) at 50 ° C for 15 min (2 times) and concentrated under reduced pressure. The crude product was treated with CH ₃ OH (7.6 L) and H ₂ O (540 ml) and heated to 70 ° C. SiliaMetS-Thiol (22 g) was added and the mixture was stirred for 2 h, then filtered at 60 ° C. The filter cake was washed with CH ₃ OH (220 ml). The filtrate was concentrated under reduced pressure to approx. 800 ml and stirred at room temperature for 2 hours. The precipitate was filtered off and washed with MeOH (220 ml) and H ₂ O (2 x 220 ml). The resulting solid was dried under vacuum and N ₂ , then transferred to a 100 ° C. vacuum oven to complete the drying process. As a result, 83.5 g (77%) of compound 112 were obtained.

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Example 4b. An example of a procedure for the formation of amide

HCI salt

Preparation of N - {[4-amino-7- (1H-pyrazol-3-yl) -1H-imidazo [4,5-c] quinolin-2-yl] methyl} -N-ethyl-3-hydroxy-3- methylbutanamide (compound 440).

To a suspension of 3-hydroxy-3-methylbutanoic acid (10.31 mg, 0.087 mmol) and 2 - ((ethylamino) methyl) 7- (1H-pyrazol-3-yl) -1H-imidazo [4,5-c] quinolin-4-amine, HCl (20 mg, 0.058 mmol) in DMF (582 μl), Hunig's base (25.4 μl, 0.145 mmol) and 2,4,6-tripropyl-1,3,5,2,4 , 6-trioxatriphosphinan 2,4,6-trioxide (50% in DMF) (37.2 μL, 0.064 mmol). The reaction mixture was stirred at room temperature overnight, then diluted with MeOH and purified by preparative HPLC under the following conditions: column: XBridge C18, particle size 200 mm x 19 mm, 5 μm; Mobile phase A: 5:95 acetonitrile: water with 0.1% trifluoroacetic acid; mobile phase B: 95: 5 acetonitrile: water with 0.1% trifluoroacetic acid; gradient: 0 min hold at 13% B, 13 to 38% B for 25 min, then 2 min hold at 100% B; flow rate: 20 ml / min; column temperature: 25 ° C. Fractions containing the desired product were combined and dried by centrifugal evaporation to give N {[4-amino-7- (1H-pyrazol-3-yl) -1H-imidazo [4,5-c] quinolin-2-yl] methyl} -Noethyl-3-hydroxy-3-methylbutanamide as bis-trifluoroacetate salt (8.4 mg, 22%). Ή NMR (500 MHz, DMSO-d ₆ ) δ 8.78 (br C, J = 14.6 Hz, 1H), 8.38-8.12 (m, 2H), 8.00 (br C , 1H), 7.81 (s, 1H), 6.82 (brs, 1H), 5.03-4.73 (m, 2H), 3.653.41 (m, 2H), 2.64- 2.54 (m, 2H), 1.31-0.84 (m, 9H). LC / MS conditions: Column: Waters Acquity UPLC BEN C18, 2.1 x 50 mm particles, 1.7 μm; Mobile Phase A: 5:95 acetonitrile: water with 10 mM ammonium acetate; mobile phase B: 95: 5 acetonitrile: water with 10 mM ammonium acetate; temperature: 50 ° C; gradient: 0 to 100% B over 3 min, then held 0.75 min at 100% B; flow rate: 1.0 ml / min. LC RT: 0.933 min. M / Z = 408.3.

Example 4c. An example of a procedure for the formation of carbamate

HCI salt

Preparation of ethyl N - {[4-amino-7- (1 H-pyrazol-3-yl) -1 H-imidazo [4,5-c] quinolin-2-yl] methyl} -N-ethyl carbamate (compound 436 ).

2 - ((Ethylamino) methyl) -7- (1H-pyrazol-3-yl) -1H-imidazo [4,5-c] quinolin-4-amine (15 mg, 0.049 mmol) was suspended in CH ₂ Cl ₂ ( 488 μl) at room temperature. Pyridine (15.79 μl, 0.195 mmol) was added followed by ethyl chloroformate (9.37 μl, 0.098 mmol). After 2 days, the reaction mixture was concentrated and redissolved in MeOH. TEA (13.60 μl, 0.098 mmol) was added and the reaction mixture was stirred at room temperature overnight. The reaction mixture was diluted with MeOH and purified by preparative LC / MS under the following conditions: column: XBridge C18, 200 mm x 19 mm particles, 5 μm; Mobile Phase A: 5:95 acetonitrile: water with 10 mM ammonium acetate; mobile phase B: 95: 5 acetonitrile: water with 10 mM ammonium acetate; gradient: 0 min hold at 4% B, 4 to 44% B for 20 min, then 4 min hold at 100% B; flow rate: 20 ml / min; column temperature: 25 ° C. Fractions containing the desired product were combined and dried by centrifugal evaporation to give ethyl N - {[4-amino-7- (1H-pyrazol-3-yl) -1Nimidazo [4,5-c] quinolin-2-yl] methyl } -N-ethyl carbamate (7.4 mg, 39%). 1H NMR (500 MHz, DMSO-d ₆ ) δ 8.12 (br C, J = 8.2 Hz, 1H), 7.98 (s, 1H), 7.70 (br C, J = 8 , 5 Hz, 2H), 6.77 (s, 1H), 4.72 (s, 2H), 4.11 (br. C, 2H), 3.52-3.32 (m, 2H), 1 , 29-1.11 (m, 3H), 1.08 (br T, J = 6.9 Hz, 3H). LC / MS conditions: Column: Waters XBridge C18, 2.1 mm x 50 mm particles, 1.7 μm; Mobile Phase A: 5:95 acetonitrile: water with 10 mM ammonium acetate; mobile phase B: 95: 5 acetonitrile: water with 10 mM ammonium acetate; temperature: 50 ° C; gradient: 0% B to 100% B over 3 min, then held at 100% B for 0.75 min; flow rate - 96 037780 ka: 1 ml / min. LC RT: 1.29 min. M / Z = 390.0.

Example 4d. An example of a procedure for the formation of sulfonamide

Preparation of Y - {[4-amino-7- (1H-pyrazol-3-yl) -1H-imidazo [4,5-c] quinolin-2-yl] methyl} -N-ethylpropane-2-sulfonamide (compound 459 ).

To a solution of propan-2-sulfonyl chloride (10.37 mg, 0.073 mmol) in DMF (582 μl) was added 2 ((ethylamino) methyl) -7- (1H-pyrazol-3-yl) -1H-imidazo [4.5 -c] quinolin-4-amine, HCl (20 mg, 0.058 mmol) and Hunig's base (25.4 μl, 0.145 mmol). The reaction mixture was stirred at room temperature overnight, then diluted with DMF and purified by preparative LC / MS under the following conditions: column: XBridge C18, 200 mm x 19 mm particles, 5 μm; Mobile phase A: 5:95 acetonitrile: water with 0.1% trifluoroacetic acid; mobile phase B: 95: 5 acetonitrile: water with 0.1% trifluoroacetic acid; gradient: 0 min hold at 3% B, 3 to 43% B for 30 min, then 4 min hold at 100% B; flow rate: 20 ml / min; column temperature: 25 ° C. Fractions containing the desired product were combined and dried by centrifugal evaporation to give Y - {[4-amino-7- (1H-pyrazol-3-yl) -1H-imidazo [4,5-c] quinolin-2-yl ] methyl} -Y-ethylpropane-2-sulfonamide (1.4 mg, 4.6%). ¹ H NMR (500 MHz, DMSO ^) δ 8.28 (br C, 1H), 8.20 (br C, 1H), 8.02-7.95 (m, 1H), 7.89- 7.82 (m, 1H), 6.83 (br s, 1H), 4.73 (br s, 2H), 3.53-3.40 (m, 2H), 3.31 (br. s, 1H), 1.24 (br s, J = 6.4 Hz, 6H), 1.13 (br t, J = 6.9 Hz, 3H). LC / MS conditions: Column: Waters XBridge C18, 2.1 mm x 50 mm particles, 1.7 μm; Mobile Phase A: 5:95 acetonitrile: water with 10 mM ammonium acetate; mobile phase B: 95: 5 acetonitrile: water with 10 mM ammonium acetate; temperature: 50 ° C; gradient: 0% B to 100% B over 3 min, then held at 100% B for 0.75 min; flow rate: 1 ml / min. LC RT: 1.24 min. M / Z = 413.9.

Example 4f. Example of a Procedure for the Formation of Urea with Carbamoyl Chlorides

Preparation of Y - {[4-amino-7- (1H-pyrazol-3-yl) -1H-imidazo [4,5-c] quinolin-2-yl] methyl} -Y-ethylmorpholine-4-carboxamide (compound 455 ).

To a suspension of 2 - ((ethylamino) methyl) -7- (1H-pyrazol-3-yl) -1H-imidazo [4,5-c] quinolin-4-amine, HCl (20 mg, 0.058 mmol) in DMF ( 582 μl) Hunig's base (25.4 μl, 0.145 mmol) and morpholine-4carbonyl chloride (10.18 μl, 0.087 mmol) were added. After approximately 1.5 h, the reaction was quenched with MeOH, filtered through a syringe filter and purified by preparative HPLC under the following conditions: column: XBridge C18, 200 mm x 19 mm particles, 5 μm; Mobile phase A: 5:95 acetonitrile: water with 10 mM ammonium acetate; mobile phase B: 95: 5 acetonitrile: water with 10 mM ammonium acetate; gradient: 0 min hold at 0% B, 0 to 40% B for 20 min, then 4 min hold at 100% B; flow rate: 20 ml / min; column temperature: 25 ° C. Fractions containing the desired product were combined and dried by centrifugal evaporation to give Y - {[4-amino-7- (1H-pyrazol-3-yl) -1Nimidazo [4,5-c] quinolin-2-yl] methyl} - N-ethylmorpholine-4-carboxamide (14.9 mg, 60%). Ή NMR (500 MHz, DMSO-d ₆ ) δ 8.09 (br C, J = 7.9 Hz, 1H), 8.01-7.90 (m, 1H), 7.74-7.63 (m, 2H), 6.76 (s, 1H) 4.59 (s, 2H), 3.60 (br C, 2H), 3.27-3.18 (m, 4H), 1.90 (s, 4H), 1.10 (br T, J = 6.7 Hz, 3H). LC / MS conditions: Column: Waters Acquity UPLC BEN C18, 2.1 x 50 mm particles, 1.7 μm; Mobile Phase A: 5:95 acetonitrile: water with 10 mM ammonium acetate; mobile phase B: 95: 5 acetonitrile: water with 10 mM ammonium acetate, temperature: 50 ° C; gradient: 0 to 100% B over 3 min, then hold 0.75 min at 100% B; flow rate: 1.0 ml / min. LC RT: 0.914 min, M / Z = 421.4.

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Example 4f. Example of a procedure for the formation of urea with isocyanates

Preparation of 1 - {[4-amino-7- (1H-pyrazol-3-yl) -1H-imidazo [4,5-c] quinolin-2-yl] methyl} -1,3-diethylurea (compound 457).

To a suspension of 2 - ((ethylamino) methyl) -7- (1H-pyrazol-3-yl) -1H-imidazo [4,5-c] quinolin-4-amine, HCl (20 mg, 0.058 mmol) in DMF ( 582 μl) were added Hunig's base (25.4 μl, 0.145 mmol) and isocyanatoethane (5.76 μl, 0.073 mmol). After 1.5 h, the reaction mixture was quenched with MeOH, filtered through a syringe filter and purified by preparative HPLC under the following conditions: column: XBridge C18, 200 mm x 19 mm, 5 μm particles; Mobile Phase A: 5:95 acetonitrile: water with 10 mM ammonium acetate; mobile phase B: 95: 5 acetonitrile: water with 10 mM ammonium acetate; gradient: 0 min hold at 0% B, 0 to 40% B for 20 min, then 4 min hold at 100% B; flow rate: 20 ml / min; column temperature: 25 ° C. Fractions containing the desired product were combined and dried by centrifugal evaporation to give 1 - {[4-amino-7- (1H-pyrazol-3-yl) -1H-imidazo [4,5-c] quinoline-2yl] methyl } -1,3-diethylurea (4.8 mg, 21%). ¹ H NMR (500 MHz, DMSO-d ₆ ) δ 8.13 (br C, J = 8.2 Hz, 1H), 7.97 (s, 1H), 7.73-7.63 (m, 2H), 6.81-6.73 (m, 1H) 6.58 (br C, 1h), 4.68 (s, 2H), 3.36-3.26 (m, 2H), 3, 17-3.07 (m, 2H), 1.09-0.96 (m, 6H). LC / MS conditions: Column: Waters XBridge C18, 2.1 mm x 50 mm particles, 1.7 μm; Mobile Phase A: 5:95 acetonitrile: water with 10 mM ammonium acetate; Mobile phase B: 95: 5 acetonitrile: water with 10 mM ammonium acetate; temperature: 50 ° C; gradient: 0% B to 100% B over 3 min, then held at 100% B for 0.75 min; flow rate: 1 ml / min. LC RT: 0.82 min. M / Z = 379.3.

Example 4g. Example Method for Sulfonylurea Formation

Preparation of 2 - {[(dimethylsulfamoyl) (ethyl) amino] methyl} -7- (1H-pyrazol-3-yl) -1H-imidazo [4,5-c] quinolin-4-amine (compound 458).

To a suspension of 2 - ((ethylamino) methyl) -7- (1H-pyrazol-3-yl) -1H-imidazo [4,5-c] quinolin-4-amine, HCl (20 mg, 0.058 mmol) in DMF ( 582 μl) Hunig's base (25.4 μl, 0.145 mmol) and dimethylsulfamoyl chloride (10.44 mg, 0.073 mmol) were added. The reaction mixture was stirred at room temperature overnight, then quenched with MeOH, filtered through a syringe filter and purified by preparative LC / MS under the following conditions: column: XBridge C18, 200 mm x 19 mm, 5 μm particles; Mobile phase A: 5:95 acetonitrile: water with 0.1% trifluoroacetic acid; mobile phase B: 95: 5 acetonitrile: water with 0.1% trifluoroacetic acid; gradient: 0 min hold at 16% B, 16 to 40% B for 25 min, then 2 min hold at 100% B; flow rate: 20 ml / min; column temperature: 25 ° C. Fractions containing the desired product were combined and dried by centrifugation to give 2- {[(dimethylsulfamoyl) (ethyl) amino] methyl} -7- (1 H-pyrazol-3-yl) -1 H-imidazo [4.5 s] quinolin-4-amine as bis-trifluoroacetate salt (13.5 mg, 35%). ¹ H NMR (500 MHz, DMSO-d ₆ ) δ 8.32 (br C, J = 8.3 Hz, 1H), 8.22 (br C, 1H), 8.00 (br C, J = 7.8 Hz, 1H), 7.82 (br s, 1H), 6.82 (d, J = 1.9 Hz, 1H), 4.68 (s, 2H), 3.43 ( q, J = 7.0 Hz, 2H), 2.75 (s, 6H), 1.17 (t, J = 7.1 Hz, 3H). LC / MS conditions: Column: Waters XBridge C18, 2.1 mm x 50 mm particles, 1.7 μm; Mobile Phase A: 5:95 acetonitrile: water with 10 mM ammonium acetate; mobile phase B: 95: 5 acetonitrile: water with 10 mM ammonium acetate; temperature: 50 ° C; gradient: 0% B to 100% B over 3 min, then held at 100% B for 0.75 min; flow rate: 1 ml / min. LC RT: 1.17 min. M / Z = 415.04.

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Example 4h. An example of an amine formation procedure

Preparation of 2 - {[ethyl (2-methylpropyl) amino] methyl} -7- (1H-pyrazol-3-yl) -1H-imidazo [4,5-c] quinolin-4-amine (compound 453).

To a suspension of isobutyraldehyde (20.97 mg, 0.291 mmol) and 2 - ((ethylamino) methyl) -7- (1H-pyrazol-3yl) -1H-imidazo [4,5-c] quinolin-4-amine, HCl ( 20 mg, 0.058 mmol) in MeOH (582 μl) was added sodium triacetoxyborohydride (37.0 mg, 0.175 mmol). After 4.5 hours, isobutyraldehyde (20.97 mg, 0.291 mmol) and sodium triacetoxyborohydride (37.0 mg, 0.175 mmol) were added. After 2.75 h, the reaction mixture was partially concentrated, diluted with water and extracted three times with EtOAC. The organic layers were concentrated. The residue was dissolved in DMF, filtered through a syringe filter and purified by preparative HPLC under the following conditions: column: XBridge C18, 200 mm x 19 mm, 5 μm particles; Mobile Phase A: 5:95 acetonitrile: water with 10 mM ammonium acetate; mobile phase B: 95: 5 acetonitrile: water with 10 mM ammonium acetate; gradient: 0 min hold at 20% B, 20 to 44% B for 25 min, then 2 min hold at 100% B; flow rate: 20 ml / min; column temperature: 25 ° C. Fractions containing the desired product were combined and dried by centrifugal evaporation to give 2 {[ethyl (2-methylpropyl) amino] methyl} -7- (1H-pyrazol-3-yl) -1H-imidazo [4,5-c] quinoline -4-amine (1.1 mg, 5.2%). 1H NMR (500 MHz, DMSO-de) δ 8.15 (d, J = 8.2 Hz, 1H), 7.98 (s, 1H), 7.72-7.64 (m, 2H), 6 , 73 (d, J = 1.9 Hz, 1H), 3.86 (s, 2H), 2.61 (q, J = 7.0 Hz, 2H), 2.27 (d, J = 7, 1 Hz, 2h), 1.78-1.69 (m, 1H), 1.06 (t, J = 7.0). Hz, 3H), 0.86 (d, J = 6.6 Hz, 6H). LC / MS conditions: Column: Waters XBridge C18, 2.1 mm x 50 mm particles, 1.7 μm; Mobile Phase A: 5:95 acetonitrile: water with 10 mM ammonium acetate; mobile phase B: 95: 5 acetonitrile: water with 10 mM ammonium acetate; temperature: 50 ° C; gradient: 0% B to 100% B over 3 min, then held at 100% B for 0.75 min; flow rate: 1 ml / min; LC RT: 1.36 min; M / Z = 364.1.

Table connections 4 were obtained using the procedures described above. LC / MS conditions: column: VEN C18 2.1 x 50 mm; mobile phase A: water with 0.05% TFA; Mobile Phase B: Acetonitrile with 0.05% TFA; temperature: 50 ° C; gradient: 2-98% B over 1.7 minutes; flow rate: 0.8 ml / min.

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Table 4

Connection NAME LC / MS [M ⁺ + H] LCRT NMR data 102 N - ((4-Amhho-7- (1H- pyrazol-1-yl) -1 Imidazo [4,5c] quinolin-2yl) methyl) -M- methylacetamide 336.3 0.54 min Ή NMR (400 MHz, METHANOL d ₄ ) δ 8.36 -8.31 (m, 1H), 8.268.16 (m, 1H), 8.02-7.96 (m, 1H), 7.81 -7 , 75 (m, 2H), 6.61 -6.58 (m, 1H), 4.95-4.92 (m, 2H), 3.23 (s, 2H), 3.07 (s, 1H ), 2.32 (s, 1H), 2.26 (s, 2H) 106 2 - ((Ethylamino) methyl) 1-methyl-7- (1Hpyrazol-3-yl) -1Nimidazo [4,5c] quinolin-4-amine 322.1 0.48 min Ή NMR (400 MHz, METHANOL d ₄ ) δ 8.30 (bRd, 7 = 8.6 Hz, 1H), 8.15-7.98 (m, 1H), 7.89 -7.64 (m, 2H ), 6.79 (s, 1H), 4.24 (s, 3H), 4.12 (s, 2H), 2.80 (q, 7 = 7.2 Hz, 2H), 1.21 (t , 7 = 7.2 Hz, 3H) 108 N - ((4-Amhho-7- (1H- pyrazol-1-yl) -1Nimidazo [4,5с] quinolin-2yl) methyl) -Methylmethanesulfonamide 386.3 0.59 min Ή NMR (400 MHz, METHANOL d ₄ ) δ 8.34 (d, 7 = 2.4 Hz, 1H), 8.30 -8.20 (m, 1H), 7.99 (d, 7 = 2.1 Hz, 1H), 7.82-7.75 (m, 2H), 6.59 (t, 7 = 2.1 Hz, 1H), 4.77 (s, 2H), 3.46 (q, 7 = 7.1 Hz, 2H), 3.04 (s, 3H), 1.22 (t, 7 = 7.1 Hz, 3H) 109 tert-Butyl - ((4-amino7- (1H-pyrazol-3-yl) 1H-imidazo [4,5с] quinolin-2yl) methyl) (ethyl) carbam at 408.3 0.69 min Ή NMR (400 MHz, METHANOL d ₄ ) δ 8.22-8.09 (m, 1H), 8.07 7.98 (m, 1H), 7.91-7.51 (m, 2H), 6, 79 (bRs, 1H), 4.77 (s, 2H), 3.63 -3.42 (m, 2H), 1.64-1.32 (m, 9H), 1.19 (bRs, 3H)

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110 1 - ((4-Amino-7- (1H- pyrazol-3-yl) -1 imidazo [4,5c] quinolin-2yl) methyl) -1-ethyl-3,3 dimethylurea 379.3 0.58 min Ή NMR (400 MHz, METHANOL d ₄ ) δ 8.25-8.10 (m, 1H), 8.097.99 (m, 1H), 7.91 -7.65 (m, 2H), 6.79 (bRs , 1H), 4.67 (s, 2H), 3.39 -3.34 (m, 2H), 2.97 (s, 6H), 1.23 (t, 7 = 7.1 Hz, 3H) of N - ((4-Amhho-7- (1H- pyrazol-3-yl) -1Nimidazo [4,5с] quinolin-2yl) methyl) acetamide 322.3 0.49 min Ή NMR (400 MHz, METHANOL d ₄ ) δ 8.13 (bRd, 7 = 8.3 Hz, 1H), 8.03 (bRs, 1H), 7.88 -7.65 (m, 2H), 6, 78 (s, 1H), 4.71 (s, 2H), 2.11 (s, 3H) 158 N - ((4-Amhho-7methoxy-lHimidazo [4,5c] quinoline-2yl) methyl) -ethylacetamide 314.3 0.55 min Ή NMR (400 MHz, METHANOL d ₄ ) δ 8.07 -7.95 (m, 1H), 7.13 (d, 7 = 2.2 Hz, 1H), 7.07 -6.97 (m, 1H ), 3.92 (s, 1H), 3.91 (s, 2H), 3.61 -3.50 (m, 2H), 2.29 (s, 1H), 2.26 (s, 2H) , 1.26 (t, 7 = 7.1 Hz, 2H), 1.13 (t, 7 = 7.1 Hz, 1H) 419 N - ((4-Amhho-7 (thiophen-3-yl) -1Nimidazo [4,5c] quinolin-2yl) methyl) -ethylacetamide 366.3 0.65 min Ή NMR (400 MHz, METHANOL d ₄ ) δ 8.20 -8.10 (m, 1H), 7.947.90 (m, 1H), 7.80 -7.75 (m, 1H), 7.75-7 , 67 (m, 1H), 7.62-7.53 (m, 2H), 4.91 (s, 2H), 3.66 -3.52 (m, 2H), 2.31 (s, 1H ), 2.27 (s, 2H), 1.27 (t, 7 = 7.2 Hz, 2H), 1.15 (t, 7 = 7.1 Hz, 1H)

Table connections 5 were obtained using the procedures described above.

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Table 5

Connection NAME LC / MS [M ⁺ + H] LCRT LC Way 436 ethyl N-J [4-amino-7- (1 H-pyrazol-3yl) -1 H-imidazo [4,5-c] quinoline-2yl] methyl} -N-ethyl carbamate 390.0 1.29 min BUT 437 N- {[4-amino-7- (1 H-pyrazol-3-yl) -1 Nimid azo [4,5-c] quinolin-2-yl] methyl} X-ethyl-2- (3-methylphenyl) acetamide 440.4 1.30 min BUT 438 N- {[4-amino-7- (1 H-pyrazol-3-yl) -1 Nimid azo [4,5-c] quinolin-2-yl] methyl} N-ethylpyridine-3-carboxamide 413.0 1DZ min IN 439 N- {[4-amino-7- (1 H-pyrazol-3-yl) -1 Nimid azo [4,5-s] quinolin-2-yl] methyl} M-ethylpyridine-2-carboxamide 413.2 1.19 min IN 440 N- {[4-amino-7- (1 H-pyrazol-3-yl) -1Nimide azo [4,5-c] quinolin-2-yl] methyl} - N-ethyl-3-hydroxy-3-methylbutanamide 408.4 0.97 min IN 441 N- {[4-amino-7- (1 H-pyrazol-3-yl) -1 Nimid azo [4,5-c] quinolin-2-yl] methyl} H-ethyl-4-methoxybenzamide 442.01 1.21 min BUT 442 N- {[4-amino-7- (1 H-pyrazol-3-yl) -1Nimid azo [4,5-c] quinolin-2-yl] methyl} Nethylpyrazine-2-carboxamide 414.0 0.88 min BUT 443 N- {[4-amino-7- (1 H-pyrazol-3-yl) -1 Nimid azo [4,5 -c] quinolin-2-yl] methyl} N-ethyl-2-φ enyl acetamide 426.0 1.26 min IN 444 N- {[4-amino-7- (1 H-pyrazol-3-yl) -1 Nimide azo [4,5-c] quinolin-2-yl] methyl} N-ethyl-4-φ enyl butanamide 454.1 1.48 min BUT 445 N- {[4-amino-7- (1 H-pyrazol-3-yl) -1 Nimide azo [4,5-c] quinolin-2-yl] methyl} H-ethyl-4-methylpentanamide 406.1 1.41 min BUT 446 N- {[4-amino-7- (1 H-pyrazol-3-yl) -1 Nimid azo [4,5-c] quinolin-2-yl] methyl} N-ethyl-1-methyl-1 H-pyrrole -2carboxamide 415.2 1.17 min BUT 447 N- {[4-amino-7- (1 H-pyrazol-3-yl) -1 Nimid azo [4,5-c] quinolin-2-yl] methyl} N-ethyl-3-phenylpropanamide 440.2 1.37 min BUT

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448 Ν- {[4-amino-7- (1 H-pyrazol-3-yl) -1 Nimid azo [4,5 -c] quinolin-2-yl] methyl} N-ethyl-1,3-thiazole-4- carboxamide 419.3 1.12 min BUT 449 Ν- {[4-amino-7- (1 H-pyrazol-3-yl) -1 Nimid azo [4,5-c] quinolin-2-yl] methyl} 2-chloro-M-ethylbenzamide 446.0 1.28 min BUT 450 Ν- {[4-amino-7- (1 H-pyrazol-3 -yl) -1Nimid azo [4,5-s] quinolin-2-yl] methyl} H-ethylpyridine-4-carboxamide 413.0 0.92 min BUT 451 Ν- {[4-amino-7- (1 H-pyrazol-3-yl) -1 Nimid azo [4,5-c] quinolin-2-yl] methyl} N-ethyl-1-methyl-1 H-ind ol-3 carboxamide 465.2 1.31 min IN 452 Ν- {[4-amino-7- (1 H-pyrazol-3-yl) -1Nimid azo [4,5 -c] quinolin-2-yl] methyl} 4-chloro-M-ethylbenzamide 446.0 1.36 min BUT 453 2 - {[ethyl (2methylpropyl) amino] methyl} -7- (1Hpyrazol-3-yl) -1 H-imidazo [4.5 s] quinolin-4-amine 364.1 1.36 min BUT 454 1 - {[4-amino-7- (1H-pyrazol-3-yl) -1Nimide azo [4,5 -c] quinolin-2-yl] methyl} 1,3,3-triethylurea 407.1 1.31 min BUT 455 Ν- {[4-amino-7- (1 H-pyrazol-3-yl) -1Nimid azo [4,5-c] quinolin-2-yl] methyl} H-ethylmorpholine-4-carboxamide 421.4 0.91 min IN 456 1 - {[4-amino-7- (1 H-pyrazol-3-yl) -1 Nimid azo [4,5 -c] quinolin-2-yl] methyl} 3 -tert-but yl-1-ethylurea 407.1 1.29 min BUT 457 1 - {[4-amino-7- (1H-pyrazol-3-yl) -1Nimide azo [4,5 -c] quinolin-2-yl] methyl} 1,3-diethylurea 379.3 0.82 min BUT 458 2- {[(dimethylsulfamoyl) (ethyl) amino] 415.0 1.17 min BUT

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methyl} -7- (1H-pyrazol-3-yl) -1Nimidazo [4,5 -c] quinolin-4-amine 459 N- {[4-amino-7- (1 H-pyrazol-3-yl) -1 Nimid azo [4,5-c] quinolin-2-yl] methyl} N-ethylpropane-2-sulfonamide 413.9 1.24 min BUT 460 N- {[4-amino-7- (1 H-pyrazol-3-yl) -1 Nimid azo [4,5-c] quinolin-2-yl] methyl} N-ethylpropane-1 -sulfonamide 414.0 1D6 min BUT 470 N- {[4-amino-1 - (2-hydroxyethyl) -7 (thiophen-2-yl) -1H-imidazo [4,5c] quinolin-2-yl] methyl} -Nethylpropanamide 424.1 1.24 min D 472 N- {[4-amino-1-ethyl-7- (thiophen-2-yl) 1H-imidazo [4,5-c] quinolin-2yl] methyl} -N-ethyl acetamide 393.9 2.27min E 473 N- {[4-amino-1-ethyl-7- (1 H-pyrazol-5 yl) -1 H-imidazo [4,5-c] quinolin-2yl] methyl} -N-ethyl propanamide 392.3 1.75 min E 474 N- {[4-amino-1-ethyl-7- (1 H-pyrazol-5 yl) -1 H-imidazo [4,5-c] quinolin-2yl] methyl} -N-ethyl acetamide 378.3 1.95 min FROM 475 N- {[4-amino-1 - (2-hydroxyethyl) -7 (1H-pyrazol-5-yl) - 1H-imidazo [4,5c] quinolin-2-yl] methyl} -Nethylpropanamide 408.1 0.92 min D 476 N- {[4-amino-1 - (2-hydroxyethyl) -7 (1H-pyrazol-1-yl) -1H-imidazo [4,5c] quinolin-2-yl] methyl} -Nethylacetamide 394.1 0.93 min D 478 N- {[4-amino-1 - (2-hydroxyethyl) -7 (1 H-pyrazol-1-yl) -1H-imidazo [4,5c] quinolin-2-yl] methyl} -Nethylpropanamide 408.1 1.00 min D 479 N- {[4-amino-1-ethyl-7- (1 H-pyrazol-1 yl) -1 H-imidazo [4,5-c] quinolin-2yl] methyl} -N-ethyl acetamide 378.0 1.12 min E 480 N- {[4-amino-1-ethyl-7- (thiophen-2-yl) 1H-imidazo [4,5-c] quinolin-2yl] methyl} -N-ethyl propanamide 408.0 1.50 min E 481 N- {[4-amino-1-ethyl-7- (1 H-pyrazol-1 yl) -1 H-imidazo [4,5-c] quinolin-2yl] methyl} -N-ethyl propanamide 392.2 1.33 min FROM 482 N- {[4-amino-1 - (2-hydroxyethyl) -7 (thiophen-2-yl) -1H-imidazo [4,5c] quinolin-2-yl] methyl} -Nethylacetamide 410.1 1.79 min D

LC methods for table. 4: A: Column: Waters XBridge C18, 2.1 mm x 50 mm particles, 1.7 μm; Mobile Phase A: 5:95 acetonitrile: water with 10 mM ammonium acetate; mobile phase B: 95: 5 acetonitrile:

- 104 037780 water with 10 mM ammonium acetate; temperature: 50 ° C; gradient: 0% B to 100% B over 3 min, then held at 100% B for 0.75 min; flow rate: 1 ml / min. B: Column: Waters Acquity UPLC BEN C18, 2.1 x 50 mm, 1.7 μm particles; Mobile Phase A: 5:95 acetonitrile: water with 10 mM ammonium acetate; mobile phase B: 95: 5 acetonitrile: water with 10 mM ammonium acetate, temperature: 50 ° C; gradient: 0 to 100% B over 3 min, then held 0.75 min at 100% B; flow rate: 1.0 ml / min. C: Column: Shim-pack XR-ODS 3.0 mm x 50 mm, 2.2 μm particles; mobile phase A: water with 0.05% TFA; Mobile Phase B: Acetonitrile with 0.05% TFA; temperature: 40 ° C; gradient: 0% B to 95% B over 2 min, then held at 95% B for 0.7 min; flow rate: 1.5 ml / min. D: Column: Kinetex EVO, 3.0 mm x 50 mm, 2.6 μm particles; mobile phase A: water with 0.03% NH ₃ H ₂ O; mobile phase B: acetonitrile; temperature: 40 ° C; gradient: 10% B to 95% B over 2 min, then held at 95% B for 0.60 min; flow rate: 1.2 ml / min. E: Column: XBridge VEN Shield RP18, 2.1 mm x 50 mm, 2.5 μm particles; mobile phase A: water with 6.5 mM ammonium bicarbonate; mobile phase B: acetonitrile; temperature: 40 ° C; gradient: 10% B to 50% B over 2.2 minutes, then to 95% B over 0.60 minutes, then held at 95% B for 0.70 minutes; flow rate: 1 ml / min.

Example 5. An example of a method for producing analogs, where R1 / R ² is lactam and R ⁵ is aryl.

Scheme 8 o

Stage 1. Obtaining 1 - ((7-bromo-1- (4-methoxybenzyl) -1H-imidazo [4,5-c] quinolin-2-yl) methyl) pyrrolidin-2-one

Not selected

To a solution of 7-bromo-N ⁴ - (4-methoxybenzyl) quinoline-3,4-diamine (1 g, 2.8 mmol) and (2-oxopyrrolidin-1-yl) acetic acid (0.450 g, 3.15) mmol)) in DMF (10 ml), HATU (1.4 g, 3.68 mmol) and NEt ₃ (0.78 ml, 5.6 mmol) were added. The mixture was stirred at room temperature for 2 hours and then heated in an oil bath at 60 ° C for 24 hours. The cooled reaction mixture was concentrated to remove all volatiles. Water (30 ml) was added, the precipitate was filtered and dried to give crude 1 - ((7-bromo-1- (4-methoxybenzyl) -1H-imidazo [4,5-c] quinolin-2-yl) methyl) pyrrolidine- 2-one, which was used in the next step without further purification. (1.4 g,> 100% yield) (ES, m / z): [M + H] + = 465.2 / 467.1.

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Stage 2. Obtaining 7-bromo-1- (4-methoxybenzyl) -2 - ((2-oxopyrrolidin-1-yl) methyl) -1Nimidazo [4,5-c] quinoline 5-oxide

To a solution of 1 - ((7-bromo-1- (4-methoxybenzyl) -1H-imidazo [4,5-c] quinolin-2-yl) methyl) pyrrolidin-2one (370 mg, 1.01 mmol) in CHCl ₃ (10 ml) was added m-chloroperoxybenzoic acid (content 70%, 263 mg, 1.5 mmol). The mixture was stirred for 60 h for 3 h, after which the cooled mixture was diluted with saturated aqueous NaHCO ₃ solution and extracted with CH ₂ Cl ₂ (2 x 20 ml). The combined organic layers were washed with brine, dried over Na ₂ SO ₄ , filtered and concentrated to give the crude N-oxide (400 mg, 1.05 mmol, 99% yield) as a brownish foam. (ES, m / z): [M + H] + = 481.2 / 483.1.

Stage 3. Obtaining 1 - ((4-amino-7-bromo-1- (4-methoxybenzyl) -1H-imidazo [4,5-c] quinolin-2-yl) methyl) pyrrolidin-2-one

To a solution of 7-bromo-1- (4-methoxybenzyl) -2 - ((2-oxopyrrolidin-1-yl) methyl) -1H-imidazo [4,5-c] quinoline 5-oxide (400 mg, 1.05 mmol) and NH4OH (10 ml) in dichloromethane (20 ml) cooled in an ice bath, p-toluenesulfonyl chloride (300 mg, 1.57 mmol) in CH ₂ Cl ₂ (10 ml) was added dropwise. The resulting solution was stirred for an additional 30 minutes after the addition was complete. Water (20 ml) was added and the layers were separated. The aqueous layer was once again extracted with CH ₂ Cl ₂ (30 ml). The solution was filtered through a pad of Na ₂ SO ₄ and the filtrate was concentrated in vacuo. The residue was triturated with EtOAc / hexanes (1/3) and dried under high vacuum to give 1 - ((4-amino-7-bromo-1- (4-methoxybenzyl) -1H-imidazo [4,5-c] quinoline- 2-yl) methyl) pyrrolidin-2-one as a yellow solid (317 mg, 0.66 mmol, 66% yield). (ES, m / z): [M + H] + = 480.3 / 482.2.

Stage 4. Obtaining 1 - ((4-amino-1- (4-methoxybenzyl) -7-aryl (heteroaryl) -1H-imidazo [4,5-c] quinolin-2-yl) methyl) pyrrolidin-2-one

To dioxane solution 1 - ((4-amino-7-bromo-1- (4-methoxybenzyl) -1H-imidazo [4,5-c] quinolin-2yl) methyl) pyrrolidin-2-one (400 mg, 833 μmol ) aryl (heteroaryl) boric acid (or aryl boronate ester) (1.5 equiv.), Pd (dppf) Cl ₂ CH ₂ Cl ₂ (50 mg) and an aqueous solution of K2CO3 (10 ml, 2M) were added sequentially. The mixture was irradiated in a Biotage Initiator microwave apparatus at 120 ° C for 10 min. The organic layer was diluted with EtOAc and separated, and the aqueous layer was washed with EtOAc. The combined organic layers were filtered, evaporated and flash chromatographed on silica gel eluting with chloroform / methanol to give pure 1 - ((4-amino-1- (4-methoxybenzyl) -7-aryl (heteroaryl) -1H-imidazo [ 4,5-c] quinolin-2-yl) methyl) pyrrolidin-2-one.

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Stage 5. General procedure for removing protection

The product from the previous step was dissolved in TFA (20 ml) and stirred at 70 ° C for 1 h, after which LCMS analysis showed that the protecting group of PMB was completely cleaved. The mixture was evaporated and flash chromatographed on silica gel eluting with chloroform / methanol to give the product as the free base. The material was dissolved in methanol solution of hydrogen chloride (1N), the solvent was evaporated, and the resulting solid was washed with diethyl ether and dried to give 1 - ((4-amino-7-aryl (heteroaryl) -1H-imidazo [4,5-c ] -quinolin-2-yl) methyl) pyrrolidin-2-one as the hydrochloride salt.

The compounds shown in table. 6 were obtained in accordance with the synthesis methods described above.

Table 6

Compound NAME [M + H] + 279 1 - [(4-amino-7-pyridin-3-yl-1H-imidazo [4,5c] quinolin-2-yl) methyl] pyrrolidin-2-one 359.1 280 1 - [(4-amino-7-phenyl-1 H-imidazo [4,5-c] quinolin-2yl) methyl] pyrrolidin-2-one 358.2 281 1 - [[4-amino-7- (2-chlorophenyl) -1H-imidazo [4,5c] quinolin-2-yl] methyl] pyrrolide in-2-one 392.1 282 1 - [[4-amino-7- (2-methylphenyl) -1 H-imidazo [4,5c] quinolin-2-yl] methyl] pyrrolide in-2-one 372.2 283 1 - [[4-amino-7- (2-methoxyphenyl) -1 H-imide azo [4.5 s] quinolin-2-yl] methyl] pyrrolide in-2-one 388.2 284 1 - [[4-amino-7- (2-fluorophenyl) -1H-imidazo [4,5c] quinolin-2-yl] methyl] pyrrolide in-2-one 376.1 285 1 - [[4-amino-7- (3-chlorophenyl) -1 H-imide azo [4.5 s] quinolin-2-yl] methyl] pyrrolide in-2-one 392.1 286 1 - [[4-amino-7- (3-methylphenyl) -1 H-imidazo [4,5c] quinolin-2-yl] methyl] pyrrolide in-2-one 372.2 287 1 - [[4-amino-7- (3-methoxyphenyl) -1 H-imide azo [4.5 s] quinolin-2-yl] methyl] pyrrolide in-2-one 388.2 288 3 - [4-amino-2- [(2-oxopyrrolide in-1-yl) methyl] -1Nimidazo [4,5 -c] quinolin-7-yl] benzonitrile 383.1 289 1 - [[4-amino-7- (3-fluorophenyl) -1 H-imide azo [4.5 s] quinolin-2-yl] methyl] pyrrolide in-2-one 376.1 291 1 - [[4-amino-7- (4-methylphenyl) -1 H-imidazo [4,5c] quinolin-2-yl] methyl] pyrrolide in-2-one 372.2 292 1 - [[4-amino-7- (4-methoxyphenyl) -1 H-imide azo [4.5 s] quinolin-2-yl] methyl] pyrrolide in-2-one 388.2

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294 1 - [(4-amino-7-bromo-1 H-imide azo [4,5-c] quinolin-2yl) methyl] pyrrolidin-2-one 360.0 295 1 - [(4-amino-7-pyrazol-1-yl-1 H-imide azo [4.5 s] quinolin-2-yl) methyl] pyrrolidin-2-one 348.1 296 1 - [[4-amino-7- (4-methylpyrazol-1-yl) -1 H-imidazo [4.5 s] quinolin-2-yl] methyl] pyrrolide in-2-one 362.2 298 1 - [[4-amino-7- (ohap-4-yl) -1 H-imide azo [4,5-c] quinolin2-yl] methyl] pyrrolidin-2-one 366.2 301 1 - [[4-amino-7- (3-methylpyrazol-1-yl) -1 H-imidazo [4.5 s] quinolin-2-yl] methyl] pyrrolide in-2-one 362.2 302 1 - [(4-amino-7-cyclohexyl-1 H-imid azo [4.5 s] quinolin-2-yl) methyl] pyrrolidin-2-one 364.2 303 1 - [1 - (4-amino-7-bromo-1 H-imidazo [4,5-c] quinolin-2yl) ethyl] pyrrolide IN-2-OH 374.1 304 1 - [[4-amino-7- (6-methoxypyridin-3-yl) -1Nimidazo [4,5-c] quinolin-2-yl] methyl] pyrrolidin-2-one 389.2 307 1 - [[4-amino-7- (2-fluoropyr-idin-3-yl) -1 H-imidazo [4.5 s] quinolin-2-yl] methyl] pyrrolide in-2-one 377.1 308 1 - [(4-amino-7-thiophen-3-yl-1 H-imidazo [4.5 s] quinolin-2-yl) methyl] pyrrolidin-2-one 364.1 309 1 - [(4-amino-7-thiophen-2-yl-1 H-imidazo [4.5 s] quinolin-2-yl) methyl] pyrrolidin-2-one 364.1 315 2- [4-amino-2- [(2-oxopyrrolide in-1-yl) methyl] -1Nimidazo [4,5 -c] quinolin-7-yl] benzonitrile 383.2 316 1 - [(4-amino-7-bromo-1 H-imide azo [4,5-c] quinolin-2yl) methyl] -4-phenylpyrrolidin-2-one 436.1 319 1 - [[4-amino-7- (2-methoxypyridin-3-yl) -1Nimidazo [4,5 -c] quinolin-2-yl] methyl] pyrrolidin-2-one 389.2 320 1 - [[4-amino-7- (2-chloropyr-idin-3-yl) -1 H-imidazo [4.5 s] quinolin-2-yl] methyl] pyrrolide in-2-one 393.1 321 1 - [[4-amino-7- (4-chloropyr-idin-3-yl) -1 H-imidazo [4.5 s] quinolin-2-yl] methyl] pyrrolide in-2-one 393.1 324 1 - [[4-amino-7- [6- (trifluoromethyl) pyridin-3-yl] -1Nimidazo [4,5-c] quinolin-2-yl] methyl] pyrrolidin-2-one 427.1 326 5 - [4-amino-2- [(2-oxopyrrolide in-1-yl) methyl] -1 Nimide azo [4,5-c] quinolin-7-yl] pyridine-3-carbonitrile 384.1 327 5 - [4-amino-2- [(2-oxopyrrolide in-1-yl) methyl] -1Nimide azo [4,5 -c] quinolin-7-yl] pyrid and ine-2-carbonitrile 384.1 328 1 - [[4-amino-7- (5-chloropyr-idin-3-yl) -1 H-imidazo [4.5 s] quinolin-2-yl] methyl] pyrrolide in-2-one 393.1 330 1 - [[4-amino-7- (6-fluoropyridin-3-yl) -1 H-imidazo [4.5 s] quinolin-2-yl] methyl] pyrrolide in-2-one 377.1 331 1 - [[4-amino-7- (5-fluoropyridin-3-yl) -1 H-imidazo [4.5 s] quinolin-2-yl] methyl] pyrrolide in-2-one 377.1 332 1 - [(4-amino-7-cycloheptyl-1H-imidazo [4,5c] quinolin-2-yl) methyl] pyrrolidin-2-one 378.2 333 1 - [(4-amino-7-pyridin-3-yl-1H-imidazo [4,5c] quinolin-2-yl) methyl] -4-phenylpyrrolidin-2-one 435.2

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The compounds shown in table. 7 were obtained in accordance with the synthesis methods described above.

Table 7

Compound NAME LC / MS [M ⁺ + H] LCRT LC Way 463 1 - {[4-amino-1-ethyl-7- (thiophen-2yl) -1 H-imidazo [4,5-c] quinolin-2yl] methyl} pyrrolidin-2-one 392.2 1.16 minutes FROM 465 1 - {[4-amino-1-ethyl-7- (1 H-pyrazol1-yl) -1 H-imide azo [4,5-c] quinolin2-yl] methyl} pyrrolide in-2-one 376.2 0.98 min FROM 466 1 - {[4-amino-1-ethyl-7- (1 H-pyrazol5-yl) -1 H-imide azo [4,5-c] quinolin2-yl] methyl} pyrrolide in-2-one 376.2 0.95 minutes FROM 469 1 - {[4-amino-1 - (2-hydroxyethyl) -7 (thiophen-2-yl) -1H-imidazo [4,5c] quinolin-2yl] methyl} pyrrolidin-2-one 408.0 1.16 minutes BUT

LC Methods: A: Column: Kinetex EVO, 3.0 mm x 50 mm, 2.6 μm particles; mobile phase A: water with 5 mM ammonium bicarbonate; mobile phase B: acetonitrile; temperature: 40 ° C; gradient: 10% B to 95% B over 2 min, then held at 95% B for 0.79 min; flow rate: 1 ml / min; B: Column: Kinetex EVO, 3.0 mm x 50 mm, 2.6 μm; mobile phase A: water with 0.03% NH ₃ H ₂ O; mobile phase B: acetonitrile; temperature: 40 ° C; gradient: 10% B to 95% B over 2 min, then held at 95% B for 0.60 min; flow rate: 1.2 ml / min; C: Column: Express C18 2.1 mm x 50 mm, 2.7 μm particles; mobile phase A: water with 0.05% TFA; Mobile Phase B: Acetonitrile with 0.05% TFA; temperature: 40 ° C; gradient: 5% B to 100% B over 2 min, then held at 100% B for 0.75 min; flow rate: 0.8 ml / min.

Example 6. Biological analyzes.

Measurement of IL-1 production in PMA-differentiated THP-1 cells.

THP-1 cells were purchased from the American Type Culture Collection and subcultured according to the supplier's instructions. Before experiments, cells were cultured in RPMI 1640 medium containing 10% heat-inactivated FBS, penicillin (100 U / ml) and streptomycin (100 μg / ml), and maintained in logarithmic growth phase until experiments. Before the experiment, THP-1 was treated with PMA (phorbol 12-myristate 13-acetate) (10 μg / ml) for 24 hours. buffered saline), centrifuged, resuspended in 2% heat-inactivated FBS with RPMI at a concentration of 1x10 ⁶ cells / ml, and 100 μl was seeded in a 96-well plate. Compounds were dissolved in dimethyl sulfoxide (DMSO) and added to the culture medium to achieve the desired concentration (eg, 100, 30, 10, 3, 1, 0.3, or 0.1 μM). Cells were incubated with compounds for 4 hours. The cell-free supernatant was collected and IL-1e production was assessed by ELISA. A control containing only the vehicle was carried out simultaneously with each experiment. The final concentration of DMSO was 1%. The compounds showed a dose-dependent increase in IL-1e in PMA-differentiated THP-1 cells.

Measurement of IL-1e production in PMA-differentiated THP-1 cells (alternative procedure).

THP-1 cells were purchased from the American Type Culture Collection and subcultured according to the supplier's instructions. Before the experiments, the cells were cultured in RPMI 1640 medium containing 10% heat-inactivated FBS, penicillin (100 U / ml), streptomycin (100 μg / ml), HEPES (10 mM), and sodium pyruvate (1 mM), and kept in logarithmic growth phase prior to experimentation. Before the experiment, THP-1 cells were treated with PMA (phorbol12-myristate-13-acetate) (20 μg / ml) overnight. On the day of the experiment, the medium was removed and the attached cells were treated with trypsin for 2 min, then the cells were harvested, washed with PBS (phosphate buffered saline), pelleted by centrifugation and resuspended in 2% heat-inactivated FBS with RPMI at a concentration of 50,000 cells / well in a 384-well tablet. Demon

109 037780 cell supernatant was harvested and IL-Ιβ production was assessed by ELISA. Compounds were dissolved in dimethyl sulfoxide (DMSO) and added to the culture medium to achieve the desired concentration (eg, 100, 30, 10, 3, 1, 0.3, or 0.1 μM). Cells were incubated with compounds for 2 hours. A control containing only vehicle was run concurrently with each experiment. The final concentration of DMSO was 1%. The compounds showed a dose-dependent increase in IL-1 β in differentiated PMA THP-1 cells.

Measurement of IL-1e production - hTRF protocol (second alternative procedure).

Serial dilutions of compounds in DMSO were added to small volume 384-well plates at 100 nl / well using an ECHO 550 acoustic pipette (Labcyte) to achieve a final initial concentration of 10 μM in the assay.

THP-1 cells in RPMI medium (Gibco, 11875) with 10% FBS at a density of 1 × 10 ⁶ cells / ml in a T175 flask were treated with phorbol-12-myristate-13-acetate (PMA) (Sigma, P1585) with a final concentration of 50 ng / ml, overnight at 37 ° C in 5% CO ₂ for differentiation. Cells were harvested the next day after thorough washing with dPBS using 0.5% trypsin. The cell solution was prepared from 1x10 ⁶ cells / ml at 50,000 cells in 50 μl / well in RPMI medium with 2% FBS. Cells were plated using a multichannel pipette on compound dilutions in 384-well, black transparent bottom, tissue culture-treated Greiner plates (781090). The plates were incubated in an incubator at 37 ° C at 5% CO2 for 2 h.

After a 2-hour incubation, the cell plates were centrifuged for 5 min at 1200 rpm. Using Felix (CyBio), 8 μl of supernatant was transferred to 384 well, low volume, white proxy plates. (Perkin Elmer, 6008230). The hTRF Human IL-1e Detection Kit (CISBIO, 62HIL1BPEG) was used for the analysis of the supernatant. The kit instructions were followed to obtain the ILlBeta standard curve, and then the antibodies from the kit were diluted 1:40 rather than 1:20 as indicated in the kit instructions. After combining, these antibodies were added to all plates, 5 μl / well. The plates were covered and incubated at 4 ° C overnight. Plates were then read on a Perkin Elmer EnVision at 665/615 nm using an hTRF laser. The compounds showed a dose-dependent increase in the increase in IL-1e.

Measurement of IL-1e Production - Human Whole Blood Assay.

Serial dilutions of compounds in DMSO were added to 384-well low volume plates at 100 nl / well using an ECHO 550 acoustic dispenser (Labcyte) to achieve a final initial concentration of 10 μM in the assay. Human venous whole blood from healthy donors was pretreated with LPS (Invivogen, Cat # tlrl-eblps) at 1 ng / ml for four hours at 37 ° C. in a humid air incubator at 95% air / 5% CO ₂ . The primed blood was added to the compound dish and incubated for an additional 4 h at 37 ° C. IL-1-β in supernatants was measured using an AlphLISA kit (Cat # AL220) according to the manufacturer's instructions. The compounds showed a dose-dependent increase in the production of IL-1e. EC _{50 was} determined using primed but untreated blood as a baseline.

Measurement of IL-1β production - hTRF protocol for mice.

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

Serial dilutions of compounds in DMSO were added to cells in 384-well low volume plates at 60 nl / well using an ECHO 550 acoustic dispenser (Labcyte) to achieve a final initial concentration of 50 μM in the assay, and incubated with compounds for an additional 2 h at 37 ° C at 5% CO ₂ .

After a 2-hour incubation, the cell plates were centrifuged for 5 min at 1200 rpm. Using Felix (CyBio), 8 μl of supernatant was transferred into small volume 384-well white proxy plates. (Perkin Elmer, 6008230). The hTRF Human IL-1-β Detection Kit (CISBIO, 62MIL1BPEH) was used for the analysis of the supernatant. Followed the instructions of the kit to obtain the standard curve ILlBeta (antibodies from the kit were diluted 1:40, not 1:20, as indicated in the instructions). After pooling, antibodies were added to all plates at 5 μl / well. The plates were covered and incubated at 4 ° C overnight. Plates were read on a Perkin Elmer EnVision at 665/615 nm using an hTRF laser. The data were then converted to pg / ml Il1Beta. The compounds showed a dose-dependent increase in IL-1 production.

In vitro human TLR7 and TLR8 reporter binding assays.

Logarithmically growing human HEK-Blue cells co-expressing the TLR7 or TLR8 gene and NF-kB / AP1-inducible SEAP (secreted embryonic alkaline phosphatase; Invivogen,

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San Diego, CA), the reporter gene was added to individual wells of a 384-well plate (15,000 cells / 20 μl / well) and incubated for 24 h at 37 ° C, 5% CO ₂ . Test compounds or DMSO were dispensed into individual wells the next day using an acoustic liquid dosing system (100 nl per well) and the cells were then incubated for 18 hours at 37 ° C, 5% CO ₂ . SEAP production in cells is measured using an Envision plate reader thirty minutes after the addition of freshly prepared Quanti-Blue reagent (prepared according to manufacturer's instructions; Invivogen, San Diego, CA) to reactions containing HEK-Blue TLRNf-kB- cells SEAP. All EC50 values (half maximum effective concentration) were determined using proprietary data analysis software. Normalized EC50 value = absolute value determined by setting 100% Ymax using the RLU (relative light unit) values of the reference standard for cells treated with 50 μM reference standard.

Pharmacology in vivo.

Compounds were evaluated for in vivo efficacy in preclinical syngeneic tumor models such as MC38, CT26 and 4T1, respectively. Tumor lines were implanted subcutaneously in syngeneic, immunocompetent mice. For the intratumoral (IT) route of administration of the compounds, abscopal tumor models are used. In the abscopal model, mice are injected subcutaneously with 0.1 ml of cells (1x10 ⁷ cells / ml) into the right and left flanks, respectively, using a 1 ml tuberculin syringe with a 25g needle. Animals with tumors are sorted and randomized when tumors on each side reach approximately 100 mm ³ . Compounds are administered by IT injection in the right flank at appropriate doses and frequency, either alone or in combination with checkpoint blockers such as anti-PD-1 and / or anti-CTLA4. For combination studies, checkpoint blockers are administered intraperitoneally (IP) at optimal doses and dosing frequency. Efficacy is determined by monitoring the tumor volumes of the injected and abscopal tumor, respectively.

Additional studies are carried out on abscopal tumor models to determine the PK / PD relationship, as well as to assess the profile of tumor-infiltrating lymphocytes (TIL). Other routes of administration, such as intravenous or intramuscular, are being investigated along with various dosing regimens to determine dosing routes and regimens that would provide optimal efficacy.

The above table. 1 includes biological data on compounds that have been analyzed using one or more of the aforementioned techniques.

A number of embodiments of the invention have been described. However, it will be understood that various modifications can be made without departing from the spirit and scope of the invention. Accordingly, other embodiments are within the scope of the following claims.

Claims (24)

CLAIM 1. Compound of formula (II) ^R4 (U, or its pharmaceutically acceptable salt, where R ^{1 is} independently unsubstituted C _1-6 alkyl or C (= O) R ^a ; R ^{2 is} independently H or unsubstituted C _1-6 alkyl; R ³ represents: (i) H; (ii) unsubstituted C _1-2 alkyl; (iii) XR ⁸ wherein X is unbranched ^{C1-6 alkylene and R 8} is —OH, C1-4 alkoxy, —C _1-4 haloalkoxy, CO ₂ R ^a, or —CONRcRd; (iv) (C _1-3 alkylene) (C ₆ -C ₁₀ aryl), wherein the aryl is optionally substituted with 1 to 3 substituents independently selected from C _1-6 alkyl, C _1-4 haloalkyl, C _1-4 alkoxy and C _1-4 haloalkoxy; or (v) (C _1-3 alkylene) heteroaryl having 5 to 6 ring atoms, where 1 to 4 ring atoms are each independently selected from N, N (R ^e ), O and S, and where heteroaryl is optionally substituted from 1 up to 3 substituents independently selected from C _1-6 alkyl, C _1-4 haloalkyl, C _1-4 alkoxy and C _1-4 halogen - 111 037780 alkoxy; R ⁴ and R ⁵ are each independently selected from: (i) H; (ii) halogen; (iii) - (C _0-3 alkylene) C _3-10 cycloalkyl, wherein cycloalkyl is optionally substituted with 1 to 4 independently selected Rf; (iv) - (C _0-3 alkylene) heterocyclyl having from 3 to 10 ring atoms, with from 1 to 3 ring atoms, each independently selected from N (R ^e ), O and S, wherein the heterocyclyl is optionally substituted from 1 up to 4 R ^f ; (v) - (C _0-3 alkylene) (C ₆ -C ₁₀ aryl) optionally substituted with 1 to 4 R ^g ; (vi) - (C _0-3 alkylene) heteroaryl having 5 to 10 ring atoms, with 1 to 4 ring atoms each independently selected from N, NH, O and S, wherein the heteroaryl is optionally substituted with 1 to 3 R ^g ; (vii) Cl _-6 alkyl optionally substituted with 1 to 2 independently selected R ^h ; and (viii) - (C _0-3 alkylene) C _4-10 cycloalkenyl, wherein cycloalkenyl is optionally substituted with 1 to 2 Rf; R ^a represents: (i) C _1-6 alkyl optionally substituted with 1 to 2 R ^h ; or (ii) - (C _0-3 alkylene) C _3-10 cycloalkyl, wherein cycloalkyl is optionally substituted with 1 to 2 Rf; (iii) - (C _1-3 alkylene) heterocyclyl having 3 to 10 ring atoms, wherein 1 to 3 ring atoms are each independently selected from N (R ^e ), O and S, wherein the heterocyclyl is optionally substituted from 1 to 4 independently selected R ^f ; (iv) - (C _0-3 alkylene) phenyl optionally substituted with 1 to 4 independently selected R ^g ; or (v) - (C _0-3 alkylene) heteroaryl having 5 to 10 ring atoms, with 1 to 4 ring atoms each independently selected from N, N (R ^e ), O and S, where heteroaryl is optionally substituted with 1 to 3 independently selected R ^g ; in each case, R ^c and R ^{d are} independently H or C _1-4 alkyl; in each case, R ^{e is} independently H or C _1-4 alkyl; in each occurrence, Rf is independently Cl _-6 alkyl, Cl _-4 haloalkyl, —OH, F, Cl, Cl _-4 alkoxy, C _1-4 haloalkoxy, cyano, or phenyl optionally substituted with 1 to 4 R ^g ; in each case, R ^{g is} independently halogen, cyano, Cl _-6 alkyl, Cl _-4 haloalkyl, Cl _-4 alkoxy, or C _1-4 haloalkoxy; and in each occurrence, R ^{h is} independently —OH, F, Cl _—4 alkoxy, Cl _—4 haloalkoxy, or cyano. 2. A compound according to claim 1, or a pharmaceutically acceptable salt thereof, wherein R ³ is H, unsubstituted Cl-2 alkyl or XR ⁸ , where X is unbranched Cl-2 alkylene and R ⁸ is CO2R ^a or —CONR ^c R ^d ; R ⁴ independently represents H or halogen; R ^{5 is} independently selected from: (i) H; (ii) halogen; (iii) - (C _0-3 alkylene) C _3-10 cycloalkyl, wherein cycloalkyl is optionally substituted with 1 to 4 independently selected R ^f ; (iv) - (C _0-3 alkylene) heterocyclyl having from 3 to 10 ring atoms, with from 1 to 3 ring atoms, each independently selected from N (R ^e ), O and S, wherein the heterocyclyl is optionally substituted from 1 up to 4 independently selected R ^f ; (v) - (C _0-3 alkylene) (C ₆ -C _{1 0} aryl) optionally substituted with 1 to 4 R ^g ; (v) - (C _0-3 alkylene) heteroaryl having 5 to 10 ring atoms, with 1 to 4 ring atoms each independently selected from N, NH, O and S, wherein the heteroaryl is optionally substituted with 1 to 3 R ^g ; (vi) Cl _-6 alkyl optionally substituted with 1 to 2 independently selected R ^h ; and (vii) - (C _0-3 alkylene) C _4-10 cycloalkenyl, wherein cycloalkenyl is optionally substituted with 1 to 2 Rf. 3. A compound according to claim 1, or a pharmaceutically acceptable salt thereof, wherein R ^{2 is} independently H or unsubstituted C _1-3 alkyl; R ³ is H, unsubstituted Cj-2 alkyl or XR ⁸ , wherein X is unbranched C _2-4 alkylene and R ⁸ is CO2R ^a or —CONR ^c R ^d ; R ^{5 is} independently selected from: (i) C3-6cycloalkyl optionally substituted with 1 to 2 independently selected R ^f ; (ii) phenyl optionally substituted with 1 to 3 R ^g ; (iii) heteroaryl having 5 to 6 ring atoms, with 1 to 3 ring atoms each independently selected from N, NH, O and S, where heteroaryl is optionally substituted with 1 to 3 R ^g ; (iv) Cl _-6 alkyl optionally substituted with 1 to 2 independently selected R ^h ; and (v) C _5-6 cycloalkenyl optionally substituted with 1 to 2 R ^f ; and - 112 037780 R ^a is C1-4 alkyl optionally substituted with OH, C _3-6 cycloalkyl, phenyl or heteroaryl having 5 to 6 ring atoms, where 1 to 4 ring atoms are each independently selected from N, N (R ^e ) , O and S. 4. A compound according to claim 1 or 3, or a pharmaceutically acceptable salt thereof, wherein R ^{2 is} independently H, CH ₃ or CH ₂ CH ₃ ; R ³ represents H, CH ₃ or - (CH ₂ ) ₃ C (= O) OCH ₃ ; R ^{5 is} independently CH ₃ , cyclopentyl, cyclopentenyl, phenyl, pyrazol-1-yl, or pyrazol-3-yl; and R ^a represents CH ₃ , CH ₂ CH ₃ , CH (CH ₃ ) ₂ , C (CH ₃ ) ₃ , cyclopropyl or thiazolyl. 5. Connection according to any one of paragraphs. 1-4 or a pharmaceutically acceptable salt thereof, where R ^{2 is} independently H, CH ₃ or CH ₂ CH ₃ ; R ³ represents H or CH ₃ ; R ^{5 is} independently CH ₃ , cyclopentyl, cyclopentenyl, phenyl, pyrazol-1-yl, or pyrazol-3-yl; and R ^a is CH ₃ , CH ₂ CH ₃ , CH (CH ₃ ) ₂ , C (CH ₃ ) _3, or cyclopropyl. 6. Connection according to any one of paragraphs. 1-5 or a pharmaceutically acceptable salt thereof, where R ^{1 is} independently CH ₃ , CH ₂ CH ₃ , CH (CH ₃ ) ₂ , C (CH ₃ ) _3, or C (= O) R ^a ; R ^{2 is} independently H, CH ₃ or CH ₂ CH ₃ ; R ³ represents H; R ^{5 is} independently cyclopentyl, cyclopentenyl, phenyl, pyrazol-1-yl, or pyrazol3-yl; and R ^a is CH ₃ , CH ₂ CH ₃ , CH (CH ₃ ) ₂ , C (CH ₃ ) _3, or cyclopropyl. 7. A compound according to claim 1, wherein the compound is and a pharmaceutically acceptable salt thereof. 8. A compound according to claim 1, wherein the compound is 9. A compound according to claim 1, wherein the compound is pH ₃ and a pharmaceutically acceptable salt thereof. 10. A compound according to claim 1, wherein the compound is 11. A pharmaceutical composition containing a compound or a pharmaceutically acceptable 113 037780 salt, according to any one of claims 1 to 10 and one or more pharmaceutically acceptable excipients. 12. Use of a compound or a pharmaceutically acceptable salt thereof according to any one of claims 1 to 10 as a medicament. 13. Use of the pharmaceutical composition according to claim 11 as a medicine. 14. Use of a compound, or a pharmaceutically acceptable salt thereof, according to any one of claims 1-10 for the treatment of cancer in a subject. 15. Use of a pharmaceutical composition according to claim 11 for treating cancer in a subject. 16. Use according to claim 14 or 15, wherein the cancer is selected from acute myeloid leukemia, adrenocortical carcinoma, Kaposi's sarcoma, lymphoma, anal cancer, appendix cancer, teratoid / rhabdoid tumor, basal cell carcinoma, bile duct cancer, bladder cancer , bone cancer, brain cancer, breast cancer, bronchial tumor, carcinoid tumor, heart tumor, cervical cancer, chordoma, chronic lymphocytic leukemia, chronic myeloproliferative tumor, colon cancer, colorectal cancer, craniopharyngioma, bile duct cancer, endometrial cancer , ependymoma, esophageal cancer, esthesioneuroblastoma, Ewing's sarcoma, eye cancer, fallopian tube cancer, gallbladder cancer, gastrointestinal carcinoid tumor, gastrointestinal stromal tumor, germ cell tumor, hair cell leukemia, head and neck cancer, heart cancer, liver cancer, hypopharyngeal cancer, pancreatic cancer, kidney cancer, laryngeal cancer, chronic myelogenous leukemia, lip and oral cavity cancer, lung cancer, melanoma, Merkel carcinoma, mesothelioma, oral cancer, oral cancer, osteosarcoma, ovarian cancer, penile cancer, pharyngeal cancer, prostate cancer, rectal cancer, salivary gland cancer , skin cancer, small bowel cancer, soft tissue sarcoma, testicular cancer, throat cancer, thyroid cancer, urethral cancer, uterine cancer, vaginal cancer, and vulvar cancer. 17. The use according to any one of paragraphs. 14-16, and the cancer is refractory cancer. 18. Use according to claim 16, wherein the cancer is selected from breast cancer, colon cancer, rectal cancer, colorectal cancer, pancreatic cancer and prostate cancer. 19. Use according to any one of claims 14-18, wherein the compound or composition can be administered in combination with one or more additional anti-cancer therapies. 20. Use according to claim 19, wherein the one or more additional anti-cancer therapies include surgery, radiation therapy, chemotherapy, toxotherapy, immunotherapy, cryotherapy, or gene therapy, or a combination thereof. 21. Use according to claim 19, wherein the adjunctive anticancer therapy comprises one or more agents selected from cisplatin, carboplatin, mechlorethamine, cyclophosphamide, chlorambucil, ifosfamide, oxaliplatin, azathioprine, mercaptopurine, vincristine, vinblastine, vinorelbine, tacaxel, vindesine , docetaxel, irinotecan, amsacrine, etoposide, etoposide phosphate, teniposide, actinomycin, anthracyclines, doxorubicin, daunorubicin, valrubicin, idarubicin, epirubicin, bleomycin, plikamycin, mitomycin, gupreliquin adalimumab, alemtuzumab, atlizumaba, basiliximab, belimumab, bevacizumab brentuximab vedotin, kanakinumaba, cetuximab, certolizumab pegola, daclizumab, denosumab, ekulizumaba, efalizumab, gemtuzumab, golimumab, ibritumomab tiuxetan, infliximab, ipilimumab, muromonab-CD3, natalizumab, ofatumumab, omalizumab , palivizumaba, panitumuaba, ranibizumab, rituximab, tocilizumab, tositumomab, trastuzumab, interleukin-2, indoleamine-2,3-dioxygenase, IL-10, transforming growth factor-β, CD39, CD73, adenosine-CD39-CD73 and CXCR4-CXCL12. 22. Use according to claim 19, wherein the adjunctive cancer therapy comprises one or more agents selected from urelumab, PF 05082566, MEDI6469, TRX518, varlilumab, CP 870893, pembrolizumab, nivolumab, atezolizumab, MEDI4736, avelumab, PDR161, BMS MGA271, lirilumab, IPH2201, emactuzumab, INCB024360, galunizertiba, ulocuplumab, BKT140, bavituximab, CC 90002, bevacizumab, and MNRP1685A. 23. Use according to claim 19, wherein the adjunctive cancer therapy comprises one or more agents selected from nivolumab, ipilimumab, pembrolizumab, atezolizumab, and avelumab. 24. Use according to any one of claims 12 to 22, wherein the compound can be administered intratumorally or systemically.