Compounds for modulating trained immunity and their methods of use

Abstract

This invention relates to compounds for modulating training immunity and methods of using the same. This document provides compounds of formula (I), compositions comprising compounds of formula (I), and their uses.

Description

[0001] This application is a divisional application of patent application No. 202280035822.6 (International Application No. PCT / US2022 / 021035), filed on March 18, 2022, entitled "Compounds for Regulating Training Immunity and Methods of Using Them Thereof".

[0002] Cross-reference to related applications

[0003] This application claims priority to U.S. Provisional Application No. 63 / 163,428, filed March 19, 2021, the disclosure of which is incorporated herein by reference in its entirety for all purposes.

[0004] Description of the text file submitted electronically

[0005] The contents of the text file submitted electronically here are incorporated herein by reference in their entirety: a computer-readable copy of the sequence list (filename: TRAI_005_00US_SeqList_ST25.txt, date of record: March 17, 2021, file size 105 kilobytes). Background Technology

[0006] The immune system plays a crucial role in the pathophysiology of major diseases such as atherosclerosis, diabetes, and cancer. However, most immunotherapeutic strategies currently under development focus on effector molecules such as cytokines or T lymphocytes as cells derived 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, anti-cytokine therapy can successfully neutralize bioactive cytokines, while the most commonly used immunotherapies in cancer patients include the use of checkpoint inhibitor drugs. Although the innate immune system has long been thought to lack memory, recent studies have shown that innate immune cells undergo metabolic and epigenetic rewiring, adjusting their functional programs in a process called 'trained immunity', which is related to their anti-tumor effects (Buffen et al., 2014, PLoSPathog. 10, e1004485; Netea et al., J. Leukoc Biol. 2017, 102, 1323-1332).

[0007] A range of pattern recognition receptors (PRRs), including TLRs, NOD2, dectin 1, and inflammasomes, can participate in promoting training immunity. Furthermore, in vitro studies have demonstrated the therapeutic potential of BCG and several other PAMPs and DAMPs (including peptidoglycan and β-glucan) as agents that promote training immunity. However, the in vivo therapeutic use of molecules that regulate training immunity is hampered by toxicity, immune-related adverse effects, and poor bioavailability targeting relevant myeloid cells and their progenitors.

[0008] There is a need for therapeutic agents and combinations thereof that participate in the innate immune system and regulate trained immunity to treat cancer and other diseases and conditions caused by defective trained immunity. Summary of the Invention

[0009] In the implementation scheme, compounds of formula (I) or pharmaceutically acceptable salts thereof are provided herein:

[0010]

[0011] in:

[0012] R 1 -H or -C(O)-R X ;

[0013] R 2 and R 3 Each is independently selected from the group consisting of -H, alkyl, alkylene-aryl, -C(O)-alkyl, and -C(O)-aryl;

[0014] R 4 R 5 and R 5’ Each is an alkyl group;

[0015] R 6 and R 11 Each is independently -H or alkyl;

[0016] R 7 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 -γ-triazolyl-L;

[0017] R Z and R Z’ Each independently is C 8-30 Fatty acids or -C(O)-C 16-30 fatty acid chains;

[0018] Y is an alkylene group;

[0019] L is selected from fatty acid chains, -alkylene-C(O)-W, -alkylene-OC(O)-W, -alkylene-N-(alkylene-C(O)-NR) 11 -alkylene-NR 11 The group consisting of -C(O)-W)2 and -alkylene-N-(alkylene-C(O)-W)2;

[0020] W represents a fatty acid chain, -O-alkylene-C(H)(OR) 8 )-alkylene-OR 9 Phospholipids or sterols;

[0021] R 8 and R 9 Each independently as R X or -C(O)-R X ;

[0022] R 10 R 22 R 33 R 33’ R 44 R 44’ R 55 and R 55’ Each independently is H or R A ;

[0023] R X It consists of fatty acid chains;

[0024] Each of the aforementioned alkyl, alkylene, alkylene-aryl, aryl, and triazolyl groups is optionally surrounded by one or more R groups. A Replace, where R A Each time it appears, it is independently selected from the following groups: hydrogen, halogen, 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 -NRC C(O)OR B -OC(O)R B -C(O)OR B -C(O)R B -CO2H, -NO2, -SH, S(O) X R B (where X is 0, 1, or 2), aryl, arylalkyl, heteroaryl, heteroarylalkyl, cycloalkyl, heterocycloalkyl, and R B ;

[0025] R C and R D Each occurrence is independently selected from hydrogen, alkyl, haloalkyl, -C(O)R B and -C(O)OR B The group formed; or R C and R D Together with the nitrogen they are attached to, they form optional R A Substituted heterocycles; and

[0026] R B It can be an alkyl, alkenyl, or alkynyl group optionally substituted with one or more fluorine groups;

[0027] Where R 7 C 9-30 When fatty acid chains are in the form of R 2 It is -H.

[0028] In the implementation scheme, the compound of formula (I) is the compound of formula (IA) or a pharmaceutically acceptable salt thereof:

[0029]

[0030] Where 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 Defined in this article.

[0031] In the implementation scheme, the compound of formula (I) is the same as the compound of formula (IB):

[0032]

[0033] Where R2 R 3 R 4 R 6 and R 7 Defined in this article.

[0034] In the implementation scheme, the compound of formula (I) is the compound of formula (II) or a pharmaceutically acceptable salt thereof:

[0035]

[0036] Where R 2 R 3 R 4 R 5 R 5’ R 6 Y, X 1 X 2 R A L is defined in this paper.

[0037] In the implementation scheme, the compound of formula (II) is:

[0038]

[0039] Or a pharmaceutically acceptable salt thereof, wherein R 2 R 3 R 4 R 5 R 5’ R 6 Y, X 1 X 2 L is defined in this paper.

[0040] In the implementation scheme, the compound of formula (I) is a compound of formula (IIA) or a pharmaceutically acceptable salt thereof:

[0041]

[0042] Where R 2 R 3 R 4 R 5 R 5’ R 6 Y, X 1 X 2 L and R A Defined in this article.

[0043] In the implementation scheme, the compound of formula (IIA) is:

[0044]

[0045] Where R2 R 3 R 4 R 5 R 5’ R 6 Y, X 1 X 2 L is defined in this paper.

[0046] In its embodiments, this disclosure provides nanobiocompositions comprising nanoparticles derived from high-density lipoprotein (HDL), wherein the nanoparticles comprise a compound of formula (I) or a pharmaceutically acceptable salt thereof:

[0047]

[0048] in:

[0049] R 1 -H or -C(O)-R X ;

[0050] R 2 and R 3 Each is independently selected from the group consisting of -H, alkyl, alkylene-aryl, -C(O)-alkyl, and -C(O)-aryl;

[0051] R 4 R 5 and R 5’ Each is an alkyl group;

[0052] R 6 and R 11 Each is independently -H or alkyl;

[0053] R 7 For fatty acid chains, -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 -γ-triazolyl-L;

[0054] Y is an alkylene group;

[0055] L is selected from fatty acid chains, -alkylene-C(O)-W, -alkylene-OC(O)-W, -alkylene-N-(alkylene-C(O)-NR) 11 -alkylene-NR 11The group consisting of -C(O)-W)2 and -alkylene-N-(alkylene-C(O)-W)2;

[0056] W represents a fatty acid chain, -O-alkylene-C(H)(OR) 8 )-alkylene-OR 9 Phospholipids or sterols;

[0057] R 8 and R 9 Each independently as R X or -C(O)-R X ;

[0058] R 10 R 22 R 33 R 33’ R 44 R 44’ R 55 and R 55’ Each independently is H or R A ;

[0059] R X It consists of fatty acid chains;

[0060] Each of the aforementioned alkyl, alkylene, alkylene-aryl, aryl, and triazolyl groups is optionally surrounded by one or more R groups. A Replace, where R A Each time it appears, it is independently selected from the following groups: hydrogen, halogen, 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 (where X is 0, 1, or 2), aryl, heteroaryl, heteroarylalkyl, cycloalkyl, heterocycloalkyl, and R B ;

[0061] R C and R D Each occurrence is independently selected from hydrogen, alkyl, haloalkyl, -C(O)R Band -C(O)OR B The group formed; or R C and R D Together with the nitrogen they are attached to, they form optional R A Substituted heterocycles; and R B It can be an alkyl, alkenyl, or alkynyl group optionally substituted with one or more fluorine groups.

[0062] In its implementation, this disclosure provides a method for treating cell proliferation disorders or sepsis in patients in need, comprising administering to the patient a therapeutically effective amount of the compounds or nanobiocompositions disclosed herein. Attached Figure Description

[0063] Figure 1 Cryo-TEM images of HDL-derived nanoparticles from entries A through F in Table 8 of Example 24 are shown. A 50 nm scale bar applies to all six images.

[0064] Figure 2 The Z-mean diameter and PDI value of nanoparticles in formulations A to F of formulation 1 in Example 23 were determined by DLS.

[0065] Figure 3 A graph showing the OD values ​​as a function of the test sample concentration in the NOD2 activation test described in Example 25.

[0066] Figure 4 The AE plotted the tumor growth curves of the study described in Example 26.

[0067] Figure 5 The AC plots show the tumor growth curves from the study described in Example 27.

[0068] definition

[0069] For convenience, certain terms used in the specification, embodiments, and claims are collected herein. 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 pertains.

[0070] Throughout this disclosure, various patents, patent applications, and publications are cited. The disclosures of these patents, patent applications, and publications, in their entirety, are incorporated herein by reference to provide a more complete description of the prior art known to those skilled in the art as of the date of this disclosure. In the event of any inconsistency between the cited patents, patent applications, and publications and this disclosure, this disclosure shall prevail.

[0071] The term "about," when immediately preceding a numerical value, indicates a range of additions or subtractions of variability acceptable in the art. In embodiments, the term "about" covers 10% of the value; for example, "about 50" means 45 to 55, "about 25,000" means 22,500 to 27,500, etc., unless the context of this disclosure otherwise or is inconsistent with such interpretation. For example, in a list of values ​​such as "about 49, about 50, about 55, ...", "about 50" means a range extending to less than half the interval between the preceding and subsequent values, for example, more than 49.5 to less than 52.5. Furthermore, the phrases "less than about" or "greater than about" for a value should be understood in light of the definition of the term "about" provided herein.

[0072] As used herein, “pharmaceutically acceptable” means compounds, materials, compositions, and / or dosage forms that, to a reasonable extent of medical judgment, are suitable for use in contact with human and animal tissues without excessive toxicity, irritation, allergic reactions, or other problems or complications, in proportion to a reasonable benefit / risk ratio.

[0073] "Salt" includes those obtained by reacting a compound acting as a base with an inorganic or organic acid to form a salt, or by reacting a compound acting as an acid with an inorganic or organic base to form a salt. "Salt" includes derivatives of active agents, wherein the active agent is modified by preparing its acid addition salt or base addition salt. 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 acids and organic acids. Representative examples of suitable inorganic acids include hydrochloric acid, hydrobromic acid, hydroiodic acid, phosphoric acid, sulfuric acid, and nitric acid. 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, 2-hydroxyethanesulfonic acid, tartaric acid, ascorbic acid, dihydroxynaphthyl acid, dimethylene salicylic acid, ethanedisulfonic acid, gluconic acid, citralic 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, hydroxynaphthyl carbamates, glycerophosphates, and ketoglutarate, etc. Base addition salts include, but are not limited to, ethylenediamine, N-methylglucosamine, lysine, arginine, ornithine, choline, N,N'-dibenzylethylenediamine, chloroprocaine, diethanolamine, procaine, N-benzylphenylethylamine, diethylamine, piperazine, tri-(hydroxymethyl)-aminomethane, tetramethylammonium hydroxide, triethylamine, dibenzylamine, ephenamine, dehydroabimethamine, N-ethylpiperidine, benzylamine, tetramethylammonium, tetraethylammonium, methylamine, dimethylamine, trimethylamine, ethylamine, basic amino acids such as lysine and arginine, and dicyclohexylamine. Examples of metal salts include lithium salts, sodium salts, potassium salts, magnesium salts, and calcium salts. Examples of ammonium salts and alkylated ammonium salts include ammonium salts, methylammonium salts, dimethylammonium salts, trimethylammonium salts, ethylammonium salts, hydroxyethylammonium salts, diethylammonium salts, butylammonium salts, and tetramethylammonium salts. Examples of organic bases include lysine, arginine, guanidine, diethanolamine, and choline. Standard methods for preparing pharmaceutically acceptable salts and their formulations are well known in the art and disclosed in various references, including, for example, "Remington: The Science and Practice of Pharmacy," A. Gennaro, ed., 20th edition, Lippincott, Williams & Wilkins, Philadelphia, PA.

[0074] As used interchangeably herein, the terms "carrier" or "vehicle" encompass carriers, excipients, adjuvants, and diluents, or any combination thereof, and refer to materials, compositions, or solvents involved in carrying or transporting a drug from one organ or part of the body to another organ or part of the body, such as liquid or solid fillers, diluents, excipients, solvents, or encapsulation materials. In addition to adjuvants, excipients, and diluents known to those skilled in the art, carriers also include nanoparticles of organic and inorganic properties.

[0075] For example, in embodiments, this disclosure provides nanoparticle carriers (e.g., HDL-derived nanoparticles) as delivery solvents for active agents (e.g., formulas (I), (IA), (IB), (II), (II-1), (II-2), (IIA), (IIA-1), (IIA-2), or compounds of Table 1). In embodiments, the drug is encapsulated within the nanoparticle carrier. In other embodiments, the drug is bound to the surface of the nanoparticle carrier. Association between the drug and the nanoparticle carrier can be achieved by a variety of means, including non-covalent binding and trapping the drug within the delivery solvent. In embodiments, the association is sufficiently stable such that the drug remains associated with the delivery solvent until it is delivered to the target site of the treated subject.

[0076] As used herein, the terms "drug combination," "therapeutic combination," or "combination" refer to a single dosage form comprising at least two therapeutically active agents, or separate dosage forms comprising at least two therapeutically active agents used together or separately for combination therapy. Administration of combination therapy includes sequential, simultaneous, or sequential administration of the same or different compositions and / or combinations via the same or different routes. For example, one therapeutically active agent may be formulated into one dosage form, and another therapeutically active agent may be formulated into a single or different dosage form. For example, one therapeutically active agent may be formulated into a solid oral dosage form, while a second therapeutically active agent may be formulated into a solution dosage form for parenteral administration. In embodiments, combination therapy optionally includes one or more pharmaceutically acceptable carriers or excipients, non-pharmaceuticalally active compounds, and / or inert substances.

[0077] As used herein, the phrase “a disease characterized by cell proliferation” or “a condition characterized by cell proliferation” includes, but is not limited to, cancer, benign tumors, and malignant tumors. Examples of cancer and tumors include, but are not limited to, cancer or tumor growths of the bladder, blood vessels, bones, brain, breast, cervix, chest, colon, endometrium, esophagus, eye, head, kidney, liver, lymph nodes, lung, oral cavity, neck, ovary, pancreas, prostate, rectum, colorectal, skin, stomach, testes, pharynx, thyroid, urothelial, and uterus.

[0078] The terms “treat,” “treating,” or “treatment” in connection with a specific disease or condition include prevention and / or reduction, improvement, amelioration, or elimination of the symptoms and / or pathology of a disease or condition. Generally, as used herein, the terms refer to the improvement, relief, reduction, and elimination of the symptoms of a disease or condition. The candidate compounds described herein may be present in a therapeutically effective amount in a formulation or pharmaceutical preparation, which is an amount that can cause biological effects such as apoptosis of certain cells (e.g., cancer cells), a reduction in the proliferation of certain cells, or an improvement, relief, reduction, or elimination of the symptoms of a disease or condition such as sepsis. The term may also refer to reducing or stopping the rate of cell proliferation (e.g., slowing or stopping tumor growth) or reducing the number of proliferating cancer cells (e.g., removing part or all of a tumor).

[0079] As used herein, the terms "patient" or "subject" include all mammals, and more specifically, humans. The methods described herein can be used for both human treatment and veterinary applications. In one implementation, the subject is a human.

[0080] As used herein, “prevention” or “preventing” means reducing the risk of developing a given disease or condition. For example, preventing at least one clinical symptom of the disease from occurring in subjects who may be exposed to or susceptible to the disease but have not yet experienced or exhibited symptoms of the disease.

[0081] As used herein, "therapeutic effective amount" means the amount of a compound or therapeutically active agent that, when administered to a subject for the treatment of a disease or other undesirable medical condition, is sufficient to have a beneficial effect on that disease or condition. Therapeutic effective amounts will vary depending on the type of compound or therapeutically active agent chosen, the disease or condition and its severity, and the age, weight, etc., of the patient to be treated.

[0082] The use of "optional" or "optionally" implies that the event or situation described below may or may not occur, and the description includes both the possibility that the event or situation occurs and the possibility that it does not occur. For example, "optionally substituted aryl" encompasses both "aryl" and "substituted aryl" as defined below. Those skilled in the art will understand that for any group containing one or more substituents, such groups are not intended to introduce any substitution or substituent form that is spatially impractical, synthetically infeasible, and / or inherently unstable.

[0083] It should also be noted that the claims can be drafted to exclude any optional elements. Therefore, this statement is intended to serve as an antecedent basis for the use of exclusive terms such as "unique" and "only" or for the use of a "negative" limitation in relation to the description of the claim elements.

[0084] When listing a range of values, the aim is to encompass all values ​​and subranges within that range. For example, "C1-C6 alkyl" is intended to cover C1, C2, C3, C4, C5, C6 ... 1-6 C 1-5 C 1-4 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.

[0085] As used herein, the term "acyl" refers to an RC(O)- group, such as, but not limited to, (alkyl)-C(O)-, (alkenyl)-C(O)-, (alkynyl)-C(O)-, (aryl)-C(O)-, (cycloalkyl)-C(O)-, (heteroaryl)-C(O)-, and (heterocyclic)-C(O)-, wherein the group is linked to the parent molecule structure via a carbonyl functional group. In an embodiment, it is C 1-10 An acyl group is defined as the carbonyl carbon atom of a chain or ring moiety, such as alkyl, alkenyl, alkynyl, aryl, cycloalkyl, or heteroaryl. For example, a C4 acyl group has three other ring or chain atoms plus a carbonyl group.

[0086] "alkyl" or "alkyl group" refers to a fully saturated straight-chain or branched hydrocarbon chain. In embodiments, the alkyl group comprises 1 to 30 carbon atoms. In embodiments, the alkyl group has 1 to 12 carbon atoms and is connected to the remainder of the molecule by a single bond. For example, alkyl groups comprising any number of carbon atoms from 1 to 12 are included. Alkyl groups comprising up to 12 carbon atoms are C1-C1. 12 Alkyl groups, alkyl groups containing up to 10 carbon atoms, are C1-C6. 10 Alkyl groups, specifically C1-C6 alkyl groups containing up to 6 carbon atoms and C1-C5 alkyl groups containing up to 5 carbon atoms, are further defined as alkyl groups. C1-C5 alkyl groups include C5 alkyl, C4 alkyl, C3 alkyl, C2 alkyl, and C1 alkyl (i.e., methyl). C1-C6 alkyl groups include all the portions described above for C1-C5 alkyl groups but also include C6 alkyl groups. C1-C10 Alkyl groups include all the portions described above for C1-C5 and C1-C6 alkyl groups, but also include C7, C8, C9, and C6 alkyl groups. 10 Alkyl group. Similarly, C1-C 12 Alkyl groups include all the foregoing portions, but also include C. 11 and C 12 Alkyl group. C1-C 12 Non-limiting examples of alkyl groups include methyl, ethyl, n-propyl, isopropyl, sec-propyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, tert-pentyl, n-hexyl, n-heptyl, n-octyl, n-nonyl, n-decyl, n-undecyl, and n-dodecyl. Unless otherwise specifically stated in the specification, the alkyl group may optionally be substituted. In embodiments, "alkyl" is a straight-chain hydrocarbon. In embodiments, "alkyl" is a branched-chain hydrocarbon.

[0087] "alkylene" or "alkylene chain" refers to a fully saturated straight-chain or branched divalent hydrocarbon chain. In the embodiments, the alkylene group has 1 to 12 carbon atoms. C1-C 12 Non-limiting examples of alkylene groups include methylene, ethylene, propylene, n-butylene, vinylene, propenylene, and n-butenylene. The alkylene chain is attached to the remainder of the molecule and to the functional group via single bonds. The connection points between the alkylene chain and the remainder of the molecule and to the functional group can be one carbon or any two carbons within the chain. Unless otherwise specified in the specification, the alkylene chain may optionally be substituted.

[0088] "Alkenyl" or "alkenyl group" refers to a straight-chain or branched hydrocarbon chain. In embodiments, the alkenyl group comprises 1 to 30 carbon atoms. In embodiments, the alkenyl group comprises 2 to 12 carbon atoms and has one or more carbon-carbon double bonds, such as a straight-chain or branched group with 2 to 8 carbon atoms, referred to herein as C 2- C8 alkenyl groups. Each alkenyl group is connected to the rest of the molecule via a single bond. This includes alkenyl groups containing any number of carbon atoms from 2 to 12. Alkenyl groups containing up to 12 carbon atoms are C2-C. 12 Alkenyl groups, specifically those containing up to 10 carbon atoms, are C2-C. 10 Alkenyl groups, specifically C2-C6 alkenyl groups containing up to six carbon atoms and C2-C5 alkenyl groups containing up to five carbon atoms, are further defined as alkenyl groups. C2-C5 alkenyl groups include C5, C4, C3, and C2 alkenyl groups. C2-C6 alkenyl groups include all the portions described above for C2-C5 alkenyl groups, but also include C6 alkenyl groups. C2-C 10 The alkenyl group includes all the portions described above for C2-C5 and C2-C6 alkenyl groups, but also includes C7, C8, C9, and C6 alkenyl groups. 10 Alkenyl. Similarly, C2-C12 The alkenyl group includes all the aforementioned parts, but also includes C. 11 and C 12 Alkenyl. C2-C 12 Non-limiting examples of alkenyl groups include ethenyl, 1-propenyl, 2-propenyl (allyl), isopropenyl, 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... Alkenyl, 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, 1-dodecenyl, and 11-dodecenyl. Unless otherwise specifically stated in the specification, the alkyl group may optionally be substituted.

[0089] "Alynyl" or "alkynyl group" refers to a straight-chain or branched hydrocarbon chain. In embodiments, the alkynyl group contains 1 to 30 carbon atoms. In embodiments, the alkynyl group contains 2 to 12 carbon atoms and has one or more carbon-carbon triple bonds, such as a straight-chain or branched group with 2 to 8 carbon atoms, referred to herein as C2-C8 alkynyl. Each alkynyl group is connected to the rest of the molecule by a single bond. This includes alkynyl groups containing any number of carbon atoms from 2 to 12. Alynyl groups containing up to 12 carbon atoms are C2-C8 alkynyl groups. 12 The alkynyl group, which contains up to 10 carbon atoms, is C2-C. 10 The ynyl group, comprising up to 6 carbon atoms, is C2-C6 ynyl, and comprising up to 5 carbon atoms, is C2-C5 ynyl. C2-C5 ynyl groups include C5, C4, C3, and C2 ynyl groups. C2-C6 ynyl groups include all the portions described above for C2-C5 ynyl groups, but also include C6 ynyl groups. C2-C 10The alkynyl group includes all the portions described above for C2-C5 and C2-C6 alkynyl groups, but also includes C7, C8, C9, and C6 alkynyl groups. 10 Alkyne group. Similarly, C2-C 12 The alkynyl group includes all the aforementioned parts, but also includes C. 11 and C 12 Alkyne group. C2-C 12 Non-limiting examples of alkenyl groups include ethynyl, propynyl, butynyl, and pentyynyl. Unless otherwise specified in the specification, alkyl groups may optionally be substituted.

[0090] "Aryl" refers to a hydrocarbon ring system comprising hydrogen, 6 to 18 carbon atoms, and at least one aromatic ring, which is linked to the remaining molecules by a single bond. For the purposes of this invention, the aryl group can be a monocyclic, bicyclic, tricyclic, tetracyclic, or other polycyclic system, which may include fused or bridged ring systems. Aryl groups include, but are not limited to, those derived from: aceanthrylene, acenaphthene, acephenanthrylene, anthracene, azurite, benzene, etc. Fluoranthene, fluorene, as-benzodiindene, s-benzodiindene, indene, indene, naphthalene, phenalene, pleiadene, pyrene, and benzo[a]phenanthrene. Unless otherwise specified in the specification, the aryl group may be optionally substituted.

[0091] "Arylalkyl" or "arylalkyl" refers to the formula -R b -R c The group, wherein R b For alkylene groups as defined above and R c It can be one or more aryl groups as defined above, such as benzyl and diphenylmethyl. Unless otherwise specified in the specification, the aryl group may optionally be substituted.

[0092] "Carbocyclic group," "carbocyclic ring," or "carbocycle" refers to a ring structure in which each of the ring-forming atoms is a carbon atom, and that the ring is connected to the rest of the molecule by a single bond. A carbocyclic ring can contain 3 to 20 carbon atoms. Carbocyclic rings include aryl groups as defined herein, as well as cycloalkyl, cycloalkenyl, and cycloynyl groups. Unless otherwise specifically stated in the specification, the carbocyclic group may optionally be substituted.

[0093] "Cycloalkyl" refers to a stable, non-aromatic, fully saturated monocyclic or polycyclic hydrocarbon consisting only of carbon and hydrogen atoms. It may include fused, spirocyclic, or bridged ring systems having 3 to 20 carbon atoms connected to the remainder of the molecule by single bonds. Monocyclic cycloalkyl groups include, for example, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, and cyclooctyl. Polycyclic cycloalkyl groups include, for example, adamantyl, norbornyl, decahydronaphthyl, and 7,7-dimethyl-bicyclo[2.2.1]heptyl. Unless otherwise specified in the specification, the cycloalkyl group may optionally be substituted.

[0094] "Cycloalkenyl" refers to a stable, non-aromatic monocyclic or polycyclic hydrocarbon consisting only of carbon and hydrogen atoms, having one or more carbon-carbon double bonds, which may include fused, spirocyclic, or bridged ring systems, having 3 to 20 carbon atoms, preferably 3 to 10 carbon atoms, and connected to the rest of the molecule by single bonds. Monocyclic cycloalkenyl groups include, for example, cyclopentenyl, cyclohexenyl, cycloheptenyl, and cyclooctenyl. Polycyclic cycloalkenyl groups include, for example, bicyclic [2.2.1]hept-2-enyl. Unless otherwise specifically stated in the specification, the cycloalkenyl group may optionally be substituted.

[0095] "Cycloynyl" refers to a stable, non-aromatic monocyclic or polycyclic hydrocarbon consisting only of carbon and hydrogen atoms, having 3 to 20 carbon atoms and one or more carbon-carbon triple bonds. It may include fused, spirocyclic, or bridged ring systems and is connected to the rest of the molecule by single bonds. Monocyclic cycloynyl groups include, for example, cycloheptynyl and cyclooctyynyl. Unless otherwise specified in the specification, the cycloynyl group may optionally be substituted.

[0096] "Heterocyclic group," "heterocyclic ring," or "heterocycle" refers to a stable 3- to 20-membered aromatic or non-aromatic ring consisting of 2 to 12 carbon atoms and 1 to 6 heteroatoms selected from the group consisting of nitrogen, oxygen, and sulfur. The heterocycle can be aromatic (heteroaryl) or non-aromatic. Unless otherwise specified in the specification, the heterocyclic group can be a monocyclic, bicyclic, tricyclic, tetracyclic, or other polycyclic system, which may include fused, spirocyclic, or bridged ring systems; and the nitrogen, carbon, or sulfur atoms in the heterocyclic group may optionally be oxidized; the nitrogen atom may optionally be quaternized; and the heterocyclic group may be partially or fully saturated. Examples of such heterocyclic groups include, but are not limited to, dioxanepentyl, thienyl[1,3]dithiaalkyl, decahydroisoquinolinyl, biotinyl, dihydrofuranyl, dihydroindolyl, dihydropyranyl, dihydrothienyl, dithiazolyl, homopiperidinyl, pyranyl, pyrazolinyl, thiaranyl, imidazolinyl, imidazoalkyl, isothiazolinyl, isoxazolinyl, morpholinyl, octahydroindolyl, octahydroisoindolyl, 2-oxopiperazinyl, 2 -Opiperidinyl, 2-oxopyrrolidinyl, oxazolyl, piperidinyl, piperazinyl, 4-piperidinyl, pyrrolidinyl, pyrrolidine-2-keto, pyrazolyl, quininecycloyl, thiazolyl, tetrahydrofuranyl, tetrahydroisoquinolinyl, trithiaalkyl, tetrahydropyranyl, thiomorpholinyl, thiamorpholinyl, 1-oxo-thiomorpholinyl, and 1,1-dioxo-thiomorpholinyl. Unless otherwise specified in the specification, the heterocyclic group may optionally be substituted.

[0097] "Heteroaryl" refers to a 5- to 20-membered ring system comprising a hydrogen atom, 1 to 13 carbon atoms, 1 to 6 heteroatoms selected from the group consisting of nitrogen, oxygen, and sulfur, and at least one aromatic ring. For the purposes of this disclosure, a heteroaryl can be a monocyclic, bicyclic, tricyclic, or tetracyclic system, which may include fused or bridged ring systems; a heteroaryl may comprise one or more non-aromatic rings (e.g., cycloalkyl or heterocyclic groups) fused to an aromatic ring. The nitrogen, carbon, or sulfur atom in the heteroaryl may optionally be oxidized; the nitrogen atom may optionally be quaternized. Examples include, but are not limited to, azepinyl, acridine, benzimidazolyl, benzothiazolyl, benzoindolyl, benzodioxolyl, benzofuranyl, benzooxazolyl, benzothiazolyl, benzothiadiazolyl, benzo[b][1,4]dioxepinyl, 1,4-benzodioxylalkyl, benzonaphthofuranyl, benzooxazolyl, benzodioxolyl, benzodioxolyl, benzodioxinyl, benzopyranyl, benzopyranonyl, benzofuranyl, benzofuranonyl, benzothienyl (benzothiophenyl) yl), benzotriazole, benzo[4,6]imidazo[1,2-a]pyridyl, carbazole, cenolinyl, dibenzofuranyl, dibenzothiophene, furanyl, furanone, isothiazolyl, imidazoyl, indazole, indolyl, indazole, isoindolyl, indololinyl, isoindololinyl, isoquinolinyl, indololinazinyl, isoxazolyl, naphridyl, oxadiazolyl, 2-oxoachenginyl, oxazolyl, ethylene oxide 1-Pyridyl oxide, 1-pyrimidinyl oxide, 1-pyrazinyl oxide, 1-pyridazinyl oxide, 1-phenyl-1H-pyrroloyl, phenazinyl, phenothiazinyl, phenotoxazinyl, phthalazinyl, pteridinyl, purine, pyrroloyl, pyrazolyl, pyridinyl, pyridinyl, pyrazinyl, pyrimidinyl, pyridazinyl, quinazolinyl, quinoxalinyl, quinolinyl, quininecycloyl, isoquinolinyl, tetrahydroquinolinyl, thiazolyl, thiadiazolyl, triazolyl, tetrazolyl, triazinyl, and phenylthio (i.e., thiophene). Unless otherwise specified in the specification, heteroaryl groups may optionally be substituted.

[0098] "Heteroarylalkyl" refers to the formula -R b -R f The group, wherein R b For alkylene chains as defined above and R f The heteroaryl group is as defined above. Unless otherwise specified in the specification, the heteroaryl alkyl group may optionally be substituted.

[0099] As used herein, the term "substituted" means any of the above groups (i.e., alkyl, alkenyl, alkynyl, aryl, arylalkyl, carbocyclic, cycloalkyl, cycloalkenyl, cycloalkynyl, heterocyclic, N-heterocyclic, heteroaryl, etc.) in which at least one hydrogen atom is replaced by a bond with, for example, but not limited to, the following non-hydrogen atoms: halogen atoms, such as F, Cl, Br, and I; oxygen atoms in groups such as hydroxyl, alkoxy, and ester; sulfur atoms in groups such as thioalkyl, thioalkyl, sulfonyl, sulfonyl, and sulfoxide; nitrogen atoms in groups such as amines, amides, alkylamines, dialkylamines, arylamines, alkylarylamines, diarylamines, N-oxides, imides, and enamines; silicon atoms in groups such as trialkylsilyl groups, dialkylarylsilyl groups, alkyldiarylsilyl groups, and triarylsilyl groups; and other heteroatoms in various other groups. "Substituted" also means any of the above groups in which one or more hydrogen atoms are replaced by a higher-order bond (e.g., a double or triple bond) with a heteroatom such as oxygen in oxo, carbonyl, carboxyl, and ester groups, or nitrogen in groups such as imine, oxime, hydrazone, and nitrile groups. For example, "substituted" includes any of the above groups in which one or more hydrogen atoms are replaced by NR g R h NR g C(=O)R h NR g C(=O)NR g R h NR g C(=O)OR h NR g SO2R h OC (=O)NR g R h OR g SR g SOR g SO2R g OSO2R g SO2OR g =NSO2R g and SO2NR g R h Substitution. "Substituted" also refers to any of the above groups, where one or more hydrogen atoms are replaced by C(=O)R. g C(=O)OR g C(=O)NR g R h CH2SO2R g CH2SO2NR g R h Instead. In the foregoing, Rg and R h The same or different and independently being hydrogen, alkyl, alkenyl, alkynyl, alkoxy, alkylamino, thioalkyl, aryl, aralkyl, cycloalkyl, cycloalkenyl, cycloalkynyl, cycloalkylalkyl, haloalkyl, haloalkenyl, haloalkynyl, heterocyclic, N-heterocyclic, heterocyclic alkyl, heteroaryl, N-heteroaryl and / or heteroarylalkyl. "Substituted" further means any of the above groups, wherein one or more hydrogen atoms are replaced by a bond with the following groups: amino, cyano, hydroxyl, imino, nitro, oxo, thioxo, halogen, alkyl, alkenyl, alkynyl, alkylamino, thioalkyl, aryl, aralkyl, cycloalkyl, cycloalkenyl, cycloalkynyl, cycloalkylalkyl, haloalkyl, haloalkenyl, haloalkynyl, heterocyclic, N-heterocyclic, heterocyclic alkyl, heteroaryl, N-heteroaryl and / or heteroarylalkyl. Furthermore, "substituted" means any of the above groups in which two hydrogen atoms are each free-bonded to form a fused ring system comprising the atom to which the hydrogen is bonded. Additionally, each of the aforementioned substituents may optionally be substituted by one or more of the above substituents.

[0100] The compounds disclosed herein may contain one or more chiral centers and / or double bonds, and thus exist as stereoisomers, such as geometric isomers, enantiomers, or diastereomers. The term "stereoisomer," as used herein, comprises all geometric isomers, enantiomers, or diastereomers. These compounds may be designated by the symbol "R" or "S," depending on the configuration of the substituents surrounding the stereocarbon atom. This disclosure covers various stereoisomers of these compounds and mixtures thereof. Stereoisomers include enantiomers and diastereomers. Mixtures of enantiomers or diastereomers may be designated as "(±)" in nomenclature, but those skilled in the art will recognize that the structure may implicitly represent the chiral center. In embodiments, enantiomers or stereoisomers may be provided without substantially containing the corresponding enantiomer.

[0101] In one embodiment, the compound is a racemic mixture of (S)-isomers and (R)-isomers. In other embodiments, a mixture of compounds is provided herein, wherein the individual compounds in the mixture are present primarily in either the (S)-isomer configuration or the (R)-isomer configuration. For example, the (S)-enantiomer excess of the compound mixture is 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 greater. In other embodiments, the (S)-enantiomer excess of the compound mixture is greater than about 55% to about 99.5%, greater than about 60% to about 99.5%, greater than about 65% to about 99.5%, greater than about 70% to about 99.5%, greater than about 75% to about 99.5%, greater than about 80% to about 99.5%, greater than about 85% to about 99.5%, greater than about 90% to about 99.5%, greater than about 95% to about 99.5%, greater than about 96% to about 99.5%, greater than about 97% to about 99.5%, greater than about 98% to greater than about 99.5%, greater than about 99% to about 99.5%, or greater. In other embodiments, the (R)-enantiomer purity of the compound mixture is 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 greater. In some other embodiments, the (R)-enantiomer excess of the compound mixture is greater than about 55% to about 99.5%, greater than about 60% to about 99.5%, greater than about 65% to about 99.5%, greater than about 70% to about 99.5%, greater than about 75% to about 99.5%, greater than about 80% to about 99.5%, greater than about 85% to about 99.5%, greater than about 90% to about 99.5%, greater than about 95% to about 99.5%, greater than about 96% to about 99.5%, greater than about 97% to about 99.5%, greater than about 98% to greater than about 99.5%, greater than about 99% to about 99.5%, or greater.

[0102] The various stereoisomers of the compounds disclosed herein can be prepared by synthesis from commercially available starting materials containing an asymmetric center or a stereocenter, or by preparing a racemic mixture and then by a resolution method known to those skilled in the art. Examples of such resolution methods include: (1) linking a mixture of enantiomers to a chiral auxiliary agent, separating the resulting mixture of diastereomers by recrystallization or chromatography, and releasing the optically pure product from the auxiliary agent; (2) salt formation using an optically active resolving agent; or (3) direct separation of the mixture of optically enantiomers on a chiral chromatographic column. The stereoisomer mixture can also be resolved into its component stereoisomers by known methods such as chiral phase gas chromatography, chiral phase high-performance liquid chromatography, crystallizing the compound into a chiral salt complex, or crystallizing the compound in a chiral solvent. The stereoisomers can also be obtained from stereoisomerically pure intermediates, reagents, and catalysts by known asymmetric synthetic methods.

[0103] Geometric isomers may also be present in the compounds disclosed herein. This disclosure covers a variety of geometric isomers and mixtures thereof resulting from arrangements of substituents around the carbon-carbon double bond or around the carbocyclic ring. Substituents around the carbon-carbon double bond are designated as being in the “Z” or “E” configuration, wherein the terms “Z” and “E” are used according to IUPAC standards. Unless otherwise stated, the description of the double bond encompasses both the E and Z isomers.

[0104] Substituents around a carbon-carbon double bond may optionally be referred to as "cis" or "trans," where "cis" indicates a substituent on the same side of the double bond, and "trans" indicates a substituent on the opposite side of the double bond. The arrangement of substituents around the carbon ring is designated as "cis" or "trans." The term "cis" indicates a substituent on the same side of the ring plane, and the term "trans" indicates a substituent on the opposite side of the ring plane. A mixture of compounds in which substituents are located on both the same and opposite sides of the ring plane is designated as "cis / trans."

[0105] The compounds disclosed herein may exist as tautomers and both tautomer forms are intended to be covered by the scope of this disclosure, even if only one tautomer structure is described.

[0106] As used herein, the term "isotope variant" is intended to include compounds that differ only in the presence of atoms enriched with one or more isotopes. Such compounds can be used as, for example, analytical tools, probes in biological experiments, or therapeutic agents. For instance, an "isotope variant" of a compound may contain one or more non-radioactive isotopes, such as, for example, deuterium (…). 2 H or D), carbon-13 (H or D), carbon-13 13 C) or nitrogen-15 ( 15N), etc. It will be understood that in compounds undergoing such isotopic substitution, the following atoms (in their presence) can be varied so that, for example, any hydrogen atom can be [amount missing]. 2 H / D, any carbon can be 13 C, or any nitrogen, can be 15 N, and the presence and location of such atoms can be determined within the scope of the art. Similarly, the invention may include the preparation of isotopic variants using radioisotopes, for example, in cases where the resulting compound can be used for pharmaceutical and / or substrate tissue distribution studies. Radioisotope tritium (i.e. 3 H) and carbon-14 (i.e. 14 C) They are particularly useful for this purpose due to their ease of introduction and convenient use as detection methods. Furthermore, it is possible to prepare, for example... 11 C 18 F, 15 O and 13 Compounds substituted with positron-emitting isotopes such as N, and said compounds would be useful in positron emission tomography (PET) studies for examining substrate receptor occupancy.

[0107] As used herein, the term "triglyceride" refers to an ester derived from glycerol and three fatty acids. The fatty acids may be the same or different. The notation used to describe triglycerides in this specification is the same as that used to describe fatty acids below. Fatty acids may be attached to the glycerol molecule in any order; for example, any fatty acid may react with any hydroxyl group of the glycerol molecule to form an ester bond. For example, in non-limiting instances, triglycerides may comprise any combination of glycerol with the following fatty acids: C18:1, C14:1, C16:1, polyunsaturated, and saturated. A C18:1 fatty acid triglyceride 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 and three fatty acids, each with 18 carbon atoms, wherein each fatty acid has a double bond. Similarly, a C14:1 triglyceride is an ester of glycerol and three fatty acids, each with 14 carbon atoms, wherein each fatty acid has a double bond. Similarly, a C16:1 triglyceride is an ester of glycerol and three fatty acids, each with 16 carbon atoms, wherein each fatty acid has a double bond. A triglyceride of a C18:1 fatty acid in combination with a C14:1 and / or C16:1 fatty acid means: (a) a C18:1 triglyceride combined with a C14:1 triglyceride or a C16:1 triglyceride or a mixture of 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, while the other two are derived from or based on a C14:1 fatty acid and / or a C16:1 fatty acid.

[0108] The term "fatty acid" and similar terms refer to a carboxylic acid having a long aliphatic tail, whether 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 to C30 (e.g., C6 to C30), saturated or unsaturated, cis or trans, unsubstituted or substituted, branched or unbranched hydrocarbon chains (e.g., fatty acid chain lengths include C4 to C30 (e.g., C6 to 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, 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 saturated or unsaturated, cis or trans, unsubstituted or substituted hydrocarbon chains. 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 any double bonds. In embodiments, fatty acids are described herein by a capital letter "C" representing a carbon atom, followed by a number describing 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 represents a fatty acid with 16 carbon atoms having one double bond, such as palmitoleic acid. In this notation, the number after the colon does not specify the position of the double bond in the fatty acid, nor does it specify whether the hydrogen atoms bonded to the carbon atoms of the double bond are cis-bonded. 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).

[0109] The term "sterol" (e.g., but not limited to cholesterol) may also be used for the methods and compounds described herein. A sterol is an animal or plant steroid containing only a hydroxyl group at C-3 and no other functional groups. Typically, sterols contain 27 to 30 carbon atoms and a double bond at the 5 / 6 position, occasionally at the 7 / 8, 8 / 9, or other positions. Other sterols, besides these unsaturated forms, are saturated compounds obtainable by hydrogenation. A suitable example of an animal sterol is cholesterol. Typical examples of suitable plant sterols (preferred from an application standpoint) are ergosterol, campesterol, stigmasterol, brassosterol, and, preferably, sitosterol or dihydrositosterol, and more particularly, β-sitosterol or β-dihydrositosterol. In addition to the plant sterols mentioned, their esters are also preferred. The acid component of an ester can be traced back to a carboxylic acid corresponding to formula (CA-I): RICO-OH(CA-I); where RI CO is an aliphatic, linear, or branched acyl group containing 2 to 30 carbon atoms and O and / or 1, 2, or 3 double bonds. Typical examples include acetic acid, propionic acid, hexanoic acid, butyric acid, valeric acid, caproic acid, 2-ethylhexanoic acid, decanoic acid, cyclopentanepropionic acid, lauric acid, isotriadecanoic acid, myristic acid, palmitic acid, palmitoleic acid, stearic acid, isostearic acid, oleic acid, transoleic acid, petroselic acid, linoleic acid, conjugated linoleic acid (CLA), linolenic acid, tung oil acid, arachidic acid, codoleic acid, behenic acid, and erucic acid.

[0110] The term "phospholipid" refers to an amphiphilic compound consisting of two hydrophobic fatty acid "tails" and a hydrophilic "head" composed of a phosphate group. The two components are linked together by a glycerol molecule. The phosphate group can be modified with simple organic molecules such as choline, ethanolamine, or serine. Choline is an essential bioactive nutrient with the chemical formula R-(CH2)2-N(CH2)4. When the phosphate group is R-, it is called phosphocholine.

[0111] As used herein, "hemolyzed lipids" include (acyl-, single-chain) such as, in non-limiting embodiments, 1-myristoyl-2-hydroxy-sn-glycerol-3-phosphate choline (MHPC), 1-palmitoyl-2-hydroxy-sn-glycerol-3-phosphate choline (PHPC), and 1-stearoyl-2-hydroxy-sn-glycerol-3-phosphate choline (SHPC).

[0112] The terms "apolipoprotein AI" or "apoA-I" and "apolipoprotein Al" or "apoAl" refer to the protein encoded by the APOAI gene in humans. Detailed Implementation

[0113] The immune system in vertebrates is typically divided into two parts. The first part, the innate immune system, provides an initial response to infection within minutes to hours. Its cellular components include natural killer (NK) cells, innate lymphocytes (ILCs), and phagocytes such as monocytes, macrophages, and neutrophils. The innate immune system acts as a rapid first line of defense, triggered by the recognition of pathogens or endogenous danger signals via pattern recognition receptors (PRRs). Upon detection of pathogen-associated molecular patterns (PAMPs), the PRR initiates an innate immune response, which involves activating the subsequent adaptive immune system through antigen presentation, co-stimulation, and cytokine secretion. Furthermore, the PRR recognizes damage-associated molecular patterns (DAMPs), leading to a non-infectious inflammatory response. The second phase of the response to infection involves the second part of the immune system—the adaptive response—where T and B lymphocytes specifically recognize the pathogen, proliferate, and become active against it. These cells also establish an immune memory for that specific infection. The specificity of the adaptive immune system response is mediated by recombination of immunoglobulin genes at the lymphocyte level. Immune memory leads to a faster and more quantitative immune response to previously encountered antigens (compared to the initial response alone). Although the innate immune system has long been thought to lack memory, recent studies have shown that innate immune cells undergo metabolic and epigenetic reorganization, adjusting their functional programs in a process known as 'training immunity', which is believed to be innate immune memory.

[0114] Training immunity is defined by secondary, long-term hyperresponsiveness, characterized by increased cytokine secretion resulting from metabolic and epigenetic recombination following initial injury to myeloid cells and their progenitors, as well as stem cells in the bone marrow, spleen, and blood. Training immunity (also known as innate immune memory) is also defined by long-term enhanced responsiveness (e.g., high cytokine production) following secondary stimulation with bone marrow innate immune cells, induced by initial injury to these cells or their progenitors, and stem cells in the bone marrow and spleen, and mediated by epigenetic, metabolic, and transcriptional recombination.

[0115] Training immunity is modulated and maintained by inducing training properties in progenitor cells in the bone marrow, resulting in persistent reprogramming beyond the lifespan of myeloid cells in the bloodstream. While training immunity can be induced in cultured myeloid cells using a series of 'training agents', its systemic induction requires cell engagement of bone marrow progenitor cells.

[0116] In one aspect, this disclosure provides compounds (e.g., compounds of formula (I), (IA), (IB), (II), (II-1), (II-2), (IIA), (IIA-1), (IIA-2), or the compounds in Table 1) that activate protein 2 (NOD2) containing a nucleotide-binding oligomerization domain. This disclosure also provides nanobiocompositions comprising a nanoparticle carrier (e.g., HDL-derived nanoparticles) containing compounds of this disclosure (e.g., compounds of formula (I), (IA), (IB), (II), (II-1), (II-2), (IIA), (IIA-1), (IIA-2), or the compounds in Table 1). The nanobiocompositions of this disclosure comprising compounds of this disclosure that activate protein 2 (NOD2) containing a nucleotide-binding oligomerization domain are designed to exhibit proclivity. These nanomaterials can be administered (e.g., intravenously) to promote immune training. Therapeutic induction of immune training can be used, for example, to overcome immune paralysis in sepsis and infections, to treat cell proliferation disorders (e.g., cancer), and to enhance immune responses.

[0117] compound

[0118] In the embodiments, this disclosure provides compounds of formula (I) or pharmaceutically acceptable salts thereof:

[0119]

[0120] in:

[0121] R 1 -H or -C(O)-R X ;

[0122] R 2 and R 3 Each is independently selected from the group consisting of -H, alkyl, alkylene-aryl, -C(O)-alkyl, and -C(O)-aryl;

[0123] R 4 R 5 and R 5’ Each is an alkyl group;

[0124] R 6 and R 11 Each is independently -H or alkyl;

[0125] R 7 For fatty acid chains, -YN(R) 6 )-C(O)-O-alkylene-C(H)(OR8 )-alkylene-OR 9 -YN(R) 6 )-C(O)-R X -YOP(O)(OH)-O-alkylene-C(H)(OR) 8 )-alkylene-OR 9 Or -γ-triazolyl-L;

[0126] Y is an alkylene group;

[0127] L is selected from fatty acid chains, -alkylene-C(O)-W, -alkylene-OC(O)-W, -alkylene-N-(alkylene-C(O)-NR) 11 -alkylene-NR 11 The group consisting of -C(O)-W)2 and -alkylene-N-(alkylene-C(O)-W)2;

[0128] W represents a fatty acid chain, -O-alkylene-C(H)(OR) 8 )-alkylene-OR 9 Phospholipids or sterols;

[0129] R 8 and R 9 Each independently as R X or -C(O)-R X ;

[0130] R 22 R 33 R 33’ R 44 R 44’ R 55 and R 55’ Each independently is H or R A ;

[0131] R X It consists of fatty acid chains;

[0132] Each of the aforementioned alkyl, alkylene, alkylene-aryl, aryl, and triazolyl groups is optionally surrounded by one or more R groups. A Replace, where R A Each time it appears, it is independently selected from the following groups: hydrogen, halogen, 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 -NRC C(O)OR B -OC(O)R B -C(O)OR B -C(O)R B -CO2H, -NO2, -SH, S(O) X R B (where X is 0, 1, or 2), aryl, heteroaryl, heteroarylalkyl, cycloalkyl, heterocycloalkyl, and R B ;

[0133] R C and R D Each occurrence is independently selected from hydrogen, alkyl, haloalkyl, -C(O)R B and -C(O)OR B The group formed; or R C and R D Together with the nitrogen they are attached to, they form optional R A Substituted heterocycles; and

[0134] R B It can be an alkyl, alkenyl, or alkynyl group optionally substituted with one or more fluorine groups.

[0135] In the embodiments, this disclosure provides compounds of formula (I) or pharmaceutically acceptable salts thereof:

[0136]

[0137] in:

[0138] R 1 -H or -C(O)-R X ;

[0139] R 2 and R 3 Each is independently selected from the group consisting of -H, alkyl, alkylene-aryl, -C(O)-alkyl, and -C(O)-aryl;

[0140] R 4 R 5 and R 5’ Each is an alkyl group;

[0141] R 6 and R 11 Each is independently -H or alkyl;

[0142] R 7 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 -γ-triazolyl-L;

[0143] R Z and R Z’ Each independently is C 8-30 Fatty acid chain or -C(O)-C 16-30 fatty acid chains;

[0144] Y is an alkylene group;

[0145] L is selected from fatty acid chains, -alkylene-C(O)-W, -alkylene-OC(O)-W, -alkylene-N-(alkylene-C(O)-NR) 11 -alkylene-NR 11 The group consisting of -C(O)-W)2 and -alkylene-N-(alkylene-C(O)-W)2;

[0146] W represents a fatty acid chain, -O-alkylene-C(H)(OR) 8 )-alkylene-OR 9 Phospholipids or sterols;

[0147] R 8 and R 9 Each independently as R X or -C(O)-R X ;

[0148] R 10 R 22 R 33 R 33’ R 44 R 44’ R 55 and R 55’ Each independently is H or R A ;

[0149] R X It consists of fatty acid chains;

[0150] Each of the aforementioned alkyl, alkylene, alkylene-aryl, aryl, and triazolyl groups is optionally surrounded by one or more R groups. A Replace, where R AEach time it appears, it is independently selected from the following groups: hydrogen, halogen, 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 (where X is 0, 1, or 2), aryl, arylalkyl, heteroaryl, heteroarylalkyl, cycloalkyl, heterocycloalkyl, and R B ;

[0151] R C and R D Each occurrence is independently selected from hydrogen, alkyl, haloalkyl, -C(O)R B and -C(O)OR B The group formed; or R C and R D Together with the nitrogen they are attached to, they form optional R A Substituted heterocycles; and

[0152] R B The alkyl, alkenyl, or alkynyl groups may be optionally substituted with one or more fluorine groups;

[0153] Where R 7 C 9-30 When fatty acid chains are in the form of R 2 It is -H.

[0154] In embodiments of the compound of formula (I), the compound is a compound of formula (IA) or a pharmaceutically acceptable salt thereof:

[0155]

[0156] in:

[0157] R 1 -H or -C(O)-R X ;

[0158] R 2 and R 3Each is independently selected from the group consisting of -H, alkyl, alkylene-aryl, -C(O)-alkyl, and -C(O)-aryl;

[0159] R 4 R 5 and R 5’ Each is an alkyl group;

[0160] R 6 and R 11 Each is independently -H or alkyl;

[0161] R 7 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 -γ-triazolyl-L;

[0162] R Z and R Z’ Each independently is C 8-30 Fatty acid chain or -C(O)-C 16-30 fatty acid chains;

[0163] Y is an alkylene group;

[0164] L is selected from fatty acid chains, -alkylene-C(O)-W, -alkylene-OC(O)-W, -alkylene-N-(alkylene-C(O)-NR) 11 -alkylene-NR 11 The group consisting of -C(O)-W)2 and -alkylene-N-(alkylene-C(O)-W)2;

[0165] W represents a fatty acid chain, -O-alkylene-C(H)(OR) 8 )-alkylene-OR 9 Phospholipids or sterols;

[0166] R 8 and R 9 Each independently as R X or -C(O)-R X ;

[0167] R 10 R 22 R 33 R 33’ R 44 R 44’ R 55 and R 55’ Each independently is H or R A ;

[0168] R X It consists of fatty acid chains;

[0169] Each of the aforementioned alkyl, alkylene, alkylene-aryl, aryl, and triazolyl groups is optionally surrounded by one or more R groups. A Replace, where R A Each time it appears, it is independently selected from the following groups: hydrogen, halogen, 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 (where X is 0, 1, or 2), aryl, arylalkyl, heteroaryl, heteroarylalkyl, cycloalkyl, heterocycloalkyl, and R B ;

[0170] R C and R D Each occurrence is independently selected from hydrogen, alkyl, haloalkyl, -C(O)R B and -C(O)OR B The group formed; or R C and R D Together with the nitrogen they are attached to, they form optional R A Substituted heterocycles; and

[0171] R B The alkyl, alkenyl, or alkynyl groups may be optionally substituted with one or more fluorine groups;

[0172] Where R 7 C 9-30 When fatty acid chains are in the form of R2 It is -H.

[0173] In the implementation scheme, the compound of formula (I) is the compound of formula (IB) or a pharmaceutically acceptable salt thereof:

[0174]

[0175] In embodiments of compounds of formula (I), (IA), or (IB), Y is optionally -C(O)N(R) C (R) D ) substituted alkylene groups. In the embodiments, Y is -C 1-6 Alkylene. In the embodiments, Y is -CH2-. In the embodiments, Y is -CH2- or In the implementation scheme, Y is or In the implementation plan, Y is...

[0176] In embodiments of compounds of formula (I), (IA), or (IB). R 7 It is -γ-triazolyl-L.

[0177] In embodiments of compounds of formula (I), (IA), or (IB), R 7 It contains a cholesterol moiety or at least one fatty acid chain containing at least 17 carbons. In the embodiment, R 7 It contains a cholesterol-based moiety. In the implementation scheme, R... 7 It contains at least one fatty acid chain containing at least 17 carbons. In the implementation, R 7 It contains at least two fatty acid chains, each containing at least 17 carbons. In the implementation, R... 7 Contains at least two Cs 17 Fatty acid chain. In the implementation scheme, C 17 The fatty acid chain is derived from stearic acid or oleic acid. In the implementation scheme, C 17 The fatty acid chain is derived from stearic acid. In the implementation scheme, C 17 The fatty acid chain originates from oleic acid fatty acids. In the implementation scheme, R 7 Contains two Cs derived from stearic acid 17 Fatty acid chains.

[0178] In embodiments of compounds of formula (I), (IA), or (IB), R 7 It is an alkyl group having at least 16 carbons. In embodiments of compounds of formula (I), (IA), or (IB), R 7 It has at least 16 alkenyl groups. In embodiments of compounds of formula (I), (IA), or (IB), R 7It is an alkyl group having at least 18 carbons. In embodiments of the compound of formula (I), R 7 It is an alkenyl group having at least 18 carbons.

[0179] In embodiments of compounds of formula (I), (IA), or (IB), R 7 C 9-30 Fatty acid chain. In embodiments of the compound of formula (I), R 7 -C 9-30 Alkyl or C 9-30 Alkenyl. In the embodiment, R 7 -C 9-30 Alkyl or C 9-30 alkenyl, provided that R 7 -C 9-30 When alkyl, then R 2 For -H. In embodiments of the compound of formula (I), R 7 -C 9-30 Alkyl group. In the implementation scheme, R 7 -C 9-30 Alkyl and R 2 For -H. In embodiments of the compound of formula (I), R 7 -C 9-30 Alkenyl. In the embodiment, R 7 -C 15-30 Alkyl group. In the embodiment, R 7 -C 15-30 alkyl groups and R 2 For -H. In the implementation scheme, R 7 -C 15-30 Alkenyl group. In the embodiment, R 7 -C 17-19 Alkyl group. In the implementation scheme, R 7 -C 17-19 Alkyl and R 2 For -H. In the implementation scheme, R 7 -C 17-19 Alkenyl. In the embodiment, R 7 -C 18 Alkyl group. In the embodiment, R 7 -C 18 alkyl groups and R 2 For -H. In the implementation scheme, R 7 -C 18 Alkenyl group.

[0180] In embodiments of compounds of formula (I), (IA), or (IB), R 7 for:

[0181]

[0182] In embodiments of compounds of formula (I), (IA), or (IB), R 7 for:

[0183]

[0184] In embodiments of compounds of formula (I), (IA), or (IB), R 7 for:

[0185] And R 2 It is -H.

[0186] In embodiments of compounds of formula (I), (IA), or (IB), R 7 -YN(R) 11 )-C(O)-O-alkylene-C(H)(OR 8 )-alkylene-OR 9 .

[0187] In embodiments of compounds of formula (I), (IA), or (IB), R 7 -C(R) 10 (C(O)NH2)-alkylene-N(R) 11 )-C(O)-C 16-30 Fatty acid chain. In the implementation scheme, R 7 -C(H)(C(O)NH2)-C5 alkylene-N(R) 11 )-C(O)-C 17-30 Fatty acids. In the implementation plan, R 7 -C(H)(C(O)NH2)-alkylene-N(R) 11 )-C(O)-C 17-30 fatty acid.

[0188] In embodiments of compounds of formula (I), (IA), or (IB), R 7 -C(R) 10 (C(O)NH2)-alkylene-N(R) 11 )-C(O)-C 16-30 Alkyl group. In the implementation scheme, R 7 -C(R) 10 (C(O)NH2)-alkylene-N(R) 11 )-C(O)-C 17-30 Alkyl group. In the implementation scheme, R 7 -C(H)(C(O)NH2)-alkylene-N(R) 11)-C(O)-C 17-30 Alkyl group. In the implementation scheme, R 7 -C(H)(C(O)NH2)-C4 alkylene-N(R) 11 )-C(O)-C 17-30 Alkyl group. In embodiments of compounds of formula (I), R 7 -C(R) 10 (C(O)NH2)-alkylene-N(R) 11 )-C(O)-C 16-30 Alkyl and R 2 It is an alkylene-aryl group (e.g., benzyl). In the embodiments, R 7 -C(R) 10 (C(O)NH2)-alkylene-N(R) 11 )-C(O)-C 17-30 Alkyl and R 2 It is an alkylene-aryl group (e.g., benzyl). In the embodiments, R 7 -C(H)(C(O)NH2)-alkylene-N(R) 11 )-C(O)-C 17-30 Alkyl and R 2 It is an alkylene-aryl group (e.g., benzyl). In the embodiments, R 7 -C(H)(C(O)NH2)-C4 alkylene-N(R) 11 )-C(O)-C 17-30 Alkyl and R 2 It is an alkylene-aryl group (e.g., benzyl).

[0189] In embodiments of compounds of formula (I), (IA), or (IB), R 7 for

[0190] -(CR 10 R 10 )2-OP(O)(OH)-O-alkylene-C(R 10 (OR) Z )-alkylene-OR Z’ In embodiments of the compound of formula (I), R 7 is -CH2CH2-OP(O)(OH)-O-CH2-C(H)(OR Z )-CH2-OR Z’ In the implementation plan, R Z and R Z’ Each independently is C 12-20 Alkyl or -C(O)-C 16-30 Fatty acid chain. In the implementation scheme, R Z and RZ’ Each independently is C 18 Alkyl or -C(O)-C 17 Alkyl group. In the implementation scheme, R Z and R Z’ Each is independently -C(O)-C 16-30 Alkyl group. In the implementation scheme, R Z and R Z’ Both are -C(O)-C 17 alkyl.

[0191] In embodiments of compounds of formula (I), (IA), or (IB), R 7 -YN(R) 6 )-C(O)-O-alkylene-C(H)(OR 8 )-alkylene-OR 9 In the implementation plan, R 8 and R 9 Each independently is C 8-30 Alkyl or -C(O)-C 8-30 Alkyl group. In the implementation scheme, R 8 and R 9 Each independently is C 12-20 Alkyl or -C(O)-C 11-20 Alkyl group. In the implementation scheme, R 8 and R 9 Each independently is C 18 Alkyl or -C(O)-C 17 Alkyl group. In the implementation scheme, R 8 and R 9 Both are -C(O)-C 17 alkyl.

[0192] In embodiments of compounds of formula (I), (IA), or (IB), R A Each time it appears, it is independently selected from the following groups: hydrogen, halogen, 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 (where X is 0, 1 or 2), aryl, arylalkyl, heteroaryl, heteroarylalkyl, cycloalkyl, heterocycloalkyl.

[0193] In embodiments of compounds of formula (I), (IA), or (IB), R A Each time it appears, it is independently selected from the following groups: halogen, 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 (where X is 0, 1 or 2), aryl, arylalkyl, heteroaryl, heteroarylalkyl, cycloalkyl, heterocycloalkyl.

[0194] In the implementation scheme, the compound of formula (I) is the compound of formula (II) or a pharmaceutically acceptable salt thereof:

[0195]

[0196] in:

[0197] X1 is -N- and X2 is -C-; or X1 is -C- and X2 is -N-;

[0198] R 2 and R 3 Each is independently selected from the group consisting of -H, alkyl, aryl, alkylene-aryl, -C(O)-alkyl, and -C(O)-aryl;

[0199] R 4 R 5 and R 5’ Each is an alkyl group;

[0200] R 6 and R 11 Each is independently -H or alkyl;

[0201] Y is an alkylene group;

[0202] L is selected from fatty acid chains, -alkylene-C(O)-W, -alkylene-OC(O)-W, -alkylene-N-(alkylene-C(O)-NR) 11 -alkylene-NR 11 The group consisting of -C(O)-W)2 and -alkylene-N-(alkylene-C(O)-W)2;

[0203] W represents a fatty acid chain, -O-alkylene-C(H)(OR) 8 )-alkylene-OR 9 Phospholipids or sterols;

[0204] R 8 and R 9 Each independently as R X or -C(O)-R X ;

[0205] R X It consists of fatty acid chains;

[0206] Each of the aforementioned alkyl, alkylene, alkylene-aryl, and aryl groups is optionally surrounded by one or more R groups. A replace;

[0207] R A Each time it appears, it is independently selected from the following groups: hydrogen, halogen, 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 (where X is 0, 1, or 2), aryl, arylalkyl, heteroaryl, heteroarylalkyl, cycloalkyl, heterocycloalkyl, and R B ;

[0208] R C and R D Each occurrence is independently selected from hydrogen, alkyl, haloalkyl, -C(O)R B and -C(O)OR B The group formed; or R C and R DTogether with the nitrogen they are attached to, they form optional R A Substituted heterocycles; and

[0209] R B It can be an alkyl, alkenyl, or alkynyl group optionally substituted with one or more fluorine groups.

[0210] In the implementation scheme, the compound of formula (II) is:

[0211]

[0212] In the implementation scheme, the compound of formula (II) is a compound of formula (II-1) or a pharmaceutically acceptable salt thereof.

[0213]

[0214] In the implementation scheme, the compound of formula (II-1) is Or its pharmaceutically acceptable salt.

[0215] In the implementation scheme, the compound of formula (II) is the compound of formula (II-2):

[0216]

[0217] In the implementation scheme, the compound of formula (II-2) is:

[0218]

[0219] Or its pharmaceutically acceptable salt.

[0220] In the implementation scheme, the compound of formula (I) is a compound of formula (IIA) or a pharmaceutically acceptable salt thereof:

[0221]

[0222] in:

[0223] X1 is -N- and X2 is -C-; or X1 is -C- and X2 is -N-

[0224] R 2 and R 3 Each is independently selected from the group consisting of -H, alkyl, aryl, alkylene-aryl, -C(O)-alkyl, and -C(O)-aryl;

[0225] R 4 R 5 and R 5’ Each is an alkyl group;

[0226] R 6 and R 11 Each is independently -H or alkyl;

[0227] Y is an alkylene group;

[0228] L is selected from fatty acid chains, -alkylene-C(O)-W, -alkylene-OC(O)-W, -alkylene-N-(alkylene-C(O)-NR) 11 -alkylene-NR 11 The group consisting of -C(O)-W)2 and -alkylene-N-(alkylene-C(O)-W)2;

[0229] W represents a fatty acid chain, -O-alkylene-C(H)(OR) 8 )-alkylene-OR 9 Phospholipids or sterols;

[0230] R 8 and R 9 Each independently as R X or -C(O)-R X ;

[0231] R X It consists of fatty acid chains;

[0232] Each of the aforementioned alkyl, alkylene, alkylene-aryl, and aryl groups is optionally surrounded by one or more R groups. A replace;

[0233] R A Each time it appears, it is independently selected from the following groups: hydrogen, halogen, 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 (where X is 0, 1, or 2), aryl, arylalkyl, heteroaryl, heteroarylalkyl, cycloalkyl, heterocycloalkyl, and R B ;

[0234] R C and R D Each occurrence is independently selected from hydrogen, alkyl, haloalkyl, -C(O)R Band -C(O)OR B The group formed; or R C and R D Together with the nitrogen they are attached to, they form optional R A Substituted heterocycles; and

[0235] R B It can be an alkyl, alkenyl, or alkynyl group optionally substituted with one or more fluorine groups.

[0236] In the implementation scheme, the compound of formula (IIA) is:

[0237]

[0238] In the implementation scheme, the compound of formula (IIA) is a compound of formula (IIA-1) or a pharmaceutically acceptable salt thereof:

[0239]

[0240] In the implementation scheme, the compound of formula (IIA-1) is:

[0241]

[0242] Or its pharmaceutically acceptable salt.

[0243] In the implementation scheme, the compound of formula (IIA) is a compound of formula (IIA-2) or a pharmaceutically acceptable salt thereof:

[0244]

[0245] In the implementation scheme, the compound of formula (IIA-2) is:

[0246]

[0247] Or its pharmaceutically acceptable salt.

[0248] In embodiments of compounds of formula (I), (IA), (IB), (II), (II-1), (II-2), (II-A), (IIA-1), or (IIA-2), Y is alkylene. In embodiments of compounds of formula (I), (IA), (IB), (II), (II-1), (II-2), (II-A), (IIA-1), or (IIA-2), Y is C 1-6 Alkylene. In the embodiment, Y is C 1-5 Alkylene. In the embodiment, Y is C 1-3 Alkylene. In the embodiments, Y is optionally -C(O)N(R) C (R) D ) substituted alkylene groups, wherein RC and R D Defined herein. In the implementation, Y is -CH2-. In the implementation, Y is... In the implementation plan, Y is...

[0249] In embodiments of compounds of formula (I), (IA), (IB), (II), (II-1), (II-2), (II-A), (IIA-1), or (IIA-2), R A Each time it appears, it is independently selected from the following groups: hydrogen, halogen, 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 (where X is 0, 1, or 2), aryl, arylalkyl, heteroaryl, heteroarylalkyl, cycloalkyl, heterocycloalkyl, and R B .

[0250] In embodiments of compounds of formula (I), (IA), (IB), (II), (II-1), (II-2), (II-A), (IIA-1), or (IIA-2), R A Each time it appears, it is independently selected from the following groups: halogen, 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 (where X is 0, 1, or 2), aryl, arylalkyl, heteroaryl, heteroarylalkyl, cycloalkyl, heterocycloalkyl, and R B .

[0251] In embodiments of compounds of formula (I), (IA), (IB), (II), (II-1), (II-2), (II-A), (IIA-1), or (IIA-2), the alkyl, alkylene, alkylene-aryl, and aryl groups are optionally separated by one or more R groups. A replace;

[0252] In embodiments of compounds of formula (I), (IA), (IB), (II), (II-1), (II-2), (II-A), (IIA-1), or (IIA-2), R C and R D Each occurrence is independently selected from hydrogen, alkyl, haloalkyl, -C(O)R B and -C(O)OR B The group formed; or R C and R D Together with the nitrogen they are attached to, they form optional R A Substituted heterocycles.

[0253] In embodiments of compounds of formula (I), (IA), (IB), (II), (II-1), (II-2), (II-A), (IIA-1), or (IIA-2), R B It can be an alkyl, alkenyl, or alkynyl group optionally substituted with one or more fluorine groups.

[0254] In embodiments of compounds of formula (I), (IA), (IB), (II), (II-1), (II-2), (II-A), (IIA-1), or (IIA-2), R A It is -H.

[0255] In some embodiments of compounds of formula (I), (IA), (IB), (II), (II-1), (II-2), (II-A), (IIA-1), or (IIA-2), R 2 It is alkyl, aryl, or alkylene-aryl. In embodiments, the aryl group is optionally substituted with an alkyl group.

[0256] In embodiments of compounds of formula (I), (IA), (IB), (II), (II-1), (II-2), (II-A), (IIA-1), or (IIA-2), R 2 and R 3Each is independently selected from the group consisting of -H, alkyl, aryl, alkylene-aryl, -C(O)-alkyl, and -C(O)-aryl.

[0257] In embodiments of compounds of formula (I), (IA), (IB), (II), (II-1), (II-2), (II-A), (IIA-1), or (IIA-2), R 2 It is -H or benzyl. In embodiments of the compounds of formula (I), R 2 For -H. In the implementation scheme, R 2 It is benzyl.

[0258] In embodiments of compounds of formula (I), (IA), (IB), (II), (II-1), (II-2), (II-A), (IIA-1), or (IIA-2), R 4 R 5 and R 5’ Each is an alkyl group.

[0259] In embodiments of compounds of formula (I), (IA), (IB), (II), (II-1), (II-2), (II-A), (IIA-1), or (IIA-2), R 4 It is an alkyl group. In the embodiment, R... 4 It is a methyl group.

[0260] In embodiments of compounds of formula (I), (IA), (IB), (II), (II-1), (II-2), (II-A), (IIA-1), or (IIA-2), R 3 For -H. In the implementation scheme, R 6 For -H. In the implementation scheme, R 3 and R 6 All are -H.

[0261] In embodiments of compounds of formula (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’ Each is -H.

[0262] In embodiments of compounds of formula (I), (II), (IA), (IB), (II-1), (II-2), (II-A), (IIA-1), or (IIA-2), L comprises a cholesterol moiety or at least one fatty acid chain comprising at least 13 carbons. In embodiments, L comprises a cholesterol moiety. In embodiments, L comprises at least one fatty acid chain comprising at least 13 carbons. In embodiments, L comprises at least two fatty acid chains comprising at least 15 carbons. In embodiments, L comprises at least one C 17 Fatty acid chain. In the implementation, L contains at least two C's. 17 Fatty acid chain. In the embodiment, L contains at least one component independently selected from C. 17 Alkyl or C 17 An alkenyl fatty acid chain. In an embodiment, L comprises at least two independently selected from C. 17 Alkyl or C 17 An alkenyl fatty acid chain. In the embodiment, C 17 The fatty acid chain is derived from stearic acid or oleic acid. In the implementation scheme, C 17 The fatty acid chain is derived from stearic acid. In the implementation scheme, C 17 The fatty acid chain is derived from oleic acid fatty acid. In the embodiment, L contains two C atoms derived from stearic acid. 17 Fatty acid chains.

[0263] In embodiments of compounds of formula (I), (IA), (IB), (II), (II-1), (II-2), (II-A), (IIA-1), or (IIA-2), L is selected from fatty acid chain, -alkylene-C(O)-W, -alkylene-OC(O)-W, -alkylene-N-(alkylene-C(O)-NR) 11 -alkylene-NR 11 The group consists of -C(O)-W)2 and -alkylene-N-(alkylene-C(O)-W)2.

[0264] In embodiments of compounds of formula (I), (IA), (IB), (II), (II-1), (II-2), (II-A), (IIA-1), or (IIA-2), L is selected from 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 group consists of -C(O)-W)2 and -CH2CH2-N-(CH2CH2-C(O)-W)2.

[0265] In embodiments of compounds of formula (I), (IA), (IB), (II), (II-1), (II-2), (II-A), (IIA-1), or (IIA-2), L is C 8-30 Fatty acid chain. In the embodiment, L is C 8-30 Alkyl or C 8-30 Alkenyl group. In the embodiment, L is C. 15-20 Alkyl or C 15-20 Alkenyl group.

[0266] In embodiments of compounds of formula (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.

[0267] In embodiments of compounds of formula (I), (IA), (IB), (II), (II-1), (II-2), (II-A), (IIA-1), or (IIA-2), L is -alkylene-C(O)-W. In embodiments, L is -C 1-6 Alkylene-C(O)-W. In the embodiments, L is -CH2-C(O)-W.

[0268] In embodiments of compounds of formula (I), (IA), (IB), (II), (II-1), (II-2), (II-A), (IIA-1), or (IIA-2), L is -alkylene-OC(O)-W. In embodiments, L is -C 1-6 Alkylene-OC(O)-W. In the embodiments, L is -CH2-OC(O)-W.

[0269] In embodiments of compounds of formula (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 -C(O)-W)2. In the implementation scheme, L is -C 2-6 alkylene-N-(-C) 2-6 Alkylene-C(O)-NR 11 -C 2-6 Alkylene-NR 11-C(O)-W)2. In the implementation, L is -CH2-CH2-N-(CH2-CH2-C(O)-NR). 11 -CH2-CH2-NR 11 -C(O)-W)2.

[0270] In embodiments of compounds of formula (I), (IA), (IB), (II), (II-1), (II-2), (II-A), (IIA-1), or (IIA-2), L is -alkylene-N-(alkylene-C(O)-W)2. In embodiments, L is -C 1-6 alkylene-N-(C 1-6 Alkylene-C(O)-W)2. In the embodiment, L is...

[0271] -CH2-CH2-N-(CH2-CH2-C(O)-W)2.

[0272] In embodiments of compounds of formula (I), (IA), (IB), (II), (II-1), (II-2), (II-A), (IIA-1), or (IIA-2), L is C 18 Fatty acid chain. In the embodiment, L is C 18 Alkyl or C 18 Alkenyl group. In the embodiments, L is -CH2(CH2CH2)8-CH3.

[0273] In embodiments of compounds of formula (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.

[0274] In embodiments of compounds of formula (I), (IA), (IB), (II), (II-1), (II-2), (II-A), (IIA-1), or (IIA-2), W is C 8-30 Fatty acid chain. In the implementation scheme, W is C 8-30 Alkyl or C 8-30 Alkenyl group. In the embodiment, W is C. 8-30 Alkyl group. In the embodiment, W is C6. 8-30 Alkenyl group. In the embodiment, W is C. 12-18 Fatty acid chain. In the implementation scheme, W is C 12-18 Alkyl or C 12-18 Alkenyl group. In the embodiment, W is C. 12-18 Alkyl group. In the embodiment, W is C6. 12-18Alkenyl group. In the embodiment, W is C. 18 Fatty acid chain. In the implementation scheme, W is C 17 Fatty acid chain. In the implementation scheme, W is C 17 Alkyl or C 17 Alkenyl group. In the embodiment, W is C. 17 Alkyl group. In the embodiment, W is C6. 17 Alkenyl group. 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.

[0275] In embodiments of compounds of formula (I), (IA), (IB), (II), (II-1), (II-2), (II-A), (IIA-1), or (IIA-2), W is:

[0276] In the implementation plan, R X and R X’ Each is an independent fatty acid chain. In the implementation scheme, R X and R X’ Each is an independent fatty acid chain containing at least 15 carbons. In the implementation scheme, R X and R X’ Each is an independent fatty acid chain containing at least 17 carbons. In some embodiments, R X and R X’ Each independently is -C 8-30 Fatty acid chain. In the implementation scheme, R X and R X’ Each independently is -C 8-30 Alkyl or -C 8-30 Alkenyl. In the embodiment, R X and R X’ All are -C 8-30 Alkyl group. In the implementation scheme, R X and R X’ All are -C 8-30 Alkenyl. In the embodiment, R X and R X’ Each independently is C 12-18 Fatty acid chain. In the implementation scheme, R X and R X’ Each independently is -C 12-18 Alkyl or -C 12-18 Alkenyl. In the embodiment, R X and R X’ -C12-18 Alkyl group. In the implementation scheme, R X and R X’ -C 12-18 Alkenyl. In the embodiment, R X and R X’ Each independently is C 17 Fatty acid chain. In the implementation scheme, R X and R X’ Each independently is C 17 Alkyl or C 17 Alkenyl. In the embodiment, R X and R X’ C 17 Alkyl group. In the implementation scheme, R X and R X’ C 17 Alkenyl. In the embodiment, C 17 The chain is independently derived from stearic acid or oleic acid. In the implementation scheme, C 17 The chain is derived from stearic acid. In the implementation scheme, C... 17 The chain is derived from oleic acid. In the implementation plan, R... X and R X’ All are -(CH2CH2)8-CH3.

[0277] In embodiments of compounds of formula (I), (IA), (IB), (II), (II-1), (II-2), (II-A), (IIA-1), or (IIA-2), W is a sterol.

[0278] In embodiments of compounds of formula (I), (IA), (IB), (II), (II-1), (II-2), (II-A), (IIA-1), or (IIA-2), W is cholesterol:

[0279]

[0280] In embodiments of compounds of formula (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 embodiments, W is phosphatidylcholine (PC). In embodiments, W is phosphatidylglycerol (PG). In embodiments, W is phosphatidylserine (PS). In embodiments, W is phosphatidylethanolamine (PE). In embodiments, W is phosphatidic acid (PA). In embodiments, W is lysophosphatidylcholine.

[0281] In embodiments of compounds of formula (I), (IA), (IB), (II), (II-1), (II-2), (II-A), (IIA-1), or (IIA-2), W is:

[0282]

[0283] in

[0284] Y Q1 Y Q2 and Y Q3 Each is independently an alkylene group. In the implementation scheme, Y Q1 C 2-6 Alkylene, and Y Q2 and Y Q3 Each independently is -C 1-3 Alkylene. In the implementation scheme, R X and R X’ Each is independently a fatty acid chain having at least 15 carbons, or in a specific embodiment, R X and R X’ Each is an independent fatty acid chain having at least 17 carbons. In the implementation scheme, R X and R X’ Each independently is C 8-30 Fatty acid chain. In the implementation scheme, R X and R X’ Each independently is C 8-30 Alkyl or C 8-30 Alkenyl. In the embodiment, R X and R X’ Each independently is C 15-30 Alkyl or C 15-30 Alkenyl. In the embodiment, R X and R X’ Each independently is C 15-20 Alkyl or C 15-20 Alkenyl. In the embodiment, R X and R X’ Each independently is C 17 Alkyl or C 17 Alkenyl. In the embodiment, R X and R X’ All are -(CH2CH2)8-CH3.

[0285] In embodiments of compounds of formula (I), (IA), (IB), (II), (II-1), (II-2), (II-A), (IIA-1), or (IIA-2), W is:

[0286]

[0287] In embodiments of compounds of formula (I), (IA), (IB), (II), (II-1), (II-2), (II-A), (IIA-1), or (IIA-2), W is:

[0288] Or its pharmaceutically acceptable salt;

[0289] Where R X and R X’ Each independently is C 8-30 Fatty acid chain. In the embodiment, the fatty acid is saturated. In the embodiment, R X and R X’ Each independently is C 8-30 Alkyl or C 8-30 Alkenyl. In the embodiment, R X and R X’ Each independently is C 15-20 Alkyl or C 15-20 Alkenyl. In the embodiment, R X and R X’ Each independently is C 17 Alkyl or C 17 Alkenyl. In the embodiment, R X and R X’ All are -(CH2CH2)8-CH3.

[0290] In embodiments of compounds of formula (I), (IA), (IB), (II), (II-1), (II-2), (II-A), (IIA-1), or (IIA-2), R 8 and R 9 Each independently as R X or -C(O)-R X ;

[0291] In embodiments of compounds of formula (I), (IA), (IB), (II), (II-1), (II-2), (II-A), (IIA-1), or (IIA-2), R 6 and R 11 Each is independently -H or alkyl.

[0292] In the embodiments, this disclosure provides compounds selected from the group consisting of:

[0293]

[0294]

[0295] Or its stereoisomers (e.g., its α or β end anomers or their tautomers).

[0296] In the embodiments, this disclosure provides compounds selected from the group consisting of:

[0297]

[0298]

[0299] Or its stereoisomers (e.g., its end-group epiisomers or mixtures of its end-group epiisomers).

[0300] In the embodiments, this disclosure provides compounds of formula (I), (IA), (IB), (II), (II-1), (II-2), (IIA), (IIA-1), (IIA-2) or their stereoisomers.

[0301] In the embodiments, this disclosure provides compounds of formula (I), (IA), (IB), (II), (II-1), (II-2), (IIA), (IIA-1), (IIA-2) or their diastereomers or tautomers.

[0302] In the implementation scheme, one or more compounds selected from Table 1 are provided herein.

[0303] In the implementation scheme, one or more pharmaceutically acceptable salts selected from the compounds in Table 1 are provided herein.

[0304] In the implementation scheme, one or more compounds selected from Table 1, or their stereoisomers or pharmaceutically acceptable salts are provided herein.

[0305] Table 1. Compounds

[0306]

[0307]

[0308]

[0309]

[0310]

[0311] Muraminoyl tripeptide phosphatidylethanolamine; N-(N-acetylmuraminoyl)-L-alanyl-D-α-glutamine acyl-N-[(7R)-4-hydroxy-4-oxo-10-oxo-7-[(1-oxohexadecyl)oxy]-3,5,9-trioxa-4-phosphatidocarbacon-1-yl]-L-alanylamide (MTP-a-DPPE or Mifamurtide): Molecular weight: 1238 Daltons. CLogP = 10.59 (uncharged) and 4.80 (negatively charged). Mifamurtide (CAS No. [83461-56-7]) is classified according to literature procedures (e.g., Brundish, DE; Wade, R. (1985) J Label Compd Radiopharm. 22(1):29-35).

[0312] The molecule was prepared using the method described in doi:10.1002 / jlcr.2580220105. Its lipophilicity is relatively low in physiological environments, with a CLogP of 4.80.

[0313] N-Acetylmurayl-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 No. [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, making it unlikely to adequately ensure its stable incorporation into HDL-derived nanoparticles.

[0314] Romotide (CAS No. [78113-36-7]) Molecular weight: 887 Daltons. CLogP = 3.90 (uncharged) and 0.61 (negatively charged), exhibiting lipophilicity (CLogP 0.61 in physiological environment). This compound has low lipophilicity, and the CLogP of the charged molecule is close to 0.

[0315] Murabutide (CAS No. [74817-61-1]) Molecular weight: 549 Daltons. CLogP = -1.53 ​​(uncharged), exhibiting a negative CLogP value. This molecule is hydrophilic because its CLogP value is below 0.

[0316] In an embodiment, the compounds of this disclosure (e.g., one or more of the formulas (I), (IA), (IB), (II), (II-1), (II-2), (IIA), (IIA-1), (IIA-2), or Table 1) activate protein 2 (NOD2) containing a nucleotide-binding oligomerization domain.

[0317] Terminal diastereomers and open / closed ring structures

[0318] In the embodiments, the molecules of this disclosure have a -OH substituent at the terminal epimer of the hemiacetal carbon of the cell wall acyl glycosyl group, that is, when R2 = H, it should be understood that both terminal epimers α and β are included in the compounds of this disclosure.

[0319] Furthermore, in these cases where R2 = H, it is known in the art that such molecules (in an aqueous environment) actually exist in both closed-ring and open-ring isomers. Again, it should be understood that both open-ring and closed-ring isomers are included in the compounds disclosed herein.

[0320] In the following non-limiting examples, the upper structure shows α and β end-group epimers, and the lower structure shows a typical closed-loop structure (left) and an open-loop structure (right) of end-group epimers.

[0321]

[0322] molecular weight

[0323] The molecular weight of the compounds of the present invention is preferably higher than 500 Daltons, higher than 700 Daltons, higher than 950 Daltons, or higher than 1,200 Daltons.

[0324] The molecular weight of the compounds of the present invention is preferably less than 10,000 Daltons, less than 5,000 Daltons, less than 2,500 Daltons, or less than 1,750 Daltons.

[0325] hydrophobic

[0326] In certain embodiments, the compounds disclosed herein are inherently hydrophobic. Hydrophobicity can be estimated by calculating the CLogP value. This can be done in software programs such as, for example, Perkin Elmer's ChemDraw or ChemDraw Professional (v18). The higher the CLogP value of a compound, the more hydrophobic the compound.

[0327] In the embodiments, the CLogP values ​​of the compounds disclosed herein are 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.

[0328] The CLogP value represents the n-octanol / water partition coefficient (Log Po / w) of a molecule and is a calculated value, the opposite of the LogP value (i.e., the value determined experimentally). Therefore, the CLogP value may deviate from the LogP value. However, importantly, the CLogP value is a good comparison of the lipophilicity of a molecule. The CLogP value can be assessed for molecules in both uncharged and charged states. This is the case for molecules with ionizable groups, such as those with carboxylic acid (-COOH) groups or phosphate (-OP(O)OH-O-) groups. At physiological pH (approximately 7.4), these specific groups deprotonate and become charged. Furthermore, in this case, alkylamine groups become charged at physiological pH through protonation.

[0329] At physiological pH, the CLogP value of the molecules of the present invention is less than 20, less than 15, or less than 10. Furthermore, at physiological pH, the CLogP value of the molecules of the present invention is greater than 3, greater than 4, greater than 5, or greater than 5.5.

[0330] Nanobiocomposition

[0331] This document provides nanobiocompositions comprising nanoparticle carriers and one or more compounds disclosed herein (e.g., compounds of formula (I), (IA), (IB), (II), (II-1), (II-2), (IIA), (IIA-1), or (IIA-2) or the compounds in Table 1 as disclosed herein).

[0332] In embodiments, the compounds of this disclosure can be formulated in nanoparticle carriers, which may include, but are not limited to, polyplexes; colloidal dispersions, such as macromolecular complexes, nanocapsules, microspheres, beads, and lipid-based systems, including oil-in-water emulsions, micelles, mixed micelles, liposomes, lipid complexes, lipid nanoparticles, lipid nanocapsules, lipid-like substances, rapidly eliminated lipid nanoparticles (reLNP), microemulsions, and nanoemulsions; HDL-derived nanoparticles; and polymeric nanoparticles, including poly(lactic-co-glycolic acid) copolymer (PLGA) nanoparticles such as PLGA microspheres, poly(lactide) (PLA) nanoparticles, and poly(ε-caprolactone). (PCL) nanoparticles, poly(butyl cyanoacrylate) (PBCA) nanoparticles, dendritic structures, hyperbranched polyglycerol (HPG) nanoparticles, PEG-polyaspartate micelle nanoparticles; cationic polymers, including, for example, poly(L-lysine), polyethyleneimine (PEI), DEAE-glucan, poly(amino ester) (PBAE), and chitosan; cyclodextrin nanoparticles; metal nanoparticles; surfactant-based emulsions; virus-like particles (e.g., particles mainly composed of viral structural proteins but non-infectious or of 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.

[0333] In embodiments, the compounds of this disclosure can be formulated in nanoparticle carriers, such as those whose contents are incorporated herein by reference in US5,567,434, US5,552,157, US5,565,213, US5,738,868, US5,795,587, US10,485,884, US2018 / 0263907, US2016 / 0317647, US2019 / 0290593, US2020 / 0253884, US2020 / 0376146 and WO2018 / 071549.

[0334] In this embodiment, the nanoparticle carrier is a nanoparticle derived from high-density lipoprotein (HDL). The HDL-derived nanoparticles are envisioned as delivery solvents that can, for example, enhance the therapeutic index of small molecule immunomodulatory compounds and / or impart specific delivery to innate immune cells. By imparting targeting specificity to innate immune cells (e.g., myeloid cells, myeloid progenitor cells, and hematopoietic stem cells in the bone marrow, blood, and / or spleen), therapeutic agents encapsulated or introduced into the HDL-derived nanoparticles can be deposited in a concentrated and localized manner. In this embodiment, the HDL-derived nanoparticles contain apoA-I or a peptide mimic of apoA-I. In this embodiment, the HDL-derived nanoparticles contain apoA-I.

[0335] 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 synthesizing apoA-I by recombinant protein expression, for example, in *E. coli* organisms. When expressed in bacteria, apoA-I may comprise an N-terminal methionine or a formyl-methionine. The presence of the methionine group can be assessed by mass spectrometry (MS) methods known in the art. Also, as is known in the art, the position of the methionine in the protein sequence after digestion of apoA-I can be assessed using subsequent analysis of the peptide mixture using MS.

[0336] In the implementation scheme, purification of apoA-I (including any of its variants) may include any method known in the art (e.g., using hydrophobic interaction chromatography, ion exchange columns, precipitation, etc.). The production method may or may not include the use of affinity tags capable of purifying the protein; such tags need to be removed after purification to restore human apoA-I identity.

[0337] In the implementation scheme, the nanoparticles derived from high-density lipoprotein (HDL) contain ApoA-1Milano.

[0338] Suitable apoA-I mimic peptides may have the sequences shown in Table 2 (SEQ ID NOS: 256 to 263 and 342 to 346) or the sequences shown in SEQ ID NOS: 1 to 341.

[0339] Table 2. ApoA-I Simulation

[0340] SEQ ID NO amino acid sequence 256 DWLKAFYDKVAEKLKEAF(18A) 257 <![CDATA[Ac-DWLKAFYDKVAEKLKEAF-NH 2( 2F)]]> 260 <![CDATA[Ac-DWFKAFYDKVAEKFKEAF-NH 2( 4F)]]> 258 <![CDATA[Ac-DWFKAFYDKVAEKLKEAF-NH2(3 F3 )]]> 259 <![CDATA[Ac-DWLKAFYDKVAEKFKEAF-NH2(3F 14 )]]> 261 <![CDATA[Ac-DWLKAFYDKVFEKFKEFF-NH2(5F)]]> 262 <![CDATA[Ac-DWLKAFYDKFFEKFKEFF-NH2(6F)]]> 263 <![CDATA[Ac-DWFKAFYDKFFEKFKEFF-NH2(7F)]]> 342 <![CDATA[Ac-FWLKAFYDKVAEKLKEAF-NH2(3F-1)]]> 343 <![CDATA[Ac-DFLKAFYDKVAEKLKEAF-NH2(3F-2)]]> 344 <![CDATA[Ac-DWFRAFYDKVAEKFREAF-NH 2( 4F-R) q ]]> 345 <![CDATA[Ac-DWFKAFYDRVAERFKEAF-NH2(4F-R′) q ]]> 346 <![CDATA[Ac-DWLXAFYDXVAEXLXEAF-NH2(2F′)]]>

[0341] In one implementation, the apoA-I simulant is DWLKAFYDKVAEKLKEAF (SEQ ID NO. 256). In another implementation, the apoA-I simulant is Ac-DWLKAFYDKVAEKLKEAF-NH2 (SEQ ID NO. 257). In yet another implementation, the apoA-I simulant is Ac-DWFKAFYDKVAEKFKEAF-NH2 (SEQ ID NO. 260).

[0342] 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 this disclosure comprise one or more phospholipids. All phospholipids with chain lengths ranging 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 hemolysin are contemplated for use in the nanoparticles described herein. Furthermore, other synthetic variants and variants having other phospholipid head groups are also contemplated. In embodiments, the HDL-derived nanoparticles comprise phospholipids. In embodiments, the HDL-derived nanoparticles comprise phospholipids and hemolysin.

[0343] Non-limiting examples of phospholipids that can be used in this composition include phosphatidylcholine (PC), phosphatidylglycerol (PG), phosphatidylserine (PS), and phosphatidylethanolamine (PE). In embodiments, phosphatidic acid / ester (PA) may be used.

[0344] In the implementation scheme, the phospholipid or lysolipin is one or more of the following: DDPC

[0345] CAS-3436-44-0 1,2-Didecanoyl-sn-glycerol-3-phosphocholine, DEPA-NA

[0346] CAS-80724-31-8 1,2-Dishuranoyl-sn-glycerol-3-phosphate (sodium salt), DEPC

[0347] CAS-56649-39-9 1,2-Disorhodo-sn-glycerol-3-phosphate choline, DEPE CAS-988-07-2 1,2-Disorhodo-sn-glycerol-3-phosphate ethanolamine, DEPG-NA 1,2-Disorhodo-sn-glycerol-3-phosphate-rac-(l-glycerol) (sodium salt), DLOPC CAS-998-06-11,2-Dilinoleoyl-sn-glycerol-3-phosphate choline, DLPA-NA 1,2-Dilauroyl-sn-glycerol-3-phosphate (sodium salt), DLPC CAS-18194-25-7 1,2-Dilauroyl-sn-glycerol-3-phosphate choline, DLPE 1,2-Dilauroyl-sn-glycerol-3-phosphate ethanolamine, DLPG-NA 1,2-Dilauroyl-sn-glycerol-3-phosphate-rac-(l-glycerol) (sodium salt), DLPG-NH4 1,2-Dilauroyl-sn-glycerol-3-phosphate-rac-(l-glycerol) (ammonium salt), DLPS-NA 1,2-Dilauroyl-sn-glycerol-3-phosphoserine (sodium salt), DMPA-NA

[0348] CAS-80724-3 1,2-Dimyristoyl-sn-glycerol-3-phosphate (sodium salt), DMPC

[0349] CAS-18194-24-6 1,2-Dimyristicoyl-sn-glycerol-3-phosphocholine, DMPE

[0350] CAS-988-07-2 1,2-Dimyristoyl-sn-glycerol-3-phosphate ethanolamine, DMPG-NA

[0351] CAS-67232-80-8 1,2-Dimyristicoyl-sn-glycerol-3-phosphate-rac-(l-glycerol) (sodium salt), DMPG-NH4 1,2-Dimyristicoyl-sn-glycerol-3-phosphate-rac-(l-glycerol) (ammonium salt), DMPG-NH4 / NA 1,2-Dimyristicoyl-sn-glycerol-3-phosphate-rac-(l-glycerol) (sodium / ammonium salt), DMPS-NA 1,2-Dimyristicoyl-sn-glycerol-3-phosphoserine (sodium salt), DOPA-NA 1,2-dioleoyl-sn-glycerol-3-phosphate (sodium salt), DOPCCAS-4235-95-4 1,2-dioleoyl-sn-glycerol-3-phosphocholine, DOPE CAS-4004-5-1 1,2-Dioleoyl-sn-glycerol-3-phosphate ethanolamine, DOPG-NA CAS-62700-69-0 1,2-Dioleoyl-sn-glycerol-3-phosphate-rac-(l-glycerol) (sodium salt), DOPS-NA CAS-70614-14-1 1,2-Dioleoyl-sn-glycerol-3-phosphoserine (sodium salt), DPPA-NA CAS-71065-87-7 1,2-Dipalmitoyl-sn-glycerol-3-phosphate (sodium salt), DPPC CAS-63-89-8 1,2-Dipalmitoyl-sn-glycerol-3-phosphate choline, DPPE CAS-923-61-5 1,2-Dipalmitoyl-sn-glycerol-3-phosphate ethanolamine, DPPG-NA CAS-67232-81-9 1,2-Dipalmitoyl-sn-glycerol-3-phosphate-rac-(l-glycerol) (sodium salt), DPPG-NH4 CAS-73548-70-6; 1,2-Dipalmitoyl-sn-glycerol-3-phosphate-rac-(l-glycerol) (ammonium salt), DPPS-NA; 1,2-Dipalmitoyl-sn-glycerol-3-phosphoserine (sodium salt), DSPA-NA CAS-108321-18-2; 1,2-Distearayoyl-sn-glycerol-3-phosphate (sodium salt), DSPC CAS-816-94-4; 1,2-Distearayoyl-sn-glycerol-3-phosphocholine, DSPE CAS-1069-79-0; 1,2-Distearayoyl-sn-glycerol-3-phosphoethanolamine, DSPG-NA CAS-67232-82-0 1,2-Distearyl-sn-glycerol-3-phosphate-rac-(l-glycerol) (sodium salt), DSPG-NH4 CAS-108347-80-4 1,2-Distearyl-sn-glycerol-3-phosphate-rac-(l-glycerol) (ammonium salt), DSPS-NA1,2-Disteasay-sn-glycerol-3-phosphoserine (sodium salt), EPC egg yolk PC, HEPC hydrogenated egg yolk PC, HSPC hydrogenated soybean PC, LYSOPC MYRISTIC CAS-18194-24-6L-myristoyl-sn-glycerol-3-phosphocholine, LYSOPC ALMITIC CAS-17364-16-8L-palmitoyl-sn-glycerol-3-phosphocholine, LYSOPC STEARIC CAS-19420-57-6L-stearoyl-sn-glycerol-3-phosphocholine, sphingomyelin, MPPC l-myristoyl-2-palmitoyl-sn-glycerol-3-phosphocholine, MSPC l-myristoyl-2-stearoyl-sn-glycerol-3-phosphocholine, PMPC l-palmitoyl-2-myristoyl-sn-glycerol-3-phosphocholine, POPC CAS-26853-31-6l-palmitoyl-2-oleoyl-sn-glycerol-3-phosphocholine, POPE l-palmitoyl-2-oleoyl-sn-glycerol-3-phosphoethanolamine, POPG-NACAS-81490-05-3l-palmitoyl-2-oleoyl-sn-glycerol-3[phosphate-rac-(l-glycerol)] (sodium salt), PSPC 1-palmitoyl-2-stearoyl-sn-glycerol-3-phosphocholine, SMPC l-stearoyl-2-myristoyl-sn-glycerol-3-phosphocholine, SOPC l-stearoyl-2-oleoyl-sn-glycerol-3-phosphocholine, SPPC 1-stearoyl-2-palmitoyl-sn-glycerol-3-phosphocholine. In some preferred embodiments, specific non-limiting examples of phospholipids include: dimyristoyl phosphatidylcholine (DMPC), soybean lecithin, dipalmitoyl phosphatidylcholine (DPPC), distearyl phosphatidylcholine (DSPC), dilauryl phosphatidylcholine (DLPC), dioleoyl phosphatidylcholine (DOPC), dilauryl phosphatidylglycerol (DLPG), dimyristoyl phosphatidylglycerol (DMPG), dipalmitoyl phosphatidylglycerol (DPPG), and distearyl phosphatidylglycerol (DSPG). Dioleoylphosphatidylglycerol (DOPG), dimyristoylphosphatidylglycerol (DMPA), dimyristoylphosphatidylglycerol (DMPA), dipalmitoylphosphatidylglycerol (DPPA), dimyristoylphosphatidylethanolamine (DMPE), dipalmitoylphosphatidylethanolamine (DPPE), dimyristoylphosphatidylserine (DMPS), dipalmitoylphosphatidylserine (DPPS), dipalmitoylsphingomyelin (DPSP), distearateylsphingomyelin (DSSP), and mixtures thereof.

[0352] In the implementation scheme, the phospholipid is 1,2-dimyristoyl-sn-glycerol-3-phosphatidylcholine (DMPC), and the lysosomal lipid is 1-myristoyl-2-hydroxy-sn-glycerol-phosphocholine (MHPC).

[0353] In the implementation scheme, the phospholipid is 1-palmitoyl-2-oleoyl-sn-glycerol-3-phosphocholine (POPC), and the hemolysin is 1-palmitoyl-2-hydroxy-sn-glycerol-3-phosphocholine (PHPC).

[0354] In embodiments, when the composition comprises two or more types of lipids (e.g., phospholipids or lysolipin) (or is substantially composed of or consists of), the weight ratio of the two types of phospholipids ranges from about 1:10 to about 10:1, including 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 therein.

[0355] In the implementation scheme, the HDL-derived nanoparticles comprise DMPC and MHPC, and the weight ratio of DMPC to MHPC can 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 can 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 therein.

[0356] In the implementation scheme, the HDL-derived nanoparticles comprise POPC and PHPC, and the weight ratio of POPC to PHPC can 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 can 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.

[0357] In the embodiments, the phospholipids in the nanoparticles of this disclosure comprise (or consist substantially of) a mixture of diacyl-phospholipids and monoacyl-phospholipids / lysolipin.

[0358] In one embodiment, the high-density lipoprotein (HDL) derived nanoparticles comprise apoA-I or a peptide mimic of apoA-I and phospholipids. In another embodiment, the high-density lipoprotein (HDL) derived nanoparticles comprise apoA-I or a peptide mimic of apoA-I, phospholipids, and compounds of formulas (I), (II), (II-1), (II-2), (II-A), (IIA-1), or (IIA-2).

[0359] In one embodiment, the high-density lipoprotein (HDL) derived nanoparticles comprise i) apoA-I or a peptide mimic of apoA-I; ii) phospholipids; iii) lysed lipids; and iv) cholesterol. In another embodiment, the high-density lipoprotein (HDL) derived nanoparticles comprise i) apoA-I or a peptide mimic of apoA-I; ii) phospholipids; iii) lysed lipids; iv) cholesterol and a compound of formula (I), (II), (II-1), (II-2), (II-A), (IIA-1), or (IIA-2).

[0360] In one embodiment, the nanoparticles derived from high-density lipoprotein (HDL) comprise i) apoA-I or a peptide mimic of apoA-I; ii) phospholipids; and iii) cholesterol. In another embodiment, the nanoparticles derived from high-density lipoprotein (HDL) comprise i) apoA-I or a peptide mimic 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).

[0361] In the embodiments, the nanoparticles derived from high-density lipoprotein (HDL) comprise i) apoA-I or a peptide mimic of apoA-I; ii) phospholipids; iii) lysed lipids; iv) a hydrophobic matrix core; and v) cholesterol. In the embodiments, the nanoparticles derived from high-density lipoprotein (HDL) comprise i) apoA-I or a peptide mimic of apoA-I; ii) phospholipids; iii) lysed lipids; 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).

[0362] In the embodiments, the nanoparticles derived from high-density lipoprotein (HDL) comprise i) apoA-I or a peptide mimic of apoA-I; ii) phospholipids; iii) lysed lipids; iv) triglycerides; v) cholesterol and compounds of formula (I), (II), (II-1), (II-2), (II-A), (IIA-1), or (IIA-2).

[0363] In implementation methods, the structure and properties (e.g., particle size, rigidity, viscosity, loading, etc.) of HDL-derived nanoparticles can be altered by introducing a hydrophobic matrix. As used herein, a hydrophobic matrix refers to the core or filler or structural modifier of a nanobiologic. Non-limiting examples of suitable hydrophobic matrix molecules include triglycerides, fatty acid esters, hydrophobic polymers, sterol esters, or combinations thereof.

[0364] For example, the inclusion of one or more triglycerides and / or one or more polymers in the nanoparticles disclosed herein facilitates the tuning of nanoparticle size (e.g., from about 10 nm to over 100 nm) and shape (from disc-shaped to spherical). Conversely, the size, rigidity, and viscosity of HDL-derived nanoparticles may also affect loading and biodistribution. In non-limiting examples, HDL-derived nanoparticles containing phospholipids and apoA-I can have diameters from about 10 nm to about 50 nm, and the addition of hydrophobic matrix molecules (e.g., triglycerides) causes HDL-derived nanoparticles to swell from a minimum of about 10 nm to at least about 30 nm. Adding more triglycerides can further increase the diameter of 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 therein.

[0365] It is anticipated that any suitable synthetic or natural fatty acid or fatty acid ester known in the art may be used in the HDL-derived nanoparticles of this disclosure. Non-limiting examples of fatty acids used include: arachidonic acid, oleic acid, arachidic acid, lauric acid, sad, decanoic acid, myristic acid, palmitic acid, stearic acid, linoleic acid, linolenic acid, didecanoate, tridecanoic acid esters, glyceryl monofatty acid esters, glyceryl dilaurate, 1-Sunsoft 767, laurocapramone (1-dodecyl-aza-cycloheptane-2-one), acylcarnitine, acylcholine, or C1-C 10 arrcostab (e.g., isopropyl myristate IPM), monoglycerides, diglycerides, or pharmaceutically acceptable salts thereof.

[0366] It is anticipated that any suitable synthetic or natural triglycerides known in the art may be used in the HDL-derived nanoparticles of this disclosure. Non-limiting examples of triglycerides used include: tricaprylic acid glyceride, tristearate glyceride, trioleic acid glyceride, tripalmitic acid glyceride, 1,2-dipalmitoyl glyceride, 1,3-dipalmitoyl glyceride, 1-palmitoyl-3-stearic acid-2-oleic acid glyceride, 1-palmitoyl-2-stearic acid-3-oleic acid glyceride, 2-palmitoyl-1-stearic acid-3-oleic acid glyceride, trilinoleic acid glyceride, 1,2-dipalmitoyllinoleic acid glyceride, 1-palmitoyl-dilinoleic acid glyceride, 1,2-diacetylpalmitoyl glyceride, 1,2-distearate-oleic acid glyceride, 1,3-distearate-oleic acid glyceride, trimyristate glyceride, trilaurate glyceride, and combinations thereof. Suitable triglycerides may be added to the composition in pure form. Alternatively, oils and / or processed oils containing suitable triglycerides 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 seed oil, whale oil, soybean oil, peanut oil, flaxseed oil, rosin oil, and combinations thereof.

[0367] One or more hydrophobic polymers may be selected from the group consisting of polymers approved for human use (i.e., biocompatible and FDA approved). Such polymers include, for example, but not limited to, the following polymers, derivatives of such polymers, copolymers, block copolymers, branched polymers, and polymer blends: polyolefin dicarboxylate, polyanhydride, poly(aspartic acid), polyamide, polybutylene succinate (PBS), polybutylene succinate-adipate copolymer (PBSA), poly(ε-caprolactone) (PCL), polycarbonates including polyalkylene carbonate (PC), polyesters including aliphatic polyesters and polyester-amides, and polyethylene succinate. (PES), polyglycolic acid (PGA), polyimide and polyalkylimide (Pl, PAI), polylactide (PLA (polylactic acid), PLLA, PDLLA), polylactic acid-glycolic acid copolymer (PLGA), poly(l-lysine), polymethacrylate, polypeptides, polyorthoesters, poly-p-dioxanone (PPDO), (hydrophobic) modified polysaccharides, polysiloxanes and poly-alkyl-siloxanes, polyurea, polyurethane, and polyvinyl alcohol, and biodegradable polyalkyl-cyanoacrylates.

[0368] In embodiments of the HDL-derived nanoparticles disclosed herein, the addition of cholesterol to the nanoparticle carrier stabilizes the composition and improves capture efficiency. Typically, the HDL-derived nanoparticles contain about 1 mol% to about 100 mol% cholesterol relative to phospholipids (e.g., relative to DMPC), including 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%, and about 22 mol%. The range and values ​​are approximately 23 mol%, approximately 24 mol%, approximately 25 mol%, approximately 26 mol%, approximately 27 mol%, approximately 28 mol%, approximately 29 mol%, approximately 30 mol%, approximately 35 mol%, approximately 40 mol%, approximately 45 mol%, approximately 50 mol%, approximately 55 mol%, approximately 60 mol%, approximately 65 mol%, approximately 70 mol%, approximately 75 mol%, approximately 80 mol%, approximately 85 mol%, approximately 90 mol%, approximately 95 mol%, to approximately 100 mol% (i.e., a 1:1 mol / mol mixture of cholesterol and phospholipids (e.g., DMPC), including all ranges and values ​​therein. In one embodiment, the HDL-derived nanoparticles contain approximately 1 mol% to approximately 30 mol% cholesterol. In another embodiment, the HDL-derived nanoparticles contain approximately 15 mol% to approximately 25 mol% cholesterol relative to phospholipids. In yet another embodiment, the HDL-derived nanoparticles contain approximately 20 mol% cholesterol relative to phospholipids. In one embodiment, the HDL-derived nanoparticles contain about 10 mol% to about 35 mol% cholesterol relative to phospholipids. In another embodiment, the HDL-derived nanoparticles contain about 15 mol% to about 30 mol% cholesterol relative to phospholipids. In yet another embodiment, the HDL-derived nanoparticles contain about 15 mol% to about 25 mol% cholesterol relative to phospholipids. In yet another embodiment, the HDL-derived nanoparticles contain about 28 mol% to about 23 mol% cholesterol relative to phospholipids. In yet another embodiment, the HDL-derived nanoparticles contain about 20 mol% to about 27 mol% cholesterol relative to phospholipids.

[0369] In the implementation scheme, the HDL-derived nanoparticles are cholesterol-free. In the implementation scheme, the cholesterol:phospholipid molar ratio in the HDL-derived nanoparticles is approximately 0:1, approximately 0.025:1, approximately 0.05:1, approximately 0.075:1, approximately 0.1:1, approximately 0.125:1, approximately 0.15:1, approximately 0.175:1, approximately 0.2:1, approximately 0.225:1, approximately 0.25:1, approximately 0.275:1, approximately 0.3:1, approximately 0.325:1, approximately 0.35:1, approximately 0.375:1, approximately 0.4:1, approximately 0.425:1, approximately 0.45:1, approximately 0.475:1, or approximately 0.5:1, including all values ​​in between. In the embodiments, the molar ratio of cholesterol to phospholipids ranges from about 0:1 to about 0.5:1, including 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, and all ranges therein. In the embodiments, the molar ratio of cholesterol to phospholipids ranges from about 0.05:1 to about 0.25:1. In the embodiments, the molar ratio of cholesterol is about 0.2:1.

[0370] In one embodiment, the HDL-derived nanoparticles contain one or more phospholipids and cholesterol in a molar ratio ranging from about 1:0.05 to about 1:0.25. In another embodiment, the HDL-derived nanoparticles contain one or more phospholipids and cholesterol in a molar ratio of about 1:0.2.

[0371] In the embodiments, the weight percentage of cholesterol ranges from about 0% (w / w) to about 15% (w / w) of the nanoparticles, lipids, or composition, including 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), and so on. (w / w), approximately 7.5% (w / w), approximately 8% (w / w), approximately 8.5% (w / w), approximately 9% (w / w), approximately 9.5% (w / w), approximately 10% (w / w), approximately 10.5% (w / w), approximately 11% (w / w), approximately 11.5% (w / w), approximately 12% (w / w), approximately 12.5% ​​(w / w), approximately 13% (w / w), approximately 13.5% (w / w), approximately 14% (w / w), approximately 14.5% (w / w) and approximately 15% (w / w). In the embodiments, the weight percentage of cholesterol ranges from about 0% (w / w) to about 15% (w / w) of the nanoparticles, lipids, or composition, including 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), and so on. The percentages are approximately 7.5% (w / w), approximately 8% (w / w), approximately 8.5% (w / w), approximately 9% (w / w), approximately 9.5% (w / w), approximately 10% (w / w), approximately 10.5% (w / w), approximately 11% (w / w), approximately 11.5% (w / w), approximately 12% (w / w), approximately 12.5% ​​(w / w), approximately 13% (w / w), approximately 13.5% (w / w), approximately 14% (w / w), approximately 14.5% (w / w), and approximately 15% (w / w). In embodiments, the weight percentage is the weight percentage of cholesterol relative to phospholipids. In embodiments, the weight percentage of cholesterol ranges from approximately 1% to 10% (w / w%) of cholesterol in the composition. The weight percentage of cholesterol ranges from approximately 2% to 8% (w / w%) of cholesterol in the composition. In embodiments, the weight percentage of cholesterol ranges from approximately 3.5% to 7.5% (w / w%) of cholesterol in the composition. In one embodiment, the weight percentage of cholesterol ranges from about 5 to 10% (w / w%) of cholesterol in the composition. In another embodiment, the weight percentage of cholesterol is about 3.6% (w / w%) of cholesterol in the composition. In yet another embodiment, the weight percentage of cholesterol is about 7.2% (w / w%) of cholesterol in the composition. In yet another embodiment, the weight percentage of cholesterol is about 5.9% (w / w%) of cholesterol in the composition.

[0372] In the implementation scheme, the size and cycle time of the nanoparticles can be adjusted, for example, by controlling the ratio of lipids to APOAl and the ratio of lipids to polymers or lipids to triglycerides.

[0373] In the implementation scheme, the HDL-derived nanoparticles comprise a phospholipid:apoA-I or an apoA-I analog in a ratio of approximately 5:1 to 1000:1 (e.g., on a molar basis), including approximately 5:1, approximately 10:1, approximately 20:1, approximately 30:1, approximately 40:1, approximately 50:1, approximately 60:1, approximately 70:1, approximately 80:1, approximately 90:1, approximately 100:1, approximately 110:1, approximately 120:1, approximately 130:1, approximately 140:1, approximately 150:1, approximately 160:1, approximately 170:1, approximately 180:1, and approximately 190:1. Approximately 200:1, Approximately 210:1, Approximately 220:1, Approximately 230:1, Approximately 240:1, Approximately 250:1, Approximately 260:1, Approximately 270:1, Approximately 280:1, Approximately 290:1, Approximately 300:1, Approximately 310:1, Approximately 320:1, Approximately 330:1, Approximately 340:1, Approximately 350:1, Approximately 360:1, Approximately 370:1, Approximately 380:1, Approximately 390:1, Approximately 400:1, Approximately 410:1, Approximately 420:1, Approximately 430:1, Approximately 440:1, Approximately 450:1, Approximately 460:1, Approximately 470 :1, Approximately 480:1, Approximately 490:1, Approximately 500:1, Approximately 510:1, Approximately 520:1, Approximately 530:1, Approximately 540:1, Approximately 550:1, Approximately 560:1, Approximately 570:1, Approximately 580:1, Approximately 590:1, Approximately 600:1, Approximately 610:1, Approximately 620:1, Approximately 630:1, Approximately 640:1, Approximately 650:1, Approximately 660:1, Approximately 670:1, Approximately 680:1, Approximately 690:1, Approximately 700:1, Approximately 710:1, Approximately 720:1, Approximately 730:1, Approximately 740:1, Approximately 7 The ratios are approximately 50:1, 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 ​​therein. In embodiments, the HDL-derived nanoparticles comprise a phospholipid:apoA-I or an apoA-I analogue in a ratio of approximately 10:1 to 1000:1 (e.g., molar). In one embodiment, the HDL-derived nanoparticles comprise a phospholipid:apoA-I ratio of approximately 70:1 to 125:1 (e.g., molar). In another embodiment, the HDL-derived nanoparticles comprise an apoA-I mimicry in a ratio of approximately 5:1 to 10:1 (e.g., molar).

[0374] In the implementation scheme, the HDL-derived nanoparticles contain approximately 2:1 to 3:1 (by weight) of phospholipid:apoA-I or an analogue of apoA-I.

[0375] In the embodiments, the HDL-derived nanoparticles contain 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), including 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 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 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%. 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% (a 1:1 mol / mol mixture of the compound and phospholipid (e.g., DMPC), including all ranges and values ​​therein. In embodiments, the HDL-derived nanoparticles contain about 10 mol% to about 30 mol% of the compound of Formula I relative to the phospholipid. In embodiments, the HDL-derived nanoparticles contain about 12 mol% to about 25 mol% of the compound relative to the phospholipid.

[0376] In the implementation scheme, the size of the nanoparticles ranges from about 5 nm to about 500 nm in diameter, including 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, about 160 nm, about 170 nm, about 180 nm, about 190 nm, about 200 nm, and about 200 nm. The nanoparticle size ranges from 10 nm, approximately 220 nm, approximately 230 nm, approximately 240 nm, approximately 250 nm, approximately 260 nm, approximately 270 nm, approximately 280 nm, approximately 290 nm, approximately 300 nm, approximately 310 nm, approximately 320 nm, approximately 330 nm, approximately 340 nm, approximately 350 nm, approximately 360 nm, approximately 370 nm, approximately 380 nm, approximately 390 nm, approximately 400 nm, approximately 410 nm, approximately 420 nm, approximately 430 nm, approximately 440 nm, approximately 450 nm, approximately 460 nm, approximately 470 nm, approximately 480 nm, approximately 490 nm to approximately 500 nm, including all ranges and values ​​therebetween. In one embodiment, the nanoparticle size is less than approximately 50 nm. In another embodiment, the nanoparticle size is approximately 50 nm to approximately 100 nm, or approximately 5 nm to approximately 30 nm. In yet another embodiment, the nanoparticle size is measured by dynamic light scattering (DLS). In one embodiment, nanoparticles with a long blood half-life and a small size (<50 nm) can be used to target immune cells entering tissues with restricted circulation. In another embodiment, nanoparticles with a short blood half-life and a large size (approximately 100 nm) can be used to target immune cells in well-perfused tissues. These tissues include the spleen, liver, kidneys, lungs, and bone marrow.

[0377] In one embodiment, the HDL-derived nanoparticles are disk-shaped. In another embodiment, the HDL-derived nanoparticles are spherical. In yet another embodiment, the morphology of the HDL-derived nanoparticles is visualized using transmission electron microscopy (TEM).

[0378] In one embodiment, the length of the HDL-derived nanoparticles is from about 5 to about 100 nm, including lengths of 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, including all ranges and values ​​therein. In another embodiment, the length of the HDL-derived nanoparticles is from about 10 nm to 80 nm. In yet another embodiment, the length of the HDL-derived nanoparticles is from about 15 nm to 50 nm. In yet another embodiment, the HDL-derived nanoparticles are longer than about 10 nm or longer than about 15 nm. In the embodiments, the thickness of the HDL-derived nanoparticles is from about 1 nm to 10 nm, including 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 ​​therein. In the embodiments, the thickness of the HDL-particles is about 1 to 10 nm, or 2 to 7 nm, or 3 to 6 nm. In the embodiments, the dimensions (e.g., length and thickness) are recorded by cryo-TEM. In the embodiments, the HDL-particles have a worm-like morphology using cryo-TEM.

[0379] In one embodiment, the HDL-derived nanoparticles are disk-shaped with a diameter between about 5 nm and about 50 nm (e.g., as measured by dynamic light scattering (DLS), including about 5 nm, about 10 nm, about 15 nm, about 20 nm, about 25 nm, about 30 nm, about 40 nm and about 50 nm, including all subranges and values ​​therein. In another embodiment, the diameter of the nanodisc is about 5 nm to about 30 nm.

[0380] In the implementation scheme, the HDL-derived nanoparticles are spherical in shape with a diameter between about 10 nm and about 400 nm (e.g., as measured by dynamic light scattering (DLS)), including 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, and about 160 nm. The nanospheres have diameters ranging from approximately 170 nm, 180 nm, 190 nm, 200 nm, 210 nm, 220 nm, 230 nm, 240 nm, 250 nm, 260 nm, 270 nm, 280 nm, 290 nm, 300 nm, 310 nm, 330 nm, 340 nm, 350 nm, 360 nm, 370 nm, 380 nm, and 390 nm to approximately 400 nm, including all values ​​and ranges therebetween. In one embodiment, the diameter of the nanospheres is between approximately 15 nm and approximately 250 nm. In another embodiment, the diameter of the nanospheres is between approximately 30 nm and approximately 100 nm.

[0381] The stability of HDL-derived nanoparticles can be assessed by performing DLS measurements. In this embodiment, using DLS, HDL-derived nanoparticles are stabilized for, for example, at least about 1 week, or at least about 2 weeks, or at least about 5 weeks.

[0382] In one embodiment, the nanobiocomposition promotes a hyperresponsive innate immune response in patients in need. In another embodiment, the hyperresponsive innate immune response is promoted for at least about 7 to about 30 days. In yet another embodiment, the hyperresponsive innate immune response is promoted for at least 30 to 100 days. In yet another embodiment, the hyperresponsive innate immune response is promoted for more than 100 days and up to 3 years. In yet another embodiment, the nanobiocomposition is administered once, and the hyperresponsive innate immune response is promoted for at least 30 days. In yet another embodiment, in a multiple-dose regimen, the nanobiocomposition is administered at least once daily, and the hyperresponsive innate immune response is promoted for at least 30 days.

[0383] Manufacturing methods can produce uniformly sized HDL-derived nanoparticles, or a mixture of non-uniformly sized HDL-derived nanoparticles by unfiltering or by preparing a series of HDL-derived nanoparticles of different sizes and recombining them in subsequent manufacturing steps. Larger HDL-derived nanoparticles can accommodate more drug. However, larger sizes, such as >120 nm, can limit, prevent, or slow the diffusion of HDL-derived nanoparticles into the tissues of the patient being treated. Smaller HDL-derived nanoparticles cannot each contain the same amount of drug, but can reach the bone marrow, blood, or spleen, or other local tissues affected by trained immunity, such as myeloid cells, myeloid progenitor cells, and hematopoietic stem cells in the bone marrow, blood, and / or spleen (biodistribution).

[0384] The use of mixtures of non-uniformly sized nanoparticles in a single dose or regimen can result in an immediate reduction of innate immune hyperresponsiveness, and simultaneously produce a persistent, long-term reduction of innate immune hyperresponsiveness that can last for days, weeks, months, and years. Nanobiopharmaceuticals reverse, alter, or remodulate the metabolic, epigenetic, and inflammasome pathways of hematopoietic stem cells (HSCs), common myeloid progenitor cells (CMPs), and myeloid cells such as monocytes, macrophages, and other short-lived circulating cells.

[0385] In the implementation scheme, the maximum loading capacity of the HDL-derived nanoparticles can be determined by dividing the internal volume of the HDL-derived nanoparticles by the volume of the drug-loaded spheres.

[0386] Particles: Assume 100nm spherical particles with 2.2nm-3.0nm phospholipid walls produce an interior with a diameter of 94nm and a volume (L)@4 / 3n(r)3.

[0387] Drug: Assume that the STIMULATOR is a cylinder of 12×12×35 angstroms or 1.2×1.2×3.5 nm. Multiple drug molecules cylinders, such as 7 or 9, can be assumed to be spheres with a diameter of 3.5 nm and a radius of 1.75 nm. Volume (small)@4 / 3n(r)3.

[0388] Maximum load capacity (calculated): 100nm particle - 487k 3.5nm sphere.

[0389] preparation

[0390] When used as pharmaceutical agents, the compounds of this disclosure and the HDL-derived nanoparticles are typically administered in the form of pharmaceutical compositions. Such compositions can be prepared in a manner known in the pharmaceutical industry and comprise at least one active compound. In embodiments, the pharmaceutical composition comprises the nanobiocomposite of this disclosure and a pharmaceutically acceptable carrier.

[0391] Typically, the compounds of this invention are administered in pharmaceutically effective amounts. The actual amount of compound administered will usually be determined by a physician based on relevant circumstances, including the condition to be treated, the route of administration chosen, the actual compound administered, the individual patient's age, weight and response, and the severity of the patient's symptoms.

[0392] The pharmaceutical compositions of the present invention can be administered via a variety of routes, including oral, rectal, intraocular, percutaneous, subcutaneous, intravenous, intramuscular, intraperitoneal, intradermal, direct delivery into cerebrospinal fluid, intratracheal, and intranasal administration. Depending on the intended delivery route, the compounds of the present invention are preferably formulated as injectable or oral compositions, or as ointments, lotions, or patches for transdermal administration. In embodiments, the compositions are administered intravenously or intra-arterially.

[0393] Compositions intended for oral administration may be in the form of bulk liquid solutions, suspensions, or bulk powders. However, more commonly, compositions are presented in unit dosage forms to facilitate accurate dosing. The term "unit dosage form" refers to a physically separated unit suitable for use as a unit dose for human subjects and other mammals, each unit containing a predetermined amount of active substance calculated to produce the desired therapeutic effect, along with suitable pharmaceutical excipients. Typical unit dosage forms include pre-filled, pre-measured ampoules or syringes of liquid compositions, or, in the case of solid compositions, pills, tablets, or capsules. In such compositions, the compound as described herein is typically a minor component (about 0.1 to about 50% by weight, or preferably about 1 to about 40% by weight), with the remainder being various solvents or carriers and processing aids that contribute to the formation of the desired dosage form.

[0394] Liquid forms suitable for oral administration may include suitable aqueous or non-aqueous solvents having buffers, suspending and dispersing agents, colorants, and flavoring agents. Solid forms may include compounds with properties similar to any of the following: binders, such as microcrystalline cellulose, tragacanth gum, or gelatin; excipients, such as starch or lactose; disintegrants, such as alginate, Primogel, or corn starch; lubricants, such as magnesium stearate; gliding agents, such as colloidal silica; sweeteners, such as sucrose or saccharin; or flavoring agents, such as peppermint, methyl salicylate, or orange flavoring.

[0395] Injectable compositions are typically based on injectable sterile saline or phosphate-buffered saline or other injectable carriers known in the art. As previously mentioned, the active compound in such compositions is usually a minor component, often about 0.05 to 10% by weight, with the remainder being the injectable carrier, etc.

[0396] Transdermal compositions are typically formulated as topical ointments or creams containing an active ingredient in an amount ranging from about 0.01 to about 20% by weight, preferably from about 0.1 to about 20% by weight, more preferably from about 0.1 to about 10% by weight, and more preferably from about 0.5 to about 15% by weight. When formulated as an ointment, the active ingredient is typically combined with a paraffin base or a water-miscible ointment base. Alternatively, the active ingredient may be formulated in a cream using, for example, an oil-in-water emulsion base. Such transdermal formulations are well known in the art and typically contain additional ingredients to enhance the skin penetration and stability of the active ingredient or formulation. All such known transdermal formulations and ingredients are included within the scope of this invention.

[0397] The nanoparticles described in this article can also be administered via transdermal devices. Therefore, transdermal drug delivery can be achieved using reservoir-type, porous membrane-type, or solid matrix patches.

[0398] The above-described components for use in oral, injectable, or topical compositions are merely representative. Other materials and processing techniques are described in Part 8 of Remington's Pharmaceutical Sciences, 17th edition, 1985, Mack Publishing Company, Easton, Pennsylvania, which is incorporated herein by reference.

[0399] For injection, the nanoparticles described herein can be provided in injectable saline solutions, as well as in the form of injectable liposome solutions or sustained-release polymer systems.

[0400] The nanoparticles described in this article can also be administered in a sustained-release form or from a sustained-release drug delivery system. Descriptions of representative sustained-release materials can be found at Remington's Pharmaceutical Sciences.

[0401] method

[0402] This article provides methods for treating subjects who are susceptible to or have immune-related diseases and conditions, including, for example, immune paralysis in sepsis and infection, cell proliferation disorders (e.g., cancer), and other diseases and conditions caused by defective trained immunity.

[0403] In embodiments, this disclosure provides a method for treating cell proliferation disorders, comprising administering a therapeutically effective amount of a nanobiocomposition to a subject in need, said nanobiocomposition comprising high-density lipoprotein (HDL)-derived nanoparticles containing a compound of this disclosure (e.g., a compound of Formula I). ​​In embodiments, the compounds and compositions provided herein can be used to treat cancer by inducing immune training.

[0404] In the implementation plan, the cell proliferation disorder is defined as cancer. Specifically, the cancer is one or more of the following: advanced malignant tumors, amyloidosis, neuroblastoma, meningioma, hemangiopericytoma, multiple brain metastases, glioblastoma multiforme, glioblastoma, brainstem glioma, malignant brain tumors with poor prognosis, malignant glioma, recurrent malignant glioma, anaplastic astrocytoma, anaplastic oligodendroglioma, neuroendocrine tumors, rectal adenocarcinoma, Dukes C&D colorectal cancer, unresectable colorectal cancer, metastatic hepatocellular carcinoma, Kaposi's sarcoma, karyotype acute myeloid 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, malignant melanoma, malignant mesothelioma, malignant pleural effusion mesothelioma syndrome, peritoneal cancer, papillary serous carcinoma, gynecologic sarcoma, soft tissue sarcoma, scleroderma, cutaneous vasculitis, Langerhans cell group Histiocytosis, leiomyosarcoma, progressive ossifying fibrous dysplasia, hormone-resistant prostate cancer, resected high-risk soft tissue sarcoma, unresectable hepatocellular carcinoma, Waldenström 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 carcinoma, follicular thyroid carcinoma, medullary thyroid carcinoma, and leiomyosarcoma. In the implementation plan, cancers are selected from the group consisting of: bladder cancer, vascular cancer, bone cancer, brain cancer, breast cancer, cervical cancer, thoracic cancer, colon cancer, endometrial cancer, esophageal cancer, eye cancer, head cancer, kidney cancer, liver cancer, lymph node cancer, lung cancer, oral cancer, neck cancer, ovarian cancer, pancreatic cancer, prostate cancer, rectal cancer, colorectal cancer, skin cancer, gastric cancer, testicular cancer, laryngeal cancer, thyroid cancer, urothelial carcinoma, and uterine cancer. In the implementation plan, the cancer is selected from the group consisting of: breast cancer, prostate cancer, melanoma, colorectal cancer, lung cancer, pancreatic cancer, and glioblastoma. In the implementation plan, the cancer is metastatic. In the implementation plan, the cancer is refractory or resistant to chemotherapy or radiotherapy; in particular, resistant to thalidomide.

[0405] In the implementation plan, cancers are selected from the group consisting of: bladder cancer, vascular cancer, bone cancer, brain cancer, breast cancer, cervical cancer, chest cancer, colon cancer, endometrial cancer, esophageal cancer, eye cancer, head cancer, kidney cancer, liver cancer, lymph node cancer, lung cancer, oral cancer, neck cancer, ovarian cancer, pancreatic cancer, prostate cancer, rectal cancer, colorectal cancer, skin cancer, stomach cancer, testicular cancer, laryngeal cancer, thyroid cancer, urothelial carcinoma, and uterine cancer.

[0406] In the implementation plan, the cancers selected are those consisting of breast cancer, prostate cancer, melanoma, colorectal cancer, lung cancer, pancreatic cancer, and glioblastoma.

[0407] In one embodiment, this disclosure provides a method for treating sepsis, comprising administering a therapeutically effective amount of the nanobiocomposition of this disclosure to a subject in need. In another embodiment, the patient suffers from sepsis associated with a bacterial, viral, or fungal infection of the lungs, abdomen, kidneys, or bloodstream.

[0408] The compounds disclosed herein and their carriers disclosed herein can be used to enhance immune responses. Therefore, methods for inducing immune responses are disclosed herein, comprising administering an immunogenic composition to a subject, wherein said composition optionally comprises (i) at least one antigen and (ii) a compound disclosed herein in a nanoparticle carrier such as HDL-derived nanoparticles or liposomes.

[0409] While neoantigens from subjects with cancer can be used, antigens are typically derived from pathogens. Illustrative pathogen antigens can originate from viruses, bacteria, parasites, or yeast. In some respects, antigens can be secreted by pathogens; for example, exotoxins or endotoxins.

[0410] Exemplary viruses include adenovirus, adeno-associated virus (AAV), chikungunya virus, dengue virus, influenza virus, Ebola virus, Epstein-Barr virus, Hantavirus, hepatitis viruses (e.g., hepatitis A, hepatitis B, hepatitis C, hepatitis D, hepatitis E), CMV, HPV (e.g., one or more of HPV1-18), coronaviruses (e.g., SARS, MERS, COVID-19), poliovirus, rabies virus, and Zika virus. Exemplary bacteria include Vibrio cholerae, Escherichia coli, Salmonella spp., Neisseria gonorrhoeae, Neisseria meningitidis, Streptococcus pyogenes, Mycobacterium tuberculosis, Legionella pneumophila, Brucella, and Listeria monocytogenes.

[0411] Antigens can be, for example, polypeptides comprising glycosylated peptides or carbohydrates. In some aspects, the immunogenic composition may contain nucleic acids encoding antigens, typically polypeptides transcribed and / or translated from said nucleic acids. Nucleic acids can be DNA or RNA, or derivatives of DNA or RNA. Common RNA derivatives include covalent modifications to the molecule to enhance stability and / or expression. In some aspects, the nucleic acid encoding the polypeptide may be contained within plasmids or viral vectors such as adenovirus vectors, adeno-associated virus vectors, baculovirus vectors, and lentivirus vectors.

[0412] In some respects, administration can be prophylactic; for example, to vaccinate a subject before exposure to a pathogen. In other respects, administration can be therapeutic; for example, to induce an immune response against a tumor carrying a neoantigen in a subject with cancer. In an embodiment, the nanobiocomposite is administered to a patient in a treatment regimen comprising two or more doses to generate drug accumulation in myeloid cells, myeloid progenitor cells, and hematopoietic stem cells in the bone marrow, blood, and / or spleen.

[0413] In the implementation plan, the nanobiocomposition is administered intravenously or intra-arterially.

[0414] Injection dose levels range from about 0.1 mg / kg / hour to at least 10 mg / kg / hour, all lasting from about 1 to about 120 hours, and especially 24 to 96 hours. A preloading bolus of about 0.1 mg / kg to more than 10 mg / kg may also be given to achieve adequate steady-state levels. For human patients weighing 40 to 80 kg, the maximum total dose is expected not to exceed about 2 g / day.

[0415] The oral dose levels range from about 0.01 to about 20 mg / kg for the compounds of the present invention, including all ranges and values ​​therein. For example, the dose levels range from about 0.1 to about 10 mg / kg or from about 1 to about 5 mg / kg.

[0416] Transdermal doses are typically chosen to provide similar or lower blood levels as achieved with injectable doses. Suitable administration methods for mucosal sites are also envisioned herein, including, without limitation, anal swabs, enemas, nasal sprays, and nebulized or vaporized compounds and / or compositions for delivery to the pulmonary mucosa. Those skilled in the art will select an appropriate delivery modality based on a wide range of parameters, including the organ or tissue site most severely affected by the disease or condition in a patient.

[0417] The compounds of the present invention can be administered as single active agents, or they can be administered in combination with one or more additional agents, said additional agents including other compounds exhibiting the same or similar therapeutic activity and identified as safe and effective for such combination administration. In embodiments, the additional agents are inhibitors of checkpoint proteins. In embodiments, the methods provided herein further include co-administering a cancer drug with a nanobiocomposite as a combination therapy.

[0418] The compounds or compositions described herein may be provided in the kit. In some embodiments, the kit includes (a) the compounds described herein or compositions comprising the compounds described herein (wherein, for example, the compounds may be NOD2 regulators described herein), and optionally (b) informational materials. The informational materials may be descriptive, instructional, marketing, or other materials relating to the methods described herein and / or the use of the compounds or compositions described herein in the methods described herein. In embodiments, the informational materials may include information regarding the production of the compounds. In embodiments, the informational materials relate to methods of administering the compounds. In embodiments, the informational materials may include instructions on administering the compounds or compositions described herein in a suitable manner, such as at a suitable dose, dosage form, or route of administration (e.g., the dose, dosage form, or route of administration described herein) to perform the methods described herein. In embodiments, the informational materials may include instructions on administering the compounds described herein to suitable subjects (e.g., persons, such as those with or at risk of having the conditions described herein).

[0419] The kit may include one or more containers for a composition containing the compounds or compositions described herein. In embodiments, the kit includes separate containers, separators, or compartments for the composition and 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 pouch. In embodiments, the separate components of the kit are contained in a single, unseparated container. For example, the composition is contained in a bottle, vial, or syringe containing informational material already labeled. In embodiments, the kit includes multiple (e.g., a pack) individual containers, each containing one or more unit dosage forms (e.g., dosage forms described herein) of the compounds or compositions described herein. For example, the kit includes multiple syringes, ampoules, foil pouches, or blister packs, each containing a single unit dose of the compound described herein. The kit containers may be airtight, waterproof (e.g., impermeable to changes in moisture or evaporation), and / or opaque.

[0420] Furthermore, a process for manufacturing the nanobiocompositions of this disclosure is provided herein, the process comprising:

[0421] a) Formation of a lipid membrane comprising: i) compounds 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

[0422] b) Dissolve the lipid membrane in a solvent to form a lipid solution; and contact the lipid solution with apoA-I or a peptide mimic of apoA-I under conditions that effectively form HDL-derived nanoparticles containing compounds of the present disclosure.

[0423] In the embodiments, nanobiocompositions prepared according to the methods described herein are provided.

[0424] Example

[0425] The therapeutic agents and nanoparticles containing them described herein can be prepared by those skilled in the art of organic synthesis from known or commercially available starting materials and reagents.

[0426] Materials and methods

[0427] Unless otherwise specified, all chemicals were purchased from commercial sources and were ready for use without further purification. N-methylmorpholine was redistilled, collecting the fraction from 110°C to 112°C. Cholesterol azidoacetate was synthesized according to known procedures (RSCAdv. 2015, 5, 12094), as was 1-azidooctadecane. Dry solvents were obtained using the MBRAUN solvent purification system (MB-SPS). Toluene was purified before use. Molecular sieve drying. Glassware used in the reaction under argon atmosphere was dried with a hot air gun before use. Thin-layer chromatography (TLC) was performed using Merck 60-F254 silica gel plates and visualized by UV light at 254 nm, permanganate staining, and / or cerium molybdate (CeMo) staining. Biotage was used. Silica, Büchi FlashPure ID, or Büchi FlashPure ID C18 columns were used for normal-phase and reversed-phase automated column chromatography analysis on Biotage Isolera One or Grace Reveleris X2 rapid chromatography systems. The elution gradient was specified in column volume (CV). Unstable THF was used for the water / THF gradient.

[0428] NMR spectra were recorded on a Bruker 400MHz Ultrashield spectrometer (400MHz for 1H NMR). The deuterated solvent used was specified in each case. Chemical shifts (δ) are expressed in ppm and refer to the residual peak of the solvent. Peak multiplicity is abbreviated as s: singlet; d: doublet; t: triplet; dt: double triplet; ddt: double double triplet; td: triple doublet; tt: triple triplet; q: quartet; ABq: AB quartet; dq: double quartet; qd: quadruple doublet; sept: septet; m: multiplet; bs: broad singlet. Matrix-assisted laser desorption / ionization time-of-flight (MALDI-TOF) mass spectrometry was performed 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]malonitrile (DCTB) as the matrix. Gas chromatography-mass spectrometry (GC-MS) measurements were performed on a Shimadzu GC-17A gas chromatograph equipped with a Shimadzu AOC-20i autosampler, a Shimadzu GCMS-QP5000 gas chromatograph-mass spectrometer, and a Phenomenex Zebron ZB-35 column (l = 30 m, ID = 0.25 mm, film thickness = 0.25 μm). High-performance liquid chromatography-mass spectrometry (HPLC-ESI-MS) with a water / acetonitrile gradient was performed using a Shimadzu setup equipped with a 2x LC-20AD pump, a DGU-20A3 degasser, a SIL-20AC autosampler, an SPD-M20APDA, and a Thermo Scientific LCQfleet MS. Column: Phenomenex Kinetex 5μm EVO C18 LC (50 × 2.1 mm). Gradient: Water / MeCN (+0.1% formic acid), from 5% MeCN to 100% MeCN, 0.300 mL / min. Electrospray ionization (ESI) was used to generate the charge for MS detection. HPLC-MS and HPLC-ELSD experiments using water / THF or water / MeOH gradients were performed on a Shimadzu Nexera-i LC-2040C 3D Plus with a Shimadzu LCMS-8045. Column: Alltech Alltima C18 (150 × 3.2 mm; 5 μm; no. 88383). Gradient: Water / THF (+0.1% TFA) or water / MeOH (+0.1% TFA), 0.400 mL / min. This HPLC setup was also used in combination with ELSD (evaporative light scattering detection).

[0429] Optionally, HPLC-MS (SIM) and HPLC-ELSD were performed on a Phenomenex Kinetex 5 μm EVO C18100A LC column (50 × 2.1 mm) using a gradient from eluent A to eluent B, wherein A = 20 mM NH4HCO2 in H2O with 0.1 v / v% formic acid, and B = 2-propanol / MeCN / H2O 85:15:5, also with 20 mM NH4HCO2 and 0.1 v / v% formic acid.

[0430] abbreviation

[0431] HPLC = High Performance Liquid Chromatography; ELSD = Evaporative Light Scattering Detection; ESI-MS = Electrospray Ionization Mass Spectrometry; SIM = Selected Ion Mode; NMR = Nuclear Magnetic Resonance.

[0432] MDP-[53678-77-6]

[0433] MDP = muramyl dipeptide muramyl (or N-acetyl muramyl-L-alanyl-D-isoglutamine) CAS [53678-77-6] prepared according to standard peptide synthesis or purchased from commercial sources.

[0434] NHS = N-hydroxysuccinimide; DiC or DIC = N,N′-diisopropylcarbodiimide; EDC = N-(3-dimethylaminopropyl)-N′-ethylcarbodiimide (using hydrochloride); PyBOP = (benzotriazol-1-yloxytripyrrolidinylphosphonium hexafluorophosphate); SPPS = solid-phase peptide synthesis.

[0435] TEA = Triethylamine; THF = Tetrahydrofuran; MeOH = Methanol; DMF = Dimethylformamide; FA = Formic acid; TFA = Trifluoroacetic acid.

[0436] Constructing blocks

[0437] DSPE-azidoacetic acid ester (DSPE-CO-CH2-N3)

[0438]

[0439] (2R)-3-(((2-aminoethoxy)(hydroxy)phosphoryl)oxy)propane-1,2-dimethyldistearate (260 mg, 0.35 mmol), 2,3,5,6-tetrafluorophenyl 2-azidoacetic acid (prepared according to DJ Vugts et al., Bioconjugate Chem. 2011, 22, 2072-2081; 87 mg, 0.35 mmol, 1 eq), and N,N-diisopropylethylamine (184 μL, 1.1 mmol, 3 eq) 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 with MgSO4, filtering, and removing the solvent under vacuum, the compound was purified by column chromatography (fast SiO2) using an elution gradient of 5% to 40% MeOH in chloroform. This yielded pure DSPE-azidoacetate (244 mg, 0.29 mmol, 84%) as a white solid. 1 H-NMR (400MHz, CDCl3 / CD3OD 9:1): δ=5.23(dt,J=9.0,4.6Hz,1H),4.35(dd,J=12.0,3.7Hz,1H),4.22-3.99(m,5H),3.9 5(s,2H),3.53(t,J=5.1Hz,2H),2.33(q,J=7.6Hz,4H),1.61(td,J=7.4,4.2Hz,4H),1.48-

[0440] 1.16(m,56H),0.88(t,J=6.7Hz,6H). 13 C-NMR (101MHz, CDCl3): δ=173.7,173.4,168.3,69.7,69.6,66.1,66.0,65.2,62.1,52.5,40.01,39.95,34.3,34.2,3 4.1,32.1,29.9,29.80,29.7,29.62,29.59,29.50,29.47,29.46,29.4,29.30,29.26,25.00,24.97,24.9,22.8,14.2. 31 P-NMR (162MHz, CDCl3): δ=-0.48.

[0441] MALDI-TOF MS:C 43 H 83 The calculated m / z value for N4O9P is 830.59; the observed value is [M+Na]. + 853.62, [M-H+2Na] +875.58.

[0442] MDP-propargyl

[0443]

[0444] Fill a 50 mL round-bottom flask with MDP (0.113 g, 0.23 mmol, 1.00 eq). Dissolve the substance in dry DMF (~1.5 mL, 0.15 M) and purge the flask with argon.

[0445] EDC·HCl (0.066 g, 0.34 mmol, 1.50 eq) and N,N-diisopropylethylamine (0.050 g, 0.068 mL, 0.39 mmol, 1.70 eq) and 4-(N,N-dimethylamino)pyridine (0.0028 g, 0.023 mmol, 0.10 eq) were added, and the resulting clear solution was stirred at room temperature for 5 min. Next, propargyl-1-amine (0.018 g, 0.021 mL, 0.32 mmol, 1.40 eq) was added via syringe. Stirring continued at room temperature. After a reaction time of 21 h, complete conversion of the MDP starting material was confirmed by LC-MS (water / MeOH). The reaction mixture was concentrated under vacuum to give a crude product resembling a yellow glass. The substance was purified twice by automated column chromatography (reversed-phase (C18); product: C18-silica 1:100; detection: 200-400 nm), eluting with water / MeOH 90 / 10-82 / 18. The pure fraction was lyophilized to give the pure product (0.050 g, 41%) as a white solid.

[0446] 1 H NMR (400MHz, MeOD) δ5.16(d,J=3.4Hz,1H),4.42-4.23(m,3H),3.95(t,J=2.3Hz,2H),3.94-3.57(m,5H),3.52-3.39(m Hz,1H),2.58(t,J=2.6Hz,1H),2.33-2.26(m,2H),2.25-2.13(m,1H),2.00-1.85(m,4H),1.45-1.33ppm(m,6H). 13C NMR (100MHz, 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. Measured value: m / z = 512.08 [M-H2O+H] + 552.33 [M+Na] + (Positive mode); 325.17[M-murayl]- (Negative mode).

[0447] MDP(Bn)

[0448]

[0449] MDP(Bn) was synthesized in a 100 mL glass reaction vessel with a glass frosted filter bottom using the standard SPPS method. Adequate stirring of the reaction mixture was ensured by applying a constant argon flow through the glass frosted filter, while excess reagents and wash solutions were removed by vacuum filtration. The crude MDP(Bn) was purified twice by automated column chromatography (reversed-phase (C18); product: C18-silica 1:200; detection: 200–400 nm), eluting with water / MeCN + 0.1% formic acid 90 / 10–82 / 18. The pure fraction was lyophilized to give the pure product (0.309 g, 67%) as a fluffy white substance.

[0450] 1H NMR (400MHz, DMF-d7) δ8.18(d,J=8.6Hz,1H),8.15(d,J=8.6Hz,1H),7.75(d,J=6.5Hz,1H),7.53-7.28(m,6H),7. 11-7.03(m,1H),4.86(d,J=3.5Hz,1H),4.76(d,J=12.3Hz,1H),4.72-4.56(m,1H),4.51(d,J=12.3Hz,1H),4.47-4 .32(m,3H),4.01(ddd,J=10.7,8.4,3.5Hz,1H),3.83(dd,J=11.6,2.2Hz,1H),3.78-3.61(m,3H),3.60-3.40(m,1 H), 2.39 (t, J = 7.8Hz, 2H), 2.24-2.12 (m, 1H), 1.97-1.83 (m, 4H), 1.40 (d, J = 7.0Hz, 3H), 1.34ppm (d, J = 6.7Hz, 3H). 13 C NMR (100MHz, 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] + .

[0451] MDP(Bn)-propargyl

[0452]

[0453] Under an argon atmosphere, MDP(Bn) (0.110 g, 0.19 mmol, 1.00 eq) was added to a 5 mL round-bottom flask. This substance was dissolved in dry DMF (0.5 mL). PyBOP (0.127 g, 0.25 mmol, 1.30 eq) and N,N-diisopropylethylamine (0.049 g, 0.066 mL, 0.38 mmol, 2.00 eq) were added, yielding a clear, colorless solution. The mixture was stirred at room temperature for 5 minutes. Next, propargyl-1-amine (0.021 g, 0.024 mL, 0.38 mmol, 2.00 eq) was added, and the resulting pale yellow mixture was stirred at room temperature. After a 2-hour reaction time, the reaction mixture was concentrated under vacuum to give a crude product as a beige, viscous solid. The substance was purified by automated column chromatography (reversed-phase (C18); product: C18-silica 1:100; detection: 200-400 nm), eluted with water / MeCN 90 / 10-80 / 20. The pure fraction was lyophilized to give the pure product (0.096 g, 82%) as a white solid.

[0454] 1 H NMR (400MHz, DMF-d7) δ8.25(t,J=5.5Hz,1H),8.20-8.15(m,2H),7.75(d,J=6.6Hz,1H),7.51-7.28(m,6H),7.08-7.03(m, 1H),5.47(d,J=6.3Hz,1H),4.86(d,J=3.5Hz,1H),4.76(d,J=12.3Hz,1H),4.63(t,J=6.0Hz,1H),4.51(d,J=12.4Hz,1H), 4.47-4.27(m,3H),4.05-3.95(m,3H),3.83(ddd,J=11.5,5.7,2.2Hz,1H),3.75-3.61(m,3H),3.54-3.46(m,1H),3.04(t, J=2.5Hz,1H),2.32-2.26(m,2H),2.21-2.11(m,1H),1.95-1.83(m,4H),1.39(d,J=7.0Hz,3H),1.34ppm(d,J=6.8Hz,3H). 13C NMR (100MHz, DMF-d7) δ 173.79, 173.59, 172.74, 171.93, 170.19, 138.47, 128.58, 127.93, 127.80, 97.05, 81.35, 80.16, 77.36, 73.94, 72.14, 70.63, 68.62, 61.76, 53.65, 52.94, 49.38, 35.63, 32.21, 30.47, 28.31, 28.25, 22.66, 19.01, 18.06 ppm. HPLC-MS (water / MeCN): t(product) = 3.35 min. Measured value: m / z = 620.17 [M+H] + .

[0455] MTP-b on resin

[0456]

[0457] MTP-b on resin was synthesized using the standard SPPS method in a 100 mL glass reaction vessel with a glass frosted filter bottom. Adequate stirring of the reaction mixture was ensured by applying a constant argon flow through the glass frosted filter, while excess reagents and wash solutions were removed by vacuum filtration. After a final coupling wash, the resin was washed again with DCM (2 × 20 mL) and dried under an argon flow. The material was stored at -20 °C. The sample was lysed from the resin using TFA / TIPS / water 95 / 2.5 / 2.5 (0.1 mL, 10 min) and examined by HPLC-MS (water / MeCN). HPLC-MS (water / MeCN): t(product) = 3.65 min. Analytical value: m / z = 874.42 [M+H] + (Positive mode); m / z = 918.08

[0458] [M+HCOO] - (Negative mode)

[0459] MTP-b-N3

[0460]

[0461] MTP-b (429 mg, approximately 0.15 mmol MTP-b) was added to the resin in a 20 mL PE syringe containing PE frit, and the resin was allowed to swell in DMF (12 mL) for 30 min. The resin was treated twice with a 2% hydrazine hydrate solution in DMF (12 mL), 15 min each time. 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 eq), and N,N-diisopropylethylamine (0.34 mL, 1.9 mmol, 12 eq) in DMF (12 mL) was added to the resin, and the glass beads were stirred at room temperature for 24 h (with occasional release of slight overpressure). After filtration, the resin was washed with DMF (5 × 12 mL) for 1 min and then with dichloromethane (4 × 10 mL) for 1 min. The resin was then lysed for 2 h in a TFA / TIPS / H2O 95:2.5:2.5 (4 mL) solution. After filtration, the resin was washed with TFA (4 mL) for 5 min. The combined TFA filtrate was concentrated under vacuum (keeping the temperature as low as possible to avoid TFA-ester formation). An impure compound was obtained by automated column chromatography (reversed phase (C18); detection: λ = 200 nm) using an elution gradient of 5% to 60% MeCN (all containing 0.1% TFA) in H2O. This was further purified by RP-HPLC using an elution gradient of 26% to 35% MeCN (all containing 0.1% TFA) in H2O, and the product was obtained as a pure white, fluffy solid (37.5 mg, 51 μmol, 34%) after lyophilization. 1H-NMR (400MHz, DMF-d7 / D2O9:1): δ = 8.45 (t, J = 9.1Hz, 2H), 8.26 (d, J = 7.9Hz, 1H), 7.99 (d, J = 6.5Hz, 1H), 7.81 (d, J = 2.8Hz, 2H), 7.64-7.45 (m ,5H),7.31(d,J=17.2Hz,2H),5.82(d,J=6.3Hz,1H),5.03(d,J=3.5Hz,1H),4.93(d,J=12.4Hz,1H),4.68(d,J=12.4Hz,1H),4.63-4.46(m,4H ),4.19(dd,J=10.7,3.6Hz,1H),4.00(dd,J=6.8,6.3Hz,1H),3.92-3.79(m,3H),3.66(t,J=9.0Hz,1H),3.52(t,J=6.8Hz,2H),2.53(t,J=7.5 Hz,2H),2.37(dtd,J=16.5,7.9,4.3Hz,1H),2.10(s,3H),2.09-1.93(m,2H),1.88-1.55(m,5H),1.58(d,J=7.1Hz,3H),1.52(d,J=6.7Hz,3H). 13 C-NMR (100MHz, DMF-d7 / D2O 9:1):δ=174.9,174.24,174.17,174.00,173.93,173.14,173.06,172.9,17 2.8,170.90,170.8,138.2,128.6,127.9,127.8,96.8,80.0,77.3,73.7,70 .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: C 32 H 49 N9O 11 The calculated m / z value is 735.36; the observed value is [M+H]. + 736.25, [M+Na] + 758.42.

[0462] DSG 4-Nitrophenyl Carbonate

[0463]

[0464] A 25 mL round-bottom flask was filled with a solution of commercially available [(2S)-3-hydroxy-2-octadecanoyloxypropyl]octadecanoate (0.601 g, 0.96 mmol, 1.00 eq) in chloroform (6.5 mL, ~0.15 M). Pyridine (0.122 g, 0.125 mL, 1.54 mmol, 1.60 eq) was added, and the resulting clear solution was cooled in ice water. Next, solid 4-nitrobenzene chloroformate (0.252 g, 1.25 mmol, 1.30 eq) was added in small portions. The pale yellow reaction mixture was stirred overnight at room temperature. Complete conversion of the alcohol was achieved by... 1 The reaction mixture was then precipitated in MeOH (100 mL) and collected by filtration through a glass filter. The mixture was washed with MeOH (30 mL total) and Et₂O (10 mL total) and dried in a vacuum oven at 30 °C. The product was obtained as a white solid (0.713 g, 94%).

[0465] 1 H NMR (400MHz, CDCl3) δ8.29(d,J=9.1Hz,2H),7.39(d,J=9.1Hz,2H),5.38(p,J=5.2Hz,1H),4.50(dd,J=11.7,3.9Hz,1H),4.41-4.32( m,2H),4.22(dd,J=12.0,5.6Hz,1H),2.35(dt,J=9.8,7.5Hz,4H),1.68-1.58(m,4H),1.37-1.17(m,56H),0.88ppm(t,J=6.7Hz,6H). 13 C NMR (101MHz, CDCl3) δ173.24,172.92,155.35,152.29,145.56,125.36,121.76,68.33,66.97,61.63,3 4.16,34.03,31.93,29.71,29.68,29.64,29.49,29.37,29.29,29.13,29.07,24.87,22.70,14.12ppm.

[0466] 2,3,5,6-Tetrafluorophenyl stearate

[0467]

[0468] Triethylamine (2.8 mL, 5 eq.) was slowly added to a solution of stearoyl chloride (1.26 g, 4.2 mmol) and tetrafluorophenol (0.72 g, 1.04 eq.) in DCM (10 mL), resulting in the immediate formation of a white precipitate. The heterogeneous reaction mixture was stirred for another 2 hours, diluted with 25 mL of DCM, and 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 (supplemented with a small amount of activated carbon) and evaporated to dryness. The resulting crude substance was redissolved in chloroform, rinsed over a silica plug, and evaporated to dryness again to give 1.2 g (67%) of the desired compound as a white solid. 1 HNMR (400MHz, CDCl3) δ6.98(tt,J=9.9,7.0Hz,1H),2.66(t,J=7.4Hz,2H),1.78(p,J=7.4Hz,2H),1.26(s,28H),0.88(t,J=6.7Hz,3H)ppm. 19 FNMR(376MHz, CDCl3)δ-139.21(ddd,J=24.0,11.8,7.8Hz),-153.03-

[0469] -153.20(m). 13 C NMR (101MHz, CDCl3) δ169.56,147.23(m),144.74(m),141.94,141.86(m),139.41(m),129.78(m),103. 01(t),33.42,31.92,29.69,29.65,29.61,29.54,29.39,29.36,29.14,28.85,24.78,22.68,14.09ppm.

[0470] Synthesis method

[0471] First method. The compounds of the present invention can be prepared using starting reactants MDP or MDP(Bn) – see above. These molecules have functional carboxylic acid groups derived from glutamic acid (Glu) constitutive blocks. The COOH group enables conjugation with 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 cholesterol-derived moieties. Particularly useful constitutive blocks are PE-phospholipids such as DSPE ([1069-79-0]) or DOPE ([4004-05-1]); these molecules are already amine-functional. Mixed acyl PE-phospholipids can also be useful (e.g., 16:0-18:1 PE or 18:0-18:1 PE or 18:0-16:0 PE). Another useful constitutive block is cholesterol. Another useful constituent block is a diglyceride, such as 1,2-bis(octadecanoyl-sn-glycerol) (18:0DG[51063-97-9]) or 1,2-dioleoyl-sn-glycerol (18:1DG[24529-88-2]). Mixed acylglycerols (e.g., 16:0-18:1DG or 18:0-18:1DG or 18:0-16:0DG) can also be used. PE-phospholipids and diglycerides have two lipophilic chains, and such constituent blocks are preferred for this method.

[0472] The first method is illustrated in Examples 10 to 19.

[0473] The second method. In a particularly suitable alternative modular approach, a copper-catalyzed azide-alkyne cycloaddition ("click reaction") is used to link MDP (or MTP) reactants to lipophilic reactants. Here, MDP, MDP(Bn), MTP, or MTP(Bn) with azide (-N3) or alkyne (-C≡CH) functional groups are used to form blocks. Non-limiting examples of such molecules are MDP-propynyl, MDP(Bn)-propynyl, or MTP-b-N3—see above. In the copper-catalyzed click reaction, these molecules can be coupled to alkyne-functionalized or azide-functionalized molecules containing lipophilic groups. Using the click reaction, targeted, modular, and stable azide-functionalized or alkyne-functionalized intermediates can be readily and modularly prepared. This simplifies the isolation and storage of the intermediates. Furthermore, copper-catalyzed click cycloaddition can and preferably takes place in an aqueous environment (e.g., THF / water or tBuOH / water) or in an aqueous two-phase liquid / liquid solvent combination (e.g., dichloromethane / water). In these reaction media, both the hydrophilic MDP (or MTP) reactant (with or without a Bn group) and the lipophilic reactant are readily soluble, significantly improving the ease of conjugation and reaction yield. In this click method, the lipophilic group contained in the azide or alkyne reactant can be a C14, C16, or C18 moiety, such as those derived from stearic acid, palmitic acid, myristic acid, oleic acid, palmitoleic acid, myristone acid, stearyl alcohol or amine, palmitol or amine, myristol or amine, oleyl alcohol or amine, palmitoleyl alcohol or amine, myristenol or amine; or a sterol moiety, such as those derived from cholesterol. Saturated linear lipophilic moieties are preferred, as are those derived from cholesterol. Particularly useful constituent blocks are PE-phospholipids, mixed acyl PE-phospholipids, diglycerides (DG), or mixed acyl diglycerides (containing C14, C16, and / or C18 moieties), and cholesterol. Compositions containing two lipophilic chains or lipophilic azides or alkynes containing cholesterol groups are preferred.

[0474] The second method is shown in Examples 1 to 9.

[0475] Note that both methods allow for the introduction of additional amino acid units linked to the glutamate unit of the MDP or MDP(Bn). Suitable amino acid units are those derived from L-lysine or L-alanine. With the addition of an additional amino acid unit, the MDP (muramylated dipeptide) portion is converted to the MTP (muramylated tripeptide) portion, with or without the Bn group. Examples are shown in Examples 9-10, 13-15, and 17-18.

[0476] Example 1. Synthesis of MDP-C18 [click] (1)

[0477]

[0478] Molecular weight: 825 Daltons. CLogP = 4.15.

[0479] This synthesis demonstrates the general conditions for Cu click reactions.

[0480] MDP-propargyl (0.011 g, 0.02 mmol, 1.00 eq) was added to a 5 mL vial. L-Ascorbic acid (0.4 M aqueous solution, 104 μL, 41.5 μmol ascorbic acid, 2.00 eq) was then added to the slightly opaque solution. A solution of 1-azidooctadecane (0.012 g, 0.04 mmol, 2.00 eq) in DCM (0.8 mL) was added, followed by an aqueous solution of copper(II) sulfate pentahydrate (0.2 M, 104 μL, 20.8 μmol Cu, 1.00 eq). The bilayer reaction mixture was then stirred at 1400 rpm at room temperature to obtain a pale yellow / green emulsion. After 16 h, the reaction mixture was concentrated under a nitrogen stream to obtain a crude product as a light brown sludge. This substance was absorbed in chloroform / MeOH 4:1 and impregnated on diatomaceous earth (90 mg, ~1:5 loading ratio). Purification was performed by automated column chromatography (product: silica 1:500; detection: 200-400 nm), eluting with chloroform / MeOH / water 90 / 9 / 1-70 / 27 / 3 to give the product as a white solid (0.005 g, 31%).

[0481] 1 H NMR (400MHz, MeOD) δ7.64-7.61(m,1H),5.33(d,J=3.4Hz,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),0.88ppm(t,J=6.8Hz,3H). 13C NMR (100MHz, MeOD) δ175.75,174.66,174.04,173.58,171.91,144.63,122. 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. 49,26.60,22.83,22.79,22.69,19.34,16.95,16.57,14.15ppm. HPLC-MS (water

[0482] / MeCN): t(product) = 5.64 min. Measured value: m / z = 825.33[M+H]+.

[0483] Example 2. Synthesis of MDP-DSPE (2)

[0484]

[0485] Molecular weight: 1361 Daltons. CLogP = 11.56 (uncharged) and 5.78 (negatively charged).

[0486] MDP-propargyl (24.4 mg, 46 μmol) was dissolved in 0.4 M ascorbic acid (0.24 mL, 2 eq) and a solution of DSPE-azidoacetate (38.5 mg, 46 μmol, 1 eq) in dichloromethane (0.5 mL) was added. Under vigorous stirring, 0.2 M CuSO4·5H2O (0.24 mL, 1 eq) was added, and the two-phase system was stirred vigorously at room temperature for 19 h. The solvent was removed under vacuum, and the green solid was subjected to column chromatography (fast SiO2) using an elution gradient of 20% to 50% MeOH in chloroform, ending in (45% MeOH + 5% H2O) in chloroform (a large amount of the compound was eluted only after the addition of H2O). This yielded an impure product, which was further analyzed by automated column chromatography (reversed-phase (C2O)). 18 Product: C 18 -Silica 1:200; detection: λ=210nm) was further purified using an elution gradient of 25% to 70% THF in H2O. After lyophilization, the pure product (19.5 mg, 14 μmol, 31%) was given as a white, fluffy solid.

[0487] 1H NMR(400MHz, CDCl3+MeOD)δ7.93(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=12.1,6.8Hz,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 peaks between 4.9 ppm and 4.6 ppm are not visible due to overlap with the H₂O peak. MALDI-TOF MS: C 65 H 118 N9O 19 The calculated m / z value of P is 1359.83; the observed value is [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).

[0488] Example 3. Synthesis of MDP-chol (3)

[0489]

[0490] Molecular weight: 999 Daltons. CLogP = 5.04.

[0491] MDP-propargyl (25 mg, 47 μmol) was dissolved in 0.4 M ascorbic acid (0.24 mL, 2 eq), and a solution of cholesterol azide acetate (26.6 mg, 57 μmol, 1.2 eq) in dichloromethane (0.5 mL) was added. Under vigorous stirring, 0.2 M CuSO4·5H2O (0.24 mL, 1 eq) was added, and the two-phase system was stirred vigorously at room temperature for 17 h. H2O / saline 1:1 (50 mL) was added, and the blue aqueous layer was extracted with chloroform / MeOH 2:1 (5 × 20 mL). The combined organic layers were dried over Na2SO4, filtered, and the solvent was removed under vacuum. The resulting colorless solid was repeatedly purified by column chromatography (fast SiO2) using an elution gradient of 6% to 20% MeOH in chloroform. After lyophilization from THF / H2O, a pure product (24.4 mg, 24 μmol, 52%) was obtained as a white, fluffy solid. 1 H-NMR (400MHz, THF-d8 / D2O95:5): δ=7.81(s,1H),5.28(d,J=4.9Hz,1H),5.16(s,2H),5.10(d,J=3. 4Hz,1H),4.59-4.46(m,1H),4.46-4.16(m,5H),3.75-3.17(m,6H),2.26(d,J=8.2Hz,2H),2.20(t,J =7.5Hz,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,5 H),1.13-0.97(m,4H),0.94(s,3H),0.84(d,J=6.5Hz,3H),0.77(dd,J=6.6,1.4Hz,8H),0.61(s,3H). 13 C-NMR (100MHz, 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 .7,76.7,75.4,72.0,70.2,61.1,56.8,56.2,54.0,52.6,50.5,50.2,49.5,42.2,3 9.8,39.4,37.7,36.8,36.4,36.1,35.8,34.4,31.9,31.8,29.6,28.1,27.9,27.5, 27.4,22.5,22.2,22.1,21.9,20.9,18.8,18.7,18.2,16.8,13.5,11.3.MALDI-TOF MS:C 51H 82 N8O 12 The calculated m / z value is 998.60; the observed value is [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 optimal solvent combination for NMR characterization. However, the spectra overlapped and were quite complex. Therefore, integration was tentative.

[0492] Example 4. Synthesis of MDP-DSPE2 [click] (4)

[0493]

[0494] Molecular weight: 2192 Daltons.

[0495] Example 5. MDP-Chol2 [Click]

[0496]

[0497] Molecular weight: 1669 Daltons. CLogP = 15.31.

[0498] Example 6. MDP-DSG [Click] (6)

[0499]

[0500] Molecular weight: 1238 Daltons. CLogP = 12.45.

[0501] Example 7. MDP(Bn)-DSPE [Click](7)

[0502]

[0503] Molecular weight: 1451 Daltons. CLogP = 13.85 (uncharged) and 8.06 (negatively charged).

[0504] MDP(Bn)-propargyl (0.028 g, 0.045 mmol, 1.00 eq) and DSPE-azidoacetate (0.039 g, 0.047 mmol, 1.05 eq) were reacted overnight under the general conditions for Cu click reactions. During the reaction, some substances precipitated out, giving a white suspension / emulsion. The reaction mixture was then diluted with chloroform / MeOH 1:1. The resulting clear solution was impregnated onto diatomaceous earth (~200 mg, 1:3 loading ratio). The impregnated crude product was first purified by automated column chromatography (reversed phase (C18); product: C18-silica 1:200; detection: ELSD and UV 200-400 nm), eluting 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 / 1-75 / 22.5 / 2.5. The pure fractions were concentrated under vacuum, dissolved in water / THF 70 / 30, and lyophilized. Thus, a pure product (0.027 g, 41%) was obtained as a white, fluffy solid.

[0505] 1 H NMR (400MHz, CDCl3+MeOD 1:1)δ7.99(d,J=8.2Hz,1H),7.90(s,1H),7.43-7.23(m,5H),5.24(m,1H),5.13(s,2H),4.9 3(d,J=3.5Hz,1H),4.73(d,J=12.0Hz,1H),4.51(d,J=12.0Hz,2H),4.48-4.40(m,2H),4.37-

[0506] 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),1.67-1.5 6(m,4H),1.52-1.19(m,72H),0.89ppm(t,J=6.8Hz,6H).13CNMR(100MHz,MeOD )δ174.97,174.73,173.93,173.66,173.57,173.46,171.92,166.34,137.16,1 28.28,128.09,127.84,124.43,96.38,79.09,76.52,72.55,70.40,69.75,69.28,63.66,63.44,62.56,61.14,53.18,52.60,52.13,49.45,40.52,34.62,34 .12,33.96,31.81,31.73,29.57,29.53,29.43,29.41,29.23,29.21,29.03,29 .00,27.12,24.82,24.77,22.52,22.20,18.70,16.75,13.69ppm.MALDI-TOF: C 72 H 124 N9O 19 P + 2Na + -H + Calculated m / z value: 1494.85[M+2Na-H] + Measured value: 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).

[0507] Example 8. Synthesis of MDP(Bn)-chol[click](8)

[0508]

[0509] Molecular weight: 1089 Daltons. CLogP = 6.83.

[0510] Under general conditions for Cu click reactions, MDP(Bn)-propargyl (0.030 g, 0.048 mmol, 1.00 eq) and cholesterol azide acetate (0.025 g, 0.053 mmol, 1.10 eq) were reacted overnight to give a white emulsion. The reaction mixture was then concentrated under vacuum and impregnated onto diatomaceous earth (150 mg). The impregnated crude product was purified by automated column chromatography (reversed 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 fraction was concentrated in a vacuum to give a pure product as a white solid (0.028 g, 53%).

[0511] 1 H NMR (400MHz, CDCl3+MeOD 1:1)δ7.78(s,1H),7.39-7.28(m,5H),5.38(d,J=5.1Hz,1H),5.16(d,J=1.6Hz,2H),4.93(d,J=3.6Hz,1H),4.70(d,J=11.8Hz,2H),4 .56-4.37(m,3H),4.30(dt,J=8.9,4.4Hz,1H),4.27-4.17(m,2H),4.02(dd,J=10.1,3.6Hz,1H),3.93-3.67(m,24H),3.67-3.53(m,3H ),3.40(d,J=3.2Hz,0H),2.44-2.32(m,2H),2.27(td,J=7.1,3.7Hz,2H),2.12(dtd,J=14.9,7.5,4.3Hz,1H),2.07-1.74(m,8H),1.72 -1.42(m,5H),1.38(dd,J=13.0,7.0Hz,7H),1.34-0.94(m,13H),0.92(d,J=6.4Hz,3H),0.87(dd,J=6.6,1.8Hz,6H),0.69ppm(s,3H). 13C NMR (100MHz, CDCl3+MeOD 1:1)δ175.17,174.60,173.71,173.62,171.76,166.21,145.18,139.13,137.21,128.65,128.37,128.25,124 .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 ,27.58,24.39,23.95,22.87,22.80,22.77,22.61,21.16,19.34,18.98,18.80,16.90,11.94ppm.MALDI-TOF: C 58 H 88 N8O 12 +Na + The calculated m / z value is: 1111.64 [M+Na] + Measured value: 1111.65. HPLC-MS (water / THF, gradient: 65-95% THF): t(product) = 2.79 min; m / z = 1089.70 [M+H] + (SIM mode).

[0512] Example 9. Synthesis of MTP-b-C18[invclick](9)

[0513]

[0514] Molecular weight: 1058 Daltons. CLogP = 5.68.

[0515] MTP-b-N3 (26 mg, 35 μmol) and propargyl stearate (11.3 mg, 35 μmol, 1 eq) were suspended in THF (0.36 mL) and 0.4 M ascorbic acid (0.18 mL, 2 eq) were added to give a clear solution. Propargyl stearate was prepared using known procedures. 0.2 M CuSO4·5H2O (0.18 mL, 1 eq) was added under vigorous stirring and the mixture was stirred vigorously at room temperature (initially gelation occurred, but gentle heating gave a yellow solution). After 1 h, HPLC-MS (THF / H2O) showed no presence of the starting compound and the opaque solution was lyophilized. The crude product was adsorbed from chloroform / MeOH 2:1 onto diatomaceous earth and subjected to column chromatography (fast SiO2) using an elution gradient of 10% to 25% MeOH in chloroform. Column chromatography was repeated using a similar gradient, and the product was obtained as a pure white fluffy solid (31.5 mg, 30 μmol, 84%) after lyophilization from THF / H2O. 1 H-NMR (400MHz, THF-d8 / D2O 4:1): δ=7.92(s,1H),7.31(d,J=7.5Hz,2H),7.23(t,J=7.5Hz,2H),7.15(t,J=7.3Hz,1H),5.04(s,2H),4.74(d,J=3. 5Hz,1H),4.63(d,J=12.2Hz,1H),4.40(d,J=12.2Hz,1H),4.31-4.15(m,6H),3.96(dd,J=10.5,3.6Hz,1H),3.66(d,J= 3.2Hz,2H),3.59-3.45(m,3H),2.22(dt,J=15.3,7.7Hz,4H),2.09(tt,J=12.9,6.0Hz,1H),1.87-1.70(m,6H),1.64-1 .54(m,1H),1.47(q,J=7.3Hz,2H),1.32(d,J=7.2Hz,3H),1.28(d,J=6.7Hz,3H),1.18(s,30H),0.78(t,J=6.6Hz,3H). 13C-NMR (100MHz, THF-d8 / D2O 4:1): δ=175.5,174.9,174.8,173.73,173.66,173.2,171.7,142.3,137.8,1 28.1,128.0,127.4,124.3,96.6,80.0,77.3,72.8,69.1,68.8,60.9,57.2,5 3.2,53.1,52.2,49.7,49.5,33.6,31.8,31.5,30.9,29.7,29.54,29.50,29. 4,29.24,29.20,29.0,27.7,22.54,22.49,21.9,18.6,16.9,13.5.MALDI-TOF MS:C 53 H 87 N9O 13 The calculated m / z value is 1057.64; the observed value is [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).

[0516] Example 10. Synthesis of MTP-b-C18(10)

[0517]

[0518] Molecular weight: 976 Daltons. CLogP = 5.31.

[0519] Add MTP-b (137 mg, approximately 0.0493 mmol MTP-b, 1.00 eq) to the resin in a 10 mL PE syringe containing PE glass frit. Swell the resin in DMF (5 mL) for 30 min. Next, treat the resin twice with a 2% hydrazine hydrate solution in DMF (10 mL), 15 min each time. Remove the hydrazine solution and wash the resin with DMF (4 × 5 mL). Next, add a solution of 2,3,5,6-tetrafluorophenyl stearate (0.064 g, 0.15 mmol, 3.00 eq) and 4-methylmorpholine (0.030 g, 0.033 mL, 0.30 mmol, 6.00 eq) in DMF / DCM (1 + 1 mL; ~0.075 M). Stir the glass beads overnight at room temperature. The supernatant was then removed and the resin was washed with DMF / DCM 50 / 50 (4 × 5 mL) and DCM (2 × 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 a separate lysis mixture. The combined filtrates were concentrated under vacuum to give a crude product as a white solid. This substance was impregnated from THF / water solution (95 / 5) onto diatomaceous earth (200 mg, 1:4 loading ratio). The impregnated crude product was purified by automated column chromatography (reversed phase (C18); product: C18-silica 1:250; detection: ELSD) eluting with water / THF 50 / 50-10 / 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 / 10-70 / 30. The purified fractions were concentrated under vacuum to give the product as a white solid (0.016 g, 33%). 1 ¹H NMR (400MHz, 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. Measured value: m / z = 976.33 [M+H] + (Positive mode); 1020.25 [M+HCOO] - (Negative mode)

[0520] Example 11. Synthesis of MDP-C18(11)

[0521]

[0522] Molecular weight: 744 Daltons. CLogP = 4.79.

[0523] MDP (10 mg, 20 μmol), octadecylamine (5.2 mg, 0.95 eq.), NHS (2.4 mg, 1 eq.), and EDC-HCl (7.9 mg, 2 eq.) were stirred in DMF (0.7 mL) at 50 °C for 3 hours. The reaction mixture was then cooled to room temperature and stirred for another 16 hours. The resulting dispersion was heated to 40 °C to redissolve all the precipitated solids and then precipitated with 5 mL of diethyl ether. The collected precipitate was washed twice with diethyl ether, dried, and then suspended in softened water. It was collected by centrifugation, resuspended in softened water, and collected again by centrifugation. The resulting solid was lyophilized to remove all water, yielding 13.8 mg (96%) of the desired compound as a white powder. 1 ¹H NMR (400MHz, DMF-d⁷) δ 8.31 (d, J = 7.7Hz, minor isomer), 8.23 ​​(d, J = 8.0Hz, major isomer), 8.12 (d, J = 7.9Hz, minor isomer), 8.09 (d, J = 7.6Hz, major isomer), 7.96 (d, J = 6.6Hz, minor isomer), 7.90 (d, J = 6.5Hz, major isomer), 7.77 (m, ¹H), 7.47 (m, ¹H), 7.10 (m, minor isomer), 7.01 (m, major isomer), 6.86 (d, J = 6.0Hz, minor isomer), 6.74 (dd, J = 4.1, 1.2 Hz, major isomer), 5.44-5.29 (m, 1H), 5.16 (t, J = 3.7 Hz, major isomer), 4.79 (t, J = 6.1 Hz, minor isomer), 4.60 (dd, J = 8.2, 6.0 Hz, minor isomer), 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. 13CNMR(101MHz,DMF-d7)δ174.25,174.20,173.93,173.89,172.75,172.56,172.07,1 72.02,171.76,170.11,96.92,91.66,82.46,79.55,77.39,77.00,72.97,71.30,70. 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) - ).

[0524] Example 12. Synthesis of MDP-DSPE(12)

[0525]

[0526] Molecular weight: 1223 Daltons. CLogP = 12.96 (uncharged) and 7.18 (negatively charged).

[0527] MDP (14 mg, 29 μmol), NHS (5.6 mg, 1.7 eq.), and DIC (7.3 mg, 2 eq.) were stirred in 0.9 mL of DMF for 2 hours to activate MDP. The resulting mixture was added to a dispersion of DSPE (17 mg, 0.8 eq.) in 2.7 mL of tert-butanol and TEA (9 mg, 3.1 eq.) at 50 °C and stirred at this temperature for 3 hours. 1 / 2 hours. The resulting mixture was evaporated to dryness and the resulting substance 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 substance after lyophilization from water / THF.

[0528] 1H-NMR (400MHz, CDCl3 / CD3OD5:) δ 5.29 (d, major isomer), 5.24 (m, 1H), 4.54 (d, minor isomer), 4.47 (m, HDO-masked), 4.41 (dd, HDO-masked), 4.31 (m, HDO-masked), 4.19 (dd, 1H), 4.00–3.90 (m, mixture of isomers), 3.87–3.75 (m, mixture of isomers) (particles), 3.72 (m, 1H), 3.63 (m, 1H), 3.53-3.30 (m, shaded 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 (162MHz, CDCl3 / CD3OD 5:1) δ0.14(br.m)ppm. 13 C-NMR (101MHz, CDCl3 / CD3OD 5:1)δ174.63,174.00,173.27,173.08,172.92,172.89,171.34,90.19,75.19,71. 09,70.18,69.73,69.65,63.24,62.76,61.86,60.84,53.13,52.04,48.77,48.19,3 9.71, 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), measured: 1220.82 (MH). - ), negative mode. HPLC-ELSD (C18, 65-95% THF / H2O): singlets with shoulders for α and β isomers.

[0529] Example 13. Synthesis of MTP-b-DSG(13)

[0530]

[0531] Molecular weight: 1361 Daltons. CLogP = 15.05.

[0532] Add MTP-b (0.291 g, approximately 0.11 mmol MTP-b, 1.00 eq) to the resin in a 20 mL PE syringe containing PE glass frit. Swell the resin in DMF (5 mL) for 30 min. Next, treat the resin twice with a 2% hydrazine hydrate solution in DMF (5 mL), 15 min each time. Remove the hydrazine solution and wash the resin with DMF (4 × 5 mL). Next, add a solution of DSG 4-nitrobenzene carbonate (0.249 g, 0.32 mmol, 3.00 eq) and N,N-diisopropylethylamine (0.081 g, 0.110 mL, 0.63 mmol, 6.00 eq) in chloroform (4 mL). Stir the glass beads overnight at room temperature. Afterward, remove the bright yellow supernatant and wash the resin 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 injected into ice-cold ether (100 mL) with stirring, resulting in the slow formation of white flocculent material. This lysis and precipitation step was repeated twice. The solid was collected by filtration through a disposable PE filter, yielding a crude product as a white solid. This substance was impregnated from chloroform / MeOH (1:2) solution onto diatomaceous earth (250 mg, 1:2.5 loading ratio). The impregnated crude product was purified by automated column chromatography (reversed phase (C18); product: C18-silica 1:150; detection: 200-400 nm), eluting 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. The purified fractions were concentrated under vacuum to give the product as a white solid (0.030 g, 21%).

[0533] 1H NMR (400MHz, CDCl3+MeOD) δ7.39-7.27(m,5H),5.25(p,J=5.3Hz,1H),4.89(d,J=3.5Hz,1H),4.73(d,J=11.9Hz,1H),4.50 (d,J=11.9Hz,1H),4.42-4.31(m,2H),4.31-4.19(m,4H),4.15(dd,J=11.9,6.2Hz,2H),4.10-3.97(m,1H),3.87-3.75(m, 2H),3.72-3.53(m,3H),3.12(t,J=6.9Hz,2H),2.37-2.27(m,6H),2.26-2.13(m,1H),1.93(s,3H),1.92-1.74(m,2H),1.7 1-1.56(m,6H),1.56-1.47(m,2H),1.43(d,7.0Hz,3H),1.40(d,7.0Hz,3H),1.36-1.19(s,56H),0.89ppm(t,J=6.7Hz,6H). 13 C NMR (100MHz, CDCl3+MeOD) δ175.80,174.79,174.30,173.86,173.76,173.44,173.25,171.70,156.58,137.07,12 8.36,128.14,127.95,96.52,79.36,76.83,72.38,69.48,69.43,69.37,62.50,62.29,61.30,53.52,53.24,51.7 1, 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. Measured value: m / z = 1360.9 [M+H] + and 1382.9[M+Na] + (SIM mode).

[0534] Example 14. Synthesis of MTP(Bn)-a-DPPE(14)

[0535]

[0536] Molecular weight: 1328 Daltons. CLogP = 12.88 (uncharged) and 7.09 (negatively charged).

[0537] Constructing a block (CBz)-Ala-DPPE

[0538] N-CBz-protected L-alanine (390 mg, 1.7 mmol) and N-hydroxysuccinimide (222 mg, 1.89 mmol, 1.1 eq) were dissolved in chloroform (6 mL) to give an almost clear solution. N,N′-diisopropylcarbodiimide (DIC; 0.32 mL, 2.0 mmol, 1.2 eq) was added and the mixture was stirred at room temperature for 40 min (after 1 min, the solution became cloudy, and after 25 min, 1H-NMR showed complete conversion). This solution was then added to a 60 °C solution (12 mL; DPPE and triethylamine dissolved under reflux were added at a lower temperature) containing 1,2-dipalmitoyl-sn-glycerol-3-phosphoethanolamine (DPPE; 1.06 g, 1.5 mmol, 0.9 eq) and triethylamine (600 μL, 0.71 mmol, 2.5 eq) in chloroform. The resulting clear solution was stirred at 60°C for 1 hour (the solution remained clear, and after 1 hour, 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 with Na2SO4, filtered, and the solvent was removed under vacuum. Column chromatography (fast SiO2) with an elution gradient of 2% to 30% methanol in chloroform gave the title compound partially contaminated with triethylamine. The impure fraction was dissolved in chloroform and the organic layer was gently washed with 0.1 M HCl. The organic layer was dried with Na2SO4, filtered, and the solvent was removed under vacuum. This effectively removed triethylamine, and the pure fractions were combined to give the pure product (1.22 g, 1.4 mmol, 91%) as a colorless waxy solid.

[0539] 1H-NMR (400MHz, DMSO-d6): δ = 8.06 (t, J = 5.7Hz, 1H), 7.47-7.22 (m, 6H), 5.15 (dq, J = 8. 3,4.6Hz,1H),5.01(q,J=12.6Hz,2H),4.28(dd,J=12.0,3.2Hz,1H),4.11(dd,J=12.1 ,7.0Hz,1H),4.06-3.92(m,3H),3.82(q,J=6.4Hz,2H),3.33-3.18(m,2H),2.27(dt,J =12.8,5.0Hz,4H),1.50(q,J=6.9Hz,4H),1.32-1.16(m,51H),0.85(t,J=6.7Hz,6H). 31 P-NMR (162MHz, DMSO-d6): δ=-1.4.

[0540] Constructing a block Ala-DPPE

[0541] 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 combined in chloroform / ethanol 1:2 (36 mL). The flask was evacuated and backfilled three times with Ar. An H2 balloon was attached, the flask was evacuated and backfilled three times with H2, and the mixture was stirred at room temperature under positive H2 pressure for 3 h. The solution was filtered through diatomaceous earth 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 diatomaceous earth and washed thoroughly with chloroform / ethanol 2:1. The filtrate was evaporated to dryness to give the product as a pale yellow waxy solid (224 mg, 0.29 mmol, 86%) containing trace amounts of Pd.

[0542] 1 H-NMR (400MHz, DMSO-d6): δ=8.60(br,1H),8.08(br,2H),5.15(br,1H),4.28(d,J=13.2Hz,1H),4.12(dd,J=6.9Hz,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=6.6Hz,6H). 31 P-NMR (DMSO-d6): δ = -1.4.

[0543] MTP(Bn)-a-DPPE(14)

[0544] MDP(Bn) (20.0 mg, 34 μmol) and Ala-DPPE (26.2 mg, 34 μmol, 1.0 eq) were combined in DMAc (0.3 mL) and N,N-diisopropylethylamine (24 μL, 0.14 mmol, 4 eq) and PyBOP (22 mg, 41 μmol, 1.2 eq) were added sequentially. The resulting suspension was stirred at 50 °C for 1 h, after which the mixture became almost clear. Volatiles were removed under vacuum (oil pump, 45 °C) and the mixture was washed once with chloroform. Automated column chromatography (reversed-phase C18; product: C18-silica 1:200; detection: λ = 200–220 nm) was performed using a column chromatography (fast SiO2) with an elution gradient of 15%–40% methanol in chloroform followed by an automated column chromatography (reversed-phase C18; product: C18-silica 1:200; detection: λ = 200–220 nm) using a elution gradient of 30%–80% THF in H2O. After lyophilization, product 14 (8.0 mg, 6 μmol, 18%) was obtained as a white, fluffy solid. HPLC-MS: t[product] = 3.92 min.; m / z = 1327.80 [M+H] + (SIM mode). HPLC-ELSD: t[product] = 3.48 min; 99.2% relative peak area.

[0545] Example 15. MTP-a-chol

[0546]

[0547] Molecular weight: 10¹⁸ Daltons. CLogP = 5.64.

[0548] N-(2-aminoethyl)-cholesterol carbamate constitutes the block.

[0549]

[0550] A solution of cholesterol chloroformate (0.95 g, 2.1 mmol) in 20 mL of DCM was slowly added over approximately 2 hours to a solution of ethylenediamine (2 mL, 14 eq.) in 30 mL of DCM. The reaction was allowed to proceed for another 30 minutes, after which the reaction mixture was evaporated to dryness. The resulting white substance 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.

[0551] 1H NMR (400MHz, CDCl3) δ5.46-5.27(m,1H),4.99(br.s,1H),4.50(br.m,1H),3.22(q,J=5.6Hz,2H), 2.82(t,J=5.9Hz,2H),2.43-2.19(m,2H),2.06-1.75(m,5H),1.64-0.80(m,35H),0.68(s,3H)ppm. 13 C NMR (101MHz, 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. MALDI: m / z = 472.40 (calculated value), measured value: 495.39 (M+Na+). The main peak was observed at m / z = 369.37, which is attributed to the 3,4-elimination product formed in MALDI (not observed in NMR).

[0552] The constituent block can be coupled with N-Boc-L-alanine (CAS[15761-38-3]) via amidation; the Boc group can then be deprotected; finally, the resulting amine-functionalized molecule can be coupled with MDP to obtain MTP-a-chol.

[0553] Example 16. MDP(Bn)-chol

[0554]

[0555] Molecular weight: 1037 Daltons. CLogP = 7.69.

[0556] The N-(2-aminoethyl)-cholesterol aminomethyl ester block (see Example 15) can be coupled with MDP(Bn) via amidation to obtain the molecule MDP(Bn)-chol.

[0557] Examples 17 (MTP-a-DSPE), 18 (MTP(Bn)-a-DSPE), and 19 (MDP(Bn)-DSPE)

[0558]

[0559] MTP-a-DSPE: MW is 1294 Daltons. CLogP = 12.70 and 6.92 (uncharged and charged).

[0560] MTP(Bn)-a-DSPE: MW is 1384 Daltons. CLogP = 14.99 and 9.21 (uncharged and charged).

[0561] MDP(Bn)-DSPE: MW is 1313 Daltons. CLogP = 15.25 and 9.46 (uncharged and charged).

[0562] 1,2-Distearate-sn-glycerol-3-phosphoethanolamine (DSPE; CAS [1069-79-0]) can be linked to N-Boc-L-alanine (CAS [15761-38-3]) via amidation; the Boc group can then be deprotected; finally, the resulting amine-functionalized molecule can be coupled with MDP to obtain MTP-a-DSPE or coupled with MDP(Bn) to obtain MTP(Bn)-a-DSPE.

[0563] Alternatively, DSPE can be coupled with MDP(Bn) via amidation to obtain the molecular MDP(Bn)-DSPE.

[0564] Example 20: Lipophilicity Study

[0565] The CLogP values ​​of the exemplary compounds of this disclosure were evaluated using Perkin Elmer ChemDraw Professional, version 18.0.0231 (4029) software. The results showed values ​​ranging from approximately 4.15 to 18.28. At a physiological pH of approximately 7.4 (i.e., where the COOH and PO3H groups become charged), the CLogP values ​​of two molecules of the invention were between 4 and 5, the CLogP values ​​of three molecules were between 10 and 20, and the CLogP values ​​of the remaining molecules were between 5 and 10.

[0566] Experimentally, the lipophilicity of molecules can also be compared by performing HPLC using the same elution gradient. Molecules with higher affinity to the hydrophobic C18 column material are more lipophilic, resulting in longer retention times. The table below shows that the sample molecules of the present invention (items 1-4) have longer retention times and are therefore more lipophilic compared to the comparative example molecules (items 5 and 6).

[0567] Methods: HPLC-MS (SIM) and HPLC-ELSD were performed on a Phenomenex Kinetex 5 μm EVO C18 100A LC column (50 × 2.1 mm) using the same gradient from eluent A to eluent B, where A = 20 mM NH4HCO2 in H2O with 0.1 v / v% formic acid, and B = 2-propanol / MeCN / H2O 85:15:5, also with 20 mM NH4HCO2 and 0.1 v / v% formic acid.

[0568] Table 3: HPLC-MS or HPLC-ELSD retention times of molecules with C16, C18 and / or benzyl lipophilic units.

[0569]

[0570]

[0571] nd = Not determined; Compound 1 = Mivarutin; Compound 2 = MDP-C18[mur]

[0572] Example 21. Water solubility study

[0573] The solubility of the disclosed compounds in PBS buffer and in water was tested using low concentrations.

[0574] First, the compound was weighed into a vial and PBS buffer (137 mM, 2.7 mM, 10 mM, and 1.8 mM of NaCl, KCl, Na₂HPO₄, and KH₂PO₄, respectively; pH ​​= 7.4) was added to achieve a concentration of 0.2 mg / mL upon complete dissolution. The sample was shaken, allowed to stand for 1 hour, shaken again, and then the appearance of the solution was examined at room temperature (RT). Next, the sample was heated in a 37°C water bath for 1 minute, and the appearance of the solution was examined again. The results are summarized in the table below.

[0575] The tested compounds did not spontaneously dissolve in PBS at room temperature or 37°C. In contrast, the tested comparative compounds spontaneously dissolved under these conditions. Further treatment of the sample solutions with a hot air gun did not dissolve items 4, 5, and 6, while items 2 and 3 yielded turbid solutions after cooling to room temperature.

[0576] Table 4: Solubility test of PBS solution at room temperature and 37°C.

[0577]

[0578] Compound 1 = mifamulin; Compound 2 = MDP-C18[mur]

[0579] Next, the compound was weighed and dissolved in chloroform / methanol. The solution was allowed to stand in a vial to dry, forming a film of the substance. The vial was placed under vacuum to remove trace amounts of organic solvent. Softened water was added so that the concentration of the compound would be 0.3 mM (0.3 mM corresponds to 0.3 mg / mL for a compound with MW = 1000 Daltons) if complete dissolution occurred. The vial was briefly sonicated in a water bath, allowed to stand overnight, and sonicated again (at room temperature). The appearance of the solution at room temperature was examined to assess solubility. The results are summarized in the table below.

[0580] None of the compounds of this disclosure tested spontaneously dissolved in water at room temperature. In contrast, the comparative example compounds tested spontaneously dissolved under these conditions.

[0581] Table 5: Solubility test of aqueous solution at room temperature.

[0582]

[0583] Compound 1 = mifamulin; Compound 2 = MDP-C18[mur]

[0584] Finally, the solubility of Comparative Examples 1 and 2 in PBS (0.01 M, pH = 7.4) and softened water was tested at a level of 1 mg / mL. The same results as shown in the two tables above were obtained at this concentration.

[0585] In summary, these results demonstrate that a series of compounds of the present invention do not spontaneously dissolve in PBS or water at concentrations as low as 0.2 mg / mL (and higher). In contrast, the comparative examples are soluble in PBS or water at concentrations up to at least 0.2 mg / mL or even 1 mg / mL to obtain clear and transparent solutions.

[0586] Due to the low solubility of the disclosed compounds in aqueous solutions, their physicochemical properties are particularly useful in the production of stable HDL-derived NPs. Without being bound by theory, the disclosed compounds are thought to provide improved anchoring of NPs, reduced leakage, and products with greater stability and shelf life.

[0587] Example 22. Degradation using enhanced oxidation test

[0588] Reference compounds MDP and MDP(Bn), as well as 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 to 80°C for 4 hours to achieve rapid molecular degradation by oxidation, simulating slower in vivo oxidation events.

[0589] For the MDP and MDP(Bn) test solutions, the resulting reaction mixture was diluted with acetonitrile and water (1:1), or for the MDP(Bn)-DSPE and MTP(Bn)-a-DPPE solutions, the resulting reaction mixture was diluted with iPrOH, acetonitrile, and water (40:7.5:52.5) ​​(containing 0.1% formic acid and 20 mM ammonium formate). The four diluted samples were analyzed by HPLC-MS. For reference, the four starting materials, as well as MDP-DSPE and MTP-a-DPPE (i.e., the debenzylated reference compounds of the Bn-substituted test molecules), were also analyzed by HPLC-MS.

[0590] For all four test solutions, the unaffected starting compounds were traced. Furthermore, multiple derivatives with masses of +14, +16, +28, +30, and +32 were found, indicating oxidation from CH₂ to the CO moiety (+14) and from CH₂ to the C-OH moiety (+16), as well as combinations of these oxidation events. The tested MDP(Bn)-DSPE and MTP(Bn)-a-DPPE compounds were degraded primarily via oxidation of the Bn group to the benzoate group (+14), followed by hydrolysis of the benzoate (-104). This was confirmed by the predominant presence of debenzylated MDP-DSPE and MTP-a-DPPE compounds as degradation products: confirmed retention times were observed in HPLC, and confirmed masses (relative to the starting compound -90) were observed in MS.

[0591] The results show that the Bn group in the compounds of the present invention has the highest tendency for in vivo oxidative degradation. After oxidation and cleavage of Bn, conventional MDP or MTP groups are formed, and these groups degrade in vivo in a manner similar to other MDPs / MTPs known in the art.

[0592] Example 23. Nanobiosynthesis

[0593] Method 1 - Membrane

[0594] The phospholipids, (pro)drug, and optionally triglycerides or polymers are dissolved (usually in chloroform, ethanol, or acetonitrile). The solution is then evaporated under vacuum to form a membrane of the components. Subsequently, a buffer solution is added to hydrate the membrane and produce a vesicle suspension. Alternatively, the phospholipids, (pro)drug, and optionally triglycerides or polymers are dissolved (usually in chloroform, ethanol, or acetonitrile). The solution is infused or added dropwise to a gently heated buffer solution with stirring until the organic solvent has completely evaporated, producing a vesicle suspension.

[0595] Add apolipoprotein AI (apoA-I) (note that apoA-I may already be in B) to the vesicle suspension generated using A or B – use dropwise addition to avoid denaturation, and sonicate the resulting mixture for 30 minutes using a tip sonicator while thoroughly cooling it in an external ice-water bath. Transfer the resulting solution containing the nanobiomacide and other byproducts to a Sartorius Vivaspin tube with a molecular weight cutoff depending on the expected size of the nanobiomacide (typically a Vivaspin tube with a cutoff of 10,000–100,000 kDa). Centrifuge the tube until ~90% of the solvent volume passes through the filter. Then, add approximately the same volume of buffer as the remaining solution and rapidly spin the tube again until approximately half the volume passes through the filter. Repeat this step twice, then pass the remaining solution through a 0.22 pm polyethersulfone syringe filter to obtain the final nanobiomacide solution.

[0596] Method 2 - Microfluidics

[0597] In an alternative method, the phospholipids, (pro)drug, and optionally triglycerides, cholesterol, sterol 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 microfluidic pump, the contents of the two syringes are mixed using a microvortex platform. The resulting solution, containing the nanobioreactor and other byproducts, is transferred to a Sartorius Vivaspin tube with a molecular weight cutoff depending on the expected particle size (typically a Vivaspin tube with a cutoff of 10,000–100,000 kDa). The tube is centrifuged until ~90% of the solvent volume passes through a filter. Subsequently, approximately equal to the remaining solution volume, a volume of phosphate-buffered saline is added, and the tube is rapidly spun again until approximately half the volume passes through the filter. This step is repeated twice, after which the remaining solution is passed through a polyethersulfone 0.22 pm syringe filter to obtain the final nanobioreactor solution.

[0598] Method 3 - Microfluidic

[0599] In another method according to the invention, microfluidic technology is used to prepare nanoscale assemblies and ultimately nanobiocomposites. A microfluidic is a device for preparing small-particle materials, operating based on the submerged jet principle. When operating the microfluidic to obtain nanoparticles, a high-pressure pump forces a premixed material stream 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 forces, turbulent forces, and cavitation forces are generated within the interaction chamber during microfluidization. The two streams are then recombine at high speed to generate shear. The resulting product can be recycled back to the microfluidic to obtain increasingly smaller particles. The advantages of microfluidization compared to conventional grinding processes include significantly reduced contamination of the final product and ease of scaling up production.

[0600] Formulation 1

[0601] The table below provides details regarding the preparation of HDL-derived nanoparticle formulations. First, DMPC, cholesterol, and the compounds of this invention are dissolved in ethanol (entries A, B, and D) or in an ethanol / DMSO 4 / 1 solution (entries C, E, and F) at given molar ratios, while the protein apoA-1 is additionally dissolved in PBS buffer (pH 7.5). In these formulations, the amount of apoA-1 applied is related to the amount of DMPC (by weight). The organic solution is mixed with the PBS buffer by bringing the organic solution together using a T-joint.

[0602] Purification of the resulting solution was performed using TFF (tangential flow fractionation), thereby removing the organic solvent and dissolving the nanoparticles in PBS. Concentration of the NP solution was achieved by rapid rotational centrifugation. Finally, the HDL-derived nanoparticle solution was filtered through a 0.2-micron Acrodisk PES filter.

[0603] The final HDL-derived nanoparticle solutions typically showed recoveries exceeding 80% for the compounds used (Examples 2, 3, 7, 8, and 13), DMPC, and cholesterol. Recovery was determined by HPLC (for the compounds) and using assays known in the art (for DMPC and cholesterol). The final HDL-derived nanoparticle solutions were at concentrations of approximately 2 to 4 mg / mL based on the compounds.

[0604] Table 6: Formulation compositions of HDL-derived nanoparticles

[0605]

[0606]

[0607] *APO-A1 is used at approximately half the amount of DMPC (by weight (mg)).

[0608] The stability of nanobiopolymers was assessed using dynamic light scattering (DLS).

[0609] The formulations of items A through F were characterized by DLS over an 8-week period. The Z-mean (intensity-weighted average hydrodynamic size) diameters of the nanoparticles in the formulations of Examples 2, 3, 7, and 13 were approximately 20 nm, 30 nm, 20 nm, and 45 nm, respectively. The size of these nanoparticles remained constant over time, as did the particle size dispersibility (PDI). Unloaded particles (item A) were also stable over time (at a diameter of approximately 30 nm). The nanoparticles in the formulation of Example 8 showed a diameter increase from approximately 50 nm to approximately 225 nm from the 2-week to the 5-week time point. By the 5-week time point, the size had stabilized. Using other processing conditions, the material of Example 8 was also most likely formulated as stable particles with a size of 10 to 50 nm.

[0610] The Z-mean diameter and PDI value of nanoparticles in formulations A to F were determined by DLS. Figure 2 As shown in the image.

[0611] Precipitation A (for advanced sonication): DSPC ([816-94-4]; 2.7 mg), cholesterol (0.26 mg), and a compound (0.46 mg) were dissolved in a glass vial using chloroform / methanol (9:1). The solvent was removed by an argon stream, and the resulting membrane was dried under vacuum for >1 h. A solution of apoA-IPBS (6 mL) was added to the vial, followed by sonication in a water bath for 5 min, incubation at 37°C for 20 min, and then advanced sonication for 10 min. The resulting dispersion was centrifuged to remove larger aggregates. The supernatant was transferred to a Vivapin 20 ultrafiltration unit (10 kDa cutoff) and rapidly vortexed to a volume of approximately 1 mL. The resulting dispersion was diluted with PBS and rapidly vortexed to 1 mL, and this step was repeated twice. Finally, the volume was diluted to 2 mL with PBS to obtain the desired nanoparticle solution.

[0612] Precipitation B (for advanced sonication): DMPC (2.7 mg), cholesterol (0.30 mg), and the compound (0.57 mg) were dissolved in a glass vial using chloroform / methanol (9:1). The solvent was removed by an argon stream, and the resulting membrane was dried under vacuum for >1 h. A solution of peptide-2F (apoA-I mimic 18-mer; sequence 257 in Table 2) in PBS (6 mL) was added to the vial, followed by sonication in a water bath for 5 min, incubation at 37 °C for 20 min, and then advanced sonication for 5 min. The resulting dispersion was centrifuged to remove larger aggregates. The supernatant was transferred to a Vivapin 20 ultrafiltration unit (10 kDa cutoff) and rapidly vortexed to approximately 1 mL. The resulting dispersion was diluted with PBS and rapidly vortexed to 1 mL, and this step was repeated twice. Finally, the volume was diluted to 2 mL with PBS to obtain the desired nanoparticle solution.

[0613] T-connector formulation C: DMPC, cholesterol, and the compound were dissolved in ethanol, while apoA-1 was dissolved in PBS buffer (pH 7.5). The organic solution was mixed with the buffer solution using a T-connector mixer. Purification of the resulting solution was performed by TFF (tangential flow filtration) to remove the organic solvent. The sample was concentrated by rapid spin-filtration. Typical recoveries of the compound, DMPC, and cholesterol in the final HDL-derived nanoparticle solution exceeded 75%. The final concentration of the HDL-derived nanoparticle solution was approximately 2 to 4 mg / mL based on the compound.

[0614] Table 7. Formulations of HDL-derived nanoparticles

[0615]

[0616] *Use apoA-I or 2F-peptide in mg / mg phosphoric acid choline (PC); **Use apoA-I in mg / mg compound; # Number-average diameter.

[0617] The above embodiments emphasize that DSPC can be used instead of DMPC (e.g., POPC can also be used), peptide mimics can be used instead of apoA-I, and high levels of the compounds disclosed herein can be introduced. Furthermore, advanced sonication can be used as a processing technique instead of, for example, T-connector mixing or microfluidic mixing.

[0618] Human apoA-I was isolated from human HDL concentrate (Bioresource Technology) following the previously described procedure (Zamanian-Daryoush et al., 2013). Briefly, 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 added to chloroform / methanol solution for defatting. The resulting emulsion was filtered, and the apoA-I precipitate was dried overnight. The protein was refolded in 6M 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 protein characterization and purity were determined by gel electrophoresis and size exclusion chromatography.

[0619] Example 24. Cryo-TEM Measurements of HDL-Derived Nanoparticle Formulations

[0620] The molecular compounds of Examples 2 and 7, namely MDP-DSPE [click] and MDP(Bn)-DSPE [click], were formulated with 1,2-dimyristoyl-sn-glycerol-3-phosphocholine (DMPC; (CAS [18194-24-6])) and APO-A1, as well as varying amounts of cholesterol, to prepare HDL-derived nanoparticle formulations. The relative molar amounts of DMPC, the compounds, and the cholesterol component used are shown below. APO-A1 was used in twice the amount (in mg) of the compounds of the present invention. Dynamic scattering data (DLS) of the processed formulations are also given: the recorded diameters and their errors in parentheses are based on number-mean DLS data. The polydispersity of the particle size recorded by DLS is also shown.

[0621] Table 8: Formulations for cryo-TEM measurements

[0622]

[0623]

[0624] *Use APO-A1 at twice the amount of the compound (by weight (mg)).

[0625] Formulation: DMPC, cholesterol, and the compounds were dissolved in ethanol (entries A, C, and E) or in ethanol / DMSO (entries B, D, and F), while apoA-1 was dissolved in PBS buffer (pH 7.5). The organic solution was mixed with the buffer solution using a T-connector mixer. Purification of the resulting solution was performed by TFF (tangential flow filtration), thereby removing the organic solvent. The sample was concentrated by rapid spin-filtration. Typical recoveries of the compounds used (Examples 2 and 7), DMPC, and cholesterol in the final HDL-derived nanoparticle solution exceeded 75%. Recovery was determined by HPLC (for the compounds) and using assays known in the art (for DMPC and cholesterol). The final concentration of the HDL-derived nanoparticle solution was about 2 to 4 mg / mL based on the compounds (Example 2 or 7).

[0626] Figure 1 Cryo-TEM images of HDL-derived nanoparticles for entries A through F are shown.

[0627] A 50 nm scale bar applies to all six images. When using 0% cholesterol, predominantly spherical disc-like particles are observed (A and B; size approximately 5 to 10 nm). Applying 10% cholesterol mostly reveals slightly extended discs (C and D; length approximately 5 to 25 nm and thickness approximately 5 nm). Using 20% ​​cholesterol reveals distinctly extended worm-like particles. These worm-like particles are approximately 20 to 50 nm in length and approximately 5 nm in thickness (image C). In image F, the worms are approximately 50 to 100 nm in length; similarly, the thickness is approximately 5 nm. In F, worm-like particles are also observed to aggregate into larger stacks.

[0628] The results highlight that the particle size of HDL-derived nanoparticles can be controlled by the cholesterol content of the formulation (comparing C to E and D to F), but can also be controlled by the lipophilicity of the compound and / or the substitution of the R2 position in formula (I) of the compound of the present invention (clear: comparing E to F; less clear: in D, the particles appear to be more elongated than in C).

[0629] Methods: Immediately before sample preparation, a copper grid (Electron Microscopy Sciences) loaded with 200-mesh Lysish carbon was subjected to surface plasmon treatment for 40 seconds using a Cressington 208 carbon coater. Next, 3 μL of a sample solution of HDL-derived nanoparticles was transferred onto the grid. Then, a thin film of the sample solution was vitrified on the grid using an automated vitrification robot (FEI VitrobotMark IV) via plugging vitrification in liquid ethane. The treated film was stored until measurement. Cryo-TEM imaging of the prepared film was performed on a CryoTITAN microscope (Thermo Fisher) equipped with a field emission gun (FEG), a post-column Gatan imaging filter (model 2002), and a rear-mounted GIF 2k×2k Gatan CCD camera (model 794).

[0630] Example 25: In vitro NOD2 activation assay

[0631] The stimulation of human NOD2 (hNOD2) by the compounds disclosed in this article was monitored by HEK-Blue. TM The activation of NF-κB in hNOD2 cells (Invitrogen) was investigated. 50,000 HEK-Blue cells were used. TM hNOD2 cells were seeded in HEK-Blue flat-bottomed tissue culture plates. TM In the detection medium, a specific concentration of the test sample (the compound, first diluted with softened water and then with PBS to the desired concentration) was added to the cells in the tissue culture plate. The cells were incubated overnight at 37°C and 5% CO2. The next day, the supernatant was collected in an ELISA plate and the OD was measured at 620 nm using a spectrophotometer.

[0632] The signal in this experiment was based on NOD2 stimulation using a ligand, which subsequently activated NF-κB and AP-1, leading to the production of SEAP. Then HEK-Blue was used. TM SEAP levels were determined using a test medium (Invitrogen). SEAP hydrolyzes the substrate in the medium, producing a purple / blue color, which was then measured using a microplate reader. OD values ​​were plotted based on the sample concentration and mapped onto [the microplate]. Figure 3 The experimental performance was verified using a cell wall acyl dipeptide (CAS No. [53678-77-6]).

[0633] All tested compounds (APIs) of the present invention were able to activate NOD2. The potency of the compounds in Examples 2, 3, and 7 was comparable. The compounds in Examples 8 and 13 were also able to activate NOD2, but to a lesser extent than the compounds in Examples 2, 3, and 7.

[0634] Example 26: In the B16F10 mouse model, (i) HDL-derived nanoparticles and (ii) immune checkpoint inhibitors were used in the group. Synergistic activity

[0635] In vivo studies of the formulation 1.

[0636] The formulations used in these in vivo studies were prepared by mixing via a T-connector followed by tangential flow filtration (TFF). The same processing and purification steps were applied as those highlighted in the formulations used for cryo-TEM measurements (Table 8).

[0637] The compounds used were those of Examples 2, 3, 7, 8, and 13 (i.e., these examples of substances studied in vivo are collectively referred to as APIs in these descriptions). For the five dosage forms, the following relative proportions of the components were used.

[0638]

[0639] *Relative to compounds

[0640] Plan and Results

[0641] A set of example APIs were formulated as HDL-derived nanoparticles (or nanobioreactors, NB) and their antitumor activity in combination with immune checkpoint inhibitors was screened in a B16F10 homologous mouse tumor model. For this purpose, 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). On the day of injection, cells were harvested and cultured at 1 × 10⁻⁶ cells / day. 6 Cell viability was resuspended at 100 μL / mL in PBS supplemented with 0.5% FBS. During counting, cell viability was assessed using trypan blue solution (0.4%) (Gibco). At the start of the experiment, 1 × 10⁶ cells / mL were resuspended in PBS supplemented with 0.5% fetal bovine serum (FBS). 5 One B16F10 tumor cell was subcutaneously injected into the flank of a 7-week-old female C57BL / 6 mouse (The Jackson Laboratory).

[0642] Seven days after tumor inoculation, mice were randomly assigned to groups with similar average group size (n=10). The average tumor size in each group was 3.26 mm. 3After randomization, mice were ear-cropped and weighed. The doses were then calculated and divided equally. The divided doses were stored at 4°C until use.

[0643] The study consisted of a PBS control group, an immune checkpoint inhibitor group (CI), and six treatment groups. Mice treated with immune checkpoint inhibitors received intraperitoneal injections on days 2, 4, and 8, using doses of 200 μg anti-CTLA-4 (clone, 9H10, BioXcell) and / or 200 μg anti-PD-1 (clone, RMP1-14, BioXcell). The treatment groups consisted of NBs prepared from the compounds of Example 2, NBs from Example 3, NBs from Example 7, NBs from Example 8, and NBs from Example 13, in combination with the immune checkpoint inhibitor therapy (CI) as described above. The doses used in the treatment groups were approximately 9 mg of the discussed MDP / kg (or approximately 27 mg / kg of each API contained in the NB) on days 0, 2, and 4.

[0644] Tumor growth curve plotted on Figure 4 In the AE, and in each figure, the same PBS group and CI group are depicted to allow for comparison between figures.

[0645] Animals treated with PBS or with immune checkpoint inhibitors alone did not show tumor growth inhibition. Animals treated with combination therapy all showed tumor growth inhibition, and for NB (as described in Example 2) Figure 4 A), NB of Example 7 Figure 4 C) or NB (of Example 8) Figure 4 The most obvious results were observed in those groups where D was part of a combination therapy.

[0646] Note that the results of the above combination therapy were obtained using an application formulation containing 10 mol% cholesterol relative to the applied 90 mol% DMPC (see table above), therefore the particle size ranges from approximately 5 nm to a maximum of 10 nm (see table above). Figure 1 Cryo-TEM images C and D).

[0647] Example 27: Single-dose activity of nanobiological agents in the B16F10 mouse model

[0648] The formulations used in these in vivo studies were prepared by mixing via a T-joint followed by tangential flow filtration (TFF). The same processing and purification steps were applied as those highlighted in the formulations used for cryo-TEM measurements (Table 8). The relative proportions of the components used to prepare HDL-derived nanoparticles are shown below.

[0649]

[0650] * Relative to the compound; # Number average diameter.

[0651] Plan and Results

[0652] Two APIs (compounds from Examples 2 and 7) were used to generate a set of different HDL-derived nanoparticle formulations (or nanobiologic formulations; nanobiologics; NB) to determine their potency. The resulting nanobiologic formulations were screened for their single-agent antitumor activity in a B16F10 homologous mouse tumor model. For this purpose, 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). On the day of injection, cells were harvested and cultured at 1 × 10⁻⁶ cells / day. 6 Cell viability was resuspended at 1 × 10⁶ live cells / mL in PBS supplemented with 0.5% FBS. Cell viability was assessed using a cell counter and analyzer (Casy) during counting. At the start of the experiment, 1 × 10⁶ live cells / mL were resuspended in PBS supplemented with 0.5% fetal bovine serum (FBS). 5 One B16F10 tumor cell was subcutaneously injected into the flank of a 7-week-old female C57BL / 6J mouse (Charles River).

[0653] Seven days after tumor inoculation, mice were randomly assigned to groups with similar average group sizes. Each group consisted of 8–10 mice. The average tumor size in each group was 6.33 mm. 3 After randomization, mice were tattooed with numbers on their tails. The doses were then calculated and divided equally. The divided doses were stored at 4°C until use.

[0654] The study consisted of a PBS control group and five treatment groups: NB from Example 2 (formulation entries 1, 3, and 4) and NB from Example 7 (formulation entries 2 and 5). The doses used in the treatment groups were approximately 3 mg of the discussed MDP / kg (or approximately 9 mg / kg of each API contained in the NB) on days 0, 2, and 4. Tumor size was measured at set time points throughout the study. Tumor growth curves were plotted on... Figure 5 In the AC, and in each figure, the same PBS group is depicted to allow for comparison between figures. Except for NB (formulation entry 1) of Example 2, all nanobioreactors clearly showed a reduction in tumor growth compared to the PBS control group. For Example 2 ( Figure 5 Example A) and Example 7 ( Figure 5 Of the two, the nanobiological agents with 20 mol% cholesterol performed best. Figure 5C shows that the NB of Example 2 is slightly better than the NB of Example 7 (using the formulations of Entries 4 and 5, respectively).

[0655] Note the attached diagram. Figure 5 In Example A, the formulation of Example 2 containing 10 mol% cholesterol (entry 1 in the table above) showed lower single-dose activity (compared to PBS). Surprisingly, in contrast, the formulations of Example 2 containing 20 mol% cholesterol (entries 3 and 4 in the table above) showed extremely enhanced tumor suppression. This was also observed with the formulation of Example 7. Figure 5 (B). Clearly, the size of the formed particles plays a decisive role in the activity of the prepared HDL-derived nanoparticles: a 10% cholesterol formulation yields spherical or slightly stretched disc-like particles with sizes of 5 to 10 nm, while a 20% cholesterol formulation yields elongated worm-like particles with lengths of approximately 15 to 50 nm and thicknesses of approximately 5 nm (comparative). Figure 1 Figures C and E in the figure; frozen TEM data).

[0656] The disclosed HDL-derived nanoparticles possess specific sizes, and these characteristics are particularly useful in the production of stable and potent HDL-derived NPs. Without being bound by theory, it is believed that the disclosed HDL-derived nanoparticles provide improved (multivalent) presentation of MDP (or MDP(Bn) or MTP or MTP(Bn)) fractions to cells, significantly enhancing their potency.

[0657] Implementation Plan

[0658] 1. A compound of formula (I) or a pharmaceutically acceptable salt thereof:

[0659]

[0660] in:

[0661] R 1 -H or -C(O)-R X ;

[0662] R 2 and R 3 Each is independently selected from the group consisting of -H, alkyl, alkylene-aryl, -C(O)-alkyl, and -C(O)-aryl;

[0663] R 4 R 5 and R 5’ Each is an alkyl group;

[0664] R 6 and R 11 Each is independently -H or alkyl;

[0665] R7 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 chains,

[0666] -(CR 10 R 10 )2-OP(O)(OH)-O-alkylene-C(R 10 (OR) Z )-alkylene-OR Z Or -γ-triazolyl-L;

[0667] R Z C 8-30 Fatty acids or -C(O)-C 16-30 fatty acid chains;

[0668] Y is an alkylene group;

[0669] L is selected from fatty acid chains, -alkylene-C(O)-W, -alkylene-OC(O)-W, -alkylene-N-(alkylene-C(O)-NR) 11 -alkylene-NR 11 The group consisting of -C(O)-W)2 and -alkylene-N-(alkylene-C(O)-W)2;

[0670] W represents a fatty acid chain, -O-alkylene-C(H)(OR) 8 )-alkylene-OR 9 Phospholipids or sterols;

[0671] R 8 and R 9 Each independently as R X or -C(O)-R X ;

[0672] R 10 R 22 R 33 R 33’ R 44 R 44’ R 55 and R 55’ Each independently is H or R A ;

[0673] R X It consists of fatty acid chains;

[0674] Each of the aforementioned alkyl, alkylene, alkylene-aryl, aryl, and triazolyl groups is optionally surrounded by one or more R groups. A Replace, where R A Each time it appears, it is independently selected from the following groups: halogen, 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 (where X is 0, 1, or 2), aryl, arylalkyl, heteroaryl, heteroarylalkyl, cycloalkyl, heterocycloalkyl, and R B ;

[0675] R C and R D Each occurrence is independently selected from hydrogen, alkyl, haloalkyl, -C(O)R B and -C(O)OR B The group formed; or R C and R D Together with the nitrogen they are attached to, they form optional R A Substituted heterocycles; and

[0676] R B The alkyl, alkenyl, or alkynyl groups may be optionally substituted with one or more fluorine groups;

[0677] Where R 7 C 9-30 When fatty acid chains are in the form of R 2 It is -H.

[0678] 2. The compound of embodiment 1, wherein the compound of formula (I) is the compound of formula (IA) or a pharmaceutically acceptable salt thereof:

[0679]

[0680]

[0681] 3. The compound of embodiment 1 or 2, wherein R 2It is -H or benzyl.

[0682] 4. A compound according to any one of embodiments 1-3, wherein R 2 It is -H.

[0683] 5. A compound according to any one of embodiments 1-4, wherein R 10 R 22 R 33 R 33’ R 44 R 44’ R 55 and R 55’ Each is -H.

[0684] 6. A compound according to any one of embodiments 1-5, wherein R 4 It is an alkyl group.

[0685] 7. A compound according to any one of embodiments 1-6, wherein R 4 It is a methyl group.

[0686] 8. A compound of any one of embodiments 1-7, wherein R 3 and R 6 All are -H.

[0687] 9. A compound of any one of embodiments 1-8, wherein Y is optionally -C(O)N(R) C (R) D Alkylene substituted with alkylene.

[0688] 10. The compound of embodiment 9, wherein Y is -CH2- or

[0689] 11. The compound of embodiment 10, wherein Y is -CH2-.

[0690] 12. A compound of any one of embodiments 1-11, wherein R 7 It is -γ-triazolyl-L;

[0691] 13. A compound of any one of embodiments 1-12, wherein the compound of formula (I) is a compound of formula (II) or a pharmaceutically acceptable salt thereof:

[0692]

[0693] in:

[0694] X1 is -N- and X2 is -C-; or X1 is -C- and X2 is -N-.

[0695] 14. The compound of embodiment 13, wherein the compound of formula (I) is a compound of formula (IIA) or a pharmaceutically acceptable salt thereof:

[0696]

[0697] 15. The compound of embodiment 13 or 14, wherein Y is C 1-6 Alkylene.

[0698] 16. A compound of any one of embodiments 13-15, wherein R A It is -H.

[0699] 17. A compound of any one of embodiments 1-16, wherein L is selected from 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 group consists of -C(O)-W)2 and -CH2CH2-N-(CH2CH2-C(O)-W)2.

[0700] 18. The compound of embodiment 17, wherein L is C 12-18 Fatty acid chains.

[0701] 19. The compound of embodiment 17, wherein L is -CH2(CH2CH2)8-CH3.

[0702] 20. A compound of any one of embodiments 1-19, wherein W is C 8-30 Fatty acid chains.

[0703] 21. A compound of any one of embodiments 1-19, wherein W is C 12-18 Fatty acid chains.

[0704] 22. A compound of any one of embodiments 1-19, wherein W is:

[0705]

[0706] 23. The compound of embodiment 22, wherein R X and R X’ Each independently is -C 8-30 Fatty acid chains.

[0707] 24. The compound of embodiment 22 or 23, wherein R X and R X’ Each independently is C 12-18 Fatty acid chains.

[0708] 25. The compound of embodiment 24, wherein R X and R X’ All are -(CH2CH2)8-CH3.

[0709] 26. A compound of any one of schemes 1-19, wherein W is cholesterol:

[0710]

[0711] 27. A 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.

[0712] 28. The compound of embodiment 27, wherein W is a phospholipid having the following structure or a pharmaceutically acceptable salt thereof:

[0713]

[0714] Where R X and R X’ Each independently is C 8-30 Fatty acid chains.

[0715] 29. The compound of embodiment 28, wherein R X and R X’ Each independently is C 12-18 fatty acid.

[0716] 30. The compound of implementation scheme 28 or 29, wherein the fatty acid is saturated.

[0717] 31. The compound of embodiment 28, wherein R X and R X’ All are -(CH2CH2)8-CH3.

[0718] 32. A compound of any one of embodiments 1-11, wherein R 7 -C(H)(C(O)NH2)-C5 alkylene-N(R) 11 )-C(O)-C 17-30 fatty acid.

[0719] 33. A compound of any one of embodiments 1-11, wherein R 7 for

[0720] -CH2CH2-OP(O)(OH)-O-CH2-C(H)(OR Z )-CH2-OR Z .

[0721] 34. A compound of any one of embodiments 1-33, wherein the compound is selected from the group consisting of:

[0722]

[0723]

[0724]

[0725] 35. A nanobiocomposition comprising nanoparticles derived from high-density lipoprotein (HDL), wherein the nanoparticles comprise a compound of formula (I) or a pharmaceutically acceptable salt thereof:

[0726]

[0727] in:

[0728] R 1 -H or -C(O)-R X ;

[0729] R 2 and R 3 Each is independently selected from the group consisting of -H, alkyl, alkylene-aryl, -C(O)-alkyl, and -C(O)-aryl;

[0730] R 4 R 5 and R 5’ Each is an alkyl group;

[0731] R 6 and R 11 Each is independently -H or alkyl;

[0732] R 7 For fatty acid chains, -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 -γ-triazolyl-L;

[0733] Y is an alkylene group;

[0734] L is selected from fatty acid chains, -alkylene-C(O)-W, -alkylene-OC(O)-W, -alkylene-N-(alkylene-C(O)-NR) 11 -alkylene-NR11 The group consisting of -C(O)-W)2 and -alkylene-N-(alkylene-C(O)-W)2;

[0735] W represents a fatty acid chain, -O-alkylene-C(H)(OR) 8 )-alkylene-OR 9 Phospholipids or sterols;

[0736] R 8 and R 9 Each independently as R X or -C(O)-R X ;

[0737] R 10 R 22 R 33 R 33’ R 44 R 44’ R 55 and R 55’ Each independently is H or R A ;

[0738] R X It consists of fatty acid chains;

[0739] Each of the aforementioned alkyl, alkylene, alkylene-aryl, aryl, and triazolyl groups is optionally surrounded by one or more R groups. A Replace, where R A Each time it appears, it is independently selected from the following groups: halogen, 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 (where X is 0, 1, or 2), aryl, heteroaryl, heteroarylalkyl, cycloalkyl, heterocycloalkyl, and R B ;

[0740] R C and R D Each occurrence is independently selected from hydrogen, alkyl, haloalkyl, -C(O)RB and -C(O)OR B The group formed; or R C and R D Together with the nitrogen they are attached to, they form optional R A Substituted heterocycles; and

[0741] R B It can be an alkyl, alkenyl, or alkynyl group optionally substituted with one or more fluorine groups.

[0742] 36. A nanobiocomposition comprising nanoparticles derived from high-density lipoprotein (HDL), wherein the nanoparticles comprise a compound of any one of embodiments 1-34.

[0743] 37. The nanobiocomposition of embodiment 35 or 36, wherein the HDL-derived nanoparticles comprise one or more phospholipids.

[0744] 38. The nanobiocomposition of embodiment 37, wherein the 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.

[0745] 39. The nanobiocomposition of embodiment 35 or 36, comprising phospholipids selected from the group consisting of 1,2-dimyristoyl-sn-glycerol-3-phosphatidylcholine (DMPC) and 1-palmitoyl-2-oleoyl-sn-glycerol-3-phosphocholine (POPC), and hemolysin selected from the group consisting of 1-myristoyl-2-hydroxy-sn-glycerol-phosphocholine (MHPC) and 1-palmitoyl-2-hydroxy-sn-glycerol-3-phosphocholine (PHPC).

[0746] 40. The nanobiocomposition of any one of embodiments 35-39, wherein the HDL-derived nanoparticles contain apoA-I or a peptide mimic of apoA-I.

[0747] 41. The nanobiocomposition of any one of embodiments 35-40, wherein the HDL-derived nanoparticles further comprise one or more triglycerides, fatty acid esters, hydrophobic polymers, sterol esters, or combinations thereof.

[0748] 42. The nanobiocomposition of any one of embodiments 35-41, wherein the HDL-derived nanoparticles further comprise cholesterol.

[0749] 43. The nanobiocomposition 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.

[0750] 44. The nanobiocomposition of embodiment 43, wherein the HDL-derived nanoparticles contain one or more phospholipids and cholesterol in a molar ratio of about 1:0.2.

[0751] 45. The nanobiocomposition of any one of embodiments 1-44, wherein the nanoparticles derived from HDL are nanodiscs or nanospheres.

[0752] 46. ​​The nanobiocomposition of embodiment 45, wherein the diameter of the nanodisc or nanosphere is from about 8 nm to about 400 nm.

[0753] 47. A pharmaceutical composition comprising a compound of any one of embodiments 1-34 or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.

[0754] 48. A method for treating cell proliferation disorders in patients in need, comprising administering to the patient a therapeutically effective amount of the nanobiocomposition of any one of embodiments 35-46.

[0755] 49. The method of implementation plan 48, wherein the cell proliferation disease is cancer.

[0756] 50. The method of implementation scheme 49, wherein the cancer is selected from the group consisting of: bladder cancer, vascular cancer, bone cancer, brain cancer, breast cancer, cervical cancer, chest cancer, colon cancer, endometrial cancer, esophageal cancer, eye cancer, head cancer, kidney cancer, liver cancer, lymph node cancer, lung cancer, oral cancer, neck cancer, ovarian cancer, pancreatic cancer, prostate cancer, rectal cancer, colorectal cancer, skin cancer, stomach cancer, testicular cancer, laryngeal cancer, thyroid cancer, urothelial carcinoma, and uterine cancer.

[0757] 51. The method of implementation scheme 49, wherein the cancer is selected from the group consisting of breast cancer, prostate cancer, melanoma, colorectal cancer, lung cancer, pancreatic cancer and glioblastoma.

[0758] 52. The method of any one of embodiments 48-51, wherein the method further comprises co-administering a cancer drug with a nanobiocomposition as a combination therapy.

[0759] 53. A method for treating sepsis in patients in need, comprising administering to the patient a therapeutically effective amount of any one of embodiments 35-46 of the nanobiocomposition.

[0760] 54. The method of implementation plan 53, wherein the patient suffers from sepsis associated with a bacterial, viral, or fungal infection of the lungs, abdomen, kidneys, or bloodstream.

[0761] 55. The method of any one of embodiments 48-54, wherein the nanobiocomposition promotes a hyperresponsive innate immune response in patients in need.

[0762] 56. The method of implementing 55, wherein a hyperresponsive innate immune response is promoted for at least about 7 to about 30 days.

[0763] 57. The method of implementation plan 55, wherein a hyperresponsive innate immune response is promoted for at least 30 to 100 days.

[0764] 58. The method of implementing 55, wherein a hyperresponsive innate immune response is promoted for more than 100 days and up to 3 years.

[0765] 59. The method of embodiment 55, wherein the nanobiocomposite is administered once and wherein a hyperresponsive innate immune response is promoted for at least 30 days.

[0766] 60. The method of embodiment 55, wherein the nanobiocomposite is administered at least once a day on each day of a multiple dosing regimen, and wherein a hyperresponsive innate immune response is promoted for at least 30 days.

[0767] 61. The method of any one of embodiments 48-58, wherein the nanobiocomposite is administered to a patient in a treatment regimen comprising two or more administrations to generate accumulation of the drug in myeloid cells, bone marrow progenitor cells and hematopoietic stem cells in the bone marrow, blood and / or spleen.

[0768] 62. The method of any one of embodiments 48-61, wherein the nanobiocomposition is administered intravenously or intra-arterially.

[0769] 63. A method for activating a NOD2 receptor in a subject in need, comprising administering to the subject an effective amount of the nanobiocomposition of any one of embodiments 35-46.

[0770] 64. A process for manufacturing a nanobiocomposition according to any one of embodiments 35-46, said process comprising:

[0771] a) Forming a lipid membrane comprising, under conditions conducive to effective lipid membrane formation: i) a compound of 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

[0772] b) Dissolve the lipid membrane in a solvent to form a lipid solution; and contact the lipid solution with apoA-I or a peptide mimic of apoA-I under conditions that effectively form HDL-derived nanoparticles containing any of the compounds in embodiments 1-34.

[0773] 65. A nanobiocomposition prepared according to embodiment 64.

[0774] 66. A kit comprising a nanobiocomposition of any one of embodiments 35-46.

[0775] 67. The method of claim 52, wherein the cancer drug is a checkpoint inhibitor.

[0776] 68. The method of claim 67, wherein the checkpoint inhibitor is selected from anti-PD-1 antibody, anti-CTLA-4 antibody, and combinations thereof.

Claims

1. A compound of Formula (I): or a pharmaceutically acceptable salt thereof, wherein:

2. The compound of claim 1, wherein the compound of Formula (I) is a compound of Formula (IA): or a pharmaceutically acceptable salt thereof. R 1 is -H; R 2 is -H or benzyl; R 3 is -H; R 4 , R 5 , and R 5’ are each -C 1-6 alkyl; R 6 and R 11 each independently -H; R 7 -Y-triazolyl-L; Y is -C 1-6 alkylene; L is selected from -C 15-20 alkyl, -C 1-6 alkylene-C(O)-W, -C 1-6 alkylene-O-C(O)-W, -C 1-6 alkylene-N-(C 1-6 alkylene-C(O)-NR 11 -C 1-6 alkylene-NR 11 -C(O)-W)2or -C 1-6 alkylene-N-(C 1-6 alkylene-C(O)-W)2; W is -C 12-18 alkyl, -O-C 1-6 alkylene-C(H)(OR 8 -C 1-6 alkylene-OR 9 , cholesterol or a phospholipid having the structure; wherein the cholesterol is Y Q1 , Y Q2 , and Y Q3 are each independently -C 1-6 alkylene; R 8 and R 9 each independently is R X or -C(O)-R X ; R 22 , R 33 , R 33’ , R 44 , R 44’ , R 55 , and R 55’ are each independently H; R X and R X’ each independently is -C 15-20 alkyl; wherein each of the foregoing -C 1-6 alkyl and -C 1-6 alkylene is optionally substituted with one or more R A , wherein R A is independently, at each occurrence, selected from hydrogen, halogen, cyano, hydroxyl, or -C(O)N(R C )(R D ); R C and R D is hydrogen.

5. The compound of claim 1, wherein Y is -CH2-.

6. The compound of claim 1, wherein the compound of Formula (I) is a compound of Formula (II): or a pharmaceutically acceptable salt thereof, 3. The compound of claim 1, wherein R 2 is -H.

4. The compound of claim 1, wherein R 4 is methyl. wherein: X1is -N- and X2is -C-; or X1is -C- and X2is -N-.

7. The compound of claim 6, wherein the compound of Formula (II) is a compound of Formula (IIA): or a pharmaceutically acceptable salt thereof.

9. The compound of claim 1, wherein L is -CH2-C(O)-W.

10. The compound of claim 1, wherein W is:

12. The compound of claim 1, wherein W is a phospholipid having the structure: wherein 8. The compound of claim 6, wherein R A is -H.

13. The compound of claim 12, wherein W is a phospholipid having the structure: or a pharmaceutically acceptable salt thereof; wherein R X and R X’ each independently is -C 15-20 alkyl.

11. The compound of claim 1, wherein R X and R X’ are each -(CH2CH2)8-CH3.

15. The compound of claim 1, selected from the group consisting of: Bn is benzyl. Y Q1 , Y Q2 , and Y Q3 are each independently -C 1-6 alkylene and R X and R X’ are each independently -C 15-20 alkyl.

16. The compound of claim 6, wherein the compound of Formula (II) is a compound of Formula (II-1): or a pharmaceutically acceptable salt thereof.

17. The compound of claim 15, wherein the compound is: wherein R X and R X’ each independently is -C 15-20 alkyl.

14. The compound of claim 13, wherein R X and R X’ are each -(CH2CH2)8-CH3. or a pharmaceutically acceptable salt thereof. and pharmaceutically acceptable salts thereof, wherein, 18. The compound of claim 15, wherein the compound is: or a pharmaceutically acceptable salt thereof.

19. The compound of claim 15, wherein the compound is: or a pharmaceutically acceptable salt thereof.

20. The compound of claim 15, wherein the compound is: or a pharmaceutically acceptable salt thereof.

21. A nanobiological composition comprising a nanoparticle derived from high density lipoprotein (HDL), wherein the nanoparticle comprises one or more phospholipids, and a compound of any one of claims 1-20.

22. The nanobiological composition of claim 21, 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.

23. The nanobiological composition of claim 21, wherein the phospholipid is selected from the group consisting of l,2-dimyristoyl-sn-glycero-3-phosphatidylcholine (DMPC) and l-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC).

24. The nanobiological composition of claim 23, comprising a lysolipid selected from the group consisting of l-myristoyl-2-hydroxy-sn-glycero-phosphocholine (MHPC) and l-palmitoyl-2-hydroxy-sn-glycero-3-phosphocholine (PHPC). ​ ​ ​ ​ ​ 25. The nanobiological composition of claim 21, wherein the HDL-derived nanoparticle further comprises apoA-I or a peptidomimetic of apoA-I.

26. The nanobiological composition of claim 21, wherein the HDL-derived nanoparticle further comprises cholesterol.

27. The nanobiological composition of claim 26, wherein the HDL-derived nanoparticle comprises one or more phospholipids and cholesterol in a molar ratio in the range of 1:0.05 to 1:0.

25.

28. A pharmaceutical composition comprising the compound of any one of claims 1-20, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.

29. Use of the nanobiological composition of any one of claims 21-27 or the pharmaceutical composition of claim 28 in the manufacture of a medicament for treating cancer.

30. The use of claim 29, wherein the cancer is selected from the group consisting of bladder cancer, blood vessel cancer, bone cancer, brain cancer, breast cancer, cervical cancer, chest cancer, colon cancer, endometrial cancer, esophageal cancer, eye cancer, head cancer, kidney cancer, liver cancer, lymph node cancer, lung cancer, mouth cancer, neck cancer, ovarian cancer, pancreatic cancer, prostate cancer, rectal cancer, colorectal cancer, skin cancer, stomach cancer, testicular cancer, throat cancer, thyroid cancer, urothelial cancer, and uterine cancer.

31. Use of the nanobiological composition of any one of claims 21-27 or the pharmaceutical composition of claim 28 in the manufacture of a medicament for treating sepsis.

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