Formulated and / or co-formulated liposome compositions containing IDO antagonist prodrugs useful in treatment of cancer and methods thereof

Abstract

To provide formulated and / or co-formulated liposome compositions containing IDO antagonist prodrugs useful in treatment of cancer and methods thereof.SOLUTION: Formulated and / or co-formulated liposomes comprising IDO prodrugs and methods of making the liposomes are disclosed herein. The IDO prodrug compositions comprise a drug moiety, a lipid moiety, and linkage unit that inhibit IDO-1. The IDO prodrugs can be formulated and / or co-formulated into a liposome to provide a method of treating cancer, immunological disorders, and other disease by utilizing a targeted drug delivery vehicle.SELECTED DRAWING: None

Description

Technical Field

[0001] Cross - reference to Related Applications This application claims priority to U.S. Provisional Patent Application No. 62 / 974,086, filed on November 12, 2019, the content of which is hereby incorporated by reference in its entirety.

[0002] Statement of Rights to Inventions Made Under Federally Sponsored Research Not applicable.

[0003] Field of the Invention The invention described herein relates to prodrug compositions that inhibit the indoleamine - pyrrole 2,3 - dioxygenase (IDO) enzyme after release of an active inhibitor from the prodrug and to nanopharmaceuticals containing such prodrugs. Specifically, the invention relates to prodrug compositions formulated within a nanocarrier (e.g., liposome) and used as a vehicle for cancer treatment in humans. The invention further relates to the treatment of cancer as well as other immune disorders and diseases.

Background Art

[0004] Background of the Invention Cancer is the second leading cause of death worldwide after coronary artery disease. Every year, millions of people die from cancer, and in the United States alone, well over 500,000 people die from cancer each year. In 2017, 1,688,780 new cancer cases were diagnosed (American Cancer Society). Deaths due to heart disease have decreased significantly, while deaths due to cancer generally have increased. It is predicted that cancer will be the leading cause of death early in the next century unless medical developments change current trends.

[0005] Several cancers are noted for having high mortality rates. In particular, cancers of the lung (18.4% of all cancer deaths), breast (6.6% of all cancer deaths), colorectum (9.2% of all cancer deaths), liver (8.2% of all cancer deaths) and stomach (8.2% of all cancer deaths) are the leading causes of cancer death worldwide in both men and women of all ages (GLOBOCAN 2018). These cancers, and substantially all other cancers, share the common lethal feature of metastasizing to sites distant from the primary tumor, and with few exceptions, metastatic disease is lethal. Furthermore, even cancer patients who initially survive their primary cancer, as is common experience, their lives are dramatically changed. Many cancer patients experience strong anxiety by being aware of the possibility of recurrence or treatment failure. Many cancer patients also experience physical debilitation after treatment. Furthermore, many cancer patients experience recurrence of the disease.

[0006] Cancer treatment has improved over the past few decades and survival rates have increased, but due to the heterogeneity of cancer, new treatment strategies that utilize multiple treatment modalities are still needed. This is particularly true for the treatment of solid tumors in anatomically critical sites (e.g., glioblastoma, head and neck squamous cell carcinoma, and lung adenocarcinoma) that may be limited to standard radiotherapy and / or chemotherapy. Nevertheless, the adverse effects of these therapies are chemoresistance and radioresistance, and in addition to severe side effects that reduce the quality of life of patients, they promote local recurrence, distant metastasis, and another primary tumor.

[0007] Indoleamine-pyrrole 2,3-dioxygenase (IDO or INDO) is a heme-containing enzyme encoded by the IDO1 gene in humans. IDO is the first and rate-limiting enzyme in tryptophan catabolism via the kynurenine pathway, and thus can cause depletion of tryptophan, which in turn can slow the growth of microorganisms and T cells. Additionally, IDO is an immune checkpoint molecule in the sense that it is an immunomodulatory enzyme produced by some selectively activated macrophages and other immune regulatory cells (also used as an immune evasion strategy by many tumors and chronic infectious viruses). IDO is known to suppress T cells and NK cells, generate and activate Tregs and myeloid-derived suppressor cells, and promote the growth of new blood vessels that supply tumors (angiogenesis). IDO enables tumor cells to escape from the immune system through depletion of L-tryptophan in the tumor microenvironment and production of the catabolite kynurenine, and selectively impairs the proliferation and survival of T cells. A wide range of human cancers, such as prostate cancer, colorectal cancer, pancreatic cancer, cervical cancer, gastric cancer, ovarian cancer, head cancer, and lung cancer, overexpress human IDO (hIDO). IDO is involved in immune regulation through its ability to limit T cell function and operate mechanisms of immune tolerance. New evidence suggests that IDO is activated during tumorigenesis and helps malignant cells escape eradication by the immune system. MUNN et al., Trends in Immunology, 37(3): pp. 193-207 (March 2016) and PENDERGRAST et al., Cancer Immunol Immunother., 63(7): pp. 721-735 (July 2014).

[0008] Furthermore, a prodrug is an agent or compound that is metabolized (i.e., converted in the body) into a pharmacologically active drug after administration. Instead of administering the drug directly, the corresponding prodrug is used instead to improve the absorption, distribution, metabolism, and / or excretion profile of the drug. Prodrugs are often designed to improve bioavailability, for example, when the drug itself is poorly absorbed from the gastrointestinal tract. Prodrugs can be used to improve the extent to which a drug selectively interacts with cells or processes that are not its intended target. This reduces the harmful or unintended effects of drugs, which is particularly important in treatments such as chemotherapy that can sometimes have severe, unwanted side effects. Thus, prodrugs can be regarded as drugs containing special non-toxic protecting groups that are temporarily used to change or eliminate undesirable properties of the parent molecule.

[0009] Finally, a nanocarrier is a nanomaterial that is used for the transport of another substance such as a drug. There are many different types of nanocarriers. For example, to name a few, nanocarriers include polymer conjugates, polymer nanoparticles, lipid-based carriers, and dendrimers. The different types of nanomaterials used in nanocarriers enable the delivery of both hydrophobic and hydrophilic drugs throughout the body. Since the human body mainly contains water, the ability to effectively deliver hydrophobic drugs to humans is a major therapeutic advantage of nanocarriers. Nanocarriers are promising in the drug delivery process because they can deliver drugs to site-specific targets, delivering the drug to a particular organ or cell and not to other organs or cells. Site-specificity is a major therapeutic advantage as it prevents the drug from being delivered to the wrong place. Furthermore, nanocarriers can be useful in reducing the harmful and more extensive toxicity of chemotherapy to rapidly growing healthy cells in the body, showing particular promise for use in chemotherapy. Since chemotherapeutic drugs can be extremely toxic to human cells, it is important that the chemotherapeutic drug is delivered to the tumor without being released to other parts of the body.

[0010] From the above description, it will be readily apparent to those skilled in the art that a new treatment paradigm is required in the treatment of cancer and other immune diseases. By using a novel prodrug in combination with the latest nanocarrier format, a new disease treatment can be achieved with the overall goals of more effective treatment, reduction of side effects, and greater therapeutic utility in the treatment of cancer, particularly cancer in solid tumors. In view of the current deficiencies associated with cancer treatment, it is an object of the present invention to provide a novel and improved method for treating cancer, immune disorders, and other diseases by utilizing a prodrug encapsulated within a nanocarrier.

Prior Art Documents

Non-Patent Documents

[0011]

Non-Patent Document 1

Non-Patent Document 2

Summary of the Invention

Means for Solving the Problems

[0012] Summary of the Invention The present invention provides an IDO inhibitor prodrug (an "IDO prodrug") composition comprising an IDO inhibitor agent, a lipid, and a biologically cleavable linker. In certain embodiments, the nanocarrier comprising the IDO prodrug is formulated for use as a delivery modality for treating human diseases such as cancer, including solid tumor cancer, and other immune disorders. In certain embodiments, the nanocarrier comprises a lipid bilayer (i.e., a liposome) that can be incorporated into a drug delivery vehicle. In a further preferred embodiment, the liposome comprises cholesterol hemisuccinate ("CHEMS"). In a further preferred embodiment, the liposomes of the present invention comprise stearic acid.

[0013] In a further embodiment, the present invention provides a method of delivering an IDO inhibitor to a tumor, comprising: (i) synthesizing an IDO prodrug; (ii) formulating the IDO prodrug of the present invention into a nanocarrier of the present invention; and (iii) administering the nanocarrier to a patient.

[0014] In another embodiment, the present invention provides a method of delivering an IDO inhibitor to a tumor together with one or more additional immunomodulatory agents, comprising: (i) synthesizing an IDO prodrug; (ii) co-formulating the IDO prodrug of the present invention with one or more additional immunomodulatory agents of the present invention into a nanocarrier; and (iii) administering the nanocarrier to a patient.

[0015] In another embodiment, the immunomodulatory agent comprises an immunogenic cell death-inducing chemotherapeutic agent, a PD-1 agonist, a toll receptor agonist, a STING agonist, a CTLA4 inhibitor, and / or prodrugs thereof.

[0016] In another embodiment, the present disclosure teaches a method of synthesizing an IDO prodrug.

[0017] In another embodiment, the present disclosure teaches a method of formulating an IDO prodrug within a nanocarrier including, but not limited to, liposomes.

[0018] In another embodiment, the present disclosure teaches a method of treating cancer, immune disorders and other diseases in humans using the nanocarriers of the present disclosure. BRIEF DESCRIPTION OF THE DRAWINGS

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Mode for Carrying Out the Invention

[0046] Detailed Description of the Invention Summary of Sections I.) Definitions II.) Prodrugs III.) Chemical Compounds IV.) Lipids V.) Linking Unit (“LU”) VI.) Nanocarriers VII.) Liposomes VIII.) Pharmaceutical Formulations IX.) Combination Therapies X.) Methods of Delivering Liposomes Containing Prodrugs to Cells XI.) Methods of Treating Cancer and Other Immune Disorders XII.) Kits / Articles of Manufacture I.) Definitions:

[0047] Unless otherwise defined, all technical terms, notations, and other scientific or technical terms or phrases used herein are intended to have the meaning commonly understood by one of ordinary skill in the art, unless the context clearly indicates otherwise. In some cases, terms having commonly understood meanings are defined herein for clarity and / or ease of reference, and inclusion of such definitions herein should not necessarily be construed as representing a substantial difference from what is commonly understood in the art.

[0048] When a trade name is used herein, reference to the trade name refers to the product formulation, generic pharmaceutical, and pharmaceutically active ingredient of the product of that trade name, unless the context otherwise indicates.

[0049] As used herein, the term “about,” when referring to a value or amount of size (i.e., diameter), weight, concentration, or percentage, is intended to encompass variations of, for example, ±20% or ±10% from the specified amount, in one example, ±5% in another example, ±1% in another example, and ±0.1% in yet another example, as appropriate to carry out the disclosed method.

[0050] As used herein, the term "and / or" when used in the context of a list of entities refers to the entities being present individually or in combination. Thus, for example, the phrase "A, B, C, and / or D" includes A, B, C, and D individually, but also includes any and all combinations and sub - combinations of A, B, C, and D.

[0051] As used herein, numerical ranges described by endpoints include all numbers and fractions subsumed within that range (e.g., 1 - 5 includes, but is not limited to, 1, 1.5, 2, 2.75, 3, 3.90, 4, and 5).

[0052] As used herein, the phrase "consisting essentially of" limits the claims to the specified materials or steps, plus those that do not substantially affect the basic and novel characteristics of the claimed subject matter.

[0053] The terms "advanced cancer", "locally advanced cancer", "advanced disease", and "locally advanced disease" mean cancer that has spread through the associated tissue membrane, and include stage C disease under the American Urological Association (AUA) system, stage C1 - C2 disease under the Whitmore - Jewett system, and stage T3 - T4 and N + disease under the TNM (tumor, node, metastasis) system. Generally, surgery is not recommended for patients with locally advanced disease, and these patients have a substantially worse prognosis compared to patients with clinically localized (organ - confined) cancer.

[0054] As used herein, the term "alkyl" includes, for example, methyl, ethyl, propyl, isopropyl, butyl, isobutyl, tert - butyl, pentyl, hexyl, octyl, ethenyl, propenyl, butenyl, pentenyl, hexenyl, octenyl, butadienyl, propynyl, butynyl, pentynyl, hexynyl, heptynyl, heptynyl, and arylenyl groups, C1 - C 20can refer to a linear (i.e., "straight-chain"), branched or cyclic, saturated or at least partially unsaturated, and optionally unsaturated (i.e., alkenyl and alkynyl) hydrocarbon chain (including both ends). "Branched" refers to an alkyl group in which a lower alkyl group such as methyl, ethyl or propyl is attached to a straight-chain alkyl chain. "Lower alkyl" refers to an alkyl group having 1 to about 8 carbon atoms (i.e., C1-C8 alkyl), for example, an alkyl group having 1, 2, 3, 4, 5, 6, 7 or 8 carbon atoms. "Higher alkyl" refers to an alkyl group having about 10 to about 20 carbon atoms, for example, an alkyl group having 10, 11, 12, 13, 14, 15, 16, 17, 18, 19 or 20 carbon atoms. In certain embodiments, "alkyl" particularly refers to C1-C8 straight-chain alkyl. In other embodiments, "alkyl" particularly refers to C 1~8 branched-chain alkyl.

[0055] The alkyl group can be optionally substituted with one or more alkyl group substituents which can be the same or different ("substituted alkyl"). The term "alkyl group substituent" includes, but is not limited to, alkyl, substituted alkyl, halo, arylamino, acyl, hydroxyl, aryloxyl, alkoxyl, alkylthio, arylthio, aralkyloxyl, aralkylthio, carboxyl, alkoxycarbonyl, oxo and cycloalkyl. In some embodiments, one or more oxygen, sulfur or substituted or unsubstituted nitrogen atoms can be inserted along the alkyl chain as needed, and the nitrogen substituents are hydrogen, lower alkyl (also referred to herein as "alkylaminoalkyl") or aryl.

[0056] Accordingly, as used herein, the term "substituted alkyl" includes an alkyl group in which one or more atoms or functional groups of the alkyl group are replaced with another atom or functional group including, for example, alkyl, substituted alkyl, halogen, aryl, substituted aryl, alkoxyl, hydroxyl, nitro, amino, alkylamino, dialkylamino, sulfate and mercapto, as defined herein.

[0057] As used herein, the term "aryl" refers to an aromatic substituent that can be a single aromatic ring, or a plurality of aromatic rings that are fused together, covalently bonded, or bonded to a common group such as a methylene or ethylene moiety, but not limited thereto. The common linking group can also be a carbonyl as in benzophenone, or an oxygen as in diphenyl ether, or a nitrogen as in diphenylamine. The term "aryl" specifically encompasses heteroaromatic compounds. The aromatic ring can include, inter alia, phenyl, naphthyl, biphenyl, diphenyl ether, diphenylamine, and benzophenone. In certain embodiments, the term "aryl" means a cyclic aromatic containing from about 5 to about 10 carbon atoms, such as 5, 6, 7, 8, 9, or 10 carbon atoms, including 5- and 6-membered aromatic and heteroaromatic rings. An aryl group can be optionally substituted with one or more aryl group substituents which can be the same or different ("substituted aryl"), and "aryl group substituents" include alkyl, substituted alkyl, aryl, substituted aryl, aralkyl, hydroxyl, alkoxyl, aryloxyl, aralkyloxyl, carboxyl, acyl, halo, nitro, alkoxycarbonyl, aryloxycarbonyl, aralkoxycarbonyl, acyloxyl, acylamino, aroylamino, carbamoyl, alkylcarbamoyl, dialkylcarbamoyl, arylthio, alkylthio, alkylene, and -NR’R’’, where R’ and R’’ can each independently be hydrogen, alkyl, substituted alkyl, aryl, substituted aryl, and aralkyl. Specific examples of aryl groups include, but are not limited to, cyclopentadienyl, phenyl, furan, thiophene, pyrrole, pyran, pyridine, imidazole, benzimidazole, isothiazole, isoxazole, pyrazole, pyrazine, triazine, pyrimidine, quinoline, isoquinoline, indole, carbazole, etc.

[0058] As used herein, "heteroaryl" refers to an aryl group containing one or more non-carbon atoms (e.g., O, N, S, Se, etc.) in the backbone of the ring structure. Nitrogen-containing heteroaryl moieties include, but are not limited to, pyridine, imidazole, benzimidazole, pyrazole, pyrazine, triazine, pyrimidine, and the like.

[0059] The terms "anticancer agent", "chemotherapeutic agent", and "anticancer prodrug" refer to drugs (i.e., chemical compounds) or prodrugs that are known or believed to have the potential to treat cancer (i.e., kill cancer cells, prevent the growth of cancer cells, or treat symptoms associated with cancer). In some embodiments, the term "chemotherapeutic agent" as used herein refers to a non-PS molecule that is used to treat cancer and / or has cytotoxic activity. More traditional or conventional chemotherapeutic agents can be described by mechanism of action or by class of chemical compounds and can include, but are not limited to, alkylating agents (e.g., melphalan), anthracyclines (e.g., doxorubicin), cytoskeletal disruptors (e.g., paclitaxel), epothilones, histone deacetylase inhibitors (e.g., vorinostat), inhibitors of topoisomerase I or II (e.g., irinotecan or etoposide), kinase inhibitors (e.g., bortezomib), nucleotide analogs or precursors thereof (e.g., methotrexate), peptide antibiotics (e.g., bleomycin), platinum-based agents (e.g., cisplatin or oxaliplatin), retinoids (e.g., tretinoin), and vinca alkaloids (e.g., vinblastine).

[0060] "Aralkyl" refers to an -alkyl-aryl group in which the alkyl and / or aryl moieties are optionally substituted.

[0061] "Alkylene" refers to a straight-chain or branched divalent aliphatic hydrocarbon group having from 1 to about 20 carbon atoms, for example, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19 or 20 carbon atoms. The alkylene group can be straight-chain, branched or cyclic. The alkylene group can also, optionally, be unsaturated and / or substituted with one or more "alkyl group substituents". One or more oxygen, sulfur or substituted or unsubstituted nitrogen atoms can be inserted along the alkylene group as needed (also referred to herein as "alkylaminoalkyl"), and the nitrogen substituents are the aforementioned alkyl. Exemplary alkylene groups include methylene (-CH2-); ethylene (-CH2-CH2-); propylene (-(CH2)3-); cyclohexylene (-(C6H 10 -); -CH=CH-CH=CH-; -CH=CH-CH2-; -(CH2) q -N(R)-(CH2), -, q are each an integer from 0 to about 20, for example, 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19 or 20, and R is hydrogen or lower alkyl; methylenedioxyl (-O-CH2-O-); and ethylenedioxyl (-O-(CH2)2-O-). The alkylene group can have from about 2 to about 3 carbon atoms and can further have from 6 to 20 carbons.

[0062] The term "arylene" refers to a divalent aromatic group, for example, a divalent phenyl or naphthyl group. The arylene group can, optionally, be substituted with one or more aryl group substituents and / or can contain one or more heteroatoms.

[0063] The term "amino" refers to the group -N(R)2, where each R is independently H, alkyl, substituted alkyl, aryl, substituted aryl, aralkyl or substituted aralkyl. The terms "aminoalkyl" and "alkylamino" can refer to the group -N(R)2, where each R is H, alkyl or substituted alkyl and at least one R is alkyl or substituted alkyl. The terms "arylamine" and "aminoaryl" refer to the group -N(R)2, where each R is H, aryl or substituted aryl and at least one R is aryl or substituted aryl, such as aniline (i.e., -NHC6H5).

[0064] "Bioreactive nanomaterial" refers to engineered biomaterials that induce or catalyze a biological response. In certain embodiments, the nanomaterial induces a response by one or more properties selected from the group consisting of composition, size, shape, aspect ratio, solubility, electronic, redox, surface display, surface coating, hydrophobicity, hydrophilicity, atomically thin nanosheets, or functionalized surface groups, and catalyzes a biological response at various nano / bio interfaces. In certain embodiments, the bioreactive nanomaterial has the ability to inhibit the IDO-1 biological response in cells (e.g., in tumor cells) and / or the ability to activate the innate immune system through the delivery of "danger signals" and adjuvant effects.

[0065] "Bulk" (also known as the drug substance) means the drug substance or pharmaceutical product that is not filled in the final container for distribution. Final formulated bulk generally refers to the pharmaceutical product that has been formulated and stored or held prior to filling. The drug substance can be stored or held as "bulk" or "concentrated bulk" prior to formulation into a pharmaceutical product.

[0066] The terms "carboxylate" and "carboxylic acid" can refer to the groups -C(=O)O - and -C(=O)OH, respectively. The term "carboxyl" can also refer to the -C(=O)OH group.

[0067] As used herein, the terms "conjugate" and "conjugated" can refer to the attachment (e.g., covalent attachment) of two or more components (e.g., chemical compounds, polymers, biomolecules, particles, etc.) to each other. In some embodiments, a conjugate can include a monovalent moiety derived from two different chemical compounds covalently attached via a divalent linker moiety (e.g., alkylene or arylene optionally substituted). In some embodiments, the linker can contain one or more biodegradable linkages such that one or more linkages in the linker can be cleaved when the prodrug is exposed to a particular physiological environment or enzyme (e.g., esterase).

[0068] The term "compound" refers to the chemical compound (e.g., prodrug) itself and includes, without limitation and unless the context clearly indicates otherwise, the following, whether or not explicitly stated: amorphous and crystalline forms of the compound, including polymorphic forms, which may be part of a mixture or in isolated form; free acid and free base forms of the compound, typically in the form shown in the structures provided herein; isomers of the compound, including optical isomers and tautomers, where optical isomers include enantiomers and diastereomers, chiral and achiral isomers, and optical isomers include mixtures of optical isomers, including racemic and non-racemic mixtures, in addition to isolated optical isomers; isomers may be in isolated form or in mixture with one or more other isomers; isotopes of the compound, including deuterium-containing and tritium-containing compounds, including compounds containing radioactive isotopes, including radioactive isotopes effective in therapy and diagnosis; multimers of the compound, including dimers, trimers, etc.; salts of the compound, preferably pharmaceutically acceptable salts, including acid addition salts and base addition salts, including salts with organic and inorganic counterions, including zwitterionic forms, and where the compound is associated with two or more counterions, the two or more counterions may be the same or different; and solvates of the compound, including hemisolvates, monosolvates, disolvates, etc., including organic and inorganic solvates, where the inorganic solvates include hydrates, and where the compound is associated with two or more solvent molecules, the two or more molecules may be the same or different. In some instances, reference herein to a compound of the invention includes reference to one of the above forms or, for example, an explicit reference to a salt and / or solvate. However, this reference is for emphasis only and should not be construed as excluding other of the above-specified forms.

[0069] "Pharmaceutical product" generally, but not necessarily always, means the final formulation containing the active drug ingredient (i.e., the liposome containing the IDO inhibitor prodrug) together with the inactive ingredients. This term also includes the final dosage form that does not contain the active ingredient but is intended to be used as a placebo.

[0070] The term "disulfide" can refer to the -S-S- group.

[0071] The term "empty vesicle" means the unloaded lipid vesicle itself.

[0072] As used herein, the term "ester" means a chemical compound derived from an acid (organic or inorganic) in which at least one -OH hydroxyl group has been replaced by an -O-alkyl (alkoxy) or O-aryl (aryloxy) group.

[0073] As used herein, the term "esterase" is a hydrolase that splits an ester into an acid and an alcohol.

[0074] "Excipient" means an inert substance used as a carrier for the active ingredient in a drug such as a vaccine. Excipients may also be used to bulk up a formulation having a very potent active ingredient in order to allow for convenient and accurate dosing. Examples of excipients include, but are not limited to, anti-adhesion agents, binders, coatings, disintegrants, fillers, diluents, flavoring agents, coloring agents, lubricants and preservatives.

[0075] As used herein, the term "halo", "halide" or "halogen" refers to fluoro, chloro, bromo and iodo groups.

[0076] The terms "hydroxyl" and "hydroxy" refer to the -OH group.

[0077] As used herein, the terms "inhibit" or "inhibition of" mean to reduce by a measurable amount or to completely suppress.

[0078] As used in the context of the present disclosure, the terms "individual" or "patient" can be used interchangeably.

[0079] As used herein, the term "ligand" generally refers to a species, such as a molecule or ion, that interacts (e.g., binds) with another species in some way. See Martell, A.E., and Hancock, R.P., Metal Complexes in Aqueous Solutions, Plenum: New York (1996), which is incorporated herein by reference in its entirety.

[0080] As used herein, the term "lipid" refers to a class of naturally occurring (organic) compounds that are insoluble in polar solvents. In the context of the present disclosure, lipids refer to normal lipids, phospholipids, cholesterol, PEG, and chemically functionalized lipids for attaching ligands, etc.

[0081] The term "lipid bilayer" or "LB" refers to any bilayer of oriented amphiphilic lipid molecules in which the hydrocarbon tails face inward to form a continuous nonpolar phase.

[0082] The terms "liposome" or "lipid vesicle" or "vesicle" are used interchangeably to refer to an aqueous compartment enclosed by a lipid bilayer, as conventionally defined (see Stryer (1981) Biochemistry, 2d Edition, W.H. Freeman & Co., p. 213).

[0083] The term "mammal" refers to any organism classified as a mammal, including mice, rats, rabbits, dogs, cats, cows, horses, and humans. In one embodiment of the invention, the mammal is a mouse. In another embodiment of the invention, the mammal is a human.

[0084] The terms "mercapto" or "thiol" refer to the -SH group.

[0085] The terms "metastatic cancer" and "metastatic disease" mean cancer that has metastasized to regional lymph nodes or distant sites, and shall include diseases of stage D in the AUA system and stage T×N×M+ in the TNM system.

[0086] The terms "nanocarrier", "nanoparticle" and "nanoparticle drug carrier" are used interchangeably and refer to nanostructures having an aqueous, solid or polymeric inner core. In certain embodiments, the nanocarrier comprises a lipid bilayer that encloses (or surrounds or encapsulates) a porous particle core. In certain embodiments, the nanocarrier is a liposome, a lipid nanoparticle ("LNP") or a solid-lipid nanoparticle ("SLNP").

[0087] The terms "nanoscale particle", "nanomaterials", "nanocarrier" and "nanoparticle" refer to a structure having at least one region with dimensions (e.g., length, width, diameter, etc.) of less than about 1,000 nm. In some embodiments, the dimensions are smaller (e.g., less than about 500 nm, less than about 250 nm, less than about 200 nm, less than about 150 nm, less than about 125 nm, less than about 100 nm, less than about 80 nm, less than about 70 nm, less than about 60 nm, less than about 50 nm, less than about 40 nm, less than about 30 nm, or less than about 20 nm). In some embodiments, the dimensions are from about 20 nm to about 250 nm (e.g., about 20, 30, 40, 50, 60, 70, 80, 90, 100, 110, 120, 130, 140, 150, 160, 170, 180, 190, 200, 210, 220, 230, 240 or 250 nm).

[0088] The term "nanovesicle" refers to a "lipid vesicle" having a diameter in the range of from about 20 nm, or from about 30 nm, or from about 40 nm, or from about 50 nm to about 500 nm, or to about 400 nm, or to about 300 nm, or to about 200 nm, or to about 150 nm, or to about 100 nm, or to about 80 nm (or a collection of vesicles having an average diameter). In certain embodiments, the nanovesicles have a diameter in the range of about 40 nm to about 80 nm, or about 50 nm to about 70 nm.

[0089] "Pharmaceutically acceptable" refers to non-toxic, inert, and / or compositions that are physiologically compatible with humans or other mammals.

[0090] "Pharmaceutical formulation" means the process of combining different chemical substances into a pure drug substance to produce a final pharmaceutical product.

[0091] The term "phosphonate" refers to a -P(=O)(OR)2 group, where each R is independently H, alkyl, aralkyl, aryl, or a negative charge (i.e., there is no R group substantially bonded to an oxygen atom, resulting in the presence of an unshared electron pair on the oxygen atom). Thus, in other words, each R can be present or absent, and if present, is selected from H, alkyl, aralkyl, or aryl.

[0092] The term "phosphate" refers to a -OP(=O)(OR’)2 group, where R’ is H or a negative charge.

[0093] The term "prodrug" means a drug agent or compound that is metabolized into a pharmacologically active drug after administration. For the purposes of the present disclosure, the prodrugs of the present invention comprise three components: (i) a drug moiety; (ii) a lipid moiety; and (iii) a linking unit ("LU").

[0094] The term "IDO prodrug" means a prodrug of the present invention in which the drug moiety comprises an IDO inhibitor.

[0095] The term "pyrolipid" refers to a conjugate of a lipid and a porphyrin, a porphyrin derivative or a porphyrin analog. In some embodiments, the pyrolipid can include a lipid conjugate in which a porphyrin or its derivative or analog is covalently bonded to a lipid side chain. See, for example, U.S. Patent Application Publication No. 2014 / 0127763.

[0096] As used herein, the terms "specific", "specifically binds" and "specifically binds to" refer to the selective binding of the nanocarriers of the present invention to the target IDO-1.

[0097] The term "supported lipid bilayer" means a lipid bilayer surrounding a porous particle core. This definition as described in the present disclosure is stated because the lipid bilayer is located on the surface and supported by the porous particle core. In certain embodiments, the lipid bilayer can have a thickness in the range of about 6 nm to about 7 nm, which includes a hydrophobic core having a thickness of 3 - 4 nm, a hydrated hydrophilic headgroup layer (each about 0.9 nm), and two partially hydrated regions each about 0.3 nm. In various embodiments, the lipid bilayer surrounding the liposome includes a continuous bilayer or a substantially continuous bilayer that effectively encapsulates and seals the IDO inhibitor.

[0098] The term "thioalkyl" can refer to the group -SR, where R is selected from H, alkyl, substituted alkyl, aralkyl, substituted aralkyl, aryl and substituted aryl. Similarly, the terms "thioaralkyl" and "thioaryl" refer to the -SR group, where R is aralkyl and aryl, respectively.

[0099] As used herein, the term "treating" or "therapeutic" and grammatically related terms refer to any improvement in any outcome of a disease, such as an extension of survival, a decrease in morbidity, and / or a reduction in side effects that are a byproduct of an alternative treatment modality. As will be readily understood in the art, complete eradication of the disease is preferred but not a requirement of the treatment act.

[0100] The term "therapeutically effective amount" refers to the amount of an active prodrug, nano-encapsulated prodrug or pharmaceutical agent that elicits a biological or medical response in a tissue, system, animal, individual or human.

[0101] The term "unsupported lipid bilayer" means an uncoated lipid bilayer in a lipid vesicle or liposome. II.) Prodrug

[0102] As shown in the present disclosure, and for the purposes of the present invention, suitable prodrugs are formed by conjugating the drug moiety of the present invention (see the section entitled "Drug Moiety") to the lipid moiety of the present invention (see the section entitled "Lipid") via the LU (see the section entitled "Linking Unit") of the present disclosure. For the purposes of the present disclosure, several strategies can be utilized for the formation of IDO prodrugs. (See, for example, FIGS. 4, 5 and 6).

[0103] Thus, in some embodiments, the prodrug is a drug-lipid moiety comprising an IDO inhibitor of the present disclosure.

[0104] In one embodiment, the prodrug comprises the following chemical structure represented by Formula I: [Chemical Formula] Wherein, in an exemplary embodiment of Formula I: X1 = Cl, F, CN; X2 = H, F; and A, B = H, CH3; Thus, in one embodiment, the prodrug is a drug-lipid moiety comprising an IDO inhibitor of Formula I.

[0105] In one embodiment, the prodrug is a drug-lipid moiety comprising an IDO inhibitor described in FIG. 4.

[0106] In one embodiment, the prodrug is a drug-lipid moiety comprising the IDO inhibitor described in FIG. 5.

[0107] In one embodiment, the prodrug is a drug-lipid moiety comprising the IDO inhibitor described in FIG. 6.

[0108] In a further embodiment, the IDO prodrug is a drug-lipid moiety comprising a lipid of the present disclosure.

[0109] In a further embodiment, the IDO prodrug is a drug-lipid moiety wherein the lipid is CHEMS.

[0110] In a further embodiment, the IDO prodrug is a drug-lipid moiety comprising an LU of the present disclosure.

[0111] In a further embodiment, the IDO prodrug is a drug-lipid moiety wherein the LU is a hydroxymethylcarbamate linker.

[0112] In a further embodiment, the prodrug is a drug-lipid moiety comprising an IDO inhibitor of the present invention, wherein the IDO inhibitor comprises chemical composition ID3.

[0113] In a further embodiment, the prodrug is a drug-lipid moiety comprising an IDO inhibitor of the present invention, wherein the IDO inhibitor comprises ID3 and further comprises CHEMS.

[0114] In a further embodiment, the prodrug is a drug-lipid moiety comprising an IDO inhibitor of the present invention, wherein the IDO inhibitor comprises ID3 and further comprises stearic acid.

[0115] In a further embodiment, the prodrug is a drug-lipid moiety comprising an IDO inhibitor of the present invention, wherein the IDO inhibitor comprises ID3, further comprises CHEMS, and the LU is a hydroxymethylcarbamate linker.

[0116] In a further embodiment, the prodrug is a drug-lipid moiety comprising an IDO inhibitor of the present invention, wherein the IDO inhibitor comprises ID3, further comprises stearic acid, and LU is a hydroxymethylcarbamate linker, which is a drug-lipid moiety.

[0117] In a further embodiment, the prodrug is a drug-lipid moiety comprising an IDO inhibitor of the present invention, wherein the IDO inhibitor comprises ID3, further comprises stearic acid, and has the following structure:

Chemical formula

[0118] In a further embodiment, the prodrug is a drug-lipid moiety comprising an IDO inhibitor of the present invention, wherein the IDO inhibitor comprises ID3, further comprises cholesterol hemisuccinate, and has the following structure:

Chemical formula

[0119] In a further embodiment of the present disclosure, the subject provides an IDO inhibitor prodrug comprising a therapeutically active agent prodrug conjugated with a lipid. In some embodiments, the prodrug comprises (a) a monovalent drug moiety, (b) a monovalent lipid moiety, and (c) a divalent linker moiety comprising a linking unit that decomposes in vivo, such as a disulfide bond, wherein the monovalent drug moiety and the monovalent lipid moiety are linked (e.g., covalently bonded) via the linker. The monovalent drug moiety and the monovalent lipid moiety can each be a monovalent derivative of a chemical compound and a lipid, respectively. For example, the monovalent derivative can be a deprotonated derivative of a chemical compound or lipid comprising a hydroxyl, thiol, amino, or carboxylic acid group.

[0120] In a further embodiment of the disclosure, the subject provides an IDO inhibitor prodrug comprising a lipid-conjugated therapeutic agent prodrug. In some embodiments, the prodrug comprises a (a) divalent drug moiety, (b) divalent lipid moiety, and (c) a divalent linker moiety comprising a linkage that degrades in vivo, wherein the divalent drug moiety and the divalent lipid moiety are linked (e.g., covalently bonded) via the linker. The divalent drug moiety and the divalent lipid moiety can each be a divalent derivative of a chemical compound and a lipid, respectively. For example, the divalent derivative can be a deprotonated derivative of a chemical compound or lipid that contains a hydroxyl, thiol, amino, or carboxylic acid group.

[0121] Those skilled in the art will understand and be enabled to make variations and modifications to the disclosed embodiments without departing from the functions and objectives of the invention disclosed herein. Such variations and modifications are intended to be within the scope of the present disclosure. III.) Drug Moiety

[0122] Another aspect of the invention provides a novel IDO prodrug compound having the following formula represented by ID3.

[0123] Those skilled in the art will understand that the compound is useful as an IDO inhibitor (e.g., inhibits IDO-1). As a brief background, IDO-1 is involved in immune regulation through its ability to limit T cell function and function in the mechanism of immune tolerance. Munn et al., Trends in See Immunology, 34(3) pp. 137-143 (2012). New evidence suggests that IDO is activated during tumorigenesis and helps malignant cells escape eradication by the immune system. In mice, IDO has a normal immune checkpoint function in pregnancy-induced immune tolerance and suppresses the mother's immune system. See YU et al., Cellular Physiology and Biochemistry, 49(1): pp. 134-143 (2018). Studies have shown that overexpression of IDO in several tumors such as ovarian cancer, colorectal cancer, endometrial cancer, and esophageal cancer correlates with earlier death, while in kidney cancer and liver cancer, overexpression of IDO appears to correlate with better outcomes. Ibid.

[0124] Based on the above, the present disclosure describes a class of IDO inhibitors.

[0125] In one embodiment, the drug moiety of the present disclosure comprises a compound having the following chemical structure (represented by ID3):

Chemical formula

[0126] Those skilled in the art will understand and be able to make variations and modifications to the disclosed embodiments without changing the functions and objectives of the invention disclosed herein. Such variations and modifications are intended to be within the scope of the present disclosure. IV.) Lipids

[0127] Generally speaking, and for the purposes of the present disclosure, the term "lipid" is used in its broadest sense and includes several subcategories of lipids including, but not limited to, phospholipids / fatty acids. As will be understood by those skilled in the art, phospholipids correspond to a class of lipids that are a major component of all cell membranes. Phospholipids can form lipid bilayers due to their amphiphilic properties. The structure of a phospholipid molecule generally consists of two hydrophobic fatty acid "tails" and a hydrophilic "head" consisting of a phosphate group that can be modified by simple organic molecules such as choline, ethanolamine or serine. These two components are usually linked together by a glycerol molecule. A representative list of the phospholipids / fatty acids of the present invention is shown in Table III.

[0128] As a simple background, at the most fundamental level, the properties of liposomes depend on the delicate physicochemical interactions between the various lipid species in their composition. Individual lipids can combine to form countless higher-order structures including bilayers, and the properties of the bilayers can be adjusted to regulate drug release and membrane stability. In a simplified bilayer model, the acyl chain length defines the bilayer thickness and the phase transition temperature (Tm), the saturation of the acyl chains controls the bilayer fluidity, and the head group interactions affect the intermolecular and intramolecular forces between the lipids. Liposome behavior can be adjusted by incorporating synthetic lipids such as lipid prodrugs, membrane-fusogenic lipids and functionalizable lipids into the bilayer. See Kohli et al., J. Control Release, 0: pp. 274-287 (Sept. 28, 2014).

[0129] In one embodiment of the present disclosure, the IDO prodrug comprises a monovalent lipid moiety.

[0130] In one embodiment, the IDO prodrug comprises a divalent lipid moiety.

[0131] In one embodiment, the lipid comprises cholesterol having the following chemical structure.

Chemical formula

[0132] In one embodiment, the lipid includes DPPG having the following chemical structure.

Chem.

[0133] In one embodiment, the lipid includes DMPG having the following chemical structure.

Chem.

[0134] In one embodiment, the lipid includes lyso-PC having the following chemical structure.

Chem.

[0135] In one embodiment, the lipid includes (Δ9-cis)PG having the following chemical structure.

Chem.

[0136] In one embodiment, the lipid includes soy lyso-PC having the following chemical structure.

Chem.

[0137] In one embodiment, the lipid includes PG having the following chemical structure.

Chem.

[0138] In one embodiment, the lipid includes C16 PEG2000 ceramide having the following chemical structure.

Chem.

[0139] In one embodiment, the lipid comprises cholesterol hemisuccinate ("CHEMS") having the following chemical structure.

Chemical formula

[0140] In one embodiment, the lipid comprises stearic acid having the following chemical structure.

Chemical formula

[0141] For reference, a complete list of the chemical formulas and abbreviations of the lipids disclosed herein is shown in Table I.

[0142] In a further embodiment, the lipid comprises a phospholipid / fatty acid disclosed herein and shown in Table III.

[0143] Furthermore, the IDO prodrugs and / or liposomes of the present disclosure may contain one or more helper lipids, also referred to herein as "helper lipid components". The helper lipid component is preferably selected from the group comprising phospholipids and steroids. The phospholipids are preferably diesters and monoesters of phosphoric acid. Preferred members of the phospholipids are phosphoglycerides and sphingolipids. Steroids, as used herein, are naturally occurring and synthetic compounds based on partially hydrogenated cyclopenta[a]phenanthrene. Preferably, the steroid contains 21 to 30 carbon atoms. A particularly preferred steroid is cholesterol.

[0144] While not wishing to be bound by any theory, it is noted that due to the specific molar percentage of helper lipids contained in the lipid composition according to the present invention, this helper lipid can be either a PEG-free helper lipid or particularly a PEG-containing helper lipid, and more specifically, surprising effects can be achieved if the content of any of this type of helper lipid is within the concentration range specified herein.

[0145] In a further aspect of the present invention, the lipid composition, preferably present as a lipoplex or liposome, preferably exhibits a neutral or overall anionic charge. The anionic lipid is preferably any neutral or anionic lipid described herein. The lipid composition, in a preferred embodiment, contains any IDO inhibitor described herein in addition to any helper lipid or combination of helper lipids. In a further embodiment, the composition according to the present invention containing a nucleic acid forms a lipoplex. In a preferred embodiment, the term lipoplex as used herein refers to a composition composed of the neutral or anionic lipid, neutral helper lipid and IDO inhibitor of the present invention. For reference regarding the use of helper lipids in the art, see, for example, US Patent Application Publication No. 2011 / 0178164; Ojeda et al., Int. J. of Pharmaceutics (March 2016); Dabkowska et al., J. R. Soc. Interface 9, pp. 548 - 561 (2012); and Mochizuki et al., Biochimica et. Biophysica Acta, 1828, pp. 412 - 418 (2013).

[0146] In a preferred embodiment, the helper lipid of the present invention includes the helper lipids described in Table II.

[0147] In one embodiment, the IDO prodrug contains the lipid of the present invention, the lipid is CHEMS, and the drug moiety is ID3.

[0148] In one embodiment, the IDO prodrug comprises a lipid of the present invention, the lipid is CHEMS, the drug moiety is ID3, further comprises LU, and LU is a hydroxymethylcarbamate linker.

[0149] In one embodiment, the IDO prodrug comprises a lipid of the present invention, the lipid is CHEMS, the drug moiety is ID3, further comprises LU, LU is a hydroxymethylcarbamate linker, and further comprises a helper lipid component, and the helper lipid component comprises the helper lipids in Table II.

[0150] In one embodiment, the IDO prodrug comprises a lipid of the present invention, the lipid is CHEMS, the drug moiety is ID3, and CHEMS is monovalent.

[0151] In one embodiment, the IDO prodrug comprises a lipid of the present invention, the lipid is stearic acid, and the drug moiety is ID3.

[0152] In one embodiment, the IDO prodrug comprises a lipid of the present invention, the lipid is stearic acid, the drug moiety is ID3, and stearic acid is monovalent.

[0153] In one embodiment, the IDO prodrug comprises a lipid of the present invention, the lipid is stearic acid, the drug moiety is ID3, further comprises LU, and LU is a hydroxymethylcarbamate linker.

[0154] In one embodiment, the IDO prodrug comprises a lipid of the present invention, the lipid is stearic acid, the chemical composition is ID3, further comprises LU, LU is a hydroxymethylcarbamate linker, and further comprises a helper lipid component, and the helper lipid component comprises the helper lipids in Table II.

[0155] Those skilled in the art will understand and be able to make modifications and variations to the disclosed embodiments without changing the functions and purposes of the present invention disclosed herein. Such modifications and variations are intended to be within the scope of this disclosure. V.) Linking Unit (the "LU")

[0156] In some embodiments, the subject matter of the present disclosure provides prodrugs comprising drug-lipid conjugates that include biodegradable linkages such as esters, thioesters, and other linkers known in the art.

[0157] Exemplary embodiments of ester chemistry are described herein. [Chemical formula]

[0158] In some embodiments, the prodrug is a drug-lipid conjugate, and the drug-lipid conjugate is cleaved by an esterase.

[0159] In one embodiment, the prodrug of the present invention comprises an LU via a secondary amine, amide, or aniline using the following scheme. [Chemical formula] Exemplary syntheses are as follows. [Chemical formula] Cleavage of the prodrug structure containing a secondary amine, amide, or aniline is obtained by esterase hydrolysis of the secondary amine, amide, or aniline prodrug under the following exemplary syntheses. [Chemical formula] wherein R1 and R2 can be molecules that connect N via C.

[0160] In one embodiment, the secondary amide nitrogen of the ID3 drug moiety is conjugated to CHEMS via a hydroxymethylcarbamate linker.

[0161] In one embodiment, the secondary amide nitrogen of the ID3 drug moiety is conjugated to stearic acid via a hydroxymethylcarbamate linker.

[0162] Those skilled in the art will understand and be enabled to make variations and modifications to the disclosed embodiments without departing from the functions and objectives of the invention disclosed herein. Such variations and modifications are intended to be within the scope of this disclosure. VI.) Nanocarriers

[0163] Generally speaking, and for the purposes of this disclosure, nanocarriers are within the scope of the present invention. Nanocarriers are nanomaterials that are used as transport modules for another substance such as a drug. Commonly used nanocarriers include micelles, polymers, carbon-based materials, liposomes and other substances. Due to their small size, nanocarriers can deliver drugs to sites in the body that would otherwise be inaccessible. Nanocarriers can include polymer conjugates, polymer nanoparticles, lipid-based carriers, dendrimers, carbon nanotubes and gold nanoparticles. Lipid-based carriers include both liposomes and micelles.

[0164] Furthermore, nanocarriers can be useful in the drug delivery process because they can deliver drugs to site-specific targets, delivering the drug to a particular organ or cell and not to other organs or cells. Site specificity is a major therapeutic advantage as it prevents the drug from being delivered to the wrong place. Additionally, nanocarriers can be promising for use in chemotherapy as they can help reduce the harmful and more widespread toxicity of chemotherapy to rapidly growing healthy cells in the body. Since chemotherapeutic drugs can be extremely toxic to human cells, it is important that the chemotherapeutic drug be delivered to the tumor without being released into other parts of the body.

[0165] Generally speaking, there are four ways in which nanocarriers can deliver drugs, including passive targeting, active targeting, pH specificity, and temperature specificity.

[0166] Passive targeting refers to the ability of nanocarriers to enter the tumor vasculature, be captured, and accumulate in the tumor. This accumulation is caused by the enhanced permeability and retention effect. The leaky vasculature of tumors is a network of blood vessels formed within the tumor and contains many small pores. These pores can allow nanocarriers to enter, but also contain many bends that enable the nanocarriers to be captured. As more nanocarriers are captured, drugs accumulate at the tumor site. This accumulation results in a large amount of drugs being directly delivered to the tumor site.

[0167] Active targeting involves incorporating targeting modules, such as ligands or antibodies, that are specific for certain types of cells in the body, onto the surface of the nanocarriers. Generally, nanocarriers have a high surface area to volume ratio, allowing multiple ligands to be incorporated onto their surface.

[0168] Furthermore, certain nanocarriers release the drugs they contain only within a specific pH range. The pH specificity also enables the nanocarriers to deliver drugs directly to the tumor site. This is due to the fact that tumors are generally more acidic than normal human cells, with a pH of about 6.8, while normal tissues have a pH of about 7.4. Therefore, nanocarriers that release drugs only within a specific pH range can be used to release drugs only within the acidic tumor environment. The highly acidic environment causes the release of drugs by degrading the structure of the nanocarriers. Generally, these nanocarriers do not release drugs in a neutral or basic environment, effectively targeting the acidic environment of the tumor and leaving normal body cells intact. This pH sensitivity can also be induced in micelle systems by adding copolymer chains to micelles that have been found to act in a pH-independent manner. See WU et al., Biomaterials, 34(4):1213-1222 (2012). These micelle-polymer conjugates also help prevent cancer cells from developing multi-drug resistance. The low pH environment triggers the rapid release of the micelle polymer, releasing most of the drug at once rather than gradually as in other drug treatments.

[0169] Furthermore, some nanocarriers have also been shown to deliver drugs more effectively at a specific temperature. Since tumor temperature is generally about 40°C, which is higher than the temperature of the rest of the body, this temperature gradient serves to function as a safety device for tumor-specific site delivery. See REZAEI et al., Polymer, 53(16):3485-3497 (2012).

[0170] As disclosed herein, lipid-based nanocarriers such as liposomes are within the scope of the present invention. Lipid-based nanoparticles (LBNP or LNP) such as liposomes, solid lipid nanoparticles (SLN) and nanostructured lipid carriers (NLC) transport hydrophobic and hydrophilic molecules, exhibit very low toxicity or no toxicity, and can increase the duration of drug action by prolonging the half-life and controlled release of drugs. Lipid nanoparticles can include chemical modifications for avoiding detection by the immune system (such as gangliosides or polyethylene glycol (PEG)) or for improving the solubility of drugs. Furthermore, lipid nanoparticles can be prepared in pH-sensitive formulations to promote drug release in acidic environments and can also associate with small molecules or antibodies (such as folic acid (FoA)) that recognize tumor cells or their receptors. To improve the patient response, nanodrugs can also be used in combination with other therapeutic strategies. GARCIA-PINEL et al., Nanomaterials See 9(639)(2019).

[0171] In various embodiments, the silica-some drug carriers described herein include porous silica (or other material) nanoparticles coated with a lipid bilayer (e.g., a silica body having a surface and defining a plurality of pores suitable for receiving molecules therein). The fact that the nanoparticles are referred to as silica nanoparticles does not exclude the incorporation of materials other than silica within the silica nanoparticles. In some embodiments, the silica nanoparticles can be substantially spherical with a plurality of pore openings leading to a surface that provides access to the pores. However, in various embodiments, the silica nanoparticles can have a shape other than substantially spherical. Thus, for example, in certain embodiments, the silica nanoparticles can be substantially oval, rod-shaped, substantially regular polygon, irregular polygon, etc.

[0172] Generally, silica nanoparticles include a silica body that defines an outer surface between pore openings and sidewalls within the pores. The pores can extend through the silica body to another pore opening, or the pores can extend only partially through the silica body such that the pores have a bottom surface defined by the silica body.

[0173] In some embodiments, the silica body is mesoporous. In other embodiments, the silica body is microporous. As used herein, "mesoporous" means having pores with diameters from about 2 nm to about 50 nm, and "microporous" means having pores with diameters less than about 2 nm. Generally, the pores can be of any size, but in typical embodiments, they are large enough to contain one or more therapeutic compounds therein. In such embodiments, the pores allow small molecules, such as therapeutic compounds like anti-cancer compounds, to adhere or bind to the inner surface of the pores and be released from the silica body when used for therapeutic purposes. In some embodiments, the pores are substantially cylindrical.

[0174] In certain embodiments, the nanoparticles include pores having a pore diameter of about 1 nm to about 10 nm or about 2 nm to about 8 nm. In certain embodiments, the nanoparticles include pores having a pore diameter of about 1 nm to about 6 nm or about 2 nm to about 5 nm. Other embodiments include particles having a pore diameter of less than 2.5 nm.

[0175] In other embodiments, the pore diameter is 1.5 to 2.5 nm. Silica nanoparticles having other pore sizes can be prepared, for example, by using different surfactants or swelling agents during the preparation of the silica nanoparticles. In various embodiments, the nanoparticles can have a size of about 1000 nm (e.g., including particles with an average or median diameter (or another characteristic dimension). However, in various embodiments, generally particles larger than 300 nm may not be very effective when entering living cells or fenestrations of blood vessels, so the nanoparticles are typically less than 500 nm or about less than 300 nm. In certain embodiments, the nanoparticles range in size from about 40 nm, or about 50 nm, or about 60 nm to about 100 nm, or up to about 90 nm, or up to about 80 nm, or up to about 70 nm. In certain embodiments, the nanoparticles range in size from about 60 nm to about 70 nm. Some embodiments include nanoparticles having an average maximum dimension from about 50 nm to about 1000 nm. Other embodiments include nanoparticles having an average maximum dimension from about 50 nm to about 500 nm. Other embodiments include nanoparticles having an average maximum dimension from about 50 nm to about 200 nm.

[0176] In some embodiments, the average maximum dimension is greater than about 20 nm, greater than about 30 nm, greater than 40 nm, or greater than about 50 nm. Other embodiments include nanoparticles having an average maximum dimension less than about 500 nm, less than about 300 nm, less than about 200 nm, less than about 100 nm or less than about 75 nm. As used herein, the size of the nanoparticles refers to the average or median size of the primary particles measured by transmission electron microscopy (TEM) or similar visualization techniques known in the art. Further examples of mesoporous silica nanoparticles include, but are not limited to, MCM-41, MCM-48, and SBA-15. See KATIYARE et al., J. Chromotog. 1122(1-2):13-20(2006).

[0177] Methods for producing porous silica nanoparticles are well known to those skilled in the art. In certain embodiments, mesoporous silica nanoparticles are synthesized by reacting tetraethyl orthosilicate (TEOS) with a template made of micellar rods. As a result, an assembly of nano-sized spheres or rods filled with a regular arrangement of pores is obtained. The template can then be removed by washing with a solvent adjusted to an appropriate pH (see, for example, TREWYN et al., (2007) Chem. Eng. J. 137(1):23-29).

[0178] In certain embodiments, mesoporous particles can also be synthesized using a simple sol-gel method (see, for example, NANDIYANTO et al., (2009) Microporous and Mesoporous Mat. 120(3):447-453). In certain embodiments, tetraethyl orthosilicate can also be used with an additional polymer monomer as a template. In certain embodiments, 3-mercaptopropyl)trimethoxysilane (MPTMS) is used instead of TEOS.

[0179] In certain embodiments, the mesoporous silica nanoparticles are the core and are synthesized by a modification of the sol / gel procedure described by MENG et al., (2015) ACS Nemo, 9(4):3540-3557.

[0180] The methods described herein have been demonstrated for porous silica nanoparticles (e.g., mesoporous silica), but it will be recognized by those skilled in the art that similar methods can be used with other porous nanoparticles. A number of other mesoporous materials that can be used in drug delivery nanoparticles are known to those skilled in the art. For example, in certain embodiments, mesoporous carbon nanoparticles can be utilized.

[0181] Mesoporous carbon nanoparticles are well known to those skilled in the art (see, for example, HUANG et al., (2016) Carbon, 101:135-142; ZHU et al., (2014) Asian J. Pharm. Sci., 9(2):82-91, etc.).

[0182] Similarly, in certain embodiments, mesoporous polymer particles can be utilized. The synthesis of highly ordered mesoporous polymers and carbon frameworks from the organic-organic organization of triblock copolymers with soluble low molecular weight phenolic resin precursors (resols) by an evaporation-induced self-assembly strategy has been reported by MENG et al., (2006) Chem. Mat. 6(18):4447-4464.

[0183] The nanoparticles described herein are exemplary and non-limiting. Using the teachings provided herein, numerous other lipid bilayer-coated nanoparticles are available to those skilled in the art.

[0184] In one embodiment, the present invention teaches a nanocarrier comprising an IDO prodrug.

[0185] In one embodiment, the present invention teaches a nanocarrier comprising liposomes, wherein the lipid comprises CHEMS.

[0186] In one embodiment, the present invention teaches a nanocarrier comprising liposomes, wherein the lipid comprises stearic acid.

[0187] In one embodiment, the present invention teaches a nanocarrier comprising liposomes, wherein the lipid comprises CHEMS and the liposomes further comprise an IDO prodrug.

[0188] In one embodiment, the present invention teaches a nanocarrier comprising liposomes, wherein the lipid comprises CHEMS and the liposomes further comprise ID3.

[0189] In one embodiment, the present invention teaches a nanocarrier comprising liposomes, wherein the lipid comprises stearic acid and the liposomes further comprise an IDO inhibitor.

[0190] In one embodiment, the present invention teaches a nanocarrier comprising liposomes, wherein the lipid comprises stearic acid and the liposomes further comprise ID3.

[0191] The scope of the present disclosure teaches three possible treatment modalities using the formulated prodrugs of the present invention. See International Publication No. WO 2018 / 213631.

[0192] The first treatment modality involves a combination of an IDO prodrug combined with another therapeutic agent (e.g., another formulated prodrug that inhibits IDO-1, a chemotherapeutic agent such as ICD-inducing chemotherapy, etc.) in a single liposome that enables systemic (or local) biodistribution and drug delivery to the tumor site. The dual delivery approach achieved a synergistic enhancement of adaptive and innate immunity, resulting in a significant improvement in animal survival. In certain embodiments, the nanocarrier comprises vesicles (i.e., lipid bilayers encapsulating fluid).

[0193] The second treatment modality involves local delivery of an agent that inhibits IDO-1, combined with a lipid (e.g., liposome) that contains an inhibitor of IDO-1, to the tumor or the peritumoral region. Such local delivery of an IDO-1 inhibitor in combination with an IDO prodrug has been demonstrated to induce cytotoxic tumor killing and tumor shrinkage at the local site. These adaptive immune responses are accompanied by activation of a DC population that is particularly well-suited for generating cytotoxic T cell responses, in addition to enhancement of the innate immune system as reflected by CRT expression.

[0194] The third treatment modality involves vaccination using dying cancer cells (e.g., KPC cells) in which IDO-1 inhibition is induced ex vivo. Such vaccination has been found to be able to interfere with tumor growth at distant sites and to generate a systemic immune response that can enable adoptive transfer into non-immunized animals. Those skilled in the art will understand and be enabled to implement the method treatment modalities provided herein. VII.) Liposomes

[0195] In one aspect, the subject matter of the present disclosure is based on an approach for providing prodrugs of the present disclosure (see the section entitled prodrugs) suitable for incorporation into nanocarriers comprising a lipid coating layer to enhance the delivery of the corresponding prodrugs and to provide combination therapies comprising the prodrugs. Advantages of using the prodrugs of the present invention include facilitating controlled formulation into the LNPs (e.g., liposomes) of the present disclosure. Thereby, the prodrug can be kept in an inactive form in the systemic circulation, whereby the liposome can release the active agent, for example, after phagocytosis by cells within the tumor.

[0196] In certain embodiments, one or more IDO prodrugs (e.g., any one or more of the IDO prodrug inhibitors taught by Formula I and / or ID3) (see the section entitled prodrugs) are formulated with a lipid moiety capable of forming a vesicle (e.g., liposome) structure in an aqueous solution or forming a component of the lipid bilayer that constitutes the liposome. The liposomes can be used directly and provided as a component in a combined formulation (e.g., in combination with another drug moiety or treatment modality disclosed herein).

[0197] In certain embodiments, the liposomes formulated with the IDO prodrug comprise lipids, PHGP, vitamin E, cholesterol, and / or fatty acids.

[0198] In one embodiment, the liposome contains cholesterol.

[0199] In one embodiment, the liposome contains DPPG.

[0200] In one embodiment, the liposome contains DMPG.

[0201] In one embodiment, the liposome is lysophosphatidylcholine (lyso-PC).

[0202] In one embodiment, the liposome is (Δ9-cis) phosphatidylglycerol ((Δ9-cis) PG).

[0203] In one embodiment, the liposome contains soy lysophosphatidylcholine.

[0204] In one embodiment, the liposome contains phosphatidylglycerol (PG).

[0205] In one embodiment, the liposome contains PA-PEG3-mannose.

[0206] In one embodiment, the liposome contains C16 PEG2000 ceramide.

[0207] In one embodiment, the liposome contains MPLA.

[0208] In one embodiment, the liposome contains CHEMS.

[0209] In one embodiment, the liposome contains stearic acid.

[0210] In one embodiment, the liposome contains the phospholipids shown in Table III.

[0211] In one embodiment, the liposome contains ID3, further contains CHEMS, and further contains LU, and the LU is a hydroxymethylcarbamate linker.

[0212] In one embodiment, the liposome comprises ID3, further comprises stearic acid, further comprises LU, and the LU is a hydroxymethylcarbamate linker.

[0213] In one embodiment, the liposome comprises ID3, further comprises CHEMS, further comprises LU, the LU is a hydroxymethylcarbamate linker, and further comprises helper lipids shown in Table II.

[0214] In one embodiment, the liposome comprises ID3, further comprises stearic acid, further comprises LU, the LU is a hydroxymethylcarbamate linker, and further comprises helper lipids shown in Table II.

[0215] In one embodiment, the liposomes of the present disclosure comprise an IDO prodrug co-formulated with one or more additional immunomodulatory agents, and the immunomodulatory agents include, but are not limited to, immunogenic cell death-inducing chemotherapeutic agents, toll-like receptor agonists, sting agonists, CTLA4 inhibitors, PD-1 inhibitors, and / or prodrugs thereof.

[0216] In a preferred embodiment, the liposome comprises an IDO prodrug co-formulated with an ICD-inducing chemotherapeutic agent.

[0217] In a preferred embodiment, the liposome comprises an IDO prodrug co-formulated with an ICD-inducing chemotherapeutic agent selected from the list of doxorubicin (DOX), mitoxantrone (MTO), oxaliplatin (OXA), cyclophosphamide (CP), bortezomib, carfilzomib, or paclitaxel.

[0218] In a preferred embodiment, the liposome comprises an IDO prodrug co-formulated with a Toll-like receptor TLR agonist / prodrug.

[0219] In a preferred embodiment, the liposome comprises an IDO prodrug co-formulated with a Toll-like receptor (TLR) agonist / prodrug selected from the following list: Resiquimod (R848), Gardiquimod, 852A, DSR6434, Telratolimod, CU-T12-9, Monophosphoryl lipid A (MPLA), 3D(6-acyl)-PHAD®, SMU127, Pam3CSK4 or 3D-PHAD®.

[0220] In a preferred embodiment, the liposome comprises an IDO prodrug co-formulated with a PD-1 inhibitor / prodrug.

[0221] In a preferred embodiment, the liposome comprises an IDO prodrug co-formulated with a PD-1 inhibitor / prodrug selected from the list of AUNP12, CA-170 or BMS-986189 or prodrugs thereof.

[0222] In a preferred embodiment, the liposome comprises an IDO prodrug co-formulated with doxorubicin (DOX).

[0223] In a preferred embodiment, the liposome comprises an IDO prodrug co-formulated with mitoxantrone (MTO).

[0224] In a preferred embodiment, the liposome comprises an IDO prodrug co-formulated with doxorubicin (DOX) and a PD-1 prodrug.

[0225] In a preferred embodiment, the liposome comprises an IDO prodrug co-formulated with mitoxantrone (MTO) and a PD-1 prodrug.

[0226] In a preferred embodiment, the liposome comprises an IDO prodrug co-formulated with doxorubicin (DOX) and a TLR agonist / prodrug.

[0227] In a preferred embodiment, the liposome comprises an IDO prodrug co-formulated with mitoxantrone (MTO) and a TLR agonist / prodrug.

[0228] In a preferred embodiment, the liposome comprises an IDO prodrug co-formulated with doxorubicin (DOX), a PD-1 prodrug, and a TLR agonist / prodrug.

[0229] In a preferred embodiment, the liposome comprises an IDO prodrug co-formulated with mitoxantrone (MTO), a PD-1 prodrug, and a TLR agonist / prodrug.

[0230] In a preferred embodiment, the liposome comprises an IDO prodrug co-formulated with a TLR agonist / prodrug.

[0231] In a preferred embodiment, the liposome comprises an IDO prodrug co-formulated with a TLR agonist / prodrug.

[0232] In a preferred embodiment, the liposome comprises an IDO prodrug co-formulated with a TLR agonist / prodrug and a PD-1 prodrug.

[0233] In a preferred embodiment, the liposome comprises an IDO prodrug co-formulated with a TLR agonist / prodrug and a PD-1 prodrug.

[0234] In a preferred embodiment, the liposome comprises ID3 co-formulated with doxorubicin (DOX).

[0235] In a preferred embodiment, the liposome comprises ID3 co-formulated with mitoxantrone (MTO).

[0236] In a preferred embodiment, the liposome comprises ID3 co-formulated with doxorubicin (DOX) and / or and IDO prodrug and / or TLR agonist / prodrug.

[0237] In a preferred embodiment, the liposome comprises ID3 co-formulated with mitoxantrone (MTO) and / or and IDO prodrug and / or TLR agonist / prodrug.

[0238] In a preferred embodiment, the liposome comprises ID3 co-formulated with AR5.

[0239] In a preferred embodiment, the liposome comprises ID3 co-formulated with AR5 and TR5.

[0240] In a preferred embodiment, the liposome comprises ID3 co-formulated with NK1.

[0241] In a preferred embodiment, the liposome comprises ID3 co-formulated with NK1 and MTO.

[0242] In a preferred embodiment, the liposome comprises ID3 co-formulated with TR3.

[0243] In a preferred embodiment, the liposome comprises ID3 co-formulated with TR5.

[0244] In a preferred embodiment, the liposome comprises ID3 co-formulated with TB4.

[0245] In a preferred embodiment, the liposome comprises ID3 co-formulated with PD3.

[0246] In a preferred embodiment, the liposome comprises ID3 co-formulated with AR5 and TR3.

[0247] In a preferred embodiment, the liposome comprises ID3 co-formulated with AR5 and TB4.

[0248] In a preferred embodiment, the liposome comprises ID3 co-formulated with AR5 and PD3.

[0249] In another preferred embodiment, the liposome comprises solid-lipid nanoparticles (SLNPs) comprising liposomes containing an IDO prodrug.

[0250] One of ordinary skill in the art recognizes and understands that solubility is one of the most common problems faced by those of ordinary skill in the drug development process. Chemical conjugation of drugs / anticancer agents via lipid molecules (i.e., lipid-based prodrugs) provides a basis for solving the problem of formulating drugs in aqueous suspensions. The main advantage of using lipid conjugation (lipid-based prodrugs) to deliver drugs lies in its ability to improve pharmacokinetics / half-life and targeted delivery.

[0251] By appropriate selection of lipid molecules, lipid-based prodrugs can be incorporated / formulated into liposome formulations using techniques known in the art, which have many additional advantages over conventional drug delivery systems. (KOHLI et al., J. Control Release, 0: pp274-287 (Sept. 28, 2014); and GARCIA-PINEL et al., Nanomaterials 9:638 (2019). The advantages of combining lipid-prodrugs with liposomes consist of two elements: (i) liposomes containing lipid-prodrugs not only increase the solubility of the drug / prodrug itself, but (ii) also have the ability to encapsulate multiple drugs (both hydrophilic and lipophilic) (see the section entitled Nanocarriers).

[0252] For the purposes of the present disclosure, the main advantages of liposome formulations are as follows. i) Biocompatibility / biodegradability and no general toxicity of liposome formulations; (ii) Size, flexibility of surface charge, and manipulation according to the required purpose. The liposome formulation can have a size range of 40 to 150 nm and a surface charge in the range of -40 to +40 mV for the purposes of the present disclosure; and (iii) The liposomes of the present invention have either a single or multiple lipid-prodrugs as the constituent lipid part of the liposome. Furthermore, multiple drugs (e.g., drugs that function by different mechanisms of action) and drugs having different solubility profiles (hydrophilic or lipophilic) can be formulated into these liposomes (either in the lipid bilayer or the hydrophilic core).

[0253] As will be understood by those skilled in the art, all methods of making liposomes involve four basic steps: (i) Drying the lipid from an organic solvent; (ii) Dispersing the lipid in an aqueous solution; (iii) Purifying the resulting liposomes; and (iv) Analyzing the final product. See AKBARZADEH et al., Nanoscale Research Letters, 8:102 (2013).

[0254] Another aspect of the present invention discloses a liposome encapsulation technology (LET), a delivery technology used to deliver drugs. LET is a method of generating ultramicroscopic bubbles called liposomes that encapsulate a number of materials. These "liposomes" form a barrier around the contents of the liposome that is resistant to enzymes, alkaline solutions, digestive juices, bile salts, the intestinal flora produced in the human body, and free radicals in the mouth and stomach. Thus, the contents of the liposome are protected from oxidation and degradation. This protective phospholipid shield or barrier remains intact until the contents of the liposome are delivered to the exact target gland, organ, or system where the contents are to be used (see the section entitled Nanocarriers).

[0255] In one embodiment, the liposomes of the present disclosure are synthesized using multiple different ratios of IDO prodrugs, lipids, and / or lipid prodrugs. As disclosed herein, the IDO prodrug may include the helper lipids disclosed herein (see, e.g., Table II).

[0256] In one embodiment, the liposomes of the present disclosure are synthesized using multiple different ratios of IDO prodrugs, lipids, and / or lipid prodrugs. As disclosed herein, the IDO prodrug may further include DSPE-PEG.

[0257] In a preferred embodiment, the liposomes of the present invention comprise a composition having the following ratios: [Table A]

[0258] In a further preferred embodiment, the liposomes of the present invention comprise a composition having the following ratios: [Table B]

[0259] In a further preferred embodiment, the liposomes of the present invention comprise a composition having the following ratios: [Table C] Here, Lipid 1 includes ID3 and CHEMS.

[0260] In a further preferred embodiment, the liposomes of the present invention comprise a composition having the following ratios: [Table D] Here, Lipid 1 includes ID3 and stearic acid.

[0261] Those skilled in the art will understand and be enabled to make variations and modifications to the disclosed embodiments without departing from the functions and objectives of the invention disclosed herein. Such variations and modifications are intended to be within the scope of this disclosure. VIII.) Pharmaceutical Formulations

[0262] As used herein, the term "drug" is synonymous with "medicine". In certain embodiments, the liposomes of the present disclosure are formulated into an encapsulated dosage form and administered to a patient for the treatment of a disease.

[0263] Generally speaking, "pharmaceutical formulation" is the process of combining different chemical substances into a pure active pharmaceutical ingredient to produce a final pharmaceutical product. Formulation research involves developing a preparation of the drug that is stable and acceptable to the patient. In the case of a drug taken orally, this usually involves incorporating the drug into a tablet or capsule. It is important to understand that the dosage form contains various other substances different from the drug itself, and research must be conducted to ensure the compatibility of the drug with these other substances.

[0264] An excipient is an inert substance used as a carrier for the active ingredient of a pharmaceutical product, in this case, the liposomes containing the IDO prodrug. Additionally, excipients can be used to assist in the manufacturing process of the pharmaceutical product. The active substance is then dissolved or mixed with the excipient. Excipients may also be used to bulk up a formulation with a very potent active ingredient in order to allow for a convenient, accurate dosage. Once the active ingredient is purified, it cannot remain in its purified form for very long. In many cases, the active ingredient denatures, separates from the solution, or adheres to the sides of the container.

[0265] To stabilize the active ingredient, excipients are added to ensure that the active ingredient retains its activity and that the product has a long enough shelf life to be superior to other products and safe for the end user. Examples of excipients include, but are not limited to, anti-adhesion agents, binders, coatings, disintegrants, fillers, diluents, flavoring agents, coloring agents, lubricants, and preservatives. The final formulation contains the active ingredient and excipients, which are then incorporated into a pharmaceutical dosage form.

[0266] Prior to formulation, the physical, chemical, and mechanical properties of the drug are characterized to select what other components should be used in the preparation. Factors such as stability, particle size, polymorphism, pH, and solubility can all affect bioavailability and thus the activity of the drug, and thus formulation studies then consider such factors. The drug must be combined with inert additives in a way that ensures that the amount of drug present is consistent in each dosage unit (e.g., each vial). The dosage should have a uniform appearance.

[0267] It is unlikely that these studies will be completed by the start of clinical trials. This means that first simple preparations are developed for use in Phase I clinical trials. These typically consist of hand-filled capsules containing vials, a small amount of drug, and a diluent. Since these formulations are used (tested) within a few days, proof of their long-term stability is not required. However, since the time the final formulation is packaged until it reaches the patient can be months or years, long-term stability is important in supply chain management. The so-called drug load (i.e., the ratio of the active drug to the total content of the dose) must be considered. A low drug load can cause homogeneity problems. A high drug load can cause flow problems if the compound has a low bulk density or may require large capsules. By the time Phase III clinical trials are reached, the drug formulation should be developed to be close to the preparation that will ultimately be used on the market.

[0268] By this stage, knowledge of stability is essential and conditions must be developed to ensure that the drug is stable in the preparation. Since it is impossible to know what the actual administered dose was, if the drug is found to be unstable, the results obtained from clinical trials will be invalidated. Stability studies are carried out to test whether temperature, humidity, oxidation or photodegradation (ultraviolet or visible light) have any effect, and the preparation is analysed to examine whether degradation products have formed. It is also important to check whether there are any undesirable interactions between the preparation and the container. When using plastic containers, tests are carried out to examine whether any of the components are adsorbed onto the plastic and whether any of the plasticisers, lubricants, pigments or stabilisers leach from the plastic into the preparation. Even the adhesives for the container labels need to be tested to ensure that they do not leach into the preparation through the plastic container. The way in which the drug is formulated can avoid some of the problems associated with oral administration. Drugs are usually taken orally as tablets or capsules. The drug (active substance) itself needs to be soluble in an aqueous solution at a controlled rate. Factors such as particle size and crystal form can significantly affect dissolution. Rapid dissolution is not always ideal. For example, a slow dissolution rate can result in a longer duration of action or can avoid an initial high plasma level.

[0269] In some embodiments, liposomes containing nanocarriers (e.g., liposomes containing an IDO prodrug) and / or an IDO prodrug, co-formulated with an immunomodulatory agent, are administered alone or in admixture with a physiologically acceptable carrier (such as saline or phosphate buffer) selected according to the route of administration and standard pharmaceutical practice. For example, when used as an injection, the nanocarrier can be formulated as a sterile suspension, dispersion or emulsion with a pharmaceutically acceptable carrier. In certain embodiments, normal saline can be used as the pharmaceutically acceptable carrier. Other suitable carriers include, for example, water, buffered water, 0.4% saline, 0.3% glycine, 5% glucose, etc., and glycoproteins for enhancing stability such as albumin, lipoproteins, globulins. In compositions containing saline or other salt-containing carriers, preferably the carrier is added after nanocarrier formation. Thus, after the nanocarrier is formed and loaded with the appropriate drug, the nanocarrier can be diluted in a pharmaceutically acceptable carrier such as normal saline. Similarly, IDO prodrug liposomes can be introduced into a carrier that facilitates suspension of the nanomaterial (e.g., emulsion, diluent, etc.).

[0270] The pharmaceutical composition can be sterilized by conventional well-known sterilization techniques. The resulting aqueous solution, suspension, dispersion, emulsion, etc. can be packaged for use or filtered under aseptic conditions. In certain embodiments, the drug delivery nanocarrier (e.g., nanoparticles coated with LB) is lyophilized and the lyophilized preparation is combined with a sterile aqueous solution prior to administration. The composition can also contain pharmaceutically acceptable auxiliary substances required to approximate physiological conditions, such as pH adjusters and buffers, isotonicity adjusters, etc., for example sodium acetate, sodium lactate, sodium chloride, potassium chloride, calcium chloride, etc.

[0271] Furthermore, in certain embodiments, the pharmaceutical formulation may include a lipid protecting agent that protects lipids from damage by free radicals and lipid peroxidation during storage. Lipophilic free radical scavengers such as α-tocopherol and water-soluble iron-specific chelating agents such as deferoxamine are suitable and contemplated herein. The concentration of the nanocarriers (e.g., liposomes containing an IDO prodrug) in the pharmaceutical formulation can vary widely, for example, less than about 0.05 wt%, usually at least about 2-5 wt% to 10-50 wt% or ~40 wt% or ~30 wt%, and is selected mainly according to the specific mode of administration, depending on factors such as fluid volume, viscosity, etc. For example, the concentration can be increased to reduce the fluid load associated with the treatment. This may be particularly desirable in patients with atherosclerotic congestive heart failure or severe hypertension. Alternatively, nanocarriers composed of irritating lipids can be diluted to a low concentration to reduce inflammation at the site of administration. The amount of nanocarriers administered depends on the specific drug used, the disease state being treated, and the judgment of the clinician, but generally ranges from about 0.01 to about 50 mg per kilogram of body weight, preferably from about 0.1 to about 5 mg per kilogram of body weight.

[0272] One of ordinary skill in the art will understand that the exact dosage will vary depending on factors such as the specific IDO prodrug and any co-formulated immunomodulatory agent and the desired medical effect, as well as patient factors such as age, gender, general condition, etc. One of ordinary skill in the art can use these factors to readily account for them and establish an effective therapeutic concentration without relying on undue experimentation.

[0273] For administration to a human (or non-human mammal) in a curative, palliative, retarding or prophylactic treatment of a disease described herein, the prescribing physician will ultimately determine the appropriate dosage of the drug for a given human (or non-human) subject, and the appropriate dosage can be expected to vary according to the age, weight and response of the individual, as well as the nature and severity of the patient's disease. In certain embodiments, the dosage of the drug provided by the nanocarrier can be approximately equal to the dosage used for the free drug. However, as noted above, the nanocarriers described herein can significantly reduce the toxicity of the drug administered by the nanocarrier and significantly increase the therapeutic window. Thus, in some cases, dosages in excess of those prescribed for the free drug are utilized.

[0274] One of ordinary skill in the art will appreciate and be enabled to make variations and modifications to the disclosed embodiments without departing from the functions and objectives of the invention disclosed herein. Such variations and modifications are intended to be within the scope of the present disclosure. IX.) Combination Therapy

[0275] As will be recognized and appreciated by one of ordinary skill in the art, the growth and survival of cancer cells can be affected by multiple signaling pathways. Thus, for treating such conditions, it is useful to combine different enzyme / protein / receptor inhibitors that exhibit different selectivities at the targets that regulate activity. Targeting more than one signaling pathway (or more than one biological molecule involved in a given signaling pathway) can reduce the likelihood of drug resistance occurring in a cell population and / or reduce the toxicity of the treatment.

[0276] Accordingly, liposomes containing the IDO prodrugs of the present disclosure can be used in combination with one or more other enzyme / protein / receptor inhibitors or one or more therapies for treating diseases such as cancer or infectious diseases. Examples of diseases and indications treatable by combination therapy include those described in the present disclosure. Examples of cancers include, but are not limited to, solid tumors and liquid tumors such as blood cancers. Examples of infectious diseases include viral infections, bacterial infections, fungal infections or parasitic infections.

[0277] For example, liposomes containing the IDO prodrugs of the present disclosure can be combined with one or more inhibitors of the following kinases for the treatment of cancer: Akt1, Akt2, Akt3, TGF-βR, PKA, PKG, PKC, CaM-kinase, phosphorylase kinase, MEKK, ERK, MAPK, mTOR, EGFR, HER2, HER3, HER4, INS-R, IGF-1R, IR-R, PDGFαR, PDGFβR, PI3K (α, β, γ, δ), CSFIR, KIT, FLK-II, KDR / FLK-1, FLK-4, flt-1, FGFR1, FGFR2, FGFR3, FGFR4, c-Met, Ron, Sea, TRKA, TRKB, TRKC, TAM kinases (Axl, Mer, Tyro3), FLT3, VEGFR / Flt2, Flt4, EphA1, EphA2, EphA3, EphB2, EphB4, Tie2, Src, Fyn, Lck, Fgr, Btk, Fak, SYK, FRK, JAK, ABL, ALK and B-Raf.

[0278] In a further embodiment, the liposomes containing the IDO prodrugs of the present disclosure can be combined with one or more of the following inhibitors for the treatment of cancer or infectious diseases. Non-limiting examples of inhibitors that can be combined with the compounds of the present disclosure for the treatment of cancer and infectious diseases include FGFR inhibitors (FGFR1, FGFR2, FGFR3 or FGFR4, such as INCB54828, INCB62079 and INCB63904), JAK inhibitors (JAK1 and / or JAK2, such as ruxolitinib, baricitinib or INCB39110), IDO inhibitors (such as epacadostat, NLG919 or BMS-986205), LSD1 inhibitors (such as INCB59872 and INCB60003), TDO inhibitors, PI3Kδ inhibitors (such as INCB50797 and INCB50465), PI3Kγ inhibitors, such as PI3Kγ-selective inhibitors, Pim inhibitors (such as INCB53914), CSF1R inhibitors, TAM receptor tyrosine kinases (Tyro-3, Axl and Mer), adenosine receptor antagonists (such as A2a / A2b receptor antagonists), HPK1 inhibitors, histone deacetylase inhibitors (HDAC), such as HDAC8 inhibitors, angiogenesis inhibitors, interleukin receptor inhibitors, bromo and extra-terminal family member inhibitors (such as bromodomain inhibitors or BET inhibitors, such as INCB54329 and INCB57643), poly ADP ribose polymerase (PARP) inhibitors, such as rucaparib, olaparib, niraparib, veliparib or talazoparib, arginase inhibitors (INCB01158), PD-1 inhibitors, PD-1 / L-1 inhibitors, PD-1 / L-2 inhibitors, and adenosine receptor antagonists, or combinations thereof.

[0279] In one embodiment, the A2a receptor inhibitor is the following prodrug compound (designated as "AR5"):

Chemical formula

[0280] In a further embodiment, the liposomes containing the IDO prodrug further comprise α-galactosylceramide (α-GalCer) (designated as "NK1").

[0281] Furthermore, the liposomes containing the IDO prodrug of the present disclosure can be further used in combination with other methods for treating cancer, such as chemotherapy, radiation therapy, tumor-targeted therapy, adjuvant therapy, immunotherapy or surgery.

[0282] Examples of immunotherapy include cytokine treatment (e.g., interferon, GM-CSF, G-CSF, IL-2), CRS-207 immunotherapy, cancer vaccines, monoclonal antibodies, adoptive T cell transfer, Toll receptor agonists, STING agonists, oncolytic virus therapy and immunomodulatory small molecules including thalidomide or JAK1 / 2 inhibitors.

[0283] Liposomes containing an IDO prodrug can be administered in combination with one or more anticancer agents such as chemotherapeutic agents. Examples of chemotherapeutic agents include abarelix, aldesleukin, alemtuzumab, alitretinoin, allopurinol, altretamine, anastrozole, arsenic trioxide, asparaginase, azacitidine, bevacizumab, bexarotene, baricitinib, bleomycin, bortezomib, bortezomib, busulfan intravenous, busulfan oral, calusterone, capecitabine, carboplatin, carmustine, cetuximab, chlorambucil, cisplatin, cladribine, clofarabine, cyclophosphamide, cytarabine, dacarbazine, dactinomycin, dalteparin sodium, dasatinib, daunorubicin, decitabine, denileukin, denileukin diftitox, dexrazoxane, docetaxel, doxorubicin, drostanolone propionate, eculizumab, epirubicin, erlotinib, estramustine, etoposide phosphate, etoposide, exemestane, fentanyl citrate, filgrastim, floxuridine, fludarabine, fluorouracil, fulvestrant, gefitinib, gemcitabine, gemtuzumab ozogamicin, goserelin acetate, histrelin acetate, ibritumomab tiuxetan, idarubicin, ifosfamide, imatinib mesylate, interferon α2a, irinotecan, lapatinib ditosylate, lenalidomide, letrozole, leucovorin, leuprolide acetate, levamisole, lomustine, mechlorethamine, megestrol acetate, melphalan, mercaptopurine, methotrexate, methoxsalen, mitomycin C, mitotane, mitoxantrone, nandrolone phenylpropionate, nelarabine, nolatrexed, olaparib, oxaliplatin, paclitaxel, pamidronate, panitumumab, pegaspargase, pegfilgrastim, pemetrexed disodium, pentostatin, pipobroman, plicamycin, procarbazine, quinacrine, rasburicase, rituximab, lurtotecan, lucaparib, sorafenib, streptozocin, sunitinib, sunitinib maleate, tamoxifen, temozolomide, teniposide, testolactone, thalidomide, thioguanine, thiotepa, topotecan,Examples include any of tamoxifen, tositumomab, trastuzumab, tretinoin, uracil mustard, barasertib, vinblastine, vincristine, vinorelbine, vorinostat, niraparib, veliparib, talazoparib, and zoledronic acid.

[0284] Other anti-cancer agents include antibody therapeutics such as trastuzumab (Herceptin), antibodies against co-stimulatory molecules such as CTLA-4 (e.g., ipilimumab), 4-1BB (e.g., urelumab, utomilumab), antibodies against PD-1 and PD-L1 / L2, or antibodies against cytokines (such as IL-10, TGF-β).

[0285] Examples of antibodies against PD-1 and / or PD-L1 / L2 that can be combined with the compounds of the present disclosure for the treatment of cancer or infectious diseases such as viral, bacterial, fungal, and parasitic infections include, but are not limited to, nivolumab, pembrolizumab, MPDL3280A, MEDI-4736, and SHR-1210.

[0286] Furthermore, liposomes containing the IDO prodrugs of the present disclosure can be used in combination with one or more immune checkpoint inhibitors for the treatment of diseases such as cancer or infectious diseases. Exemplary immune checkpoint inhibitors include inhibitors against immune checkpoint molecules such as CD27, CD28, CD40, CD122, CD96, CD73, CD47, OX40, GITR, CSF1R, JAK, PI3Kδ, PI3Kγ, TAM, arginase, CD137 (also known as 4-1BB), ICOS, A2AR, B7-H3, B7-H4, BTLA, CTLA-4, LAG3, TIM3, VISTA, PD-1, PD-L1, and PD-L2.

[0287] In some embodiments, the immune checkpoint molecule is a stimulatory checkpoint molecule selected from CD27, CD28, CD40, ICOS, OX40, GITR, and CD137. In further embodiments, the immune checkpoint molecule is an inhibitory checkpoint molecule selected from A2AR, B7-H3, B7-H4, BTLA, CTLA-4, IDO, KIR, LAG3, PD-1, TIM3, and VISTA. In further embodiments, the liposomes containing the IDO prodrugs provided herein can be used in combination with one or more agents selected from KIR inhibitors, TIGIT inhibitors, LAIR1 inhibitors, CD160 inhibitors, 2B4 inhibitors, and TGFRβ inhibitors. X.) Method of delivering liposomes containing an IDO prodrug to cells expressing IDO-1

[0288] As is known in the art, a variety of compositions and methods for using prodrugs and / or liposomes to kill tumor cells are known in the art. In the context of cancer, typical methods involve administering to a mammal having a tumor a biologically effective amount of an IDO prodrug of the present disclosure and / or a liposome of the present disclosure containing an IDO prodrug.

[0289] A typical embodiment is a method of delivering a therapeutic agent to cells expressing IDO-1, the method comprising forming an IDO prodrug by conjugating a drug moiety of the present disclosure to a lipid of the present disclosure via a linking unit, and exposing the cells to the IDO prodrug.

[0290] In one embodiment, the IDO prodrug comprises a drug moiety of Formula I conjugated via an LU comprising a hydroxymethylcarbamate linker and CHEMS.

[0291] In one embodiment, the IDO prodrug comprises a drug moiety of Formula I conjugated via an LU comprising a hydroxymethylcarbamate linker and stearic acid.

[0292] In one embodiment, the IDO prodrug comprises ID3 and CHEMS conjugated via a LU comprising a hydroxymethylcarbamate linker.

[0293] In one embodiment, the IDO prodrug comprises ID3 and stearic acid conjugated via a LU comprising a hydroxymethylcarbamate linker.

[0294] Another exemplary embodiment is a method of treating an individual suspected of having metastatic cancer, the method comprising parenterally administering to the individual a pharmaceutical composition comprising a therapeutically effective amount of an IDO prodrug produced by conjugating a drug moiety to a lipid of the present disclosure via a linking unit, and exposing cells to the IDO prodrug.

[0295] In one embodiment, the IDO prodrug comprises a drug moiety of Formula I and CHEMS conjugated via a LU comprising a hydroxymethylcarbamate linker.

[0296] In one embodiment, the IDO prodrug comprises a drug moiety of Formula I and stearic acid conjugated via a LU comprising a hydroxymethylcarbamate linker.

[0297] In one embodiment, the IDO prodrug comprises ID3 and CHEMS conjugated via a LU comprising a hydroxymethylcarbamate linker.

[0298] In one embodiment, the PD1 prodrug comprises ID3 and stearic acid conjugated via a LU comprising a hydroxymethylcarbamate linker.

[0299] The IDO prodrugs, liposomes, and co-formulated liposomes of the present disclosure inhibit the activity of IDO-1 protein / protein interactions and are thus useful in the treatment of diseases and disorders associated with the activity of IDO-1, as well as diseases and disorders associated with the kynurenine pathway, including interactions with other proteins such as IDO-2 and TDO. In further embodiments of the present disclosure, the IDO prodrugs, liposomes, or pharmaceutically acceptable salts or stereoisomers thereof are useful for therapeutic administration to enhance, stimulate, and / or increase immunity in cancer, chronic infection, or sepsis, including enhancing the response to vaccination.

[0300] In further embodiments, the present disclosure provides a method for inhibiting IDO-1 T cell function. The method includes administering to an individual or patient an IDO prodrug, liposome, and / or any of the formulas described herein (e.g., ID3), or as described in any of the claims, and the IDO prodrugs, liposomes, and nanoencapsulated IDO inhibitor prodrugs described herein, or pharmaceutically acceptable salts or stereoisomers thereof. The IDO prodrugs, liposomes, and nanoencapsulated IDO inhibitor prodrugs of the present disclosure can be used alone, in combination with other agents or therapies, or as immunostimulants or tumor immunostimulants for the treatment of diseases or disorders including cancer and other diseases. For the uses and methods described herein, any of the IDO prodrugs, liposomes, and nanoencapsulated IDO prodrugs of the present disclosure, including any of those embodiments, can be used.

[0301] Furthermore, the IDO prodrugs, liposomes, and nanoencapsulated IDO inhibitor prodrugs of the present disclosure inhibit the IDO-1 kynurenine pathway and / or T cell function, resulting in blockade of the IDO pathway. As is known in the art, IDO is activated during tumorigenesis and helps malignant cells escape eradication by the immune system. See MUNN et al., Trends in Immunology, 37(3)193-207(2016).

[0302] In a further embodiment, the present disclosure provides for the treatment of an individual or patient in vivo with an IDO prodrug, a liposome, and a nanoencapsulated IDO inhibitor prodrug, or salts or stereoisomers thereof, such that the growth of a cancerous tumor is inhibited.

[0303] The IDO prodrug, liposome, and nanoencapsulated IDO inhibitor prodrug, or any of the formulas described herein (e.g., ID3), or as described in any of the claims, and the IDO prodrug, liposome, and nanoencapsulated IDO inhibitor prodrug described herein, or salts or stereoisomers thereof, can be used to inhibit the growth of a cancerous tumor.

[0304] Alternatively, the IDO prodrug, liposome, and nanoencapsulated IDO prodrug of the present disclosure, or any of the formulas described herein, or as described in any of the claims, and the compounds described herein (e.g., ID3), or salts or stereoisomers thereof, can be used in combination with other agents or standard cancer treatments, as described in the present disclosure.

[0305] In a further embodiment, the present disclosure provides a method for inhibiting the growth of tumor cells in vitro. The method includes contacting the tumor cells in vitro with the IDO prodrug, liposome, and nanoencapsulated IDO inhibitor prodrug of the present disclosure, or any of the formulas described herein (e.g., ID3), or as described in any of the claims, and the IDO prodrug, liposome, and nanoencapsulated IDO inhibitor prodrug described herein, or salts or stereoisomers thereof.

[0306] In a further embodiment, the present disclosure provides a method for inhibiting the growth of tumor cells in a patient. The method comprises contacting the tumor cells with an IDO prodrug, a liposome and a nanoencapsulated IDO inhibitor prodrug of the present disclosure, or any of the formulas described herein (e.g., ID3), or any of the claims, and an IDO prodrug, a liposome and a nanoencapsulated IDO inhibitor prodrug described herein, or salts or stereoisomers thereof. XI.) Methods of treating cancer and other immune disorders

[0307] Another embodiment of the present disclosure is a method for treating cancer. The method comprises administering to a patient a therapeutically effective amount of a liposome comprising an IDO prodrug (i.e., ID3) herein, a compound described in any of the claims and described herein, or a salt thereof. Examples of cancers include cancers whose growth can be inhibited using the IDO inhibitors and IDO prodrugs of the present disclosure, as well as cancers that typically respond to immunotherapy.

[0308] In some embodiments, the present disclosure provides a method for enhancing, stimulating, and / or increasing an immune response in a patient. The method comprises administering to the patient a therapeutically effective amount of an IDO prodrug and / or a liposome comprising the same (i.e., ID3), a compound or composition described in any of the claims and described herein, or a salt thereof.

[0309] Non-limiting examples of cancers treatable using the liposomes comprising the IDO prodrugs of the present disclosure, the IDO prodrugs, and the co-formulated liposomes include bone cancer, pancreatic cancer, skin cancer, head and neck cancer, cutaneous or intraocular melanoma, uterine cancer, ovarian cancer, rectal cancer, cancer of the anal region, gastric cancer, testicular cancer, uterine cancer, fallopian tube carcinoma, endometrial carcinoma, endometrial cancer, cervical carcinoma, vaginal carcinoma, vulvar carcinoma, Hodgkin's disease, non-Hodgkin lymphoma, esophageal cancer, small intestine cancer, endocrine system cancer, thyroid cancer, parathyroid cancer, adrenal cancer, soft tissue sarcoma, urethral cancer, penile cancer, acute myeloid leukemia, chronic myeloid leukemia, acute lymphoblastic leukemia, chronic or acute leukemia including chronic lymphocytic leukemia, pediatric solid tumors, lymphocytic lymphoma, bladder cancer, kidney or urethral cancer, renal pelvis carcinoma, central nervous system (CNS) neoplasms, primary CNS lymphoma, tumor angiogenesis, spinal cord axis tumors, brainstem gliomas, pituitary adenomas, Kaposi's sarcoma, epidermoid cancer, squamous cell carcinoma, T-cell lymphoma, environmentally induced cancers including those induced by asbestos, and combinations of the foregoing cancers, but are not limited thereto. The compounds of the present disclosure are also useful for the treatment of metastatic cancers, particularly metastatic cancers that express IDO-1.

[0310] In some embodiments, cancers treatable with the liposomes or IDO prodrugs of the present disclosure include melanoma (e.g., metastatic malignant melanoma), renal cancer (e.g., clear cell carcinoma), prostate cancer (e.g., hormone-refractory adenocarcinoma of the prostate), breast cancer, colon cancer, lung cancer (e.g., non-small cell lung cancer and small cell lung cancer), squamous cell head and neck cancer, urothelial cancer (e.g., bladder), and cancers having high microsatellite instability (MSI high ). Further, the present disclosure includes refractory or recurrent malignancies whose growth can be inhibited using the liposomes or IDO prodrugs or co-formulated liposomes of the present disclosure.

[0311] In further embodiments, cancers treatable using the formulated and / or co-formulated liposomes or IDO prodrugs of the present disclosure include solid tumors (e.g., prostate cancer, colorectal cancer, esophageal cancer, endometrial cancer, ovarian cancer, uterine cancer, kidney cancer, liver cancer, pancreatic cancer, stomach cancer, breast cancer, lung cancer, head and neck cancer, thyroid cancer, glioblastoma, sarcoma, bladder cancer, etc.), blood cancers (e.g., lymphoma, acute lymphoblastic leukemia (ALL), acute myeloid leukemia (AML), chronic lymphocytic leukemia (CLL), chronic myeloid leukemia (CML), etc. leukemias, DLBCL, mantle cell lymphoma, non-Hodgkin lymphoma (including relapsed or refractory NHL and relapsed follicular), Hodgkin lymphoma or multiple myeloma) and combinations of said cancers, but are not limited thereto.

[0312] In further embodiments, cancers treatable using the formulated and / or co-formulated liposomes or IDO prodrugs of the present disclosure include cholangiocarcinoma, bile duct cancer, triple negative breast cancer, rhabdomyosarcoma, small cell lung cancer, leiomyosarcoma, hepatocellular carcinoma, Ewing sarcoma, brain cancer, brain tumor, astrocytoma, neuroblastoma, neurofibroma, basal cell carcinoma, chondrosarcoma, epitheloid sarcoma, eye cancer, fallopian tube cancer, gastrointestinal cancer, gastrointestinal stromal tumor, hairy cell leukemia, intestinal cancer, pancreatic islet cell cancer, oral cancer, mouth cancer, laryngeal cancer, larynx cancer, lip cancer, mesothelioma, cervical cancer, nasal cavity cancer, eye cancer, eye melanoma, pelvic cancer, rectal cancer, renal cell carcinoma, salivary gland cancer, paranasal sinus cancer, spinal cord cancer, tongue cancer, tubular carcinoma, urethral cancer, urethral cancer and ureteral cancer, but are not limited thereto.

[0313] Furthermore, in some embodiments, the formulated and / or co-formulated liposomes of the present disclosure, or IDO prodrugs can be used to treat sickle cell disease and sickle cell anemia.

[0314] Furthermore, in some embodiments, diseases and conditions treatable using the formulated and / or co-formulated liposomes, or IDO prodrugs, of the present disclosure include, but are not limited to, blood cancers, sarcomas, lung cancers, gastrointestinal cancers, genitourinary cancers, liver cancers, bone cancers, nervous system cancers, gynecological cancers, and skin cancers.

[0315] Exemplary blood cancers include lymphomas and acute lymphoblastic leukemia (ALL), acute myeloid leukemia (AML), acute promyelocytic leukemia (APL), chronic lymphocytic leukemia (CLL), chronic myeloid leukemia (CML) and other leukemias, diffuse large B-cell lymphoma (DLBCL), mantle cell lymphoma, non-Hodgkin lymphoma (including relapsed or refractory NHL and relapsed follicular), Hodgkin lymphoma, myeloproliferative disorders (e.g., primary myelofibrosis (PMF), polycythemia vera (PV), and essential thrombocythemia (ET)), myelodysplastic syndromes (MDS), T-cell acute lymphoblastic lymphoma (T-ALL), and multiple myeloma (MM).

[0316] Exemplary sarcomas include chondrosarcoma, Ewing sarcoma, osteosarcoma, rhabdomyosarcoma, angiosarcoma, fibrosarcoma, liposarcoma, myxosarcoma, rhabdomyxosarcoma, rhabdomyoma, fibroma, lipoma, hamartoma, and teratoma.

[0317] Exemplary lung cancers include non-small cell lung cancer (NSCLC), small cell lung cancer, bronchogenic carcinoma (squamous, undifferentiated small cell, undifferentiated large cell, adenocarcinoma), alveolar (bronchiolar) carcinoma, bronchial adenoma, chondroid hamartoma, and mesothelioma.

[0318] Exemplary gastrointestinal cancers include cancers of the esophagus (squamous cell carcinoma, adenocarcinoma, leiomyosarcoma, lymphoma), stomach (carcinoma, lymphoma, leiomyosarcoma), pancreas (ductal adenocarcinoma, insulinoma, glucagonoma, gastrinoma, carcinoid tumor, VIP-producing tumor), small intestine (adenocarcinoma, lymphoma, carcinoid tumor, Kaposi sarcoma, leiomyosarcoma, hemangioma, lipoma, neurofibroma, fibroma), large intestine (adenocarcinoma, tubular adenoma, villous adenoma, hamartoma, leiomyosarcoma), and colorectal cancer.

[0319] Exemplary genitourinary cancers include cancers of the kidney (adenocarcinoma, Wilms' tumor [nephroblastoma]), bladder and urethra (squamous cell carcinoma, transitional cell carcinoma, adenocarcinoma), prostate (adenocarcinoma, sarcoma) and testis (seminoma, teratoma, embryonal carcinoma, teratocarcinoma, choriocarcinoma, sarcoma, interstitial cell carcinoma, fibroma, fibroadenoma, adenomatoid tumor, lipoma).

[0320] Exemplary liver cancers include hepatoma (hepatocellular carcinoma), cholangiocarcinoma, hepatoblastoma, angiosarcoma, hepatocellular adenoma and hemangioma.

[0321] Exemplary bone cancers include, for example, osteogenic sarcoma (osteosarcoma), fibrosarcoma, malignant fibrous histiocytoma, chondrosarcoma, Ewing's sarcoma, malignant lymphoma (reticulosarcoma), multiple myeloma, malignant giant cell tumor, chordoma, osteochondroma (osteochondral exostosis), benign chondroma, chondroblastoma, chondromyxofibroma, osteoid osteoma and giant cell tumor.

[0322] Exemplary cancers of the nervous system include cancers of the skull (osteoma, hemangioma, granuloma, xanthoma, Paget's disease), meninges (meningioma, meningiosarcoma, gliomatosis), brain (astrocytoma, medulloblastoma, glioma, ependymoma, germ cell tumor (pinealoma), glioblastoma, glioblastoma multiforme, oligodendroglioma, schwannoma, retinoblastoma, congenital tumor), and spinal cord (neurofibroma, meningioma, glioma, sarcoma), as well as neuroblastoma and Lhermitte - Duclos disease.

[0323] Exemplary gynecologic cancers include cancers of the uterus (endometrial carcinoma), cervix (cervical carcinoma, pre - tumor cervical dysplasia), ovary (ovarian cancer (serous cystadenocarcinoma, mucinous cystadenocarcinoma, unclassified carcinoma), granulosa - theca cell tumor, Sertoli - Leydig cell tumor, undifferentiated germ cell tumor, malignant teratoma), vulva (squamous cell carcinoma, intraepithelial carcinoma, adenocarcinoma, fibrosarcoma, melanoma), vagina (clear cell carcinoma, squamous cell carcinoma, sarcoma botryoides (fetal rhabdomyosarcoma)) and fallopian tube (carcinoma).

[0324] Exemplary skin cancers include melanoma, basal cell carcinoma, squamous cell carcinoma, Kaposi's sarcoma, dysplastic nevus, lipoma, hemangioma, dermatofibroma, and keloid. In some embodiments, diseases and conditions treatable using the compounds of the present disclosure include, but are not limited to, sickle cell disease (e.g., sickle cell anemia), triple negative breast cancer (TNBC), myelodysplastic syndrome, testicular cancer, cholangiocarcinoma, esophageal cancer, and urothelial cancer.

[0325] Furthermore, the formulations and / or co-formulations of liposomes of the present disclosure, or the blockade of the IDO-1 and / or kynurenine pathway by IDO prodrugs, can also be used to treat infectious diseases such as viral, bacterial, fungal, and parasitic infections.

[0326] The present disclosure provides a method for treating infectious diseases such as viral infections. This method includes administering to a patient a therapeutically effective amount of a liposome or IDO prodrug formulated and / or co-formulated as described in any of the claims and described herein, or any of the formulas described herein (i.e., ID3), and salts thereof.

[0327] Examples of viruses that cause infectious diseases treatable by the methods of the present disclosure include, but are not limited to, human immunodeficiency virus, human papillomavirus, influenza, hepatitis A, B, C or D virus, adenovirus, poxvirus, herpes simplex virus, human cytomegalovirus, severe acute respiratory syndrome virus, Ebola virus and measles virus. In some embodiments, viruses that cause infectious diseases treatable by the methods of the present disclosure include hepatitis (type A, B or C), herpes virus (e.g., VZV, HSV-1, HAV-6, HSV-II and CMV, Epstein-Barr virus), adenovirus, influenza virus, flavivirus, echovirus, rhinovirus, coxsackievirus, coronavirus, respiratory syncytial virus, mumps virus, rotavirus, measles virus, rubella virus, parvovirus, vaccinia virus, HTLV virus, dengue virus, papillomavirus, molluscum contagiosum virus, poliovirus, rabies virus, JC virus and arbovirus encephalitis virus, but are not limited thereto.

[0328] Furthermore, the present disclosure provides a method for treating a bacterial infection. The method includes administering to a patient a therapeutically effective amount of a liposome or IDO prodrug formulated and / or co-formulated as described in any of the claims and as described herein, or any of the formulas described herein (i.e., ID3), or salts thereof.

[0329] Examples of pathogenic bacteria that cause infectious diseases treatable by the methods of the present disclosure include, but are not limited to, Chlamydia, Rickettsia bacteria, Mycobacteria, Staphylococci, Streptococci, Pneumococci, Meningococci and Gonococci, Klebsiella, Proteus, Serratia, Pseudomonas, Legionella, Diphtheria, Salmonella, Bacilli, Cholera, Tetanus, Botulism, Anthrax, Plague, Leptospirosis, and Lyme disease bacteria.

[0330] Furthermore, the present disclosure provides a method for treating a fungal infection. The method comprises administering to a patient a therapeutically effective amount of a liposome or IDO prodrug formulated and / or co-formulated as described in any of the claims and as described herein, or any of the formulas described herein (i.e., ID3), or salts thereof.

[0331] Examples of pathogenic fungi that cause infectious diseases treatable by the methods of the present disclosure include, but are not limited to, Candida (albicans, krusei, glabrata, tropicalis, etc.), Cryptococcus neoformans, Aspergillus (fumigatus, Niger, etc.), Mucorales (Mucor, Absidia, Rhizopus), Sporothrix schenkii, Blastomyces dermatitidis, Paracoccidioides brasiliensis, Coccidioides immitis, and Histoplasma capsulatum.

[0332] Furthermore, the present disclosure provides a method for treating a parasitic infection. The method comprises administering to a patient a therapeutically effective amount of a liposome or IDO prodrug formulated and / or co-formulated as described in any of the claims and as described herein, or any of the formulas described herein (i.e., ID3), or salts thereof.

[0333] Examples of pathogenic parasites that cause infectious diseases treatable by the methods of the present disclosure include, but are not limited to, Entamoeba histolytica, Balantidium coli, Naegleriafowleri, Acanthamoeba sp., Giardia lambia, Cryptosporidium sp., Pneumocystis carinii, Plasmodium vivax, Babesia microti, Trypanosoma brucei, Trypanosoma cruzi, Leishmania donovani, Toxoplasma gondi, and Nippostrongylus brasiliensis.

[0334] In a further set of embodiments falling within the scope of the present disclosure, formulated and / or co-formulated liposomes, or IDO prodrugs, or any of the formulas described herein (i.e., ID3) are useful in preventing or reducing the risk of developing any of the diseases mentioned in the present disclosure, for example, in individuals who may have a predisposition to a disease, condition, or disorder but have not yet experienced or manifested the disease state or overall symptoms of the disease, in preventing or reducing the risk of developing the disease, condition, or disorder. XII.) Kit / Manufactured Product

[0335] For use in the laboratories, prognoses, prophylaxis, diagnosis and therapeutic applications described herein, the kits are within the scope of the present invention. Such kits can include a carrier, package or container compartmentalized to receive one or more containers such as vials, tubes, etc., each of the containers including a label or insert containing instructions for use, such as the uses described herein, together with one of the separate elements to be used in the method. For example, the container can include formulated and / or co-formulated liposomes that are detectably labeled or can be detectably labeled and / or loaded with an IDO prodrug of the present disclosure. The kit can include a container containing a drug unit. The kit can include all or part of the formulated and / or co-formulated liposomes and / or IDO prodrug.

[0336] The kits of the present invention typically include the above containers and one or more other containers associated with the above containers equipped with utensils desirable from a commercial and user perspective, such as buffers, diluents, filters, needles, syringes, etc., a label on a carrier, package, container, vial and / or tube listing the contents and / or instructions for use, and an accompanying document having instructions for use.

[0337] Labels can be present on or with a container to indicate that the composition is used for a particular therapeutic or non-therapeutic use, such as prognostic, prophylactic, diagnostic or laboratory use, and can also indicate instructions for in vivo or in vitro use, such as those described herein. Instructions and / or other information can also be included on an insert or label included with or on the kit. The label can be present on the container or can be associated with the container. When the letters, numbers or other characters forming the label are molded or etched into the container itself, the label can be present on the container, and when the label is present in a receptacle or carrier that also holds the container, such as an accompanying document, the label can be associated with the container. The label can indicate that the composition is used for diagnosing, treating, preventing or prognosticating a condition such as cancer or other immune disorders.

[0338] The terms "kit" and "manufactured article" can be used synonymously.

[0339] In another embodiment of the invention, manufactured articles containing formulations and / or co-formulations of liposomes and / or IDO prodrugs and the like are within the scope of this disclosure. Manufactured articles typically include at least one container and at least one label. Suitable containers include, for example, bottles, vials, syringes and test tubes. The container can be formed from a variety of materials such as glass, metal or plastic. The container can hold a formulation and / or co-formulation of liposomes loaded with an IDO prodrug.

[0340] Alternatively, the container can hold a composition effective to treat, diagnose, prognose or prevent a condition and can have a sterile access port (e.g., the container can be a vial having a stopper pierceable by an intravenous solution bag or a hypodermic needle). The active agent in the composition can be a liposome formulated and / or co-formulated with an IDO prodrug disclosed herein and / or an IDO prodrug.

[0341] The product can further include a second container containing a pharmaceutically acceptable buffer such as phosphate buffered saline, Ringer's solution and / or dextrose solution. The product can further include other articles desirable from a commercial and user standpoint, including other buffers, diluents, filters, stirrers, needles, syringes and / or package inserts with instructions and / or directions for use.

[0342] Exemplary Embodiments Among the embodiments provided are the following. 1) An IDO prodrug composition, wherein the IDO prodrug composition comprises (i) a drug moiety, (ii) a lipid moiety, (iii) a linking unit ("LU") and wherein the drug moiety comprises an IDO antagonist and the LU conjugates the drug moiety to the lipid moiety, an IDO prodrug composition.

[0343] 2) The IDO prodrug according to claim 1, further comprising the chemical structure set forth in Formula I.

[0344] 3) The IDO prodrug according to claim 1, wherein the drug moiety comprises the chemical structure shown as ID3.

[0345] 4) The IDO prodrug according to claim 1, wherein the LU is a hydroxymethylcarbamate linker.

[0346] 5) The IDO prodrug according to claim 1, wherein the lipid moiety comprises the lipid described in Table I.

[0347] 6) The IDO prodrug according to claim 1, wherein the lipid moiety comprises the lipid described in Table III.

[0348] 7) The IDO prodrug according to claim 1, wherein the lipid moiety comprises CHEMS.

[0349] 8) The IDO prodrug according to claim 1, wherein the lipid moiety comprises stearic acid.

[0350] 9) An IDO prodrug composition, wherein the IDO prodrug composition comprises (i) a drug moiety comprising ID3, (ii) a lipid moiety comprising CHEMS, (iii) a linker (LU) comprising a hydroxymethylcarbamate linker and is an IDO prodrug composition.

[0351] 10) An IDO prodrug composition, wherein the IDO prodrug composition comprises (i) a drug moiety comprising ID3, (ii) a lipid moiety comprising stearic acid, (iii) a linker (LU) comprising a hydroxymethylcarbamate linker and is an IDO prodrug composition.

[0352] 11) A liposome comprising an IDO prodrug, wherein the liposome releases an active IDO inhibitor after cleavage of the linker (LU).

[0353] 12) The liposome according to claim 11, wherein the linker (LU) is a hydroxymethylcarbamate linker.

[0354] 13) The liposome according to claim 11, further comprising a helper lipid, wherein the helper lipid is shown in Table II.

[0355] 14) The liposome according to claim 11, wherein the IDO prodrug comprises ID3.

[0356] 15) The liposome according to claim 11, further co-formulated with an immunomodulatory agent, wherein the immunomodulatory agent is selected from the group consisting of immunogenic cell death-inducing chemotherapeutic agents, toll-like receptor agonists, STING agonists, CTLA-4 inhibitors, PD-1 / PD-L1 inhibitors and / or prodrugs thereof.

[0357] 16) The liposome according to claim 11, further co-formulated with an ICD-inducing chemotherapeutic agent, wherein the ICD-inducing chemotherapeutic agent is selected from the group consisting of DOX, MTO, OXA, CP, bortezomib, carfilzomib or paclitaxel.

[0358] 17) The liposome according to claim 11, further comprising DOX.

[0359] 18) The liposome according to claim 11, further comprising MTO.

[0360] 19) The liposome according to claim 11, further comprising DOX.

[0361] 20) The liposome according to claim 11, further comprising MTO.

[0362] 21) A kit comprising the liposome according to claim 11.

[0363] 22) A kit comprising the liposome according to claim 15.

[0364] 23) A kit comprising the liposome according to claim 16.

[0365] 24) A method of treating a subject suffering from or diagnosed with cancer, comprising: (i) administering to a subject in need thereof an effective amount of liposomes, said liposomes comprising an IDO prodrug; (ii) its pharmaceutically acceptable salts; A method comprising.

[0366] 25) The method according to claim 24, wherein the IDO prodrug comprises ID3.

[0367] 26) The method according to claim 24, wherein the liposomes further comprise ID3 co-formulated with an ICD-inducing chemotherapeutic agent.

[0368] 27) The method according to claim 24, wherein the liposomes further comprise ID3 co-formulated with an immunomodulatory agent.

[0369] 28) The liposome according to claim 11, wherein the IDO prodrug comprises ID3 co-formulated with AR5.

[0370] 29) The liposome according to claim 11, wherein the IDO prodrug comprises ID3 co-formulated with AR5 and TR5.

[0371] 30) The liposome according to claim 11, wherein the IDO prodrug comprises ID3 co-formulated with NK1.

[0372] 31) The liposome according to claim 11, wherein the IDO prodrug comprises ID3 co-formulated with NK1 and MTO.

[0373] 32) The liposome according to claim 11, wherein the IDO prodrug comprises ID3 co-formulated with TR3.

[0374] 33) The liposome according to claim 11, wherein the IDO prodrug comprises ID3 co-formulated with TR5.

[0375] 34) The liposome according to claim 11, wherein the IDO prodrug comprises ID3 co-formulated with TB4.

[0376] 35) The liposome according to claim 11, wherein the IDO prodrug comprises ID3 co-formulated with PD3.

[0377] 36) The liposome according to claim 11, wherein the IDO prodrug comprises ID3 co-formulated with AR5 and TR3.

[0378] 37) The liposome according to claim 11, wherein the IDO prodrug comprises ID3 co-formulated with AR5 and TB4.

[0379] 38) The liposome according to claim 11, wherein the IDO prodrug comprises ID3 co-formulated with AR5 and PD3.

Examples

[0380] Various aspects of the present invention are further illustrated and exemplified by several of the following examples, none of which are intended to limit the scope of the present invention.

[0381] Example 1: General Chemical Synthesis of a Protected ID3 Prodrug Containing Cholesterol Hemisuccinate (「CHEMS」) The chemical synthesis of the protected ID3 prodrug containing CHEMS is synthesized using the following protocol. First, quinoline (1) and boronate (2) (BARLIND, J.G. et al., J. Med. Chem., 55:10610 - 10629 (2012)) were coupled with cesium carbonate and PEPPSI-IPr catalyst in dioxane / water at 100 °C to obtain intermediate 3. Then, intermediate 3 was hydrogenated over palladium carbon to obtain intermediate 4. Next, intermediate 4 was hydrolyzed with lithium hydroxide in aqueous ethanol solution at 50 °C to obtain intermediate 5. Next, in THF, intermediate 5 was coupled with chiral auxiliary reagent (6) using triethylamine and pivaloyl chloride, and the diastereomers were separated to obtain intermediate 7. Then, intermediate 7 was alkylated with methyl iodide and NaHMDS in THF at -50 °C to obtain intermediate 8. Next, the chiral auxiliary was cleaved from intermediate 8 using LiOH and hydrogen peroxide in aqueous THF solution at 0 °C to obtain intermediate 9. Then, in ethyl acetate, intermediate 9 was coupled with aniline (10) using polyphosphonic anhydride and pyridine to obtain intermediate 11. Next, intermediate 11 was treated with reagent (12) in THF and then with sodium iodide to obtain intermediate 13. Finally, it was refluxed in THF and treated with silver salt of CHEMS (14) to obtain the final ID3 prodrug. (Figure 1).

[0382] Example 2: Chemical synthesis of ID3 prodrug intermediate. The (if) chemical synthesis of the ID3 prodrug was carried out in the following manner. To a solution of the ID3 drug moiety (6.00 g, 14.6 mmol, 1.00 equiv) in THF (60 mL) was added LiHMDS (1 M, 29.2 mL, 2.00 equiv) at -70 °C, and the reaction mixture was stirred at -70 °C for 0.5 h. Then, a solution of compound 1 (3.24 g, 25.1 mmol, 2.23 mL, 1.72 equiv) in THF (10 mL) was added to the mixture at -78 °C, and the mixture was stirred at -78 °C for an additional 1 h. TLC (petroleum ether:ethyl acetate = 2:1) showed that the ID3 drug moiety (R f = 0.2) was consumed and one major new spot (R fIt was shown that (=0.6) was formed. The reaction mixture was poured into a saturated aqueous NH4Cl solution (150 mL) and extracted with ethyl acetate (100 mL × 2). The combined organic layers were washed with brine (100 mL), dried over Na2SO4, filtered, and concentrated to obtain crude compound 2 (7.60 g, unpurified) as a yellow oil. The obtained compound is shown in Figure 2.

[0383] Example 3: Alternative chemical synthesis of ID3 prodrug intermediate. In another experiment, an alternative chemical synthesis of the ID3 prodrug (if) was carried out in the following manner. To a solution of the ID3 drug moiety (3.00 g, 7.30 mmol, 1.00 equiv) in THF (30.0 mL) was added LiHMDS (1 M, 14.6 mL, 2.00 equiv) at -70 °C. The reaction mixture was then stirred at -70 °C for 0.5 h. Then, a solution of compound 3a (6.57 g, 51.0 mmol, 4.53 mL, 6.98 equiv) in THF (15.0 mL) was added to the mixture at -70 °C and stirred for an additional 1 h. 1 1H NMR showed that the ID3 drug moiety was correct. LCMS showed that the drug moiety was completely consumed and the desired MS (RT = 0.877 min) was detected. The mixture was poured into a saturated solution of NH4Cl (200 mL) and extracted with ethyl acetate (200 mL × 3). The combined organics were washed with brine (200 mL), dried over Na2SO4, filtered, and concentrated to obtain a crude product. Compound 3 (4.00 g, unpurified) was obtained as a yellow oil. The obtained compound is shown in Figure 3.

[0384] Example 4: Chemical synthesis of ID3 prodrug containing stearic acid In another experiment, an ID3 prodrug containing stearic acid was synthesized in the following manner. To a solution of stearic acid (3.39 g, 11.9 mmol, 4.01 mL, 0.80 equivalent) in DMF (400 mL) was added Ag2CO3 (6.16 g, 22.3 mmol, 1.01 mL, 1.50 equivalents) at 25 °C. The reaction mixture was stirred at 25 °C for 0.5 h. Subsequently, compound 2 (Figure 2) (7.50 g, 14.9 mmol, 1.00 equivalent) and NaI (3.35 g, 22.3 mmol, 1.50 equivalents) were added to the mixture. After the addition, the reaction mixture was stirred at 80 °C for an additional 12 h. LCMS indicated that the reaction was complete and the desired mass (RT = 1.309 min) was detected. The reaction mixture was cooled to 25 °C, filtered through a pad of celite, and washed with DCM (300 mL). The filtrate was concentrated at 50 °C to obtain the crude product. The crude product was purified by reverse-MPLC (TFA conditions) and then concentrated under reduced pressure to obtain the crude product. The crude product was purified by column chromatography (SiO2, petroleum ether:ethyl acetate = 5:1 to 2:1, R f = 0.5), and when detected by TLC (petroleum ether:ethyl acetate = 2:1, R f = 0.5, PMA), (2.36 g, 3.09 mmol, yield 20.7%, purity 98.3%) was obtained as a yellow solid, which was confirmed by 1 1H NMR, 19 19F NMR, LCMS, HPLC, and SFC. The resulting compound is shown in Figure 7.

[0385] Example 5: Chemical synthesis of an ID3 prodrug containing cholesterol hemisuccinate ("CHEMS"). In another experiment, an ID3 prodrug containing cholesterol hemisuccinate ("CHEMS") was synthesized in the following manner. Briefly, a mixture of cholesterol hemisuccinate (5.80 g, 11.9 mmol, 1.50 equiv) and Ag2CO3 (3.29 g, 11.9 mmol, 540 μL, 1.50 equiv) in DMF (15.0 mL) was stirred at 25 °C for 0.5 h. To the mixture was added compound 3 (Figure 3) (4.00 g, 7.95 mmol, 1.00 equiv) and NaI (1.43 g, 9.54 mmol, 1.20 equiv) in DMF (15.0 mL) at 25 °C. The mixture was stirred at 80 °C for 16 h. LCMS indicated that compound 3 was completely consumed and the desired MS (RT = 1.286 min) was detected. The mixture was filtered and the filtrate was collected. The filtrate was concentrated to obtain the crude product. Purification of the crude product by column chromatography (silica gel, ethyl acetate / petroleum ether = 1 / 10 - 1 / 4) gave the major spot (petroleum ether / ethyl acetate = 2 / 1, R f = 0.4). The unpurified compound (target 2) (3.00 g, 3.15 mmol, 39.6% yield) was obtained as a pale yellow solid. The obtained compound is shown in Figure 8.

[0386] Example 6: Synthesis and Characterization of LNP-ID3 Liposomes In another experiment, liposomes containing the ID3 prodrug (denoted as LNP-ID3) were synthesized as follows. Briefly, the stock solutions of each lipid component of LNP-ID3 were prepared in ethanol at a concentration of 20 mg / ml. Thus, the ethanolic stock solutions of Hydrogenated Soy PC [(HSPC: L-α-phosphatidylcholine, hydrogenated (soybean))], cholesterol, and DSPE-PEG (1,2-distearoyl-sn-glycero-3-phosphoethanolamine-N-[amino(polyethylene glycol)-2000]) were prepared at the above concentrations. The stock solution of the ID3 prodrug (20 mg / ml) was prepared in acetonitrile. Each lipid component (HSPC, CHOL, DSPE-PEG, and ID3 prodrug) was mixed at a molar ratio of 51:27:17:5 to synthesize LNP-ID3. The size of the liposome LNP-ID3 depends on various parameters such as (i) flow rate, (ii) temperature, and (iii) concentration of the lipid mixture. The optimized ratio of the lipid mixture at a molar ratio of 51:27:17:5 was preheated at 50 °C. The aqueous phase containing 1 mM PBS buffer was also preheated at 50 °C before passing through a microfluidics cartridge (Precision NanoSystems, Inc.) at a flow rate of 3:1 (aqueous phase: organic phase, lipid mixture). The solvent was removed using a dialysis membrane (Sigma Aldrich) with a 12KDa size cutoff against DI water for at least 24 hours. To optimize the removal of the solvent, the dialysis water was changed at least 5 times during the 24-hour period. After removing the solvent, LNP-ID3 was concentrated using an Amicon centrifugal filtration device (size 10 kDa, 3000g).

[0387] The properties of the LNP-ID3 liposomes were determined using a Malvern Zetasizer (Malvern Instrumentation Co., Westborough, MA, USA). Briefly, 2 ml of LNP-ID3 liposomes (the concentration of the liposomes was 0.5 - 1 mg / ml) was placed in a four-sided transparent plastic cuvette and analyzed directly at 25 °C. The results shown in Figure 9 indicate that the Zav size of the nanoparticles was approximately 80 nm and the PDI was approximately 0.203.

[0388] Furthermore, the zeta potential of LNP-ID3 liposomes in the aqueous dispersion was determined using a Malvern zeta seizer Instrument (Malvern Instrumentation Co, Westborough, MA, USA). Briefly, approximately 1 ml of liposomes (concentration approximately 2 mg / ml in 20 mM NaCl) was placed into a disposable capillary zeta potential cell available for the Zetasizer. The measurement was carried out at 25 °C. The results indicate that the zeta potential determination of LNP-ID3 was approximately -11.6 mV (Figure 10).

[0389] Example 7: Synthesis and Characterization of LNP-AR5-ID3 Liposomes In another experiment, liposomes containing the ID3 prodrug and the A2a receptor inhibitor prodrug (AR5) (designated as LNP-AR5-ID3) were synthesized and characterized as follows. Briefly, they were synthesized with a molar ratio of HSPC:CHOL:AR5:ID3:DSPE-PEG of 52:27:8:8:5, fixing the molar ratio of AR5:ID3 at 1:1. The optimized ratio of the lipid mixture at a molar ratio of 52:27:8:8:5 was preheated at 55 - 60 °C. The final concentration of the lipid mixture was 2.5 mg / ml. Before passing through a microfluidics cartridge (Precision NanoSystems, Inc.) at a flow rate of 3:1 (aqueous phase: organic phase, lipid mixture), the aqueous phase containing 1 mM PBS buffer was also preheated at 55 - 60 °C. The solvent was removed for at least 24 hours using a dialysis membrane (Sigma Aldrich) that removes 12KDa size against DI water. To optimize the removal of the solvent, the dialysis water was exchanged at least 5 times during the 24-hour period. After removing the solvent, LNP-AR5-ID3 was concentrated using an Amicon centrifugal filtration device (cut-off size 10 kDa, 3000 g).

[0390] The properties of the LNP-AR5-ID3 liposomes were determined using a Malvern Zetasizer (Malvern Instrumentation Co., Westborough, MA, USA). Briefly, 2 ml of LNP-AR5-ID3 liposomes (liposome concentration was 0.5 - 1 mg / ml) were placed in a four-sided transparent plastic cuvette and analyzed directly at 25°C. The results shown in Figure 11 indicate that the Zav size of the nanoparticles was approximately 91 nm and the PDI was approximately 0.212.

[0391] Furthermore, the zeta potential of the LNP-AR5-ID3 liposomes in aqueous dispersion was determined using a Malvern zeta seizer Instrument (Malvern Instrumentation Co, Westborough, MA, USA). Briefly, approximately 1 ml of liposomes (concentration approximately 2 mg / ml in 20 mM NaCl) was placed in a disposable capillary zeta potential cell available for the Zetasizer. The measurement was carried out at 25°C. The results indicate that the zeta potential determination of LNP-AR5-ID3 was approximately -14.1 mV (Figure 12).

[0392] Example 8: Synthesis and property determination of LNP-AR5-TR5-ID3 liposomes In another experiment, liposomes containing an ID3 prodrug, an A2a receptor inhibitor prodrug (AR5), and telratrimod (TR5) (designated as LNP-AR5-TR5-ID3) were synthesized and characterized as follows. Briefly, a lipid stock solution of HSPC, CHOL, DSPE-PEG, and the AR5 prodrug. TR5 was prepared separately in ethanol (20 mg / ml). Also, a stock solution of the ID3 prodrug was prepared in acetonitrile (20 mg / ml). HSPC:CHOL:AR5:TR5:ID3:DSPE-PEG was synthesized using a molar ratio of 51.6:27:7:2.4:7:5. The optimized ratio of the lipid mixture at a molar ratio of 51.6:27:7:2.4:7:5 was preheated at 55 - 60 °C. The final concentration of the prodrug AR5 in the lipid mixture was 0.625 mg / ml. Before passing through a microfluidics cartridge (Precision NanoSystems, Inc.) at a flow rate of 4.5:1 (aqueous phase: organic phase, lipid mixture), the aqueous phase containing 1 mM PBS buffer was also preheated at 55 - 60 °C. The solvent was removed using a dialysis membrane (Sigma Aldrich) with a 12KDa size cut-off against DI water for at least 24 hours. To optimize the removal of the solvent, the dialysis water was changed at least 5 times during the 24-hour period. After removing the solvent, LNP-AR5-TR5-ID3 was concentrated using an Amicon centrifugal filtration device (cut-off size 10 kDa, 3000 g).

[0393] The properties of the LNP-AR5-TR5-ID3 liposomes were determined using a Malvern Zetasizer (Malvern Instrumentation Co., Westborough, MA, USA). Briefly, 2 ml of LNP-AR5-TR5-ID3 liposomes (the concentration of the liposomes was 0.5 - 1 mg / ml) were placed in a four-sided transparent plastic cuvette and analyzed directly at 25 °C. The results shown in Figure 13 indicate that the Zav size of the nanoparticles was approximately 96 nm and the PDI was approximately 0.117.

[0394] Furthermore, the zeta potential of LNP-AR5-TR5-ID3 liposomes in the aqueous dispersion was determined using a Malvern zeta seizer Instrument (Malvern Instrumentation Co, Westborough, MA, USA). Briefly, approximately 1 ml of liposomes (concentration approximately 2 mg / ml in 20 mM NaCl) was placed into a disposable capillary zeta potential cell available for the Zetasizer. The measurement was performed at 25 °C. The results indicate that the zeta potential determination of LNP-AR5-TR5-ID3 was approximately -20.5 mV (Figure 14).

[0395] Example 9: Synthesis and Characterization of LNP-ID3-NK1 Liposomes In another experiment, liposomes containing the ID3 prodrug and α-galactosylceramide (α-GalCer) (NK1) (designated as LNP-ID3-NK1) were synthesized and characterized as follows. Briefly, a stock solution of the ID3 prodrug (20 mg / ml) was prepared in acetonitrile, and a stock solution of NK1 (10 mg / ml) was prepared in DMSO. A lipid mixture of HSPC, CHOL, ID3, NK1, and DSPE-PEG in a molar ratio of 51:29:16:0.085:4 was prepared by mixing appropriate amounts of all the lipid stock solutions and then further diluting with ethanol to obtain a lipid concentration of 10 mg / ml. This lipid mixture was preheated at 50 °C using a microfluidizer. Similarly, the aqueous phase containing 1 mM PBS buffer was also preheated at 50 °C before passing through a microfluidic cartridge (Precision NanoSystems, Inc.) at a flow rate of 3:1 (aqueous phase: organic phase, lipid mixture). Thus, the molar ratio of ID3:NK1 remained 12:1. The solvent was removed using a dialysis membrane (Sigma Aldrich) with a 12 kDa size exclusion for DI water for at least 24 hours. To optimize the removal of the solvent, the dialysis water was changed at least 5 times during the 24-hour period. After removing the solvent, LNP-ID3-NK1 was concentrated using an Amicon centrifugal filtration device (cut-off size 10 kDa, 3000 g).

[0396] The properties of LNP-ID3-NK1 liposomes were determined using a Malvern Zetasizer (Malvern Instrumentation Co., Westborough, MA, USA). Briefly, 2 ml of LNP-ID3-NK1 liposomes (liposome concentration was 0.5 - 1 mg / ml) were placed in a four-sided transparent plastic cuvette and analyzed directly at 25°C. The results shown in Figure 15 indicate that the Zav size of the nanoparticles was approximately 80 nm and the PDI was approximately 0.189.

[0397] Furthermore, the zeta potential of LNP-ID3-NK1 liposomes in an aqueous dispersion was determined using a Malvern zeta seizer Instrument (Malvern Instrumentation Co, Westborough, MA, USA). Briefly, approximately 1 ml of liposomes (concentration approximately 2 mg / ml in 20 mM NaCl) was placed in a disposable capillary zeta potential cell available for the Zetasizer. The measurement was carried out at 25°C. The results indicate that the zeta potential determination of LNP-ID3-NK1 was approximately -13.1 mV (Figure 16).

[0398] Example 10: Synthesis and Characterization of Mitochondrial-Loaded LNP-ID3-NK1 Liposomes In another experiment, LNP-ID3-NK1 was loaded into liposomes containing mitoxantrone (MTO) (designated as LNP-ID3-NK1-MTO), synthesized and characterized as follows. Briefly, a stock solution of MTO (0.6 mg / ml) was prepared in DI water. 10 ml of this stock solution was added to 20 ml of the LNP-ID3-NK1 formulation containing 300 mM ammonium sulfate solution and incubated at 40°C for 2 hours. After a 2-hour incubation time, the entire liposome solution containing MTO was dialyzed (12 kDa dialysis membrane, 8-hour dialysis time) to remove free MTO (i.e., not incorporated into the liposomes). After dialysis, an Amicon centrifugal filtration device (cut-off size 10 KDa, 2500 g) was used to concentrate LNP-ID3-NK1 loaded with MTO (LNP-ID3-NK1-MTO).

[0399] The properties of LNP-ID3-NK1-MTO liposomes were determined using a Malvern Zetasizer (Malvern Instrumentation Co., Westborough, MA, USA). Briefly, 2 ml of LNP-ID3-NK1-MTO liposomes (the concentration of liposomes was 0.5 - 1 mg / ml) were placed in a four-sided transparent plastic cuvette and directly analyzed at 25°C. The results shown in Figure 17 indicate that the Zav size of the nanoparticles was approximately 90 nm and the PDI was approximately 0.084.

[0400] Furthermore, the zeta potential of LNP-ID3-NK1-MTO liposomes in an aqueous dispersion was determined using a Malvern zeta seizer Instrument (Malvern Instrumentation Co, Westborough, MA, USA). Briefly, approximately 1 ml of liposomes (concentration approximately 2 mg / ml in 20 mM NaCl) was placed in a disposable capillary zeta potential cell available for the Zetasizer. The measurement was performed at 25°C. The results indicate that the zeta potential determination of LNP-ID3-NK1-MTO was approximately -11.6 mV (Figure 18).

[0401] Example 11: Determination of encapsulation and loading efficiency of LNP-ID3-NK1-MTO liposomes. In another experiment, the encapsulation and loading efficiency of LNP-ID3-NK1-MTO liposomes were determined by disrupting the liposomes and measuring MTO as follows using ultraviolet-visible (UV-vis) spectroscopy. Briefly, after separating the free MTO by dialysis, the liposomes were disrupted with dimethyl sulfoxide. The capture efficiency was determined using the following formula. Encapsulation efficiency (%) = [MTO] f / [MTO] t × 100 where [MTO] f is the concentration of MTO encapsulated in LNP-ID3-NK1, and [MTO] tis the total concentration of MTO (meaning the total amount of MTO initially added: loaded + free). Subsequently, the MTO concentration was determined using a Nanodrop 2000C UV-vis spectrophotometer. The capture efficiency of MTO by LNP-ID3-NK1-MTO was found to be approximately 94%. Furthermore, the loading efficiency of MTO in LNP-ID3-NK1-MTO was determined using the following equation. Loading efficiency (%) = [MTO] fwt / [LNP-ID3-NK1-MTO] fwt × 100 [MTO] fwt is the total weight of MTO encapsulated in MTO, and [LNP-ID3-NK1-MTO] fwt is the total weight of LNP-ID3-NK1 (LNP-ID3-NK1-MTO) encapsulating MTO. The loading efficiency of MTO in LNP-ID3-NK1-MTO was found to be approximately 4.1% w / w.

[0402] Example 12: Tumor inhibition of LNP-ID3 combined with other liposomes using B16F10 cells in vivo The evaluation of LNP-ID3 combined with various liposomes of the present disclosure was performed using the following protocol. Briefly, mouse melanoma cancer B16F10 cells (cells (0.2 × 10 6was inoculated subcutaneously into the right posterior abdominal region of C57BL / 6 mice. Animals were treated twice a week by intravenous injection with vehicle control, 3 mg / kg of LNP-MTO (liposomal mitoxantrone dihydrochloride), a combination of 3 mg / kg of liposomal LNP-ID3 and LNP-TB4 (TGF-β inhibitor-stearic acid), a combination of 3 mg / kg of LNP-ID3 and LNP-TR6 (liposomal TLR1 / 2 agonist-CHEMS), and a combination of 3 mg / kg of LNP-MTO, LNP-ID3 and LNP-TB4. Tumor volume was measured three times in two dimensions using calipers and the volume was calculated using the formula: V = (L × W × W) × 0.5, where V is the tumor volume, L is the length of the tumor (the longest tumor dimension), and W is the width of the tumor (the longest tumor dimension perpendicular to L). Tumor growth inhibition (TGI) was calculated based on tumor size data on day 16.

[0403] The results showed that treatment with LNP-MTO as a single agent exhibited anti-tumor activity and the TGI was calculated to be 32.56% when compared to the vehicle group (p < 0.05). Furthermore, the combination treatment of LNP-TB4 + LNP-MTO + LNP-ID3 had significant anti-tumor activity compared to the vehicle group and the TGI was 32.86% (all p < 0.05). (Figure 19).

[0404] Example 13: Tumor inhibition of LNP-ID3 combined with LNP-AR5 using B16F10 cells in vivo In another experiment, the following protocol was used to evaluate LNP-ID3 in combination with LNP-AR5. Briefly, mouse melanoma cancer B16F10 cells (cells (0.2 × 10 6([[ID=]]) was inoculated subcutaneously into the right posterior abdominal region of C57BL / 6 mice. Animals were treated twice a week by intravenous injection with vehicle control, 3 mg / kg of LNP-AR5, and a combination of 3 mg / kg of LNP-AR5 and LNP-ID3. Tumor volume was measured three times in two dimensions using calipers, and the volume was calculated using the formula: V = (L × W × W) × 0.5, where V is the tumor volume, L is the length of the tumor (the longest tumor dimension), and W is the width of the tumor (the longest tumor dimension perpendicular to L). Tumor growth inhibition (TGI) was calculated based on the tumor size data on day 15.

[0405] The results showed that the combined treatment of LNP-AR5 + LNP-ID3 had significant mild antitumor activity when compared to the vehicle group, with a TGI of 35.89% (all p < 0.05). (Figure 20).

[0406] Example 14: Tumor inhibition of LNP-ID3-NK1 using B16F10 cells in vivo. In another experiment, the following protocol was used to evaluate LNP-ID3-NK1. Briefly, mouse melanoma cancer B16F10 cells (cells (0.2 × 10 6 ) were inoculated subcutaneously into the right posterior abdominal region of C57BL / 6 mice. Animals were treated twice a week by intravenous injection with vehicle control, 2 mg / kg of LNP-MTO (liposomal mitoxantrone dihydrochloride), 3 / 0.25 mg / kg of LNP-ID3-NK1 (in liposomal form at a ratio of 1:12), a combination of 3 mg / kg of LNP-MTO and 3 / 0.25 mg / kg of LNP-ID3-NK1, and a combination of 3 / 0.25 mg / kg of LNP-ID3-NK1 and 5 mg / kg of PD3 (liposomal PD-1 receptor inhibitor-cholesterol). Tumor volume was measured three times in two dimensions using calipers, and the volume was calculated using the formula: V = (L × W × W) × 0.5, where V is the tumor volume, L is the length of the tumor (the longest tumor dimension), and W is the width of the tumor (the longest tumor dimension perpendicular to L). Tumor growth inhibition (TGI) was calculated based on the tumor size data on day 18.

[0407] The results showed that treatment with LNP-ID3-NK1, LNP-MTO + LNP-ID3-NK1, and LNP-ID3-NK1 + LNP-PD3 caused significant tumor growth inhibition compared to the vehicle group, and the TGI was calculated to be 42.71% - 54.31% respectively (p < 0.05). (Figure 21).

[0408] Example 15: Tumor inhibition of LNP-ID3 and LNP-ID3-NK1 using CT26 cells in vivo. In another experiment, LNP-ID3 and LNP-ID3-NK1 were evaluated using the following protocol. Briefly, mouse colon cancer tumor CT26 cells (0.1×106 cells) were subcutaneously inoculated into the right posterior abdominal region of Balb / C mice. The animals were treated twice a week by intravenous injection with vehicle control, 3 mg / kg of LNP-DOX (liposomal doxorubicin), 3 mg / kg of LNP-ID3, 0.25 mg / kg of LNP-NK1, a combination of 3 mg / kg of LNP-DOX and LNP-ID3, 3 / 0.25 mg / kg of LNP-ID3-NK1 (1:12 ratio), and a combination of 3 mg / kg of LNP-DOX and 3 / 0.25 mg / kg of LNP-ID3-NK1. The tumor volume was measured three times in two dimensions using calipers, and the volume was calculated using the formula: V = (L × W × W) × 0.5, where V is the tumor volume, L is the tumor length (the longest tumor dimension), and W is the tumor width (the longest tumor dimension perpendicular to L). The study was terminated when the average tumor size in the vehicle-treated group reached more than 2500 mm3 (on the 27th day after tumor inoculation). Tumor growth inhibition (TGI) was calculated based on the tumor size data on the 27th day.

[0409] The results showed that treatment with LNP-DOX, LNP-DOX + LNP-ID3, and LNP-DOX + LNP-ID3-NK1 caused significant TGI compared to the vehicle group (p < 0.01). The TGI was recorded as 91%, 80%, and 89.2% respectively. (Figure 22).

[0410] Example 16: Tumor inhibition of LNP-ID3 using CT26 cells in vivo. In another experiment, the evaluation of LNP-ID3 was carried out using the following protocol. Mouse colorectal carcinoma CT26 cells (cells (0.1×10 6 )) were inoculated subcutaneously into the right posterior abdominal region of Balb / C mice. Animals were treated twice a week by intravenous injection with vehicle control, 3 mg / kg of LNP-DOX (liposomal doxorubicin), 3 mg / kg of LNP-ID3, a combination of 3 mg / kg of LNP-DOX and LNP-ID3, a combination of 3 mg / kg of LNP-ID3 and 0.25 mg / kg of LNP-NK1, a combination of 3 mg / kg of LNP-ID3 and LNP-TR6, a combination of 3 mg / kg of LNP-ID3 and 4 mg / kg of LNP-TR5, and 3 mg / kg of LNP-ID3 and 2 mg / kg of LNP-TR8 (liposomal 3D-6-acyl-PHAD). Tumor volume was measured three times in two dimensions using calipers, and the volume was calculated using the formula: V=(L×W×W)×0.5, where V is the tumor volume, L is the length of the tumor (the longest tumor dimension), and W is the width of the tumor (the longest tumor dimension perpendicular to L).

[0411] The results show that LNP-DOX alone or in combination with LNP-ID3 conferred a therapeutic benefit by significantly prolonging the survival of CT26 tumor-bearing mice. (Figure 23).

[0412] Example 17: Tumor inhibition of LNP-ID3 using MC-38 cells in vivo. In another experiment, further evaluation of LNP-ID3 was carried out using the following protocol. Mouse colorectal adenocarcinoma MC-38 cells (cells (1×10 6was inoculated subcutaneously into the right posterior abdominal region of C57BL / 6 mice. Vehicle control, combination, 4 mg / kg of LNP-DOX (liposomal doxorubicin), 10 mg / kg of anti-PD1 antibody, combination of 4 mg / kg of LNP-ID3 and LNP-AR5, combination of 4 mg / kg of LNP-DOX + LNP-ID3 + LNP-AR5, and for the first dose, a combination of 3.5 mg / kg of LNP-ID3, 3.5 mg / kg of LNP-AR5 and 4 mg / kg of LNP-TR5, and for the remaining doses at 2 mg / kg, animals were treated twice a week by intravenous injection. Animals were administered LNP-DOX only twice and then switched to vehicle control. Tumor volume was measured three times in two dimensions using calipers and the volume was calculated using the formula: V = (L × W × W) × 0.5, where V is the tumor volume, L is the length of the tumor (the longest tumor dimension), and W is the width of the tumor (the longest tumor dimension perpendicular to L). Tumor growth inhibition (TGI) was calculated based on the tumor size data on day 19.

[0413] The results show that treatment with LNP-DOX, LNP-DOX + LNP-ID3 + LNP-AR5, and LNP-ID3 + LNP-AR5 showed substantial TGI when compared to the vehicle group (p < 0.05). TGI was recorded at 74.39%, 90.40% and 73.77% respectively. (Figure 24).

[0414] Example 18: Tumor inhibition of LNP-ID3 combined with LNP-AR5 using H22 cells in vivo In another experiment, the following protocol was used to evaluate LNP-ID3 combined with LNP-AR5. Briefly, mouse hepatocellular carcinoma H22 cells (cells (2 × 10 6) was inoculated subcutaneously into the right posterior abdominal region of Balb / C mice. Animals were treated twice a week by intravenous injection with vehicle control, and a combination of 4 mg / kg of LNP-ID3 and LNP-AR5. Tumor volume was measured three times in two dimensions using calipers, and the volume was calculated using the formula: V = (L × W × W) × 0.5, where V is the tumor volume, L is the length of the tumor (the longest tumor dimension), and W is the width of the tumor (the longest tumor dimension perpendicular to L). Tumor growth inhibition (TGI) was calculated based on the tumor size data on day 14. The results indicate that LNP-ID3 + LNP-AR5 caused TGI when compared to the vehicle. TGI was recorded at 43.28%. (Figure 25).

[0415] Example 19: Measurement of IDO-1 activity. IDO-1 activity was indirectly measured using kynurenine levels using the following protocol. Briefly, hIDO1-HEK293 recombinant cells were seeded overnight. The next day, the cells were left untreated or treated with 10 uM of ID3, ID3-SA or LNP-ID3-SA for 24 hours. IDO-1 activity was measured by indirect measurement of kynurenine levels by analyzing the absorption at 480 nm using the IDO1 Cellular Activity QuickDetect™ kit (PBSBioscience, San Diego, CA).

[0416] The results indicate that untreated cells showed high levels of IDO-1 activity and that treatment with ID3, ID3-SA and LNP-ID3-SA inhibited IDO-1 activity. These results confirm the activity of ID3 in prodrug and liposomal form. (Figure 26).

[0417] Example 20: Synthesis and characterization of SLNP-AR5-TR5-ID3 solid-lipid nanoparticles In another experiment, Moliwol 488 (polyvinyl alcohol) was used as an emulsifier, and solid-lipid nanoparticles (referred to as "SLNP") containing ID3-AR5-TR5 (SLNP-ID3-AR5-TR5) were prepared using the following protocol. Briefly, lipid stock solutions of HSPC, CHOL, DSPE-PEG, and TR5 were separately prepared in ethanol (20 mg / ml). The ID3 prodrug stock solution was prepared in acetonitrile (20 mg / ml). A lipid mixture of HSPC, CHOL, AR5, TR5, ID3, and DSPE-PEG in a molar ratio of 51.6:27:7:2.4:7:5 was mixed together and then diluted with ethanol to obtain a lipid concentration of 5 mg / ml. This lipid mixture was heated at 50 °C using a heating block attachment in a microfluidizer. Similarly, the aqueous phase containing 2% Moilwol 488 solution was preheated at 50 °C before passing through a microfluidics cartridge (Precision NanoSystems, Inc.) at a flow rate of 4:1 (aqueous phase: organic phase, lipid mixture). The solvent was removed using a dialysis membrane (Sigma Aldrich) with a cut-off of approximately 12 kDa size against DI water for at least 24 hours. To maximize solvent removal, the dialysis water was changed at least 5 times during the 24-hour period. After removing the solvent, the SLNP was passed through a 0.2 micron filter membrane (cellulose acetate). The SLNP-AR5-TR5-ID3 was concentrated using an Amicon centrifugal filtration device (cut-off size 10 kDa, 3000 g).

[0418] The properties of the SLNP-AR5-ID3-TR5 solid-lipid nanoparticles were determined using a Malvern Zetasizer (Malvern Instrumentation Co., Westborough, MA, USA). Briefly, 2 ml of SLNP-AR5-ID3-TR5 (at a concentration of 1 mg / ml) was placed in a four-sided transparent plastic cuvette and analyzed directly at 25 °C. The results shown in Figure 27 indicate that the Zav size of the nanoparticles was approximately 106 nm and the PDI was approximately 0.171.

[0419] Furthermore, the zeta potential of SLNP-AR5-ID3-TR5 solid-lipid nanoparticles in the aqueous dispersion was determined using a Malvern zeta seizer Instrument (Malvern Instrumentation Co, Westborough, MA, USA). Briefly, approximately 1 ml of the nanoparticle formulation (at a concentration of about 3 mg / ml in DI water) was placed into a disposable capillary zeta potential cell available for the Zetasizer. Measurements were taken at 25 °C. The results indicate that the zeta potential determination of SLNP-AR5-ID3-TR5 was about 9.87 mV (Figure 28).

[0420] Example 21: Human Clinical Trials for the Treatment of Human Cancer by the Use of Formulated and / or Co-formulated Liposomes Containing an IDO Prodrug The formulated and / or co-formulated liposomes containing an IDO prodrug, which specifically accumulate in tumor cells and are used in the treatment of certain tumors and other immune disorders and / or other diseases, are used according to the present invention. For each of these indications, two clinical approaches are successfully pursued.

[0421] I.) Adjuvant Therapy: In adjuvant therapy, patients are treated with formulated and / or co-formulated liposomes containing an IDO prodrug in combination with a chemotherapeutic agent or a pharmaceutical or biopharmaceutical agent or a combination thereof. The primary cancer target is treated under a standard protocol with the addition of formulated and / or co-formulated liposomes containing an IDO prodrug. The protocol design addresses the efficacy, including but not limited to examples such as reduction in tumor mass of primary or metastatic lesions, prolongation of progression-free survival, overall survival, improvement in the patient's health status, disease stabilization, and the ability to reduce the normal dosages of standard chemotherapy and other biological agents. These dosage reductions enable additional and / or long-term treatment by reducing the dose-related toxicity of the chemotherapeutic or biological agent.

[0422] II.) Monotherapy: Regarding the use of formulated and / or co-formulated liposomes containing an IDO prodrug in the monotherapy of tumors, the formulated and / or co-formulated liposomes containing an IDO prodrug are administered to patients without a chemotherapeutic agent or a pharmaceutical or biological factor. In one embodiment, the monotherapy is clinically implemented in patients with advanced cancer having a wide range of metastatic diseases. The protocol design addresses the effectiveness evaluated by examples including, but not limited to, reduction in tumor volume of primary or metastatic lesions, extension of progression-free survival, overall survival, improvement in the patient's health status, disease stabilization, and the ability to reduce the normal doses of standard chemotherapy and other biological agents. Dosage

[0423] The dosing regimen can be adjusted to provide the optimal desired response. For example, a single formulated and / or co-formulated liposome containing an IDO prodrug can be administered, or several divided doses can be administered over time, and the dose can also be proportionally decreased or increased as indicated by the urgency of the treatment situation. As used herein, "dosage unit form" refers to physically discrete units suitable as a unit dosage for the mammalian subject to be treated, each unit containing a predetermined quantity of the active compound calculated to produce the desired therapeutic effect in association with the required pharmaceutical carrier. The specification of the dosage unit form of the present invention is determined by and directly depends on (a) the unique characteristics of the formulated and / or co-formulated liposomes containing an IDO prodrug, (b) the individual mechanisms of the co-administered compounds, if any, (c) the specific therapeutic or prophylactic effect to be achieved, and (d) the limitations inherent in the art of compounding such compounds for the treatment of sensitivity in an individual. Clinical Development Plan (CDP)

[0424] CDP develops this in accordance with a treatment using formulated and / or co-formulated liposomes containing an IDO prodrug, in relation to adjuvant therapy or monotherapy. The trials first demonstrate safety and then confirm efficacy in repeated dosing. The trials are non-blinded and compare standard chemotherapy and / or current standard treatments, with the addition of formulated and / or co-formulated liposomes containing an IDO prodrug. As will be understood, one non-limiting criterion that can be utilized in relation to patient enrollment is the expression of IDO-1 in tumors determined by standard detection methods known in the art.

[0425] It is contemplated that any of the formulated and / or co-formulated liposomes, or embodiments disclosed herein, may have satisfactory pharmacological profiles and promising biopharmaceutical properties, such as toxicological profiles, metabolism and pharmacokinetic properties, solubility and permeability. It will be understood that the determination of appropriate biopharmaceutical properties, such as cytotoxicity in cells to determine potential toxicity or inhibition of certain targets or channels, is within the knowledge of those skilled in the art.

[0426] The present invention should not be limited in scope by the embodiments disclosed herein, which are intended as single illustrations of individual aspects of the present invention, and any functionally equivalent ones are within the scope of the present invention. In addition to what is described herein, various modifications to the models, methods, and life cycle methods of the present invention will be apparent to those skilled in the art from the foregoing description and teachings and are likewise intended to be within the scope of the present invention. Such modifications or other embodiments can be implemented without departing from the true scope and spirit of the present invention.

[0427] [Table 1] [Table 2] [Table 3] The present invention provides, for example, the following items. (Item 1) An IDO prodrug composition, wherein the IDO prodrug composition comprises (i) a drug moiety, (ii) a lipid moiety, (iii) a linking unit ("LU") and the drug moiety contains an IDO antagonist, and the LU conjugates the drug moiety with the lipid moiety, an IDO prodrug composition. (Item 2) The IDO prodrug according to Item 1, wherein the drug moiety contains a chemical structure represented as ID3. (Item 3) The IDO prodrug according to Item 1, wherein the lipid moiety contains CHEMS. (Item 4) The IDO prodrug according to Item 1, wherein the lipid moiety contains stearic acid. (Item 5) A liposome (LPN) containing an IDO prodrug, wherein the liposome releases an active IDO inhibitor after cleavage of the LU, a liposome (LPN). (Item 6) The liposome according to Item 5, wherein the IDO prodrug contains ID3. (Item 7) The liposome according to Item 5, wherein the IDO prodrug contains ID3 and further contains TR3. (Item 8) The liposome according to Item 5, wherein the IDO prodrug contains ID3 and further contains TR5. (Item 9) The liposome according to Item 8, wherein TR5 contains telratrimod. (Item 10) The liposome according to Item 5, wherein the IDO prodrug contains ID3 and further contains AR5. (Item 11) The liposome according to Item 10, wherein AR5 contains an A2a receptor inhibitor. (Item 12) The liposome is further co-formulated with an immunomodulatory agent, and the immunomodulatory agent is selected from the group consisting of immunogenic cell death-inducing chemotherapeutic agents, toll-like receptor agonists, STING agonists, CTLA-4 inhibitors, PD-1 / PD-L1 inhibitors, and / or prodrugs thereof. The liposome according to item 5. (Item 13) The liposome is further co-formulated with an ICD-inducing chemotherapeutic agent, and the ICD-inducing chemotherapeutic agent is selected from the group consisting of DOX, MTO, OXA, CP, bortezomib, carfilzomib, or paclitaxel. The liposome according to item 5. (Item 14) The liposome according to item 5, further comprising DOX. (Item 15) The liposome according to item 5, further comprising MTO. (Item 16) The liposome according to item 13, further comprising DOX. (Item 17) The liposome according to item 13, further comprising MTO. (Item 18) A method of treating a subject suffering from or diagnosed with cancer, (i) administering to a subject in need of such treatment an effective amount of a liposome, wherein the liposome comprises an IDO prodrug, and (ii) its pharmaceutically acceptable salts, A method comprising. (Item 19) The method according to item 18, wherein the IDO prodrug comprises ID3. (Item 20) The method according to item 18, wherein the cancer is colorectal cancer.

Claims

1. Use of a nanocarrier in the manufacture of a medicament for treating a subject suffering from or diagnosed with cancer, wherein the nanocarrier comprises an IDO prodrug, the IDO prodrug having the following chemical structure: 【Chemistry 2】 or a pharmaceutically acceptable salt thereof.

2. The use described in claim 1, wherein the nanocarrier is a liposome (LPN).

3. The use described in claim 1, wherein the nanocarrier is a solid lipid nanoparticle (SLNP).

4. The use described in claim 2, wherein the liposome further contains α-galactosylceramide (α-GalCer).

5. The use described in claim 2, wherein the liposome further contains telluritol.

6. The liposome further comprises an A2a receptor inhibitor, wherein the A2a receptor inhibitor has the following chemical structure: 【Chemistry 19】 The use according to claim 2, consisting of:

7. The use described in claim 6, wherein the liposomes have a Zav size of 91 nm and a PDI of 0.

212.

8. The use described in claim 5, wherein the liposomes have a Zav size of 96 nm and a PDI of 0.

117.

9. The use described in claim 4, wherein the liposomes have a Zav size of 80 nm and a PDI of 0.

189.

10. The solid lipid nanoparticles further comprising an A2a receptor inhibitor, wherein the A2a receptor inhibitor has the following chemical structure: 【Chemistry 19】 The use according to claim 3, consisting of:

11. The use described in claim 3, wherein the solid lipid nanoparticles further comprise telluritolimod.

12. The use of claim 3, wherein the solid lipid nanoparticles further comprise α-galactosylceramide (α-GalCer).

13. The use of claim 10, wherein the solid lipid nanoparticles have a Zav size of 106 nm and the solid lipid nanoparticles have a PDI of 0.

171.

14. The use described in claim 2, wherein the liposome further contains doxorubicin (DOX).

15. The use described in claim 2, wherein the liposome further contains mitoxantrone (MTO).

16. The use of claim 3, wherein the solid lipid nanoparticles further comprise doxorubicin (DOX).

17. The use of claim 3, wherein the solid lipid nanoparticles further comprise mitoxantrone (MTO).

18. The use described in claim 1, wherein the cancer is colorectal cancer.

19. The use described in claim 1, wherein the cancer is colon melanoma.

20. The use described in claim 1, wherein the cancer is liver cancer.

21. A composition comprising a nanocarrier for treating a subject suffering from or diagnosed with cancer, wherein the nanocarrier comprises an IDO prodrug, the IDO prodrug having the following chemical structure: 【Chemistry 2】 or a pharmaceutically acceptable salt thereof.

22. The composition described in claim 21, wherein the nanocarrier is a solid lipid nanoparticle (SLNP).

23. The composition described in claim 21, wherein the nanocarrier is a liposome, and the liposome further contains α-galactosylceramide (α-GalCer).

24. The composition described in claim 21, wherein the nanocarrier is a liposome, and the liposome further contains telluritol.

25. The nanocarrier according to claim 25, wherein the nanocarrier is a liposome, the liposome further comprises an A2a receptor inhibitor, and the A2a receptor inhibitor has the following chemical structure: 【Chemistry 19】 22. The composition of claim 21, consisting of:

26. The composition described in claim 25, wherein the nanocarrier is a liposome, the liposome has a Zav size of 91 nm, and the liposome has a PDI of 0.

212.

27. The composition described in claim 24, wherein the nanocarrier is a liposome, the liposome has a Zav size of 96 nm, and the liposome has a PDI of 0.

117.

28. The composition described in claim 23, wherein the nanocarrier is a liposome, the liposome has a Zav size of 80 nm, and the liposome has a PDI of 0.

189.

29. The solid lipid nanoparticles further comprising an A2a receptor inhibitor, wherein the A2a receptor inhibitor has the following chemical structure: 【Chemistry 19】 23. The composition of claim 22, consisting of:

30. The composition of claim 22, wherein the solid lipid nanoparticles further comprise telluritol.

31. The composition of claim 22, wherein the solid lipid nanoparticles further comprise α-galactosylceramide (α-GalCer).

32. The composition of claim 29, wherein the solid lipid nanoparticles have a Zav size of 106 nm and the solid lipid nanoparticles have a PDI of 0.

171.

33. The composition described in claim 21, wherein the nanocarrier is a liposome, and the liposome further contains doxorubicin (DOX).

34. The composition described in claim 21, wherein the nanocarrier is a liposome, and the liposome further contains mitoxantrone (MTO).

35. The composition of claim 22, wherein the solid lipid nanoparticles further comprise doxorubicin (DOX).

36. The composition of claim 22, wherein the solid lipid nanoparticles further comprise mitoxantrone (MTO).

37. The composition described in claim 21, wherein the cancer is colorectal cancer.

38. The composition described in claim 21, wherein the cancer is colon melanoma.

39. The composition described in claim 21, wherein the cancer is liver cancer.

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