Compounds for modulating trained immunity and methods of use thereof - Patents.com

Abstract

Provided herein are compounds of formula (I), as well as compositions comprising compounds of formula (I) and uses thereof. [Formula 1] TIFF2024511067000110.tif2552

Description

[Technical field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application claims priority to U.S. Provisional Patent Application No. 63 / 163,428, filed March 19, 2021, the disclosure of which is incorporated by reference in its entirety for all purposes.

[0002] Description of electronically submitted text files The contents of the text file submitted electronically herewith are incorporated herein by reference in their entirety: Copy of Sequence Listing in Computer Readable Format (Filename: TRAI_005_00US_SeqList_ST25.txt, Recorded on: March 17, 2021, File Size: 105 KB). [Background technology]

[0003] The immune system plays a key role in the pathophysiology of major diseases such as atherosclerosis, diabetes, and cancer. However, most of the immunotherapeutic strategies currently developed focus either on effector molecules such as cytokines, or on T lymphocytes, cells from the adaptive immune system (Mulder et al. Nat. Rev. Drug Discov. 2019, 18(7), 553-566; Pardoll et al. Nat Immunol., 2012, 13, 1129-1132). In autoimmune and autoinflammatory diseases, bioactive cytokines can be neutralized by anti-cytokine therapy, but the most commonly used immunotherapies for cancer patients include the application of checkpoint inhibitors. Although it was long thought that the innate immune system had no memory, recent studies have shown that innate immune cells undergo metabolic and epigenetic rewiring to adjust their functional programs in a process termed "trained immunity" that is involved in exerting antitumor effects (Buffen et al., 2014, PLoS Pathog. 10, e1004485; Netea et al., J. Leukoc Biol. 2017, 102, 1323-1332).

[0004] A series of pattern recognition receptors (PRRs), including TLRs, NOD2, dectin-1 and inflammasomes, can be engaged to promote trained immunity. In addition, in vitro studies have demonstrated that BCG, and several other PAMPs and DAMPs, including peptidoglycan and β-glucan, can be therapeutically utilized as trained immunity stimulators. However, the in vivo therapeutic utilization of molecules that regulate trained immunity has been hindered by toxicity, immune-related adverse effects, and poor bioavailability targeting relevant myeloid cells and their progenitors.

[0005] There is a need for therapeutic agents and compositions thereof that involve the innate immune system and modulate trained immunity to treat cancer and other diseases and conditions caused by defective trained immunity. Summary of the Invention

[0006] In embodiments, provided herein is a compound having formula (I):

[0007] [ka] or a pharma- ceutically acceptable salt thereof. (In the formula, R 1 is -H or -C(O)-R X and; R 2 and R 3 are each independently selected from the group consisting of -H, alkyl, alkylene-aryl, -C(O)-alkyl, and -C(O)-aryl; R 4 , R 5 , and R 5’ are each alkyl; R 6 and R 11 are each independently -H or alkyl; R 7 is C 9~30 Fatty acid chain, -YN(R 11 )-C(O)-O-alkylene-C(H)(OR 8 )-Alkylene-OR 9 , -C(R 10 )(C(O)NH2)-alkylene-N(R 11 )-C(O)-C 16~30 Fatty acid chain, -(CR 10 R 10 )2-OP(O)(OH)-O-alkylene-C(R 10 )(OR Z )-Alkylene-OR Z’ or -Y-triazolyl-L; R Z and R Z’ are each independently C 8~30 Fatty acids or -C(O)-C 16~30 is a fatty acid chain; Y is alkylene; L is a fatty acid chain, -alkylene-C(O)-W, -alkylene-OC(O)-W, -alkylene-N-(alkylene-C(O)-NR 11 -Alkylene-NR 11 -C(O)-W)2, and -alkylene-N-(alkylene-C(O)-W)2; W is a fatty acid chain, -O-alkylene-C(H)(OR 8 )-Alkylene-OR 9 , a phospholipid, or a sterol; R 8 and R 9 are each independently R X or -C(O)-R X and; R 10 , R 22 , R 33 , R 33’ , R 44 , R 44’ , R 55 , and R 55’ are each independently H or R A and; R X is a fatty acid chain; Each of the foregoing alkyl, alkylene, alkylene-aryl, aryl, and triazolyl may be selected from one or more R A Optionally replaced by R A is hydrogen, halo, alkoxy, haloalkoxy, cyano, hydroxyl, -N(R C )(R D ), -C(O)N(R C )(R D ), -N(R C )C(O)R B , -OC(O)NR C R D , -NR C C(O)OR B , -OC(O)R B , -C(O)OR B , -C(O)R B , -CO2H, -NO2, -SH, S(O) X R B(wherein X is 0, 1, or 2), aryl, arylalkyl, heteroaryl, heteroarylalkyl, cycloalkyl, heterocycloalkyl, and R B independently selected for each occurrence from the group consisting of: R C and R D is hydrogen, alkyl, haloalkyl-C(O)R B , and -C(O)OR B or R C and R D together with the nitrogen to which they are attached, R A forming an optionally substituted heterocycle; R B is alkyl, alkenyl, or alkynyl optionally substituted with one or more fluoro; R 7 C 9~30 If it is a fatty acid chain, R 2 is -H) It is.

[0008] In an embodiment, the compound of formula (I) has the formula (IA):

[0009] [ka] or a pharma- ceutically acceptable salt thereof. (In the formula, R 1 , R 2 , R 3 , R 4 , R 5 , R 55 , R 5’ , R 55’ , R 33 , R 33’ , R 44 , R 44’ , R 6 , and R 7 is defined herein) It is.

[0010] In an embodiment, the compound of formula (I) has the formula (IB):

[0011] [ka] It is. (In the formula, R 2 , R 3 , R 4 , R 6 , and R 7 is defined herein)

[0012] In an embodiment, the compound of formula (I) has formula (II):

[0013] [ka] or a pharma- ceutically acceptable salt thereof. (In the formula, R 2 , R 3 , R 4 , R 5 , R 5’ , R 6 , Y, X 1 , X 2 , R A , L is defined herein. It is.

[0014] In an embodiment, the compound of formula (II) is

[0015] [ka] or a pharma- ceutically acceptable salt thereof (In the formula, R 2 , R 3 , R 4 , R 5 , R 5’ , R 6 , Y, X 1 , X 2 and L is defined herein. It is.

[0016] In an embodiment, the compound of formula (I) has the formula (IIA):

[0017] [ka] or a pharma- ceutically acceptable salt thereof. (In the formula, R 2 , R 3 , R 4 , R 5 , R 5’ , R 6 , Y, X 1 , X 2 , L, and R A is defined herein) It is.

[0018] In an embodiment, the compound of formula (IIA) is

[0019] [ka] It is. (In the formula, R 2 , R 3 , R 4 , R 5 , R 5’ , R 6 , Y, X 1 , X 2 and L is defined herein.

[0020] In embodiments, the present disclosure provides a nanobiological composition comprising high density lipoprotein (HDL)-derived nanoparticles, the nanoparticles having formula (I);

[0021] [ka] or a pharma- ceutically acceptable salt thereof. (In the formula, R 1 is -H or -C(O)-R X and; R 2 and R 3 are each independently selected from the group consisting of -H, alkyl, alkylene-aryl, -C(O)-alkyl, and -C(O)-aryl; R 4 , R 5, and R 5’ are each alkyl; R 6 and R 11 are each independently -H or alkyl; R 7 is the fatty acid chain, -YN(R 6 )-C(O)-O-alkylene-C(H)(OR 8 )-Alkylene-OR 9 , -YN(R 6 )-C(O)-R X , -YOP(O)(OH)-O-alkylene-C(H)(OR 8 )-Alkylene-OR 9 or -Y-triazolyl-L; Y is alkylene; L is a fatty acid chain, -alkylene-C(O)-W, -alkylene-OC(O)-W, -alkylene-N-(alkylene-C(O)-NR 11 -Alkylene-NR 11 -C(O)-W)2, and -alkylene-N-(alkylene-C(O)-W)2; W is a fatty acid chain, -O-alkylene-C(H)(OR 8 )-Alkylene-OR 9 , a phospholipid, or a sterol; R 8 and R 9 are each independently R X or -C(O)-R X and; R 10 , R 22 , R 33 , R 33’ , R 44 , R 44’ , R 55 , and R 55’ are each independently H or R A and; R X is a fatty acid chain; Each of the foregoing alkyl, alkylene, alkylene-aryl, aryl, and triazolyl may be selected from one or more R A Optionally replaced by RA is hydrogen, halo, alkoxy, haloalkoxy, cyano, hydroxyl, -N(R C )(R D ), -C(O)N(R C )(R D ), -N(R C )C(O)R B , -OC(O)NR C R D , -NR C C(O)OR B , -OC(O)R B , -C(O)OR B , -C(O)R B , -CO2H, -NO2, -SH, S(O) X R B (wherein X is 0, 1, or 2), aryl, heteroaryl, heteroarylalkyl, cycloalkyl, heterocycloalkyl, and R B independently selected for each occurrence from the group consisting of: R C and R D is hydrogen, alkyl, haloalkyl-C(O)R B , and -C(O)OR B or R C and R D together with the nitrogen to which they are attached, R A forming an optionally substituted heterocycle; R B is alkyl, alkenyl, or alkynyl optionally substituted with one or more fluoro. Includes.

[0022] In embodiments, the present disclosure provides a method for treating a cell proliferation disorder, or sepsis, in a patient in need thereof, comprising administering to the patient a therapeutically effective amount of a compound or nanobiological composition disclosed herein. [Brief description of the drawings]

[0023] [Figure 1]1 shows cryo-TEM photographs of HDL-derived nanoparticles from entries A-F of Table 8 in Example 24. The 50 nm bar applies to all six photographs. [Diagram 2] 2 shows DLS-determined Z-average diameter and PDI values ​​of nanoparticles in Formulations A to F of Formulation 1 in Example 23. [Diagram 3] 2 is a graph showing OD values ​​as a function of test article concentration in the NOD2 activation assay described in Example 25. [Figure 4] AE show tumor growth curves for the study described in Example 26. [Diagram 5] AC show tumor growth curves for the study described in Example 27. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0024] definition For convenience, certain terms used in the specification, examples, and claims are collected here. Unless otherwise defined, all technical and scientific terms used in this disclosure have the same meaning as commonly understood by one of ordinary skill in the art to which this disclosure belongs.

[0025] Throughout this disclosure, various patents, patent applications and publications are referenced. The disclosures of these patents, patent applications and publications are incorporated by reference in their entirety into this disclosure in order to more fully describe the state of the art known to those skilled in the art as of the date of this disclosure. This disclosure shall control in the event of any inconsistency between the cited patents, patent applications and publications and this disclosure.

[0026] The term "about" immediately preceding a numerical value means a range of plus or minus the degree of variation accepted in the art. In an embodiment, unless otherwise indicated in the context of this disclosure or inconsistent with such an interpretation, the term "about" encompasses 10% of the value, e.g., "about 50" means 45-55, "about 25,000" means 22,500-27,500, etc. For example, in a list of numerical values ​​such as "about 49, about 50, about 55, ~", "about 50" means a range that spans less than half the interval between the preceding and following values, e.g., 49.5 or more and less than 52.5. Furthermore, the phrases "less than about a value" or "greater than about a value" should be understood in light of the definition of the term "about" provided herein.

[0027] As used herein, the phrase "pharmacologically acceptable" refers to those compounds, materials, compositions and / or dosage forms which are, within the scope of sound medical judgment, suitable for use in contact with the tissues of human beings and animals without undue toxicity, irritation, allergic response, or other problem or complication, commensurate with a reasonable benefit / risk ratio.

[0028] "Salt" includes compounds that function as bases and inorganic or organic acids to form salts, or compounds that function as acids and inorganic or organic bases to form salts. "Salt" includes derivatives of active agents, which are modified by making their acid or base addition salts. Preferably, the salt is a pharmaceutically acceptable salt. Such salts include, but are not limited to, pharmaceutically acceptable acid addition salts, pharmaceutically acceptable base addition salts, pharmaceutically acceptable metal salts, ammonium salts, and alkylated ammonium salts. Acid addition salts include salts of inorganic and organic acids. Representative examples of suitable inorganic acids include hydrochloric acid, hydrobromic acid, hydroiodic acid, phosphoric acid, sulfuric acid, nitric acid, and the like. Representative examples of suitable organic acids include formic acid, acetic acid, trichloroacetic acid, trifluoroacetic acid, propionic acid, benzoic acid, 3-(4-hydroxybenzoyl)benzoic acid, cinnamic acid, citric acid, fumaric acid, glycolic acid, lactic acid, maleic acid, malic acid, malonic acid, mandelic acid, oxalic acid, picric acid, pyruvic acid, salicylic acid, succinic acid, methanesulfonic acid, ethanesulfonic acid, 2-hydroxyethanesulfonic acid, tartaric acid, ascorbic acid, pamoic acid, bismethylenesalicylic acid, Examples of such salts include ethanedisulfonic acid, gluconic acid, citraconic acid, aspartic acid, stearic acid, palmitic acid, EDTA, glycolic acid, p-aminobenzoic acid, glutamic acid, benzenesulfonic acid, 4-chlorobenzenesulfonic acid, 2-naphthalenesulfonic acid, camphorsulfonic acid, p-toluenesulfonic acid, sulfates, nitrates, phosphates, perchlorates, borates, acetates, benzoates, hydroxynaphthoates, glycerophosphates, and ketoglutarates.Examples of base addition salts include, but are not limited to, ethylenediamine, N-methylglucamine, lysine, arginine, ornithine, choline, N,N'-dibenzylethylenediamine, chloroprocaine, diethanolamine, procaine, N-benzylphenethylamine, diethylamine, piperazine, tris-(hydroxymethyl)-aminomethane, tetramethylammonium hydroxide, triethylamine, dibenzylamine, ephenamine, dehydroabietylamine, N-ethylpiperidine, benzylamine, tetramethylammonium, tetraethylammonium, methylamine, dimethylamine, trimethylamine, ethylamine, basic amino acids such as lysine and arginine dicyclohexylamine, etc. Examples of metal salts include lithium, sodium, potassium, magnesium, calcium salts, etc. Examples of ammonium salts and alkylated ammonium salts include ammonium, methylammonium, dimethylammonium, trimethylammonium, ethylammonium, hydroxyethylammonium, diethylammonium, butylammonium, tetramethylammonium salts, etc. Examples of organic bases include lysine, arginine, guanidine, diethanolamine, choline, etc. Standard methods for preparing pharma- ceutically acceptable salts and their formulations are well known in the art and are disclosed in various references, including, for example, “Remington: The Science and Practice of Pharmacy”, A. Gennaro, ed., 20th edition, Lippincott, Williams & Wilkins, Philadelphia, PA.

[0029] The terms "carrier" or "vehicle" as used interchangeably herein include carriers, excipients, adjuvants, and diluents, or any combination thereof, and refer to a pharma- ceutically acceptable material, composition, or vehicle, such as a liquid or solid filler, diluent, excipient, solvent, or encapsulating material, involved in the carrying or transport of a drug from one organ or part of the body to another. In addition to adjuvants, excipients, and diluents known to those skilled in the art, carriers include nanoparticles of organic and inorganic nature.

[0030] For example, in embodiments, the present disclosure provides nanoparticle carriers (e.g., HDL-derived nanoparticles) as delivery vehicles for active agents (e.g., compounds of formula (I), (IA), (IB), (II), (II-1), (II-2), (IIA), (IIA-1), (IIA-2) or Table 1). In embodiments, the agent is encapsulated within the nanoparticle carrier. In other embodiments, the agent is bound to the surface of the nanoparticle carrier. Association of the agent with the nanoparticle carrier can be achieved by a variety of means, including non-covalent binding, entrapment of the agent within the delivery vehicle, and the like. In embodiments, the association is sufficiently stable such that the agent remains associated with the delivery vehicle until it is delivered to the target site of the subject to be treated.

[0031] As used herein, the term "pharmaceutical combination", "therapeutic combination" or "combination" refers to a single dosage form containing at least two therapeutically active agents, or separate dosage forms containing at least two therapeutically active agents together or separately for use in combination therapy. Administration of the combination therapy includes sequential, simultaneous, or sequential administration in the same or different compositions and / or combinations via the same or different routes. For example, one therapeutically active agent may be formulated in one dosage form and the other therapeutically active agent may be formulated in a single or different dosage form. For example, a first therapeutically active agent may be formulated in a solid oral dosage form and a second therapeutically active agent may be formulated in a solution dosage form for parenteral administration. In embodiments, the combination therapy optionally includes one or more pharma- ceutically acceptable carriers or excipients, non-pharmaceutical active compounds, and / or inactive substances.

[0032] As used herein, the phrase "disorder characterized by cell proliferation" or "condition characterized by cell proliferation" includes, but is not limited to, cancer, benign and malignant tumors. Examples of cancers and tumors include, but are not limited to, cancer or tumor proliferation of the bladder, blood vessels, bone, brain, breast, cervix, breast, colon, endometrium, esophagus, eye, head, kidney, liver, lymph nodes, lung, mouth, cervix, ovary, pancreas, prostate, rectum, colorectum, skin, stomach, testicle, throat, thyroid, urothelium, and uterus.

[0033] The terms "treat", "treating" or "treatment" with respect to a particular disease or disorder include prevention of the disease or disorder and / or reduction, amelioration, remission or arrest of the symptoms and / or pathology of the disease or disorder. In general, the terms used herein refer to amelioration, alleviation, relief, and elimination of symptoms of a disease or condition. The candidate compounds described herein can be present in a formulation or medicament in a therapeutically effective amount, which is an amount that can cause a biological effect such as apoptosis of a particular cell (e.g., cancer cells), reduction in proliferation of a particular cell, or lead to amelioration, alleviation, relief, or elimination of symptoms of a disease or condition such as sepsis. The terms can also refer to a slowing down or halting of cell proliferation (e.g., slowing or stopping tumor growth), or a reduction in the number of proliferating cancer cells (e.g., removing part or all of a tumor).

[0034] As used herein, the term "patient" or "subject" includes all mammals, and more specifically, humans. The methods described herein can be useful for both human therapy and veterinary applications. In one embodiment, the subject is a human.

[0035] As used herein, "prevention" or "preventing" refers to a reduction in the risk of contracting a given disease or disorder, such as preventing the development of at least one clinical symptom of a disease in a subject who may be exposed to or predisposed to the disease, but who has not yet experienced or displayed symptoms of the disease.

[0036] As used herein, "therapeutically effective amount" refers to the amount of a compound or therapeutically active agent that, when administered to a subject for treating a disease or other undesirable medical condition, is sufficient to produce a beneficial effect with respect to the disease or condition. The therapeutically effective amount will vary depending on the type of compound or therapeutically active agent selected, the disease or condition and its severity, as well as the age, weight, etc., of the patient to be treated.

[0037] "Optional" or "optionally" means that the subsequently described event or circumstance may or may not occur, and the description includes examples where the event or circumstance occurs and where it does not. For example, "optionally substituted aryl" includes both "aryl" and "substituted aryl" as defined below. With respect to any group that contains one or more substituents, those of skill in the art will understand that such groups are not intended to introduce substitutions or substitution patterns that are sterically impractical, synthetically impractical, and / or inherently unstable.

[0038] It is further noted that the claims may be drafted to exclude optional elements, and therefore, this statement is intended to serve as a predicate for using exclusive terminology, such as "solely," "only," and the like in connection with the recitation of claim elements or the use of a "negative" limitation.

[0039] When a range of values ​​is listed, it is intended to encompass each value and subrange within that range. For example, C1-C6 alkyl means C1, C2, C3, C4, C5, C6, C7, C8, C9, C10, C11, C12, C13, C14, C15, C16, C17, C18, C19, C20, C21, C22, C23, C24, C25, C26, 1~6 , C 1~5 , C1~4 , C 1~3 , C 1~2 , C 2~6 , C 2~5 , C 2~4 , C 2~3 , C 3~6 , C 3~5 , C 3~4 , C 4~6 , C 4~5 , and C 5~6 Alkyl is intended to be encompassed.

[0040] The term "acyl" as used herein refers to the group RC(O)-, including but not limited to (alkyl)-C(O)-, (alkenyl)-C(O)-, (alkynyl)-C(O)-, (aryl)-C(O)-, (cycloalkyl)-C(O)-, (heteroaryl)-C(O)-, and (heterocyclyl)-C(O)-, where the group is attached to the parent molecular structure via a carbonyl functionality. In embodiments, this refers to the carbonyl carbon of the acyl and the C, which refers to the total number of chain or ring atoms, for example, of the alkyl, alkenyl, alkynyl, aryl, cycloalkyl, or heteroaryl moiety. 1~10 An acyl radical. For example, a C4-acyl has a carbonyl and three other ring or chain atoms.

[0041] An "alkyl" or "alkyl group" is a fully saturated, straight or branched hydrocarbon chain. In embodiments, an alkyl group contains 1 to 30 carbon atoms. In embodiments, an alkyl group has 1 to 12 carbon atoms and is attached to the remainder of the molecule by a single bond. For example, alkyls containing any number of carbon atoms from 1 to 12 are included. Alkyl groups containing up to 12 carbon atoms include C1-C 12 Alkyl, alkyl containing up to 10 carbon atoms is C1-C 10An alkyl group is one having up to 6 carbon atoms, which is C1-C6 alkyl, and one having up to 5 carbon atoms, which is C1-C5 alkyl. C1-C5 alkyl includes C5 alkyl, C4 alkyl, C3 alkyl, C2 alkyl, and C1 alkyl (i.e., methyl). C1-C6 alkyl includes all of the moieties described above for C1-C5 alkyl, but also includes C6 alkyl. C1-C 10 Alkyl includes all of the moieties described above for C1-C5 alkyl and C1-C6 alkyl, but also includes C7, C8, C9 and C 10 Alkyl is also included. Similarly, C1-C 12 Alkyl includes all of the above moieties, except C 11 and C 12 Alkyl is also included. C1~C 12 Non-limiting examples of alkyl include methyl, ethyl, n-propyl, i-propyl, sec-propyl, n-butyl, i-butyl, sec-butyl, t-butyl, n-pentyl, t-amyl, n-hexyl, n-heptyl, n-octyl, n-nonyl, n-decyl, n-undecyl, and n-dodecyl. Unless otherwise stated herein, alkyl groups can be optionally substituted. In embodiments, "alkyl" is a straight chain hydrocarbon. In embodiments, "alkyl" is a branched chain hydrocarbon.

[0042] "Alkylene" or "alkylene chain" refers to a fully saturated, straight or branched divalent hydrocarbon chain. In embodiments, an alkylene group has 1 to 12 carbon atoms. C1-C 12 Non-limiting examples of alkylene include methylene, ethylene, propylene, n-butylene, ethenylene, propenylene, n-butenylene, and the like. The alkylene chain is attached to the rest of the molecule through a single bond and to the group through a single bond. The points of attachment of the alkylene chain to the rest of the molecule and to the group can be through one carbon or any two carbons within the chain. Unless stated otherwise in the specification, the alkylene chain can be optionally substituted.

[0043] "Alkenyl" or "alkenyl group" refers to a straight or branched hydrocarbon chain. In embodiments, an alkenyl group contains 1-30 carbon atoms. In embodiments, an alkenyl group contains 2-12 carbon atoms and has one or more carbon-carbon double bonds, e.g., a straight or branched group of 2-8 carbon atoms, referred to herein as a C2-C8 alkenyl. Each alkenyl group is attached to the remainder of the molecule by a single bond. Alkenyl groups containing any number of carbon atoms from 2 to 12 are included. Alkenyl groups containing up to 12 carbon atoms are C2-C8 alkenyls. 12 Alkenyl, containing up to 10 carbon atoms, is C2-C 10 Alkenyl is an alkenyl group containing up to 6 carbon atoms, C2-C6 alkenyl, and an alkenyl containing up to 5 carbon atoms is C2-C5 alkenyl. C2-C5 alkenyl includes C5 alkenyl, C4 alkenyl, C3 alkenyl, and C2 alkenyl. C2-C6 alkenyl includes all of the moieties listed above for C2-C5 alkenyl, but also includes C6 alkenyl. C2-C 10 Alkenyl includes all of the moieties listed above for C2-C5 alkenyl and C2-C6 alkenyl, but also includes C7, C8, C9 and C 10 Alkenyl is also included. Similarly, C2-C 12 Alkenyl includes all of the above moieties, except that C 11 and C 12 Alkyl is also included. C2~C 12Non-limiting examples of alkenyl include ethenyl (vinyl), 1-propenyl, 2-propenyl (allyl), iso-propenyl, 2-methyl-1-propenyl, 1-butenyl, 2-butenyl, 3-butenyl, 1-pentenyl, 2-pentenyl, 3-pentenyl, 4-pentenyl, 1-hexenyl, 2-hexenyl, 3-hexenyl, 4-hexenyl, 5-hexenyl, 1-heptenyl, 2-heptenyl, 3-heptenyl, 4-heptenyl, 5-heptenyl, 6-heptenyl, 1-octenyl, 2-octenyl, 3-octenyl, 4-octenyl, 5-octenyl, 6-octenyl, 7-octenyl, 1-nonenyl, 2-nonenyl, 3-nonenyl , 4-nonenyl, 5-nonenyl, 6-nonenyl, 7-nonenyl, 8-nonenyl, 1-decenyl, 2-decenyl, 3-decenyl, 4-decenyl, 5-decenyl, 6-decenyl, 7-decenyl, 8-decenyl, 9-decenyl, 1-undecenyl, 2-undecenyl, 3-undecenyl, 4-undecenyl, 5-undecenyl, 6-undecenyl, 7-undecenyl, 8-undecenyl, 9-undecenyl, 10-undecenyl, 1-dodecenyl, 2-dodecenyl, 3-dodecenyl, 4-dodecenyl, 5-dodecenyl, 6-dodecenyl, 7-dodecenyl, 8-dodecenyl, 9-dodecenyl, 10-dodecenyl, and 11-dodecenyl. Unless stated otherwise in the specification, an alkyl group can be optionally substituted.

[0044] "Alkynyl" or "alkynyl group" refers to a straight or branched hydrocarbon chain. In embodiments, an alkynyl group contains 1-30 carbon atoms. In embodiments, an alkynyl group contains 2-12 carbon atoms and has one or more carbon-carbon triple bonds, e.g., a straight or branched group of 2-8 carbon atoms, referred to herein as a C2-C8 alkynyl. Each alkynyl group is attached to the remainder of the molecule by a single bond. Alkynyl groups containing any number of carbon atoms from 2 to 12 are included. Alkynyl groups containing up to 12 carbon atoms are C2-C8 alkynyl groups. 12 Alkynyl, containing up to 10 carbon atoms, is C2-C 10Alkynyl, an alkynyl group containing up to 6 carbon atoms is C2-C6 alkynyl, and an alkynyl containing up to 5 carbon atoms is C2-C5 alkynyl. C2-C5 alkynyl includes C5 alkynyl, C4 alkynyl, C3 alkynyl, and C2 alkynyl. C2-C6 alkynyl includes all of the moieties listed above for C2-C5 alkynyl, but also includes C6 alkynyl. C2-C 10 Alkynyl includes all of the moieties listed above for C2-C5 alkynyl and C2-C6 alkynyl, but also includes C7, C8, C9 and C 10 Alkynyl is also included. Similarly, C2-C 12 Alkynyl includes all of the above moieties, except that C 11 and C 12 Alkynyl is also included. C2-C 12 Non-limiting examples of alkenyls include ethynyl, propynyl, butynyl, pentynyl, etc. Unless stated otherwise in the specification, alkyl groups can be optionally substituted.

[0045] "Aryl" refers to a hydrocarbon ring system containing hydrogen, 6-18 carbon atoms, and at least one aromatic ring, attached to the remainder of the molecule by a single bond. For purposes of the present invention, aryl may be a monocyclic, bicyclic, tricyclic, tetracyclic, or other polycyclic ring system, and may include fused or bridged ring systems. Aryl includes, but is not limited to, aryls derived from aceanthrylene, acenaphthylene, acephenanthrylene, anthracene, azulene, benzene, chrysene, fluoranthene, fluorene, as-indacene, s-indacene, indane, indene, naphthalene, phenalene, phenanthrene, preadene, pyrene, and triphenyl. Unless otherwise specified herein, aryls may be optionally substituted.

[0046] An "aralkyl" or "arylalkyl" is a group of the formula -R b -R c R refers to the group bis an alkylene group as defined above, R c is one or more aryl as defined above, e.g., benzyl, diphenylmethyl, etc. Unless stated otherwise in the specification, an aralkyl group can be optionally substituted.

[0047] "Carbocyclyl", "carbocyclic ring" or "carbocycle" refers to a ring structure in which each atom forming the ring is carbon and which is attached to the remainder of the molecule by a single bond. A carbocyclic ring can contain from 3 to 20 carbon atoms in the ring. Carbocyclic rings include aryl, as well as cycloalkyl, cycloalkenyl and cycloalkynyl, as defined herein. Unless otherwise specified herein, a carbocyclyl group can be optionally substituted.

[0048] "Cycloalkyl" refers to a stable non-aromatic monocyclic or polycyclic fully saturated hydrocarbon, consisting only of carbon and hydrogen atoms, which may include fused, spirocyclic, or bridged ring systems, having 3 to 20 carbon atoms and attached to the remainder of the molecule by a single bond. Monocyclic cycloalkyls include, for example, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, and cyclooctyl. Polycyclic cycloalkyls include, for example, adamantyl, norbornyl, decalinyl, 7,7-dimethyl-bicyclo[2.2.1]heptanyl, and the like. Unless otherwise specified in the specification, cycloalkyl groups can be optionally substituted.

[0049] "Cycloalkenyl" refers to a stable non-aromatic monocyclic or polycyclic hydrocarbon consisting solely of carbon and hydrogen atoms and having one or more carbon-carbon double bonds, which may include fused, spirocyclic, or bridged ring systems, having 3-20 carbon atoms, preferably 3-10 carbon atoms, and attached to the remainder of the molecule by a single bond. Monocyclic cycloalkenyls include, for example, cyclopentenyl, cyclohexenyl, cycloheptenyl, cycloctenyl, and the like. Polycyclic cycloalkenyls include, for example, bicyclo[2.2.1]hept-2-enyl, and the like. Unless otherwise stated in the specification, cycloalkenyl groups can be optionally substituted.

[0050] "Cycloalkynyl" refers to a stable non-aromatic monocyclic or polycyclic hydrocarbon consisting solely of carbon and hydrogen atoms, having 3 to 20 carbon atoms, having one or more carbon-carbon triple bonds, and may include fused, spirocyclic, or bridged ring systems, and is attached to the remainder of the molecule by a single bond. Monocyclic cycloalkynyl includes, for example, cycloheptynyl, cyclooctynyl, and the like. Unless otherwise specified in the specification, cycloalkynyl groups can be optionally substituted.

[0051] "Heterocyclyl", "heterocyclic ring" or "heterocycle" refers to a stable 3-20 membered aromatic or non-aromatic ring consisting of 2-12 carbon atoms and 1-6 heteroatoms selected from the group consisting of nitrogen, oxygen, and sulfur. The heterocycle can be aromatic (heteroaryl) or non-aromatic. Unless otherwise stated herein, the heterocyclyl can be a monocyclic, bicyclic, tricyclic, tetracyclic or other polycyclic ring system, which can include fused, spirocyclic, or bridged ring systems; the nitrogen, carbon, or sulfur atoms in the heterocyclyl can be optionally oxidized; the nitrogen atom can be optionally quaternized; the heterocyclyl can be partially or fully saturated. Examples of such heterocyclyls include dioxolanyl, thienyl[1,3]dithianyl, decahydroisoquinolyl, biotinyl, dihydrofuranyl, dihydroindolyl, dihydropyranyl, dihydrothienyl, dithiazolyl, homopiperidinyl, pyranyl, pyrazolinyl, thiopyranyl, imidazolinyl, imidazolidinyl, isothiazolidinyl, isoxazolidinyl, morpholinyl, octahydroindolyl, octahydroisoindolyl, 2-oxopiperazinyl, and the like. Examples of heterocyclyl groups include, but are not limited to, 2-oxopiperidinyl, 2-oxopyrrolidinyl, oxazolidinyl, piperidinyl, piperazinyl, 4-piperidonyl, pyrrolidinyl, pyrrolidin-2-onyl, pyrazolidinyl, quinuclidinyl, thiazolidinyl, tetrahydrofuryl, tetrahydroisoquinolyl, trithianyl, tetrahydropyranyl, thiomorpholinyl, thiamorpholinyl, 1-oxo-thiomorpholinyl, and 1,1-dioxo-thiomorpholinyl. Unless stated otherwise in the specification, heterocyclyl groups can be optionally substituted.

[0052] "Heteroaryl" refers to a 5-20 membered ring system containing a hydrogen atom, 1-13 carbon atoms, 1-6 heteroatoms selected from the group consisting of nitrogen, oxygen and sulfur, and at least one aromatic ring. For purposes of this disclosure, a heteroaryl can be monocyclic, bicyclic, tricyclic or tetracyclic, which can include fused or bridged ring systems; a heteroaryl may contain one or more non-aromatic rings (e.g., cycloalkyl or heterocyclyl) fused to an aromatic ring. The nitrogen, carbon or sulfur atoms in a heteroaryl can be optionally oxidized; the nitrogen atom can be optionally quaternized. Examples include azepinyl, acridinyl, benzimidazolyl, benzothiazolyl, benzoindolyl, benzodioxolyl, benzofuranyl, benzoxazolyl, benzothiazolyl, benzothiadiazolyl, benzo[b][1,4]dioxepinyl, 1,4-benzodioxanyl, benzonaphthofuranyl, benzoxazolyl, benzodioxolyl, benzodioxinyl, benzopyranyl, benzopyranonyl, benzofuranyl, benzofuranonyl, benzothienyl (benzothiophenyl), benzotriazolyl, benzo[4,6]imidazo[1,2-a]pyridinyl, carbazolyl, cinnolinyl, dibenzofuranyl, dibenzothiophenyl, furanyl, furanonyl, isothiazolyl, imidazolyl, indazolyl, indolyl, indazolyl, isoindolyl, indophenyl, and phenyl. These include, but are not limited to, linyl, isoindolinyl, isoquinolyl, indolizinyl, isoxazolyl, naphthyridinyl, oxadiazolyl, 2-oxoazepinyl, oxazolyl, oxiranyl, 1-oxidopyridinyl, 1-oxidopyrimidinyl, 1-oxidopyrazinyl, 1-oxidopyridazinyl, 1-phenyl-1H-pyrrolyl, phenazinyl, phenothiazinyl, phenoxazinyl, phthalazinyl, pteridinyl, purinyl, pyrrolyl, pyrazolyl, pyridinyl, pyrazinyl, pyrimidinyl, pyridazinyl, quinazolinyl, quinoxalinyl, quinolinyl, quinuclidinyl, isoquinolinyl, tetrahydroquinolinyl, thiazolyl, thiadiazolyl, triazolyl, tetrazolyl, triazinyl, and thiophenyl (i.e., thienyl).Unless stated otherwise in the specification, a heteroaryl group can be optionally substituted.

[0053] "Heteroarylalkyl" refers to a group of the formula -R b -R f R refers to the group b is an alkylene chain as defined above, and R f is heteroaryl as defined above. Unless stated otherwise in the specification, a heteroarylalkyl group may be optionally substituted.

[0054] The term "substituted" as used herein means any of the above groups (i.e., alkyl, alkenyl, alkynyl, aryl, arylalkyl, carbocyclyl, cycloalkyl, cycloalkenyl, cycloalkynyl, heterocyclyl, N-heterocyclyl, heteroaryl, etc.) in which at least one hydrogen atom has been replaced by a bond to a non-hydrogen atom, such as, by way of example and not limitation, a halogen atom, such as F, Cl, Br, and I; an oxygen atom in groups such as hydroxyl, alkoxy, and ester groups; a sulfur atom in groups such as thiol, thioalkyl, sulfone, sulfonyl, and sulfoxide groups; a nitrogen atom in groups such as amines, amides, alkylamines, dialkylamines, arylamines, alkylarylamines, diarylamines, N-oxides, imides, and enamines; a silicon atom in groups such as trialkylsilyl, dialkylarylsilyl, alkyldiarylsilyl, and triarylsilyl groups; and other heteroatoms in various other groups. "Substituted" refers to any of the above groups in which one or more hydrogen atoms have been replaced by a higher bond (e.g., a double or triple bond) with a heteroatom, such as oxygen in oxo, carbonyl, carboxyl, and ester groups; and nitrogen in groups such as imine, oxime, hydrazone, and nitrile. For example, "substituted" refers to any of the above groups in which one or more hydrogen atoms have been replaced by a heteroatom, such as oxygen in NR g R h , N.R. g C(=O)R h , N.R.g C(=O)NR g R h , N.R. g C(=O)OR h , N.R. g SO2R h ,OC(=O)NR g R h , OR g , S.R. g , SOR g , SO2R g , OSO2R g , SO2OR g , =NSO2R g , and SO2NR g R h "Substituted" includes any of the above groups replaced with one or more hydrogen atoms. g , C(=O)OR g , C(=O)NR g R h , CH2SO2R g , CH2SO2NR g R h , or any of the above groups substituted with R g and R hare the same or different and are independently hydrogen, alkyl, alkenyl, alkynyl, alkoxy, alkylamino, thioalkyl, aryl, aralkyl, cycloalkyl, cycloalkenyl, cycloalkynyl, cycloalkylalkyl, haloalkyl, haloalkenyl, haloalkynyl, heterocyclyl, N-heterocyclyl, heterocyclylalkyl, heteroaryl, N-heteroaryl, and / or heteroarylalkyl. "Substituted" further refers to any of the above groups, wherein one or more hydrogen atoms are replaced by amino, cyano, hydroxyl, imino, nitro, oxo, thioxo, halo, alkyl, alkenyl, alkynyl, alkoxy, alkylamino, thioalkyl, aryl, aralkyl, cycloalkyl, cycloalkenyl, cycloalkynyl, cycloalkylalkyl, haloalkyl, haloalkenyl, haloalkynyl, heterocyclyl, N-heterocyclyl, heterocyclylalkyl, heteroaryl, N-heteroaryl, and / or heteroarylalkyl groups. In addition, "substituted" refers to any of the above groups in which two hydrogen atoms have been replaced by a bond each to form a fused ring system containing the atom to which the hydrogen is attached. Furthermore, each of the aforementioned substituents can also be optionally substituted with one or more of the above substituents.

[0055] The compounds of the present disclosure contain one or more chiral centers and / or double bonds and therefore may exist as stereoisomers, such as geometric isomers, enantiomers or diastereomers. The term "stereoisomers" as used herein consists of all geometric isomers, enantiomers or diastereomers. These compounds can be designated with the symbols "R" or "S" depending on the arrangement of the substituents around the stereogenic carbon atoms. The present disclosure encompasses various stereoisomers of these compounds and mixtures thereof. Stereoisomers include enantiomers and diastereomers. Mixtures of enantiomers or diastereomers may be designated with "(±)" in the nomenclature, although one of ordinary skill in the art will recognize that the structure may implicitly indicate a chiral center. In embodiments, an enantiomer or stereoisomer may be provided substantially free of the corresponding enantiomer.

[0056] In embodiments, the compound is a racemic mixture of (S)- and (R)-isomers. In other embodiments, provided herein are mixtures of compounds in which the individual compounds of the mixture are present predominantly in the (S)- or (R)-isomeric configuration. For example, the compound mixture has an (S)-enantiomeric excess of greater than about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 95%, about 96%, about 97%, about 98%, about 99%, about 99.5% or more. In other embodiments, the compound mixture has an (S)-enantiomeric excess of about 55% to greater than about 99.5%, about 60% to greater than about 99.5%, about 65% to greater than about 99.5%, about 70% to greater than about 99.5%, about 75% to greater than about 99.5%, about 80% to greater than about 99.5%, about 85% to greater than about 99.5%, about 90% to greater than about 99.5%, about 95% to greater than about 99.5%, about 96% to greater than about 99.5%, about 97% to greater than about 99.5%, about 98% to greater than about 99.5%, about 99% to greater than about 99.5%, or more. In other embodiments, the compound mixture has an (R)-enantiomer purity of greater than about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 95%, about 96%, about 97%, about 98%, about 99%, about 99.5% or more. In some other embodiments, the compound mixture has an (R)-enantiomeric excess of about 55% to greater than about 99.5%, about 60% to greater than about 99.5%, about 65% to greater than about 99.5%, about 70% to greater than about 99.5%, about 75% to greater than about 99.5%, about 80% to greater than about 99.5%, about 85% to greater than about 99.5%, about 90% to greater than about 99.5%, about 95% to greater than about 99.5%, about 96% to greater than about 99.5%, about 97% to greater than about 99.5%, about 98% to greater than about 99.5%, about 99% to greater than about 99.5%, or more.

[0057] Individual stereoisomers of the compounds of the present disclosure can be prepared synthetically from commercially available starting materials containing an asymmetric or stereogenic center, or by preparation of a racemic mixture followed by resolution methods well known to those skilled in the art. These resolution methods are exemplified as follows: (1) coupling of the enantiomeric mixture to a chiral auxiliary, recrystallization or chromatographic separation of the resulting diastereomeric mixture, and liberation of the optically pure product from the auxiliary; (2) salt formation utilizing an optically active resolving agent; or (3) direct separation of the mixture of optical enantiomers on a chiral chromatographic column. Stereoisomeric mixtures can also be resolved into their component stereoisomers by well-known methods such as chiral phase gas chromatography, chiral phase high performance liquid chromatography, crystallization of the compound as a chiral salt complex, or crystallization of the compound in a chiral solvent. Stereoisomers can also be obtained from stereomerically pure intermediates, reagents, and catalysts by well-known asymmetric synthesis methods.

[0058] Geometric isomers may also exist in the compounds of the present disclosure. The present disclosure encompasses various geometric isomers and mixtures thereof resulting from the arrangement of substituents around a carbon-carbon double bond or the arrangement of substituents around a carbon ring. Substituents around a carbon-carbon double bond are designated as being in the "Z" or "E" configuration, and the terms "Z" and "E" are used according to IUPAC standards. Unless otherwise indicated, structures depicting double bonds include both E and Z isomers.

[0059] Substituents around a carbon-carbon double bond may also be referred to as "cis" or "trans", with "cis" referring to substituents on the same side of the double bond and "trans" referring to substituents on opposite sides of the double bond. The arrangement of substituents around a carbon ring is designated as "cis" or "trans". The term "cis" refers to substituents on the same side of the plane of the ring and the term "trans" refers to substituents on opposite sides of the plane of the ring. Mixtures of compounds in which substituents are located both on the same and opposite sides of the plane of the ring are designated as "cis / trans".

[0060] The compounds disclosed herein may exist as tautomers, and both tautomeric forms are intended to be encompassed within the scope of the disclosure, even if only one tautomeric structure is shown.

[0061] As used herein, the term "isotopically enriched variant" is meant to include compounds which differ only in the presence of one or more isotopically enriched atoms. Such compounds may be useful, for example, as analytical tools, probes in biological assays, or as therapeutic agents. For example, an "isotopically enriched variant" of a compound may be an isotopically enriched variant, e.g., of deuterium ( 2 H or D), carbon 13 ( 13 C), nitrogen 15( 15 In such isotopically substituted compounds, the following atoms, if present, may be altered, for example, hydrogen is 2 H / D, carbon 13 C, nitrogen 15 It is understood that the presence and location of such atoms may be N, and that such presence and location can be determined within the skill of the art. Similarly, the present invention may include the preparation of isotopic variants with radioactive isotopes, for example, when the resulting compounds may be used in drug and / or substrate tissue distribution studies. The radioactive isotope tritium, i.e. 3 H, and carbon-14, i.e. 14 C are particularly useful for this purpose due to their ease of incorporation and ready means of detection. Additionally, positron emitting isotopes, e.g. 11 C. 18 F 、15O and 13 Compounds substituted with, for example, N can also be prepared and may be useful in positron emission tomography (PET) experiments to examine substrate receptor occupancy.

[0062] The term "triglyceride" as used herein means an ester derived from glycerol and three fatty acids. The fatty acids may be the same or different. The notation used to describe triglycerides herein is the same as that used to describe fatty acids below. The fatty acids can be attached to the glycerol molecule in any order, e.g., any fatty acid can react with any hydroxyl group of the glycerol molecule to form an ester bond. For example, in a non-limiting example, a triglyceride can include glycerol with any combination of the following fatty acids: C18:1, C14:1, C16:1, polyunsaturated, and saturated. A triglyceride of C18:1 fatty acid simply means that the fatty acid component of the triglyceride is derived from or based on a C18:1 fatty acid. That is, a C18:1 triglyceride is an ester of glycerol with three fatty acids, each having 18 carbon atoms, each fatty acid having one double bond. Similarly, a C14:1 triglyceride is an ester of glycerol and three fatty acids, each having 14 carbon atoms, each fatty acid having one double bond. Similarly, a C16:1 triglyceride is an ester of glycerol and three fatty acids, each having 16 carbon atoms, each fatty acid having one double bond. A combination of a triglyceride of C18:1 fatty acid with a C14:1 and / or C16:1 fatty acid means: (a) a C18:1 triglyceride mixed with a C14:1 triglyceride or a C16:1 triglyceride, or both; or (b) at least one of the fatty acid components of the triglyceride is derived from or based on a C18:1 fatty acid, and the other two are derived from or based on a C14:1 fatty acid and / or a C16:1 fatty acid.

[0063] The term "fatty acid" and similar terms refer to a carboxylic acid having a long aliphatic tail that is saturated or unsaturated. The terms "long aliphatic tail" and "fatty acid chain" are used interchangeably herein. Fatty acids and fatty acid chains can be esterified into phospholipids and triglycerides. As used herein, fatty acid chain lengths include C4-C30 (e.g., C6-C30), saturated or unsaturated, cis or trans, unsubstituted or substituted, branched or unbranched hydrocarbon chains (e.g., fatty acid chain lengths include C4-C30 (e.g., C6-C30), saturated or unsaturated, cis or trans, unsubstituted or substituted with 1-6 side chains). For example, in embodiments, examples of fatty acid chains include, but are not limited to, saturated or unsaturated, cis or trans, unsubstituted or substituted hydrocarbon chains of C4, C5, C6, C7, C8, C9, C10, C11, C12, C13, C14, C15, C16, C17, C18, C19, C20, C21, C22, C23, C24, C25, C26, C27, C28, C29, or C30. Unsaturated fatty acids and fatty acid chains have one or more double bonds between carbon atoms. Saturated fatty acids and fatty acid chains do not contain double bonds. In embodiments, fatty acids are described herein by the capital letter "C" representing a carbon atom, followed by a number representing the number of carbon atoms in the fatty acid, followed by a colon and another number representing the number of double bonds in the fatty acid. For example, C16:1 indicates a fatty acid of 16 carbon atoms with one double bond, e.g., palmitoleic acid. The number after the colon in this notation does not indicate the position of the double bond within the fatty acid, nor does it indicate whether the hydrogen atoms attached to the carbon atoms of the double bond are cis to each other. Other examples of this notation include C18:0 (stearic acid), C18:1 (oleic acid), C18:2 (linoleic acid), C18:3 (α-linolenic acid), and C20:4 (arachidonic acid).

[0064] The term "sterol", for example, but not limited to, cholesterol, can also be utilized in the methods and compounds described herein. Sterols are animal or plant steroids that contain only a hydroxyl group at C-3 and no other functional groups. Generally, sterols contain 27-30 carbon atoms and one double bond at the 5 / 6 position, and occasionally at the 7 / 8, 8 / 9 or other positions. Besides these unsaturated species, other sterols are saturated compounds obtained by hydrogenation. One example of a suitable animal sterol is cholesterol. Typical examples of suitable phytosterols that are preferred from an application point of view are ergosterol, campesterol, stigmasterol, brassicasterol, and preferably sitosterol or sitostanol, more specifically β-sitosterol or β-sitostanol. Besides the phytosterols mentioned, their esters are preferably used. The acid component of the ester may be reduced to a carboxylic acid corresponding to the formula (CA-I): RI CO-OH (CA-I); where RI CO is an aliphatic linear or branched acyl group containing 2-30 carbon atoms and O and / or 1, 2 or 3 double bonds. Typical examples are acetic acid, propionic acid, hexanoic acid, butyric acid, valeric acid, caproic acid, caprylic acid, 2-ethylhexanoic acid, capric acid, cyclopentanepropionic acid, lauric acid, isotridecanoic acid, myristic acid, palmitic acid, palmitoleic acid, stearic acid, isostearic acid, oleic acid, elaidic acid, petroselinic acid, linoleic acid, conjugated linoleic acid (CLA), linolenic acid, elaeosteric add, arachic acid, gadoleic acid, behenic acid and erucic acid.

[0065] The term "phospholipid" refers to an amphipathic compound consisting of two hydrophobic fatty acid "tails," and a hydrophilic "head" consisting of a phosphate group. The two components are linked by a glycerol molecule. The phosphate group can be modified with simple organic molecules such as choline, ethanolamine, or serine. Choline refers to an essential bioactive nutrient with the chemical formula R-(CH2)2-N(CH2)4. When the phosphorylated portion is R-, it is called phosphocholine.

[0066] As used herein, "lysolipid" includes, in non-limiting embodiments (acyl, single chain), by way of example, 1-myristoyl-2-hydroxy-sn-glycero-3-phosphocholine (MHPC), 1-palmitoyl-2-hydroxy-sn-glycero-3-phosphocholine (PHPC), and 1-stearoyl-2-hydroxy-sn-glycero-3-phosphocholine (SHPC).

[0067] The terms "apolipoprotein AI" or "apo AI" and also "apoliprotein A1" or "apoA1" refer to the protein that in humans is encoded by the APOAI gene.

[0068] Detailed Description Classically, the vertebrate immune system is subdivided into two parts. The first part, the innate immunity, provides an initial response to infection within minutes to hours. Its cellular components include natural killer (NK) cells, innate lymphoid cells (ILCs), and phagocytes such as monocytes, macrophages, and neutrophils. The innate immune system is triggered through the recognition of either pathogen or endogenous danger signals by pattern recognition receptors (PRRs) and acts as a rapid first line of defense. Upon detection of pathogen-associated molecular patterns (PAMPs), PRRs initiate the innate immune response, which includes antigen presentation, costimulation, and subsequent activation of the adaptive immune system through cytokine excretion. In addition, PRRs also recognize damage-associated molecular patterns (DAMPs), which trigger non-infectious inflammatory responses. The second phase of the response to infection involves the adaptive response, the second part of the immune system, in which T and B lymphocytes specifically recognize, proliferate, and are activated against a pathogen. These cells also build an immunological memory for that specific infection. The specificity of the adaptive immune system response is mediated by recombination of immunoglobulin genes at the lymphocyte level. Immunological memory results in a more rapid and quantitatively superior immune response (compared to only a primary response) to previously encountered antigens. Although it was long thought that the innate immune system had no memory, recent studies have shown that innate immune cells undergo metabolic and epigenetic rewiring to adjust their functional programs in a process called "trained immunity," which can be considered a de facto innate immune memory.

[0069] Trained immunity is defined by a secondary, long-term hyperresponsiveness, manifested by increased cytokine secretion triggered by metabolic and epigenetic rewiring to restimulation of myeloid cells and their progenitors and stem cells in bone marrow, spleen, and blood after a primary insult. Trained immunity (also called innate immune memory) is also defined by a long-term increased responsiveness (e.g., high cytokine production) after restimulation of myeloid innate immune cells with a secondary stimulus, induced by a primary insult priming these cells or their progenitors and stem cells in bone marrow and spleen, mediated by epigenetic, metabolic, and transcriptional rewiring.

[0070] Trained immunity is regulated and maintained through the induction of training properties in progenitor cells in the bone marrow, resulting in persistent reprogramming that extends beyond the lifespan of bone marrow cells in the bloodstream. Trained immunity can be induced in cultured bone marrow cells using a variety of "training agents," but its systemic induction requires the participation of bone marrow progenitor cells.

[0071] In one aspect, the present disclosure provides compounds that activate nucleotide-binding oligomerization domain-containing protein 2 (NOD2) (e.g., compounds of formula (I), (IA), (IB), (II), (II-1), (II-2), (IIA), (IIA-1), (IIA-2) or Table 1). The present disclosure also provides nanobiological compositions comprising nanoparticle carriers (e.g., HDL-derived nanoparticles) comprising a compound of the present disclosure (e.g., of formula (I), (IA), (IB), (II), (II-1), (II-2), (IIA), (IIA-1), (IIA-2) or Table 1). Nanobiological compositions of the present disclosure comprising compounds of the present disclosure (e.g., of formula (I), (IA), (IB), (II), (II-1), (II-2), (IIA), (IIA-1), (IIA-2) or Table 1) that activate nucleotide-binding oligomerization domain-containing protein 2 (NOD2) are designed to exhibit bone marrow proclivity. These nanomaterials can be administered (e.g., intravenously) to promote trained immunity. Therapeutic induction of trained immunity may find use, for example, in overcoming immune paralysis in sepsis and infection, treating cell proliferation disorders (such as cancer), enhancing immune responses, and the like.

[0072] compound In embodiments, the present disclosure provides a compound of formula (I):

[0073] [ka] or a pharma- ceutically acceptable salt thereof. (In the formula, R 1 is -H or -C(O)-R X and; R 2 and R 3 are each independently selected from the group consisting of -H, alkyl, alkylene-aryl, -C(O)-alkyl, and -C(O)-aryl; R 4 , R 5 , and R 5’ are each alkyl; R 6 and R 11 are each independently -H or alkyl; R 7 is the fatty acid chain, -YN(R 6 )-C(O)-O-alkylene-C(H)(OR 8 )-Alkylene-OR 9 , -YN(R 6 )-C(O)-R X , -YOP(O)(OH)-O-alkylene-C(H)(OR 8 )-Alkylene-OR 9 or -Y-triazolyl-L; Y is alkylene; L is a fatty acid chain, -alkylene-C(O)-W, -alkylene-OC(O)-W, -alkylene-N-(alkylene-C(O)-NR 11 -Alkylene-NR 11 -C(O)-W)2, and -alkylene-N-(alkylene-C(O)-W)2; W is a fatty acid chain, -O-alkylene-C(H)(OR 8 )-Alkylene-OR 9 , a phospholipid, or a sterol; R 8 and R 9 are each independently R X or -C(O)-R X and; R 22 , R 33 , R 33’ , R 44 , R 44’ , R 55 , and R 55’ are each independently H or R A and; R X is a fatty acid chain; Each of the foregoing alkyl, alkylene, alkylene-aryl, aryl, and triazolyl may be selected from one or more R A Optionally replaced by R A is hydrogen, halo, alkoxy, haloalkoxy, cyano, hydroxyl, -N(RC )(R D ), -C(O)N(R C )(R D ), -N(R C )C(O)R B , -OC(O)NR C R D , -NR C C(O)OR B , -OC(O)R B , -C(O)OR B , -C(O)R B , -CO2H, -NO2, -SH, S(O) X R B (wherein X is 0, 1, or 2), aryl, heteroaryl, heteroarylalkyl, cycloalkyl, heterocycloalkyl, and R B independently selected for each occurrence from the group consisting of: R C and R D is hydrogen, alkyl, haloalkyl-C(O)R B , and -C(O)OR B or R C and R D together with the nitrogen to which they are attached, R A forming an optionally substituted heterocycle; R B is alkyl, alkenyl, or alkynyl optionally substituted with one or more fluoro. to provide.

[0074] In embodiments, the present disclosure provides a compound of formula (I):

[0075] [ka] or a pharma- ceutically acceptable salt thereof. (In the formula, R 1 is -H or -C(O)-R X and; R 2 and R 3are each independently selected from the group consisting of -H, alkyl, alkylene-aryl, -C(O)-alkyl, and -C(O)-aryl; R 4 , R 5 , and R 5’ are each alkyl; R 6 and R 11 are each independently -H or alkyl; R 7 is C 9~30 Fatty acid chain, -YN(R 11 )-C(O)-O-alkylene-C(H)(OR 8 )-Alkylene-OR 9 , -C(R 10 )(C(O)NH2)-alkylene-N(R 11 )-C(O)-C 16~30 Fatty acid chain, -(CR 10 R 10 )2-OP(O)(OH)-O-alkylene-C(R 10 )(OR Z )-Alkylene-OR Z’ or -Y-triazolyl-L; R Z and R Z’ are each independently C 8~30 Fatty acid chain or -C(O)-C 16~30 is a fatty acid chain; Y is alkylene; L is a fatty acid chain, -alkylene-C(O)-W, -alkylene-OC(O)-W, -alkylene-N-(alkylene-C(O)-NR 11 -Alkylene-NR 11 -C(O)-W)2, and -alkylene-N-(alkylene-C(O)-W)2; W is a fatty acid chain, -O-alkylene-C(H)(OR 8 )-Alkylene-OR 9 , a phospholipid, or a sterol; R 8 and R 9 are each independently R X or -C(O)-R Xand; R 10 , R 22 , R 33 , R 33’ , R 44 , R 44’ , R 55 , and R 55’ are each independently H or R A and; R X is a fatty acid chain; Each of the foregoing alkyl, alkylene, alkylene-aryl, aryl, and triazolyl may be selected from one or more R A Optionally replaced by R A is hydrogen, halo, alkoxy, haloalkoxy, cyano, hydroxyl, -N(R C )(R D ), -C(O)N(R C )(R D ), -N(R C )C(O)R B , -OC(O)NR C R D , -NR C C(O)OR B , -OC(O)R B , -C(O)OR B , -C(O)R B , -CO2H, -NO2, -SH, S(O) X R B (wherein X is 0, 1, or 2), aryl, arylalkyl, heteroaryl, heteroarylalkyl, cycloalkyl, heterocycloalkyl, and R B independently selected for each occurrence from the group consisting of: R C and R D is hydrogen, alkyl, haloalkyl-C(O)R B , and -C(O)OR B or R C and R D together with the nitrogen to which they are attached, R A forming an optionally substituted heterocycle; R Bis alkyl, alkenyl, or alkynyl optionally substituted with one or more fluoro; R 7 C 9~30 If it is a fatty acid chain, R 2 is -H) to provide.

[0076] In an embodiment of the compound of formula (I), the compound has the formula (IA):

[0077] [ka] (In the formula, R 1 is -H or -C(O)-R X and; R 2 and R 3 are each independently selected from the group consisting of -H, alkyl, alkylene-aryl, -C(O)-alkyl, and -C(O)-aryl; R 4 , R 5 , and R 5’ are each alkyl; R 6 and R 11 are each independently -H or alkyl; R 7 is C 9~30 Fatty acid chain, -YN(R 11 )-C(O)-O-alkylene-C(H)(OR 8 )-Alkylene-OR 9 , -C(R 10 )(C(O)NH2)-alkylene-N(R 11 )-C(O)-C 16~30 Fatty acid chain, -(CR 10 R 10 )2-OP(O)(OH)-O-alkylene-C(R 10 )(OR Z )-Alkylene-OR Z’ or -Y-triazolyl-L; R Z and R Z’ are each independently C 8~30Fatty acid chain or -C(O)-C 16~30 is a fatty acid chain; Y is alkylene; L is a fatty acid chain, -alkylene-C(O)-W, -alkylene-OC(O)-W, -alkylene-N-(alkylene-C(O)-NR 11 -Alkylene-NR 11 -C(O)-W)2, and -alkylene-N-(alkylene-C(O)-W)2; W is a fatty acid chain, -O-alkylene-C(H)(OR 8 )-Alkylene-OR 9 , a phospholipid, or a sterol; R 8 and R 9 are each independently R X or -C(O)-R X and; R 10 , R 22 , R 33 , R 33’ , R 44 , R 44’ , R 55 , and R 55’ are each independently H or R A and; R X is a fatty acid chain; Each of the foregoing alkyl, alkylene, alkylene-aryl, aryl, and triazolyl may be selected from one or more R A Optionally replaced by R A is hydrogen, halo, alkoxy, haloalkoxy, cyano, hydroxyl, -N(R C )(R D ), -C(O)N(R C )(R D ), -N(R C )C(O)R B , -OC(O)NR C R D , -NR C C(O)OR B , -OC(O)R B , -C(O)OR B , -C(O)R B, -CO2H, -NO2, -SH, S(O) X R B (wherein X is 0, 1, or 2), aryl, arylalkyl, heteroaryl, heteroarylalkyl, cycloalkyl, heterocycloalkyl, and R B independently selected for each occurrence from the group consisting of: R C and R D is hydrogen, alkyl, haloalkyl-C(O)R B , and -C(O)OR B or R C and R D together with the nitrogen to which they are attached, R A forming an optionally substituted heterocycle; R B is alkyl, alkenyl, or alkynyl optionally substituted with one or more fluoro; R 7 C 9~30 If it is a fatty acid chain, R 2 is -H) or a pharma- ceutically acceptable salt thereof.

[0078] In an embodiment, the compound of formula (I) has the formula (IB):

[0079] [ka] or a pharma- ceutically acceptable salt thereof.

[0080] In an embodiment of the compound of formula (I), (IA), or (IB), Y is -C(O)N(R C )(R D In embodiments, Y is an alkylene optionally substituted with -C 1~6 In an embodiment, Y is -CH-. In an embodiment, Y is -CH- or

[0081] [ka] In an embodiment, Y is

[0082] [ka] In an embodiment, Y is

[0083] [ka] It is.

[0084] In an embodiment of the compound of formula (I), (IA), or (IB), R 7 is -Y-triazolyl-L.

[0085] In an embodiment of the compound of formula (I), (IA), or (IB), R 7 comprises a cholesteryl moiety or at least one fatty acid chain containing at least 17 carbons. 7 comprises a cholesteryl moiety. In embodiments, R 7 comprises at least one fatty acid chain containing at least 17 carbons. In an embodiment, R 7 comprises at least two fatty acid chains containing at least 17 carbons. 7 At least two C 17 In embodiments, the C 17 The fatty acid chain is derived from stearic acid or oleic acid. 17 The fatty acid chain is derived from stearic acid. 17 The fatty acid chain is derived from oleic fatty acid. 7 is a 2-C ester derived from stearic acid. 17 Contains fatty acid chains.

[0086] In an embodiment of the compound of formula (I), (IA), or (IB), R 7 is an alkyl group having at least 16 carbons. In embodiments of the compounds of formula (I), (IA), or (IB), R7 is an alkenyl group having at least 16 (carbons). In embodiments of the compounds of formula (I), (IA), or (IB), R 7 is an alkyl group having at least 18 carbons. In an embodiment of the compound of formula (I), R 7 is an alkenyl group having at least 18 carbons.

[0087] In an embodiment of the compound of formula (I), (IA), or (IB), R 7 is C 9~30 In an embodiment of the compound of formula (I), R 7 -C 9~30 Alkyl or C 9~30 alkenyl. In embodiments, R 7 -C 9~30 Alkyl or C 9~30 alkenyl, where R 7 -C 9~30 For alkyl, R 2 is -H. In an embodiment of the compound of formula (I), R 7 -C 9~30 In embodiments, R 7 -C 9~30 is alkyl, R 2 is -H. In an embodiment of the compound of formula (I), R 7 -C 9~30 alkenyl. In embodiments, R 7 -C 15~30 In embodiments, R 7 -C 15~30 is an alkyl group, R 2 is -H. In embodiments, R 7 -C 15~30 In embodiments, R 7 -C 17~19 In embodiments, R 7 -C 17~19 is alkyl, R 2 is -H. In embodiments, R 7 -C 17~19 alkenyl. In embodiments, R7 -C 18 In embodiments, R 7 -C 18 is an alkyl group, R 2 is -H. In embodiments, R 7 -C 18 It is an alkenyl group.

[0088] In an embodiment of the compound of formula (I), (IA), or (IB), R 7 teeth,

[0089] [ka] It is.

[0090] In an embodiment of the compound of formula (I), (IA), or (IB), R 7 teeth,

[0091] [ka] It is.

[0092] In an embodiment of the compound of formula (I), (IA), or (IB), R 7 teeth,

[0093] [ka] and R 2 is -H.

[0094] In an embodiment of the compound of formula (I), (IA), or (IB), R 7 is -YN(R 11 )-C(O)-O-alkylene-C(H)(OR 8 )-Alkylene-OR 9 It is.

[0095] In an embodiment of the compound of formula (I), (IA), or (IB), R 7 is -C(R 10)(C(O)NH2)-alkylene-N(R 11 )-C(O)-C 16~30 In an embodiment, R 7 is -C(H)(C(O)NH2)-C5 alkylene-N(R 11 )-C(O)-C 17~30 In an embodiment, R 7 is -C(H)(C(O)NH2)-alkylene-N(R 11 )-C(O)-C 17~30 It is a fatty acid.

[0096] In an embodiment of the compound of formula (I), (IA), or (IB), R 7 is -C(R 10 )(C(O)NH2)-alkylene-N(R 11 )-C(O)-C 16~30 In embodiments, R 7 is -C(R 10 )(C(O)NH2)-alkylene-N(R 11 )-C(O)-C 17~30 In embodiments, R 7 is -C(H)(C(O)NH2)-alkylene-N(R 11 )-C(O)-C 17~30 In embodiments, R 7 is -C(H)(C(O)NH2)-C4 alkylene-N(R 11 )-C(O)-C 17~30 In an embodiment of the compound of formula (I), R 7 is -C(R 10 )(C(O)NH2)-alkylene-N(R 11 )-C(O)-C 16~30 is alkyl, R 2 is alkylene-aryl (e.g., benzyl). In embodiments, R 7 is -C(R 10 )(C(O)NH2)-alkylene-N(R 11 )-C(O)-C 17~30 is alkyl, R 2 is alkylene-aryl (e.g., benzyl). In embodiments, R 7is -C(H)(C(O)NH2)-alkylene-N(R 11 )-C(O)-C 17~30 is alkyl, R 2 is alkylene-aryl (e.g., benzyl). In embodiments, R 7 is -C(H)(C(O)NH2)-C4 alkylene-N(R 11 )-C(O)-C 17~30 is alkyl, R 2 is alkylene-aryl (eg, benzyl).

[0097] In an embodiment of the compound of formula (I), (IA), or (IB), R 7 is -(CR 10 R 10 )2-OP(O)(OH)-O-alkylene-C(R 10 )(OR Z )-Alkylene-OR Z In an embodiment of the compound of formula (I), R 7 is -CH2CH2-OP(O)(OH)-O-CH2-C(H)(OR Z )-CH2-OR Z’ In an embodiment, R Z and R Z’ are each independently C 12~20 Alkyl or -C(O)-C 16~30 In an embodiment, R Z and R Z’ are each independently C 18 Alkyl or -C(O)-C 17 In embodiments, R Z and R Z’ are each independently -C 16~30 In embodiments, R Z and R Z’ are both -C 17 It is an alkyl.

[0098] In an embodiment of the compound of formula (I), (IA), or (IB), R 7 is -YN(R 6 )-C(O)-O-alkylene-C(H)(OR8 )-Alkylene-OR 9 In an embodiment, R 8 and R 9 are each independently 8~30 Alkyl or -C(O)-C 8~30 In embodiments, R 8 and R 9 are each independently 12~20 Alkyl or -C(O)-C 11~20 In embodiments, R 8 and R 9 are each independently 18 Alkyl or -C(O)-C 17 In embodiments, R 8 and R 9 are both -C(O)-C 17 It is an alkyl.

[0099] In an embodiment of the compound of formula (I), (IA), or (IB), R A is hydrogen, halo, alkoxy, haloalkoxy, cyano, hydroxyl, -N(R C )(R D ), -C(O)N(R C )(R D ), -N(R C )C(O)R B , -OC(O)NR C R D , -NR C C(O)OR B , -OC(O)R B , -C(O)OR B , -C(O)R B , -CO2H, -NO2, -SH, S(O) X R B wherein X is 0, 1, or 2, and is independently selected at each occurrence from the group consisting of aryl, arylalkyl, heteroaryl, heteroarylalkyl, cycloalkyl, and heterocycloalkyl.

[0100] In an embodiment of the compound of formula (I), (IA), or (IB), R Ais halo, alkoxy, haloalkoxy, cyano, hydroxyl, -N(R C )(R D ), -C(O)N(R C )(R D ), -N(R C )C(O)R B , -OC(O)NR C R D , -NR C C(O)OR B , -OC(O)R B , -C(O)OR B , -C(O)R B , -CO2H, -NO2, -SH, S(O) X R B wherein X is 0, 1, or 2, and is independently selected at each occurrence from the group consisting of aryl, arylalkyl, heteroaryl, heteroarylalkyl, cycloalkyl, and heterocycloalkyl.

[0101] In an embodiment, the compound of formula (I) has formula (II):

[0102] [ka] or a pharma- ceutically acceptable salt thereof. (In the formula, X1 is -N- and X2 is -C-; or X1 is -C- and X2 is -N-; R 2 and R 3 are each independently selected from the group consisting of -H, alkyl, aryl, alkylene-aryl, -C(O)-alkyl, and -C(O)-aryl; R 4 , R 5 , and R 5’ are each alkyl; R 6 and R 11 are each independently -H or alkyl; Y is alkylene; L is a fatty acid chain, -alkylene-C(O)-W, -alkylene-OC(O)-W, -alkylene-N-(alkylene-C(O)-NR 11 -Alkylene-NR 11 -C(O)-W)2, and -alkylene-N-(alkylene-C(O)-W)2; W is a fatty acid chain, -O-alkylene-C(H)(OR 8 )-Alkylene-OR 9 , a phospholipid, or a sterol; R 8 and R 9 are each independently R X or -C(O)-R X and; R X is a fatty acid chain; Each of the foregoing alkyl, alkylene, alkylene-aryl, and aryl may be selected from one or more R A and is optionally replaced by R A is hydrogen, halo, alkoxy, haloalkoxy, cyano, hydroxyl, -N(R C )(R D ), -C(O)N(R C )(R D ), -N(R C )C(O)R B , -OC(O)NR C R D , -NR C C(O)OR B , -OC(O)R B , -C(O)OR B , -C(O)R B , -CO2H, -NO2, -SH, S(O) X R B (wherein X is 0, 1, or 2), aryl, arylalkyl, heteroaryl, heteroarylalkyl, cycloalkyl, heterocycloalkyl, and R B independently selected for each occurrence from the group consisting of: R C and R D is hydrogen, alkyl, haloalkyl-C(O)R B , and -C(O)ORB or R C and R D together with the nitrogen to which they are attached, R A forming an optionally substituted heterocycle; R B is alkyl, alkenyl, or alkynyl optionally substituted with one or more fluoro. It is.

[0103] In an embodiment, the compound of formula (II) is

[0104] [ka] It is.

[0105] In an embodiment, the compound of formula (II) has the formula (II-1):

[0106] [ka] or a pharma- ceutically acceptable salt thereof.

[0107] In an embodiment, the compound of formula (II-1) is

[0108] [ka] or a pharma- ceutically acceptable salt thereof.

[0109] In an embodiment, the compound of formula (II) has the formula (II-2):

[0110] [ka] It is a compound of the formula:

[0111] In an embodiment, the compound of formula (II-2) is

[0112] [ka] or a pharma- ceutically acceptable salt thereof.

[0113] In an embodiment, the compound of formula (I) has the formula (IIA):

[0114] [ka] or a pharma- ceutically acceptable salt thereof. (In the formula, X1 is -N- and X2 is -C-; or X1 is -C- and X2 is -N-; R 2 and R 3 are each independently selected from the group consisting of -H, alkyl, aryl, alkylene-aryl, -C(O)-alkyl, and -C(O)-aryl; R 4 , R 5 , and R 5’ are each alkyl; R 6 and R 11 are each independently -H or alkyl; Y is alkylene; L is a fatty acid chain, -alkylene-C(O)-W, -alkylene-OC(O)-W, -alkylene-N-(alkylene-C(O)-NR 11 -Alkylene-NR 11 -C(O)-W)2, and -alkylene-N-(alkylene-C(O)-W)2; W is a fatty acid chain, -O-alkylene-C(H)(OR 8 )-Alkylene-OR 9 , a phospholipid, or a sterol; R 8 and R 9 are each independently R X or -C(O)-R X and; R X is a fatty acid chain; Each of the foregoing alkyl, alkylene, alkylene-aryl, and aryl may be selected from one or more R A and is optionally replaced by R A is hydrogen, halo, alkoxy, haloalkoxy, cyano, hydroxyl, -N(R C )(R D ), -C(O)N(R C )(R D ), -N(R C )C(O)R B , -OC(O)NR C R D , -NR C C(O)OR B , -OC(O)R B , -C(O)OR B , -C(O)R B , -CO2H, -NO2, -SH, S(O) X R B (wherein X is 0, 1, or 2), aryl, arylalkyl, heteroaryl, heteroarylalkyl, cycloalkyl, heterocycloalkyl, and R B independently selected for each occurrence from the group consisting of: R C and R D is hydrogen, alkyl, haloalkyl-C(O)R B , and -C(O)OR B or R C and R D together with the nitrogen to which they are attached, R A forming an optionally substituted heterocycle; R B is alkyl, alkenyl, or alkynyl optionally substituted with one or more fluoro. It is.

[0115] In an embodiment, the compound of formula (IIA) is

[0116] [ka] It is.

[0117] In an embodiment, the compound of formula (IIA) is a compound of formula (IIA-1), or a pharma- ceutically acceptable salt thereof.

[0118] [ka]

[0119] In an embodiment, the compound of formula (IIA-1) is

[0120] [ka] or a pharma- ceutically acceptable salt thereof.

[0121] In an embodiment, the compound of formula (IIA) is a compound of formula (IIA-2), or a pharma- ceutically acceptable salt thereof.

[0122] [ka]

[0123] In an embodiment, the compound of formula (IIA-2) is

[0124] [ka] or a pharma- ceutically acceptable salt thereof.

[0125] In an embodiment of the compound of (I), (IA), (IB), (II), (II-1), (II-2), (II-A), (IIA-1) or (IIA-2), Y is alkylene. In an embodiment of the compound of (I), (IA), (IB), (II), (II-1), (II-2), (II-A), (IIA-1) or (IIA-2), Y is C 1~6 In an embodiment, Y is C 1~5 In an embodiment, Y is C 1~3In embodiments, Y is -C(O)N(R C )(R D ), and R C and R D is defined herein. In an embodiment, Y is -CH2-. In an embodiment, Y is

[0126] [ka] In an embodiment, Y is

[0127] [ka] It is.

[0128] In an embodiment of compound (I), (IA), (IB), (II), (II-1), (II-2), (II-A), (IIA-1) or (IIA-2), R A is hydrogen, halo, alkoxy, haloalkoxy, cyano, hydroxyl, -N(R C )(R D ), -C(O)N(R C )(R D ), -N(R C )C(O)R B , -OC(O)NR C R D , -NR C C(O)OR B , -OC(O)R B , -C(O)OR B , -C(O)R B , -CO2H, -NO2, -SH, S(O) X R B (wherein X is 0, 1, or 2), aryl, arylalkyl, heteroaryl, heteroarylalkyl, cycloalkyl, heterocycloalkyl, and R B and independently selected at each occurrence from the group consisting of:

[0129] In an embodiment of compound (I), (IA), (IB), (II), (II-1), (II-2), (II-A), (IIA-1) or (IIA-2), R A is halo, alkoxy, haloalkoxy, cyano, hydroxyl, -N(R C )(R D ), -C(O)N(R C )(R D ), -N(R C )C(O)R B , -OC(O)NR C R D , -NR C C(O)OR B , -OC(O)R B , -C(O)OR B , -C(O)R B , -CO2H, -NO2, -SH, S(O) X R B (wherein X is 0, 1, or 2), aryl, arylalkyl, heteroaryl, heteroarylalkyl, cycloalkyl, heterocycloalkyl, and R B and independently selected at each occurrence from the group consisting of:

[0130] In embodiments of the compounds of (I), (IA), (IB), (II), (II-1), (II-2), (II-A), (IIA-1) or (IIA-2), the alkyl, alkylene, alkylene-aryl and aryl are each independently selected from one or more R A is optionally replaced by

[0131] In an embodiment of compound (I), (IA), (IB), (II), (II-1), (II-2), (II-A), (IIA-1) or (IIA-2), R C and R D is hydrogen, alkyl, haloalkyl-C(O)R B , and -C(O)OR B or R C and R D together with the nitrogen to which they are attached, R A forming an optionally substituted heterocycle.

[0132] In an embodiment of compound (I), (IA), (IB), (II), (II-1), (II-2), (II-A), (IIA-1) or (IIA-2), R B is alkyl, alkenyl, or alkynyl optionally substituted with one or more fluoro.

[0133] In an embodiment of compound (I), (IA), (IB), (II), (II-1), (II-2), (II-A), (IIA-1) or (IIA-2), R A is -H.

[0134] In some embodiments of compounds (I), (IA), (IB), (II), (II-1), (II-2), (II-A), (IIA-1) or (IIA-2), R 2 is alkyl, aryl, or alkylene-aryl. In embodiments, the aryl is optionally substituted with alkyl.

[0135] In an embodiment of compound (I), (IA), (IB), (II), (II-1), (II-2), (II-A), (IIA-1) or (IIA-2), R 2 and R 3 are each independently selected from the group consisting of -H, alkyl, aryl, alkylene-aryl, -C(O)-alkyl, and -C(O)-aryl.

[0136] In an embodiment of compound (I), (IA), (IB), (II), (II-1), (II-2), (II-A), (IIA-1) or (IIA-2), R 2 is -H or benzyl. In an embodiment of the compound of formula (I), R 2 -H ’ In an embodiment, R 2 is benzyl.

[0137] In an embodiment of compound (I), (IA), (IB), (II), (II-1), (II-2), (II-A), (IIA-1) or (IIA-2), R 4 , R 5 , and R 5’ are each alkyl.

[0138] In an embodiment of compound (I), (IA), (IB), (II), (II-1), (II-2), (II-A), (IIA-1) or (IIA-2), R 4 is alkyl. In embodiments, R 4 is methyl.

[0139] In an embodiment of compound (I), (IA), (IB), (II), (II-1), (II-2), (II-A), (IIA-1) or (IIA-2), R 3 is -H. In embodiments, R 6 is -H. In embodiments, R 3 and R 6 are both -H.

[0140] In an embodiment of compound (I), (IA), (IB), (II), (II-1), (II-2), (II-A), (IIA-1) or (IIA-2), R 10 , R 22 , R 33 , R 33’ , R 44 , R 44’ , R 55 , and R 55’ are -H, respectively.

[0141] In an embodiment of the compound of formula (I), (II), (IA), (IB), (II-1), (II-2), (II-A), (IIA-1) or (IIA-2), L comprises a cholesteryl moiety or at least one fatty acid chain comprising at least 13 carbons. In an embodiment, L comprises a cholesteryl moiety. In an embodiment, L comprises at least one fatty acid chain comprising at least 13 carbons. In an embodiment, L comprises at least two fatty acid chains comprising at least 15 carbons. In an embodiment, L comprises at least one C 17 In an embodiment, L comprises at least two C 17 In an embodiment, L is C 17 Alkyl or C 17 In an embodiment, L comprises at least one fatty acid chain independently selected from the group consisting of C, C- ... 17 Alkyl or C 17 In embodiments, the fatty acid chains are independently selected from C, C- ... 17 The fatty acid chain is derived from stearic acid or oleic acid. 17 The fatty acid chain is derived from stearic acid. 17 The fatty acid chain is derived from oleic fatty acid. In an embodiment, L is a 2 C fatty acid chain derived from stearic acid. 17 Contains fatty acid chains.

[0142] In an embodiment of the compound of (I), (IA), (IB), (II), (II-1), (II-2), (II-A), (IIA-1) or (IIA-2), L is a fatty acid chain, -alkylene-C(O)-W, -alkylene-OC(O)-W, -alkylene-N-(alkylene-C(O)-NR 11 -Alkylene-NR 11 -C(O)-W)2, and -alkylene-N-(alkylene-C(O)-W)2.

[0143] In an embodiment of the compound of (I), (IA), (IB), (II), (II-1), (II-2), (II-A), (IIA-1) or (IIA-2), L is C8~30 Fatty acid chain, -CH2-C(O)-W, -CH2-OC(O)-W, -CH2CH2-N-CH2CH2-C(O)-NR 11 -CH2CH2-NR 11 -C(O)-W)2, and -CH2CH2-N-(CH2CH2-C(O)-W)2.

[0144] In an embodiment of the compound of (I), (IA), (IB), (II), (II-1), (II-2), (II-A), (IIA-1) or (IIA-2), L is C 8~30 In an embodiment, L is a C 8~30 Alkyl or C 8~30 In an embodiment, L is C 15~20 Alkyl or C 15~20 It is alkenyl.

[0145] In an embodiment of the compound of (I), (IA), (IB), (II), (II-1), (II-2), (II-A), (IIA-1) or (IIA-2), L is -alkylene-C(O)-W, -alkylene-OC(O)-W, -alkylene-N-(alkylene-C(O)-NR 11 -Alkylene-NR 11 -C(O)-W)2, or -alkylene-N-(alkylene-C(O)-W)2.

[0146] In an embodiment of the compound of (I), (IA), (IB), (II), (II-1), (II-2), (II-A), (IIA-1) or (IIA-2), L is -alkylene-C(O)-W. In an embodiment, L is -C 1~6 Alkylene-C(O)-W. In embodiments, L is -CH2-C(O)-W.

[0147] In an embodiment of the compound of (I), (IA), (IB), (II), (II-1), (II-2), (II-A), (IIA-1) or (IIA-2), L is -alkylene-OC(O)-W. In an embodiment, L is -C 1~6Alkylene -OC(O)-W. In embodiments, L is -CH2-OC(O)-W.

[0148] In an embodiment of the compound of (I), (IA), (IB), (II), (II-1), (II-2), (II-A), (IIA-1) or (IIA-2), L is -alkylene-N-(alkylene-C(O)-NR 11 -Alkylene-NR 11 In an embodiment, L is -C(O)-W). 2~6 Alkylene-N-(-C 2-6 Alkylene-C(O)-NR 11 -C 2~6 Alkylene-NR 11 In an embodiment, L is -CH-CH-N-(CH-CH-C(O)-NR 11 -CH2-CH2-NR 11 -C(O)-W)2.

[0149] In an embodiment of the compound of (I), (IA), (IB), (II), (II-1), (II-2), (II-A), (IIA-1) or (IIA-2), L is -alkylene-N-(alkylene-C(O)-W). In an embodiment, L is -C 1~6 Alkylene-N-(C 1~6 alkylene-C(O)-W). In embodiments, L is -CH-CH-N-(CH-CH-C(O)-W).

[0150] In an embodiment of the compound of (I), (IA), (IB), (II), (II-1), (II-2), (II-A), (IIA-1) or (IIA-2), L is C 18 In an embodiment, L is a C 18 Alkyl or C 18 alkenyl. In embodiments, L is -CH2(CH2CH2)8-CH3.

[0151] In an embodiment of the compound of (I), (IA), (IB), (II), (II-1), (II-2), (II-A), (IIA-1) or (IIA-2), W is a fatty acid chain, -O-alkylene-C(H)(OR 8 )-Alkylene-OR 9 , phospholipids, or sterols.

[0152] In an embodiment of the compound of (I), (IA), (IB), (II), (II-1), (II-2), (II-A), (IIA-1) or (IIA-2), W is C 8~30 In an embodiment, W is a C 8~30 Alkyl or C 8~30 In an embodiment, W is C 8~30 In an embodiment, W is C 8~30 In an embodiment, W is C 12~18 In an embodiment, W is a C 12~18 Alkyl or C 12~18 In an embodiment, W is C 12~18 In an embodiment, W is C 12~18 In an embodiment, W is C 18 In an embodiment, W is a C 17 In an embodiment, W is a C 17 Alkyl or C 17 In an embodiment, W is C 17 In an embodiment, W is C 17 In an embodiment, W is an alkenyl. In an embodiment, W is -(CH2CH2)8-CH3. In an embodiment, W is a fatty acid chain containing at least 15 carbons. In an embodiment, W is a fatty acid chain containing at least 18 carbons. In an embodiment, W is a fatty acid chain containing at least 17 carbons. In an embodiment, W is a fatty acid chain containing at least 18 carbons.

[0153] In an embodiment of the compound of (I), (IA), (IB), (II), (II-1), (II-2), (II-A), (IIA-1) or (IIA-2), W is

[0154] [ka] In an embodiment, R X and R X’ are each independently a fatty acid chain. X and R X’ are each independently a fatty acid chain containing at least 15 carbons. X and R X’ are each independently a fatty acid chain containing at least 17 carbons. X and R X’ are each independently -C 8~30 In an embodiment, R X and R X’ are each independently -C 8~30 Alkyl or -C 8~30 alkenyl. In embodiments, R X and R X’ are both -C 8~30 In embodiments, R X and R X’ are both -C 8~30 alkenyl. In embodiments, R X and R X’ are each independently C 12~18 In an embodiment, R X and R X’ are each independently -C 12~18 Alkyl or -C 12~18 alkenyl. In embodiments, R X and R X’ -C 12~18 In embodiments, R X and R X’ -C 12~18 alkenyl. In embodiments, R X and R X’ are each independently C 17 In an embodiment, R X and R X’ are each independently C17 Alkyl or C 17 alkenyl. In embodiments, R X and R X’ is C 17 In embodiments, R X and R X’ is C 17 In embodiments, C is alkenyl. 17 The chains are each independently derived from stearic acid or oleic acid. 17 The chain is derived from stearic acid. 17 The chain is derived from oleic acid. X and R X’ are both -(CH2CH2)8-CH3.

[0155] In an embodiment of compound (I), (IA), (IB), (II), (II-1), (II-2), (II-A), (IIA-1) or (IIA-2), W is a sterol.

[0156] In an embodiment of compound (I), (IA), (IB), (II), (II-1), (II-2), (II-A), (IIA-1) or (IIA-2), W is cholesterol.

[0157] [ka]

[0158] In an embodiment of the compound (I), (IA), (IB), (II), (II-1), (II-2), (II-A), (IIA-1) or (IIA-2), W is a phospholipid selected from the group consisting of phosphatidylcholine (PC), phosphatidylglycerol (PG), phosphatidylserine (PS), phosphatidylethanolamine (PE), phosphatidic acid (PA), and lysophosphatidylcholine. In an embodiment, W is phosphatidylcholine (PC). In an embodiment, W is phosphatidylglycerol (PG). In an embodiment, W is phosphatidylserine (PS). In an embodiment, W is phosphatidylethanolamine (PE). In an embodiment, W is phosphatidic acid (PA). In an embodiment, W is lysophosphatidylcholine.

[0159] In an embodiment of the compound of (I), (IA), (IB), (II), (II-1), (II-2), (II-A), (IIA-1) or (IIA-2), W is

[0160] [ka] It is. (In the formula, Y Q1 , Y Q2 , and Y Q3 are each independently alkylene. Q1 is C 2~6 is alkylene, and Y Q2 and Y Q3 are each independently -C 1~3 In embodiments, R X and R X’ are each independently a fatty acid chain having at least 15 carbons, or in certain embodiments, R X and R X’ are each independently a fatty acid chain having at least 17 carbons. X and R X’ are each independently C 8~30 In an embodiment, RX and R X’ are each independently C 8~30 Alkyl or C 8~30 alkenyl. In embodiments, R X and R X’ are each independently C 15~30 Alkyl or C 15~30 alkenyl. In embodiments, R X and R X’ are each independently C 15~20 Alkyl or C 15~20 alkenyl. In embodiments, R X and R X’ are each independently C 17 Alkyl or C 17 alkenyl. In embodiments, R X and R X’ are both -(CH2CH2)8-CH3.

[0161] In an embodiment of the compound of (I), (IA), (IB), (II), (II-1), (II-2), (II-A), (IIA-1) or (IIA-2), W is

[0162] [ka] It is.

[0163] In an embodiment of the compound of (I), (IA), (IB), (II), (II-1), (II-2), (II-A), (IIA-1) or (IIA-2), W is

[0164] [ka] or a pharma- ceutically acceptable salt thereof. (In the formula, R X and R X’ are each independently C 8~30 In an embodiment, the fatty acid is saturated. X and R X’ are each independently C8~30 Alkyl or C 8~30 alkenyl. In embodiments, R X and R X’ are each independently C 15~20 Alkyl or C 15~20 alkenyl. In embodiments, R X and R X’ are each independently C 17 Alkyl or C 17 alkenyl. In embodiments, R X and R X’ are both -(CH2CH2)8-CH3.

[0165] In an embodiment of compound (I), (IA), (IB), (II), (II-1), (II-2), (II-A), (IIA-1) or (IIA-2), R 8 and R 9 are each independently R X or -C(O)-R X It is.

[0166] In an embodiment of compound (I), (IA), (IB), (II), (II-1), (II-2), (II-A), (IIA-1) or (IIA-2), R 6 and R 11 are each independently -H or alkyl.

[0167] In an embodiment, the present disclosure provides a method for producing a pharmaceutical composition comprising:

[0168] [ka]

[0169] [ka] A compound selected from the group consisting of: or a stereoisomer thereof (eg, an alpha or beta anomer thereof, or a tautomer thereof).

[0170] In an embodiment, the present disclosure provides a method for producing a method for treating a cancer cell comprising:

[0171] [ka] A compound selected from the group consisting of: or a stereoisomer thereof (eg, an anomer thereof, or a mixture of anomers thereof).

[0172] In embodiments, the disclosure provides a compound of formula (I), (IA), (IB), (II), (II-1), (II-2), (IIA), (IIA-1), (IIA-2), or a stereoisomer thereof.

[0173] In embodiments, the disclosure provides a compound of formula (I), (IA), (IB), (II), (II-1), (II-2), (IIA), (IIA-1), (IIA-2), or a diastereomer or tautomer thereof.

[0174] In embodiments, provided herein is one or more compounds selected from Table 1.

[0175] In embodiments, provided herein is one or more pharma- ceutically acceptable salts of a compound selected from Table 1.

[0176] In embodiments, provided herein is one or more compounds selected from Table 1, stereoisomers, or pharma- ceutically acceptable salts thereof.

[0177] [Table 1-1]

[0178] [Table 1-2]

[0179] [Table 1-3]

[0180] [Table 1-4]

[0181] [Table 1-5]

[0182] Muramyl tripeptide phosphatidylethanolamine; N-(N-acetylmuramoyl)-L-alanyl-D-alpha-glutaminyl-N-[(7R)-4-hydroxy-4-oxido-10-oxo-7-[(1-oxohexadecyl)oxy]-3,5,9-trioxa-4-phosphapentacos-1-yl]-L-alaninamide (MTP-a-DPPE or mifamurtide): Molecular weight: 1238 Daltons. CLogP=10.59 (uncharged) and 4.80 (negatively charged). Mifamurtide (CAS number [83461-56-7]) was prepared according to literature procedures (e.g., DE; Wade, R. (1985) J Label Compd Radiopharm. 22 (1): 29-35. doi:10.1002 / jlcr.2580220105). The lipophilicity of this molecule is relatively low with a CLogP of 4.80 in physiological environments.

[0183] N-Acetylmuramyl-L-alanyl-D-isoglutamine-6-O-stearoyl (MDP-C18[mur]) molecular weight: 759 Daltons. CLogP=5.39 (uncharged) and 1.39 (negatively charged). MDP-C18[mur] (CAS number [60398-08-5]) was prepared according to literature procedures (e.g., Matsumoto K. et al. (1981) Infect Immun. 32(2):748-58). The lipophilicity of this molecule is low in physiological environments, with a CLogP of 1.39, and is therefore unlikely to be sufficient to ensure reliable incorporation into HDL-derived nanoparticles.

[0184] Romurtide (CAS number [78113-36-7]) Molecular weight: 887 Daltons. It has lipophilicity with CLogP=3.90 (uncharged) and 0.61 (negatively charged) (CLogP 0.61 in physiological environment). This compound has low lipophilicity, with a CLogP close to 0 for charged molecules.

[0185] Murabutide (CAS number [74817-61-1]) Molecular weight: 549 Daltons. CLogP=-1.53 ​​(uncharged) and CLogP value is negative. This molecule is hydrophilic since its CLogP value is less than 0.

[0186] In embodiments, a compound of the disclosure (e.g., of Formula (I), (IA), (IB), (II), (II-1), (II-2), (IIA), (IIA-1), (IIA-2) or Table 1) activates nucleotide-binding oligomerization domain-containing protein 2 (NOD2).

[0187] Anomers and open / closed structures In embodiments, the molecules of the present disclosure bear an -OH substituent at the anomeric hemiacetal carbon of the muramyl sugar group, i.e., when R2=H, it is understood that both anomeric isomers alpha and beta are included in the compounds of the present disclosure.

[0188] Furthermore, in those cases where R2 = H, it is known in the art that such molecules actually exist (in an aqueous environment) in both closed and open ring isomers, and again, it is understood that both open and closed ring isomers are included in the compounds of the present disclosure.

[0189] In the non-limiting examples below, the top structures show the alpha and beta anomers, and the bottom structures show the general anomeric closed (left) and open (right) ring structures.

[0190] [ka]

[0191] molecular weight The compounds of the invention preferably have a molecular weight of greater than 500 daltons, greater than 700 daltons, greater than 950 daltons, or greater than 1,200 daltons.

[0192] The compounds of the invention preferably have a molecular weight of less than 10,000 daltons, less than 5,000 daltons, less than 2,500 daltons, or less than 1,750 daltons.

[0193] Hydrophobicity In certain embodiments, the compounds of the present disclosure are hydrophobic in nature.Hydrophobicity can be estimated by calculating CLogP value.This can be carried out in software programs such as Perkin Elmer's ChemDraw or ChemDraw Professional (v18).The higher the CLogP value of a compound, the more hydrophobic the compound is.

[0194] In embodiments, compounds of the present disclosure have a CLogP value of greater than about 1, greater than about 3, greater than about 5, greater than about 7, greater than about 9, or greater than about 11.

[0195] The CLogP value represents the n-octanol / water partition coefficient (Log Po / w) of a molecule, which is a calculated value as opposed to a LogP value, i.e. an experimentally evaluated value. Therefore, CLogP values ​​may deviate from LogP values. Importantly, however, CLogP values ​​provide a good comparison between the lipophilicity of molecules. CLogP values ​​can be evaluated for molecules in either the uncharged or charged state. This is the case for molecules with ionogenic groups, such as those with carboxylic acid (-COOH) or phosphate (-OP(O)OH-O-) groups. At physiological pH (approximately 7.4), these particular groups are deprotonated and therefore charged. Also, alkyl(ated)amine groups are charged at physiological pH, in this case by protonation.

[0196] At physiological pH, the molecules of the invention have CLogP values ​​of less than 20, or less than 15, or less than 10. Additionally, at physiological pH, the molecules of the invention have CLogP values ​​of greater than 3, or greater than 4, or greater than 5, or greater than 5.5.

[0197] Nanobiological Compositions Provided herein are nanobiological compositions comprising a nanoparticle carrier and one or more compounds of the present disclosure (such as compounds (I), (IA), (IB), (II), (II-1), (II-2), (II-A), (IIA-1) or (IIA-2) as disclosed herein or in Table 1).

[0198] In embodiments, the compounds of the present disclosure can be formulated in nanoparticle carriers, including colloidal dispersion systems such as polyplexes, polymer complexes, nanocapsules, microspheres, beads, and lipid-based systems such as oil-in-water emulsions, micelles, mixed micelles, liposomes, lipoplexes, lipid nanoparticles, lipid nanocapsules, lipidoids, rapid clearing lipid nanoparticles (reLNPs), micro- and nanoemulsions, HDL-derived nanoparticles, polymeric nanoparticles such as poly(lactic-co-glycolic acid) (PLGA) nanoparticles, including PLGA microspheres, poly(lactide) (PLA) nanoparticles, poly(ε-caprolactone) (PCL) nanoparticles, poly(butyl cyanoacrylate) (PBCA) nanoparticles, dendrimers (de mdrimers), hyperbranched polyglycerol (HPG) nanoparticles, PEG-polyaspartic acid micelle nanoparticles, cationic polymers such as poly(L-lysine), polyethyleneimine (PEI), DEAE-dextran, poly(amino ester) (PBAE) and chitosan, cyclodextrin nanoparticles, metal nanoparticles, surfactant-based emulsions, virus-like particles (e.g., particles that are primarily composed of viral structural proteins but are non-infectious or have low infectivity), peptide, or protein-based particles, such as albumin nanoparticles, nanowires, gold nanoparticles, magnetic nanoparticles, core-shell nanoparticles, carbon nanotubes, nanocrystals, hyaluronidase, and combinations thereof.

[0199] In embodiments, the compounds of the present disclosure may be formulated in nanoparticle carriers such as those described in U.S. Pat. Nos. 5,567,434, 5,552,157, 5,565,213, 5,738,868, 5,795,587, 10,485,884, U.S. Patent Publication No. 2018 / 0263907, U.S. Patent Publication No. 2016 / 0317647, U.S. Patent Publication No. 2019 / 0290593, U.S. Patent Publication No. 2020 / 0253884, U.S. Patent Publication No. 2020 / 0376146, and WO 2018 / 071549, the contents of each of which are incorporated herein by reference.

[0200] In an embodiment, the nanoparticle carrier is a high density lipoprotein (HDL)-derived nanoparticle. High density lipoprotein (HDL)-derived nanoparticles are envisioned as delivery vehicles that improve the therapeutic index of, for example, small molecule immunomodulatory compounds and / or provide innate immune cell-specific delivery. By providing target specificity to innate immune cells (such as bone marrow cells, bone marrow progenitor cells, hematopoietic stem cells in bone marrow, blood, and / or spleen), therapeutic agents encapsulated or incorporated in HDL-derived nanoparticles can be delivered in a focused and localized manner. In an embodiment, the high density lipoprotein (HDL)-derived nanoparticles comprise apoA-I or a peptidomimetic of apoA-I. In an embodiment, the high density lipoprotein (HDL)-derived nanoparticles comprise apoA-I.

[0201] Human apoA-I can be isolated or prepared by any method known in the art. In an embodiment, human apoA-I is isolated from human HDL. Another known method includes synthesis of apoA-I by recombinant protein expression, for example in Escherichia coli (E. coli) organisms. When expressed in bacteria, apoA-I may contain an N-terminal methionine or formylmethionine. The presence of methionine groups can be assessed by mass spectrometry (MS) methods known in the art. The location of methionine in the protein sequence can be assessed by digestion of apoA-I followed by analysis of the peptide mixture by MS, as also known in the art.

[0202] In embodiments, purification of apoA-I, including any of its variations, may involve any method known in the art (e.g., use of hydrophobic interaction chromatography, ion exchange columns, precipitation, etc.) The production method may or may not include the use of affinity tags that allow for purification of the protein, such tags need to be removed after purification to restore the identity of human apoA-I.

[0203] In embodiments, the high density lipoprotein (HDL)-derived nanoparticles comprise ApoA-1 Milano.

[0204] Suitable apoA-I mimetic polypeptides can have the sequences shown in Table 2 (SEQ ID NOs:256-263, and 342-346) or SEQ ID NOs:1-341.

[0205] [Table 2]

[0206] In embodiments, the apoA-I mimetic is DWLKAFYDKVAEKLKEAF (SEQ ID NO: 256). In embodiments, the apoA-I mimetic is Ac-DWLKAFYDKVAEKLKEAF-NH2 (SEQ ID NO: 257). In embodiments, the apoA-I mimetic is Ac-DWFKAFYDKVAEKFKEAF-NH2 (SEQ ID NO: 260).

[0207] In embodiments, the apoA-I mimic is optionally acetylated at the N-terminus or optionally amidated at the C-terminus. In embodiments, the apoA-I mimic is acetylated at the N-terminus. In embodiments, the apoA-I mimic is amidated at the C-terminus. In embodiments, the apoA-I mimic is acetylated at the N-terminus and amidated at the C-terminus. In embodiments, the HDL-derived nanoparticles of the present disclosure comprise one or more phospholipids. All phospholipids ranging in chain length from C4 to C30, saturated or unsaturated, cis or trans, unsubstituted or substituted with 1-6 side chains, and with or without the addition of lysolipids are contemplated for use in the nanoparticles described herein. Additionally, other synthetic variants and variants with other phospholipid head groups are also contemplated. In embodiments, the HDL-derived nanoparticles comprise phospholipids. In embodiments, the HDL-derived nanoparticles comprise phospholipids and lysolipids.

[0208] Non-limiting examples of phospholipids that can be used in the present composition include phosphatidylcholine (PC), phosphatidylglycerol (PG), phosphatidylserine (PS), phosphatidylethanolamine (PE).In embodiments, phosphatidic acid / ester (PA) can be used.

[0209] In embodiments, the phospholipid or lysolipid is one or more of the following: DDPC CAS-3436-44-0 1,2-didecanoyl-sn-glycero-3-phosphocholine, DEPA-NA CAS-80724-31-8 1,2-dierucoyl-sn-glycero-3-phosphate (sodium salt), DEPC CAS-56649-39-9 1,2-dierucoyl-sn-glycero-3-phosphocholine, DEPE CAS-988-07-2 1,2-dierucoyl-sn-glycero-3-phosphoethanolamine, DEPG-NA 1,2-dierucoyl-sn-glycero-3-phospho-rac-(1-glycerol) (sodium salt), DLOPC CAS-998-06-1 1,2-dilinoleoyl-sn-glycero-3-phosphocholine, DLPA-NA 1,2-Dilauroyl-sn-glycero-3-phosphate (sodium salt), DLPC CAS-18194-25-7 1,2-Dilauroyl-sn-glycero-3-phosphocholine, DLPE 1,2-Dilauroyl-sn-glycero-3-phosphoethanolamine, DLPG-NA 1,2-Dilauroyl-sn-glycero-3-phospho-rac-(1-glycerol) (sodium salt), DLPG-NH4 1,2-Dilauroyl-sn-glycero-3-phospho-rac-(1-glycerol) (ammonium salt), DLPS-NA 1,2-Dilauroyl-sn-glycero-3-phosphoserine (sodium salt), DMPA-NA CAS-80724-3 1,2-Dimyristoyl-sn-glycero-3-phosphate (sodium salt), DMPC CAS-18194-24-6 1,2-Dimyristoyl-sn-glycero-3-phosphocholine, DMPE CAS-988-07-2 1,2-Dimyristoyl-sn-glycero-3-phosphoethanolamine, DMPG-NA CAS-67232-80-8 1,2-Dimyristoyl-sn-glycero-3-phospho-rac-(1-glycerol)(sodium salt), DMPG-NH4 1,2-Dimyristoyl-sn-glycero-3-phospho-rac-(l-glycerol)(ammonium salt), DMPG-NH4 / NA 1,2-Dimyristoyl-sn-glycero-3-phospho-rac-(l-glycerol)(sodium / ammonium salt), DMPS-NA1,2-Dimyristoyl-sn-glycero-3-phosphoserine (sodium salt), DOPA-NA 1,2-Dioleoyl-sn-glycero-3-phosphate (sodium salt), DOPC CAS-4235-95-4 1,2-Dioleoyl-sn-glycero-3-phosphocholine, DOPE CAS-4004-5-1 1,2-Dioleoyl-sn-glycero-3-phosphoethanolamine, DOPG-NA CAS-62700-69-0 1,2-Dioleoyl-sn-glycero-3-phospho-rac-(1-glycerol) (sodium salt), DOPS-NA CAS-70614-14-1 1,2-Dioleoyl-sn-glycero-3-phosphoserine (sodium salt), DPPA-NA CAS-71065-87-7 1,2-Dipalmitoyl-sn-glycero-3-phosphate (sodium salt), DPPC CAS-63-89-8 1,2-Dipalmitoyl-sn-glycero-3-phosphocholine, DPPE CAS-923-61-5 1,2-Dipalmitoyl-sn-glycero-3-phosphoethanolamine, DPPG-NA CAS-67232-81-9 1,2-Dipalmitoyl-sn-glycero-3-phospho-rac-(l-glycerol) (sodium salt), DPPG-NH4 CAS-73548-70-6 1,2-Dipalmitoyl-sn-glycero-3-phospho-rac-(l-glycerol) (ammonium salt), DPPS-NA 1,2-Dipalmitoyl-sn-glycero-3-phosphoserine (sodium salt), DSPA-NA CAS-108321-18-2 1,2-distearoyl-sn-glycero-3-phosphate (sodium salt), DSPC CAS-816-94-4 1,2-distearoyl-sn-glycero-3-phosphocholine, DSPE CAS-1069-79-0 1,2-distearoyl-sn-glycero-3-phosphoethanolamine, DSPG-NA CAS-67232-82-0 1,2-distearoyl-sn-glycero-3-phospho-rac-(1-glycerol) (sodium salt), DSPG-NH4 CAS-108347-80-4 1,2-distearoyl-sn-glycero-3-phospho-rac-(1-glycerol) (ammonium salt), DSPS-NA1,2-distearoyl-sn-glycero-3-phosphoserine (sodium salt), EPC egg-PC, HEPC hydrogenated egg-PC, HSPC hydrogenated soy PC, LYSOPC MYRISTIC CAS-18194-24-6 l-myristoyl-sn-glycero-3-phosphocholine, LYSOPC PALMITIC CAS-17364-16-8 l-palmitoyl-sn-glycero-3-phosphocholine, LYSOPC STEARIC CAS-19420-57-6 l-stearoyl-sn-glycero-3-phosphocholine, milk sphingomyelin, MPPC l-myristoyl-2-palmitoyl-sn-glycero-3-phosphocholine, MSPC l-myristoyl-2-stearoyl-sn-glycero-3-phosphocholine, PMPC 1-Palmitoyl-2-myristoyl-sn-glycero-3-phosphocholine, POPC CAS-26853-31-6 l-Palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine, POPE 1-Palmitoyl-2-oleoyl-sn-glycero-3-phosphoethanolamine, POPG-NA CAS-81490-05-3 l-Palmitoyl-2-oleoyl-sn-glycero-3[phospho-rac-(1-glycerol)](sodium salt), PSPC 1-Palmitoyl-2-stearoyl-sn-glycero-3-phosphocholine, SMPC 1-Stearoyl-2-myristoyl-sn-glycero-3-phosphocholine, SOPC1-Stearoyl-2-oleoyl-sn-glycero-3-phosphocholine, SPPC1-Stearoyl 1-2-palmitoyl-sn-glycero-3-phosphocholine. In some preferred embodiments, non-limiting examples of phospholipids include dimyristoyl phosphatidylcholine (DMPC), soy lecithin, dipalmitoyl phosphatidylcholine (DPPC), distearoyl phosphatidylcholine (DSPC), dilauryl phosphatidylcholine (DLPC), dioleoyl phosphatidylcholine (DOPC), dilaurylolyl phosphatidylglycerol (DLPG), dimyristoyl phosphatidylglycerol (DMPG), dipalmitoyl phosphatidylglycerol (DPPG), distearoyl phosphatidylglycerol (DSPG ...laurylolyl phosphatidylglycerol (DMPG), dipalmitoyl phosphatidylglycerol (DPPG), distearoyl phosphatidylglycerol (DSPG), dilaurylolyl phosphatidylglycerol (DSPG), dilaurylolyl phosphatidylglycerol (DSPG), dilaurylolyl phosphatidylglycerol (DSPG), dilaurylolyl phosphatidylglycerol (DSPG), dilaurylolyl phosphatidylglycerol (DSPG), dilaurylolyl phosphatidylglycerol (DSPG), dilaurylol Oleoylphosphatidylglycerol (DOPG), dimyristoyl phosphatidic acid (DMPA), dimyristoyl phosphatidic acid (DMPA), dipalmitoyl phosphatidic acid (DPPA), dipalmitoyl phosphatidic acid (DPPA), dimyristoyl phosphatidylethanolamine (DMPE), dipalmitoyl phosphatidylethanolamine (DPPE), dimyristoyl phosphatidylserine (DMPS), dipalmitoyl phosphatidylserine (DPPS), dipalmitoyl sphingomyelin (DPSP), distearoyl sphingomyelin (DSSP), and mixtures thereof.

[0210] In embodiments, the phospholipid is 1,2-dimyristoyl-sn-glycero-3-phosphatidylcholine (DMPC) and the lysolipid is 1-myristoyl-2-hydroxy-sn-glycero-phosphocholine (MHPC).

[0211] In embodiments, the phospholipid is 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC) and the lysolipid is 1-palmitoyl-2-hydroxy-sn-glycero-3-phosphocholine (PHPC).

[0212] In embodiments, when the composition comprises (or consists essentially of, or consists of) two or more types of lipids (such as phospholipids, or lysolipids), the weight ratio of the two types of phospholipids ranges from about 1:10 to about 10:1, e.g., about 1:9, about 1:8, about 1:7, about 1:6, about 1:5, about 1:4, about 1:3, about 1:2, about 1:1, about 2:1, about 3:1, about 4:1, about 5:1, about 6:1, about 7:1, about 8:1, about 9:1 to about 10:1, including all values ​​and ranges therebetween.

[0213] In embodiments, the HDL-derived nanoparticles comprise DMPC and MHPC, and the weight ratio of DMPC to MHPC may range from about 1:10 to about 10:1, about 2:1 to about 4:1, about 1:1 to about 5:1, about 2:1 to about 5:1, about 6:1 to about 10:1, about 7:1 to about 10:1, about 8:1 to about 10:1, about 7:1 to about 9:1, or about 8:1 to about 9:1. The weight ratio of DMPC to MHPC may be about 1:10, about 1:9, about 1:8, about 1:7, about 1:6, about 1:5, about 1:4, about 1:3, about 1:2, about 1:1, about 2:1, about 3:1, about 4:1, about 5:1, about 6:1, about 7:1, about 8:1, about 9:1, or about 10:1, including all values ​​and ranges therebetween.

[0214] In embodiments, the HDL-derived nanoparticles comprise POPC and PHPC, and the weight ratio of POPC to PHPC may range from about 1:10 to about 10:1, about 2:1 to about 4:1, about 1:1 to about 5:1, about 2:1 to about 5:1, about 6:1 to about 10:1, about 7:1 to about 10:1, about 8:1 to about 10:1, about 7:1 to about 9:1, or about 8:1 to about 9:1. The weight ratio of POPC to PHPC may be about 1:10, about 1:9, about 1:8, about 1:7, about 1:6, about 1:5, about 1:4, about 1:3, about 1:2, about 1:1, about 2:1, about 3:1, about 4:1, about 5:1, about 6:1, about 7:1, about 8:1, about 9:1, or about 10:1.

[0215] In embodiments, the phospholipids in the nanoparticles of the present disclosure comprise (or consist essentially of, or consist of) a mixture of double-chain diacyl phospholipids and single-chain acyl phospholipids / lysolipids.

[0216] In embodiments, the high density lipoprotein (HDL)-derived nanoparticles comprise apoA-I or a peptidomimetic of apoA-I, and a phospholipid. In embodiments, the high density lipoprotein (HDL)-derived nanoparticles comprise apoA-I or a peptidomimetic of apoA-I, a phospholipid, and a compound of formula (I), (II), (II-1), (II-2), (II-A), (IIA-1) or (IIA-2).

[0217] In embodiments, the high density lipoprotein (HDL) derived nanoparticles comprise i) apoA-I or a peptidomimetic of apoA-I; ii) a phospholipid; iii) a lysolipid, and iv) cholesterol. In embodiments, the high density lipoprotein (HDL) derived nanoparticles comprise i) apoA-I or a peptidomimetic of apoA-I; ii) a phospholipid; iii) a lysolipid, iv) cholesterol, and a compound of formula (I), (II), (II-1), (II-2), (II-A), (IIA-1) or (IIA-2).

[0218] In embodiments, the high density lipoprotein (HDL)-derived nanoparticles comprise i) apoA-I or a peptidomimetic of apoA-I; ii) phospholipids; and iii) cholesterol. In embodiments, the high density lipoprotein (HDL)-derived nanoparticles comprise i) apoA-I or a peptidomimetic of apoA-I; ii) phospholipids; iii) cholesterol; and a compound of formula (I), (II), (II-1), (II-2), (II-A), (IIA-1) or (IIA-2).

[0219] In embodiments, the high density lipoprotein (HDL) derived nanoparticles comprise i) apoA-I or a peptidomimetic of apoA-I; ii) phospholipids; iii) lysolipids, iv) a hydrophobic matrix core, and v) cholesterol. In embodiments, the high density lipoprotein (HDL) derived nanoparticles comprise i) apoA-I or a peptidomimetic of apoA-I; ii) phospholipids; iii) lysolipids, iv) a hydrophobic matrix core, v) cholesterol and a compound of formula (I), (II), (II-1), (II-2), (II-A), (IIA-1) or (IIA-2).

[0220] In embodiments, the high density lipoprotein (HDL)-derived nanoparticles comprise i) apoA-I or a peptidomimetic of apoA-I; ii) a phospholipid; iii) a lysolipid; iv) a triglyceride; v) cholesterol and a compound of formula (I), (II), (II-1), (II-2), (II-A), (IIA-1) or (IIA-2).

[0221] In embodiments, the structure and properties (e.g., particle size, stiffness, viscosity, loading amount, etc.) of HDL-derived nanoparticles can be modified by incorporating hydrophobic matrix.As used herein, hydrophobic matrix refers to the core, filler, or structure modifier of nanobiological preparations.Non-limiting examples of suitable hydrophobic matrix molecules include triglycerides, fatty acid esters, hydrophobic polymers, sterol esters, or combinations thereof.

[0222] For example, the inclusion of one or more triglycerides and / or one or more polymers in the nanoparticles disclosed herein can facilitate the control of nanoparticle size (e.g., from about 10 nm to more than 100 nm) and shape (disk-shaped to spherical). The size, stiffness, and viscosity of the HDL-derived nanoparticles can also affect the loading and biodistribution. In a non-limiting example, HDL-derived nanoparticles containing phospholipids and apoA-I can have a diameter of about 10 nm to about 50 nm, and the addition of hydrophobic matrix molecules (such as triglycerides) causes the HDL-derived nanoparticles to swell from a minimum of about 10 nm to at least about 30 nm. The addition of further triglycerides can further increase the diameter of the HDL-derived nanoparticles to at least 50 nm, at least 75 nm, at least 100 nm, at least 150 nm, at least 200 nm, at least 300 nm, and up to 400 nm, including all values ​​and ranges therebetween.

[0223] Any suitable synthetic or natural fatty acid or fatty acid ester known in the art is contemplated for use in the HDL-derived nanoparticles of the present disclosure. Non-limiting examples of fatty acids that may be used include arachidonic acid, oleic acid, arachidic acid, lauric acid, sad, capric acid, myristic acid, palmic acid, stearic acid, linoleic acid, linolenic acid, dicapric acid, decanoin 3, glycerol mono fatty acid ester, dilaurin, 1-Sunsoft 767, laurocapram (1-dodecyl-aza-cycloheptane-2-ketone), acylcarnitine, acylcholine or C1-C2 fatty acid ester. 10 arrcostab (such as isopropyl myristate IPM), monoglycerides, diglycerides or pharma- ceutically acceptable salts thereof.

[0224] Any suitable synthetic or natural triglyceride known in the art is contemplated for use in the HDL-derived nanoparticles of the present disclosure.Non-limiting examples of triglycerides that can be used include: tricaprylin, tristearin, triolein, tripalmitin, 1,2-dipalmitolein, 1,3-dipalmitolein, 1-palmito-3-stearo-2-olein, 1-palmito-2-stearo-3-olein, 2-palmito-1-stearo-3-olein, trilinolein, 1,2-dipalmitrinolein, 1-palmito-dilinolein, 1-stearo-dilinolein, 1,2-diacetopalmitin, 1,2-distearo-olein, 1,3-distearo-olein, trimyristin, trilaurin and combinations thereof.Suitable triglycerides can be added to the composition of the present invention in raw form. Additionally or alternatively, suitable triglyceride-containing oils and / or processed oils may be added to the composition.Non-limiting examples of oils include coconut oil, corn germ oil, olive oil, palm seed oil, cottonseed oil, palm oil, rapeseed oil, sunflower oil, whale oil, soybean oil, peanut oil, linseed oil, tall oil, and combinations thereof.

[0225] The hydrophobic polymer or polymers can be selected from the group of polymers approved for human use (i.e., biocompatible and FDA approved). Such polymers include, for example, but are not limited to, the following polymers, derivatives of such polymers, copolymers, block copolymers, branched polymers, and polymer blends: polyalkene dicarboxylates, polyanhydrides, poly(aspartic acid), polyamides, polybutylene succinate (PBS), polybutylene succinate-co-adipate (PBSA), poly(ε-caprolactone) (PCL), polycarbonates including polyalkylene carbonates (PC), aliphatic polyesters, and polyesteramides. These include polyesters, polyethylene succinate (PES), polyglycolide (PGA), polyimines and polyalkyleneimines (Pl, PAI), polylactides (PLA (polylactic acid), PLLA, PDLLA), polylactic-co-glycolic acid (PLGA), poly(L-lysine), polymethacrylates, polypeptides, polyorthoesters, poly-p-dioxanone (PPDO), (hydrophobically) modified polysaccharides, polysiloxanes and polyalkylsiloxanes, polyureas, polyurethanes, polyvinyl alcohol, and biodegradable polyalkylcyanoacrylates.

[0226] In embodiments of the HDL-derived nanoparticles of the present disclosure, the addition of cholesterol to the nanoparticle carrier stabilizes the composition and improves entrapment efficiency. Typically, HDL-derived nanoparticles contain about 1 mol% to about 100 mol% cholesterol relative to the phospholipid (e.g., relative to DMPC), including all ranges and values ​​therebetween, such as about 1 mol%, about 2 mol%, about 3 mol%, about 4 mol%, about 5 mol%, about 6 mol%, about 7 mol%, about 8 mol%, about 9 mol%, about 10 mol%, about 11 mol%, about 12 mol%, about 13 mol%, about 14 mol%, about 15 mol%, about 16 mol%, about 17 mol%, about 18 mol%, about 19 mol%, about 20 mol%, about 21 mol%, about 22 mol%, about 23 mol%, about 24 mol%, about 25 mol%, about 26 mol%, about 27 mol%, about 28 mol%, about 29 mol%, about 30 mol%, about 31 mol%, about 32 mol%, about 33 mol%, about 34 mol%, about 35 mol%, about 36 mol%, about 37 mol%, about 38 mol%, about 39 mol%, about 40 mol%, about 41 mol%, about 42 mol%, about 43 mol%, about 44 mol%, about 45 mol%, about 46 mol%, about 47 mol%, about 48 mol%, about 49 mol%, about 50 mol%, about 51 mol%, about 52 mol%, about 53 mol%, about 54 mol%, about 55 mol%, about 56 mol%, about 57 mol%, about 58 mol%, about 59 mol%, about 60 mol%, about %, about 22 mol%, about 23 mol%, about 24 mol%, about 25 mol%, about 26 mol%, about 27 mol%, about 28 mol%, about 29 mol%, about 30 mol%, about 35 mol%, about 40 mol%, about 45 mol%, about 50 mol%, about 55 mol%, about 60 mol%, about 65 mol%, about 70 mol%, about 75 mol%, about 80 mol%, about 85 mol%, about 90 mol%, about 95 mol% to about 100 mol% cholesterol (i.e., a 1:1 mol / mol mixture of cholesterol and phospholipids (e.g., DMPC)). In an embodiment, the HDL-derived nanoparticles contain about 1 mol% to about 30 mol% cholesterol. In an embodiment, the HDL-derived nanoparticles contain about 15 mol% to about 25 mol% cholesterol relative to the phospholipid. In an embodiment, the HDL-derived nanoparticles contain about 20 mol% cholesterol relative to the phospholipid. In an embodiment, the HDL-derived nanoparticles contain about 10 mol% to about 35 mol% cholesterol relative to the phospholipid. In an embodiment, the HDL-derived nanoparticles contain about 15 mol% to about 30 mol% cholesterol relative to the phospholipid. In an embodiment, the HDL-derived nanoparticles contain about 15 mol% to about 25 mol% cholesterol relative to the phospholipid. In an embodiment, the HDL-derived nanoparticles contain about 28 mol% to about 23 mol% cholesterol relative to the phospholipid. In an embodiment, the HDL-derived nanoparticles contain about 20 mol% to about 27 mol% cholesterol relative to the phospholipid.

[0227] In embodiments, the HDL-derived nanoparticles do not contain cholesterol. In embodiments, the molar ratio of cholesterol:phospholipids in the HDL-derived nanoparticles is about 0.025:1, about 0.05:1, about 0.075:1, about 0.1:1, about 0.125:1, about 0.15:1, about 0.175:1, about 0.2:1, about 0.225:1, about 0.25:1, about 0.275:1, about 0.3:1, about 0.325:1, about 0.35:1, about 0.375:1, about 0.4:1, about 0.425:1, about 0.45:1, about 0.475:1 or about 0.5:1, including all values ​​therebetween. The molar ratio of cholesterol:phospholipids ranges from about 0:1 to about 0.5:1, for example, about 0:1, about 0.025:1, about 0.05:1, about 0.075:1, about 0.1:1, about 0.125:1, about 0.15:1, about 0.175:1, about 0.2:1, about 0.225:1, about 0.25:1, about 0.275:1, about 0.3:1, about 0.325:1, about 0.35:1, about 0.375:1, about 0.4:1, about 0.425:1, about 0.45:1, about 0.475:1 to about 0.5:1, including all ranges therebetween. In an embodiment, the molar ratio of cholesterol:phospholipids ranges from about 0.05:1 to about 0.25:1. In an embodiment, the molar ratio of cholesterol is about 0.2:1.

[0228] In embodiments, the HDL-derived nanoparticles comprise one or more phospholipids and cholesterol in a molar ratio ranging from about 1:0.05 to about 1:0.25, In embodiments, the HDL-derived nanoparticles comprise one or more phospholipids and cholesterol in a molar ratio of 1:0.2.

[0229] In embodiments, the weight percentage of cholesterol ranges from about 0% (w / w) to about 15% (w / w) of the nanoparticle, lipid, or composition, e.g., about 1% (w / w), about 1.5% (w / w), about 2% (w / w), about 2.5% (w / w), about 3% (w / w), about 3.5% (w / w), about 4% (w / w), about 4.5% (w / w), about 5% (w / w), about 5.5% (w / w), about 6% (w / w), about 6.5% (w / w). ), about 7% (w / w), about 7.5% (w / w), about 8% (w / w), about 8.5% (w / w), about 9% (w / w), about 9.5% (w / w), about 10% (w / w), about 10.5% (w / w), about 11% (w / w), about 11.5% (w / w), about 12% (w / w), about 12.5% ​​(w / w), about 13% (w / w), about 13.5% (w / w), about 14% (w / w), about 14.5% (w / w) to about 15% (w / w). In embodiments, the weight percentage of cholesterol ranges from about 0% (w / w) to about 15% (w / w) of the nanoparticle, lipid, or composition, e.g., about 1% (w / w), about 1.5% (w / w), about 2% (w / w), about 2.5% (w / w), about 3% (w / w), about 3.5% (w / w), about 4% (w / w), about 4.5% (w / w), about 5% (w / w), about 5.5% (w / w), about 6% (w / w), about 6.5% (w / w). ), about 7% (w / w), about 7.5% (w / w), about 8% (w / w), about 8.5% (w / w), about 9% (w / w), about 9.5% (w / w), about 10% (w / w), about 10.5% (w / w), about 11% (w / w), about 11.5% (w / w), about 12% (w / w), about 12.5% ​​(w / w), about 13% (w / w), about 13.5% (w / w), about 14% (w / w), about 14.5% (w / w) to about 15% (w / w). In an embodiment, the weight percentage is the weight percentage of cholesterol relative to the phospholipid. In an embodiment, the weight percentage of cholesterol is in the range of about 1-10% cholesterol (w / w%) of the composition. The weight percentage of cholesterol ranges from about 2-8% cholesterol (w / w%) of the composition. In embodiments, the weight percentage of cholesterol ranges from about 3.5-7.5% cholesterol (w / w%) of the composition.In embodiments, the weight percentage of cholesterol ranges from about 5-10% cholesterol (w / w%) of the composition. In embodiments, the weight percentage of cholesterol is about 3.6 (w / w%) of the composition. In embodiments, the weight percentage of cholesterol is about 7.2 (w / w%) of the composition. In embodiments, the weight percentage of cholesterol is about 5.9 (w / w%) of the composition.

[0230] In embodiments, the size and circulation time of nanoparticles can be modulated, for example, by controlling the ratio of lipid to APOA1, and the ratio of lipid to polymer or lipid to triglyceride.

[0231] In embodiments, the HDL-derived nanoparticles are in a ratio (e.g., on a molar basis) of about 5:1 to 1000:1, e.g., about 5:1, about 10:1, about 20:1, about 30:1, about 40:1, about 50:1, about 60:1, about 70:1, about 80:1, about 90:1, about 100:1, about 110:1, about 120:1, about 130:1, about 140:1, about 150:1, about 160:1, about 170:1, about 180:1, about 190:1, about 200:1, about 210:1, about 220:1 About 230:1, about 240:1, about 250:1, about 260:1, about 270:1, about 280:1, about 290:1, about 300:1, about 310:1, about 320:1, about 330:1, about 340:1, about 350:1, about 360:1, about 370:1, about 380:1, about 390:1, about 400:1, about 410:1, about 420:1, about 430:1, about 440:1, about 450:1, about 460:1, about 470:1 , about 480:1, about 490:1, about 500:1, about 510:1, about 520:1, about 530:1, about 540:1, about 550:1, about 560:1, about 570:1, about 580:1, about 590:1, about 600:1, about 610:1, about 620:1, about 630:1, about 640:1, about 650:1, about 660:1, about 670:1, about 680:1, about 690:1, about 700:1, about 710:1, about 720:1 phospholipid:apoA-I or apoA-I mimetic of about 730:1, about 740:1, about 750:1, about 760:1, about 770:1, about 780:1, about 790:1, about 800:1, about 810:1, about 820:1, about 830:1, about 840:1, about 850:1, about 860:1, about 870:1, about 880:1, about 890:1, about 900:1, about 910:1, about 920:1, about 930:1, about 940:1, about 950:1, about 960:1, about 970:1, about 980:1, about 990:1 to about 1000:1, including all subranges and values ​​therebetween. In embodiments, the HDL-derived nanoparticles comprise a ratio (e.g., on a molar basis) of phospholipids:apoA-I or apoA-I mimetic of about 10:1 to 1000:1. In embodiments, the HDL-derived nanoparticles comprise a ratio (e.g., on a molar basis) of phospholipids:apoA-I of about 70:1 to 125:1. In embodiments, the HDL-derived nanoparticles comprise a ratio (e.g., on a molar basis) of apoA-I mimetic of about 5:1 to 10:1.

[0232] In embodiments, the HDL-derived nanoparticles comprise a weight ratio of about 2:1 to 3:1 phospholipid:apoA-I or apoA-I mimetic.

[0233] In embodiments, the HDL-derived nanoparticles comprise about or at least about 0.1 mol% to about 100 mol% of a compound of formula I relative to the phospholipid (e.g., DMPC), e.g., about or at least about 0.1 mol%, about or at least about 0.5 mol%, about or at least about 0.75 mol%, about or at least about 1% mol%, about or at least about 2 mol%, about or at least about 3 mol%, about or at least about 4 mol%, about or at least about 5 mol%, about or at least about 6 mol%, about or at least about 7 mol%, about or at least about 8 mol%, about or at least about 9 mol%, about or at least about 10 mol%, about or at least about 12 mol%, about or at least about 14 mol%, about or at least about 15 mol%, about or at least about 16 mol%, about or at least about 17 mol%, about or at least about 18 mol%, about or at least about 19 mol%, about or at least about 20 mol%, about or at least about 22 mol%, about or at least about 24 mol%, about or at least about 25 mol%, about or at least about 26 mol%, about or at least about 28 mol%, about or at least about 29 mol%, about or at least about 30 mol%, about or at least about 31 mol%, about or at least about 32 mol%, about or at least about 33 mol%, about or at least about 34 mol%, about or at least about 35 mol%, about or at least about 36 mol%, about or at least about 37 mol%, about or at least about 38 mol%, about or at least about 39 mol%, about or at least about 40 mol%, about or at least about 42 mol%, about or at least about 44 mol%, about or at least about 45 mol%, about or at least about 7 mol%, about or at least about 8 mol%, about or at least about 9 mol%, about or at least about 10 mol%, about or at least about 11 mol%, about or at least about 12 mol%, about or at least about 13 mol%, about or at least about 14 mol%, about or at least about 15 mol%, about or at least about 16 mol%, about or at least about 17 mol%, about or at least about 18 mol%, about or at least about 19 mol%, about or at least about 20 mol%, about or is at least about 21 mol%, about or at least about 22 mol%, about or at least about 23 mol%, about or at least about 24 mol%, about or at least about 25 mol%, about or at least about 26 mol%, about or at least about 27 mol%, about or at least about 28 mol%, about or at least about 29 mol%, to about or at least about 30 mol%, about or at least about 35 mol%, about or at least about 40 mol%, about or at least about 45 mol%, about or at least about 50 mol%, %, about or at least about 55 mol%, about or at least about 60 mol%, about or at least about 65 mol%, about or at least about 70 mol%, about or at least about 75 mol%, about or at least about 80 mol%, about or at least about 85 mol%, about or at least about 90 mol%, about or at least about 95 mol% to about or at least about 100 mol% of the compound of formula I (a 1:1 mol / mol mixture of the compound and phospholipid (e.g., DMPC)), including all ranges and values ​​therebetween. In embodiments, the HDL-derived nanoparticles comprise about 10 mol% to about 30 mol% of the compound of formula I relative to the phospholipid.In an embodiment, the HDL-derived nanoparticles contain about 12 mol % to about 25 mol % of the compound relative to the phospholipid.

[0234] In embodiments, the nanoparticle size ranges from about 5 nm to about 500 nm in diameter, e.g., about 5 nm, about 10 nm, about 15 nm, about 20 nm, about 25 nm, about 30 nm, about 35 nm, about 40 nm, about 50 nm, about 55 nm, about 60 nm, about 65 nm, about 70 nm, about 75 nm, about 80 nm, about 85 nm, about 90 nm, about 95 nm to about 100 nm, about 110 nm, about 120 nm, about 130 nm, about 140 nm, about 150 nm. The nanoparticle size ranges from about 160 nm, about 170 nm, about 180 nm, about 190 nm, about 200 nm, about 210 nm, about 220 nm, about 230 nm, about 240 nm, about 250 nm, about 260 nm, about 270 nm, about 280 nm, about 290 nm, about 300 nm, about 310 nm, about 320 nm, about 330 nm, about 340 nm, about 350 nm, about 360 nm, about 370 nm, about 380 nm, about 390 nm, about 400 nm, about 410 nm, about 420 nm, about 430 nm, about 440 nm, about 450 nm, about 460 nm, about 470 nm, about 480 nm, about 490 nm to about 500 nm, including all ranges and values ​​therebetween. In embodiments, the nanoparticle size is less than about 50 nm. In embodiments, the nanoparticle size is about 50 nm to about 100 nm, or about 5 nm to about 30 nm. In embodiments, the nanoparticle size is measured by dynamic light scattering (DLS). In embodiments, nanoparticles with long blood half-lives and small size (<50 nm) can be used to target immune cells in tissues with limited access to the circulation. In embodiments, nanoparticles with short blood half-lives and large size (about 100 nm) can be used to target immune cells in well-perfused tissues. These tissues include the spleen, liver, kidney, lung, and bone marrow.

[0235] In embodiments, the HDL-derived nanoparticles are disc-shaped. In embodiments, the HDL-derived nanoparticles are spherical. In embodiments, the morphology of the HDL-derived nanoparticles is visualized by transmission electron microscopy (TEM).

[0236] In embodiments, the HDL-derived nanoparticles are about 5 to about 100 nm in length, e.g., about 5 nm, about 10 nm, about 15 nm, about 20 nm, about 25 nm, about 30 nm, about 35 nm, about 40 nm, about 50 nm, about 55 nm, about 60 nm, about 65 nm, about 70 nm, about 75 nm, about 80 nm, about 85 nm, about 90 nm, about 95 nm to about 100 nm in length, including all ranges and values ​​therebetween. In embodiments, the HDL-derived nanoparticles are about 10 nm to 80 nm in length. In embodiments, the HDL-derived nanoparticles are about 15 nm to 50 nm in length. In embodiments, the HDL-derived nanoparticles are greater than about 10 nm, or greater than about 15 nm. In embodiments, the HDL-derived nanoparticles have a thickness of about 1 nm to 10 nm, e.g., about 1 nm, about 2 nm, about 3 nm, about 4 nm, about 5 nm, about 6 nm, about 7 nm, about 8 nm, about 9 nm to about 10 nm, including all ranges and values ​​therebetween. In embodiments, the thickness of the HDL particles is about 1 to 10 nm, or 2 to 7 nm, or 3 to 6 nm. In embodiments, the dimensions (e.g., length and thickness) are recorded by cryo-TEM. In embodiments, the HDL particles have a worm-like morphology by cryo-TEM.

[0237] In embodiments, the HDL-derived nanoparticles are disk-shaped with diameters of about 5 nm to about 50 nm (e.g., as measured by dynamic light scattering (DLS)), e.g., about 5 nm, about 10 nm, about 15 nm, about 20 nm, about 25 nm, about 30 nm, about 40 nm to about 50 nm, including all subranges and values ​​therebetween. In embodiments, the nanodiscs are about 5 nm to about 30 nm in diameter.

[0238] In embodiments, the HDL-derived nanoparticles are spherical with a diameter of about 10 nm to about 400 nm (e.g., as measured by dynamic light scattering (DLS)), e.g., diameters of about 10 nm, about 15 nm, about 20 nm, about 30 nm, about 40 nm, about 50 nm, about 60 nm, about 70 nm, about 80 nm, about 100 nm, about 110 nm, about 120 nm, about 130 nm, about 140 nm, about 150 nm, about 160 nm, about 170 nm, about 180 nm, about 190 nm, about 210 nm, about 220 nm, about 230 nm, about 240 nm, about 250 nm, about 260 nm, about 270 nm, about 280 nm, about 290 nm, about 300 nm, about 310 nm, about 320 nm, about 330 nm, about 340 nm, about 350 nm, about 360 nm, about 370 nm, about 380 nm, about 390 nm, about 400 nm, about 410 nm, about 420 nm, about 430 nm, about 440 nm, about 450 nm, about 460 nm, about 470 nm, about 480 nm, about 490 nm, about 500 nm, about 510 nm, about 520 nm, about 530 nm, about 540 nm, about 550 nm, about 560 nm, about 570 nm, about 580 nm, about 590 nm, about 600 nm, about 600 nm, about 610 nm, about 620 nm, about 630 nm, about 640 nm, about 650 nm, about 660 nm, about 6 m, about 180 nm, about 190 nm, about 200 nm, about 210 nm, about 220 nm, about 230 nm, about 240 nm, about 250 nm, about 260 nm, about 270 nm, about 280 nm, about 290 nm, about 300 nm, about 310 nm, about 330 nm, about 340 nm, about 350 nm, about 360 nm, about 370 nm, about 380 nm, about 390 nm to about 400 nm, including all values ​​and ranges therebetween. In embodiments, the nanospheres are between about 15 nm and about 250 nm in diameter. In embodiments, the nanospheres are between about 30 nm and about 100 nm in diameter.

[0239] The stability of the HDL-derived nanoparticles can be assessed by performing DLS measurements. In embodiments, the HDL-derived nanoparticles are stable, for example by DLS, for at least about 1 week, or at least about 2 weeks, or at least about 5 weeks.

[0240] In embodiments, the nanobiological composition promotes a hyper-reactive innate immune response in a patient in need thereof. In embodiments, the hyper-reactive innate immune response is promoted for at least about 7 to about 30 days. In embodiments, the hyper-reactive innate immune response is promoted for at least 30 to 100 days. In embodiments, the hyper-reactive innate immune response is promoted for more than 100 days and up to 3 years. In embodiments, the nanobiological composition is administered once and the hyper-reactive innate immune response is promoted for at least 30 days. In embodiments, the nanobiological composition is administered at least once daily on each day of a multiple dose regimen and the hyper-reactive innate immune response is promoted for at least 30 days.

[0241] The production method can prepare either uniformly sized HDL-derived nanoparticles or a mixture of heterogeneous sized HDL-derived nanoparticles, either by not filtering or by preparing HDL-derived nanoparticles of various sizes and recombining them in a post-production step. The larger the size of the HDL-derived nanoparticles, the more drug they can incorporate. However, larger sizes, e.g., >120 nm, may limit, impede, or delay the diffusion of HDL-derived nanoparticles into the tissues of the patient undergoing treatment. Smaller HDL-derived nanoparticles do not carry as much drug per particle, but can access bone marrow cells, bone marrow progenitor cells, and hematopoietic stem cells (biodistribution), such as bone marrow, blood, or spleen, or other local tissues that are subject to trained immunity, e.g., bone marrow, blood, and / or spleen.

[0242] Using a heterogeneous mixture of nanoparticle sizes in a single dose or regimen, it is possible to immediately reduce innate immune hyperresponsiveness, as well as provide durable and long-term relief of innate immune hyperresponsiveness that can last for days, weeks, months, and years, and the nanobiologics are reversing, modifying, or re-regulating metabolic, epigenetic, and inflammasome pathways in hematopoietic stem cells (HSCs), common myeloid progenitor cells (CMPs), and myeloid cells, e.g., monocytes, macrophages, and other short-lived circulating cells.

[0243] In embodiments, the maximum loading capacity of HDL-derived nanoparticles can be determined by dividing the internal volume of the HDL-derived nanoparticles by the volume of the drug-loaded spheroids. Particles: We assume 100 nm spherical particles with phospholipid walls of 2.2 nm to 3.0 nm, with an inner diameter of 94 nm and a volume (large) @ 4 / 3n(r)3. Consider the drug:stimulator as a 12x12x35 angstrom, or 1.2x1.2x3.5 nm cylinder, and the multi-drug molecule cylinder as a 3.5 nm diameter spheroid with radius 1.75 nm and volume(small)@4 / 3n(r), e.g. 7 or 9. Maximum packing capacity (calculated value): Approximately 487k for 3.5 nm spheroids within a 100 nm particle.

[0244] formulation When used as medicine, the compound of the present disclosure and the HDL-derived nanoparticles are typically administered in the form of pharmaceutical composition.Such composition can be prepared by a method well known in the pharmaceutical field and comprises at least one active compound.In an embodiment, the pharmaceutical composition comprises the nanobiological composition of the present disclosure and a pharmaceutically acceptable carrier.

[0245] In general, the compounds of the present invention are administered in a pharma- ceutically effective amount. The amount of compound actually administered will typically be determined by the physician, taking into account the relevant circumstances, including the condition to be treated, the selected route of administration, the actual compound to be administered, the age, weight, and response of the individual patient, the severity of the patient's symptoms, etc.

[0246] The pharmaceutical composition of the present invention can be administered by various routes, including oral, rectal, intraocular, transdermal, subcutaneous, intravenous, intramuscular, intraperitoneal, intradermal, directly into cerebrospinal fluid, intratracheal, and intranasal.Depending on the intended delivery route, the compound of the present invention is preferably formulated as either an injectable or oral composition, or as a salve, lotion, or patch, all for transdermal administration.In an embodiment, the composition is administered intravenously or intraarterially.

[0247] Compositions for oral administration can take the form of bulk liquid solutions or suspensions, or bulk powders. More commonly, however, compositions are provided in unit dosage forms to facilitate accurate dosing. The term "unit dosage form" refers to physically discrete units suitable as unitary dosages for human subjects and other mammals, each unit containing a predetermined amount of active material calculated to produce a desired therapeutic effect in association with suitable pharmaceutical excipients. Typical unit dosage forms include prefilled, premeasured ampoules or syringes of liquid compositions, or pills, tablets, capsules, and the like in the case of solid compositions. In such compositions, the compounds described herein are usually minor components (about 0.1 to about 50% by weight, or preferably about 1 to about 40% by weight), with the remainder being various vehicles or carriers and processing aids that serve to form the desired dosage form.

[0248] Liquid forms suitable for oral administration may include a suitable aqueous or non-aqueous vehicle, including buffers, suspending and dispersing agents, colorants, flavorings, etc. Solid forms may include, for example, any of the following ingredients: binders such as microcrystalline cellulose, gum tragacanth, or gelatin; excipients such as starch or lactose, disintegrating agents such as alginic acid, Primogel, or corn starch; lubricants such as magnesium stearate; glidants such as colloidal silicon dioxide; sweeteners such as sucrose or saccharin; or flavoring agents such as peppermint, methyl salicylate, or orange flavoring, or compounds of a similar nature.

[0249] Injectable compositions are typically based on injectable sterile saline, phosphate buffered saline, or other injectable carriers known in the art. As mentioned above, the active compound in such compositions is typically a minor component, often about 0.05-10% by weight, with the remainder being the injectable carrier, etc.

[0250] Transdermal compositions are typically formulated as topical ointments or creams containing the active ingredient, generally in an amount ranging from about 0.01 to about 20% by weight, preferably about 0.1 to about 20% by weight, preferably about 0.1 to about 10% by weight, more preferably about 0.5 to about 15% by weight. When formulated as an ointment, the active ingredient is typically combined with either a paraffinic or water-miscible ointment base. Alternatively, the active ingredient may be formulated in a cream, for example, with an oil-in-water cream base. Such transdermal formulations are well known in the art and generally include additional ingredients to enhance the stability of the active ingredient or formulation for skin penetration. All such known transdermal formulations and ingredients are included within the scope of the present invention.

[0251] The nanoparticles described herein can also be administered by a transdermal device.Thus, transdermal administration can be accomplished using a patch either of the reservoir or porous membrane type, or of a solid matrix variety.

[0252] The above ingredients for compositions for oral administration, injection or topical administration are merely representative. Other materials and processing techniques, etc., are described in Remington's Pharmaceutical Sciences, 17 th edition, 1985, Mack Publishing Company, Easton, Pennsylvania, Part 8, which is incorporated herein by reference.

[0253] For injection, the nanoparticles described herein can be provided in injectable saline solutions, in the form of injectable liposomal solutions, slow release polymer systems, and the like.

[0254] The nanoparticles described herein can also be administered in sustained release forms or from sustained release drug delivery systems. A description of representative sustained release materials can be found in Remington's Pharmaceutical Sciences.

[0255] method For example, provided herein are methods of treating subjects susceptible to or suffering from immune-related diseases and conditions, including immune paralysis in sepsis and infection, cell proliferation disorders (such as cancer), and other diseases and conditions caused by defects in trained immunity.

[0256] In embodiments, the present disclosure provides a method for treating a cell proliferation disorder, comprising administering to a subject in need thereof a therapeutically effective amount of a nanobiological composition comprising high density lipoprotein (HDL)-derived nanoparticles comprising a compound of the present disclosure, such as a compound of formula I. In embodiments, the compounds, compositions provided herein are useful for treating cancer by inducing trained immunity.

[0257] In embodiments, the cell proliferation disorder is cancer. In embodiments, the cancer is one or more of the following cancers: aggressive malignant tumors, amyloidosis, neuroblastoma, meningioma, hemangiopericytoma, multiple brain metastases, glioblastoma multiforme, glioblastoma, brain stem glioma, poor prognosis malignant brain tumors, malignant glioma, recurrent malignant glioma, anaplastic astrocytoma, anaplastic oligodendroglioma, neuroendocrine tumors, rectal adenocarcinoma, Dukes C and D colorectal cancer, unresectable colorectal cancer, metastatic hepatocellular carcinoma, Kaposi's sarcoma, karyotype acute myeloblastic leukemia, Hodgkin's lymphoma, non-Hodgkin's lymphoma, cutaneous T-cell lymphoma, cutaneous B-cell lymphoma, diffuse large B-cell lymphoma, low-grade follicular lymphoma, and / or cytoplasmic lymphoma. tumor, malignant melanoma, malignant mesothelioma, malignant pleural effusion mesothelioma syndrome, peritoneal carcinoma, papillary serous adenocarcinoma, gynecological sarcoma, soft tissue sarcoma, scleroderma, cutaneous vasculitis, Langerhans cell histiocytosis, leiomyosarcoma, fibrodysplasia ossificans progressiva, hormone refractory prostate cancer, resected high-risk soft tissue sarcoma, unresectable hepatocellular carcinoma, Waldenström's macroglobulinemia, smoldering myeloma, indolent myeloma, fallopian tube cancer, androgen-independent prostate cancer, androgen-dependent stage IV non-metastatic prostate cancer, hormone-insensitive prostate cancer, chemotherapy-insensitive prostate cancer, papillary thyroid cancer, follicular thyroid cancer, medullary thyroid cancer, and leiomyoma. In an embodiment, the cancer is selected from the group consisting of bladder cancer, vascular cancer, bone cancer, brain cancer, breast cancer, cervical cancer, breast cancer, colon cancer, endometrial cancer, esophageal cancer, eye cancer, head cancer, kidney cancer, liver cancer, lymph node cancer, lung cancer, oral cancer, cervical cancer, ovarian cancer, pancreatic cancer, prostate cancer, rectal cancer, colorectal cancer, skin cancer, stomach cancer, testicular cancer, pharyngeal cancer, thyroid cancer, urothelial cancer, and uterine cancer. In an embodiment, the cancer is selected from the group consisting of breast cancer, prostate cancer, melanoma, colorectal cancer, lung cancer, pancreatic cancer, and glioblastoma. In an embodiment, the cancer is metastatic. In an embodiment, the cancer is refractory or resistant to chemotherapy or radiation, particularly refractory to thalidomide.

[0258] In embodiments, the cancer is selected from the group consisting of bladder cancer, vascular cancer, bone cancer, brain cancer, breast cancer, cervical cancer, breast cancer, colon cancer, endometrial cancer, esophageal cancer, eye cancer, head cancer, kidney cancer, liver cancer, lymph node cancer, lung cancer, oral cancer, cervical cancer, ovarian cancer, pancreatic cancer, prostate cancer, rectal cancer, colorectal cancer, skin cancer, stomach cancer, testicular cancer, pharyngeal cancer, thyroid cancer, urothelial cancer, and uterine cancer.

[0259] In embodiments, the cancer is selected from the group consisting of breast cancer, prostate cancer, melanoma, colorectal cancer, lung cancer, pancreatic cancer, and glioblastoma.

[0260] In embodiments, the present disclosure provides a method for treating sepsis comprising administering to a subject in need thereof a therapeutically effective amount of a nanobiological composition of the present disclosure, in embodiments, the patient has sepsis associated with a bacterial, viral or fungal infection of the lungs, abdomen, kidneys, or bloodstream.

[0261] The compounds of the present disclosure and their carriers disclosed herein can be used to enhance immune response.Therefore, disclosed herein is a method of inducing immune response, comprising administering to a subject an immunogenic composition, the composition comprising (i) at least one antigen and (ii) the compounds disclosed herein, optionally in a nanoparticle carrier, such as HDL-derived nanoparticles or liposomes.

[0262] The antigen is typically derived from a pathogen, but neo-antigens from a subject with cancer can also be used. Exemplary pathogen antigens can be derived from viruses, bacteria, parasites, or yeast. In an embodiment, the antigen can be secreted from a pathogen; for example, it can be an exotoxin or an endotoxin.

[0263] Examples of viruses include adenovirus, adeno-associated virus (AAV), chikungunya, dengue, influenza, Ebola, Epstein-Barr, Hanta, hepatitis (e.g., Hepatitis A, B, C, D, E), CMV, HPV (e.g., one or more of HPV1-18), coronavirus (e.g., SARS, MERS, COVID-19), polio, rabies, and Zika. Examples of bacteria include Vibrio cholerae, E. coli, Salmonella spp., N. gonorrhea, N. meningitidis, Streptococcus pyogenes, Mycobacterium tuberculosis, Legionella pneumophila, Brucella bortus, and Listeria monocytogenes.

[0264] The antigen may be, for example, a polypeptide, including a glycosylated peptide, or a carbohydrate. In an embodiment, the immunogenic composition may include a nucleic acid encoding an antigen, typically a polypeptide that is transcribed and / or translated from a nucleic acid. The nucleic acid may be DNA or RNA, or may be a derivative of DNA or RNA. Common RNA derivatives include covalent modifications to the molecule to enhance stability and / or expression. In an embodiment, the nucleic acid encoding the polypeptide may be in a plasmid or a viral vector, such as an adenovirus vector, an adeno-associated virus vector, a baculovirus vector, or a lentivirus vector.

[0265] In aspects, the administration may be prophylactic; for example, vaccinating a subject prior to exposure to a pathogen. In other aspects, the administration may be therapeutic; for example, inducing an immune response against neoantigen-bearing tumors in a subject suffering from cancer. In embodiments, the nanobiological composition is administered to a patient in a treatment regimen comprising two or more doses to generate accumulation of the drug in bone marrow cells, bone marrow progenitor cells, and hematopoietic stem cells in the bone marrow, blood, and / or spleen.

[0266] In embodiments, the nanobiological composition is administered intravenously or intraarterially.

[0267] Injection dose levels range from about 0.1 mg / kg / hour to at least 10 mg / kg / hour, all for about 1 to about 120 hours, particularly 24 to 96 hours. A preloading bolus of about 0.1 mg / kg to about 10 mg / kg or more may also be administered to achieve adequate steady state levels. The maximum total dose is not expected to exceed about 2 g / day for a human patient weighing 40 to 80 kg.

[0268] Oral dosage levels range from about 0.01 to about 20 mg / kg of a compound of the invention, including all ranges and values ​​therebetween, for example, dosage levels range from about 0.1 to about 10 mg / kg, or from about 1 to about 5 mg / kg.

[0269] Transdermal doses are generally selected to provide blood levels that are comparable to or lower than those achieved using injection doses.Appropriate modes of administration for mucosal sites are also contemplated herein, including but not limited to anal swabs, enemas, nasal sprays, and aerosolized or vaporized compounds and / or compositions for delivery to pulmonary mucosa.Those skilled in the art will select appropriate delivery models based on a variety of parameters, including the organ or tissue site of the patient with the disease or condition that is most severely affected by the disease or condition.

[0270] The compounds of the present invention can be administered as the sole active agent, or can be administered in combination with one or more additional agents, including other compounds that exhibit the same or similar therapeutic activity, and such combined administration is determined to be safe and effective.In an embodiment, the additional agent is an inhibitor of checkpoint protein.In an embodiment, the method provided herein further comprises co-administering an anti-cancer drug with the nanobiological composition as a combination therapy.

[0271] A compound or composition described herein can be provided in a kit. In some embodiments, the kit includes (a) a compound described herein, or a composition comprising a compound described herein (where, for example, the compound can be a NOD2 modulator described herein), and, optionally, (b) informational material. The informational material can be descriptive, instructional, marketing, or other material related to the methods described herein and / or the use of the compounds or compositions described herein for the methods described herein. In embodiments, the informational material can include information regarding the production of the compound. In embodiments, the informational material relates to methods of administering the compound. In embodiments, the informational material can include instructions for administering a compound or composition described herein in a suitable manner, e.g., in a suitable dose, dosage form, or mode of administration (e.g., a dose, dosage form, or mode of administration described herein), for carrying out the methods described herein. In embodiments, the informational material can include instructions for administering a compound described herein to a suitable subject, e.g., a human, e.g., a human having or at risk for a disorder described herein.

[0272] The kit may include one or more containers for the composition containing the compound or composition described herein. In embodiments, the kit contains separate containers, dividers or compartments for the composition and the informational material. For example, the composition may be contained in a bottle, vial, or syringe, and the informational material may be contained in a plastic sleeve or packet. In embodiments, the separate elements of the kit are contained within a single undivided container. For example, the composition is contained in a bottle, vial, or syringe to which the informational material in the form of a label is attached. In embodiments, the kit includes a plurality (e.g., pack) of individual containers, each of which contains one or more unit dosage forms (e.g., dosage forms described herein) of the compounds or compositions described herein. For example, the kit includes a plurality of syringes, ampoules, foil packets, or blister packs, each of which contains a single unit dose of the compounds described herein. The containers of the kit may be airtight, waterproof (e.g., impermeable to changes in moisture or evaporation), and / or light-tight.

[0273] Also provided herein is a process for producing a nanobiological composition of the present disclosure, the process comprising: a) forming, under conditions effective to form a lipid membrane, a lipid membrane comprising: i) a compound of the present disclosure; ii) one or more phospholipids; optionally, iii) a hydrophobic matrix comprising one or more triglycerides, fatty acid esters, hydrophobic polymers, or sterol esters, or combinations thereof; and optionally iv) cholesterol; and b) dissolving the lipid membrane in a solvent to form a lipid solution; and contacting the lipid solution with apoA-I or a peptidomimetic of apoA-I under conditions effective to form HDL-derived nanoparticles comprising a compound of the present disclosure.

[0274] In embodiments, provided herein are nanobiological compositions prepared according to the methods disclosed herein. EXAMPLES

[0275] The therapeutic agents described herein and nanoparticles containing same can be prepared from known or commercially available starting materials and reagents by those skilled in the art of organic synthesis.

[0276] material and method All chemicals were purchased from commercial sources and used without further purification unless otherwise specified. N-Methylmorpholine was redistilled and the fraction between 110 °C and 112 °C was collected. Cholesterol azidoacetate was synthesized according to known procedures (RSC Adv. 2015, 5, 12094), as was 1-azidooctadecane. Dry solvents were obtained using an MBRAUN solvent purification system (MB-SPS). Toluene was dried over 4 Å molecular sieves before use. Glassware used for reactions carried out under an argon atmosphere was dried with a heat gun before use. Thin layer chromatography (TLC) was performed using Merck 60-F254 silica gel plates and visualized by 254 nm UV light, permanganate staining and / or cerium molybdate (CeMo) staining. Normal and reverse phase automated column chromatography was performed on a Biotage Isolera One or Grace Reveleris X2 flash chromatography system using Biotage Sfar Silica, Buchi FlashPure ID Silica, or Buchi FlashPure ID C18 columns. Elution gradients are specified in column volumes (CV). Unstabilized THF was used for the water / THF gradient.

[0277] NMR spectra were recorded on a Bruker 400 MHz Ultrashield spectrometer (400 MHz for 1H NMR). The deuterated solvent used is indicated in each case. Chemical shifts (δ) are expressed in ppm and refer to residual peaks of the solvent. Peak multiplicities are abbreviated as follows: s: singlet; d: doublet; t: triplet; dt: doublet of triplets; ddt: doublet of triplet doublets; td: triplet of doublets; tt: triplet of triplets; q: quartet; ABq: AB quartet; dq: doublet of quartets; qd: quartet of doublets; sept: septet; m: multiplet; bs: broad singlet. Matrix-assisted laser desorption / ionization time-of-flight (MALDI-TOF) mass spectra were acquired on a PerSeptive Biosystems Voyager DE-PRO spectrometer using α-cyano-4-hydroxycinnamic acid (CHCA) or trans-2-[3-(4-tert-butylphenyl)-2-methyl-2-propenylidene]-malononitrile (DCTB) as matrices. Gas chromatography-mass spectrometry (GC-MS) measurements were performed on a Shimadzu GC-17A gas chromatograph equipped with a Shimadzu AOC-20i autoinjector, a Shimadzu GCMS-QP5000 gas chromatograph-mass spectrometer, and a Phenomenex Zebron ZB-35 column (l = 30 meters, ID = 0.25 mm, film thickness = 0.25 μm). High performance liquid chromatography mass spectrometry (HPLC-ESI-MS) experiments using a water / acetonitrile gradient were performed on a Shimadzu setup equipped with 2xLC-20AD pumps, DGU-20A3 degasser, SIL-20AC autosampler, SPD-M20A PDA, and a ThermoScientific LCQ fleet MS. Column: Phenomenex Kinetex 5um EVO C18 100Å LC (50×2.1mm). Gradient: water / MeCN (+0.1% formic acid) 5–100% MeCN, 0.300mL / min. Electrospray ionization (ESI) was used to generate charge for MS detection.HPLC-MS and HPLC-ELSD experiments using water / THF or water / MeOH gradients were performed on a Shimadzu Nexera-iLC-2040C 3D Plus equipped with a Shimadzu LCMS-8045. Column: Alltech Alltima C18 (150×3.2 mm; 5 um; no. 88383). Gradient: water / THF (+0.1% TFA) or water / MeOH (+0.1% TFA), 0.400 mL / min. This HPLC setup was used in combination with ELSD (evaporative light scattering detection).

[0278] Alternatively, HPLC-MS (SIM) and HPLC-ELSD were performed on a Phenomenex Kinetex 5 micrometer EVO C18 100A LC column (50 × 2.1 mm) using a gradient from eluent A to B, A = 20 mM NH4HCO2 in H2O containing 0.1 v / v % formic acid, and B = 2-propanol / MeCN / H2O 85:15:5, also containing 20 mM NH4HCO2 and 0.1 v / v % formic acid.

[0279] Abbreviation HPLC = high performance liquid chromatography; ELSD = evaporative light scattering detection; ESI-MS = electrospray ionization mass spectrometry; SIM = selected ion mode; NMR = nuclear magnetic resonance.

[0280] [ka]

[0281] MDP = Muramyl dipeptide muramyl (or N-acetylmuramyl-L-alanyl-D-isoglutamine) CAS [53678-77-6]. Prepared according to standard peptide synthesis or purchased from commercial sources.

[0282] NHS = N-hydroxysuccinimide; DiC or DIC = N,N'-diisopropylcarbodiimide; EDC = N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide (use the hydrochloride salt); PyBOP = (benzotriazol-1-yloxytripyrrolidinophosphonium hexafluorophosphate); SPPS = solid phase peptide synthesis.

[0283] TEA = triethylamine; THF = tetrahydrofuran; MeOH = methanol; DMF = dimethylformamide; FA = formic acid; TFA = trifluoroacetic acid.

[0284] Building Blocks DSPE-Azide Acetate (DSPE-CO-CH2-N3)

[0285] [ka] (2R)-3-(((2-aminoethoxy)(hydroxy)phosphoryl)oxy)propane-1,2-distearate (260 mg, 0.35 mmol), 2,3,5,6-tetrafluorophenyl 2-azidoacetate (prepared by D. J. Vugts et al., Bioconjugate Chem. 2011, 22, 2072-2081; 87 mg, 0.35 mmol, 1 equiv.), and N,N-diisopropylethylamine (184 μL, 1.1 mmol, 3 equiv.) were combined in chloroform (2 mL). The mixture was stirred at 50° C. for 1 h, during which time the white suspension became clear. Chloroform (200 mL) was added and the organic layer was gently washed twice with 1 M HCl (100 mL). After drying over MgSO4, filtration and removal of the solvent in vacuo, the compound was purified by column chromatography (flash SiO2) using an elution gradient of 5% to 40% MeOH in chloroform, which afforded pure DSPE-azidoacetate (244 mg, 0.29 mmol, 84%) as a white solid. 1H-NMR (400 MHz, CDCl3 / CD3OD 9:1): δ = 5.23 (dt, J = 9.0, 4.6 Hz, 1H), 4.35 (dd, J = 12.0, 3.7 Hz, 1H), 4.22-3.99 (m, 5H), 3.95 (s, 2H), 3.53 (t, J = 5.1 Hz, 2H), 2.33 (q, J = 7.6 Hz, 4H), 1.61 (td, J = 7.4, 4.2 Hz, 4H), 1.48-1.16 (m, 56H), 0.88 (t, J = 6.7 Hz, 6H). 13 C-NMR (101 MHz, CDCl3): δ = 173.7, 173.4, 168.3, 69.7, 69.6, 66.1, 66.0, 65.2, 62.1, 52.5, 40.01, 39.95, 34.3, 34.2, 34.1, 32.1, 29.9, 29.80, 29.7, 29.62, 29.59, 29.50, 29.47, 29.46, 29.4, 29.30, 29.26, 25.00, 24.97, 24.9, 22.8, 14.2. 31 P-NMR (162 MHz, CDCl3): δ = -0.48. MALDI-TOF MS: m / z C 43 H 83 Calculated value for N4O9P: 830.59; Found value [M+Na] + 853.62, [M-H+2Na] + 875.58.

[0286] MDP-Propargyl

[0287] [ka] A 50 mL round bottom flask was charged with MDP (0.113 g, 0.23 mmol, 1.00 equiv.) This material was dissolved in dry DMF (approximately 1.5 mL, 0.15 M) and the flask was purged with argon.

[0288] EDC.HCl (0.066 g, 0.34 mmol, 1.50 equiv.) and N,N-diisopropylethylamine (0.050 g, 0.068 mL, 0.39 mmol, 1.70 equiv.) and 4-(N,N-dimethylamino)pyridine (0.0028 g, 0.023 mmol, 0.10 equiv.) were added and the resulting clear solution was stirred at room temperature for 5 min. Then, prop-2-yn-1-amine (0.018 g, 0.021 mL, 0.32 mmol, 1.40 equiv.) was added via syringe. Stirring was continued at room temperature. After 21 h reaction time, LC-MS (water / MeOH) confirmed complete conversion of the MDP starting material. The reaction mixture was concentrated in vacuo to give the crude product as a yellow glass. This material was purified twice by automated column chromatography (reverse phase (C18); product: C18-silica 1:100; detection: 200-400 nm) eluting with water / MeOH 90 / 10-82 / 18. Pure fractions were lyophilized to give the pure product as a white solid (0.050 g, 41%). 1 H NMR (400 MHz, MeOD) δ 5.16 (d, J = 3.4 Hz, 1H), 4.42 - 4.23 (m, 3H), 3.95 (t, J = 2.3 Hz, 2H), 3.94 - 3.57 (m, 5H), 3.52 - 3.39 (m Hz, 1H), 2.58 (t, J = 2.6 Hz, 1H), 2.33 - 2.26 (m, 2H), 2.25 - 2.13 (m, 1H), 2.00 - 1.85 (m, 4H), 1.45 - 1.33 ppm (m, 6H). 13C NMR (100 MHz, MeOD) δ 175.26, 174.83, 173.89, 173.06, 172.08, 91.01, 79.15, 78.92, 76.68, 71.86, 70.83, 70.21, 63.35, 61.22, 54.13, 52.66, 49.48, 31.54, 28.11, 27.10, 23.85, 21.46, 18.31, 16.21 ppm. HPLC-MS (water / MeCN): t (product) = 0.76 and 1.02 min. Found: m / z = 512.08 [M-H2O+H] + ; 552.33 [M+Na] + (positive mode); 325.17 [M-Muramil]- (negative mode).

[0289] MDP(Bn)

[0290] [ka] MDP(Bn) was synthesized using standard SPPS techniques in a 100 mL glass reaction vessel equipped with a glass frit filter bottom. Adequate stirring of the reaction mixture was ensured by applying a constant argon flow through the glass frit filter, while excess reagents and washing solutions were removed by vacuum filtration. Crude MDP(Bn) was purified twice by automated column chromatography (reverse phase (C18); product: C18-silica 1:200; detection: 200-400 nm) eluted with water / MeCN+0.1% formic acid 90 / 10-82 / 18. Pure fractions were lyophilized to give the pure product as a fluffy white material (0.309 g, 67%). 1H NMR (400 MHz, DMF-d7) δ 8.18 (d, J = 8.6 Hz, 1H), 8.15 (d, J = 8.6 Hz, 1H), 7.75 (d, J = 6.5 Hz, 1H), 7.53 - 7.28 (m, 6H), 7.11 - 7.03 (m, 1H), 4.86 (d, J = 3.5 Hz, 1H), 4.76 (d, J = 12.3 Hz, 1H), 4.72 - 4.56 (m, 1H), 4.51 (d, J = 12.3 Hz, 1H), 4.47 - 4.32 (m, 3H), 4.01 (ddd, J = 10.7, 8.4, 3.5 Hz, 1H), 3.83 (dd, J = 11.6, 2.2 Hz, 1H), 3.78 - 3.61 (m, 3H), 3.60 - 3.40 (m, 1H), 2.39 (t, J = 7.8 Hz, 2H), 2.24 - 2.12 (m, 1H), 1.97 - 1.83 (m, 4H), 1.40 (d, J = 7.0 Hz, 3H), 1.34 ppm (d, J = 6.7 Hz, 3H). 13 C NMR (100 MHz, DMF-d7) δ 174.30, 173.74, 173.60, 172.82, 170.19, 138.47, 128.58, 127.93, 127.79, 97.05, 80.16, 77.36, 73.94, 70.66, 68.61, 61.76, 53.64, 52.59, 49.41, 35.63, 30.57, 30.47, 27.78, 22.65, 19.05, 18.04 ppm. HPLC-MS (water / MeCN): t(product) = 3.11 min. Measured value: m / z = 583.08 [M+H] + .

[0291] MDP(Bn)-プロパルギル

[0292]

change

[0293] MTP-b on resin

[0294]

change

[0295] MTP-b-N3

[0296] [ka] A 20 mL PE syringe equipped with a PE frit was loaded with MTP-b on resin (429 mg, approximately 0.15 mmol of MTP-b) and the resin was swollen in DMF (12 mL) for 30 min. The resin was treated twice for 15 min with a 2% solution of hydrazine hydrate in DMF (12 mL). After filtration, the resin was washed with DMF (4 × 12 mL) for 1 min. A solution of CuSO4·5H2O (0.6 mg, 2.4 μmol, 1.5 mol%), imidazole-1-sulfonyl azide HCl salt (170 mg, 0.77 mmol, 5 equiv.), and N,N-diisopropylethylamine (0.34 mL, 1.9 mmol, 12 equiv.) in DMF (12 mL) was added to the resin and the beads were stirred at room temperature for 24 h (a slight overpressure was released occasionally). After filtration, the resin was washed with DMF (5×12 mL) for 1 min and dichloromethane (4×10 mL) for 1 min. The resin was then subjected to cleavage in TFA / TIPS / HO 95:2.5:2.5 (4 mL) for 2 h. After filtration, the resin was washed with TFA (4 mL) for 5 min. The combined TFA filtrates were concentrated in vacuum (the temperature was kept as low as possible to avoid the formation of TFA esters). The impure compound was obtained by automated column chromatography (reverse phase (C18); detection: λ=200 nm) using an elution gradient of 5%-60% MeCN in HO (both containing 0.1% TFA). This was further purified by RP-HPLC using an elution gradient of 26%-35% MeCN in HO (both containing 0.1% TFA) to give the pure product (37.5 mg, 51 μmol, 34%) as a white fluffy solid after lyophilization. 1H-NMR (400 MHz, DMF-d7 / D2O 9:1): δ = 8.45 (t, J = 9.1 Hz, 2H), 8.26 (d, J = 7.9 Hz, 1H), 7.99 (d, J = 6.5 Hz, 1H), 7.81 (d, J = 2.8 Hz, 2H), 7.64-7.45 (m, 5H), 7.31 (d, J = 17.2 Hz, 2H), 5.82 (d, J = 6.3 Hz, 1H), 5.03 (d, J = 3.5 Hz, 1H), 4.93 (d, J = 12.4 Hz, 1H), 4.68 (d, J = 12.4 Hz, 1H), 4.63-4.46 (m, 4H), 4.19 (dd, J = 10.7, 3.6 Hz, 1H), 4.00 (dd, J = 6.8, 6.3 Hz, 1H), 3.92-3.79 (m, 3H), 3.66 (t, J = 9.0 Hz, 1H), 3.52 (t, J = 6.8 Hz, 2H), 2.53 (t, J = 7.5 Hz, 2H), 2.37 (dtd, J = 16.5, 7.9, 4.3 Hz, 1H), 2.10 (s, 3H), 2.09-1.93 (m, 2H), 1.88-1.55 (m, 5H), 1.58 (d, J = 7.1 Hz, 3H), 1.52 (d, J = 6.7 Hz, 3H). 13 C-NMR (100 MHz, DMF-d7 / D2O 9:1): δ = 174.9, 174.24, 174.17, 174.00, 173.93, 173.14, 173.06, 172.9, 172.8, 170.90, 170.8, 138.2, 128.6, 127.9, 127.8, 96.8, 80.0, 77.3, 73.7, 70.3, 68.6, 61.5, 53.6, 53.5, 53.2, 53.13, 53.10, 52.7, 52.6, 51.2, 49.4, 49.3, 32.1, 31.7, 28.5, 28.2, 23.2, 22.53, 22.48, 18.9, 17.80, 17.76. ESI-MS: m / z C 32 H 49 N9O 11Calculated value 735.36; Found value [M+H] + 736.25, [M+Na] + 758.42.

[0297] DSG 4-Nitrophenyl Carbonate

[0298] [ka] A 25 mL round-bottom flask was charged with a solution of commercially available [(2S)-3-hydroxy-2-octadecanoyloxypropyl]octadecanoate (0.601 g, 0.96 mmol, 1.00 equiv.) in chloroform (6.5 mL, ca. 0.15 M). Pyridine (0.122 g, 0.125 mL, 1.54 mmol, 1.60 equiv.) was added and the resulting clear solution was cooled in ice-water. Solid 4-nitrophenyl chloroformate (0.252 g, 1.25 mmol, 1.30 equiv.) was then added in portions. The pale yellow reaction mixture was stirred at room temperature overnight. Complete conversion of the alcohol was confirmed by precipitation (HCl, 100 ng / l ... 1 The reaction mixture was confirmed by H NMR (CDCl3). The reaction mixture was then precipitated in MeOH (100 mL) and collected by filtration through a glass filter. The material was washed with MeOH (total 30 mL) and Et2O (total 10 mL) and dried in a vacuum oven at 30°C. The product (0.713 g, 94%) was obtained as a white solid. 1 H NMR (400 MHz, CDCl3) δ 8.29 (d, J = 9.1 Hz, 2H), 7.39 (d, J = 9.1 Hz, 2H), 5.38 (p, J = 5.2 Hz, 1H), 4.50 (dd, J = 11.7, 3.9 Hz, 1H), 4.41 - 4.32 (m, 2H), 4.22 (dd, J = 12.0, 5.6 Hz, 1H), 2.35 (dt, J = 9.8, 7.5 Hz, 4H), 1.68 - 1.58 (m, 4H), 1.37 - 1.17 (m, 56H), 0.88 ppm (t, J = 6.7 Hz, 6H). 13C NMR (101 MHz, CDCl3) δ 173.24, 172.92, 155.35, 152.29, 145.56, 125.36, 121.76, 68.33, 66.97, 61.63, 34.16, 34.03, 31.93, 29.71, 29.68, 29.64, 29.49, 29.37, 29.29, 29.13, 29.07, 24.87, 22.70,14.12 ppm.

[0299] 2,3,5,6-Tetrafluorophenyl stearate

[0300] [ka] Triethylamine (2.8 mL, 5 equiv.) was added slowly to a solution of stearoyl chloride (1.26 g, 4.2 mmol) and tetrafluorophenol (0.72 g, 1.04 equiv.) in DCM (10 mL), resulting in immediate formation of a white precipitate. The heterogeneous reaction mixture was stirred for a further 2 h, diluted with 25 mL of DCM, extracted with water (50 mL), 0.1 M HCl (2×50 mL), dried over MgSO4 and evaporated to dryness. The resulting solid was redissolved in 50 mL of diethyl ether and extracted again with 1 M NaHCO3 (50 mL), 0.1 M HCl (50 mL), water (50 mL) and brine (2×50 mL). The organic phase was dried over MgSO4 (with the addition of a small amount of activated charcoal) and evaporated to dryness. The crude material was redissolved in chloroform, flashed onto a silica plug and evaporated to dryness again to give 1.2 g (67%) of the desired compound as a white solid. 1 H NMR (400 MHz, CDCl3) δ 6.98 (tt, J = 9.9, 7.0 Hz, 1H), 2.66 (t, J = 7.4 Hz, 2H), 1.78 (p, J = 7.4 Hz, 2H), 1.26 (s, 28H), 0.88 (t, J = 6.7 Hz, 3H) ppm. 19F NMR (376 MHz, CDCl3) δ -139.21 (ddd, J = 24.0, 11.8, 7.8 Hz), -153.03 - -153.20 (m). 13 C NMR (101 MHz, CDCl3) δ 169.56, 147.23 (m), 144.74 (m), 141.94, 141.86 (m), 139.41 (m), 129.78 (m), 103.01 (t), 33.42, 31.92, 29.69, 29.65, 29.61, 29.54, 29.39, 29.36, 29.14, 28.85, 24.78, 22.68, 14.09 ppm.

[0301] synthetic アプローチ First approach. The compounds of the invention can be prepared by using the starting reactants MDP or MDP(Bn) (see above). These molecules have a functional carboxylic acid group derived from the glutamic acid (Glu) building block. The COOH group allows conjugation to amine-functional reactants containing lipophilic groups. Such lipophilic groups can be, for example, C18 moieties, such as those derived from stearic acid, oleic acid, stearyl alcohol, oleyl alcohol, stearylamine or oleylamine; or sterol moieties, such as those derived from cholesterol. Saturated linear lipophilic moieties are preferred, as are those derived from cholesterol. Particularly useful building blocks are PE phospholipids, such as DSPE ([1069-79-0]) or DOPE ([4004-05-1]); these molecules are already amine-functionalized. Mixed acyl PE-phospholipids can also be useful (e.g., 16:0 to 18:1 PE, or 18:0 to 18:1 PE, or 18:0 to 16:0 PE). Another useful building block is cholesterol. Yet another useful building block is diglyceride, such as 1,2-dioctadecanoyl-sn-glycerol (18:0 DG [51063-97-9]) or 1-2-dioleoyl-sn-glycerol (18:1 DG [24529-88-2]). Mixed acyl diglycerides can also be used (e.g., 16:0 to 18:1 DG, or 18:0 to 18:1 DG, or 18:0 to 16:0 DG). PE-phospholipids and diglycerides have two lipophilic chains, and such building blocks are preferably used in this approach. This first approach is illustrated in Examples 10-19.

[0302] Second approach. A particularly suitable alternative modular approach uses copper-catalyzed azide-alkyne cycloaddition ("click reaction") to connect MDP (or MTP) reactants to lipophilic reactants. Here, MDP, MDP(Bn), MTP or MTP(Bn) building blocks with azide (-N3) or alkyne (-C≡CH) functional groups are used. Non-limiting examples of such molecules are MDP-propargyl, MDP(Bn)-propargyl or MTP-b-N3 (see above). In a copper-catalyzed click reaction, these molecules can be coupled with alkyne- or azide-functional molecules that contain lipophilic groups. Using the click reaction, modular, easily prepared and stable azide- or alkyne-functional intermediates are targeted. This allows for easy isolation and storage of the intermediates. Furthermore, the copper-catalyzed click cycloaddition reaction can be and is best carried out in an aqueous environment (e.g., THF / water or tBuOH / water, etc.) or in an aqueous biphasic liquid / liquid solvent combination (e.g., dichloromethane / water, etc.). In these reaction media, both the hydrophilic MDP (or MTP) reactant (with or without a Bn group) and the lipophilic reactant are conveniently dissolved, greatly improving the ease of conjugation and the reaction yield. In this click approach, the lipophilic group contained in the azide or alkyne reactant can be a C14, C16, or C18 moiety, such as stearic acid, palmitic acid, myristic acid, oleic acid, palmitoleic acid, myristoleic acid, stearyl alcohol or amine, palmityl alcohol or amine, myristyl alcohol or amine, oleyl alcohol or amine, palmitoleic alcohol or amine, myristoleic alcohol or amine; or a sterol moiety, such as one derived from cholesterol. Saturated linear lipophilic moieties are preferred, as are moieties derived from cholesterol. Particularly useful building blocks are PE-phospholipids containing C14, C16 and / or C18 moieties, mixed acyl PE-phospholipids, diglycerides (DG) or mixed acyl diglycerides, and cholesterol.Lipophilic azide or alkyne reactants containing two lipophilic chains or containing a cholesteryl group are preferred. This second approach is illustrated in Examples 1-9.

[0303] It should be noted that both approaches allow the introduction of an additional amino acid unit linked to the glutamic acid unit of MDP or MDP(Bn). The preferred amino acid units are derived from L-lysine or L-alanine. When the additional amino acid unit is attached, the MDP (muramyl dipeptide) moiety is converted to an MTP (muramyl tripeptide) moiety, either with or without a Bn group. Examples are shown in Examples 9-10, 13-15, and 17-18.

[0304] [Example 1] Synthesis of MDP-C18 [click] (1)

[0305] [ka] Molecular weight: 825 Daltons. CLogP=4.15. This synthesis demonstrates the general conditions for Cu click-type reactions.

[0306] A 5 mL vial was charged with MDP-propargyl (0.011 g, 0.02 mmol, 1.00 equiv). To this was added L-ascorbic acid (0.4 M in water, 104 μL, 41.5 μmol ascorbic acid, 2.00 equiv). To the resulting slightly opaque solution was added a solution of 1-azidooctadecane (0.012 g, 0.04 mmol, 2.00 equiv) in DCM (0.8 mL), followed by aqueous copper(II) sulfate pentahydrate (0.2 M, 104 μL, 20.8 μmol Cu, 1.00 equiv). The biphasic reaction mixture was then stirred at 1400 rpm at room temperature to give a pale yellow / green emulsion. After 16 h, the reaction mixture was concentrated in a stream of N2 to give the crude product as a light brown sludge. This material was dissolved in chloroform / MeOH 4:1 and impregnated onto Celite (90 mg, approximately 1:5 loading ratio) and purified by automated column chromatography (product:silica 1:500; detection: 200-400 nm) eluting with chloroform / MeOH / water 90 / 9 / 1 to 70 / 27 / 3 to give the product (0.005 g, 31%) as a white solid. 1 H NMR (400 MHz, MeOD) δ 7.64 - 7.61 (m, 1H), 5.33 (d, J = 3.4 Hz, 1H), 4.58 - 4.21 (m, 7H), 3.85 - 3.46 (m, 6H), 2.35 - 2.26 (m, 2H), 2.23 - 2.08 (m, 1H), 2.06 - 1.84 (m, 6H), 1.42 - 1.36 (m, 6H), 1.35 - 1.22 (m, 30H), 0.88 ppm (t, J = 6.8 Hz, 3H). 13C NMR (100 MHz, MeOD) δ 175.75, 174.66, 174.04, 173.58, 171.91, 144.63, 122.53, 91.00, 76.01, 71.84, 71.18, 67.16, 61.98, 54.15, 52.98, 50.67, 34.78, 32.20, 32.03, 30.34, 29.80, 29.76, 29.72, 29.65, 29.52, 29.46, 29.12, 27.49, 26.60, 22.83, 22.79, 22.69, 19.34, 16.95, 16.57, 14.15 ppm. HPLC-MS (water / MeCN): t (product) = 5.64 min. Observed value: m / z = 825.33 [M+H]+.

[0307] [Example 2] Synthesis of MDP-DSPE [click] (2)

[0308] [ka] Molecular weight: 1361 Daltons. CLogP=11.56 (uncharged) and 5.78 (negatively charged).

[0309] MDP-propargyl (24.4 mg, 46 μmol) was dissolved in 0.4 M ascorbic acid (0.24 mL, 2 equiv.) and a solution of DSPE-azidoacetate (38.5 mg, 46 μmol, 1 equiv.) in dichloromethane (0.5 mL) was added. Under vigorous stirring, 0.2 M CuSO4·5H2O (0.24 mL, 1 equiv.) was added and the biphasic system was stirred vigorously for 19 h at room temperature. The solvent was removed in vacuo and the greenish solid was subjected to column chromatography (flash SiO2) using an elution gradient of 20% to 50% MeOH in chloroform, ending with (45% MeOH + 5% H2O) in chloroform (a significant amount of compound is eluted only after the addition of H2O). This gave an impure product, which was purified by automated column chromatography (reverse phase (C 18 );Product:C 18-silica 1:200; detection: λ = 210 nm) using an elution gradient of 25% to 70% THF in HO, which gave the pure product (19.5 mg, 14 μmol, 31%) as a white fluffy solid after lyophilization. 1 H NMR (400 MHz, CDCl3+MeOD) δ 7.93 (s, 1H), 5.29 - 5.22 (m, 2H), 5.16 (s, 2H), 4.51 - 4.41 (m, 4H), 4.38 - 4.25 (m, 2H), 4.20 (dd, J = 12.1, 6.8 Hz, 1H), 4.05 - 3.89 (m, 4H), 3.88 - 3.77 (m, 2H), 3.77 - 3.63 (m, 6H), 3.55 - 3.42 (m, 3H), 2.34 (q, J = 7.3 Hz, 6H), 2.22 (dddd, J = 18.4, 13.5, 8.4, 5.7 Hz, 1H), 1.98 (d, J = 5.7 Hz, 4H), 1.93 (s, 0H), 1.62 (q, J = 6.4 Hz, 5H), 1.46 - 1.36 (m, 6H), 1.28 (s, 61H), 0.89 (t, J = 6.8 Hz, 6H). The peak between 4.9 and 4.6 ppm is not visible due to overlap with the HO peak. MALDI-TOF MS: m / z C 65 H 118 N9O 19 Calculated P 1359.83; Found [M+Na] + 1382.83, [M-H+2Na] + 1404.84. HPLC-MS (H2O / THF, gradient: 65-95% THF): t (product) = 2.33 min; m / z = 1360.80 [M+H] + (SIM mode).

[0310] [Example 3] Synthesis of MDP-chol [click] (3)

[0311] [ka] Molecular weight: 999 Daltons. CLogP=5.04.

[0312] MDP-propargyl (25 mg, 47 μmol) was dissolved in 0.4 M ascorbic acid (0.24 mL, 2 equiv.) and cholesterol azide acetate (26.6 mg, 57 μmol, 1.2 equiv.) in dichloromethane (0.5 mL) was added. Under vigorous stirring, 0.2 M CuSO4·5H2O (0.24 mL, 1 equiv.) was added and the biphasic system was stirred vigorously for 17 h at room temperature. H2O / brine 1:1 (50 mL) was added and the bluish aqueous layer was extracted with chloroform / MeOH 2:1 (5 × 20 mL). The combined organic layers were dried using Na2SO4, filtered and the solvent was removed in vacuo. The resulting colorless solid was purified by repeated column chromatography (flash SiO2) using an elution gradient of 6%–20% MeOH in chloroform. This gave the pure product (24.4 mg, 24 μmol, 52%) as a white fluffy solid after lyophilization from THF / H2O. 1 H-NMR (400 MHz, THF-d8 / D2O 95:5): δ = 7.81 (s, 1H), 5.28 (d, J = 4.9 Hz, 1H), 5.16 (s, 2H), 5.10 (d, J = 3.4 Hz, 1H), 4.59-4.46 (m, 1H), 4.46-4.16 (m, 5H), 3.75-3.17 (m, 6H), 2.26 (d, J = 8.2 Hz, 2H), 2.20 (t, J = 7.5 Hz, 2H), 2.12-1.99 (m, 1H), 1.98-1.69 (m, 8H), 1.58-1.33 (m, 5H), 1.32-1.23 (m, 7H), 1.18 (s, 5H), 1.13-0.97 (m, 4H), 0.94 (s, 3H), 0.84 (d, J = 6.5 Hz, 3H), 0.77 (dd, J = 6.6, 1.4 Hz, 8H), 0.61 (s, 3H). 13C-NMR (100 MHz, THF-d8 / D2O 95:5): δ = 175.2, 174.9, 173.7, 173.4, 171.9, 166.6, 165.1, 139.6, 122.4, 91.0, 78.7, 76.7, 75.4, 72.0, 70.2, 61.1, 56.8, 56.2, 54.0, 52.6, 50.5, 50.2, 49.5, 42.2, 39.8, 39.4, 37.7, 36.8, 36.4, 36.1, 35.8, 34.4, 31.9, 31.8, 29.6, 28.1, 27.9, 27.5, 27.4, 22.5, 22.2, 22.1, 21.9, 20.9, 18.8, 18.7, 18.2, 16.8, 13.5, 11.3. 51 H 82 N8O 12 Calculated value 998.60; Found value [M+Na] + 1021.58. HPLC-MS (H2O / THF, gradient: 65-95% THF): t (product) = 2.20 min; m / z = 999.60 [M+H] + (SIM mode).Note: THF-d8 / D2O 95:5 was found to be the best solvent combination for NMR characterization. Nevertheless, the spectra have overlaps and are very complex. Therefore, the integration is tentative.

[0313] [Example 4] Synthesis of MDP-DSPE2 [click] (4)

[0314] [ka] Molecular weight: 2192 daltons.

[0315] [Example 5] MDP-Chol2[click]

[0316] [ka] Molecular weight: 1669 Daltons. CLogP=15.31.

[0317] [Example 6] MDP-DSG[Click](6)

[0318] [ka] Molecular weight: 1238 daltons. CLogP=12.45.

[0319] [Example 7] MDP(Bn)-DSPE[click](7)

[0320] [ka] Molecular weight: 1451 Daltons. CLogP=13.85 (uncharged) and 8.06 (negatively charged).

[0321] Following the general conditions for Cu-click reaction, MDP(Bn)-propargyl (0.028 g, 0.045 mmol, 1.00 equiv.) and DSPE-azidoacetate (0.039 g, 0.047 mmol, 1.05 equiv.) were reacted overnight. During the reaction, some material precipitated, resulting in a white suspension / emulsion. The reaction mixture was then diluted with chloroform / MeOH 1:1. The resulting clear solution was impregnated onto Celite (approximately 200 mg, loading ratio 1:3). The impregnated crude product was first purified by automated column chromatography (reverse phase (C18); product: C18-silica 1:200; detection: ELSD and UV 200-400 nm) eluted with water / THF 60 / 40-20 / 80. The combined product fractions were lyophilized and then purified again by automated column chromatography (normal phase (silica); product: silica 1:300; detection: ELSD) eluting with chloroform / MeOH / water 90 / 9 / 1 to 75 / 22.5 / 2.5. The pure fractions were concentrated in vacuo, dissolved in water / THF 70 / 30 and lyophilized. In this way the pure product was obtained as a white fluffy solid (0.027 g, 41%). 11H NMR (400 MHz, CDCl3 + MeOD 1:1) δ 7.99 (d, J = 8.2 Hz, 1H), 7.90 (s, 1H), 7.43 - 7.23 (m, 5H), 5.24 (m, 1H), 5.13 (s, 2H), 4.93 (d, J = 3.5 Hz, 1H), 4.73 (d, J = 12.0 Hz, 1H), 4.51 (d, J = 12.0 Hz, 2H), 4.48 - 4.40 (m, 2H), 4.37 - 4.15 (m, 3H), 4.05 - 3.89 (m, 5H), 3.86 - 3.73 (m, 2H), 3.71 - 3.53 (m, 3H), 3.49 - 3.42 (m, 2H), 2.37 - 2.28 (m, 6H), 2.25 - 2.11 (m, 1H), 2.05 - 1.89 (m, 4H), 1.67 - 1.56 (m, 4H), 1.52 - 1.19 (m, 72H), 0.89 ppm (t, J = 6.8 Hz, 6H). 13C NMR (100 MHz, MeOD) δ 174.97, 174.73, 173.93, 173.66, 173.57, 173.46, 171.92, 166.34, 137.16, 128.28, 128.09, 127.84, 124.43, 96.38, 79.09, 76.52, 72.55, 70.40, 69.75, 69.28, 63.66, 63.44, 62.56, 61.14, 53.18, 52.60, 52.13, 49.45, 40.52, 34.62, 34.12, 33.96, 31.81, 31.73, 29.57, 29.53, 29.43, 29.41, 29.23, 29.21, 29.03, 29.00, 27.12, 24.82, 24.77, 22.52, 22.20, 18.70, 16.75, 13.69 ppm. MALDI-TOF: m / z C 72 H 124 N9O 19 P + 2Na + -H + Calculated value of: 1494.85 [M + 2Na - H] +; Found: 1494.94. HPLC-MS (water / THF, gradient: 55-95% THF): t (product) = 5.09 min; m / z = 1450.9 [M+H] + and 1472.9 [M+Na] + (SIM mode).

[0322] [Example 8] Synthesis of MDP(Bn)-chol [click] (8)

[0323] [ka] Molecular weight: 1089 daltons. CLogP=6.83.

[0324] Following the general conditions for Cu-click reaction, MDP(Bn)-propargyl (0.030 g, 0.048 mmol, 1.00 equiv.) and cholesterol azide acetate (0.025 g, 0.053 mmol, 1.10 equiv.) were reacted overnight to give a white emulsion. The reaction mixture was then concentrated in vacuum and impregnated onto Celite (150 mg). The impregnated crude product was purified by automated column chromatography (reverse phase (C18); product: C18-silica 1:200; detection: 200-400 nm) eluting with water / THF 70 / 30-15 / 85. The combined product fractions were lyophilized and then purified again by automated column chromatography (normal phase (silica); product: silica 1:350; detection: ELSD) eluting with chloroform / MeOH 96 / 4-86 / 14. The pure fractions were concentrated in vacuo to give the pure product as a white solid (0.028 g, 53%). 1<h2 style=";text-align:left;direction:ltr">H NMR (400 MHz, CDCl3+MeOD 1:1) δ 7.78 (s, 1H), 7.39 - 7.28 (m, 5H), 5.38 (d, J = 5.1 Hz, 1H), 5.16 (d, J = 1.6 Hz, 2H), 4.93 (d, J = 3.6 Hz, 1H), 4.70 (d, J = 11.8 Hz, 2H), 4.56 - 4.37 (m, 3H), 4.30 (dt, J = 8.9, 4.4 Hz, 1H), 4.27 - 4.17 (m, 2H), 4.02 (dd, J = 10.1, 3.6 Hz, 1H), 3.93 - 3.67(m, 24H), 3.67 - 3.53 (m, 3H), 3.40 (d, J = 3.2 Hz, 0H), 2.44 - 2.32 (m, 2H), 2.27 (td, J = 7.1, 3.7 Hz, 2H), 2.12 (dtd, J = 14.9, 7.5, 4.3 Hz, 1H), 2.07 - 1.74 (m, 8H), 1.72 - 1.42 (m, 5H), 1.38 (dd, J = 13.0, 7.0 Hz, 7H), 1.34 - 0.94 (m, 13H), 0.92 (d, J = 6.4 Hz, 3H), 0.87 (dd, J = 6.6, 1.8 Hz, 6H), 0.69 ppm (s, 3H).<h2 style=";text-align:left;direction:ltr"> 13C NMR (100 MHz, CDCl3+MeOD 1:1) δ 175.17, 174.60, 173.71, 173.62, 171.76, 166.21, 145.18, 139.13, 137.21, 128.65, 128.37, 128.25, 124.35, 123.47, 96.91, 79.14, 76.83, 76.49, 72.44, 69.79, 61.63, 56.82, 56.28, 53.17, 52.57, 51.14, 50.14, 49.78, 42.45, 39.84, 39.65, 38.01, 36.97, 36.68, 36.32, 35.93, 34.85, 32.02, 31.97, 29.81, 28.34, 28.14, 27.74, 27.58, 24.39, 23.95, 22.87, 22.80, 22.77, 22.61, 21.16, 19.34, 18.98, 18.80, 16.90, 11.94 ppm. MALDI-TOF: m / z 58 H 88 N8O 12 +Na + Calculated value: 1111.64 [M+Na] + ; Found: 1111.65. HPLC-MS (water / THF, gradient: 65-95% THF): t (product) = 2.79 min; m / z = 1089.70 [M+H] + (SIM mode).

[0325] [Example 9] Synthesis of MTP-b-C18[invclick] (9)

[0326] [ka] Molecular weight: 1058 Daltons. CLogP=5.68.

[0327] MTP-b-N3 (26 mg, 35 μmol) and prop-2-yn-1-yl stearate (11.3 mg, 35 μmol, 1 equiv.) were suspended in THF (0.36 mL) and 0.4 M ascorbic acid (0.18 mL, 2 equiv.) was added to give a clear solution. Prop-2-yn-1-yl stearate was prepared using known procedures. Under vigorous stirring, 0.2 M CuSO4·5H2O (0.18 mL, 1 equiv.) was added and the mixture was stirred vigorously at room temperature (gelling occurred initially, but gentle heating gave a yellow solution). After 1 h, HPLC-MS (THF / H2O) showed the absence of starting compound and the opaque solution was lyophilized. The crude product was adsorbed onto Celite from chloroform / MeOH 2:1 and subjected to column chromatography (flash SiO2) using an elution gradient of 10% to 25% MeOH in chloroform. Column chromatography was repeated using a similar gradient to give the pure product (31.5 mg, 30 μmol, 84%) as a white fluffy solid after lyophilization from THF / H2O. 1H-NMR (400 MHz, THF-d8 / D2O 4:1): δ = 7.92 (s, 1H), 7.31 (d, J = 7.5 Hz, 2H), 7.23 (t, J = 7.5 Hz, 2H), 7.15 (t, J = 7.3 Hz, 1H), 5.04 (s, 2H), 4.74 (d, J = 3.5 Hz, 1H), 4.63 (d, J = 12.2 Hz, 1H), 4.40 (d, J = 12.2 Hz, 1H), 4.31 - 4.15 (m, 6H), 3.96 (dd, J = 10.5, 3.6 Hz, 1H), 3.66 (d, J = 3.2 Hz, 2H), 3.59 - 3.45 (m, 3H), 2.22 (dt, J = 15.3, 7.7 Hz, 4H), 2.09 (tt, J = 12.9, 6.0 Hz, 1H), 1.87 - 1.70 (m, 6H), 1.64 - 1.54 (m, 1H), 1.47 (q, J = 7.3 Hz, 2H), 1.32 (d, J = 7.2 Hz, 3H), 1.28 (d, J = 6.7 Hz, 3H), 1.18 (s, 30H), 0.78 (t, J = 6.6 Hz, 3H). 13 C-NMR (100 MHz, THF-d8 / D2O 4:1): δ = 175.5, 174.9, 174.8, 173.73, 173.66, 173.2, 171.7, 142.3, 137.8, 128.1, 128.0, 127.4, 124.3, 96.6, 80.0, 77.3, 72.8, 69.1, 68.8, 60.9, 57.2, 53.2, 53.1, 52.2, 49.7, 49.5, 33.6, 31.8, 31.5, 30.9, 29.7, 29.54, 29.50, 29.4, 29.24, 29.20, 29.0, 27.7, 22.54, 22.49, 21.9, 18.6, 16.9, 13.5. MALDI-TOF MS: m / z C 53 H 87 N9O 13 Calculated value of 1057.64; Measured value [M+Na] + 1080.63, [M+K] +1096.65. HPLC-MS (H2O / THF, gradient: 65-95% THF): t (product) = 2.15 min; m / z = 1058.60 [M+H] + (SIM mode).

[0328] [Example 10] Synthesis of MTP-b-C18(10)

[0329] [ka] Molecular weight: 976 Daltons. CLogP=5.31

[0330] A 10 mL PE syringe equipped with a PE frit was loaded with MTP-b on resin (137 mg, approximately 0.0493 mmol of MTP-b, 1.00 equiv.). The resin was allowed to swell in DMF (5 mL) for 30 min. The resin was then treated twice for 15 min with a 2% solution of hydrazine hydrate in DMF (10 mL). The hydrazine solution was removed and the resin was washed with DMF (4×5 mL). A solution of 2,3,5,6-tetrafluorophenyl stearate (0.064 g, 0.15 mmol, 3.00 equiv.) and 4-methylmorpholine (0.030 g, 0.033 mL, 0.30 mmol, 6.00 equiv.) in DMF / DCM (1+1 mL; approximately 0.075 M) was then added. The beads were stirred overnight at room temperature. The supernatant was then removed and the resin was washed with DMF / DCM 50 / 50 (4 x 5 mL) and DCM (2 x 5 mL). The resin was then treated with TFA / TIPS / water 95 / 2.5 / 2.5 (200 μL) for 1 h. The filtrate was collected and the resin was washed with additional cleavage cocktail. The combined filtrates were concentrated in vacuo to give the crude product as a white solid. This material was impregnated onto Celite (200 mg, loading ratio 1:4) from THF / water solution (95 / 5). The impregnated crude product was purified by automated column chromatography (reverse phase (C18); product: C18-silica 1:250; detection: ELSD) eluting with water / THF 50 / 50 to 10 / 90. The combined product fractions were lyophilized and then purified again by automated column chromatography (normal phase (silica); product: silica 1:500; detection: ELSD) eluting with dichloromethane / MeOH 90 / 10 to 70 / 30. Pure fractions were concentrated in vacuo to give the product as a white solid (0.016 g, 33%). 1H NMR (400 MHz, CDCl3+TFA-d3) δ 7.40 - 7.24 (m, 5H), 4.94 - 4.89 (m, 1H), 4.71 - 4.65 (m, 1H), 4.57 - 4.17 (m, 6H), 4.03 - 3.74 (m, 4H), 3.38 (bs, 2H), 2.60 - 2.22 (m, 5H), 2.30 (s, 1H), 2.11 - 1.55 (s, 11H), 1.50 - 1.17 (m, 36H), 0.87 ppm (t, J = 6.6 Hz, 3H). HPLC-MS (Water / MeCN): t (product) = 5.64 min. Actual value: m / z = 976.33 [M+H] + (Sun mode); 1020.25 [M+HCOO] - (Shadow mode).

[0331] [Example 11] Synthesis of MDP-C18 (11)

[0332] [ka] Molecular weight: 744 Daltons. CLogP=4.79.

[0333] MDP (10 mg, 20 μmol), octadecylamine (5.2 mg, 0.95 equiv), NHS (2.4 mg, 1 equiv) and EDC-HCl (7.9 mg, 2 equiv) were stirred in DMF (0.7 mL) at 50° C. for 3 h. The reaction mixture was then allowed to cool to room temperature and stirred for an additional 16 h. The resulting dispersion was heated to 40° C. to redissolve any precipitated solids and then precipitated with 5 mL of ether. The collected precipitate was washed two more times with ether, dried, then suspended in demineralized water, collected by centrifugation, resuspended in demineralized water and collected again by centrifugation. The resulting solid was freeze-dried to remove all water, yielding 13.8 mg (96%) of the desired compound as a white powder. 1H NMR (400 MHz, DMF-d7) δ 8.31 (d, J = 7.7 Hz, para-isomer), 8.23 ​​(d, J = 8.0 Hz, primary isomer), 8.12 (d, J = 7.9 Hz, para-isomer), 8.09 (d, J = 7.6 Hz, primary isomer), 7.96 (d, J = 6.6 Hz, para-isomer), 7.90 (d, J = 6.5 Hz, primary isomer), 7.77 (m, 1H), 7.47 (m, 1H), 7.10 (m, para-isomer), 7.01 (m, primary isomer), 6.86 (d, J = 6.0 Hz, paraisomer), 6.74 (dd, J = 4.1, 1.2 Hz, primary isomer), 5.44 - 5.29 (m, 1H), 5.16 (t, J = 3.7 Hz, primary isomer), 4.79 (t, J = 6.1 Hz, paraisomer), 4.60 (dd, J = 8.2, 6.0 Hz, paraisomer), 4.57-4.22 (m, 4H), 3.93-3.57 (m, 5H), 3.45 (m, 1H), 3.13 (m, 2H), 2.42-2.07 (m, 3H), 2.00-1.78 (m, 4H), 1.56 - 1.07 (m, 38H), 0.88 (m, 3H) ppm. 13 C NMR (101 MHz, DMF-d7) δ 174.25, 174.20, 173.93, 173.89, 172.75, 172.56, 172.07, 172.02, 171.76, 170.11, 96.92, 91.66, 82.46, 79.55, 77.39, 77.00, 72.97, 71.30, 70.95, 62.05, 57.49, 54.49, 53.16, 53.03, 49.54, 39.26, 32.55, 32.05, 29.83, 29.49, 28.62, 28.41, 27.18, 22.86, 22.76, 22.73, 19.19, 17.80, 17.62, 13.93 ppm. ESI-MS: m / z 743.50 (calculated value), measured value 744.42 (M+H + ), 788.33 (M-FA - ).

[0334] [Example 12] Synthesis of MDP-DSPE (12)

[0335] [ka] Molecular weight: 1223 Daltons. CLogP=12.96 (uncharged) and 7.18 (negatively charged).

[0336] MDP (14 mg, 29 μmol), NHS (5.6 mg, 1.7 equiv.), and DIC (7.3 mg, 2 equiv.) were stirred in 0.9 mL of DMF for 2 h to activate MDP. The resulting mixture was added to a dispersion of DSPE (17 mg, 0.8 equiv.) in 2.7 mL of tert-butanol containing TEA (9 mg, 3.1 equiv.) at 50 °C and stirred at that temperature for 3.5 h. The resulting mixture was evaporated to dryness and the resulting material was repeatedly purified by column chromatography (SiO2, CHCl3 / MeOH / H2O, 70 / 30 / 5, 5:4:1 and gradient 95 / 5 / 0 to 60 / 40 / 0) to give 6 mg (21%) of the desired compound as a white fluffy material after lyophilization from water / THF. 1 H-NMR (400 MHz, CDCl3 / CD3OD 5:) δ 5.29 (d, major isomer), 5.24 (m, 1H), 4.54 (d, minor isomer), 4.47 (m, obscured by HDO), 4.41 (dd, obscured by HDO), 4.31 (m, obscured by HDO), 4.19 (dd, 1H), 4.00-3.90 (m, mixture of isomers), 3.87-3.75 (m, mixture of isomers), 3.72 (m, 1H), 3.63 (m, 1H), 3.53-3.30 (m, obscured by CD3OD), 2.37-2.26 (m, 5H), 2.25-2.10 (m, 2H), 2.10-1.92 (m, 4H), 1.61 (m, 4H), 1.50-1.15 (br.m, 62H), 0.88 (t, 6H) ppm. 31 P-NMR (162 MHz, CDCl3 / CD3OD 5:1) δ 0.14 (br.m) ppm.13 C-NMR (101 MHz, CDCl3 / CD3OD 5:1) δ 174.63, 174.00, 173.27, 173.08, 172.92, 172.89, 171.34, 90.19, 75.19, 71.09, 70.18, 69.73, 69.65, 63.24, 62.76, 61.86, 60.84, 53.13, 52.04, 48.77, 48.19, 39.71, 33.48, 33.32, 31.29, 31.14, 28.91, 28.87, 28.76, 28.74, 28.57, 28.54, 28.37, 28.34, 26.44, 24.14, 24.10, 21.88, 21.78, 18.35, 15.98, 13.14 ppm. MALDI-MS: m / z 1221.77 (calculated), found 1220.82 (MH - ), negative mode. HPLC-ELSD (C18, 65-95% THF / H2O): single peak + shoulder for alpha and beta isomers.

[0337] [Example 13] Synthesis of MTP-b-DSG (13)

[0338] [ka] Molecular weight: 1361 daltons. CLogP=15.05.

[0339] A 20 mL PE syringe equipped with a PE frit was loaded with MTP-b on resin (0.291 g, approximately 0.11 mmol of MTP-b, 1.00 equiv.). The resin was allowed to swell in DMF (5 mL) for 30 min. The resin was then treated twice for 15 min with a 2% solution of hydrazine hydrate in DMF (5 mL). The hydrazine solution was removed and the resin was washed with DMF (4×5 mL). A solution of DSG 4-nitrophenyl carbonate (0.249 g, 0.32 mmol, 3.00 equiv.) and N,N-diisopropylethylamine (0.081 g, 0.110 mL, 0.63 mmol, 6.00 equiv.) in chloroform (4 mL) was then added. The beads were stirred overnight at room temperature. The bright yellow supernatant was then removed and the resin was washed with chloroform (3 × 5 mL), MeOH (3 × 5 mL), and again with chloroform (2 × 5 mL). The resin was treated with TFA / TIPS / water 95 / 2.5 / 2.5 (5 mL) for 45 min. The supernatant was poured into ice-cold diethyl ether (100 mL) with stirring, which slowly formed white aggregates. This cleavage and precipitation procedure was repeated twice. The solids were collected by filtration through a disposable PE filter, giving the crude product as a white solid. This material was impregnated onto Celite (250 mg, loading ratio 1:2.5) from a chloroform / MeOH (1:2) solution. The impregnated crude product was purified by automated column chromatography (reverse phase (C18); product: C18-silica 1:150; detection: 200–400 nm) eluted with water / THF 60 / 40–0 / 100. The combined product fractions were lyophilized and then purified again by automated column chromatography (normal phase (silica); product:silica 1:250; detection: 200-400 nm) eluting with chloroform / MeOH 95 / 5-60 / 40. Pure fractions were concentrated in vacuo to give the product as a white solid (0.030 g, 21%). 1H NMR (400 MHz, CDCl3+MeOD) δ 7.39 - 7.27 (m, 5H), 5.25 (p, J = 5.3 Hz, 1H), 4.89 (d, J = 3.5 Hz, 1H), 4.73 (d, J = 11.9 Hz, 1H), 4.50 (d, J = 11.9 Hz, 1H), 4.42 - 4.31 (m, 2H), 4.31 - 4.19 (m, 4H), 4.15 (dd, J = 11.9, 6.2 Hz, 2H), 4.10 - 3.97 (m, 1H), 3.87 - 3.75 (m, 2H), 3.72 - 3.53 (m, 3H), 3.12 (t, J = 6.9 Hz, 2H), 2.37 - 2.27 (m, 6H), 2.26 - 2.13 (m, 1H), 1.93 (s, 3H), 1.92 - 1.74 (m, 2H), 1.71 - 1.56 (m, 6H), 1.56 - 1.47 (m, 2H), 1.43 (d, 7.0 Hz, 3H), 1.40 (d, 7.0 Hz, 3H), 1.36 - 1.19 (s, 56H), 0.89 ppm (t, J = 6.7 Hz, 6H). 13C NMR (100 MHz, CDCl3+MeOD) δ 175.80, 174.79, 174.30, 173.86, 173.76, 173.44, 173.25, 171.70, 156.58, 137.07, 128.36, 128.14, 127.95, 96.52, 79.36, 76.83, 72.38, 69.48, 69.43, 69.37, 62.50, 62.29, 61.30, 53.52, 53.24, 51.71, 49.46, 40.33, 34.13, 33.97, 31.82, 31.29, 31.08, 29.58, 29.54, 29.52, 29.41, 29.39, 29.25, 29.20, 29.18, 29.09, 29.01, 28.98, 28.16, 24.80, 24.77, 22.88, 22.55, 22.35, 18.61, 16.89, 13.77 ppm. HPLC-MS (water / THF, gradient: 55-95% THF): t (product) = 6.20 min. Observed: m / z = 1360.9 [M+H] + and 1382.9 [M+Na] + (SIM mode).

[0340] [Example 14] Synthesis of MTP(Bn)-a-DPPE (14)

[0341] [ka] Molecular weight: 1328 Daltons. CLogP = 12.88 (uncharged) and 7.09 (negatively charged)

[0342] Building block (CBz)-Ala-DPPE N-CBz-protected L-alanine (390 mg, 1.7 mmol) and N-hydroxysuccinimide (222 mg, 1.89 mmol, 1.1 equiv.) were dissolved in chloroform (6 mL) to obtain a nearly clear solution. N,N'-diisopropylcarbodiimide (DIC; 0.32 mL, 2.0 mmol, 1.2 equiv.) was added and the mixture was stirred at room temperature for 40 min (after 1 min, the solution became cloudy, after 25 min, 1H-NMR showed complete conversion). This solution was then added to a 60 °C solution containing 1,2-dipalmitoyl-sn-glycero-3-phosphoethanolamine (DPPE; 1.06 g, 1.5 mmol, 0.9 equiv.) and triethylamine (600 μL, 0.71 mmol, 2.5 equiv.) in chloroform (12 mL; DPPE was dissolved under reflux and triethylamine was added at low temperature). The resulting clear solution was stirred at 60° C. for 1 h (the solution remained clear and 1 H-NMR showed complete conversion). Chloroform (360 mL) was added and the organic layer was gently washed with 0.1 M HCl (100 mL). The organic layer was dried using Na2SO4, filtered and the solvent removed in vacuo. Column chromatography (flash SiO2) using an elution gradient of 2% to 30% methanol in chloroform afforded the title compound partially contaminated with triethylamine. Impure fractions were dissolved in chloroform and the organic layer was gently washed with 0.1 M HCl. The organic layer was dried using Na2SO4, filtered and the solvent removed in vacuo. This effectively removed triethylamine and the pure fractions were combined to give the pure product (1.22 g, 1.4 mmol, 91%) as a colourless waxy solid. 1H-NMR (400 MHz, DMSO-d6): δ = 8.06 (t, J = 5.7 Hz, 1H), 7.47-7.22 (m, 6H), 5.15 (dq, J = 8.3, 4.6 Hz, 1H), 5.01 (q, J = 12.6 Hz, 2H), 4.28 (dd, J = 12.0, 3.2 Hz, 1H), 4.11 (dd, J = 12.1, 7.0 Hz, 1H), 4.06-3.92 (m, 3H), 3.82 (q, J = 6.4 Hz, 2H), 3.33-3.18 (m, 2H), 2.27 (dt, J = 12.8, 5.0 Hz, 4H), 1.50 (q, J = 6.9 Hz, 4H), 1.32-1.16 (m, 51H), 0.85 (t, J = 6.7 Hz, 6H). 31 P-NMR (162 MHz, DMSO-d6): δ = -1.4.

[0343] Building block Ala-DPPE In a two-necked round-bottom flask, (CBz)-Ala-DPPE (308 mg, 0.34 mmol) and Pd / C (374 mg, 10% Pd, pre-wetted Degussa / Evonik type) were mixed in chloroform / ethanol 1:2 (36 mL). The flask was evacuated and refilled with Ar three times. An H2 balloon was attached, the flask was evacuated and refilled with H2 three times, and the mixture was stirred under positive H2 pressure at room temperature for 3 h. The solution was filtered through Celite and washed thoroughly with ethanol, chloroform / ethanol 1:1, and chloroform. The combined filtrate was evaporated to dryness and the resulting compound was dissolved in chloroform / ethanol 2:1 (90 mL) and dried using Na2SO4. The solution was filtered through Celite and washed thoroughly with chloroform / ethanol 2:1. The filtrate was evaporated to dryness to give the product (224 mg, 0.29 mmol, 86%) as a slightly yellowish waxy solid that contained traces of Pd. 1H-NMR (400 MHz, DMSO-d6): δ = 8.60 (br, 1H), 8.08 (br, 2H), 5.15 (br, 1H), 4.28 (d, J = 13.2 Hz, 1H), 4.12 (dd, J = 6.9 Hz, 1H), 4.00 (m, 2H), 3.88 (m, 2H), 3.80 (br, 1H), 3.09 (br, 1H), 2.35-2.23 (m, 4H), 1.50 (br, 4H), 1.41-1.14 (m, 51H), 0.85 (t, J = 6.6 Hz, 6H). 31 P-NMR (DMSO-d6): δ = -1.4.

[0344] MTP(Bn)-a-DPPE(14) MDP(Bn) (20.0 mg, 34 μmol) and Ala-DPPE (26.2 mg, 34 μmol, 1.0 equiv.) were mixed in DMAc (0.3 mL) and N,N-diisopropylethylamine (24 μL, 0.14 mmol, 4 equiv.) and PyBOP (22 mg, 41 μmol, 1.2 equiv.) was added successively. The resulting suspension was stirred at 50 °C for 1 h, after which the mixture became nearly clear. Volatiles were removed in vacuo (oil pump, 45 °C) and the mixture was flushed once with chloroform. Following column chromatography (flash SiO2) using an elution gradient of 15%-40% methanol in chloroform, this was followed by automated column chromatography (reversed-phase C18; product: C18-silica 1:200; detection: λ = 200-220 nm) using an elution gradient of 30%-80% THF in HO. This gave the product 14 (8.0 mg, 6 μmol, 18%) as a white fluffy solid after lyophilization. HPLC-MS: t[product]=3.92 min; m / z=1327.80[M+H] + (SIM mode). HPLC-ELSD: t[prod]=3.48 min; relative peak area 99.2%.

[0345] [Example 15] MTP-a-chol

[0346] [ka] Molecular weight: 1018 Daltons. CLogP=5.64.

[0347] N-(2-aminoethyl)-cholesterol carbamate building block.

[0348] [ka] A solution of cholesterol chloroformate (0.95 g, 2.1 mmol) in 20 mL of DCM was slowly added to a solution of ethylenediamine (2 mL, 14 equiv.) in 30 mL of DCM over approximately 2 h. The reaction was allowed to proceed for an additional 30 min, after which the reaction mixture was evaporated to dryness. The resulting white material was purified by column chromatography (SiO2, CHCl3 / MeOH / formic acid 78:20:2) to give 720 mg (72%) of the desired compound as a white solid. 1 H NMR (400 MHz, CDCl3) δ 5.46 - 5.27 (m, 1H), 4.99 (br. s, 1H), 4.50 (br.m, 1H), 3.22 (q, J = 5.6 Hz, 2H), 2.82 (t, J = 5.9 Hz, 2H), 2.43 - 2.19 (m, 2H), 2.06 - 1.75 (m, 5H), 1.64 - 0.80 (m, 35H), 0.68 (s, 3H) ppm. 13C NMR (101 MHz, CDCl3) δ 156.41, 139.83, 122.47, 74.31, 56.68, 56.13, 50.00, 43.63, 42.30, 41.79, 39.73, 39.51, 38.57, 36.99, 36.56, 36.18, 35.79, 31.90, 31.87, 28.22, 28.17, 28.00, 24.28, 23.82, 22.81, 22.55, 21.03, 19.33, 18.71, 11.85. m / z = 472.40 (calculated), found: 495.39 (M+Na+). A prominent peak was observed at m / z=369.37, which is attributed to the 3.4 elimination product formed in MALDI (not observed by NMR).

[0349] This building block can be coupled to N-Boc-L-alanine (CAS[15761-38-3]) via amidation; then the Boc group can be deprotected; finally, the formed amine-functional molecule can be coupled with MDP to arrive at MTP-a-chol.

[0350] [Example 16] MDP(Bn)-chol

[0351] [ka] Molecular weight: 1037 Daltons. CLogP=7.69.

[0352] The N-(2-aminoethyl)-cholesterol carbamate building block (see Example 15) can be coupled to MDP(Bn) via amidation to arrive at the molecule MDP(Bn)-chol.

[0353] [Example 17] (MTP-a-DSPE)

[0354] [Example 18] (MTP(Bn)-a-DSPE)

[0355] [Example 19] (MDP(Bn)-DSPE)

[0356] [ka] MTP-a-DSPE: MW is 1294 Daltons. CLogP=12.70 and 6.92 (uncharged and charged). MTP(Bn)-a-DSPE: MW is 1384 Daltons. CLogP=14.99 and 9.21 (uncharged and charged). MDP(Bn)-DSPE: MW is 1313 Daltons. CLogP=15.25 and 9.46 (uncharged and charged).

[0357] 1,2-Distearoyl-sn-glycero-3-phosphoethanolamine (DSPE; CAS[1069-79-0]) can be connected to N-Boc-L-alanine (CAS[15761-38-3]) via amidation; then, the Boc group can be deprotected; finally, the formed amine-functional molecule can be coupled to MDP to arrive at MTP-a-DSPE or to MDP(Bn) to arrive at MTP(Bn)-a-DSPE.

[0358] Alternatively, DSPE can be attached to MDP(Bn) via amidation to arrive at the molecule MDP(Bn)-DSPE.

[0359] [Example 20] Lipophilicity studies The ClogP values ​​of exemplary compounds of the present disclosure were evaluated using Perkin Elmer ChemDraw Professional, version 18.0.0231 (4029) software. The results show values ​​of about 4.15 to 18.28. At a physiological pH of about 7.4 (i.e., where the COOH and PO3H groups are charged), two molecules of the present invention have CLogP values ​​between 4 and 5, three have values ​​between 10 and 20, and the remaining molecule has a value between 5 and 10.

[0360] Experimentally, one can also compare the lipophilicity of molecules by running HPLC using the same elution gradient. Molecules that have a higher affinity for the hydrophobic C18 material of the column are more lipophilic and therefore have a longer retention time. The table below shows that the example molecules of the present invention (entries 1-4) have a longer retention time and are therefore more lipophilic compared to the comparative example molecules (entries 5 and 6).

[0361] Methods: HPLC-MS (SIM) and HPLC-ELSD were performed on a Phenomenex Kinetex 5 micrometer EVO C18 100A LC column (50 x 2.1 mm) using the same gradient from eluent A to B: A = 20 mM NH4HCO2 in H2O containing 0.1 v / v % formic acid, and B = 2-propanol / MeCN / H2O 85:15:5, also containing 20 mM NH4HCO2 and 0.1 v / v % formic acid.

[0362] [Table 3]

[0363] [Example 21] water solubility studies Compounds of the present disclosure were tested for solubility in PBS buffer and water applying low concentrations.

[0364] First, the compounds were weighed into vials and PBS buffer (137, 2.7, 10 and 1.8 mM in NaCl, KCl, Na2HPO4, KH2PO4, respectively, pH=7.4) was added, so that the concentration when fully dissolved was 0.2 mg / mL. The samples were shaken, left for 1 hour, shaken again and the appearance of the solution at room temperature (RT) was checked. The samples were then warmed in a 37°C water bath for 1 minute and the appearance of the solution was checked again. The results are summarized in the table below.

[0365] None of the compounds tested are spontaneously soluble in PBS at room temperature or at 37° C. In contrast, the comparative compounds tested were spontaneously soluble under these conditions. Further treatment of the sample solutions with a heat gun did not dissolve items 4, 5, and 6, whereas items 2 and 3 gave cloudy solutions after cooling to room temperature.

[0366] [Table 4]

[0367] The compound was then weighed and dissolved in chloroform / methanol. The solution was allowed to dry in the vial to form a film of material. The vial was placed under vacuum to remove traces of organic solvent. Demineralized water was added to give a compound concentration of 0.3 mM when fully dissolved (0.3 mM corresponds to 0.3 mg / mL of a compound with MW=1000 Daltons). The vial was sonicated briefly in a water bath, left overnight and sonicated again (sonication at room temperature). The appearance of the solution at room temperature was checked to assess solubility. The results are summarized in the table below.

[0368] None of the tested compounds of the present disclosure are spontaneously soluble in water at room temperature In contrast, the tested comparative compounds were spontaneously soluble under these conditions.

[0369] [Table 5]

[0370] Finally, Comparative Examples 1 and 2 were also tested for solubility in PBS (0.01M, pH=7.4) and demineralized water at a level of 1 mg / mL. The same results were found at this concentration as shown in the two tables above.

[0371] Taken together, these results show that a series of compounds of the present invention are not naturally soluble in PBS or water at concentrations as low as 0.2 mg / mL (and higher). In contrast, the comparative materials are soluble in PBS or water and give clear solutions at concentrations of at least 0.2 mg / mL and even as high as 1 mg / mL.

[0372] The disclosed compounds have low solubility in aqueous solutions, and therefore their physicochemical properties find particular use in producing stable HDL-derived NPs. Without being bound by theory, it is believed that the disclosed compounds improve fixation to NPs, reduce leakage, and provide products with better stability and shelf life.

[0373] [Example 22] Decomposition by enhanced oxidation test To achieve rapid degradation of the molecules by oxidation and mimic the slower oxidation phenomenon in vivo, the reference compounds MDP and MDP(Bn), as well as the Bn-substituted compounds from Example 7 (i.e., MDP(Bn)-DSPE [click] and Example 14 (i.e., MTP(Bn)-a-DPPE) were mixed with 12% hydrogen peroxide in water and heated at 80° C. for 4 hours.

[0374] The resulting reaction mixture is diluted with acetonitrile and water (1:1) for MDP and MDP(Bn) test solutions, or with iPrOH, acetonitrile, and water (40:7.5:52.5) ​​containing 0.1% formic acid and 20 mM ammonium formate for MDP(Bn)-DSPE[click] and MTP(Bn)-a-DPPE solutions. The four diluted samples were analyzed by HPLC-MS. As a reference, the four starting materials were also analyzed by HPLC-MS, as well as MDP-DSPE[click] and MTP-a-DPPE (i.e., debenzylated reference compounds for the Bn-substituted test molecules).

[0375] For all four test solutions, the starting compounds were traced back, which were unaffected. In addition, multiple derivatives of mass +14, +16, +28, +30, and +32 were found, indicating oxidation of CH2 to CO moieties (+14) and CH to C-OH moieties (+16), as well as combinations of these oxidation events. The tested MDP(Bn)-DSPE[click] and MTP(Bn)-a-DPPE compounds were degraded mainly by oxidation of the Bn group to benzoic acid groups (+14) followed by hydrolysis of benzoic acid (-104). This was evidenced by the presence mainly of debenzylated MDP-DSPE[click] and MTP-a-DPPE compounds as degradation products: reinforced retention times in HPLC and reinforced masses in MS (-90 compared to the starting compounds) were found.

[0376] The results show that the Bn group in the compounds of the present invention has the highest propensity for in vivo oxidative degradation. After oxidation and cleavage of Bn, a typical MDP or MTP group is formed, which degrades in vivo in a manner similar to other MDP / MTP groups known in the art.

[0377] [Example 23] Nano Biological Synthesis Method 1 - Membrane The phospholipids, (pro)drugs, and any triglycerides or polymers are dissolved (typically in chloroform, ethanol, or acetonitrile). The solution is then evaporated under vacuum to form a film of the components. A buffer solution is then added to hydrate the film and produce a vesicle suspension. The phospholipids, (pro)drugs, and any triglycerides or polymers are dissolved (typically in chloroform, ethanol, or acetonitrile). The solution is poured or added dropwise into a gently heated buffer solution with stirring until the organic solvent has completely evaporated to produce a vesicle suspension.

[0378] To the vesicle suspension generated using A or B, add apolipoprotein AI (apoA-I) (note that apoA-I can also be already present in B)-use dropwise to avoid denaturation, and sonicate the resulting mixture for 30 min using a tip sonicator while cooling completely using an external ice-water bath. Transfer the resulting solution, containing the nanobiological agent and other by-products, to a Sartorius Vivaspin tube with a molecular weight cut-off depending on the estimated size of the nanobiological agent (usually we use Vivaspin tubes with a cut-off of 10,000-100,000 kDa). Centrifuge the tube until about 90% of the solvent volume has passed through the filter. After that, add a volume of buffer approximately equal to the volume of the remaining solution and spin the tube again until almost half of the volume has passed through the filter. After repeating this twice, pass the remaining solution through a polyethersulfone 0.22 pm syringe filter to obtain the final nanobiological solution.

[0379] Method 2 – Microfluidics In an alternative approach, the phospholipids, (pro)drugs and any triglycerides, cholesterol, steryl esters, or polymers are dissolved (typically in ethanol or acetonitrile) and loaded into a syringe. Additionally, a solution of apolipoprotein AI (apoA-I) in phosphate-buffered saline is loaded into a second syringe. Using a microfluidics pump, the contents of both syringes are mixed using a microvortex platform. The resulting solution, containing the nanobiological drug and other by-products, is transferred into a Sartorius Vivaspin tube with a molecular weight cut-off depending on the estimated size of the particles (usually, Vivaspin tubes with a cut-off of 10,000-100,000 kDa are used). The tube is centrifuged until about 90% of the solvent volume has passed through the filter. After that, a volume of phosphate-buffered saline approximately equal to the volume of the remaining solution is added and the tube is spun again until almost half of the volume has passed through the filter. This is repeated twice, after which the remaining solution is passed through a polyethersulfone 0.22 pm syringe filter to obtain the final nanobiological solution.

[0380] Method 3 – Microfluidizer In another method according to the invention, the nanoscale assemblies and the final nanobiological composition are prepared using microfluidizer technology. The microfluidizer is a device for preparing materials with small particle sizes, working on the submerged jet principle. When operating the microfluidizer to obtain nanoparticles, the premix stream is passed by a high-pressure pump through a so-called interaction chamber, which consists of a channel system in a ceramic block that splits the premix into two streams. Precisely controlled shear, turbulence and cavitation forces are generated in the interaction chamber during microfluidization. The two streams recombine at high speeds, resulting in shear. The product thus obtained can be recycled back to the microfluidizer to obtain smaller and smaller particles. The advantages of microfluidization compared to traditional grinding processes include a significantly reduced contamination of the final product and easier scale-up of production.

[0381] Formulation 1 The following table provides details on the preparation of HDL-derived nanoparticle formulations. First, DMPC, cholesterol and the compound of the present invention were dissolved in ethanol (items A, B and D) or ethanol / DMSO4 / 1 (items C, E and F) at a given molar ratio, while protein apoA-1 was dissolved separately in PBS buffer (pH=7.5). In these formulations, the weight of apoA-I applied correlated with the weight of DMPC. The organic solution was mixed with PBS buffer solution by combining them by T-junction mixing.

[0382] Purification of the resulting solution was performed by TFF (tangential flow fractionation), whereby the organic solvent was removed and the nanoparticles were dissolved in PBS. Concentration of the NP solution was performed by spin filter centrifugation. Finally, the HDL-derived nanoparticle solution was filtered through a 0.2 micrometer Acrodisk PES filter.

[0383] The final HDL-derived nanoparticle solution had a typical recovery of greater than 80% for the compounds used (Examples 2, 3, 7, 8 and 13), DMPC and cholesterol. Recovery was determined using HPLC (for compounds) and assays known in the art (for DMPC and cholesterol). The concentration of the final HDL-derived nanoparticle solution was approximately 2-4 mg / mL in compounds.

[0384] [Table 6]

[0385] Stability of nanobiologics assessed by dynamic light scattering (DLS). The formulations of items A-F were characterized by DLS over 8 weeks. The nanoparticles in the formulations of Examples 2, 3, 7 and 13 had Z-average (intensity weighted average hydrodynamic size) diameters of about 20, 30, 20 and 45 nm, respectively. The dimensions of these nanoparticles remain constant over time, and the particle size distribution (PDI) also remains constant. The unloaded particles (item A) were also stable over time (diameter about 30 nm). The nanoparticles in the formulation of Example 8 showed an increase in diameter from about 50 to about 225 nm from the 2-week time point to the 5-week time point. At the 5-week time point, the dimensions were stable. Using other processing conditions, this Example 8 material can most likely also be formulated into stable 10-50 nm sized particles.

[0386] The DLS-determined Z-average diameter and PDI values ​​of the nanoparticles of formulations A–F are shown in Figure 2 .

[0387] Tip sonication formulation A: DSPC ([816-94-4]; 2.7 mg), cholesterol (0.26 mg) and compound (0.46 mg) were dissolved in a glass vial using chloroform / methanol (9:1). The solvent was removed by argon gas flow and the resulting film was dried in vacuum for >1 h. ApoA-I PBS solution (6 mL) was added to the vial followed by 5 min of sonication bath treatment, 20 min of incubation at 37 °C and then 10 min of tip sonication. The resulting dispersion was centrifuged to remove larger aggregates. The supernatant was transferred to a Vivapin20 ultrafiltration unit (cut-off 10 kDa) and spun down to a volume of approximately 1 mL. The resulting dispersion was diluted with PBS and spun down to 1 mL, and this procedure was repeated twice. Finally, the volume was diluted to 2 mL using PBS to obtain the desired nanoparticle solution.

[0388] Tip sonication formulation B: DMPC (2.7 mg), cholesterol (0.30 mg) and compound (0.57 mg) were dissolved in a glass vial using chloroform / methanol (9:1). The solvent was removed by argon gas flow and the resulting film was dried in vacuum for >1 h. A solution of peptide-2F (apoA-I mimetic 18-mer; sequence 257 in Table 2) in PBS (6 mL) was added to the vial, followed by 5 min of sonication bath treatment, 20 min of incubation at 37 °C, and then 5 min of tip sonication. The resulting dispersion was centrifuged to remove larger aggregates. The supernatant was transferred to a Vivapin20 ultrafiltration unit (cutoff 10 kDa) and spun down to approximately 1 mL. The resulting dispersion was diluted with PBS and spun down to 1 mL, and this procedure was repeated twice. Finally, the volume was diluted to 2 mL using PBS to obtain the desired nanoparticle solution.

[0389] T-junction formulation C: DMPC, cholesterol and compounds were dissolved in ethanol, while apoA-1 was dissolved in PBS buffer (pH = 7.5). T-junction mixing was applied to mix the organic solution with the buffer solution. Purification of the resulting solution was performed by TFF (tangential flow filtration), thereby removing the organic solvent. The sample was concentrated by spin-filtration. The final HDL-derived nanoparticle solution had a typical recovery of compounds, DMPC, and cholesterol of more than 75%. The concentration of the final HDL-derived nanoparticle solution was approximately 2-4 mg / mL in compounds.

[0390] [Table 7]

[0391] *apoA-I or 2F-peptide used mg per mg phosphocholine PC; **apoA-I used mg per mg compound; #number average diameter.

[0392] The above examples highlight that DSPC can be used in place of DMPC (e.g., POPC can also be used), peptidomimetics can be used in place of apoA-I, and high levels of the disclosed compounds can be incorporated. Additionally, tip sonication can be used as a processing technique, e.g., in place of T-junction mixing or microfluidic mixing.

[0393] Isolation of apolipoprotein AI (apoA-I) Human apoA-I was isolated from human HDL concentrate (Bioresource Technology) according to a previously described procedure (Zamanian-Daryoush et al., 2013). In brief, potassium bromide solution (density: 1.20 g / mL) was layered on top of the concentrate and purified HDL was obtained by ultracentrifugation. The purified fraction was defatted by adding it to a chloroform / methanol solution. The resulting milky solution was filtered and the apoA-I precipitate was dried overnight. The protein was refolded in 6 M guanidine hydrochloride and the resulting solution was dialyzed against PBS. Finally, the apoA-I PBS solution was filtered through a 0.22 pm filter and the identity and purity of the protein were confirmed by gel electrophoresis and size exclusion chromatography.

[0394] [Example 24] Cryo-TEM measurements of HDL-derived nanoparticle formulations The molecular compounds of Examples 2 and 7, respectively, namely MDP-DSPE[click] and MDP(Bn)-DSPE[click], were formulated together with 1,2-dimyristoyl-sn-glycero-3-phosphocholine (DMPC; (CAS[18194-24-6]) and APO-A1, and with various amounts of cholesterol to create HDL-derived nanoparticle formulations. The table below shows the relative molar amounts of DMPC, compound, and cholesterol components used. APO-A1 was used in twice the amount (in mg) of the compounds of the invention (in mg). Dynamic scattering data (DLS) of the processed formulations are also shown: the reported diameters and errors in brackets are based on number-averaged DLS data. The DLS recorded polydispersity in the dimensions of the particles is also shown.

[0395] [Table 8]

[0396] Formulations: DMPC, cholesterol and compounds were dissolved in ethanol (items A, C and E) or ethanol / DMSO (items B, D, F), while apoA-1 was dissolved in PBS buffer (pH=7.5). T-junction mixing was applied to mix the organic solution with the buffer solution. Purification of the resulting solution was performed by TFF (tangential flow filtration), thereby removing the organic solvent. The samples were concentrated by spin-filtration. The final HDL-derived nanoparticle solutions had typical recoveries of more than 75% for the compounds used (examples 2 and 7), DMPC and cholesterol. The recoveries were determined using HPLC (for compounds) and assays known in the art (for DMPC and cholesterol). The concentration of the final HDL-derived nanoparticle solutions was about 2-4 mg / mL in compound (examples 2 or 7).

[0397] Figure 1 shows the cryo-TEM photographs recorded for HDL-derived nanoparticles from items A-F. The 50 nm bar applies to all six photographs. When 0% cholesterol is used, spherical disk-shaped particles are mostly observed (A and B; size about 5-10 nm). When 10% cholesterol is applied, slightly expanded disks are mostly shown (C and D; length about 5-25 nm, thickness about 5 nm). When 20% cholesterol is used, clearly elongated worm-shaped particles are shown. The length of these worm-shaped particles is about 20-50 nm, and the thickness is about 5 nm (photo C). In photo F, the length of the worms is about 50-100 nm; again, the thickness is about 5 nm. In F, it is also observed that the worm-shaped particles aggregate into larger stacks.

[0398] The results highlight that the particle dimensions of HDL-derived nanoparticles can be controlled not only using the cholesterol content of the formulation (compare C with E, and D with F), but also by the lipophilicity of the compound and / or the substitution at the R2 position in formula (I) of the compounds of the invention (clear: compare E with F; less clear: in D the particles appear a bit more elongated than in C).

[0399] Methods: Immediately prior to sample processing, 200 mesh lacey carbon supported copper grids (Electron Microscopy Sciences) were surface plasma treated for 40 s using a Cressington 208 carbon coater. 3 μL of HDL-derived nanoparticle sample solution was then transferred to the grid. A thin film of the sample solution was then vitrified on the grid by plunge vitrification in liquid ethane using an automated vitrification robot (FEI Vitrobot MarkIV). The processed membranes were stored until measurements were performed. Cryo-TEM imaging of the prepared membranes was performed on a Cryo-TITAN microscope (Thermo Fisher) equipped with a field emission gun (FEG), a post-column Gatan imaging filter (model 2002), and a post-GIF 2k × 2k Gatan CCD camera (model 794).

[0400] [Example 25] In vitro NOD2 activation assay Stimulation of human NOD2 (hNOD2) by the disclosed compounds was studied by monitoring the activation of NF-κB in HEK-Blue™ hNOD2 cells (Invitrogen). 50,000 HEK-Blue™ hNOD2 cells were seeded in HEK-Blue™ detection medium in flat-bottom tissue culture plates. A range of concentrations of test articles (compounds diluted from DMSO solution (17.8 mmol / L) first in demineralized water and then in PBS to the desired concentration) were added to the cells in the tissue culture plate. The cells were incubated overnight at 37° C. with 5% CO2. The next day, the supernatant was collected into an ELISA plate and the OD was measured at 620 nm using a spectrophotometer.

[0401] The signal in this assay is based on NOD2 stimulation by ligands that subsequently activate NF-κB and AP-1, resulting in the production of SEAP. The levels of SEAP were then determined using HEK-Blue™ Detection Medium (Invitrogen). Hydrolysis of substrates in the medium by SEAP produces a purple / blue color, which was measured with an absorbance microplate reader. OD values ​​were mapped based on the concentration of the test article and are shown in Figure 3. The assay performance was validated using muramyl dipeptide (CAS number [53678-77-6]).

[0402] All of the test compounds (API) of the present invention can activate NOD2. The compounds of Examples 2, 3 and 7 have similar potency. The compounds of Examples 8 and 13 can also activate NOD2, but the extent of activation seems to be less than that of Examples 2, 3 and 7.

[0403] [Example 26] Combinatorial activity of (i) HDL-derived nanoparticles and (ii) immune checkpoint inhibition in the B16F10 mouse model 1. In Vivo Studies of Formulations Formulations for these in vivo studies were prepared by T-junction mixing followed by tangential flow filtration (TFF). Processing and purification procedures were applied as highlighted for the formulations for cryo-TEM measurements (Table 8).

[0404] The compounds used were those of Examples 2, 3, 7, 8 and 13 (i.e., these Example materials are collectively referred to as API in these descriptions of the in vivo studies). For the five formulations administered, the following relative ratios of components were used:

[0405] [Table 9]

[0406] Protocol and Results A panel of example APIs were formulated into HDL-derived nanoparticles (or nanobiologicals, NB) that were screened for their antitumor activity in combination with immune checkpoint inhibitors in a B16F10 syngeneic mouse tumor model. To this end, B16F10 mouse melanoma cells were cultured in Dulbecco's Modified Eagle Medium (DMEM) (Gibco) supplemented with 10% FBS (Gibco) and 1% penicillin / streptomycin (P / S). Cells were harvested on the day of injection and 1x10 viable cells were added to PBS containing 0.5% FBS. 6 During counting, cells were checked for viability using trypan blue solution, 0.4% (Gibco). At the start of the experiment, 1 × 10 cells were resuspended in 100 μL PBS supplemented with 0.5% fetal bovine serum (FBS). 5 B16F10 tumor cells were injected subcutaneously into the flank of 7-week-old female C57BL / 6 mice (The Jackson Laboratory).

[0407] Seven days after tumor inoculation, mice were randomized into groups with similar average group size (n=10). The average tumor size for the groups was 3.26 mm 3 After randomization, mice were ear-nicked and weighed. The dose was then calculated and aliquoted. The aliquoted doses were stored at 4° C. until use.

[0408] The study consisted of the following: a PBS control group, an immune checkpoint inhibitor group (CI) and six treatment groups. Mice treated with immune checkpoint inhibitors received intraperitoneal injections using a dose of 200 μg anti-CTLA-4 (clone, 9H10, BioXcell) and / or 200 μg anti-PD-1 (clone, RMP1-14, BioXcell) on days 2, 4, and 8. Treatment groups consisted of NB prepared from the compound of Example 2, NB from Example 3, NB from Example 7, NB from Example 8, and NB from Example 13 in combination with immune checkpoint inhibitor therapy (CI) as described above. The dose of the treatment groups was about 9 mg MDP / kg (or about 27 mg / kg of each API contained in the NB) on days 0, 2, and 4.

[0409] Tumor growth curves are shown in Figure 4A-E, with the same PBS and CI groups mapped in each graph, allowing comparison between graphs.

[0410] PBS-treated animals or animals treated with immune checkpoint inhibitors alone did not show tumor growth inhibition. All groups of animals treated with combination therapy showed tumor growth inhibition, which was most pronounced in groups in which NB of Example 2 (FIG. 4A), NB of Example 7 (FIG. 4C) or NB of Example 8 (FIG. 4D) were part of the combination therapy. It should be noted that the above results regarding the combination therapy were obtained with the applied formulation containing 10 mol% cholesterol for the applied 90 mol% DMPC, and therefore the dimensions of these particles were approximately 5 up to 10 nm (see cryo-TEM panels C and D in Figure 1) (see table above).

[0411] [Example 27] Single-agent activity of nanobiologics in the B16F10 mouse model Formulations for these in vivo studies were prepared by T-junction mixing followed by tangential flow filtration (TFF). Processing and purification procedures were applied as highlighted in the formulations for cryo-TEM measurements (Table 8). The table below shows the relative ratios of the components used to prepare the HDL-derived nanoparticles.

[0412] [Table 10]

[0413] Protocol and Results Two APIs (compounds of Example 2 and Example 7) were used to generate a series of different HDL-derived nanoparticle formulations (or nanobiological preparations; nanobiological preparations; NBs) and determine their potency. The resulting nanobiological preparations were screened for single-agent antitumor activity in the B16F10 syngeneic mouse tumor model. To this end, B16F10 mouse melanoma cells were cultured in Dulbecco's Modified Eagle's Medium (DMEM) (Gibco) supplemented with 10% FBS (Gibco) and 1% penicillin / streptomycin (P / S). Cells were harvested on the day of injection and 1 × 10 viable cells were added to PBS containing 0.5% FBS. 6 Cells were resuspended at 1 × 10 cells / mL. During counting, cell viability was checked using a cell counter and analyzer (Casy). At the start of the experiment, 1 × 10 cells were resuspended in 100 μL PBS supplemented with 0.5% fetal bovine serum (FBS). 5 B16F10 tumor cells were injected subcutaneously into the flank of 7-week-old female C57BL / 6J mice (Charles River).

[0414] Seven days after tumor inoculation, mice were randomized into groups with similar average group size. Groups consisted of 8-10 mice. The average tumor size for the group was 6.33 mm. 3 After randomization, mice were tattooed with numbers on their tails. The doses were then calculated and aliquoted. The aliquots were stored at 4°C until use.

[0415] The study consisted of: a PBS control group, and five treatment groups: NB of Example 2 (formulation items 1, 3 and 4) and NB of Example 7 (formulation items 2 and 5). The doses of the treatment groups were about 3 mg MDP / kg (or about 9 mg / kg of the respective API in the NB) on days 0, 2 and 4. Tumor size was measured at set times during the study. The tumor growth curves are shown in Figures 5A-C, mapping the same PBS group in each graph to allow comparison between the graphs. Apart from NB of Example 2 (formulation item 1), all nanobiological formulations clearly showed a reduction in tumor growth compared to the PBS control group. For both Example 2 (Figure 5A) and Example 7 (Figure 5B), the nanobiological formulation with 20 mol% cholesterol performed the best. Figure 5C shows that NB of Example 2 is slightly better than NB of Example 7 (using formulation items 4 and 5, respectively).

[0416] In Figure 5A, the formulation of Example 2 containing 10 mol% cholesterol (item 1 in the table above) showed only slight single-agent activity (compared to PBS). Surprisingly, in comparison, the formulations of Example 2 containing 20 mol% cholesterol (items 3 and 4 in the table above) show highly enhanced tumor inhibition. This can also be seen for the formulation of Example 7 (Figure 5B). Clearly, the dimensions of the formed particles play a decisive role in the activity of the prepared HDL-derived nanoparticles: the 10% cholesterol formulation gives spherical or slightly elongated discoidal particles with dimensions of 5-10 nm, while the 20% cholesterol formulation gives elongated worm-like particles with lengths of about 15-50 nm and thickness of about 5 nm (compare panels C and E in Figure 1, cryo-TEM data).

[0417] The disclosed HDL-derived nanoparticles have certain dimensions, and these characteristics find particular use in generating stable and potent HDL-derived NPs. Without being bound by theory, it is believed that the disclosed HDL-derived nanoparticles improve the (multivalent) presentation of MDP (or MDP(Bn) or MTP or MTP(Bn)) moieties to cells, dramatically improving their efficacy.

[0418] Embodiment 1. Formula (I):

[0419] [ka] or a pharma- ceutically acceptable salt thereof. (In the formula, R 1 is -H or -C(O)-R X and; R 2 and R 3 are each independently selected from the group consisting of -H, alkyl, alkylene-aryl, -C(O)-alkyl, and -C(O)-aryl; R 4 , R 5 , and R 5’ are each alkyl; R 6 and R 11 are each independently -H or alkyl; R 7 is C 9~30 Fatty acid chain, -YN(R 11 )-C(O)-O-alkylene-C(H)(OR 8 )-Alkylene-OR 9 , -C(R 10 )(C(O)NH2)-alkylene-N(R 11 )-C(O)-C 16~30 Fatty acid chain, -(CR 10 R 10 )2-OP(O)(OH)-O-alkylene-C(R 10 )(OR Z )-Alkylene-OR Zor -Y-triazolyl-L; R Z is C 8~30 Fatty acids or -C(O)-C 16~30 is a fatty acid chain; Y is alkylene; L is a fatty acid chain, -alkylene-C(O)-W, -alkylene-OC(O)-W, -alkylene-N-(alkylene-C(O)-NR 11 -Alkylene-NR 11 -C(O)-W)2, and -alkylene-N-(alkylene-C(O)-W)2; W is a fatty acid chain, -O-alkylene-C(H)(OR 8 )-Alkylene-OR 9 , a phospholipid, or a sterol; R 8 and R 9 are each independently R X or -C(O)-R X and; R 10 , R 22 , R 33 , R 33’ , R 44 , R 44’ , R 55 , and R 55’ are each independently H or R A and; R X is a fatty acid chain; Each of the foregoing alkyl, alkylene, alkylene-aryl, aryl, and triazolyl may be selected from one or more R A Optionally replaced by R A is halo, alkoxy, haloalkoxy, cyano, hydroxyl, -N(R C )(R D ), -C(O)N(R C )(R D ), -N(R C )C(O)R B , -OC(O)NR C R D , -NR C C(O)OR B , -OC(O)RB , -C(O)OR B , -C(O)R B , -CO2H, -NO2, -SH, S(O) X R B (wherein X is 0, 1, or 2), aryl, arylalkyl, heteroaryl, heteroarylalkyl, cycloalkyl, heterocycloalkyl, and R B independently selected for each occurrence from the group consisting of: R C and R D is hydrogen, alkyl, haloalkyl-C(O)R B , and -C(O)OR B or R C and R D together with the nitrogen to which they are attached, R A forming an optionally substituted heterocycle; R B is alkyl, alkenyl, or alkynyl optionally substituted with one or more fluoro; R 7 C 9~30 If it is a fatty acid chain, R 2 is -H).

[0420] 2. The compound of formula (I) has the formula (IA):

[0421] [ka] or a pharma- ceutically acceptable salt thereof.

[0422] 3. R 2 The compound of embodiment 1 or 2, wherein is -H or benzyl.

[0423] 4. R 2 The compound of any one of embodiments 1-3, wherein is -H.

[0424] 5. R 10 , R 22 , R33 , R 33’ , R 44 , R 44’ , R 55 , and R 55’ The compound of any one of embodiments 1-4, wherein each is -H.

[0425] 6. R 4 The compound of any one of embodiments 1-5, wherein is alkyl.

[0426] 7. R 4 The compound of any one of embodiments 1-6, wherein is methyl.

[0427] 8. R 3 and R 6 The compound of any one of embodiments 1-7, wherein:

[0428] 9. Y is -C(O)N(R C )(R D 9. The compound of any one of embodiments 1-8, wherein R is an alkylene optionally substituted with R.

[0429] 10. Y is -CH2- or

[0430] [ka] 10. The compound of embodiment 9, wherein

[0431] 11. The compound of embodiment 10, wherein Y is -CH2-.

[0432] 12. R 7 The compound of any one of embodiments 1-11, wherein is -Y-triazolyl-L.

[0433] 13. The compound of formula (I) is a compound of formula (II):

[0434] [ka] or a pharma- ceutically acceptable salt thereof. (In the formula, X1 is -N- and X2 is -C-; or X1 is -C- and X2 is -N-. 13. The compound of any one of embodiments 1-12, wherein

[0435] 14. The compound of formula (I) has the formula (IIA):

[0436] [ka] or a pharma- ceutically acceptable salt thereof.

[0437] 15. Y is C 1~6 The compound of embodiment 13 or 14, wherein the compound is alkylene.

[0438] 16. R A The compound of any one of embodiments 13-15, wherein is -H.

[0439] 17. L is C 8~30 Fatty acid chain, -CH2-C(O)-W, -CH2-OC(O)-W, -CH2CH2-N-CH2CH2-C(O)-NR 11 -CH2CH2-NR 11 The compound according to any one of the preceding embodiments, wherein the compound is selected from the group consisting of: -C(O)-W)2, -CH2CH2-N-(CH2CH2-C(O)-W)2, and -CH2CH2-N-(CH2CH2-C(O)-W)2.

[0440] 18. L is C 12~18 The compound of embodiment 17, which is a fatty acid chain.

[0441] 19. The compound of embodiment 17, wherein L is -CH2(CH2CH2)8-CH3.

[0442] 20. W is C 8~30 The compound according to any one of embodiments 1 to 19, which is a fatty acid chain.

[0443] 21. W is C 12~18 The compound according to any one of embodiments 1 to 19, which is a fatty acid chain.

[0444] 22. W,

[0445] [ka] 20. The compound of any one of embodiments 1-19, wherein

[0446] 23. R X and R X’ However, each independently -C 8~30 The compound of embodiment 22, which is a fatty acid chain.

[0447] 24. R X and R X’ However, each is independently C 12~18 The compound according to embodiment 22 or 23, which is a fatty acid chain.

[0448] 25. R X and R X’ are both -(CH2CH2)8-CH3.

[0449] 26. W is cholesterol:

[0450] [ka] 20. The compound of any one of embodiments 1-19, wherein

[0451] 27. The compound of any one of embodiments 1-19, wherein W is a phospholipid selected from the group consisting of phosphatidylcholine (PC), phosphatidylglycerol (PG), phosphatidylserine (PS), phosphatidylethanolamine (PE), phosphatidic acid (PA), and lysophosphatidylcholine.

[0452] 28. W is structure:

[0453] [ka] or a pharma- ceutically acceptable salt thereof. (In the formula, R X and R X’ are each independently C 8~30 fatty acid chain)

[0454] 29. R X and R X’ However, each is independently C 12~18 The compound of embodiment 28, which is a fatty acid.

[0455] 30. The compound of embodiment 28 or 29, wherein the fatty acid is saturated.

[0456] 31. R X and R X’ are both -(CH2CH2)8-CH3.

[0457] 32. R 7 is -C(H)(C(O)NH2)-C5 alkylene-N(R 11 )-C(O)-C 17~30 The compound of any one of embodiments 1-11, which is a fatty acid.

[0458] 33. R 7 is -CH2CH2-OP(O)(OH)-O-CH2-C(H)(OR Z )-CH2-OR Z 12. The compound of any one of embodiments 1-11, wherein

[0459] 34.

[0460] [ka]

[0461] [ka]

[0462] [ka]

[0463] [ka] The compound according to any one of embodiments 1 to 33, selected from the group consisting of:

[0464] 35. A nanobiological composition comprising high density lipoprotein (HDL)-derived nanoparticles, the nanoparticles having formula (I):

[0465] [ka] or a pharma- ceutically acceptable salt thereof (In the formula, R 1 is -H or -C(O)-R X and; R 2 and R 3 are each independently selected from the group consisting of -H, alkyl, alkylene-aryl, -C(O)-alkyl, and -C(O)-aryl; R 4 , R 5 , and R 5’ are each alkyl; R 6 and R 11 are each independently -H or alkyl; R 7 is the fatty acid chain, -YN(R 6 )-C(O)-O-alkylene-C(H)(OR 8 )-Alkylene-OR 9 , -YN(R 6 )-C(O)-R X , -YOP(O)(OH)-O-alkylene-C(H)(OR8 )-Alkylene-OR 9 or -Y-triazolyl-L; Y is alkylene; L is a fatty acid chain, -alkylene-C(O)-W, -alkylene-OC(O)-W, -alkylene-N-(alkylene-C(O)-NR 11 -Alkylene-NR 11 -C(O)-W)2, and -alkylene-N-(alkylene-C(O)-W)2; W is a fatty acid chain, -O-alkylene-C(H)(OR 8 )-Alkylene-OR 9 , a phospholipid, or a sterol; R 8 and R 9 are each independently R X or -C(O)-R X and; R 10 , R 22 , R 33 , R 33’ , R 44 , R 44’ , R 55 , and R 55’ are each independently H or R A and; R X is a fatty acid chain; Each of the foregoing alkyl, alkylene, alkylene-aryl, aryl, and triazolyl may be selected from one or more R A Optionally replaced by R A is halo, alkoxy, haloalkoxy, cyano, hydroxyl, -N(R C )(R D ), -C(O)N(R C )(R D ), -N(R C )C(O)R B , -OC(O)NR C R D , -NR C C(O)OR B , -OC(O)R B , -C(O)OR B , -C(O)R B, -CO2H, -NO2, -SH, S(O) X R B (wherein X is 0, 1, or 2), aryl, heteroaryl, heteroarylalkyl, cycloalkyl, heterocycloalkyl, and R B independently selected for each occurrence from the group consisting of: R C and R D is hydrogen, alkyl, haloalkyl-C(O)R B , and -C(O)OR B or R C and R D together with the nitrogen to which they are attached, R A forming an optionally substituted heterocycle; R B is alkyl, alkenyl, or alkynyl optionally substituted with one or more fluoro. 13. A nanobiological composition comprising:

[0466] 36. A nanobiological composition comprising high density lipoprotein (HDL)-derived nanoparticles, the nanoparticles comprising a compound according to any one of embodiments 1 to 34.

[0467] 37. The nanobiological composition of embodiment 35 or 36, wherein the HDL-derived nanoparticles comprise one or more phospholipids.

[0468] 38. The nanobiological composition of embodiment 37, wherein the phospholipid is independently selected from the group consisting of phosphatidylcholine, phosphatidylethanolamine, phosphatidylinositol, phosphatidylserine, sphingomyelin or other ceramides, phospholipid-containing oils, phosphatidylglycerol, phosphatidic acid, lysophosphatidylcholine, and combinations thereof.

[0469] 39. The nanobiological composition according to embodiment 35 or 36, comprising a phospholipid selected from the group consisting of 1,2-dimyristoyl-sn-glycero-3-phosphatidylcholine (DMPC) and 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC), and a lysolipid selected from the group consisting of 1-myristoyl-2-hydroxy-sn-glycero-phosphocholine (MHPC), and 1-palmitoyl-2-hydroxy-sn-glycero-3-phosphocholine (PHPC).

[0470] 40. The nanobiological composition of any one of embodiments 35 to 39, wherein the HDL-derived nanoparticles comprise apoA-I or a peptidomimetic of apoA-I.

[0471] 41. The nanobiological composition of any one of embodiments 35 to 40, wherein the HDL-derived nanoparticles further comprise one or more triglycerides, fatty acid esters, hydrophobic polymers, sterol esters, or combinations thereof.

[0472] 42. The nanobiological composition of any one of embodiments 35-41, wherein the HDL-derived nanoparticles further comprise cholesterol.

[0473] 43. The nanobiological composition of embodiment 42, wherein the HDL-derived nanoparticles comprise one or more phospholipids and cholesterol in a molar ratio ranging from about 1:0.05 to about 1:0.25.

[0474] 44. The nanobiological composition of embodiment 43, wherein the HDL-derived nanoparticles comprise one or more phospholipids and cholesterol in a molar ratio ranging from about 1:0.2.

[0475] 45. A nanobiological composition according to any one of embodiments 1 to 44, wherein the HDL-derived nanoparticles are nanodiscs or nanospheres.

[0476] 46. ​​The nanobiological composition of embodiment 45, wherein the nanodisk or nanosphere has a diameter of about 8 nm to about 400 nm.

[0477] 47. A pharmaceutical composition comprising a compound according to any one of embodiments 1 to 34, or a pharma- ceutically acceptable salt thereof, and a pharma- ceutically acceptable carrier.

[0478] 48. A method for treating a cell proliferation disorder in a patient in need thereof, comprising administering to the patient a therapeutically effective amount of a nanobiological composition described in any one of embodiments 35-46.

[0479] 49. The method of embodiment 48, wherein the cell proliferation disorder is cancer.

[0480] 50. The method of embodiment 49, wherein the cancer is selected from the group consisting of bladder cancer, vascular cancer, bone cancer, brain cancer, breast cancer, cervical cancer, breast cancer, colon cancer, endometrial cancer, esophageal cancer, eye cancer, head cancer, kidney cancer, liver cancer, lymph node cancer, lung cancer, oral cancer, cervical cancer, ovarian cancer, pancreatic cancer, prostate cancer, rectal cancer, colorectal cancer, skin cancer, stomach cancer, testicular cancer, pharyngeal cancer, thyroid cancer, urothelial cancer, and uterine cancer.

[0481] 51. The method of embodiment 49, wherein the cancer is selected from the group consisting of breast cancer, prostate cancer, melanoma, colorectal cancer, lung cancer, pancreatic cancer, and glioblastoma.

[0482] 52. The method according to any one of embodiments 48-51, further comprising co-administering an anti-cancer drug with the nanobiological composition as a combination therapy.

[0483] 53. A method for treating sepsis in a patient in need thereof, comprising administering to the patient a therapeutically effective amount of a nanobiological composition described in any one of embodiments 35-46.

[0484] 54. The method of embodiment 53, wherein the patient has sepsis associated with a bacterial, viral or fungal infection of the lungs, abdomen, kidneys, or bloodstream.

[0485] 55. The method of any one of embodiments 48-54, wherein the nanobiological composition promotes a hyper-reactive innate immune response in a patient in need thereof.

[0486] 56. The method of embodiment 55, wherein the hyperreactive innate immune response is promoted for at least about 7 to about 30 days.

[0487] 57. The method of embodiment 55, wherein the hyperreactive innate immune response is promoted for at least 30 to 100 days.

[0488] 58. The method of embodiment 55, wherein the hyperreactive innate immune response is promoted for more than 100 days and up to 3 years.

[0489] 59. The method of embodiment 55, wherein the nanobiological composition is administered once and the hyper-reactive innate immune response is promoted for at least 30 days.

[0490] 60. The method of embodiment 55, wherein the nanobiological composition is administered at least once a day on each day of a multiple dose regimen, and the hyper-reactive innate immune response is promoted for at least 30 days.

[0491] 61. The method of any one of embodiments 48-58, wherein the nanobiological composition is administered to the patient in a treatment regimen comprising two or more doses to generate accumulation of the drug in bone marrow cells, bone marrow progenitor cells, and hematopoietic stem cells in the bone marrow, blood, and / or spleen.

[0492] 62. The method of any one of embodiments 48-61, wherein the nanobiological composition is administered intravenously or intraarterially.

[0493] 63. A method for activating a NOD2 receptor in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a nanobiological composition described in any one of embodiments 35 to 46.

[0494] 64. A process for producing a nanobiological composition according to any one of embodiments 35 to 46, comprising: a) forming, under conditions effective to form a lipid membrane, a lipid membrane comprising: i) a compound according to any one of embodiments 1-34; ii) one or more phospholipids; optionally, iii) a hydrophobic matrix comprising one or more triglycerides, fatty acid esters, hydrophobic polymers, or sterol esters, or combinations thereof; and optionally iv) cholesterol; and b) dissolving the lipid membrane in a solvent to form a lipid solution; and contacting the lipid solution with apoA-I or a peptide mimetic of apoA-I under conditions effective to form HDL-derived nanoparticles comprising a compound described in any one of embodiments 1 to 34.

[0495] 65. A nanobiological composition prepared according to embodiment 64.

[0496] 66. A kit comprising a nanobiological composition according to any one of embodiments 35 to 46.

[0497] 67. The method of claim 52, wherein the anticancer drug is a checkpoint inhibitor.

[0498] 68. The method of claim 67, wherein the checkpoint inhibitor is selected from an anti-PD-1 antibody, an anti-CTLA-4 antibody, and a combination thereof.

Claims

1. Formula (I): 【Chemistry 1】 or a pharma- ceutically acceptable salt thereof. (In the formula, R １ is -H or -C(O)-R Ｘ and R ２ and R ３ are each independently selected from the group consisting of -H, alkyl, aryl, alkylene-aryl, -C(O)-alkyl, and -C(O)-aryl; R ４ , R ５ , and R ５’ are each alkyl; R ６ and R １１ are each independently —H or alkyl; R ７ is -Y-triazolyl-L; R Ｚ is C ８～３０ Fatty acid chain or -C(O)-C １６～３０ is a fatty acid chain; Y is alkylene; L is a fatty acid chain, -alkylene-C(O)-W, -alkylene-O-C(O)-W, -alkylene-N-(alkylene-C(O)-NR １１ -Alkylene-NR １１ -C(O)-W) ２ and -alkylene-N-(alkylene-C(O)-W) ２ selected from the group consisting of: W is a fatty acid chain, -O-alkylene-C(H)(OR ８ )-alkylene-OR ９ , a phospholipid, or a sterol; R ８ and R ９ are each independently R Ｘ or -C(O)-R Ｘ and R ２２ , R ３３ , R ３３’ , R ４４ , R ４４’ , R ５５ , and R ５５’ are each independently H or R Ａ and R Ｘ is a fatty acid chain; Each of the foregoing alkyl, alkylene, alkylene-aryl, aryl, and triazolyl may be selected from one or more R Ａ and optionally substituted with R Ａ is hydrogen, halo, alkoxy, haloalkoxy, cyano, hydroxyl, -N(R Ｃ ) (R Ｄ ), -C(O)N(R Ｃ ) (R Ｄ ), -N(R Ｃ ) C(O)R Ｂ , -OC(O)NR Ｃ R Ｄ , -NR Ｃ C(O)OR Ｂ , -OC(O)R Ｂ , -C(O)OR Ｂ , -C(O)R Ｂ , -CO ２ H, -NO ２ , -SH,S(O) Ｘ R Ｂ (wherein X is 0, 1, or 2), aryl, arylalkyl, heteroaryl, heteroarylalkyl, cycloalkyl, heterocycloalkyl, and R Ｂ independently selected for each occurrence from the group consisting of: R Ｃ and R Ｄ is hydrogen, alkyl, haloalkyl-C(O)R Ｂ and -C(O)OR Ｂ or R Ｃ and R Ｄ together with the nitrogen to which they are attached, R Ａ forming an optionally substituted heterocycle; R Ｂ is alkyl, alkenyl, or alkynyl optionally substituted with one or more fluoro; R ７ C ９～３０ When it is a fatty acid chain, R ２ is -H).

2. The compound of formula (I) has the formula (IA): 【Chemistry 2】 2. The compound of claim 1, which is a compound of formula (I) or a pharma- ceutically acceptable salt thereof.

3. R ２ The compound of claim 1, wherein is -H or benzyl.

4. R ２ The compound of claim 1 , wherein is —H.

5. R ２２ , R ３３ , R ３３’ , R ４４ , R ４４’ , R ５５ , and R ５５’ The compound of claim 1 , wherein each is —H.

6. R ４ The compound of claim 1 , wherein is alkyl.

7. R ４ The compound of claim 4 , wherein is methyl.

8. R ３ and R ６ The compound of claim 1 , wherein both are —H.

9. Y is -CH ２ - 2. The compound of claim 1,

10. The compound of formula (I) is represented by formula (II): 【Chemistry 3】 or a pharma- ceutically acceptable salt thereof. (In the formula, X １ is -N-, and X ２ is -C-; or X １ is -C-, and X ２ is -N-) 2. The compound of claim 1,

11. The compound of formula (II) has the formula (IIA): 【Chemistry 4】 or a pharma- ceutically acceptable salt thereof.

12. Y is C １～６ The compound of claim 10 which is an alkylene.

13. R Ａ The compound of claim 10, wherein is -H.

14. L is -CH ２ The compound of claim 1 which is -C(O)-W.

15. W is C ８～３０ The compound of claim 1 , wherein the aryl group is an alkyl group.

16. W, 【Chemistry 5】 (In the formula, R Ｘ and R Ｘ’ are each independently -C ８～３０ fatty acid chain) 2. The compound of claim 1,

17. R Ｘ and R Ｘ’ However, each independently -C ８～３０ 17. The compound of claim 16, which is alkyl.

18. R Ｘ and R Ｘ’ But both are -(CH ２ CH ２ ) ８ -CH ３ 18. The compound of claim 17,

19. W is cholesterol: 【Chemistry 6】 2. The compound of claim 1,

20. The compound of claim 1 , wherein W is a phospholipid.

21. W has the structure: 【Chemistry 7】 Phospholipids having (In the formula, Y Ｑ１ , Y Ｑ２ , and Y Ｑ３ are each independently alkylene; R Ｘ and R Ｘ’ are each independently a fatty acid chain having at least 15 carbons.

21. The compound of claim 20,

22. R Ｘ and R Ｘ’ However, each independently C １５～２０ Alkyl or C １５～２０ 22. The compound of claim 21 which is alkenyl.

23. W has the structure: 【Chemistry 8】 or a pharma- ceutically acceptable salt thereof. (In the formula, R Ｘ and R Ｘ’ are each independently C ８～３０ fatty acid chain) 22. The compound of claim 21 ,

24. R Ｘ and R Ｘ’ However, each independently C １２～１８ 24. The compound of claim 23 which is a fatty acid.

25. 24. The compound of claim 23, wherein the fatty acid is saturated.

26. R Ｘ and R Ｘ’ But both are -(CH ２ CH ２ ) ８ -CH ３ 24. The compound of claim 23, wherein 【Request 27】 【Chemical 9-1】 【Chemistry 9-2】 【Chemistry 9-3】 and pharma- ceutically acceptable salts thereof, wherein Bn is a benzyl group.

28. 13. A nanobiological composition comprising high density lipoprotein (HDL)-derived nanoparticles, said nanoparticles comprising one or more phospholipids and the compound of claim 1.

29. 29. The nanobiological composition of claim 28, wherein said phospholipids are independently selected from the group consisting of phosphatidylcholine, phosphatidylethanolamine, phosphatidylinositol, phosphatidylserine, sphingomyelin or other ceramides, phospholipid-containing oils, phosphatidylglycerol, phosphatidic acid, lysophosphatidylcholine, and combinations thereof.

30. 29. The nanobiological composition of claim 28, further comprising a phospholipid selected from the group consisting of 1,2-dimyristoyl-sn-glycero-3-phosphatidylcholine (DMPC) and 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC), and a lysolipid selected from the group consisting of 1-myristoyl-2-hydroxy-sn-glycero-phosphocholine (MHPC), and 1-palmitoyl-2-hydroxy-sn-glycero-3-phosphocholine (PHPC).

31. 30. The nanobiological composition of claim 28, wherein said HDL-derived nanoparticles comprise apoA-I or a peptidomimetic of apoA-I.

32. 30. The nanobiological composition of claim 28, wherein said HDL-derived nanoparticles further comprise cholesterol.

33. 33. The nanobiological composition of claim 32, wherein said HDL-derived nanoparticles comprise one or more phospholipids and cholesterol in a molar ratio ranging from about 1:0.05 to about 1:0.

25.

34. 10. A pharmaceutical composition comprising a compound of claim 1 or a pharma- ceutically acceptable salt thereof, and a pharma- ceutically acceptable carrier.

35. 30. A composition for activating the NOD2 receptor, comprising a therapeutically effective amount of the nanobiological composition of claim 28.

36. The compound of formula (II) has the formula (II-I): 【Chemistry 10】 or a pharma- ceutically acceptable salt thereof.

37. The compound is 【Chemistry 11】 28. The compound of claim 27, which is a compound of the formula:

38. The compound is 【Chemistry 12】 28. The compound of claim 27, which is a compound of the formula:

39. The compound is 【Chemistry 13】 28. The compound of claim 27, which is a compound of the formula:

40. The compound is 【Chemistry 14】 28. The compound of claim 27, which is a compound of the formula:

Images