TLR receptor ligand-based vaccine adjuvant

Abstract

Formula (I) The lipidated oxoadenine of TIFF2022534038000027.tif72149 is a TLR7 / 8 receptor ligand useful for modulating immune responses. The compound may have therapeutic applications in the treatment of cancer, infectious diseases, allergies, or autoimmune disorders.

Description

Technical Field

[0001] Related Applications This application claims priority to U.S. Provisional Patent Application No. 62 / 851,941, filed May 23, 2019, and U.S. Provisional Patent Application No. 62 / 975,054, filed February 11, 2020, the entire contents of which are hereby incorporated by reference.

[0002] Statement Regarding Government Interests This invention was made with government support under Contract No. HHSN272200900036C awarded by the National Institutes of Health. The federal government has certain rights in this invention.

[0003] The present disclosure relates to lipidated TLR7 / 8 receptor ligand compounds and methods of using them, for example, as vaccine adjuvants.

Background Art

[0004] The development of novel vaccine adjuvants and immunotherapies based on Toll-like receptor (TLR) ligands is a rapidly expanding field of research over the past decade, with clinical outcomes across multiple indications and several approved products. Five of the ten known TLRs identified in humans are associated with the recognition of bacterial components (TLR1, 2, 4, 5, 6), and the other four (TLR3, 7, 8, 9) appear to be restricted to intracellular compartments and are involved in the detection of viral RNA (TLR3, 7, 8) and unmethylated DNA (TLR9).

[0005] One of the most promising classes of immunotherapeutic agents includes compounds that target TLR7 / 8. A small number of different classes of small molecules closely resemble the natural (U- and / or G-rich) viral ssRNA ligands of TLR7 / 8. These include oxoguanosines that interact primarily with TLR7 and derivatives of adenine that bind to TLR7 and / or TLR8. One class of TLR-active adenine derivatives is oxoadenines, which were originally developed to overcome certain side effects associated with imidazoquinolines. Despite the fact that the oxoadenine class exhibits an overall toxicity / biological activity profile superior to that of imidazoquinolines, they can still lead to systemic inflammatory responses upon administration, limiting their use in the human medical setting. In fact, most TLR7 / 8 agonists currently under development tend to be toxic, insoluble or unstable, and / or have very low immunostimulatory effects.

Summary of the Invention

Problems to be Solved by the Invention

[0006] Therefore, there is a need for effective and safe TLR7 and / or TLR8 receptor ligand compounds for vaccines and immunotherapeutic agents.

Means for Solving the Problems

[0007] In one aspect, formula (I):

Chemical formula

[0008] In another aspect, the present invention provides a pharmaceutical composition comprising a compound of formula (I) or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier.

[0009] In another aspect, the present invention provides a vaccine composition comprising a compound of formula (I) or a pharmaceutically acceptable salt thereof and an antigen.

[0010] In another aspect, the present invention provides a formulation comprising microparticles or nanoparticles comprising a compound of formula (I) or a pharmaceutically acceptable salt thereof.

[0011] In another aspect, the present invention provides an adjuvant composition comprising a compound of formula (I) or a pharmaceutically acceptable salt thereof.

[0012] In another aspect, the present invention provides a method of modulating an immune response in a subject, the method comprising administering to the subject a therapeutically effective amount of a compound of formula (I) or a pharmaceutically acceptable salt thereof or a pharmaceutical composition thereof.

[0013] Another aspect of the present invention is a method for treating, preventing, or reducing susceptibility to cancer in a subject, which comprises administering to the subject in need thereof a therapeutically effective amount of a compound of formula (I), a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof.

[0014] Another aspect of the present invention is a method for treating, preventing, or reducing susceptibility to an infectious disease in a subject, which comprises administering to the subject in need thereof a therapeutically effective amount of a compound of formula (I), a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof.

[0015] Another aspect of the present invention is a method for treating, preventing, or reducing susceptibility to allergy in a subject, which comprises administering to the subject in need thereof a therapeutically effective amount of a compound of formula (I), a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof.

[0016] Another aspect of the present invention is a method for treating, preventing, or reducing susceptibility to an autoimmune disease in a subject, which comprises administering to the subject in need thereof a therapeutically effective amount of a compound of formula (I), a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof.

[0017] In another aspect, the present invention provides a compound of formula (I), a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof for use in a method of modulating an immune response.

[0018] In another aspect, the present invention provides a compound of formula (I), a pharmaceutically acceptable salt thereof, or a pharmaceutical composition thereof for use in a method for treating, preventing, or reducing susceptibility to cancer, an infectious disease, an allergy, or an autoimmune disease.

[0019] In another aspect, the present invention provides the use of a compound of formula (I) or a pharmaceutically acceptable salt thereof or a pharmaceutical composition thereof for the manufacture of a medicament for modulating an immune response.

[0020] In another aspect, the present invention provides the use of a compound of formula (I) or a pharmaceutically acceptable salt thereof or a pharmaceutical composition thereof for the manufacture of a medicament for treating, preventing, or reducing susceptibility to cancer, infectious diseases, allergies, or autoimmune diseases.

[0021] Other aspects and embodiments of the present disclosure will become apparent in light of the following description and drawings.

Brief Description of the Drawings

[0022]

Figure 1

Figure 2

Figure 3-1

Figure 3-2

Figure 4

Figure 5-1

Figure 5-2

Figure 6

Mode for Carrying Out the Invention

[0023] Classes of compounds are described herein that can act as TLR7 / 8 ligands and may be useful as novel vaccine adjuvants and in immunotherapy. The TLR7 / 8 ligands are novel pegylated and lipidated oxoadenine compounds. The compounds of the invention have been shown to be inducers of interferon-α and other immunostimulatory cytokines and may have an improved activity-toxicity profile compared to other known oxoadenine-based TLR7 / 8 ligands.

[0024] 1. Definitions Unless otherwise defined, all scientific and technical terms used herein have the same meaning as commonly understood by one of ordinary skill in the art. In case of conflict, the present specification, including definitions, will control. Preferred methods and materials are described below, but methods and materials similar or equivalent to those described herein can be used in the practice or testing. All publications, patent applications, patents, and other references mentioned herein are incorporated by reference in their entirety. The materials, methods, and examples disclosed herein are illustrative only and not intended to be limiting.

[0025] The terms "comprising," "including," "having," "has," "can," "containing," and variations thereof are intended as open-ended transitional phrases, terms, or statements that do not exclude the possibility of additional acts or structures when used herein. The singular forms "a," "an," and "the" include references to the plural unless the context clearly dictates otherwise. The present disclosure contemplates other embodiments that "comprise," "consist of," and "consist essentially of" the embodiments or elements provided herein, whether explicitly stated or not.

[0026] The modifier "about" as used in connection with a quantity includes the recited value and has a meaning determined by the context (e.g., this includes at least the degree of error associated with the measurement of a particular quantity). The modifier "about" must also be considered to disclose a range defined by the absolute values of two endpoints. For example, the expression "about 2 to about 4" also discloses the range "2 to 4". The term "about" can refer to ±10% of the indicated number. For example, "about 10%" can indicate a range of 9% to 11%, and "about 1" can mean 0.9 to 1.1. Other meanings of "about" can be apparent from the context, such as rounding, and thus, for example, "about 1" can also mean 0.5 to 1.4.

[0027] The term "immune response" includes, but is not limited to, any response related to innate and adaptive immunity, including cytokine expression, production or secretion (e.g., IL-1, IL-6, IL-17, TNFα expression, production or secretion), cytotoxicity, immune cell migration, antibody production and / or an increase or decrease in immune cell response.

[0028] The phrase "modulating an immune response" or "modulation of an immune response" or "modulates an immune response" includes upregulating, enhancing, stimulating, augmenting or increasing an immune response as defined herein.

[0029] The definitions of specific functional groups and chemical terms are described in further detail below. For the purposes of the present disclosure, chemical elements are identified according to the CAS version of the Periodic Table of the Elements, Handbook of Chemistry and Physics, 75 th Ed. on the inside front cover, and specific functional groups are generally defined as described therein. In addition, for general principles of organic chemistry as well as specific functional moieties and reactivity, see Organic Chemistry, Thomas Sorrell, University Science Books, Sausalito, 1999; Smith and March March’s Advanced Organic Chemistry, 5th Edition, John Wiley & Sons, Inc., New York, 2001; Larock, Comprehensive Organic Transformations, VCH Publishers, Inc., New York, 1989; Carruthers, Some Modern Methods of Organic Synthesis, 3 rd rd Edition, Cambridge University Press, Cambridge, 1987, the entire contents of each of which are incorporated herein by reference.

[0030] As used herein, the term "alkyl" means a straight or branched chain saturated hydrocarbon chain containing 1 to 10 carbon atoms. The term "lower alkyl" or "C 1 ~C 6 -alkyl" means a straight or branched hydrocarbon containing 1 to 6 carbon atoms. The term "C 1 ~C 3 -alkyl" means a straight or branched hydrocarbon containing 1 to 3 carbon atoms. Representative examples of alkyl include, but are not limited to, methyl, ethyl, n-propyl, isopropyl, n-butyl, sec-butyl, isobutyl, tert-butyl, n-pentyl, isopentyl, neopentyl, n-hexyl, 3-methylhexyl, 2,2-dimethylpentyl, 2,3-dimethylpentyl, 4,4-dimethylpentan-2-yl, n-heptyl, n-octyl, n-nonyl and n-decyl.

[0031] As used herein, the term "alkenyl" means a straight or branched chain hydrocarbon chain containing at least one carbon-carbon double bond and 1 to 10 carbon atoms.

[0032] As used herein, the term "alkylene" refers to a divalent group derived from a straight or branched hydrocarbon having 1 to 10 carbon atoms, such as 2 to 5 carbon atoms. Representative examples of alkylene include, but are not limited to, -CH 2 CH2 -, -CH 2 CH 2 CH 2 -, -CH 2 CH(CH 3 )CH 2 -, -CH 2 CH 2 CH 2 CH 2 -, -CH 2 CH(CH 3 )CH 2 CH 2 - and -CH 2 CH 2 CH 2 CH 2 CH 2 - are included.

[0033] In some examples, the number of carbon atoms in a hydrocarbyl substituent (e.g., alkyl or cycloalkyl) is indicated by the prefix "C x ~C y -", where x is the minimum number of carbon atoms in the substituent and y is the maximum number. Thus, for example, "C 1 ~C 3 -alkyl" refers to an alkyl substituent containing 1 to 3 carbon atoms.

[0034] The term "substituent" refers to a group that is "substituted" at any atom of that group on an aryl, heteroaryl, phenyl, or pyridinyl group. Any atom can be substituted.

[0035] The term "substituted" refers to a group that may be further substituted with one or more non-hydrogen substituents. Substituents include, but are not limited to, halogen, =O (oxo), =S (thioxo), cyano, nitro, fluoroalkyl, alkoxyfluoroalkyl, fluoroalkoxy, alkyl, alkenyl, alkynyl, haloalkyl, haloalkoxy, heteroalkyl, cycloalkyl, cycloalkenyl, aryl, heteroaryl, heterocycle, cycloalkylalkyl, heteroarylalkyl, arylalkyl, hydroxy, hydroxyalkyl, alkoxy, alkoxyalkyl, alkylene, aryloxy, phenoxy, benzyloxy, amino, alkylamino, acylamino, aminoalkyl, arylamino, sulfonylamino, sulfinylamino, sulfonyl, alkylsulfonyl, arylsulfonyl, aminosulfonyl, sulfinyl, -COOH, ketone, amide, carbamate and acyl. For example, when a group is described as "optionally substituted" (such as other groups like alkyl, alkenyl, alkynyl, aryl, heteroaryl, cycloalkyl, heteroalkyl, heterocycle or R group), it may have 0, 1, 2, 3, 4 or 5 substituents independently selected from halogen, =O (oxo), =S (thioxo), cyano, nitro, fluoroalkyl, alkoxyfluoroalkyl, fluoroalkoxy, alkyl, alkenyl, alkynyl, haloalkyl, haloalkoxy, heteroalkyl, cycloalkyl, cycloalkenyl, aryl, heteroaryl, heterocycle, cycloalkylalkyl, heteroarylalkyl, arylalkyl, hydroxy, hydroxyalkyl, alkoxy, alkoxyalkyl, alkylene, aryloxy, phenoxy, benzyloxy, amino, alkylamino, acylamino, aminoalkyl, arylamino, sulfonylamino, sulfinylamino, sulfonyl, alkylsulfonyl, arylsulfonyl, aminosulfonyl, sulfinyl, -COOH, ketone, amide, carbamate and acyl.

[0036] The term

Chem.

[0037] For the compounds described herein, the groups and their substituents can be selected according to the acceptable valences of the atoms and substituents such that the selection and substitution do not spontaneously cause conversions such as rearrangement, cyclization, elimination, etc., which result in stable compounds.

[0038] When a substituent is specified by its conventional chemical formula written from left to right, such a formula also includes the same substituent that can result from writing the structure from right to left. For example, -CH 2 NH- means -NHCH 2 - is also intended to be included.

[0039] Regarding the description of numerical ranges herein, each intervening number between them with the same precision is explicitly contemplated. For example, for the range of 6 to 9, in addition to 6 and 9, the numbers 7 and 8 are contemplated, and for the range of 6.0 to 7.0, the numbers 6.0, 6.1, 6.2, 6.3, 6.4, 6.5, 6.6, 6.7, 6.8, 6.9, and 7.0 are explicitly contemplated.

[0040] 2. Compounds In one aspect, formula (I):

Chemical formula

[0041] In some embodiments, R 2 is hydrogen or C(O)R 4 , where R 4 , when present, in each occurrence, is (CH 2 ) 10 CH 3 , (CH 2 ) 12 CH 3 、(CH 2 ) 14 CH 3 、(CH 2 ) 16 CH 3 and (CH 2 ) 7 CH = CH(CH 2 ) 7 CH 3 independently selected from.

[0042] In some embodiments, R 3 is C(O)R 4 , where R 4 , in each occurrence, is (CH 2 ) 10 CH 3 , (CH 2 ) 12 CH 3 , (CH 2 ) 14CH 3 and (CH 2 ) 16 CH 3 and (CH 2 ) 7 CH=CH(CH 2 ) 7 CH 3 independently selected from.

[0043] In some embodiments, R 4 is, in each occurrence, (CH 2 ) 10 CH 3 , (CH 2 ) 12 CH 3 , (CH 2 ) 14 CH 3 , (CH 2 ) 16 CH 3 and (CH 2 ) 7 CH=CH(CH 2 ) 7 CH 3 independently selected from. In some embodiments, R 4 is (CH 2 ) 14 CH 3 .

[0044] In some embodiments, R 2 and R 3 are each C(O)R 4 , where R 4 is, in each occurrence, (CH 2 ) 10 CH 3 , (CH 2 ) 12 CH 3 , (CH 2 ) 14 CH 3 , (CH 2 ) 16 CH 3 and (CH 2 ) 7 CH=CH(CH 2 ) 7 CH 3is independently selected from. In some embodiments, R 2 and R 3 are each C(O)R 4 wherein R 4 is, in each occurrence, (CH 2 ) 14 CH 3 .

[0045] In some embodiments, R 2 is hydrogen and R 3 is C(O)R 4 wherein R 4 is (CH 2 ) 10 CH 3 , (CH 2 ) 12 CH 3 , (CH 2 ) 14 CH 3 , (CH 2 ) 16 CH 3 and (CH 2 ) 7 CH=CH(CH 2 ) 7 CH 3 selected from.

[0046] In some embodiments, n is 1. In some embodiments, m is 2. In an exemplary embodiment, n is 1 and m is 2.

[0047] In some embodiments, Z is (C 2 alkylene - O) q wherein q is 3, 6, 9, 12 or 16. In an exemplary embodiment, q is 3. q can be 3, 6, 9, 12 or 16 in any embodiment herein.

[0048] The alkylene of Z can be a straight - chain alkylene, i.e., n - alkylene. For example, Z can be (C 2 ~C 6 n - alkylene - O) q . Z is (CH2 CH 2 -O) q may be. Z is (CH 2 CH 2 -O) 3 may be. Z is (CH 2 CH 2 -O) 6 may be, or Z is (CH 2 CH 2 -O) 9 may be, or Z is (CH 2 CH 2 -O) 12 may be, or Z is (CH 2 CH 2 -O) 16 may be.

[0049] In the compounds disclosed herein, Z is directed by the terminal carbon of Z attached to the phosphate moiety and the oxygen of Z that is part of the glycerol moiety, as shown in formula (II).

Chemical formula

[0050] In another aspect, formula (Ia):

Chemical formula

[0051] In some embodiments, R 2 is hydrogen or C(O)R 4 , where R 4 , when present, in each occurrence, is (CH 2 ) 10 CH 3 , (CH 2 ) 12 CH 3 , (CH 2 ) 14 CH 3 , (CH 2 ) 16 CH 3 and (CH 2 ) 7 CH = CH(CH 2 ) 7 CH 3 independently selected from.

[0052] In some embodiments, R 3 is C(O)R 4 , where R 4 , in each occurrence, is (CH 2 ) 10 CH 3 , (CH 2 ) 12 CH 3 , (CH 2 ) 14 CH 3 、(CH2 ) 16 CH 3 and (CH 2 ) 7 CH=CH(CH 2 ) 7 CH 3 is independently selected from.

[0053] In some embodiments, R 4 is, in each occurrence, (CH 2 ) 10 CH 3 , (CH 2 ) 12 CH 3 , (CH 2 ) 14 CH 3 , (CH 2 ) 16 CH 3 and (CH 2 ) 7 CH=CH(CH 2 ) 7 CH 3 is independently selected from. In some embodiments, R 4 is (CH 2 ) 14 CH 3 .

[0054] In some embodiments, R 2 and R 3 are each C(O)R 4 , where R 4 is, in each occurrence, (CH 2 ) 10 CH 3 , (CH 2 ) 12 CH 3 , (CH 2 ) 14 CH 3 , (CH 2 ) 16 CH 3 and (CH 2 ) 7 CH=CH(CH 2 ) 7 CH 3 is independently selected from. In some embodiments, R 2and R 3 is each C(O)R 4 wherein R 4 is, in each occurrence, (CH 2 ) 14 CH 3 as defined herein.

[0055] In some embodiments, R 2 is hydrogen and R 3 is C(O)R 4 wherein R 4 is (CH 2 ) 10 CH 3 , (CH 2 ) 12 CH 3 , (CH 2 ) 14 CH 3 , (CH 2 ) 16 CH 3 and (CH 2 ) 7 CH=CH(CH 2 ) 7 CH 3 selected from.

[0056] In some embodiments, n is 1. In some embodiments, m is 2. In an exemplary embodiment, n is 1 and m is 2.

[0057] In some embodiments, Z is (C 2 alkylene - O) q wherein q is 3, 6, 9, 12 or 16. In an exemplary embodiment, q is 3.

[0058] The compound or a pharmaceutically acceptable salt thereof is

Chemical formula

Chemical formula

Chemical formula

[0059] The compound may exist as stereoisomers having an asymmetric center or a chiral center. The stereoisomers are "R" or "S" depending on the configuration of the substituents around the chiral carbon atom. The terms "R" and "S" as used herein are the configurations as defined in IUPAC 1974 Recommendations for Section E, Fundamental Stereochemistry, in Pure Appl. Chem., 1976, 45:13 - 30. The present disclosure contemplates various stereoisomers and mixtures thereof, which are specifically included within the scope of the present disclosure. Stereoisomers include enantiomers and diastereomers and mixtures of enantiomers or diastereomers. The individual stereoisomers of the compound can be prepared by synthesis from commercially available starting materials containing an asymmetric center or a chiral center, or by the preparation of a racemic mixture followed by a resolution method well known to those skilled in the art. Such resolution methods include (1) the attachment of an enantiomeric mixture to a chiral auxiliary as described in Furniss, Hannaford, Smith, and Tatchell, "Vogel’s Textbook of Practical Organic Chemistry," 5th edition (1989), Longman Scientific & Technical, Essex CM20 2JE, England (or its latest edition), the recrystallization or chromatographic separation of the resulting mixture of diastereomers and the optional liberation of the optically pure product from the auxiliary, or (2) the direct separation of an optically active enantiomeric mixture on a chiral chromatography column, or (3) exemplified by the fractional recrystallization method.

[0060] It should be understood that the compound may have tautomers and geometric isomers, and these also constitute embodiments of the present disclosure.

[0061] The present disclosure also includes compounds labeled with isotopes, which are generally identical to those described in formula (I), except that one or more atoms are replaced by atoms having an atomic mass or mass number different from the atomic mass or mass number normally found in nature. Examples of isotopes suitable for inclusion in the compounds of the present disclosure include, but are not limited to, 2 H, 3 H, 13 C, 14 C, 15 N, 18 O, 17 O, 31 P, 32 P, 35 S, 18 F and 36 Cl, etc., of hydrogen, carbon, nitrogen, oxygen, phosphorus, sulfur, fluorine and chlorine. Substitution with heavier isotopes such as deuterium, i.e., 2 H, etc., can result in certain therapeutic advantages obtained from higher metabolic stability, such as an increase in in vivo half-life or a decrease in the required dosage, and is therefore sometimes preferred depending on the situation. The compounds can incorporate positron-emitting isotopes for medical imaging and positron emission tomography (PET) studies to determine receptor distribution. Suitable positron-emitting isotopes that can be incorporated into the compounds of formula (I) are 11 C, 13 N, 15 O and 18 F. Compounds of formula (I) labeled with isotopes can generally be prepared by methods similar to those known to those skilled in the art or described in the attached examples, using appropriately labeled reagents in place of unlabeled reagents.

[0062] a. pharmaceutically acceptable salts The disclosed compounds can exist as pharmaceutically acceptable salts. The term "pharmaceutically acceptable salts" refers to salts or zwitterions of compounds that are soluble or dispersible in water or oil, suitable for the treatment of disorders, free of excessive toxicity, irritation, and allergic reactions, commensurate with a reasonable risk-benefit ratio, and effective for their intended use. The salts can be prepared during the final isolation and purification of the compound or separately by reacting the amino groups of the compound with a suitable acid. For example, without limitation, the compound can be dissolved in a suitable solvent such as methanol and water and treated with at least one equivalent of an acid such as hydrochloric acid. The resulting salt can be precipitated, separated by filtration, and dried under reduced pressure. Alternatively, the solvent and the excess amount of acid can be removed under reduced pressure to provide the salt. Representative salts include acetate, adipate, alginate, citrate, aspartate, benzoate, benzenesulfonate, bisulfate, butyrate, camphorate, camphorsulfonate, digluconate, glycerophosphate, hemisulfate, heptanoate, hexanoate, formate, isethionate, fumarate, lactate, maleate, methanesulfonate, naphthalenesulfonate, nicotinate, oxalate, pamoate, pectinate, persulfate, 3-phenylpropionate, picrate, oxalate, maleate, pivalate, propionate, succinate, tartrate, trichloroacetate, trifluoroacetate, glutamate, paratoluenesulfonate, undecanoate, hydrochloride, hydrobromide, sulfate, phosphate, etc. The amino groups of the compound can also be quaternized with alkyl chlorides, bromides, and iodides such as methyl, ethyl, propyl, isopropyl, butyl, lauryl, myristyl, stearyl, etc.

[0063] Basic addition salts can be prepared by reaction of the carboxyl or phosphate groups with a suitable base, such as hydroxides, carbonates or bicarbonates of metal cations such as lithium, sodium, potassium, calcium, magnesium or aluminum, or organic primary, secondary or tertiary amines, during the final separation and purification of the disclosed compounds. Quaternary amine salts can be prepared, such as those derived from methylamine, dimethylamine, trimethylamine, triethylamine, diethylamine, ethylamine, tributylamine, pyridine, N,N-dimethylaniline, N-methylpiperidine, N-methylmorpholine, dicyclohexylamine, procaine, dibenzylamine, N,N-dibenzylphenethylamine, 1-ephenamine and N,N'-dibenzylethylenediamine, ethylenediamine, ethanolamine, diethanolamine, piperidine, piperazine and the like. In some embodiments, the pharmaceutically acceptable salt is a choline salt.

[0064] Compounds and intermediates can be separated and purified by methods well known to those skilled in the art of organic synthesis. Examples of conventional methods for separating and purifying compounds include, but are not limited to, chromatography on solid supports such as silica gel, alumina or silica derivatized with alkylsilane groups, recrystallization at high or low temperature with optional pretreatment with activated carbon, thin layer chromatography, distillation at various pressures, sublimation under vacuum and trituration, as described in, for example, “Vogel’s Textbook of Practical Organic Chemistry,” 5th edition (1989), by Furniss, Hannaford, Smith, and Tatchell, pub. Longman Scientific & Technical, Essex CM20 2JE, England.

[0065] The compounds described herein or their pharmaceutically acceptable salts may be included in formulations. The formulations may include microparticles or nanoparticles including, but not limited to, liposomes, micelles, polymers, block copolymers, silica, emulsions, or combinations thereof. The formulations may include organic, inorganic, and / or lipid constituents, may contribute to the immunomodulatory effects of the compounds, and / or may affect properties such as the biodistribution, bioavailability, and / or toxicity of the compounds.

[0066] b. Biological activity The compounds disclosed herein, including the compounds of formula (I), may have biological activities that render them useful as immunological adjuvants or immunomodulatory agents. For example, the compounds may stimulate the immune system's response to a co-administered antigen. In some embodiments, the compounds of formula (I) are TLR7 antagonists. In some embodiments, the compounds of formula (I) are TLR8 antagonists. In some embodiments, the compounds have activity as Th1-stimulating adjuvants.

[0067] In some embodiments, the compounds may stimulate the production of cytokines in a sample or upon administration to a subject. The compounds may stimulate the production of Th17-type cytokines. Exemplary cytokines include IFN-γ, IL-2, and IL-12. Such activities may be tested by established methods. For example, the levels of such cytokines may be measured in samples of peripheral blood mononuclear cells (PBMC) after exposure to the compounds.

[0068] Conditions that may be mediated by the activity of TLR7 and / or TLR8 include, but are not limited to, inflammation such as inflammatory or allergic diseases such as asthma, allergic rhinitis, hypersensitivity pneumonitis, reflux syndrome, delayed hypersensitivity, atherosclerosis, pancreatitis, gastritis, osteoarthritis, psoriasis, sarcoidosis, pulmonary fibrosis, respiratory distress syndrome, bronchiolitis, chronic obstructive pulmonary disease, rhinosinusitis, cystic fibrosis and dermatitis; autoimmune diseases including, but not limited to, rheumatoid arthritis, psoriatic arthritis, systemic lupus erythematosus, Sjogren's syndrome, ankylosing spondylitis, scleroderma, diabetes, graft rejection including graft-versus-host disease, and inflammatory bowel diseases including, but not limited to, Crohn's disease and ulcerative colitis; infectious diseases including, but not limited to, those caused by hepatitis viruses (e.g., hepatitis B virus, hepatitis C virus), human immunodeficiency virus, papillomavirus, herpes virus, respiratory viruses (e.g., influenza virus, respiratory syncytial virus, rhinovirus, metapneumovirus, parainfluenza virus, SARS) and West Nile virus; microbial infections caused by bacteria, fungi or protozoa including, but not limited to, tuberculosis, bacterial pneumonia, aspergillosis, histoplasmosis, candidiasis, pneumocystis, leprosy, chlamydia, cryptococcosis, cryptosporidiosis, toxoplasmosis, leishmania, malaria and trypanosomiasis; the treatment of cancers known to respond to immunotherapy including, but not limited to, various cancers such as renal cell carcinoma, lung cancer, breast cancer, colorectal cancer, bladder cancer, melanoma, leukemia, lymphoma and ovarian cancer; basal cell carcinoma; actinic keratosis; genital papillomavirus infection; and liver regeneration. The treatment of cancers known to respond to immunotherapy including, but not limited to, various cancers such as renal cell carcinoma, lung cancer, breast cancer, colorectal cancer, bladder cancer, melanoma, leukemia, lymphoma and ovarian cancer; basal cell carcinoma; actinic keratosis; genital papillomavirus infection; and liver regeneration are included, but not limited to these.

[0069] 3. Composition The disclosed compounds can be incorporated into pharmaceutical compositions, adjuvant compositions, and vaccine compositions that may be suitable for administration to a subject (such as a patient, which can be human or non - human).

[0070] a. Pharmaceutical compositions The disclosed compounds can be incorporated into pharmaceutical compositions. The pharmaceutical compositions can contain a "therapeutically effective amount" or a "prophylactically effective amount" of the agent. A "therapeutically effective amount" refers to an amount effective to achieve the desired therapeutic result at the required dosage and for the required period. The therapeutically effective amount of the composition can be determined by one of ordinary skill in the art and can vary depending on factors such as the individual's disease state, age, gender, and weight, as well as the ability of the composition to elicit the desired response in the individual. The therapeutically effective amount is also one in which any toxic or adverse effects of the compounds of the present invention (e.g., compounds of formula (I)) are outweighed by the therapeutically beneficial effects. A "prophylactically effective amount" refers to an amount effective to achieve the desired prophylactic result at the required dosage and for the required period. Typically, prophylactic dosages are used prior to or at the initial stages of a disease in a subject, and thus the prophylactically effective amount will be less than the therapeutically effective amount.

[0071] Pharmaceutical compositions and formulations can further contain additional therapeutic agents. In some embodiments, the additional therapeutic agents are adjuvants, immunostimulants, chemotherapeutic agents, immunomodulators, or combinations thereof.

[0072] Pharmaceutical compositions and formulations can contain an adjuvant. An adjuvant is an additive that enhances the humoral and / or cell - mediated immune response to a vaccine antigen. Any adjuvant can be useful in the pharmaceutical compositions and formulations described herein. An adjuvant can interact with one member of the TLR family of members. In some embodiments, the adjuvant is a TLR4 ligand.

[0073] TLR4 ligands are CRX - 601 (Figure 1), monophosphoryl lipid A (MPLA), glucopyranosyl lipid A (GLA), CRX - 547,

Chemical formula

Chemical formula

Chemical formula

Chemical formula

[0074] In some embodiments, the adjuvant is an aluminum salt. The aluminum salt can include phosphoric acid, sulfuric acid, hydroxide, or a combination thereof. In some embodiments, the aluminum salt is potassium aluminum sulfate, which can be in the form of a hydrate. The aluminum salt can be aluminum hydroxide gel. The aluminum salt can be aluminum phosphate wet gel. The aluminum salt adjuvant can be referred to as alum. In some embodiments, the compounds disclosed herein are adsorbed to the aluminum salt.

[0075] The pharmaceutical composition may further comprise an antigen. In some embodiments, the antigen is adsorbed to an aluminum salt together with the compounds disclosed herein. Suitable antigens include microbial pathogens, bacteria, viruses, proteins, glycoproteins, lipoproteins, peptides, glycopeptides, lipopeptides, toxoids, carbohydrates, and tumor-specific antigens. A mixture of two or more antigens may be used. In some embodiments, the antigen is derived from bacteria, viruses, bacteriophages, fungi, prions, neoplasms, autoantigens, animals, plants, recombinant or synthetic substances. In one embodiment, the antigen in the vaccine composition is in the form of a peptide, polypeptide, protein, or an immunogenic portion thereof. In some embodiments, the antigen is a hapten, a hapten conjugated to a carrier protein, a polypeptide, or other polymer or a derivative thereof. In some embodiments, the antigen is an allergen.

[0076] The pharmaceutical compositions and formulations may include an immunostimulant. An immunostimulant stimulates the immune system by inducing the activation or increasing the activity of any component of the immune system. Any of a number of known molecules or compounds having activity as an immunostimulant can be used in the disclosed pharmaceutical compositions and formulations.

[0077] The pharmaceutical composition and the preparation may contain a chemotherapeutic agent. The chemotherapeutic agent may include any drug or any radiosensitizer used in cancer treatment. The chemotherapeutic agent may include alkylating agents (including, but not limited to, cyclophosphamide, mechlorethamine, chlorambucil, melphalan, dacarbazine, nitrosourea, and temozolomide), anthracyclines (including, but not limited to, daunorubicin, doxorubicin, epirubicin, idarubicin, mitoxantrone, and valrubicin), cytoskeletal disrupting agents or taxanes (including, but not limited to, paclitaxel, docetaxel, abraxane, and taxotere), epothilones, histone deacetylase inhibitors (including, but not limited to, vorinostat and romidepsin), topoisomerase inhibitors (including, but not limited to, irinotecan, topotecan, etoposide, teniposide, and talaporfin), kinase inhibitors (including, but not limited to, bortezomib, erlotinib, gefitinib, imatinib, vemurafenib, and vismodegib), nucleotide analogs and precursor analogs (including, but not limited to, azacitidine, azathioprine, capecitabine, cytarabine, doxifluridine, fluorouracil, gemcitabine, hydroxyurea, mercaptopurine, methotrexate, and thioguanine), peptide antibiotics (including, but not limited to, bleomycin and actinomycin), platinum-based agents (including, but not limited to, carboplatin, cisplatin, and oxaliplatin), retinoids (including, but not limited to, tretinoin, alitretinoin, and bexarotene), vinca alkaloids and derivatives (including, but not limited to, vinblastine, vincristine, vindesine, and vinorelbine), or a combination thereof.

[0078] The pharmaceutical composition and the preparation may contain an immunomodulator. The immunomodulator includes interferons, antigens, tumor phagocytosis inducers, and other immunopotentiators (e.g., immune checkpoint inhibitors). The immunomodulator may be an immune checkpoint inhibitor, a tumor phagocytosis inducer, or a combination thereof.

[0079] Interferons include interferon alpha, interferon alpha-2a, interferon alpha-2b, interferon beta, interferon gamma-la, ACTIMMUNE® (interferon gamma-lb) or interferon gamma-nl, combinations and homologues thereof.

[0080] In some embodiments, the immunomodulatory agent is an immune checkpoint inhibitor. Immune checkpoints regulate the function of T cells in the immune system. T cells play a central role in cell-mediated immunity. Checkpoint proteins send signals to T cells to interact with specific ligands that essentially switch off or inhibit the function of T cells. Therapy with checkpoint inhibitors "removes the obstacles" to an existing immune response or removes what is preventing the initiation of an immune response. Since many immune checkpoints are regulated by the interaction between specific receptor-ligand pairs, monoclonal antibodies or other agents can be used to block this interaction and prevent immunosuppressive effects. Checkpoint inhibitors can be biological therapeutics, small molecules, monoclonal antibodies, humanized antibodies, fully human antibodies, fusion proteins or combinations thereof. Checkpoint inhibitors can inhibit checkpoint proteins including, for example, CTLA-4, PDL1, PDL2, PD1, B7-H3, B7-H4, BTLA, HVEM, TIM3, GAL9, LAG3, VISTA, KIR, 2B4, CD160, CGEN-15049, CHK1, CHK2, A2aR and B-7 family ligands. Checkpoint inhibitors can interact with ligands of checkpoint proteins including, for example, CTLA-4, PDL1, PDL2, PD1, B7-H3, B7-H4, BTLA, HVEM, TIM3, GAL9, LAG3, VISTA, KIR, 2B4, CD160, CGEN-15049, CHK1, CHK2, A2aR and B-7 family ligands.

[0081] Immune checkpoint inhibitors include PD-1 inhibitors (e.g., nivolumab, pidilizumab, sintilimab), PD-L1 inhibitors (e.g., atezolizumab, avelumab, durvalumab, BMS-936559), CTLA4 inhibitors (e.g., ipilimumab, tremelimumab), or IDO inhibitors (e.g., indoximod, epacadostat).

[0082] In some embodiments, the immunomodulator is a tumor phagocytosis inducer. Immune-stimulatory cell surface polypeptides and their receptors are important for the elimination and destruction of foreign substances including mammalian cells or bacteria. Immune-stimulatory cell surface polypeptides and their receptors activate phagocytosis. Phagocytosis inducers can induce cell surface polypeptides and their receptors to activate phagocytosis. Tumor phagocytosis inducers include anti-CD47 monoclonal antibodies (e.g., Hu5F9-G4, CC-90002, ZF1, AMMS4-G4, IBI188, SRF231), anti-SIRPα fusion proteins (e.g., TTI-621, TTI-622), anti-SIRPα monoclonal antibodies (e.g., OSE-172), anti-CD47 / anti-tumor associated antigen bispecific antibodies, and inhibitors of leukocyte immunoglobulin-like receptor B1 (LILRB1) that binds to major histocompatibility complex class 1 β2-microglobulin (MHC class 1 β2M). Anti-CD47 / anti-tumor associated antigen bispecific antibodies include anti-CD47 / CD19 bispecific antibodies (e.g., TG-1801), anti-CD47 / mesothelin bispecific antibodies (e.g., NI-1801), anti-CD47 / 4-1BB bispecific antibodies (e.g., DSP107), anti-CD47 / CD20 bispecific antibodies, anti-CD47 / CD33 bispecific antibodies (e.g., HMBD004).

[0083] Other immune-regulating agents include, but are not limited to, the same kind of agents including ALFAFERONE (registered trademark), BAM-002, BEROMUN (registered trademark) (tasonermin), BEXXAR (registered trademark) (tositumomab), CamPath (registered trademark) (alemtuzumab), CTLA4 (cytotoxic lymphocyte antigen 4), dacarbazine, denileukin, epratuzumab, GRANOCYTE (registered trademark) (lenograstim), lentinan, leukocyte alpha interferon, imiquimod, MDX-010, melanoma vaccine, mitsumomab, molgramostim, MYLOTARGTM (registered trademark) (gemtuzumab ozogamicin), NEUPOGEN (registered trademark) (filgrastim), OncoVAC-CL, OvaRex (registered trademark) (oregovomab), pemtumomab (Y-muHMFGl), PROVENGE (registered trademark), sargaramostim, schizophyllan, tasonermin, TheraCys (registered trademark), ubenimex, VIRULIZIN (registered trademark), Z-100, WF-10, PROLEUKIN (registered trademark) (aldesleukin), ZADAXIN (registered trademark) (thymalfasin), ZENAPAX (registered trademark) (daclizumab), ZEVALIN (registered trademark) (90Y-ibritumomab tiuxetan), and STING (stimulator of interferon genes) and NOD (nucleotide-binding oligomerization domain-like receptor) agonists.

[0084] In the methods and uses described herein, the pharmaceutical combination of a compound of formula (I) or a pharmaceutically acceptable salt or composition thereof and an adjuvant and / or immunomodulatory agent can be administered / used simultaneously, separately or sequentially, in any order, and the components can be administered separately or as a fixed combination. For example, a therapeutic treatment according to the invention can comprise the administration of a therapeutically effective amount or an effective amount, for example a daily dosage corresponding to the amounts described herein, of a first active ingredient in free form or in the form of a pharmaceutically acceptable salt and of a second active ingredient in free form or in the form of a pharmaceutically acceptable salt, simultaneously or sequentially, in any order. The individual active ingredients of the combination can be administered separately or in parallel at different times during the course of the therapy, in a divided or single dosage form. The present invention should therefore be understood as encompassing all such treatment programs, such as simultaneous or alternating therapies, and the term "administering" should be construed accordingly. Thus, a pharmaceutical combination, as used herein, defines a fixed combination for combined administration in one dosage unit form or in separate dosage forms, and the combined administration can be carried out independently, simultaneously or at different times. As a further example, an adjuvant or immunomodulatory agent can be administered / used simultaneously (e.g., by co-infusion), separately or sequentially, followed by the administration of a compound of formula (I) or vice versa.

[0085] The pharmaceutical compositions and formulations may contain a pharmaceutically acceptable carrier. As used herein, the term "pharmaceutically acceptable carrier" means any kind of non-toxic, inert solid, semi-solid or liquid filler, diluent, encapsulating material or formulation aid. Some examples of materials that can serve as pharmaceutically acceptable carriers include, but are not limited to, saccharides such as lactose, glucose and sucrose; starches such as corn starch and potato starch; celluloses and their derivatives such as sodium carboxymethyl cellulose, ethyl cellulose and cellulose acetate; tragacanth powder; malt; gelatin; talc; excipients such as cocoa butter and suppository wax; oils such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil and soybean oil; glycols; propylene glycol and the like; esters such as ethyl oleate and ethyl laurate; agar; buffering agents such as magnesium hydroxide and aluminum hydroxide; alginic acid; pyrogen-free water; isotonic saline; Ringer's solution; ethyl alcohol and phosphate buffer solutions, and other non-toxic compatible lubricants such as sodium lauryl sulfate and magnesium stearate, as well as coloring agents, release agents, coating agents, sweetening agents, flavoring agents and fragrances, preservatives and antioxidants may also be present in the composition at the discretion of the formulator.

[0086] Accordingly, the compounds and their physiologically acceptable salts can be formulated for administration, for example, by solid dosage, eye drops, in topical oily formulations, by injection, inhalation (either orally or nasally), implant or by oral, buccal, sublingual, parenteral or rectal administration. For techniques and formulations, generally, reference may be made to "Remington’s Pharmaceutical Sciences," (Meade Publishing Co., Easton, Pa.). The therapeutic compositions should typically be sterile and stable under the conditions of manufacture and storage.

[0087] Depending on the route of administration of the disclosed compounds and the form of the composition, the type of carrier to be used will be determined. The present composition can be in various forms suitable for, for example, systemic administration (e.g., oral, rectal, nasal, sublingual, buccal, implant or parenteral) or topical administration (e.g., transdermal, transpulmonary, nasal, otic, ophthalmic, liposomal delivery system or iontophoresis).

[0088] Carriers for systemic administration typically include at least one of diluents, lubricants, binders, disintegrants, colorants, flavorants, sweeteners, antioxidants, preservatives, lubricants, solvents, suspending agents, wetting agents, surfactants, combinations thereof, etc. All carriers are optional in the composition.

[0089] Suitable diluents include saccharides such as glucose, lactose, dextrose and sucrose; diols such as propylene glycol; calcium carbonate; sodium carbonate; sugar alcohols such as glycerin; mannitol; and sorbitol. The amount of one or more diluents in a systemic or topical composition is typically about 50% to about 90%.

[0090] Suitable lubricants include silica, talc, stearic acid and its magnesium and calcium salts, calcium sulfate; and polyethylene glycol and liquid lubricants such as vegetable oils, for example peanut oil, cottonseed oil, sesame oil, olive oil, corn oil and cocoa butter. The amount of one or more lubricants in a systemic or topical composition is typically about 5% to about 10%.

[0091] Suitable binders include polyvinylpyrrolidone; aluminum magnesium silicate; starches such as corn starch and potato starch; gelatin; tragacanth; and cellulose and its derivatives, such as sodium carboxymethyl cellulose, ethyl cellulose, methyl cellulose, microcrystalline cellulose and sodium carboxymethyl cellulose. The amount of one or more binders in a systemic composition is typically about 5% to about 50%.

[0092] Suitable disintegrants include agar, alginic acid and its sodium salts, effervescent mixtures, croscarmellose, crospovidone, sodium carboxymethyl starch, sodium starch glycolate, clays, and ion exchange resins. The amount of one or more disintegrants in a systemic or topical composition is typically from about 0.1% to about 10%.

[0093] Suitable colorants include colorants such as FD&C dyes. When used, the amount of colorant in a systemic or topical composition is typically from about 0.005% to about 0.1%.

[0094] Suitable flavoring agents include menthol, peppermint, and fruit flavoring agents. The amount of one or more flavoring agents in a systemic or topical composition, when used, is typically from about 0.1% to about 1.0%.

[0095] Suitable sweetening agents include aspartame and saccharin. The amount of one or more sweetening agents in a systemic or topical composition is typically from about 0.001% to about 1%.

[0096] Suitable antioxidants include butylated hydroxyanisole ("BHA"), butylated hydroxytoluene ("BHT"), and vitamin E. The amount of one or more antioxidants in a systemic or topical composition is typically from about 0.1% to about 5%.

[0097] Suitable preservatives include benzalkonium chloride, methylparaben, and sodium benzoate. The amount of one or more preservatives in a systemic or topical composition is typically from about 0.01% to about 5%.

[0098] Suitable lubricants include silicon dioxide. The amount of one or more lubricants in a systemic or topical composition is typically from about 1% to about 5%.

[0099] Suitable solvents include water, isotonic saline, ethyl oleate, glycerin, castor oil hydroxide, ethanol, alcohols such as polyols (such as glycerol, propylene glycol, polyethylene glycol and the like), and phosphate buffer solutions. The amount of one or more solvents in the systemic or topical composition is typically from about 0 to about 100%. In one embodiment, the compounds of the invention are formulated in 2% glycerol in sterile water. In a further embodiment, the compound UM-1007 is formulated in 2% glycerol in sterile water. The formulation of 2% glycerol in sterile water can be for parenteral administration.

[0100] Suitable suspending agents include AVICEL RC-591 (supplied by FMC Corporation, Philadelphia, PA) and sodium alginate. The amount of one or more suspending agents in the systemic or topical composition is typically from about 1 to about 8%.

[0101] Suitable surfactants include lecithin, polysorbate 80 and sodium lauryl sulfate and the TWEENS from Atlas Powder Company, Wilmington, Delaware. Suitable surfactants include those disclosed in C.T.F.A. Cosmetic Ingredient Handbook, 1992, pp. 587-592; Remington’s Pharmaceutical Sciences, 15th Ed. 1975, pp. 335-337; and McCutcheon’s Volume 1, Emulsifiers & Detergents, 1994, North American Edition, pp. 236-239. The amount of one or more surfactants in the systemic or topical composition is typically from about 0.1% to about 5%.

[0102] The amounts of the components in the systemic composition may vary depending on the type of systemic composition to be prepared. Generally, the systemic composition comprises from 0.01% to 50% of an active compound (e.g., a compound of formula (I)) and from 50% to 99.99% of one or more carriers. Compositions for parenteral administration typically comprise from 0.1% to 10% of the active ingredient and a carrier comprising from 90% to 99.9% of a diluent and a solvent.

[0103] Compositions for oral administration may have various dosage forms. For example, solid forms include tablets, capsules, granules and bulk powders. These oral dosage forms typically contain a safe and effective amount, usually at least about 5% and more particularly from about 25% to about 50% of the active ingredient. Oral dosage compositions contain from about 50% to about 95% of a carrier and more particularly from about 50% to about 75%.

[0104] Tablets can be compressed tablets, rubbed tablets, enteric-coated tablets, sugar-coated tablets, film-coated tablets or multi-layer tablets. Tablets typically contain the active ingredient and a carrier comprising components selected from diluents, lubricants, binders, disintegrants, colorants, flavorants, sweeteners, glidants and combinations thereof. Specific diluents include calcium carbonate, sodium carbonate, mannitol, lactose and cellulose. Specific binders include starch, gelatin and sucrose. Specific disintegrants include alginic acid and croscarmellose. Specific lubricants include magnesium stearate, stearic acid and talc. Specific colorants are FD&C dyes, which may be added for appearance. Chewable tablets preferably contain sweeteners such as aspartame and saccharin or flavorants such as menthol, peppermint, fruit flavorants or combinations thereof.

[0105] Capsules (including implants, timed release, and sustained release formulations) typically contain an active compound (e.g., a compound of formula (I)) and a carrier comprising one or more of the diluents disclosed above in a capsule containing gelatin. Granules typically contain the disclosed compound and a lubricant such as silicon dioxide which preferably improves flow properties. Implants can be of the biodegradable or non - biodegradable type.

[0106] The selection of components in the carrier for oral compositions depends on secondary considerations which are not critically important for the purposes of the present invention, such as taste, cost, and storage stability.

[0107] Solid compositions can be coated by conventional methods, typically with a pH - or time - dependent coating, and the disclosed compound will be released in the gastrointestinal tract in the vicinity of the desired application or at various locations and times to extend the desired action. Coatings typically include one or more components selected from the group consisting of cellulose acetate phthalate, polyvinyl acetate phthalate, hydroxypropyl methylcellulose phthalate, ethyl cellulose, EUDRAGIT® coatings (available from Evonik Industrie, Essen, Germany), waxes, and shellac.

[0108] Compositions for oral administration can have a liquid form. For example, suitable liquid forms include aqueous solutions, emulsions, suspensions, solutions reconstituted from non - effervescent granules, suspensions reconstituted from non - effervescent granules, effervescent preparations reconstituted from effervescent granules, elixirs, tinctures, syrups, etc. Liquid oral dosage compositions typically contain the disclosed compound and a carrier, i.e., a carrier selected from diluents, colorants, flavorants, sweeteners, preservatives, solvents, suspending agents, and surfactants. Oral liquid compositions preferably contain one or more components selected from colorants, flavorants, and sweeteners.

[0109] Other compositions useful for achieving systemic delivery of the subject compounds include sublingual, buccal and nasal dosage forms. Such compositions typically include a diluent, including sucrose, sorbitol and mannitol; and one or more soluble filler substances, such as binders including acacia, microcrystalline cellulose, carboxymethyl cellulose and hydroxypropyl methyl cellulose. Such compositions may further include lubricants, colorants, flavorants, sweeteners, antioxidants and lubricants.

[0110] The disclosed compounds can be administered topically. Topical compositions that can be applied topically to the skin can be in any form including solids, solutions, oils, creams, ointments, gels, lotions, shampoos, leave-in and rinse-off hair conditioners, emulsions, cleansing solutions, moisturizers, sprays, skin patches, etc. The topical composition includes the disclosed compound (e.g., a compound of formula (I)) and a carrier. The carrier of the topical composition preferably promotes the penetration of the compound into the skin. The carrier may further include one or more optional components.

[0111] The amount of carrier used in combination with the disclosed compound is sufficient to provide a composition in a practical amount for administration per unit dose of the compound. Techniques and compositions for creating dosage forms useful in the methods of the present invention are described in the following references: Modern Pharmaceutics, Chapters 9 and 10, Banker & Rhodes, eds. (1979); Lieberman et al., Pharmaceutical Dosage Forms: Tablets (1981); and Ansel, Introduction to Pharmaceutical Dosage Forms, 2nd Ed., (1976).

[0112] The carrier can include a single component or a combination of two or more components. In a topical composition, the carrier includes a topical carrier. Suitable topical carriers include one or more components selected from phosphate buffered saline, isotonic water, deionized water, monofunctional alcohols, symmetrical alcohols, aloe vera gel, allantoin, glycerin, vitamin A and E oils, mineral oil, propylene glycol, PPG-2 myristyl propionate, dimethyl isosorbide, castor oil, combinations thereof, and the like. More specifically, carriers for skin application include propylene glycol, dimethyl isosorbide, and water, and even more specifically, phosphate buffered saline, isotonic water, deionized water, monofunctional alcohols, and symmetrical alcohols.

[0113] The carrier of the topical composition can further include one or more components selected from skin softeners, propellants, solvents, humectants, thickeners, powders, fragrances, dyes, and preservatives, all of which are optional.

[0114] Suitable skin softeners include stearyl alcohol, glyceryl monolignolate, glyceryl monostearate, propane-1,2-diol, butane-1,3-diol, mink oil, cetyl alcohol, isopropyl isostearate, stearic acid, isobutyl palmitate, isocetyl stearate, oleyl alcohol, isopropyl laurate, hexyl laurate, decyl oleate, octadecan-2-ol, isocetyl alcohol, cetyl palmitate, di-n-butyl sebacate, isopropyl myristate, isopropyl palmitate, isopropyl stearate, butyl stearate, polyethylene glycol, triethylene glycol, lanolin, sesame oil, coconut oil, peanut oil, castor oil, acetylated lanolin alcohols, petroleum, mineral oil, butyl myristate, isostearic acid, palmitic acid, isopropyl linoleate, lauryl lactate, myristyl lactate, decyl oleate, myristyl myristate, and combinations thereof. Specific skin softeners for the skin include stearyl alcohol and polydimethylsiloxane. The amount of one or more skin softeners in the topical composition of the skin base is typically from about 5% to about 95%.

[0115] Suitable propellants include propane, butane, isobutane, dimethyl ether, carbon dioxide, nitrous oxide, and combinations thereof. The amount of one or more propellants in the topical composition is typically from about 0% to about 95%.

[0116] Suitable solvents include water, ethyl alcohol, methylene chloride, isopropanol, castor oil, ethylene glycol monoethyl ether, diethylene glycol monobutyl ether, diethylene glycol monoethyl ether, dimethyl sulfoxide, dimethylformamide, tetrahydrofuran, and combinations thereof. Specific solvents include ethyl alcohol and homotopic alcohols. The amount of one or more solvents in the topical composition is typically from about 0% to about 95%.

[0117] Suitable humectants include glycerin, sorbitol, sodium 2-pyrrolidone-5-carboxylate, soluble collagen, dibutyl phthalate, gelatin, and combinations thereof. A specific humectant is glycerin. The amount of one or more humectants in the topical composition is typically from 0% to 95%.

[0118] The amount of one or more thickeners in the topical composition is typically from about 0% to about 95%.

[0119] Suitable powders include β-cyclodextrins, hydroxypropyl cyclodextrins, chalk, talc, fuller's earth, kaolin, starch, gums, colloidal silicon dioxide, sodium polyacrylate, tetraalkylammonium smectites, trialkylarylammonium smectites, chemically modified aluminum magnesium silicate, organically modified montmorillonite clay, hydrated aluminum silicate, fumed silica, carboxyvinyl polymer, sodium carboxymethyl cellulose, ethylene glycol monostearate, and combinations thereof. The amount of one or more powders in the topical composition is typically from 0% to 95%.

[0120] The amount of fragrance in the topical composition is typically from about 0% to about 0.5%, particularly from about 0.001% to about 0.1%.

[0121] Suitable pH adjusting additives include HCl or NaOH in an amount sufficient to adjust the pH of the topical pharmaceutical composition.

[0122] b. Adjuvants and Vaccine Compositions The compound can be incorporated into adjuvant compositions and vaccine compositions. The adjuvant composition can induce an immune response. In some embodiments, the adjuvant composition induces a Th1-type immune response.

[0123] The vaccine composition may further contain an antigen. Suitable antigens include microbial pathogens, bacteria, viruses, proteins, glycoproteins, lipoproteins, peptides, glycopeptides, lipopeptides, toxoids, carbohydrates, and tumor-specific antigens. A mixture of two or more antigens may be used. In some embodiments, the antigen is derived from bacteria, viruses, bacteriophages, fungi, prions, neoplasms, autoantigens, animals, plants, recombinants, or synthetic substances. The antigens include the bacteria, viruses, fungi, plants, and cancer / tumor antigens described in WO 2019 / 157509 pamphlet, which is hereby incorporated by reference herein.

[0124] The adjuvant and the vaccine composition may contain a "effective amount" of the disclosed compound. In the context of an adjuvant or a vaccine composition, an "effective amount" refers to an amount effective to achieve the desired result (e.g., enhancing the immune response to one or more antigens) at the required dosage and duration. The immune response can be measured, for example, by measuring the antibody titer against the antigen, evaluating the ability of the vaccine containing the compound to immunize the host in response to a disease or antigen challenge, etc. For example, when an "effective amount" of a compound or composition is administered to a subject, one or more antibody titers increase by 10% or more, 20% or more, 30% or more, 40% or more, 50% or more, 50% or more, 70% or more, 80% or more, 90% or more, or 100% or more compared to a non-immunized control.

[0125] Vaccine preparation is a well-developed technology, and general guidelines for the preparation and formulation of vaccines are readily available from any of a variety of information sources. One such example is New Trends and Developments in Vaccines, edited by Voller et al., University Park Press, Baltimore, Md., U.S.A. 1978.

[0126] The vaccine compositions of the present disclosure may also contain other compounds which may be biologically active or inactive. For example, one or more immunogenic portions of other tumor antigens may be incorporated into the fusion polypeptide or may be present within the vaccine composition as separate compounds. The polypeptide, although not essential, may be conjugated to other macromolecules as described, for example, in U.S. Pat. Nos. 4,372,945 and 4,474,757. The vaccine compositions may generally be used for prophylactic and therapeutic purposes.

[0127] In one embodiment, the antigen in the vaccine composition is in the form of a peptide, polypeptide, protein or an immunogenic portion thereof. An "immunogenic portion" as used herein is a portion of a protein that is recognized (i.e., specifically binds) by B cell and / or T cell surface antigen receptors. Such an immunogenic portion generally comprises at least 5 amino acid residues, more preferably at least 10 and even more preferably at least 20 amino acid residues of the antigen protein or a variant thereof.

[0128] The immunogenic portion of an antigen polypeptide can generally be identified using well-known techniques, such as those summarized in Paul, Fundamental Immunology, 3rd ed., 243-247 (Raven Press, 1993) and the references cited therein. Such techniques include screening the polypeptide for its ability to react with antigen-specific antibodies, antisera, and / or T cell lines or clones. As used herein, antisera and antibodies are "antigen-specific" if they specifically bind to the antigen (i.e., they react with that protein in an ELISA or other immunoassay and do not show a detectable reaction with unrelated proteins). Such antisera and antibodies can be prepared as described herein and using well-known techniques. The immunogenic portion of a protein is a portion that reacts with such antisera and / or T cells at a level that does not substantially fall below the reactivity of the full-length polypeptide (e.g., in an ELISA and / or T cell reactivity assay). Such an immunogenic portion can react at a level equal to or higher than the reactivity of the full-length polypeptide in such an assay. Such screening can generally be carried out using methods well known to those skilled in the art, such as those described in Harlow and Lane, Antibodies: A Laboratory Manual, Cold Spring Harbor Laboratory, 1988. For example, by immobilizing the polypeptide on a solid support and contacting it with patient serum, the antibodies in the serum can be bound to the immobilized polypeptide. Next, the unbound serum is removed, and the bound antibodies can be detected, for example 125 using I-labeled protein A.

[0129] In some embodiments, the antigen is a hapten, a hapten conjugated to a carrier protein, a polypeptide or other polymer or a derivative thereof. Haptens mainly include small molecules that can elicit an immune response only when bound to a large carrier such as a protein. Haptens can be linked to carrier proteins and additional moieties. Exemplary haptens are aniline, o-, m- and p-aminobenzoic acid, quinone, histamine-succinyl-glycine (HSG), hydralazine, halogen, indium-DTPA, fluorescein, biotin, digoxigenin, theophylline and dinitrophenol.

[0130] Common carrier proteins include serum globulin, albumin, ovalbumin and many others as well as synthetic polypeptides such as poly L-glutamic acid. Polysaccharides and liposomes can also be used.

[0131] In some embodiments, the antigen is an allergen. Allergens include naturally occurring proteins and other small molecules that have been reported to induce an allergy, i.e., an IgE-mediated reaction, to repeated exposure to them in an individual. Examples of naturally occurring allergens include pollen allergens (tree, weed, herb and grass pollen allergens), mite allergens (e.g., from house dust mites and storage mites), insect allergens (allergens caused by inhaled antigens, saliva and venom), animal allergens such as those derived from, for example, dogs, cats, horses, rats, mice, etc., such as those derived from saliva, hair and dander, fungal allergens as well as food allergens. Allergens also include drugs and latex.

[0132] In another embodiment, the compounds or adjuvant compositions described herein can be used in the preparation of DNA-based vaccine compositions. Exemplary vaccines of this type contain DNA encoding one or more polypeptide antigens such that the antigen is produced in situ. The DNA can be present in any of a variety of delivery systems known to those of skill in the art, including nucleic acid expression systems, bacterial and viral expression systems. Many gene delivery techniques are well known in the art, such as those described by Rolland, Crit. Rev. Therap. Drug Carrier Systems 15:143-198, 1998 and the references cited therein. Suitable nucleic acid expression systems contain the DNA sequences necessary for expression in the patient (such as a suitable promoter and termination signal). Bacterial delivery systems involve the administration of bacteria (such as Bacillus-Calmette-Guerin) that express the immunogenic portion of the polypeptide or secrete such epitopes on their cell surface. In a preferred embodiment, the DNA is introduced using a viral expression system (e.g., vaccinia or other poxviruses, retroviruses or adenoviruses), which typically involves the use of non-pathogenic (defective) replication-competent viruses.For exemplary systems, see, for example, Fisher-Hoch et al., Proc. Natl. Acad. Sci. USA 86:317-321, 1989; Flexner et al., Ann. N.Y. Acad. Sci. 569:86-103, 1989; Flexner et al., Vaccine 8:17-21, 1990; U.S. Patent Nos. 4,603,112, 4,769,330 and 5,017,487; International Publication No. WO 89 / 01973; U.S. Patent No. 4,777,127; British Patent No. 2,200,651; European Patent No. 0,345,242; International Publication No. WO 91 / 02805; Berkner, Biotechniques 6:616-627, 1988; Rosenfeld et al., Science 252:431-434, 1991; Kolls et al., Proc. Natl. Acad. Sci. USA 91:215-219, 1994; Kass-Eisler et al., Proc. Natl. Acad. Sci. USA 90:11498-11502, 1993; Guzman et al., Circulation 88:2838-2848, 1993; and Guzman et al., Cir. Res. 73:1202-1207, 1993. Techniques for introducing DNA into such expression systems are well known to those of skill in the art.

[0133] Alternatively, the DNA can be "naked", as described, for example, in Ulmer et al., Science 259:1745-1749, 1993 and reviewed by Cohen, Science 259:1691-1692, 1993. Uptake of naked DNA can be increased by coating the DNA onto biodegradable beads that are efficiently transported into cells.

[0134] Furthermore, it will be apparent that the vaccine may contain a pharmaceutically acceptable salt of the desired antigen. For example, such salts can be prepared from pharmaceutically acceptable non-toxic bases including organic bases (e.g., salts of primary, secondary and tertiary amines and basic amino acids) and inorganic bases (e.g., sodium, potassium, lithium, ammonium, calcium and magnesium salts).

[0135] The adjuvant system can exhibit a strong adjuvant effect when administered over a wide range of dosages and ratios.

[0136] The amount of antigen in each vaccine dose is generally selected as an amount that induces an immune defense response without significant adverse side effects typical of a vaccine. Such amount will vary depending specifically on which immunogen is used and what form it is in. Of course, the dosage administered can depend on age, body weight, type of concomitant therapy if any, and the nature of the antigen being administered.

[0137] The immunogenic activity of a given amount of vaccine composition can be readily determined, for example, by monitoring an increase in the titer of antibodies to the antigen used in the vaccine composition (Dalsgaard, K. Acta Veterinia Scandinavica 69:1-40 (1978)). Another common method involves intradermally injecting various amounts of the vaccine composition into CD-1 mice, later collecting serum from the mice and testing for anti-immunogen antibodies, for example, by ELISA. These and other similar techniques will be apparent to those skilled in the art.

[0138] An antigen can be derived from and / or isolated from essentially any desired source depending on the infectious disease, autoimmune disease, condition, cancer, pathogen or disease to be treated with a given vaccine composition. By way of example, an antigen can be derived from a viral source such as influenza virus, feline leukemia virus, feline immunodeficiency virus, human HIV-1, HIV-2, herpes simplex virus type 2, human cytomegalovirus, hepatitis A, B, C or E, respiratory syncytial virus, human papillomavirus, rabies, measles or foot-and-mouth disease virus. Exemplary antigens can be derived from a bacterial source such as anthrax, diphtheria, Lyme disease, malaria, tuberculosis, leishmaniasis, Trypanosoma cruzi, Ehrlichia, Candida, etc., or can also be derived from protozoa such as Babesia bovis or Plasmodium. One or more antigens will typically comprise natural or synthetic amino acids, for example in the form of a peptide, polypeptide or protein, may include polysaccharides, or may be a mixture thereof. Exemplary antigens can be isolated from natural sources, synthesized using solid-phase synthesis, or obtained by recombinant DNA techniques.

[0139] In another embodiment, tumor antigens can be used in a vaccine composition for the prevention and / or treatment of cancer. Tumor antigens are surface molecules that are differentially expressed in tumor cells compared to non-tumor tissues. Tumor antigens are those that immunologically distinguish tumor cells from normal cells and provide diagnostic and therapeutic targets for human cancers. Tumor antigens are characterized either as membrane proteins or as altered carbohydrate molecules of glycoproteins or glycolipids on the cell surface. Cancer cells often have truncated epidermal growth factor, folate-binding protein, epithelial mucin, melanoferrin, carcinoembryonic antigen, prostate-specific membrane antigen, HER 2-neu, etc. (these are candidates for use in therapeutic cancer vaccines) have specific tumor antigens on their surface. Since the tumor antigens are normal or related to normal components of the body, the immune system often cannot produce an effective immune response against such antigens to destroy tumor cells. To achieve such a response, the adjuvant systems described herein can be utilized. As a result, the exogenous protein can enter the processing pathway of the endogenous antigen, leading to the production of cytolytic or cytotoxic T cells (CTLs). This adjuvant effect promotes the production of antigen-specific CTLs, which search for and destroy tumor cells that carry one or more of the tumor antigens used for immunization on their surface. Exemplary cancer types for which this approach can be used include prostate cancer, colon cancer, breast cancer, ovarian cancer, pancreatic cancer, brain cancer, head and neck cancer, melanoma, leukemia, lymphoma, and the like.

[0140] In one embodiment, the antigen present in the vaccine composition is not a foreign antigen but a self-antigen, i.e., this vaccine composition is directed against autoimmune diseases. Examples of autoimmune diseases include type 1 diabetes, common organ-specific autoimmunity, neurological diseases, rheumatic diseases / connective tissue diseases, autoimmune hemocytopenia and related autoimmune diseases. Such common organ-specific autoimmunity may include thyroiditis (Graves' disease + Hashimoto's disease), gastritis, adrenalitis (Addison's disease), oophoritis, primary biliary cirrhosis, myasthenia gravis, gonadal insufficiency, hypoparathyroidism, alopecia, malabsorption syndrome, pernicious anemia, hepatitis, anti-receptor antibody diseases and vitiligo. Such neurological diseases may include schizophrenia, Alzheimer's disease, depression, hypopituitarism, diabetes insipidus, Sjogren's syndrome and multiple sclerosis. Such rheumatic diseases / connective tissue diseases may include rheumatoid arthritis, systemic lupus erythematosus (SLE) or lupus, scleroderma, polymyositis, inflammatory bowel disease, dermatomyositis, ulcerative colitis, Crohn's disease, vasculitis, psoriatic arthritis, exfoliative psoriatic dermatitis, pemphigus vulgaris, Sjogren's syndrome. Other autoimmune-related diseases may include autoimmune retinal uveitis, glomerulonephritis, post-myocardial infarction syndrome, pulmonary hemosiderosis, amyloidosis, sarcoidosis, aphthous stomatitis and other immune-related diseases as presented herein and known in the relevant technical fields.

[0141] In one embodiment, an antigen can be covalently bound to an adjuvant such as a compound of formula I to create a separate molecule that can exhibit enhanced adjuvant effects on the antigen, and this adjuvant effect can be higher than the adjuvant effect achievable in the absence of such a covalent bond, such as in a mixture of components (i.e., a mixture of the antigen and the compound of formula (I)). The covalent bond can be achieved by a reaction via a functional group; for example, in the case of a compound of formula I, by a reaction via a carboxylic acid group, a hydroxyl group, or an aldehyde functional group. For such covalently bound antigens, further enhancement of the adjuvant effect can be achieved by incorporating an inorganic salt adjuvant together with such a compound. The inorganic salt adjuvant preferably includes aluminum hydroxide or aluminum phosphate, but other known inorganic salt adjuvants such as calcium phosphate, zinc hydroxide, or calcium hydroxide can be used.

[0142] The vaccine composition can be formulated for any suitable route of administration, including, for example, topical, oral, nasal, intravenous, vaginal, cutaneous, sublingual, intracranial, intradermal, intraperitoneal, subcutaneous, intramuscular administration, or inhalation, and can be administered as such. For parenteral administration such as subcutaneous injection, the carrier preferably includes water, physiological saline, alcohol, fat, wax, or a buffer solution. For oral administration, any of the above carriers or a solid carrier such as mannitol, lactose, starch, magnesium stearate, sodium saccharin, talc, cellulose, glucose, sucrose, and magnesium carbonate can be used.

[0143] In an exemplary embodiment, the vaccine formulation is administered to mucosa, particularly the oral cavity and preferably to sublingual sites, to elicit an immune response. In many cases, oral administration may be preferred over traditional parenteral delivery due to the ease and convenience offered by non-invasive administration techniques. Moreover, this approach further provides a means to elicit mucosal immunity that may often be difficult to achieve with traditional parenteral delivery and may confer protection from airborne pathogens and / or allergens. An additional advantage of oral administration is that sublingual vaccine delivery may improve patient compliance, especially for pediatric applications or for applications that traditionally require multiple injections over a long period of time, such as with allergy desensitization therapy.

[0144] The vaccine composition may also include buffers (e.g., neutral buffered saline or phosphate buffered saline), carbohydrates (e.g., glucose, mannose, sucrose or dextran), mannitol, proteins, polypeptides or amino acids such as glycine, antioxidants, bacteriostatic agents, chelating agents such as EDTA or glutathione, adjuvants (e.g., aluminum hydroxide), solutes that render the formulation isotonic, hypotonic or slightly hypotonic with the blood of the recipient, suspending agents, thickening agents and / or preservatives. Alternatively, the vaccine composition may be formulated as a lyophilizate. The compounds may also be encapsulated in liposomes using well-known techniques.

[0145] The vaccine composition may also include other adjuvants or immunoeffectors. Suitable adjuvants include, for example, Freund's incomplete and complete adjuvants (Difco Laboratories, Detroit, Mich.); Merck Adjuvant 65 (Merck and Company, Inc., Rahway, NJ); AS-2 (SmithKline Beecham); inorganic salts (e.g., aluminum, silica, kaolin, and carbon); aluminum hydroxide gel (alum), AlK(SO 4 ) 2 , AlNa(SO 4 ) 2 , AlNH 4 (SO 4 ) and Al(OH)3 aluminum salts such as; calcium salts (e.g., Ca 3 (PO 4 ) 2) iron or zinc; an insoluble suspension of acylated tyrosine; acylated saccharides; cation-derivatized or anion-derivatized polysaccharides; polynucleotides (e.g., poly IC and poly AU acid); polyphosphazenes; cyanoacrylates; poly(lactic acid-co-glycoside); biodegradable microspheres; liposomes; lipid A and its derivatives; monophosphoryl lipid A; wax D derived from Mycobacterium tuberculosis and substances found in members of Corynebacterium parvum, Bordetella pertussis, and Brucella; bovine serum albumin; diphtheria toxoid; tetanus toxoid; edestin; keyhole limpet hemocyanin; Pseudomonas aeruginosa toxin A; cholera genoid; cholera toxin; pertussis toxin; viral proteins; and are commercially available as Quil A. Aminoalkyl glucosamine phosphate compounds can also be used (see, for example, WO 98 / 50399, U.S. Patent No. 6,113,918 (granted from U.S. Patent Application No. 08 / 853,826), and U.S. Patent Application No. 09 / 074,720). In addition, adjuvants such as cytokines (e.g., GM-CSF or interleukin-2, -7, or -12), interferons, or tumor necrosis factors can also be used as adjuvants. Protein and polypeptide adjuvants can be obtained from natural or recombinant sources by methods well known to those skilled in the art. When obtained from recombinant sources, the adjuvant can include a protein fragment containing at least the immunostimulatory portion of the molecule. Other known immunostimulatory macromolecules that can be used include, but are not limited to, polysaccharides, tRNA, non-metabolic synthetic polymers such as polyvinylamine, polymethacrylic acid, polyvinylpyrrolidone, a mixed polycondensate of 4',4-diaminodiphenylmethane-3,3'-dicarboxylic acid and 4-nitro-2-aminobenzoic acid (having a relatively high molecular weight) (see Sela, M., Science 166:1365-1374 (1969)) or glycolipids, lipids, or carbohydrates.

[0146] In the vaccines provided herein, the adjuvant composition is preferably designed to predominantly elicit a Th1-type immune response. High levels of Th1-type cytokines (e.g., IFN-γ, IL-2, and IL-12) tend to be favorable for the induction of a cell-mediated immune response to the administered antigen. In contrast, high levels of Th2-type cytokines (e.g., IL-4, IL-5, IL-6, IL-10, and TNF-β) tend to be favorable for the induction of a humoral immune response. After application of the vaccines as provided herein, the patient supports an immune response that includes both Th1 and Th2-type responses. Within the preferred embodiments where the response is predominantly Th1-type, the levels of Th1-type cytokines can increase to be higher than the levels of Th2-type cytokines. The levels of these cytokines can be readily evaluated using standard assays. For a review of the cytokine family, see Mosmann and Coffman, 1989, Ann. Rev. Immunol. 7:145-173.

[0147] The compositions described herein can be administered as part of a sustained release formulation (i.e., a formulation such as a capsule, sponge or gel (e.g., made of a polysaccharide) that results in a slow release of the compound after administration). Such formulations are generally prepared using well-known techniques (see, e.g., Coombes et al., Vaccine 14:1429-1438, 1996) and can be administered, for example, orally, rectally or by subcutaneous implantation or implantation at the desired target site. The sustained release formulation can contain a polypeptide, polynucleotide or antibody dispersed in a carrier matrix and / or placed within a reservoir surrounded by a rate-limiting membrane. The carrier used in such a formulation can be biocompatible and biodegradable; preferably, the formulation provides a relatively constant level of release of the active ingredient. Such carriers include microparticles such as poly(lactide-co-glycolide), polyacrylate, latex, starch, cellulose, dextran and the like. Other delayed release carriers include supramolecular biovectors, which include a non-liquid hydrophilic core (e.g., a cross-linked polysaccharide or oligosaccharide) and, optionally, an outer layer containing an amphiphilic compound such as a phospholipid (see, e.g., U.S. Patent No. 5,151,254 and International Publications Nos. 94 / 20078, 94 / 23701 and 96 / 06638). The amount of the active compound contained in the sustained release formulation will vary depending on the site of implantation, the rate of release and the expected duration, as well as the nature of the condition to be treated or prevented.

[0148] In pharmaceutical compositions and vaccines, any of a variety of known delivery vehicles can be used to promote the generation of an antigen-specific immune response that targets cells. Examples of delivery vehicles include antigen-presenting cells (APCs), such as dendritic cells, macrophages, B cells, monocytes, and other cells that can be engineered to be efficient APCs. Such cells are not essential, but can be genetically modified to have increased antigen-presenting ability, improved activation and / or maintenance of T cell responses, have an anti-target effect per se, and / or be immunologically compatible with the recipient (i.e., have a matching HLA haplotype). APCs can generally be isolated from any of a variety of body fluids and organs, including tumors and peritumoral tissues, and can be autologous cells, allogeneic cells, syngeneic cells, or xenogeneic cells.

[0149] The composition can include liposomal vesicles containing a compound of formula I. Liposomes are generally made of phospholipids or other lipid substances. The preparation procedures of liposomes are well known to those skilled in the art. Any lipid capable of forming vesicles containing the compound of formula I can be used. For clinical applications, it is desirable that the lipid is non-toxic, physiologically acceptable, and metabolizable. Commonly used bilayer-forming lipids with clinical potential are phospholipids, fatty acids, sphingolipids, glycosphingolipids, and steroids. Glycerol-containing phospholipids are the most widely used components of clinically useful liposomal formulations. One commonly used example is phosphatidylcholine or lecithin. Steroids cholesterol and its derivatives are often included as components of the liposomal membrane. The tendency of liposomes to aggregate and fuse can be controlled by including a small amount of acidic or basic lipids in the formulation. The properties of liposomes containing phospholipids are determined by the chemistry of the phospholipids. Important factors to consider are the hydrocarbon chain length, the degree of unsaturation of the hydrocarbon chain, the degree of branching of the hydrocarbon chain, and the temperature of the system.

[0150] Multilamellar liposomes can be prepared by depositing a mixture of lipids as a thin film by evaporation under reduced pressure and subsequently dispersing it in an excess amount of aqueous buffer with or without an antigen. Another method is to mix an aqueous phase containing an antigen with small unilamellar liposomes and then lyophilize. When the lyophilized product is usually rehydrated with a small amount of distilled water, multilamellar liposomes are formed. The small unilamellar liposomes used in this method are prepared by dispersing lipids in an aqueous medium followed by mechanical dispersion means such as sonication, use of a high-pressure device, or solvent injection method. Large and intermediate-sized unilamellar liposomes can also be prepared by conventional techniques including detergent dialysis, extrusion under high pressure through a membrane with a small pore size, freeze-thaw followed by slow swelling, dehydration followed by rehydration and dilution, or dialysis of lipids in the presence of chaotropic ions. The size of the liposomes can be made more uniform by fractionation procedures such as centrifugation or size exclusion chromatography, homogenization, or capillary pore membrane extrusion.

[0151] 4. Method of Use The disclosed compounds and compositions can be used in a variety of methods including methods of modulating an immune response in a subject, methods of inducing or enhancing the immunogenicity of an antigen in a subject, and methods of treating, preventing, or reducing susceptibility to allergy, autoimmune disease, cancer, or bacterial, viral, or prion infections in a subject.

[0152] a. Modulation of Immune Response The disclosed compounds and compositions can be used in a method of modulating an immune response in a subject, the method comprising administering to the subject an effective amount of a compound described herein, an adjuvant composition described herein, a vaccine composition described herein, or a pharmaceutical composition described herein.

[0153] In some embodiments, the immune response in a subject is increased. In some embodiments, the subject has cancer, an autoimmune disorder, an allergy, or an infectious disease. The infectious disease can be caused by a virus, bacterium, or prion or prion-like protein.

[0154] The disclosed compounds and compositions can be used in methods of inducing an enhanced immune response in a subject.

[0155] Enhancement of the immune response can be induced by co-administering the compound or composition with an antigen. Suitable antigens include microbial pathogens, bacteria, viruses, proteins, glycoproteins, lipoproteins, peptides, glycopeptides, lipopeptides, toxoids, carbohydrates, and tumor-specific antigens. Mixtures of two or more antigens can be used.

[0156] b. Induction or enhancement of antigen immunogenicity The disclosed compounds and compositions can be used in methods of inducing or enhancing the immunogenicity of an antigen in a subject, the method comprising administering to the subject a vaccine composition comprising the antigen and an adjuvant composition comprising an effective amount of a compound or composition described herein.

[0157] Suitable antigens include microbial pathogens, bacteria, viruses, proteins, glycoproteins, lipoproteins, peptides, glycopeptides, lipopeptides, toxoids, carbohydrates, and tumor-specific antigens. Mixtures of two or more antigens can be used. In some embodiments, the antigen is derived from a bacterium, virus, bacteriophage, fungus, prion, neoplasm, autoantigen, animal, plant, recombinant, or synthetic substance.

[0158] c. Methods of treating, preventing, or reducing susceptibility to diseases and disorders The disclosed compounds and compositions can be used in a method of treating, preventing, or reducing susceptibility to a disease or disorder, the method comprising administering to a subject in need thereof a therapeutically effective amount of a compound or composition described herein.

[0159] i. Allergy In some embodiments, the disease or disorder is an allergy or an allergic disease / condition. An allergy refers to an acquired hypersensitivity to a substance (allergen). Allergic conditions include, for example, eczema, allergic rhinitis or hay fever, allergic asthma, hives (urticaria), and food allergies, as well as other atopic conditions.

[0160] The method can reduce, inhibit, or stop / prevent an allergic or inflammatory reaction (e.g., by reducing or arresting the antibody production or amount of antibodies against a specific antigen).

[0161] ii. Autoimmune disease In some embodiments, the disease or disorder is an autoimmune disease. Autoimmune diseases occur when an individual's immune system attacks its own organs or tissues, resulting in a clinical condition associated with the destruction of that tissue. Autoimmune diseases include, for example, rheumatoid arthritis, insulin-dependent diabetes, acquired immunodeficiency syndrome ("AIDS"), hemolytic anemia, rheumatic fever, Crohn's disease, Guillain-Barré syndrome, psoriasis, thyroiditis, Graves' disease, myasthenia gravis, glomerulonephritis, autoimmune hepatitis, multiple sclerosis, and systemic lupus erythematosus. The method can reduce, inhibit, or stop an autoimmune reaction.

[0162] iii. Infectious disease or infection In some embodiments, the disease or disorder is an infectious disease or infection. Infectious diseases are caused by infectious agents including, but not limited to, viruses, bacteria, fungi, protozoa, parasites, and prions or prion-like proteins. In some embodiments, the infectious disease or infection can be caused by bacteria, viruses, or prions or prion-like proteins.

[0163] Viral diseases that can be treated by the method of the present invention include, but are not limited to, those caused by hepatitis A, hepatitis B, hepatitis C, influenza, varicella, adenovirus, herpes simplex type I (HSV-I), herpes simplex type II (HSV-II), dengue fever, Ebola, Zika, rinderpest, rhinovirus, echovirus, rotavirus, respiratory syncytial virus, papillomavirus, papovavirus, cytomegalovirus, echino virus, arbovirus, huntavirus, coxsachie virus, mumps virus, measles virus, rubella virus, poliovirus, human immunodeficiency virus type I (HIV-I), and human immunodeficiency virus type II (HIV-II).

[0164] Bacterial diseases that can be treated by the method of the present invention include, but are not limited to, mycobacteria rickettsia, mycoplasma, Neisseria, cholera, gonorrhea, Lyme disease, whooping cough, plague, syphilis, tuberculosis, Rocky Mountain spotted fever, and Legionella.

[0165] iv. Cancer In some embodiments, the disease or disorder is cancer. The method can reduce or inhibit the growth of cancer cells. The method can be used in any cancer cell or in a subject having any type of cancer, such as those described by the National Cancer Institute. Exemplary cancers can include the following.

[0166] Digestive tract cancer / gastrointestinal cancer, such as anal cancer; bile duct cancer; extrahepatic bile duct cancer; appendiceal cancer; carcinoid tumor, gastrointestinal cancer; colon cancer; colorectal cancer including pediatric colorectal cancer; esophageal cancer including pediatric esophageal cancer; gallbladder cancer; gastric cancer (gastric cancer (stomach cancer)) including pediatric gastric cancer (gastric cancer) (stomach cancer); hepatocellular carcinoma (liver cancer) including adult (primary) hepatocellular carcinoma (liver cancer) and pediatric (primary) hepatocellular carcinoma (liver cancer); pancreatic cancer including pediatric pancreatic cancer; sarcoma, rhabdomyosarcoma; pancreatic islet cell carcinoma; rectal cancer; and small intestine cancer, etc.

[0167] Endocrine cancer, such as pancreatic islet cell carcinoma (pancreatic endocrine part); adrenocortical carcinoma including pediatric adrenocortical carcinoma; gastrointestinal carcinoid tumor; parathyroid cancer; pheochromocytoma; pituitary tumor; thyroid cancer including pediatric thyroid cancer; pediatric multiple endocrine neoplasia syndrome; and pediatric carcinoid tumor, etc.

[0168] Eye cancer, such as intraocular melanoma; and retinoblastoma, etc.

[0169] Musculoskeletal cancer, such as Ewing family tumor; osteosarcoma / malignant fibrous histiocytoma of bone; pediatric rhabdomyosarcoma; soft tissue sarcoma including adult and pediatric soft tissue sarcoma; clear cell sarcoma of tendon sheath; and uterine sarcoma, etc.

[0170] Breast cancer, such as breast cancer including pediatric and male breast cancer and breast cancer during pregnancy, etc.

[0171] Neural cancer, such as pediatric brainstem glioma; brain tumor; pediatric cerebellar astrocytoma; pediatric cerebral astrocytoma / malignant glioma; pediatric ependymoma; pediatric medulloblastoma; pediatric pineal and supratentorial undifferentiated neuroectodermal tumor; pediatric visual pathway and hypothalamic glioma; other pediatric brain cancers; adrenocortical carcinoma; central nervous system lymphoma, primary; pediatric cerebellar astrocytoma; neuroblastoma; craniopharyngioma; spinal cord tumor; central nervous system atypical teratoid / rhabdoid tumor; central nervous system fetal tumor; and pediatric supratentorial undifferentiated neuroectodermal tumor and pituitary tumor, etc.

[0172] Urogenital cancers, such as bladder cancer including pediatric bladder cancer; renal cell carcinoma (kidney cancer); ovarian cancer including pediatric ovarian cancer; ovarian epithelial cancer; low malignant potential ovarian tumors; penile cancer; prostate cancer; renal cell carcinoma including pediatric renal cell carcinoma; transitional cell carcinoma of the renal pelvis and ureter; testicular cancer; urethral cancer; vaginal cancer; vulvar cancer; cervical cancer; Wilms tumor and other pediatric kidney tumors; endometrial cancer; and gestational trophoblastic tumors, etc.; germ cell cancers, such as pediatric extracranial germ cell tumors; extragonadal germ cell tumors; ovarian germ cell tumors, etc.

[0173] Head and neck cancers, such as lip and oral cavity cancer; oral cavity cancer including pediatric oral cavity cancer; hypopharyngeal cancer; laryngeal cancer including pediatric laryngeal cancer; metastatic squamous cell carcinoma of the neck of unknown primary origin; oral cavity cancer; nasal cavity and paranasal sinus cancer; nasopharyngeal cancer including pediatric nasopharyngeal cancer; oropharyngeal cancer; parathyroid cancer; pharyngeal cancer; salivary gland cancer including pediatric salivary gland cancer; laryngopharyngeal cancer; and thyroid cancer, etc.

[0174] Blood cancers / blood cell cancers, such as leukemia (e.g., acute lymphoblastic leukemia including adult and pediatric acute lymphoblastic leukemia; acute myeloid leukemia including adult and pediatric acute myeloid leukemia; chronic lymphocytic leukemia; chronic myelogenous leukemia; and hairy cell leukemia); lymphoma (e.g., AIDS-related lymphoma; cutaneous T-cell lymphoma; Hodgkin lymphoma including adult and pediatric Hodgkin lymphoma and Hodgkin lymphoma during pregnancy; non-Hodgkin lymphoma including adult and pediatric non-Hodgkin lymphoma and non-Hodgkin lymphoma during pregnancy; mycosis fungoides; Sézary syndrome; Waldenström macroglobulinemia; and primary central nervous system lymphoma); and other blood cancers (e.g., chronic myeloproliferative disorders; multiple myeloma / plasma cell tumors; myelodysplastic syndromes; and myelodysplastic / myeloproliferative diseases), etc.

[0175] Lung cancer, such as non-small cell lung cancer; and small cell lung cancer, etc.

[0176] Respiratory cancers, such as adult malignant mesothelioma; pediatric malignant mesothelioma; malignant thymoma; pediatric thymoma; thymic carcinoma; bronchial adenoma / carcinoid including pediatric bronchial adenoma / carcinoid; pleuropulmonary blastoma; non-small cell lung cancer; and small cell lung cancer, etc.

[0177] Skin cancer, such as Kaposi's sarcoma; Merkel cell carcinoma; melanoma; and pediatric skin cancer, etc.

[0178] AIDS-related malignancies.

[0179] Other pediatric cancers, rare pediatric cancers, and cancers of unknown primary site.

[0180] Metastases of the aforementioned cancers.

[0181] 5. Kit In one aspect, the present disclosure provides a kit comprising a composition comprising at least one disclosed compound or a pharmaceutically acceptable salt thereof, or a compound or a pharmaceutically acceptable salt thereof, and (a) at least one antigen; (b) at least one additional therapeutic agent; and (c) instructions regarding the administration of the compound or composition and one or more of the above.

[0182] In some embodiments, at least one disclosed compound and at least one antigen or at least one additional therapeutic agent are co-formulated. In some embodiments, at least one disclosed compound and at least one antigen or at least one additional therapeutic agent are co-packaged. The kit may also include compounds and / or products that are packaged together with, co-formulated with, and / or co-delivered with other components. For example, a drug manufacturer, drug seller, physician, pharmacy, or pharmacist can provide a kit comprising a disclosed compound and / or product and another component for delivery to a patient.

[0183] The disclosed kit can be used in connection with the disclosed methods of use.

[0184] The kit may further include information, instructions, or both, that the use of the kit will result in an increase in immunity against a specific pathogen in mammals (particularly humans). The information and instructions can be in the form of words, pictures, or both. Additionally or alternatively, the kit may include a compound, a composition, or both; and preferably information, instructions, or both regarding the method of administration of the compound or composition, along with the benefit of treating or preventing a medical condition in mammals (e.g., humans).

[0185] The compounds and methods of the present disclosure can be better understood by reference to the following examples, which are intended to be illustrative of the scope of the present disclosure and not limiting thereof.

Examples

[0186] Other suitable modifications and variations of the present disclosure described herein are readily applicable and easily determinable, and it will be immediately apparent to those skilled in the art that they can be made using suitable equivalents without departing from the scope of the present disclosure or the aspects and embodiments disclosed herein. Although the present disclosure has been described in detail, it will be more clearly understood by reference to the following examples, which are only intended to illustrate some aspects and embodiments of the present disclosure and should not be regarded as limiting the scope of the present disclosure. The disclosures of all academic journals, U.S. patents, and publications referred to herein are incorporated by reference in their entirety.

[0187] The abbreviations used in the following schemes and descriptions include the following: DCC is N,N'-dicyclohexylcarbodiimide; DMAP is N,N-dimethylpyridin-4-amine; DMF is dimethylformamide; eq is equivalent; IFNα is interferon alpha; IgG is immunoglobulin G; IL-12p70 is interleukin 12p70; Im-OTf is imidazolium trifluoromethanesulfonate; Pal is C(O)(CH 2 ) 14 CH3 wherein; PBMC is peripheral blood mononuclear cells; PEG is polyethylene glycol; PMB is para-methoxybenzyl; rt is room temperature; TBAI is tetrabutylammonium iodide; TEA is trimethylamine; THF is tetrahydrofuran; TLR is toll-like receptor; TNFα is tumor necrosis factor α; 14Dp1 is 14 days after the first immunization; 14Dp2 is 14 days after the second immunization.

[0188] Example 1. Exemplary Synthesis of Compound UM-1007 A. Synthesis of Pegylated Glycerol The required dipalmitoyl pegylated glycerol 5 was prepared in 5 steps from commercially available triethylene glycol monochloride (1).

Chemical formula

[0189] p-Methoxybenzyl chloride (3 eq) and tetra-n-butylammonium iodide (0.02 eq) were added to a solution of 1 in anhydrous DMF (2.0 M), and the solution was cooled to 0 °C. Sodium hydride (1.25 eq) was slowly added to the cold solution. After stirring at 0 °C for 30 minutes, the reaction mixture was warmed to rt. After 4 hours, the reaction mixture was slowly quenched with saturated sodium bicarbonate solution and extracted with chloroform. The organic layer was dried over sodium sulfate, filtered, and concentrated under vacuum. The crude product was purified by silica gel chromatography (10 - 40% ethyl acetate in heptane) to give 2 as a colorless oil in 90% yield. Sodium hydride (1.5 eq) was slowly added to a solution of 2 (1.5 eq), 1,2-sn-isopropylidene glycerol (1.0 eq), and tetra-n-butylammonium iodide (0.05 eq) in anhydrous THF (0.69 M). After 15 hours at 75 °C, additional 2 (0.5 eq) and sodium hydride (1.5 eq) were added and the reaction mixture was stirred at 75 °C. After 6 hours, the reaction mixture was cooled to rt, quenched by the addition of brine, and extracted 3 times with ethyl acetate. The combined organic layers were dried over sodium sulfate, filtered, and concentrated under vacuum. The crude product was purified by silica gel chromatography (20 - 80% ethyl acetate in heptane) to give 3 as a colorless oil in 74% yield. 0.4 N HCl was added to a solution of 3 in methanol (0.32 M; CH 3 OH:HCl 29 / 1 v / v), and the reaction mixture was stirred at rt. After 1 hour, the reaction mixture was quenched with saturated sodium bicarbonate solution and extracted with chloroform. The organic layer was dried over sodium sulfate, filtered, and concentrated under vacuum. The crude product was purified by silica gel chromatography (0 - 12% methanol in chloroform) to give 4 as a colorless oil in 79% yield. DCC (2.1 eq) and DMAP (0.05 eq) were added to CH 2 Cl 2Added to a cold (0 °C) solution of 4 in (0.29 M) and palmitic acid (2.1 eq), and the reaction mixture was stirred at 0 °C. After 30 minutes, the reaction mixture was warmed to rt. After 15 hours at rt, additional palmitic acid (0.5 eq) and DCC (0.5 eq) were added and stirring was continued for 24 hours. The reaction mixture was filtered and the crude product was partially purified by silica gel chromatography (0 - 50% ethyl acetate in heptane). Anhydrous CH prepared under nitrogen 2 Cl 2 A solution of the partially purified crude product in (0.17 M) and trimethoxybenzene (0.5 eq) was cannulated into a solution of silver(V) hexafluoroantimonate (0.05 eq) in anhydrous CH 2 Cl 2 (0.09 M). After 2 days at rt, additional silver(V) hexafluoroantimonate (0.05 eq) and trimethoxybenzene (0.5 eq) were added and the reaction mixture was stirred at 50 °C. After 24 hours, the reaction mixture was concentrated under vacuum and purified by silica gel chromatography (10 - 70% ethyl acetate in heptane) to give 5 as a white solid in 89% yield. 1 H NMR (CDCl 3 , 400 MHz), δ 5.23 (m, 1H), 4.34 (dd, 1H), 4.16 (dd, 1H), 3.50 - 3.74 (m, 14H), 2.49 (t, 1H), 2.30 (dd, 4H), 1.60 (m, 4H), 1.25 (m, 48H), 0.88 (t, 6H).

[0190] B. Synthesis of Compound UM - 1007 Oxoadenine UM - 1007 was prepared by the phospholipidation of core oxoadenine 8 with a solution of phosphoramidite 7 generated in - situ from 5 as shown below, using a previously developed tandem method (Tetrahedron Lett., 2016, 57, 2063 - 2066).

Chemical Structure

[0191] Commercially available phosphoramidite 6 (1.3 eq) was added under nitrogen to a solution of dipalmitoyl pegylated glycerol 5 (1.3 eq) in anhydrous methylene chloride (0.12 M), followed by slow addition of tetrazole (1.5 eq). After stirring for 1 h at rt, the reaction mixture was cooled to 0 °C and oxoadenine 8 (1.0 eq) and imidazolium triflate (2.0 eq) were added. The reaction mixture was stirred at 0 °C for 10 min and then warmed to rt. After 1 h, the reaction mixture was quenched by addition of saturated sodium bicarbonate solution. The organic layer was dried over sodium sulfate, concentrated, and crude phosphite was afforded. t-Butyl hydroperoxide (5.5 M in nonane, 2.0 eq) was added to a solution of the crude phosphite (0.12 M) in anhydrous methylene chloride. After 30 min at rt, the reaction mixture was concentrated under vacuum, dissolved in acetonitrile / triethylamine (2.8:1 v / v, 0.05 M), and the reaction mixture was stirred overnight at rt. After concentration under vacuum, the reaction mixture was purified by silica gel chromatography (20 - 100% methanol / acetonitrile (50 / 50) in chloroform) to give UM-1007 in 36% overall yield. 1 H NMR(400MHz,CDCl 3 / CD 3 OD)δ 5.20(m,1H),5.12(m,1H),4.35(dd,1H),4.12-4.17(m,3H),4.05(q,2H),3.79(br s,2H),3.59-3.70(m,12H),3.26(br s,2H),2.80(br s,1H),2.32(q,4H),1.95(m,2H),1.77(m,2H),1.58(m,4H),1.42(m,2H),1.26(m,51H),0.88(m,9H). 13 C NMR(100MHz,CDCl 3 / CD 3OD) δ 174.2, 173.9, 160.7, 153.9, 149.9, 148.5, 106.4, 98.8, 77.8, 73.2, 71.1, 70.9, 70.8, 70.7, 70.5, 70.3, 69.7, 65.4, 65.3, 63.1, 58.8, 53.3, 47.0, 38.6, 34.6, 34.4, 32.2, 29.9, 29.8, 29.6, 29.4, 29.3, 27.4, 25.2, 25.1, 22.9, 20.0, 19.0, 18.9, 14.2, 14.1, 8.8. HRMS [M+H] + Calculated value 1141.7868, measured value 1141.7906.

[0192] Example 2. Biological activity A. In vitro activity of compound UM-1007 IFNα and IL-12p70 responses were measured in primary human peripheral blood mononuclear cells (PBMC) after exposure to various concentrations of oxoadenine compounds.

[0193] Liposomes were prepared using the thin-film method: Phospholipids (DOPC and cholesterol at 40 and 10 mg / mL, respectively) and an agonist (TLR7 / 8 agonist fixed at 2 mg / mL) were dissolved in chloroform at the desired concentration in a round-bottom flask. The solvent was removed under vacuum by rotary evaporation at 45 °C, and the resulting dried thin film was placed in a vacuum chamber overnight to remove any residual solvent. Rehydration buffer was added to the dried thin film, and the suspension was sonicated to resuspend the phospholipids and form unilamellar vesicles. Sonication was continued until the particle size reached <200 nm. Once <200 nm, the formulation was sterile filtered through a 0.22 μm PVDF filter. The concentration of the agonist in the formulation was determined by RP-HPLC using a 5-point calibration curve. Human whole blood was collected from normal healthy donors at the University of Montana (Missoula, MT) using a protocol approved by the Institutional Review Board. Peripheral blood mononuclear cells (PBMCs) were isolated via Ficoll Hypaque 1.077 gradient separation and cultured in 96-well tissue culture plates with RPMI-1640 medium (HyCone™, Logan, UT), Pen / Strep / glutamine (HyCone™, Logan, UT), and 10% heat-inactivated FBS (Corning, Manassas, VA) at 0.5×10 6 cells / well. Human PBMCs were stimulated with increasing concentrations of the indicated compound for 24 hours. Culture supernatants were analyzed for levels of IL-12p70 and IFNα using the Human IL-12p70 DuoSet® ELISA Kit (R&D Systems, Minneapolis, MN) and the Human IFNα VeriKine ELISA Kit (Pestka Biomedical Laboratories, Inc., Piscataway, NJ).

[0194] Pegylated oxoadenine UM-1007, when tested alone or in combination with the TLR4 agonist CRX-601, was a more potent IFNα inducer in human PBMCs than the corresponding non-pegylated oxoadenine (Compound B) (see Figure 1 for structure), indicating that the introduction of the pegylated linker increased IFNα induction (Figure 2A).

[0195] Non-pegylated oxoadenine (Compound B) induced IL-12p70 by itself at high doses (>10 μm), but pegylated oxoadenine UM-1007, when combined with the TLR4 agonist CRX-601, was a much more potent IL-12p70 inducer (125-fold) in human PBMCs than the corresponding non-pegylated oxoadenine (Compound B), indicating that the introduction of the pegylated linker increased the synergistic effect with the TLR4 agonist for IL-12p70 induction (Figure 2B).

[0196] B. In Vivo Activity of Compound UM-1007 The mouse tests were performed in an OLAW- and AAALAC-accredited animal facility according to the University of Montana IACUC guidelines for the care and use of laboratory animals. Balb / c mice (10 mice / group) were immunized intramuscularly twice on days 0 and 14 with increasing doses of adjuvant (three different doses) and monovalent (A / Victoria, H3N2) split influenza antigen (0.15 μg). Serum was collected 14 days after the first immunization (14Dp1) and 14 days after the second immunization (14Dp2) for antibody analysis by ELISA. ELISA plates were coated with 100 μL of 1 μg / mL surfactant-split A / Victoria influenza vaccine. After washing (PBS plus tween 20) and blocking (SuperBlock, Scytek Laboratories), the plates were incubated with diluted serum for 1 hour, followed by incubation with anti-mouse IgG, IgG1 or IgG2a-HRP secondary antibody (Bethyl Laboratories) and TMB substrate (BD). The plates were read at 450 nm. Antibody titers were determined by calculating the titer of each sample at OD0.3.

[0197] The pig study was conducted by PAIRimmune (Laval, Canada), a contract research laboratory. Serum from Yucatan minipigs (5 pigs / group) was collected on days -21 and -1 to evaluate pre-existing anti-influenza serum antibody titers. These pigs exhibited some low level of pre-existing anti-influenza serum antibody titers. Pigs were immunized intramuscularly (500 μL / dose) twice on days 0 and 21 with adjuvant (two different doses) and split influenza antigen (1 / 10 of the human dose). Serum was collected 14 days after the first immunization (14Dp1) and 14 days after the second immunization (14Dp2) for antibody analysis. ELISA plates were coated with 100 μL of 1 μg / mL surfactant-split A / Victoria influenza vaccine. After washing (PBS plus tween 20) and blocking (SuperBlock, Scytek Laboratories), plates were incubated with diluted serum for 1 hour, followed by incubation with anti-pig IgX-HRP conjugate secondary antibody and TMB substrate. Plates were read at 450 nm. Antibody titers were determined by calculating the titer of each sample at OD0.3.

[0198] In the split-influenza mouse study, UM-1007 was formulated alone in liposomes or encapsulated with the TLR4 agonist CRX-601 at a fixed TLR7 / 8:TLR4 ratio of 10:1. Potent anti-influenza IgG titers were detected 14 days after a single intramuscular vaccination in naive mice (14dp1, Figure 3A). Titers were further enhanced by administration of a second booster vaccination (14dp2, Figure 3B). UM-1007 enhanced titers in a dose-dependent manner at 10 μg and showed a 17-fold higher titer compared to antigen-only vaccinated animals. When encapsulated with CRX-601, this response was >25-fold further boosted compared to the antigen-only group.

[0199] UM-1007 was evaluated as an influenza vaccine adjuvant in Yucatan minipigs. UM-1007 was formulated alone in liposomes or encapsulated together with the TLR4 agonist CRX-601. A mixture of UM-1007 liposomes and CRX-601 liposomes was also evaluated. The pigs used in this experiment had some existing anti-influenza serum antibody titers. Regardless, all TLR7 / 8-TLR4 agonist adjuvant groups demonstrated higher mean antibody titers than the antigen-only group after single (not shown) or booster vaccination (14 dp2, Figure 4). Increases in influenza-specific IgG1 (Figure 4A) and IgG2 (Figure 4B) were clearly demonstrated for UM-1007 and CRX-601 encapsulated together, compared to CRX-601 or UM-1007 alone.

[0200] C. Effect of acyl side chains on the in vitro activity of selected TLR agonists Compound B has an oleoyl acyl chain, while UM-1007 has a palmitoyl acyl chain (Figure 1). The increase in in vitro activity observed above for UM-1007 versus Compound B was not due to different acyl chains, as shown when comparing the activity of palmitoyloxoadenine Compound C to oleoyloxoadenine Compound A (Figure 5). Both oxoadenines induced similar levels of TNFα (Figure 5A), IFNα (Figure 5B) (a slight increase in activity was observed for palmitoyloxoadenine), and IL-12p70 (Figure 5C).

[0201] Example 3. Biological Activity A. In vivo activity of compound UM-1007 Young female Balb / c mice (approximately 8 weeks old) were subcutaneously implanted in the right flank with 1×10 6 CT26 cells (a mouse colon cancer cell line, syngeneic in Balb / c mice). When the tumors reached 75 mm 3When the average size was reached (11 days after transplantation of CT26 cells), the mice were randomly assigned to treatment groups. Starting on day 12, 10 μg or 50 μg of UM-1007 in an aqueous formulation (2% glycerol in sterile water) was injected IV once a week for 2 weeks. As shown in FIGS. 6A and 6B, this resulted in slow tumor growth compared to untreated controls.

[0202] Example 4. Adsorption of UM1007 to Alum Materials: Alhydrogel® (10 mg / mL aluminum) is an aluminum hydroxide wet gel suspension available from InvivoGen. Alhydrogel® particles have a net positive charge suitable for adsorption of negatively charged antigens (e.g., antigens having an isoelectric point below the pH of the formulation) at pH 5 - 7.

[0203] Adju-Phos® (0.5% aluminum) is an aluminum phosphate wet gel suspension available from InvivoGen. Adju-Phos® particles have a negative charge suitable for adsorption of positively charged antigens (e.g., antigens having an isoelectric point above the pH of the formulation) at pH 5 - 7.

[0204] Methods: Preparation of an aqueous formulation of UM 1007: UM 1007 was salt-treated using choline bicarbonate by a platform drying salt procedure. Briefly, UM 1007 was placed in a glass vial, an appropriate amount of tetrahydrofuran and 0.8 equivalents of choline bicarbonate were added to obtain a clear solution. The mixture was vortexed thoroughly and the solvent was evaporated under reduced pressure using a rotary evaporator. The thin film formed on the walls of the glass vial was rehydrated with 2% glycerol and sonicated using a sonicator at <35 °C for 210 minutes to reduce the particle size. The concentration of UM 1007 was estimated using the RP-HPLC method.

[0205] Adsorption of UM 1007 to Alhydrogel and Adju-Phos Adsorption experiments of UM 1007 on Alhydrogel and Adju-Phos were carried out at two different weight ratios of UM 1007 to aluminum (1:1 and 1:2 w / w) and in three different solutions (2% glycerol, water for washing (WIFI), and TRIS buffer (pH 8.1). A series of UM 1007-alum formulations were prepared at room temperature by mixing different amounts of the aqueous UM1007 formulation, each of the storage buffers (2% glycerol, water for washing, and pH 8.1 TRIS buffer), and the aluminum source (1 mg / mL Alhydrogel or Adju-Phos). Alhydrogel and Adju-Phos were diluted to 1 mg / mL before the experiment. Different volumes of UM 1007 (0.7215 mg / mL) and alum (1 mg / mL) were gently mixed by end-over-end rotation at room temperature for about 1 hour. The samples were centrifuged at 4,000 rpm for 5 minutes, and the supernatant was analyzed using RP-HPLC to determine the amount of UM 1007 that was not adsorbed. The percentage of UM 1007 adsorbed to alum was estimated by comparing with the amount of UM 1007 in the control UM 1007. The results of the tests are shown in Table 1.

[0206]

Table 1

[0207] For reasons of completeness, various aspects of the present invention are described in the numbered clauses below.

[0208] Clause 1. Formula (I)

Chemical formula

[0209] Clause 2. R 2 is hydrogen or C(O)R 4 of the compound of Clause 1 or a pharmaceutically acceptable salt thereof.

[0210] Clause 3. R 3 is C(O)R 4 of the compound of Clause 1 or Clause 2 or a pharmaceutically acceptable salt thereof.

[0211] Clause 4. R 4 is, in each occurrence, (CH 2 ) 10 CH 3 , (CH 2 ) 12 CH 3 , (CH 2 ) 14 CH 3 , (CH 2 ) 16 CH 3 and (CH 2 ) 7 CH = CH(CH2 ) 7 CH 3 A compound according to any one of clauses 1 to 3, or a pharmaceutically acceptable salt thereof, independently selected from

[0212] Clause 5.R 4 is (CH 2 ) 14 CH 3 A compound according to any one of clauses 1 to 4, or a pharmaceutically acceptable salt thereof.

[0213] Clause 6. n is 1. A compound according to any one of clauses 1 to 5, or a pharmaceutically acceptable salt thereof.

[0214] Clause 7. m is 2. A compound according to clauses 1 to 6, or a pharmaceutically acceptable salt thereof.

[0215] Clause 8. Z is (C 2 alkylene - O) q A compound according to any one of clauses 1 to 7, or a pharmaceutically acceptable salt thereof.

[0216] Clause 9. q is 3, 6, 9, 12 or 16. A compound according to any one of clauses 1 to 8, or a pharmaceutically acceptable salt thereof.

[0217] Clause 10. q is 3. A compound according to any one of clauses 1 to 9, or a pharmaceutically acceptable salt thereof.

[0218] Clause 11. Formula (Ia):

Chemical formula

[0219] Clause 12.

Chemical formula

Chemical formula

[0220] A compound of any one of clauses 1 to 12, wherein the pharmaceutically acceptable salt is a choline salt.

[0221] A compound of any one of clauses 1 to 13 or a pharmaceutically acceptable salt thereof, wherein the compound is a TLR7 antagonist.

[0222] A compound of any one of clauses 1 to 13 or a pharmaceutically acceptable salt thereof, wherein the compound is a TLR8 antagonist.

[0223] A formulation comprising microparticles or nanoparticles comprising a compound of any one of clauses 1 to 15 or a pharmaceutically acceptable salt thereof.

[0224] The formulation of clause 16, wherein the microparticles or nanoparticles comprise liposomes, micelles, polymeric particles, block copolymers, silica particles, emulsions or combinations thereof.

[0225] An adjuvant composition comprising an effective amount of a compound of any one of clauses 1 to 15 or a pharmaceutically acceptable salt thereof.

[0226] The adjuvant composition of clause 18, wherein the adjuvant composition induces a Th1-type immune response.

[0227] A method of inducing enhancement of an immune response in a subject, the method comprising administering to the subject an immunogenic composition comprising a compound of any one of clauses 1 to 15, a pharmaceutically acceptable salt thereof or the adjuvant composition of clause 18 or clause 19.

[0228] Article 21. A vaccine composition comprising an antigen and any one of the compounds of Articles 1 to 15, a pharmaceutically acceptable salt thereof, or an adjuvant composition of Article 18 or Article 19.

[0229] Article 22. The vaccine composition of Article 21, wherein the antigen is derived from bacteria, viruses, bacteriophages, fungi, prions, neoplasms, autoantigens, animals, plants, recombinants or synthetic substances.

[0230] Article 23. The vaccine composition of Article 21 or 22, wherein the antigen is in the form of a peptide or polypeptide.

[0231] Article 24. The vaccine composition of Article 21 or 22, wherein the antigen is in the form of a hapten or a hapten conjugated to a carrier protein.

[0232] Article 25. The vaccine composition of Article 21 or 22, wherein the antigen is an allergen.

[0233] Article 26. A method of inducing or enhancing the immunogenicity of an antigen in a subject, the method comprising administering to the subject any one of the vaccine compositions of Articles 21 to 25, a pharmaceutically acceptable salt thereof, or an adjuvant composition of Articles 18 to 19.

[0234] Article 27. A pharmaceutical composition comprising an effective amount of any one of the compounds of Articles 1 to 15 or a pharmaceutically acceptable salt thereof.

[0235] Article 28. The pharmaceutical composition of Article 27, further comprising an additional therapeutic agent.

[0236] Article 29. The pharmaceutical composition of Article 28, wherein the additional therapeutic agent is an adjuvant, an immunostimulant, a chemotherapeutic agent, an immunomodulator or a combination thereof.

[0237] Article 30. The pharmaceutical composition of Article 29, wherein the adjuvant is a TLR4 ligand.

[0238] Clause 31. The pharmaceutical composition of Clause 29, wherein the adjuvant is an aluminum salt.

[0239] Clause 32. The pharmaceutical composition of Clause 31, wherein any one of the compounds of Claims 1 to 15 is adsorbed to an aluminum salt.

[0240] Clause 33. The pharmaceutical composition of Clause 32, further comprising an antigen adsorbed to an aluminum salt together with the compound.

[0241] Clause 34. The pharmaceutical composition of Clause 29, wherein the immunomodulator is an immune checkpoint inhibitor, a tumor phagocytosis inducer, or a combination thereof.

[0242] Clause 35. A method of modulating an immune response in a subject, the method comprising administering to the subject any one of the pharmaceutical compositions of Clauses 27 to 34.

[0243] Clause 36. The method of Clause 35, wherein the immune response of the subject is increased.

[0244] Clause 37. The method of Clause 35 or 36, wherein the subject is suffering from cancer, an autoimmune disorder, an allergy, or an infectious disease.

[0245] Clause 38. The method of Clause 37, wherein the infectious disease is a viral, bacterial, or prion infection.

[0246] Clause 39. A method of treating, preventing, or reducing the susceptibility to a disease or disorder in a subject, the method comprising administering to the subject in need thereof a therapeutically effective amount of any one of the compounds of Claims 1 to 15, a pharmaceutically acceptable salt thereof, or any one of the pharmaceutical compositions of Clauses 27 to 34.

[0247] Clause 40. The method of Clause 39, wherein the disease or disorder is an allergy, an autoimmune disease or disorder, an infectious disease or infection, or cancer.

[0248] Clause 41. An infectious disease or an infectious disorder is a method of Clause 40 caused by a viral, bacterial or prion infection. Aspects of the Invention [Aspect 1] Formula (I) [Chemical Formula] (In the formula, R 1 is -CH(CH 3 )(CH 2 ) 2 CH 3 ; R 2 is C(O)R 4 , C 6 ~C 20 alkyl or C 6 ~C 20 alkenyl; R 3 is C(O)R 4 , C 6 ~C 20 alkyl or C 6 ~C 20 alkenyl; R 4 is, in each occurrence, independently selected from C 15 ~C 6 alkyl, C 17 ~C 16 alkyl and C 17 ~C 6 alkenyl; n is 1; m is 2; Z is (C 2 alkylene - O) q ; and q is 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19 or 20) of the compound or a pharmaceutically acceptable salt thereof. [Aspect 2] The compound or a pharmaceutically acceptable salt thereof according to Aspect 1, wherein R 2 is C(O)R 4 . [Aspect 3] The compound or a pharmaceutically acceptable salt thereof according to Aspect 1, wherein R 3 is C(O)R 4The compound according to embodiment 2 or a pharmaceutically acceptable salt thereof. [Embodiment 4] R 4 is, in each occurrence, (CH 2 ) 14 CH 3 , (CH 2 ) 10 CH 3 , (CH 2 ) 12 CH 3 , (CH 2 ) 16 CH 3 and (CH 2 ) 7 CH=CH(CH 2 ) 7 CH 3 independently selected from the compounds according to any one of embodiments 1 to 3 or a pharmaceutically acceptable salt thereof. [Embodiment 5] R 4 is (CH 2 ) 14 CH 3 the compound according to embodiment 4 or a pharmaceutically acceptable salt thereof. [Embodiment 6] q is 3, 6, 9, 12 or 16, the compound according to embodiment 4 or a pharmaceutically acceptable salt thereof. [Embodiment 7] q is 3, the compound according to embodiment 6 or a pharmaceutically acceptable salt thereof. [Embodiment 8] Formula (Ia):

Chemical formula

Chemical formula

Chemical formula

Chemical formula

Claims

1. Formula (I) 【Chemical Formula 1】 (wherein R １ is -CH(CH ３ )(CH ２ ) ２ CH ３ and; R ２ is C(O)R ４ C ６ ~C ２０ alkyl or C ６ ~C ２０ alkenyl; R ３ is C(O)R ４ 、C ６ ~C ２０ alkyl or C ６ ~C ２０ is alkenyl; R ４ is, in each occurrence, independently selected from C ６ to C １７ alkyl and C ６ to C １７ alkenyl; n is 1; and m is 2) and a pharmaceutically acceptable salt thereof.

2. R ２ is C(O)R ４ The compound according to claim 1 or a pharmaceutically acceptable salt thereof.

3. R ３ is C(O)R ４ The compound according to claim 2 or a pharmaceutically acceptable salt thereof.

4. R ４ is, in each occurrence, (CH ２ ), １４ CH ３ , (CH ２ ), １０ CH ３ , (CH ２ ), １２ CH ３ , (CH ２ ), １６ CH ３ and (CH ２ ), ７ CH=CH(CH ２ ), ７ CH ３ independently selected from, a compound according to any one of claims 1 to 3 or a pharmaceutically acceptable salt thereof.

5. R ４ is (CH ２ ) １４ CH ３ The compound according to claim 4 or a pharmaceutically acceptable salt thereof.

6. Formula (Ia): [Chemical Formula 2] The compound according to claim 1, which is a compound or a pharmaceutically acceptable salt thereof.

7. 【Fig. 3】 【Chemical Formula 4】 The compound according to claim 1 or a pharmaceutically acceptable salt thereof, which is selected from the group consisting of

8. 【Fig. 5】 The compound according to claim 7 or a pharmaceutically acceptable salt thereof, which is

9. The compound according to claim 4, wherein the pharmaceutically acceptable salt is a choline salt.

10. A formulation comprising microparticles or nanoparticles containing the compound according to claim 1 or a pharmaceutically acceptable salt thereof.

11. The formulation according to claim 10, wherein the microparticles or nanoparticles comprise liposomes, micelles, polymer particles, block copolymers, silica particles, emulsions or combinations thereof.

12. A vaccine composition comprising an antigen and the compound according to claim 1 or a pharmaceutically acceptable salt thereof.

13. A pharmaceutical composition comprising the compound according to claim 1 or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier.

14. The pharmaceutical composition according to claim 13, further comprising a TLR4 ligand.

15. The compound is [Chemical Formula 6] and The TLR4 ligand is 【Chemical Formula 7】 or a pharmaceutically acceptable salt thereof, the pharmaceutical composition according to claim 14.

16. The pharmaceutical composition according to claim 13, further comprising an aluminum salt.

17. The pharmaceutical composition according to claim 16, wherein the compound or the pharmaceutically acceptable salt thereof is adsorbed to the aluminum salt.

18. The pharmaceutical composition according to claim 17, further comprising an antigen adsorbed to the aluminum salt together with the compound or the pharmaceutically acceptable salt thereof.

19. A pharmaceutical composition for use in a method of modulating an immune response in a subject, comprising the compound according to claim 1 or a pharmaceutically acceptable salt thereof.

20. The pharmaceutical composition according to claim 19, wherein the immune response in the subject is increased.

21. The pharmaceutical composition according to claim 19, wherein the subject is suffering from cancer or an infectious disease.

22. The pharmaceutical composition according to claim 20, for use in combination with an antigen.

23. The pharmaceutical composition according to claim 19, for use in combination with a TLR4 ligand.

24. The TLR4 ligand is 【Chemical 8】 The pharmaceutical composition according to claim 23, which is or a pharmaceutically acceptable salt thereof.

25. The pharmaceutical composition according to claim 19, for use in combination with an aluminum salt adjuvant.

26. The pharmaceutical composition according to claim 25, wherein the compound or the pharmaceutically acceptable salt thereof is adsorbed on the aluminum salt adjuvant.

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