Immunomodulatory compound

Abstract

A compound of formula 1, [Formula 1] JPEG2025504259000021.jpg3193 During the ceremony, A 1 , CH2, CHR 4 , C.R. 4 R 5 , N.H., N.R. 4 , O or S; A 2 , A 3 , A 4 , and A 5 , CH, CR 6 or N; R 1 , R 2 , R 3 , R 4 , R 5 , R 6 is, when present, independently hydrogen, hydroxy, acetyl, halo, carboxyl; substituted or unsubstituted amino, alkyl, alkoxy, carboxyalkyl, alkylamido, acyl, acylaminoaryl, arylalkyl, heteroalkyl, heteroalkoxy, heterocarboxyalkyl, heteroacyl, heteroacylamino, heteroaryl, or heteroarylalkyl; or a substituted or unsubstituted 5-membered heterocycle, where any member of a given ring may be C, N, O, or S. Also provided is a method of treating a mammal suffering from an inflammatory disease comprising administering the compound to the mammal in a manner sufficient to reduce the severity of the inflammatory disease.

Description

[Technical field]

[0001] This application relates generally to compounds that modulate the immune system of a mammal. More specifically, compounds are provided that modulate multiple immune factors and ameliorate several diseases. [Background technology]

[0002] Dysregulation of the immune system can lead to a number of health conditions and disorders, including neurodegenerative diseases, complications resulting from viral infections, autoimmune diseases, and alterations in whole body metabolism. Current strategies for treating these conditions include blocking individual cytokines or chemokines and inhibiting key pathways related to immune activation. In many cases, these strategies result in loss of response over time or are poorly tolerated with many side effects related to widespread immune suppression. New strategies have the potential to reach millions of patients.

[0003] Recent studies have revealed the ability to regulate immune responses based on cellular metabolism. The relative balance of glucose and lipid metabolism and oxidative and anaerobic pathways differs between cells that promote inflammation and cells that limit inflammation. Therefore, there is a need for compounds that target receptors and signaling proteins in immune cells that affect this balance. The present invention provides several such compounds. Summary of the Invention

[0004] A compound of formula 1, [ka] During the ceremony, A 1 , CH2, CHR 4 , C.R. 4 R 5 , N.H., N.R. 4 , O or S; A 2 , A 3 , A 4 , and A5 , CH, CR 6 or N; R 1 , R 2 , R 3 , R 4 , R 5 , R 6 is, when present, independently hydrogen, hydroxy, acetyl, halo, carboxyl; substituted or unsubstituted amino, alkyl, alkoxy, carboxyalkyl, alkylamido, acyl, acylaminoaryl, arylalkyl, heteroalkyl, heteroalkoxy, heterocarboxyalkyl, heteroacyl, heteroacylamino, heteroaryl, or heteroarylalkyl; or a substituted or unsubstituted 5-membered heterocycle, where any member of a given ring may be C, N, O, or S.

[0005] Also provided is a method of treating a mammal suffering from an inflammatory disease comprising administering the compound to the mammal in a manner sufficient to reduce the severity of the inflammatory disease. [Brief description of the drawings]

[0006] [Figure 1A] FIG. 1 shows the structures and binding affinities of compounds NIM-1001-1 to NIM-1001-5. [Figure 1B] FIG. 1 shows the structures and binding affinities of compounds NIM-1001-6 to NIM-1001-10. [Figure 1C] FIG. 1 shows the structures and binding affinities of compounds NIM-1001-11 to NIM-1001-25. [Figure 1D] FIG. 1 shows the structures and binding affinities of compounds NIM-1001-16 to NIM-1001-20. [Figure 1E] FIG. 1 shows the structures and binding affinities of compounds NIM-1001-21 to NIM-1001-25. [Figure 1F] FIG. 1 shows the structures and binding affinities of compounds NIM-1001-26 to NIM-1001-30. [Figure 1G] FIG. 1 shows the structures and binding affinities of compounds NIM-1001-31 to NIM-1001-35. [Figure 1H] FIG. 1 shows the structures and binding affinities of compounds NIM-1001-36 to NIM-1001-39. [Figure 2A] 1 is a graph showing the reduction in spleen size following treatment of a mouse model of systemic inflammation with various compounds of Formula 1. [Figure 2B] 1 is a graph showing the reduction of CD4+IL21+ cells following treatment of a mouse model of systemic inflammation with various compounds of Formula 1. [Figure 2C] 1 is a graph showing the increase in CD25+FOXP3+ cells following treatment of a mouse model of systemic inflammation with various compounds of Formula 1. [Figure 3A] 1 is a graph showing the reduction in CD4+ cells following treatment of a mouse model of acute inflammatory bowel disease with various compounds of formula 1. [Figure 3B] 1 is a graph showing the increase in CD25+ Treg cells following treatment of a mouse model of acute inflammatory bowel disease with various compounds of formula 1. [Figure 3C] 1 is a graph showing the reduction of F4 / 80hi cells following treatment of a mouse model of acute inflammatory bowel disease with various compounds of formula 1. [Figure 4] 1 is a graph showing increased % survival following treatment of mice infected with Influenza A H1N1 with various compounds of Formula 1. [Figure 5A] 1 is a graph showing reduction in disease severity following treatment of mouse models of experimental autoimmune encephalomyelitis and multiple sclerosis with NIM-1001-8. [Figure 5B] 1 is a graph showing reduction of CD4+IL17+ cells following treatment of mouse models of experimental autoimmune encephalomyelitis and multiple sclerosis with NIM-1001-8. [Figure 5C] 1 is a graph showing reduction of GFAP+ staining cells following treatment of mouse models of experimental autoimmune encephalomyelitis and multiple sclerosis with NIM-1001-8. [Figure 6A]1 is a graph showing reduction in urinary protein levels following treatment of a mouse model of systemic lupus erythematosus (SLE) with NIM-1001-8. [Figure 6B] FIG. 1 is a graph showing an increase in splenic CD25+FOXP3+ regulatory T cells following treatment of a mouse model of systemic lupus erythematosus (SLE) with NIM-1001-8. [Figure 6C] 1 is a graph showing the reduction of splenic CD4+IL17+ cells following treatment of a mouse model of systemic lupus erythematosus (SLE) with NIM-1001-8. [Figure 7A] FIG. 1 is a graph showing the increase in time spent in light following treatment of a mouse model of Alzheimer's disease with NIM-1001-8. [Figure 7B] FIG. 1 is a graph showing neutrophil depletion following treatment of a mouse model of Alzheimer's disease with NIM-1001-8. [Figure 7C] FIG. 1 is a graph showing the reduction of CD4+IL17+ cells following treatment of a mouse model of Alzheimer's disease with NIM-1001-8. [Figure 8A] FIG. 13 is a graph showing reduction in step errors during the challenging beam test following treatment of the 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine induced model of Parkinson's disease with NIM-1001-8. [Figure 8B] FIG. 1 is a graph showing the reduction in splenic CD4+IL21+ T cells following treatment of a 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine-induced model of Parkinson's disease with NIM-1001-8. [Figure 8C] FIG. 1 is a graph showing the reduction in splenic CD4+IL17+ T cells following treatment of a 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine-induced model of Parkinson's disease with NIM-1001-8. [Figure 9A] 1 is a graph showing the reduction in splenic CD4+TNF+ T cells following treatment of an imiquimod-induced model of psoriasis with NIM-1001-8. [Figure 9B]FIG. 1 is a graph showing the reduction of splenic CD4+IL17+ cells following treatment of an imiquimod-induced model of psoriasis with NIM-1001-8. [Figure 9C] 1 is a graph showing reduction of inflammatory macrophages following treatment of an imiquimod-induced model of psoriasis with NIM-1001-8. [Figure 10] FIG. 1 is a graph showing reduction in disease severity by blinded histopathology scores of submandibular glands following treatment of a spontaneous NOD model of Sjogren's syndrome. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0007] The present invention provides compounds that have been developed by a novel medicinal chemistry approach (Example 1), screened using in silico and in vitro approaches to confirm binding, and verified to be biologically active in in vivo models of disease. The therapeutic use of these compounds may result in the induction or maintenance of beneficial responses in a variety of disease conditions, including, but not limited to, inflammatory or immune-mediated diseases such as psoriasis, systemic lupus erythematosus (SLE), multiple sclerosis, inflammatory bowel disease, diabetes or non-alcoholic steatohepatitis, infectious diseases of bacterial, fungal and viral origin, and cardiovascular and central nervous system disorders including atherosclerosis, Alzheimer's disease and Parkinson's disease.

[0008] In some embodiments, the present invention provides a compound of formula 1, [ka] During the ceremony, A 1 , CH2, CHR 4 , C.R. 4 R 5 , N.H., N.R. 4 , O or S; A 2 , A 3 , A 4 , and A 5 , CH, CR 6 or N; R 1 , R 2 , R 3 , R 4 , R 5 , R 6 is, if present, independently hydrogen, hydroxy, acetyl, halo, carboxyl; substituted or unsubstituted amino, alkyl, alkoxy, carboxyalkyl, alkylamido, acyl, acylaminoaryl, arylalkyl, heteroalkyl, heteroalkoxy, heterocarboxyalkyl, heteroacyl, heteroacylamino, heteroaryl, or heteroarylalkyl; or a substituted or unsubstituted 5-membered heterocycle, where any member of a given ring may be C, N, O, or S.

[0009] In some of these embodiments, A 1 is O, NH or CH2.

[0010] In another embodiment, A 2 is N.

[0011] In a further embodiment, A 3 is CH or N.

[0012] In a further embodiment, A 4 is N or CHCH3.

[0013] In another embodiment, A 5 is CH, CCH3, CCF3 or N.

[0014] In a further embodiment, R 1 is absent or is Cl, F or CH3.

[0015] In a further embodiment, R 2 are COOH, COOCH3, CONHCH3, CON(CH3)2, [ka] It is.

[0016] As used herein, [ka] indicates a bond covalently linking the R group to Formula 1 by crossing the bond.

[0017] In other embodiments, R 3 is COOH, COOCH2CH3, CON(CH3)2, CONHCH2CH3, [ka] It is.

[0018] In specific embodiments, the compounds are compounds NIM-1001-1 to NIM-1001-39: [ka] [ka] [ka] [ka] [ka] Either one of the following.

[0019] Any of these compounds can be prepared by one of ordinary skill in the art without undue experimentation, for example, by using the methods described in Example 2 or similar methods.

[0020] The compounds of formula 1 can be formulated alone or in combination with any other compound, e.g., another pharma- ceutically active compound, e.g., another compound of formula 1, in any compatible excipient.

[0021] Some embodiments of the present invention relate to the use of the above-mentioned compounds comprising formula 1 formulated in compositions, including pharmaceutical compositions, comprising at least one of the compounds of the present invention in a pharma-ceutically acceptable excipient. In some of these embodiments, the compounds are suitable for administration to patients by any parenteral, enteral, mucosal or transdermal route, such as oral, nasal, topical, pulmonary, transdermal or parenteral, such as rectal, subcutaneous, intravenous, intraurethral, ​​intramuscular, intranasal, ophthalmic solution or ointment, that effectively transports the compound of interest to the appropriate or desired site of action. The preparation of these formulations is within the skill of those skilled in the art. Methods for preparing formulations are described, for example, in Remington's Pharmaceutical Sciences (AR Gennaro, Ed.), 20th edition, Williams & Wilkins PA, USA (2000).

[0022] In some of these embodiments, the compound is combined with at least another active compound, for example, another compound that is effective for preventing or treating a disease, disorder or condition in a mammal.Alternatively or additionally, the composition can be formulated with at least one inactive ingredient, such as a cosolvent, a surfactant, an oil, a moisturizer, an emollient, a preservative, a stabilizer and an antioxidant, as a carrier or excipient.Any pharmacologically acceptable buffer, such as TRIS buffer or phosphate buffer, can be used.

[0023] A typical composition includes the compound of the present invention or its derivatives in association with a pharma- ceutically acceptable excipient, which may be, for example, a carrier or diluent. Such a composition may be in the form of a capsule, sachet, paper or other container. In producing the composition, conventional techniques for preparing pharmaceutical compositions may be used. For example, the compound of interest may be mixed with a carrier, diluted by a carrier, or enclosed within a carrier, which may be in the form of an ampoule, capsule, sachet, paper or other container. When the carrier serves as a diluent, it may be a solid, semi-solid, or liquid material that acts as a vehicle, excipient, or medium for the active compound. The compound of interest may be adsorbed onto a granular solid container, for example, in a sachet. Some examples of suitable carriers are water, salt solution, alcohol, polyethylene glycol, polyhydroxyethoxylated castor oil, peanut oil, olive oil, lactose, terra alba, sucrose, cyclodextrin, amylose, magnesium stearate, talc, gelatin, agar, pectin, acacia, lower alkyl ethers of stearic acid or cellulose, silicic acid, fatty acids, fatty acid amines, fatty acid monoglycerides and diglycerides, pentaerythritol fatty acid esters, polyoxyethylene, hydroxymethylcellulose and polyvinylpyrrolidone.Similarly, carriers or diluents may include any sustained release material known in the art, such as glyceryl monostearate or glyceryl distearate, alone or mixed with wax.The formulation may also include wetting agents, emulsifying and suspending agents, preserving agents, sweetening agents or flavoring agents.The formulations of the present invention may be formulated to provide quick, sustained, or delayed release of active ingredients after administration to a patient by using procedures well known in the art.

[0024] The pharmaceutical compositions may be sterilized and, if necessary, mixed with auxiliary substances, emulsifiers, salts for influencing osmotic pressure, buffers and / or coloring substances, etc., which do not deleteriously react with the active compounds.

[0025] For nasal administration, the formulation may contain the compound of interest dissolved or suspended in a liquid carrier, particularly an aqueous carrier, for aerosol application. The carrier may contain additives such as solubilizers, such as propylene glycol, surfactants, absorption enhancers, such as lecithin (phosphatidylcholine) or cyclodextrin, or preservatives, such as parabens.

[0026] To prepare topical formulation, compound of interest is put into dermatological vehicle as known in the art.The amount of compound of interest administered and the concentration of compound in topical formulation depend on the vehicle, delivery system or device selected, clinical condition of patient, side effects and the stability of compound in formulation.Therefore, physician uses appropriate preparation containing compound of interest at appropriate concentration and selects the amount of formulation administered according to clinical experience of the patient in question or similar patients.

[0027] For ophthalmic applications, the compounds of interest are formulated into solutions, suspensions, and ointments suitable for use in the eye.

[0028] For oral administration, either solid or fluid unit dosage forms can be prepared.To prepare solid compositions such as tablets, the compound of interest is mixed into the formulation with conventional ingredients such as talc, magnesium stearate, dicalcium phosphate, magnesium aluminum silicate, calcium sulfate, starch, lactose, acacia, methylcellulose, and other materials that are functionally similar to pharmaceutical diluents or carriers.

[0029] Capsules are prepared by mixing the compound of interest with an inert pharmaceutical diluent and filling the mixture into hard gelatin capsules of suitable size. Soft gelatin capsules are prepared by mechanically encapsulating a slurry of the compound of interest in an acceptable vegetable oil, light liquid petrolatum, or other inert oil. Fluid unit dosage forms for oral administration such as syrups, elixirs, and suspensions can be prepared. Water-soluble forms can be dissolved in an aqueous vehicle with sugar, aromatic flavorings, and preservatives to form syrups. Elixirs are prepared by using a hydroalcoholic (e.g., ethanol) vehicle containing suitable sweeteners such as sugar and saccharin, along with aromatic flavorings. Suspensions can be prepared using an aqueous vehicle with suspending agents such as acacia, tragacanth, methylcellulose, and the like.

[0030] Suitable formulations for parenteral use will be apparent to those of skill in the art, such as the use of appropriate injectable solutions or suspensions. Sterile formulations are suitable for a variety of topical or parenteral routes, including intradermal, intramuscular, intravascular, and subcutaneous.

[0031] In addition to the compound of interest, the compositions may contain pharma- ceutically acceptable, non-toxic carriers or diluents, including vehicles commonly used to form pharmaceutical compositions for animal or human administration, depending on the formulation and mode of delivery desired. The diluent is selected so as not to unduly affect the biological activity of the combination.

[0032] Examples of such diluents that are particularly useful for injectable formulations are water, various saline solutions, organic or inorganic salt solutions, Ringer's solution, dextrose solution, and Hank's solution. In addition, the pharmaceutical composition or formulation may also include additives such as other carriers; adjuvants; or nontoxic, nontherapeutic, nonimmunogenic stabilizers, and the like.

[0033] In addition, additives can be included in the formulation. Examples include cosolvents, surfactants, oils, moisturizers, emollients, preservatives, stabilizers and antioxidants. Any pharmacologically acceptable buffer can be used, such as tris buffer or phosphate buffer. The effective amount of diluent, additive and excipient is the amount effective to obtain a pharma-ceutically acceptable formulation in terms of solubility, biological activity, etc.

[0034] The compound of interest can be incorporated into microspheres.The compound of interest can be loaded into albumin microspheres, from which such microspheres can be collected in dry powder for nasal administration.Other materials suitable for preparing microspheres include agar, alginate, chitosan, starch, hydroxyethyl starch, albumin, agarose, dextran, hyaluronic acid, gelatin, collagen and casein.Microspheres can be produced by various processes known to those skilled in the art, such as spray drying process or emulsification process.

[0035] For example, albumin microspheres can be prepared by adding rabbit serum albumin in phosphate buffer to olive oil with stirring to produce a water-in-oil emulsion. A glutaraldehyde solution is then added to the emulsion, and the emulsion is stirred to crosslink the albumin. The microspheres can then be isolated by centrifugation, the oil removed, and the spheres washed, for example, with petroleum ether, followed by ethanol. Finally, the microspheres can be sieved, collected, and dried by filtration.

[0036] Starch microspheres can be prepared by adding a warm aqueous starch solution, such as potato starch, to a heated solution of polyethylene glycol in water with stirring to form an emulsion. Once a two-phase system is formed (with the starch solution as the internal phase), the mixture is then cooled to room temperature with continued stirring, after which the internal phase is converted into gel particles. These particles are then filtered off at room temperature and slurried in a solvent, such as ethanol, after which the particles are filtered off again and dried in air. The microspheres can be hardened by well-known cross-linking procedures, such as heat treatment, or by using chemical cross-linking agents. Suitable agents include dialdehydes, including glyoxal, malondialdehyde, succinaldehyde, adipaldehyde, glutaraldehyde and phthalaldehyde, diketones such as butadiene, epichlorohydrin, polyphosphates, and borates. Dialdehydes are used to crosslink proteins such as albumin by interaction with amino groups, diketones form Schiff bases with amino groups, and epichlorohydrin activates compounds bearing nucleophiles such as amino or hydroxyl to epoxide derivatives.

[0037] Another embodiment of the present invention is an administration scheme. The term "unit dosage form" refers to a physically discrete unit suitable as a unitary dosage for subjects, such as mammalian subjects, such as humans, dogs, cats and rodents, each unit containing a predetermined amount of active material calculated to produce the desired pharmaceutical effect in combination with the required pharmaceutical diluent, carrier or vehicle. The specifications of the unit dosage forms of the present invention are determined by and depend on (a) the unique characteristics of the active material and the particular effect to be achieved, and (b) the limitations inherent in the technology of compounding such active materials for use in humans and animals. Examples of unit dosage forms are tablets, capsules, pills, powder packets, wafers, suppositories, granules, cachets, tezapone ful, tezapone ful, drops, ampoules, vials, aerosols with metered release, separate multiples of any of the above, and other forms described herein. The composition can be included in a kit, which can contain one or more unit dosage forms of the composition and instructions for use in treating one or more of the disorders described herein.

[0038] Slow or extended release delivery systems, including either systems using numerous biopolymers (biological systems), liposomes, colloids, resins, and other polymeric delivery systems, or compartmentalized reservoirs, can be utilized with the compositions described herein to provide a continuous or long-term source of therapeutic compounds. Such slow release systems are applicable to formulations for delivery by topical, intraocular, oral, and parenteral routes.

[0039] An effective amount of the compound of interest is used for treatment. The dosage of the compound used according to the present invention varies depending on the compound and the disease, disorder or condition being treated, such as the age, weight and clinical condition of the recipient patient. Other factors include the route of administration, the patient, the patient's medical history, the severity of the disease process, and the efficacy of the particular compound. The dosage should be sufficient to improve the symptoms or signs of the disease being treated without causing unacceptable toxicity to the patient. In general, an effective amount of a compound is an amount that provides either a subjective relief of symptoms or an objectively identifiable improvement as indicated by the clinician or other qualified observer. Those skilled in the art can determine the appropriate dosage for any particular use of the route of administration without undue experimentation.

[0040] In some embodiments, the compound can prevent or reduce the severity of an inflammatory disease when administered to a mammal with the inflammatory disease. Any inflammatory disease can be treated along with the disease. Non-limiting examples of inflammatory diseases include: systemic inflammation, inflammatory bowel disease, viral infections that cause inflammation, autoimmune encephalomyelitis, multiple sclerosis, systemic lupus erythematosus (SLE), Alzheimer's disease, Parkinson's disease, psoriasis, Sjogren's disease, encephalitis, myelitis, meningitis, arachnoiditis, PNS, neuritis, lacrimitis, scleritis, episcleritis, keratitis, retina. inflammation, chorioretinitis, blepharitis, conjunctivitis, uveitis, otitis externa, otitis media, labyrinthitis, mastoiditis, carditis, endocarditis, myocarditis, pericarditis, vasculitis, arteritis, phlebitis, capillaritis, sinusitis, rhinitis, pharyngitis, laryngitis, tracheitis, bronchitis, bronchiolitis, bronchopneumonia, pleuritis, mediastinitis, stomatitis, gingivitis, gingivostomatitis, glossitis, tonsillitis, sialadenitis / parotitis, cheilitis, dentition Meningitis, jaw inflammation, gastritis, gastroenteritis, enteritis, colitis, enteritis, duodenitis, ileitis, appendicitis, proctitis, hepatitis, ascending cholangitis, cholecystitis, pancreatitis, peritonitis, dermatitis, folliculitis, cellulitis, hidradenitis, arthritis, dermatomyositis, myositis, synovitis / tendinitis, bursitis, enthesitis, fasciitis, capsulitis, epicondylitis, tendonitis, seborrheic dermatitis, osteochondritis: osteitis / osteomyelitis, spondylitis, periostitis, chondritis , nephritis, glomerulonephritis, pyelonephritis, ureteritis, cystitis, urethritis, salpingitis, salivary gland inflammation, endometritis, epididymitis, cervicitis, vaginitis, vulvitis, mastitis, orchitis, epididymitis, prostatitis, seminal vesiculitis, balanitis, posterior orchitis, balanoposthorchitis, chorioamnionitis, mycobacteritis, oophoritis, insulitis, uterine hypophysitis, thyroiditis, parathyroiditis, adrenalitis, lymphangitis, and lymphadenitis.

[0041] In some embodiments, the inflammatory disease is systemic inflammation, inflammatory bowel disease, a viral infection that causes inflammation, autoimmune encephalomyelitis, multiple sclerosis, systemic lupus erythematosus (SLE), Alzheimer's disease, Parkinson's disease, psoriasis, or Sjogren's disease.

[0042] In some embodiments where the inflammatory disease is systemic inflammation, administration of the compound to the mammal reduces spleen size, reduces the percentage of CD4+IL21+ cells in the spleen, and / or increases the percentage of CD25+FOXP3+ cells in the spleen in the mammal. See, e.g., Example 3 below.

[0043] In various embodiments where the inflammatory disease is acute inflammatory bowel disease, administration of the compound reduces the percentage of CD4+ cells in the lamina propria of the mammal's colon, increases the percentage of CD25+ Treg cells in the mammal's colon, and / or reduces F4 / 80hi macrophages in the mammal. See, e.g., Example 4 below.

[0044] In certain embodiments, where the inflammatory disease is a viral infection that causes inflammation, administration of the compound reduces mortality in a population of mammals infected with the virus, reduces the spread of non-lethal viruses, and / or reduces symptoms from the viral infection in the mammal. The virus in these embodiments is any virus now known or later discovered that can infect a mammal. Non-limiting examples of viruses in these embodiments include herpes viruses (e.g., human cytomegalovirus (HCMV), herpes simplex virus 1 (HSV-1), herpes simplex virus 2 (HSV-2), varicella zoster virus (VZV), Epstein-Barr virus), influenza A virus, or picornaviruses such as Coxsackievirus B3 (CVB3). Other viruses include, but are not limited to, hepatitis B virus, HIV, poxviruses, hepadaviruses, retroviruses, and flaviviruses, togaviruses, coronaviruses, hepatitis D virus, orthomyxoviruses, paramyxoviruses, rhabdoviruses, bunyaviruses, filo viruses, adenoviruses, human herpesvirus type 8, human papillomavirus, BK virus, JC virus, smallpox, hepatitis B virus, human bocavirus, parvovirus B19, human astrovirus, Norwalk virus, virus, coxsackievirus, hepatitis A virus, poliovirus, rhinovirus, severe acute respiratory syndrome (SARS) virus, hepatitis C virus, yellow fever virus, dengue virusExamples of viruses include RNA viruses such as the Hepatitis virus, West Nile virus, Rubella virus, Hepatitis E virus, and human immunodeficiency virus (HIV). In some cases, the virus is an enveloped virus. Examples include, but are not limited to, viruses that are members of the hepadnavirus family, herpesvirus family, iridovirus family, poxvirus family, flavivirus family, togavirus family, retrovirus family, coronavirus family, filovirus family, rhabdovirus family, bunyavirus family, orthomyxovirus family, paramyxovirus family, and arenavirus family. Other examples include, but are not limited to, the hepadnaviruses Hepatitis B virus (HBV), woodchuck hepatitis virus, ground squirrel squirrel (Hepadnaviridae) hepatitis virus, duck hepatitis B virus, heron hepatitis B virus, herpes viruses, herpes simplex virus (HSV) types 1 and 2, varicella-zoster virus, cytomegalovirus (CMV), human cytomegalovirus (HCMV), mouse cytomegalovirus (MCMV), guinea pig cytomegalovirus (GPCMV), Epstein-Barr virus (EBV), human herpesvirus 6 (HHV variants A and B), human herpesvirus 7 (HHV-7), human herpesvirus 8 (HHV-8), Kaposi's sarcoma-associated herpesvirus (KSHV), B virus, poxvirus vaccinia virus,virus, variola virus, smallpox virus, monkeypox virus, cowpox virus, camelpox virus, ectromelia virus, mousepox virus, rabbitpox viruses, raccoonpox viruses, molluscum contagiosum virus, orf virus, milker's nodes virus, bovin papullar stomatitis virus, sheeppox virus, goatpox virus, lumpy skin disease virus, fowlpox virus virus, canarypox virus, pigeonpox virus, sparrowpox virus, myxoma virus, hare fibroma virus, rabbit fibroma virus, squirrel fibroma virus, swinepox virus, tanapox virus, yabapox virus, flavivirus dengue virus, hepatitis C virus (HCV), GBV hepatitis virus (GBV-A, GBV-B, and GBV-C), West Nile virus, yellow fever virus, St. Louis encephalitis virus, Japanese encephalitis virusvirus, Powassan virus, tick-borne encephalitis virus, Kyasanur Forest disease virus, Togavirus, Venezuelan equine encephalitis (VEE) virus, chikungunya virus, Rose River virus, Mayaro virus, Sindbis virus, rubella virus, retroviruses human immunodeficiency virus (HIV) types 1 and 2, human T-cell leukemia virus (HTLV) types 1, 2 and 5, mouse mammary tumor virus (MMTV), Rous sarcoma virus (RSV), lentiviruses, filoviruses Ebola virus, Marburg virus, virus, metapneumoviruses (MPV), such as human metapneumovirus (HMPV), rhabdovirus rabies virus, vesicular stomatitis virus, Bunyavirus, Crimean-Congo hemorrhagic fever virus, Rift Valley fever virus, La Crosse virus, Hantaan virus, Orthomyxovirus, influenza viruses (types A, B, and C), paramyxovirus, parainfluenza virus (PIV types 1, 2, and 3), respiratory syncytial virus,virus (types A and B), measles virus, mumps virus, Arenavirus, lymphocytic choriomeningitis virus, Junin virus, Machupo virus, Guanarito virus, Lassa virus, Ampari virus, Flexal virus, Ippy virus, Mobala virus, Mopeia virus, Latino virus, Parana virus, Pichinde virus, Punta toro virus (PTV), Tocaribe virus, and Tamiami virus. In some embodiments, the virus is an influenza A virus, such as an H1N1 strain. See, e.g., Example 5 below.

[0045] In some embodiments where the inflammatory disease is autoimmune encephalomyelitis or multiple sclerosis, administration of the compound reduces the percentage of CD4+IL17+ T cells in the spinal cord of the mammal, reduces astrocytes or activated glia in the spinal cord of the mammal, and / or reduces GFAP positive staining in the spinal cord of the mammal. See, e.g., Example 6 below.

[0046] In various embodiments, the inflammatory disease is systemic lupus erythematosus (SLE), lupus nephritis, cutaneous lupus, or lupus with central nervous system (CNS) involvement, and the compound reduces urinary protein levels in the mammal, increases splenic CD25+FOXP3+ regulatory T cells in the mammal, and / or decreases splenic CD4+IL17+ cells in the mammal. See, e.g., Example 7 below.

[0047] In certain embodiments where the inflammatory disease is Alzheimer's disease, administration of the compound reduces an anxiety response by the mammal, reduces neutrophils in the mammal, and / or reduces CD4+IL17+ cells in the mammal. See, e.g., Example 8 below.

[0048] In some embodiments where the inflammatory disease is Parkinson's disease, administration of the compound improves gait in the mammal, reduces splenic CD4+IL21+ cells in the mammal, and / or reduces splenic CD4+IL17+ cells in the mammal. See, e.g., Example 9 below.

[0049] In various embodiments where the inflammatory disease is psoriasis, administration of the compound reduces splenic CD4+TNF+ T cells in the mammal, reduces splenic CD4+IL17+ in the mammal, and / or reduces inflammatory macrophages in the mammal. See, e.g., Example 10 below.

[0050] In certain embodiments, where the inflammatory disease is Sjogren's syndrome, administration of the compound reduces disease severity and histopathology of the lacrimal and / or salivary glands in a mammal. See, e.g., Example 11 below.

[0051] The pharmaceutical compositions described above can be formulated for administration to any mammal, including, but not limited to, livestock (e.g., cows, pigs, goats, sheep, llamas, alpacas, minks, etc.) and domestic animals (e.g., dogs, cats, mice, rats, gerbils, guinea pigs, ferrets, etc.) and humans.

[0052] Methods of treating mammals using the pharmaceutical compositions of Formula 1 above are also provided herein.

[0053] Thus, in some embodiments, a method of treating a mammal suffering from an inflammatory disease is provided, comprising administering to the mammal a compound identified above in a manner sufficient to reduce the severity of the systemic inflammation. In some embodiments, the inflammatory disease is systemic inflammation, inflammatory bowel disease, a viral infection causing inflammation, autoimmune encephalomyelitis, multiple sclerosis, systemic lupus erythematosus (SLE), Alzheimer's disease, Parkinson's disease, psoriasis, or Sjogren's disease, as described above.

[0054] These methods can be used to treat any mammal, including, but not limited to, livestock (e.g., cows, pigs, goats, sheep, llamas, alpacas, minks, etc.) and farm animals (e.g., dogs, cats, mice, rats, gerbils, guinea pigs, ferrets, etc.) and humans. Additionally, these methods can utilize any mode of administration, including any parenteral, enteral, transmucosal, or transdermal administration, as described above. EXAMPLES

[0055] Preferred embodiments are described in the following examples. Other embodiments within the scope of the claims herein will be apparent to those skilled in the art from consideration of the specification or practice of the invention disclosed herein. It is intended that the specification, together with the examples, be considered exemplary only, with the scope and spirit of the invention being indicated by the claims which follow the examples.

[0056] Example 1. Molecular docking of NIM-1001 compound method A lattice space was defined. The three-dimensional structures of the ligands with defined atomic charges were docked into the defined regions. The binding affinities were calculated based on the interactions between the ligands and the protein structure. Potential ligands were analyzed and ranked by raw binding affinity, molecular weight normalized binding affinity, and binding pose, among other criteria.

[0057] result Thirty-nine exemplary docked structures, designated NIM-1001-1 through NIM-1001-39, and the resulting binding affinities are shown in Figures 1A through 1H. The docked structures were identified to have binding affinities ranging from -7.4 kcal / mol to -9.4 kcal / mol. Structures with the most favorable binding affinities included NIM-1001-2, NIM-1001-27, and NIM-1001-33, all of which had binding affinities greater than 9.

[0058] Example 2. Synthesis of NIM-1001-8 The synthesis of methyl (E)-2-chloro-6-((5-(3-ethoxy-3-oxoprop-1-en-1-yl)-4-methylpyridin-3-yl)oxy)isonicotinate (NIM-1001-8) was a four-step process.

[0059] Hydrobromic acid was added to a solution of 3-bromo-5-methoxy-4-methylpyridine in acetic acid. The mixture was stirred at 120° C. for 3 days, and HBr was added every 24 hours. After completion of the reaction, the reaction mixture was neutralized with NaOH and extracted with ethyl acetate. The product was isolated as 5-bromo-4-methylpyridin-3-ol.

[0060] Cesium carbonate and methyl 2,6-dichloropyridine-4-carboxylate were added to a solution of 5-bromo-4-methylpyridin-3-ol in DMF and stirred at 120° C. for 4 hours. After completion of the reaction, the reaction mixture was filtered through a bed of Celite. The filtrate was evaporated under reduced pressure to give the crude product, which was carried on to the next step without purification.

[0061] Concentrated sulfuric acid was added to the solution of the crude product in methanol and stirred at 70° C. for 16 hours. After completion of the reaction, the solvent was evaporated under reduced pressure and extracted with ethyl acetate. The crude product was isolated and purified to give methyl (E)-2-chloro-6-((5-bromo-4-methylpyridin-3-yl)oxy)isonicotinate.

[0062] Triethylamine, ethyl acrylate, and tris(o-tolyl)phosphine were added to methyl (E)-2-chloro-6-((5-bromo-4-methylpyridin-3-yl)oxy)isonicotinate in acetonitrile. It was stirred under nitrogen atmosphere for 10 minutes, then palladium(II) acetate was added and stirred at 90° C. for 18 hours. After completion of the reaction, the solvent was evaporated under reduced pressure to obtain the crude product. The crude product was subjected to purification to obtain the desired product NIM-1001-8 as a solid. 1 H NMR (400 MHz, DMSO-d6): δ 8.83 (s, 1H), 8.46 (s, 1H), 7.83 (d, J = 16.00 Hz, 1H), 7.62 (s, 1H), 7.53 (s, 1H), 6.75 (d, J = 16.00 Hz, 1H), 4.23 (q, J= 7.20 Hz, 2H), 3.92 (s, 3H), 2.22 (s, 3H), 1.28 (t, J = 6.80 Hz, 3H).

[0063] Example 3. Use of NIM-1001 compound in a mouse model of systemic inflammation Introduction Systemic inflammation may result from infectious diseases, loss of self-tolerance and other unresolved perturbations to the immune system. The bm12 model is induced by transfer of lymphocytes from bm12 mice into C57BL / 6 mice. Due to mismatch in MHCII amino acid sequences, antigen-presenting cells in the recipient mice induce alloactivation of donor lymphocytes. This alloactivation leads to a marked proliferation of donor T follicular helper cells and recipient GC B and plasma cells. This adoptive transfer model has been used as a model of graft-versus-host disease and also shares immunological changes with SLE-like disease, which presents with antinuclear antibodies, IgG deposits in the kidney and type I IFN production.

[0064] method Mouse model. Wild type (WT) were used on a C57BL / 6 background. Bm12 mice were used on a C57BL / 6 background with a mutation in the MHCII allele. Spleens and major lymph nodes were collected from bm12 mice in RPMI and disrupted to obtain a cell suspension. Red blood cells were lysed. Cells were counted and resuspended in sterile PBS. 2.5×10 7 Cells / mouse were injected intraperitoneally into WT recipients on project day 0. Treatments or vehicle controls were administered daily by oral gavage starting one week after transfer in a 0.5% methylcellulose suspension to provide 20 mg / kg / day.

[0065] Immunological analysis. Spleens were excised, weighed, disrupted and filtered to obtain a cell suspension. Red blood cells were lysed. Cells were labeled with a mixture of extracellular (CD45, CD3, CD4, CD8, CD25) and intracellular (FOXP3, IL21, IL10) antibodies in sequential live staining in 96-well plates in preparation for flow cytometry. Data were captured on a BD FACS Celesta and analyzed using FacsDiva.

[0066] result Oral treatment with NIM-1001-2, NIM-1001-8, NIM-1001-9 and NIM-1001-12 significantly reduced spleen size in proportion to total body weight (Figure 2A). NIM-1001-4, NIM-1001-5 and NIM-1001-10 reduced normalized spleen size to a lesser extent compared to the vehicle-treated group. All treatments tested reduced the percentage of CD4+IL21+ cells in the spleen compared to vehicle (Figure 2B). NIM-1001-2, NIM-1001-4, NIM-1001-8, NIM-1001-9 and NIM-1001-12 significantly increased the percentage of CD25+FOXP3+ regulatory T cells in the spleen compared to vehicle (Figure 2C). NIM-1001-4 and NIM-1001-8 induced the greatest increases across the treatments tested.

[0067] Example 4. Use of NIM-1001 compound in an acute inflammatory bowel disease model Introduction Inflammatory bowel disease is a multifactorial disease involving epithelial barrier dysfunction, perturbed interactions with the microbiota, and persistent inflammation of the intestinal mucosa. The dextran sulfate sodium (DSS) model provides a model system in which the epithelial barrier is disrupted in the distal gastrointestinal tract, allowing bacterial translocation and the generation of an immune response.

[0068] method DSS model. Mice were given DSS in drinking water for 7 days. At the start of the project, mice were 8 weeks old and treatment began 24 hours after placement in DSS. Mice were weighed and scored daily for symptoms of disease (diarrhea, rectal bleeding, rectal inflammation, overall behavior). Treatments were prepared in 0.5% methylcellulose (12-15 cP) solution. The dose used was 10 mg / kg delivered once daily.

[0069] Flow cytometry. Colons were harvested in RPMI / FBS buffer containing collagenase (300 U / mL) and DNase (50 U / mL) for digestion. Tissues were digested for 60 min with agitation at 37°C. The resulting cell suspension was filtered through a 100 μm strainer, centrifuged (300×g, 8 min), and washed with fresh RPMI. After filtering the resulting single cell suspension, immune cells were purified by a Percoll gradient of cell-containing 40% Percoll overlaid on a 70% Percoll solution. After centrifugation, the interface was collected and washed to obtain an enriched colonic lamina propria cell fraction. Cells were labeled with a mixture of extracellular (CD45, CD3, CD4, CD8, CD25, F4 / 80, CD11b, MHCII) antibodies in sequential live staining in 96-well plates. Data were acquired using a FACS Celesta flow cytometer equipped with FACSDiva software.

[0070] result Oral treatment with NIM-1001-2, NIM-1001-8, NIM-1001-11 and NIM-1001-12 reduced the percentage of CD4+ T cells in the colonic lamina propria compared to the vehicle-treated group (Figure 3A). Each NIM-1001 compound tested also significantly increased the percentage of CD25+ Tregs in the colon compared to vehicle (Figure 3B). Each NIM-1001 compound reduced F4 / 80hi macrophages compared to vehicle, with a significant reduction induced by NIM-1001-8, NIM-1001-11 and NIM-1001-12 (Figure 3C).

[0071] Example 5. Use of NIM-1001 compounds in model viral infections Introduction Viral infections remain a major concern in global public health. In particular, respiratory viral infections are a common cause of global pandemics with rapid emergence resulting in a lack of first-line treatment options. After a replication phase that generally ends a few days after exposure, the main cause of tissue damage, morbidity and mortality is an unresolved immune response. As a result, host-targeted therapy can serve as a valuable tool to combat emergent viral threats.

[0072] method Wild-type C57BL / 6 mice were anesthetized by isoflurane inhalation. Mice were anesthetized with 2×10 4 Mice were infected intranasally with influenza A (H1N1) at a challenge titer of TCID50 / mL. Mice were treated with NIM-1001-2, NIM-1001-8 or NIM-1001-9 at a dose of 25 mg / kg daily by oral gavage. Mice were monitored daily.

[0073] result The results are shown in Figure 4. Vehicle treatment resulted in mortality beginning on day 6 and reaching 25% survival by day 10. Oral treatment with NIM-1001-2 and NIM-1001-9 delayed first death by 1 day and resulted in 2 / 3 survival in each group at day 10. Oral treatment with NIM-1001-8 delayed first death by 2 days and resulted in 83% survival at day 10.

[0074] Example 6. Use of NIM-1001-8 in an Experimental Autoimmune Encephalomyelitis Model Introduction Multiple sclerosis is an inflammatory disease in which the immune system reacts to the central nervous system, resulting in relapsing and remitting or progressive damage. This neurological damage results in motor control disorders, physical disability, vision loss, depression and pain, among other symptoms. A common immunological event in many patients with MS is an increase in the number or activity of Th17 cells, especially in relapsing and remitting forms of the disease. Meanwhile, hyperactivation of microglia contributes to neurological damage.

[0075] method Mouse model. C57BL6 mice were challenged at 10-16 weeks of age by MOG immunization in complete Freund's adjuvant. MOG emulsion was administered to each mouse at 100 μL / site in the neck region and posterior flank. Ten days later, spleens were harvested. Isolated splenocytes were cultured for 3 days in the presence of MOG, IL-12 and anti-interferon gamma. Splenocytes were then washed and injected into recipient mice at approximately 20 million cells / mouse. Oral treatment with NIM-1001-8 (20 mg / kg) or vehicle was initiated 7 days after transfer. Mice were scored daily for disease activity (coordination, gait, paralysis) (0-3).

[0076] Flow cytometry. Spinal cords were excised and digested with papain and DNase. Immune cells were purified by Percoll gradient. Cells were labeled with a mixture of extracellular (CD45, CD3, CD4, CD8, CD25) and intracellular (IL17) antibodies in sequential live staining in 96-well plates in preparation for flow cytometry. Data were captured on a BD FACS Celesta and analyzed using FacsDiva.

[0077] Histology. Sections of spinal cord from the lumbar region were extracted and fixed in formalin. Samples were embedded in paraffin and sectioned. Samples were stained for the presence of glial fibrillary acidic protein (GFAP) by immunohistochemistry. Slides were evaluated microscopically and scored (0-3) for the intensity and frequency of GFAP-positive cells. GFAP is a common protein marker for astrocytes in the central nervous system.

[0078] result Oral NIM-1001-8 resulted in an improvement in overall disease severity throughout the course of the experiment (Figure 5A). NIM-1001-8 reduced the percentage of CD4+IL17+ T cells in the spinal cord (Figure 5B) and reduced the presence of GFAP-positive staining in the spinal cord (Figure 5C).

[0079] Example 7. Use of NIM-1001-8 in a genetic mouse model of SLE Introduction Systemic lupus erythematosus (SLE) is a systemic autoimmune disease that can cause kidney, cardiovascular and joint damage. SLE is the result of a complex interplay of genetic factors that results in an immunological disease that is primarily manifested through the production of autoantibodies. The MRL / lpr model is a spontaneous genetic model induced by polymorphisms in Fas / FasL. Due to these mutations, MRL / lpr mice exhibit increased lymphoproliferation, splenomegaly, glomerulonephritis and enlarged lymph nodes. The MRL / lpr model remains an important preclinical model due to the prevalence of Fas / FasL polymorphisms in SLE-susceptible individuals and the presence of double-negative T cells in human SLE.

[0080] method Mouse Model. Female MRL / lpr mice were obtained at 8 weeks of age. Mice were followed for 4 weeks for the development of proteinuria. Mice with a proteinuria score > 2 were randomized into the project at 12 weeks of age. Treatment with NIM-1001-8 or vehicle control was administered daily by oral gavage in a 0.5% methylcellulose suspension starting at 12 weeks of age.

[0081] Immunological analysis. Urine was collected for assay of protein content to test renal function. Spleens were excised, disrupted and filtered to obtain a cell suspension. Red blood cells were lysed. Cells were labeled with a mixture of extracellular (CD45, CD3, CD4, CD8, CD25) and intracellular (FOXP3, IL17) antibodies in sequential live staining in 96-well plates in preparation for flow cytometry. Data was captured on a BD FACS Celesta and analyzed using FacsDiva.

[0082] result Urinary protein levels were reduced by more than 2-fold with NIM-1001-8 treatment compared to vehicle (Figure 6A). NIM-1001-8 treatment resulted in a significant increase in CD25+FOXP3+ regulatory T cells (Figure 6B) and a significant decrease in CD4+IL17+ cells (Figure 6C). Statistical significance (P<0.05) is indicated with an asterisk.

[0083] Example 8. Use of NIM-1001-8 in a genetic model of Alzheimer's disease Introduction Alzheimer's disease is one of the most common neurodegenerative diseases that leads to memory loss and overall cognitive decline. The pathogenesis of the disease involves immune, metabolic and neurological factors that lead to the formation of amyloid-β deposits or plaques and tau tangles. There are numerous mouse models that spontaneously develop similar neurological pathologies and cognitive impairments.

[0084] method Mouse Model. Genetically modified mice that spontaneously develop cognitive impairment at 4 months of age and histological signs of disease at 6 months of age were randomized into the project at 24 weeks of age. Mice were orally treated with vehicle or NIM-1001-8 (10 mg / kg) for 8 weeks. After 8 weeks of treatment, mice were assessed using the light-dark test to measure the time spent in the light compared to the dark, which is a measure of the anxiety response. Genetically modified mice were compared to age-matched wild-type mice (negative control).

[0085] Flow cytometry. Brains were harvested in RPMI / FBS buffer containing collagenase (300 U / mL) and DNase (50 U / mL) for digestion. Tissues were digested for 30 min with agitation at 37°C. The resulting cell suspension was filtered through a 100 μm strainer, centrifuged (300×g, 8 min), and washed with fresh RPMI. After filtering the resulting single cell suspension, immune cells were purified by Percoll gradient of cell-containing 40% Percoll overlaid on 70% Percoll solution. After centrifugation, the interface was collected and washed to obtain enriched immune cell fractions. Cells were labeled with a mixture of extracellular (CD45, CD3, CD4, CD8, CD11b, Gr1) and intracellular (IL17) antibodies in sequential live staining in 96-well plates. Data were acquired using a FACS Celesta flow cytometer equipped with FACSDiva software.

[0086] result Oral NIM-1001-8 treatment significantly increased the time spent in the light compared to vehicle and restored it to equal time to the negative control group (Figure 7A). Immunologically, NIM-1001-8 reduced the percentage of neutrophils (Figure 7B) and CD4+IL17+ T cells (Figure 7C) in the brain compared to vehicle. Statistical significance (P<0.05) is indicated with an asterisk.

[0087] Example 9. Use of NIM-1001-8 in a mouse model of Parkinson's disease Introduction Parkinson's disease is a common neurodegenerative condition that causes tremors and loss of motor control. The disease is caused by damage to dopaminergic neurons, particularly in the midbrain. Mouse models of the disease exist that cause damage to these same neurons by administration of various chemicals.

[0088] method Mouse model. Mice were administered intraperitoneal injections of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) and probenecid three times a week for four weeks. Oral treatment with vehicle or NIM-1001-8 (25 mg / kg) was initiated once a day during the second week of injections. After four weeks of injections, motor control was assessed by counting the number of step errors during a challenging beam test. Mice challenged with MPTP were compared to mice challenged with sham injections (negative control).

[0089] Flow cytometry. Spleens were excised, disrupted and filtered to obtain a cell suspension. Red blood cells were lysed. Cells were labeled with a mixture of extracellular (CD45, CD3, CD4, CD8, CD25) and intracellular (IL21, IL17) antibodies in sequential live staining in 96-well plates in preparation for flow cytometry. Data were captured on a BD FACS Celesta and analyzed using FacsDiva.

[0090] result Oral NIM-1001-8 significantly reduced the number of step errors during the challenging beam test compared to vehicle-treated mice, and almost completely abolished the effects of MPTP when compared to the negative control (Figure 8A). NIM-1001-8 also reduced systemic inflammation, accompanied by a significant reduction in CD4+IL21+ (Figure 8B) and CD4+IL17+ (Figure 8C) T cells in the spleen. Statistical significance (P<0.05) is indicated with an asterisk.

[0091] Example 10. Use of NIM-1001-8 in a psoriasis model Introduction Psoriasis (PsO) is a common dermatological condition in which increased immune system activation occurs in the skin. PsO can result in the formation of plaques and thickened areas of skin, itching, and generalized discomfort. Immunologically, PsO is generally thought to be driven by Th17 cells, and IL-17 / IL-23 treatment has provided some therapeutic benefit.

[0092] method IMQ-induced model. C57BL / 6 mice were anesthetized, shaved, and briefly exposed to a superficial depilatory cream on the back. Mice were given 3 days to recover from the procedure before entering the study. After 3 days, mice were challenged daily with approximately 60 mg of 0.5% imiquimod cream by applying the cream to the shaved area. Oral treatment with vehicle or NIM-1001-8 (20 mg / kg) was administered daily.

[0093] Analysis. Spleens were excised and disrupted with a microscope slide. Red blood cells were hypotonically lysed from the resulting suspension. Samples were filtered, washed, and centrifuged before staining. Cells were labeled with a mixture of extracellular (CD45, CD3, CD4, CD8, CD25, F4 / 80, CD11b, MHCII) and intracellular (TNF, IL17) antibodies in sequential live staining in 96-well plates in preparation for flow cytometry. Data were captured on a BD FACS Celesta and analyzed using FacsDiva.

[0094] result Oral NIM-1001-8 significantly reduced the percentage of TNF+ (FIG. 9A) and IL17+ (FIG. 9B) CD4+ T cells in the spleen compared to vehicle controls. Furthermore, NIM-1001-8 reduced the percentage of F4 / 80hi inflammatory macrophages (FIG. 9C). Statistical significance (P<0.05) is indicated with an asterisk.

[0095] Example 11. Use of NIM-1001-8 in a Sjogren's syndrome model Introduction Sjögren's syndrome (SjS) is an autoimmune disease that primarily affects the lacrimal and salivary glands. SjS is generally due to the formation of autoantibodies against Ro and / or La, leading to focal infiltration of exocrine glands and disorganization of acinar and ductal epithelial cells. Immunologically, SjS is associated with dysfunctional regulatory CD4+ T cells and the proliferation of T helper 1 and T follicular helper cell subsets. Nonobese diabetic (NOD) mice have been used as a model for Sjögren's syndrome, as female mice develop salivary gland disease and male mice develop lacrimal gland disease spontaneously before the onset of hyperglycemia.

[0096] method NOD Model. Ten-week-old female NOD mice were treated with vehicle or NIM-1001-8 (2 or 50 mg / kg) daily by oral gavage for 4 weeks. After the end of the treatment period, submandibular glands were excised and fixed in formalin. Mammary glands were processed and H&E stained slides were prepared. Slides were scored by a blinded observer on a composite scale of 0 to 4 encompassing levels of inflammation and disease.

[0097] result Oral NIM-1001-8 significantly reduced the histological severity of disease in the submandibular gland at both 2 and 50 mg / kg compared to vehicle controls (Figure 10). Statistical significance (P<0.05) is indicated with an asterisk.

[0098] References Remington's Pharmaceutical Sciences (AR Gennaro, Ed.), 20th edition, Williams & Wilkins PA, USA (2000) U.S. Patent Application Publication No. 2013 / 61893613 U.S. Patent No. 8,859,546 U.S. Patent No. 7,396,943 PCT Patent Application Publication WO 2005 / 012298 PCT Patent Application Publication WO 2015 / 061247 PCT Patent Application Publication WO 2021 / 129817

[0099] In view of the above, it will be seen that the several objects of the invention are achieved and other advantageous objects attained.

[0100] Because various changes can be made in the above methods and compositions without departing from the scope of the invention, it is intended that all matter contained in the above description and shown in the accompanying drawings shall be interpreted in an illustrative and not a limiting sense.

[0101] All references cited herein, including but not limited to patent publications and non-patent publications, and the references cited therein, are hereby incorporated by reference. The description of references herein is intended merely to summarize the assertions made by the authors and is not an admission that any reference constitutes prior art. Applicants reserve the right to challenge the accuracy and pertinence of the cited references.

[0102] As used herein, in certain embodiments, the term "about" or "approximately" when preceding a numerical value indicates a range of values ​​plus or minus 10%. When a range of values ​​is provided, it is understood that each intervening value, to the tenth of the unit of the lower limit, between the upper and lower limits of that range, and any other stated or intervening value within that stated range, unless the context clearly dictates otherwise, is encompassed within the disclosure. The upper and lower limits of these smaller ranges may be independently included in the smaller ranges, and are also encompassed within the disclosure, subject to any specifically excluded limit in the stated range. When a stated range includes one or both of the limits, ranges excluding one or both of those included limits are also included in the disclosure.

[0103] The indefinite articles "a" and "an" as used in the specification and embodiments, unless expressly indicated otherwise, should be understood to mean "at least one."

[0104] The phrase "and / or" as used in the present specification and embodiments should be understood to mean "either or both" of the elements so conjoined, i.e., elements that are conjunctive in some cases and disjunctive in other cases. Multiple elements listed with "and / or" should be interpreted in the same manner, i.e., "one or more" of the elements so conjoined. Other elements may optionally be present other than the elements specifically identified by the "and / or" clause, whether related to those elements specifically identified or not. Thus, as a non-limiting example, a reference to "A and / or B", when used in conjunction with open-ended language such as "comprises", can refer in one embodiment to A only (optionally including elements other than B); in another embodiment to B only (optionally including elements other than A); in yet another embodiment to both A and B (optionally including other elements); and so forth.

[0105] As used herein and in the embodiments, "or" should be understood to have the same meaning as "and / or" defined above. For example, when separating items in a list, "or" or "and / or" should be interpreted as inclusive, i.e., including at least one of, but also including two or more of, a number or list of elements, and optionally including additional unlisted items. Only terms clearly indicated to the contrary, such as "only one of" or "exactly one of" or, when used in the embodiments, "consisting of", refer to the inclusion of exactly one element of a number or list of elements. In general, the term "or" as used herein, when preceded by a term of exclusivity, such as "either", "one of", "only one of", "exactly one of", "consisting essentially of", shall be interpreted only as indicating exclusive alternatives (i.e., "one or the other but not both"), and when used in the embodiments, shall have its ordinary meaning as used in the field of patent law.

[0106] As used herein and in the embodiments, the phrase "at least one" in reference to a list of one or more elements means at least one element selected from any one or more of the elements in the list of elements, but does not necessarily include at least one of each and every element specifically listed in the list of elements, and does not exclude any combination of elements in the list of elements. This definition also allows for the optional presence of elements other than those specifically identified in the list of elements to which the phrase "at least one" refers, whether or not related to the specifically identified elements. Thus, as a non-limiting example, "at least one of A and B" (or, equivalently, "at least one of A or B," or, equivalently, "at least one of A and / or B") can refer, in one embodiment, to at least one, optionally including more than one, A, in which B is absent (and optionally including elements other than B); in another embodiment, to at least one, optionally including more than one, B, in which A is absent (and optionally including elements other than A); in yet another embodiment, to at least one, optionally including more than one, A, and at least one, optionally including more than one, B (and optionally including other elements), etc.

Claims

1. A compound of formula 1, 【Chemical 1】 During the ceremony, A １ is CH ２ , CHR ４ , C.R. ４ R ５ , N.H., N.R. ４ , O or S; A ２ , A ３ , A ４ , and A ５ , CH, CR ６ or N; R １ , R ２ , R ３ , R ４ , R ５ , R ６ is, if present, independently hydrogen, hydroxy, acetyl, halo, carboxyl; substituted or unsubstituted amino, alkyl, alkoxy, carboxyalkyl, alkylamido, acyl, acylaminoaryl, arylalkyl, heteroalkyl, heteroalkoxy, heterocarboxyalkyl, heteroacyl, heteroacylamino, heteroaryl, or heteroarylalkyl; or a substituted or unsubstituted 5-membered heterocycle, where any member of a given ring can be C, N, O, or S. compound.

2. A １ is O, NH, or CH ２ 2. The compound of claim 1, wherein:

3. A ２ The compound of claim 1 or 2, wherein is N.

4. A ３ 3. The compound of claim 1 or 2, wherein is CH or N.

5. A ４ is N or CHCH ３ 3. The compound according to claim 1 or 2, wherein

6. A ５ is CH, CCH ３ , CCF ３ or N.

7. R １ is absent or Cl, F, or CH ３ 3. The compound according to claim 1 or 2, wherein

8. R ２ が、COOH、COOCH ３ 、CONHCH ３ 、CON(CH ３ ) ２ 、 【Chemistry 2】 3. The compound according to claim 1 or 2, wherein

9. R ３ が、COOH、COOCH ２ CH ３ 、CON(CH ３ ) ２ 、CONHCH ２ CH ３ 、 【Chemistry 3】 3. The compound according to claim 1 or 2, wherein

10. Compounds NIM-1001-1 to NIM-1001-39: 【Chemistry 4-1】 【Chemistry 4-2】 【Chemistry 4-3】 【Chemistry 4-4】 【Chemistry 4-5】 【Chemistry 4-6】 2. The compound of claim 1, comprising any one of:

11. A pharmaceutical composition comprising a compound according to claim 1 or 2, wherein the compound is in a pharmaceutically acceptable excipient.

12. 12. The pharmaceutical composition of claim 11, which is capable of preventing or reducing the severity of an inflammatory disease when administered to a mammal suffering from the disease.

13. 13. The pharmaceutical composition of claim 12, wherein the inflammatory disease is systemic inflammation, inflammatory bowel disease, a viral infection that causes inflammation, autoimmune encephalomyelitis, multiple sclerosis, systemic lupus erythematosus (SLE), Alzheimer's disease, Parkinson's disease, psoriasis, or Sjogren's disease.

14. 12. The pharmaceutical composition of claim 11, wherein the inflammatory disease is systemic inflammation and administration of the compound to the mammal reduces spleen size, reduces the percentage of CD4+IL21+ cells in the spleen, and / or increases the percentage of CD25+FOXP3+ cells in the spleen in the mammal.

15. 12. The pharmaceutical composition of claim 11, wherein the inflammatory disease is acute inflammatory bowel disease and administration of the compound reduces the percentage of CD4+ cells in the lamina propria of the colon of the mammal, increases the percentage of CD25+ Treg cells in the colon of the mammal, and / or reduces F4 / 80hi macrophages in the mammal.

16. 12. The pharmaceutical composition of claim 11, wherein the inflammatory disease is a viral infection that causes inflammation, and administration of the compound reduces mortality in a population of mammals infected with the virus.

17. 17. The pharmaceutical composition of claim 16, wherein the virus is influenza A strain H1N1.

18. 12. The pharmaceutical composition of claim 11, wherein the inflammatory disease is autoimmune encephalomyelitis or multiple sclerosis, and administration of the compound reduces the percentage of CD4+IL17+ T cells in the mammal and / or reduces astrocytes or activated glia in the spinal cord of the mammal.

19. 12. The pharmaceutical composition of claim 11, wherein the inflammatory disease is systemic lupus erythematosus (SLE), lupus nephritis, cutaneous lupus, or lupus with central nervous system (CNS) involvement, and the compound reduces urinary protein levels in the mammal, increases CD25+ FOXP3+ regulatory T cells in the mammal, and / or decreases CD4+ IL17+ cells in the mammal.

20. 12. The pharmaceutical composition of claim 11, wherein the inflammatory disease is Alzheimer's disease and administration of the compound reduces anxiety responses by the mammal, reduces neutrophils in the mammal, and / or reduces CD4+IL17+ cells in the mammal.

21. 12. The pharmaceutical composition of claim 11, wherein the inflammatory disease is Parkinson's disease and administration of the compound improves gait in the mammal, reduces CD4+IL21+ cells in the mammal, and / or reduces CD4+IL17+ cells in the mammal.

22. 12. The pharmaceutical composition of claim 11, wherein the inflammatory disease is psoriasis and administration of the compound reduces CD4+TNF+ T cells in the mammal, reduces CD4+IL17+ in the mammal, and / or reduces inflammatory macrophages in the mammal.

23. 12. The pharmaceutical composition of claim 11, wherein the inflammatory disease is Sjogren's syndrome and administration of the compound reduces disease severity and histopathology of the lacrimal and / or salivary glands in a mammal.

24. The pharmaceutical composition of claim 11, wherein the mammal is a human.

25. 12. The pharmaceutical composition of claim 11, formulated for parenteral, enteral, transmucosal, or transdermal administration to a mammal.

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