Sphingolipid-loaded nano biological preparation for immunomodulation

Abstract

Provided herein is a sphingolipid loaded nanobiologic, comprising: an apolipoprotein A-1 (apoA-1) or a peptidomimetic of apoA-1; sphingolipid; the present invention relates to a nanobiological composition, and cholesterol, and uses thereof, for example, including methods of stimulating innate immune modulation of a training immune response in a subject, and methods of treating cancer, comprising administering the nanobiological composition.

Description

[0001] This application claims priority to U.S. Provisional Application No. 63 / 326,191, filed on March 31, 2022, the contents of which are incorporated herein by reference in their entirety for all purposes.

[0002] Statement of Government Interest

[0003] This invention was made in part with U.S. government support under Grant Nos. R01 CA220234, R01HL144072, and P01 HL131478 awarded by the National Institutes of Health. The U.S. government has certain rights in this invention.

[0004] Description of the electronically submitted text file

[0005] This application contains a sequence listing that has been submitted electronically as an XML file named 27527-0207WO1_SL_ST26.xml. The XML file was created on March 30, 2023 and is 490,269 bytes in size. The material in the XML file is incorporated herein by reference in its entirety. Background Art

[0006] The immune system plays an important role in the pathophysiology of major diseases such as atherosclerosis, diabetes and cancer. However, most of the immunotherapy strategies currently under development focus on any one of effector molecules such as cytokines or T lymphocytes, which are cells from the adaptive immune system. In autoimmune and autoinflammatory diseases, anti-cytokine therapy can successfully neutralize bioactive cytokines, and the most strongly used immunotherapy in cancer patients includes the application of checkpoint inhibitor drugs. Although the innate immune system has long been considered to lack memory, recent studies have shown that innate immune cells undergo metabolic and epigenetic rewiring (epigenetic rewiring) in a process called "training immunity" to regulate their functional programs.

[0007] Sphingolipids are an extended family of bioactive lipids that are ubiquitous in eukaryotes and possess immunomodulatory properties. These lipid molecules regulate fundamental cellular processes, including growth, adhesion, migration, apoptosis, and aging. In addition, sphingolipids play a key role in shaping the immune response.

[0008] There is a need for therapeutic agents that engage the innate immune system, compositions thereof, and methods of use thereof. Summary of the Invention

[0009] The present disclosure relates to sphingolipid-loaded nanobiologics and methods of using the same, for example, in the treatment of cancer.

[0010] In an embodiment, the present disclosure provides a nanobiologic composition comprising:

[0011] (a) apolipoprotein AI (apoA-I) or a peptide mimetic of apoA-I; and

[0012] (b) Sphingolipids.

[0013] In an embodiment, the present disclosure provides a nanobio composition comprising:

[0014] (a) apolipoprotein AI (apoA-I) or a peptide mimetic of apoA-I;

[0015] (b) sphingolipids; and

[0016] (d) cholesterol; and

[0017] wherein the composition is a nanoparticle having a diameter of about 8 nm to about 150 nm. In an embodiment, the nanobiocomposition further comprises a phospholipid.

[0018] In an embodiment, the present disclosure provides a nanobio composition comprising:

[0019] (a) apolipoprotein AI (apoA-I) or a peptide mimetic of apoA-I;

[0020] (b) sphingolipids;

[0021] (c) phospholipids; and

[0022] (d) cholesterol;

[0023] wherein the sphingolipids are present as about 1-100 mol % of the total lipid composition of the nanobiocomposition; and

[0024] The composition is a nanoparticle having a diameter of about 8 nm to about 150 nm.

[0025] In an embodiment, the sphingolipid is selected from the group consisting of ceramide, sphingomyelin, dihydroceramide, glucosylceramide, sphingosine, sphingosine-1-phosphate, galactosylceramide, ceramide-1-phosphate, lactosylceramide, and mixtures thereof.

[0026] In an embodiment, the sphingolipid is of formula (I):

[0027]

[0028] or a pharmaceutically acceptable salt thereof, wherein:

[0029] R 1is a saturated aliphatic chain or an unsaturated aliphatic chain;

[0030] R 2 Is H or R 4 , or C(O)R 4 ;

[0031] R 3 Is H, sugar, P(O)(OH)2, or P(O)(OH)O(CH2) n R 5 , where n = 1-10;

[0032] R 4 is a saturated aliphatic chain or an unsaturated aliphatic chain; and

[0033] R 5 It is H, NH2, N(CH3)3 + , OH, or sugar.

[0034] In an embodiment, the sphingolipid is of formula (IA):

[0035]

[0036] where R 1 and R 2 Defined in this article.

[0037] In an embodiment, the sphingolipid is of formula (II):

[0038]

[0039] where R 1 and R 2 Defined in this article.

[0040] In an embodiment, the sphingolipid is of formula (II-A):

[0041]

[0042] where R 1 and R 2 Defined in this article.

[0043] In an embodiment, the sphingolipid of formula (II) is a sphingolipid of formula (II-B):

[0044]

[0045] where R 1 and R 2 Defined in this article.

[0046] In an embodiment, the nanobiocomposition comprises human apolipoprotein AI (apoA-I).

[0047] In embodiments, the nanobiocomposition is disc-like in shape.

[0048] In embodiments, the nanobiocomposition is spherical in shape.

[0049] In embodiments, the nanobiocomposition is suitable for intravenous or intraarterial administration.

[0050] In embodiments, the present disclosure provides methods for treating cancer in a subject in need thereof, comprising administering to the subject an effective amount of a nanobio composition provided herein.

[0051] In an embodiment, the present disclosure provides methods for treating atherosclerosis in a subject in need thereof, comprising administering to the subject an effective amount of a nanobio composition provided herein.

[0052] In embodiments, the present disclosure provides methods for preventing organ or tissue rejection in a subject in need thereof, comprising administering to the subject an effective amount of a nanobio composition provided herein. BRIEF DESCRIPTION OF THE DRAWINGS

[0053] Figure 1A Schematic diagram of the major sphingolipid classes and their metabolic interconversions. Yellow boxes indicate the sphingolipid classes incorporated into nanobiologics.

[0054] Figure 1B Schematic diagram of an in vitro trained immunoassay using sphingolipid-loaded nanobiologics.

[0055] Figure 1C is a schematic diagram of a sphingolipid-loaded nanobio composition of the present disclosure.

[0056] Figure 1D and Figure 1E is a graph depicting the results of the assay described in Example 2, wherein the sphingolipid-loaded nanobiological preparation prepared in Example 1 was used alone ( Figure 1D ) or in combination with HKCA ( Figure 1E ) were used to stimulate PBMC for 24 hours. After a five-day rest period, cells were restimulated with LPS for 24 hours and cytokine production was measured in the supernatant by ELISA (n=3). Data are presented as ( Figure 1D ) or compared to untrained (RPMI) PBMCs in the log2 fold change ( Figure 1E ) The log2 fold change compared to HKCA-trained PBMCs in . p values ​​were calculated using one-way ANOVA with Dunnett's post hoc test.

[0057] Figure 2Shown are tumor growth curves in the B16F10 mouse melanoma model after treatment with PBS or the nanobiocomposition described in Example 3.

[0058] Figure 3A Shown are (A) lactate dehydrogenase (LDH) measurements of PBMCs treated with sphingolipid-nanobio for 24 hours.

[0059] Figures 3B-3E showed that the sphingolipid-nanobacterial preparation alone ( Figure 3B , Figure 3C ) or in combination with heat-killed Candida albicans (HKCA) ( Figure 3D , Figure 3E Cytokine production was measured in the supernatant after PBMCs were stimulated with LPS for 24 hours (n=3) and restimulated with LPS for 24 hours after a five-day rest period. Data are presented as fold change compared to untrained (RPMI) PBMCs in (B, C) or fold change compared to HKCA-trained PBMCs in (D, E). *p<0.05, **p<0.01, ***p<0.001, p values ​​were calculated using one-way ANOVA with Dunnett's post hoc test.

[0060] Figure 4 Shown are blood chemistry analyses performed in the studies described in Example 2. ALP = alkaline phosphatase, AST = aspartate aminotransferase, ALT = alanine aminotransferase, BUN = blood urea nitrogen, n = 4.

[0061] Figure 5 show 89 Pharmacokinetics of Zr radiolabeled nanobiologic formulation #13.

[0062] Figure 6 show 89 24-hour biodistribution of Zr radiolabeled nanobioformulation #13 by ex vivo gamma counting (n=5).

[0063] Figure 7 Shown are bone marrow analyzed by flow cytometry 24 hours after intravenous administration of nanobioformulation #13 loaded with the lipophilic fluorophore DiOC18 to a B16F10 melanoma mouse model (n=5).

[0064] definition

[0065] For convenience, certain terms employed in the specification, examples, and claims are collected here. Unless otherwise defined, all technical and scientific terms used in this disclosure have the same meaning as commonly understood by one of ordinary skill in the art to which this disclosure belongs.

[0066] The term "about" means a range (e.g., plus or minus 10% of the value) when it is immediately before a numerical value. For example, "about 50" can mean 45 to 55, "about 25,000" can mean 22,500 to 27,500, etc., unless the context of the present disclosure indicates otherwise, or the context of the present disclosure is inconsistent with such an explanation. For example, in a numerical list such as "about 49, about 50, about 55, ...", "about 50" means a range extending to less than half of the interval between the previous value and the subsequent value, for example, more than 49.5 to less than 50.5. In addition, the phrase "less than about" value or "greater than about" value should be understood according to the definition of the term "about" provided herein. Similarly, the term "about" when preceding a series of numerical values ​​or a range of values ​​(e.g., "about 10, 20, 30" or "about 10-30") refers to all values ​​in the series or the endpoints of the range, respectively.

[0067] As used herein, the phrase "pharmaceutically acceptable" refers to those compounds, materials, compositions, and / or dosage forms which are, within the scope of sound medical judgment, suitable for use in contact with the tissues of human beings and animals without excessive toxicity, irritation, allergic response, or other problem or complication, commensurate with a reasonable benefit / risk ratio.

[0068] As used herein, the term "conservative substitution" refers to exchanging one amino acid from the following groups of amino acids for another: (i) aliphatic amino acids (alanine, valine, leucine, and isoleucine); (ii) amino acids having a hydroxyl group (serine and threonine); (iii) acidic amino acids (glutamic acid and aspartic acid); (iv) amino acids having an amide side chain (asparagine and glutamine); (v) basic amino acids (lysine and arginine); and (vii) amino acids having an aromatic side chain (phenylalanine, tyrosine, and tryptophan).

[0069] "Salts" include those obtained by reacting a compound that acts as a base with an inorganic or organic acid to form a salt, or those obtained by reacting a compound that acts as an acid with an inorganic or organic base to form a salt. "Salts" include derivatives of the active agent, wherein the active agent is modified by preparing an acid or base addition salt thereof. 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, ethanesulfonic acid, 2-hydroxyethylsulfonic acid, tartaric acid, ascorbic acid, pamoic acid, bismethylene salicylic acid, ethanedisulfonic acid, gluconic acid, citraconic acid, aspartic acid, stearic acid, palmitic acid, EDTA, glycolic acid, p-aminobenzoic acid, glutamic acid, benzenesulfonic acid, 4-chlorobenzenesulfonic acid, 2-naphthalenesulfonic acid, camphorsulfonic acid, p-toluenesulfonic acid, sulfates, nitrates, phosphates, perchlorates, borates, acetates, benzoates, hydroxynaphthoates, glycerophosphates, and ketoglutarates, among others. Base addition salts include, but are not limited to, ethylenediamine, N-methyl-glucamine, lysine, arginine, ornithine, choline, N,N'-dibenzylethylenediamine, chloroprocaine, diethanolamine, procaine, N-benzylphenethylamine, diethylamine, piperazine, tris-(hydroxymethyl)-aminomethane, tetramethylammonium hydroxide, triethylamine, dibenzylamine, ephenamine, dehydroabietylamine, N-ethylpiperidine, benzylamine, tetramethylammonium, tetraethylammonium, methylamine, dimethylamine, trimethylamine, ethylamine, basic amino acids such as lysine and arginine, and dicyclohexylamine. Examples of metal salts include lithium, sodium, potassium, magnesium, and calcium salts. Examples of ammonium salts and alkylated ammonium salts include ammonium, methylammonium, dimethylammonium, trimethylammonium, ethylammonium, hydroxyethylammonium, diethylammonium, butylammonium, and tetramethylammonium salts. Examples of organic bases include lysine, arginine, guanidine, diethanolamine, and choline, etc. Standard methods for preparing pharmaceutically acceptable salts and formulations thereof are well known in the art and are disclosed in various references, including, for example, "Remington: The Science and Practice of Pharmacy", A. Gennaro, ed., 20th ed., Lippincott, Williams & Wilkins, Philadelphia, PA.

[0070] The terms "carrier" or "vehicle" as used interchangeably herein include carriers, excipients, adjuvants, and diluents or any combination of the foregoing, and refer to a material, composition, or vehicle, such as a liquid or solid filler, diluent, excipient, solvent, or encapsulating material, that is involved in carrying or transporting an agent from one organ or part of the body to another. In addition to adjuvants, excipients, and diluents known to those skilled in the art, carriers also include nanoparticles of both organic and inorganic nature.

[0071] As used herein, the term "treating" with respect to a patient refers to ameliorating at least one symptom of a disease or disorder in the patient. Treatment may be amelioration or at least partial alleviation of the disease or disorder.

[0072] As used herein, the term "patient" or "subject" includes all mammals, and more particularly includes humans. The methods described herein can be used for both human therapy and veterinary applications. In an embodiment, the subject is a human.

[0073] As used herein, "therapeutically effective amount" means an amount of a compound or therapeutic 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 the disease or condition. The therapeutically effective amount will vary depending on the type of compound or therapeutic agent selected, the disease or condition and its severity, and the age, weight, etc., of the patient to be treated.

[0074] As used herein, the term "aliphatic" or "aliphatic group" or "aliphatic chain" means a straight (i.e., unbranched), branched, or cyclic, substituted or unsubstituted hydrocarbon chain that is fully saturated or that contains one or more unsaturated units and has a single point of attachment to the rest of the molecule. Unless otherwise indicated, an aliphatic group contains 1-30 aliphatic carbon atoms. In embodiments, an aliphatic group contains 10-30 aliphatic carbon atoms. In embodiments, an aliphatic group contains 10-20 aliphatic carbon atoms. In embodiments, an aliphatic group contains 15-20 aliphatic carbon atoms. In embodiments, an aliphatic group contains 15-17 aliphatic carbon atoms. In embodiments, an aliphatic group contains 1-10 aliphatic carbon atoms. In embodiments, an aliphatic group contains 1-8 aliphatic carbon atoms. In embodiments, an aliphatic group contains 1-6 aliphatic carbon atoms, and in embodiments, an aliphatic group contains 1-4 aliphatic carbon atoms. An aliphatic group may be a saturated or unsaturated, linear or branched, substituted or unsubstituted alkyl, alkenyl, or alkynyl group.

[0075] As used herein, the term "triglyceride" refers to an ester derived from glycerol and three fatty acids. The fatty acids can be the same or different. The symbols used to describe triglycerides in this specification are the same as those used to describe fatty acids below. The fatty acids can be attached to the glycerol molecule in any order. For example, any fatty acid can react with any hydroxyl group of the glycerol molecule to form an ester bond. For example, in a non-limiting example, a triglyceride can include glycerol with any combination of the following fatty acids: C18:1 fatty acids, C14:1 fatty acids, C16:1 fatty acids, polyunsaturated fatty acids, and saturated fatty acids. A triglyceride of C18:1 fatty acids simply refers to a triglyceride whose fatty acid component is derived from or based on a C18:1 fatty acid. In other words, 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 having 16 carbon atoms, wherein each fatty acid has one double bond. A triglyceride of a C18:1 fatty acid in combination with a C14:1 and / or C16:1 fatty acid refers to: (a) a C18:1 triglyceride mixed with a C14:1 triglyceride or a C16:1 triglyceride or both; or (b) at least one of the fatty acid components of the triglyceride is derived from or based on a C18:1 fatty acid, and the other two are derived from or based on a C14:1 fatty acid and / or a C16:1 fatty acid.

[0076] The term "fatty acid" and similar terms refer to carboxylic acids with long aliphatic tails that are either saturated or unsaturated. The terms "long aliphatic tail" and "fatty acid chain" are used interchangeably herein. As used herein, fatty acid chain lengths include C4 to C30 (e.g., C6 to C30), saturated or unsaturated, cis or trans, Zusammen (Z) or entgegen (E), unsubstituted or substituted with C1-10 side chains.

[0077] Unsaturated fatty acids have more than one double bond between carbon atoms. Saturated fatty acids do not contain any double bonds. In an embodiment, a fatty acid can be described herein by the capital letter "C" representing a carbon atom, followed by a numeral describing the number of carbon atoms in the fatty acid, followed by a colon and another numeral representing the number of double bonds in the fatty acid. For example, C16:1 represents a fatty acid with 16 carbon atoms of a double bond, such as palmitoleic acid. The numeral after the colon in this symbol neither specifies the position of the double bond in the fatty acid nor specifies whether the hydrogen atoms bonded to the carbon atoms of the double bond are cis to each other. Other examples of this symbol include C18:0 (stearic acid), C18:1 (oleic acid), C18:2 (linoleic acid), C18:3 (α-linolenic acid) and C20:4 (arachidonic acid).

[0078] As used herein, term "sterol" refers to an animal or plant steroid containing at least one hydroxyl group. In an embodiment, the sterol of the present disclosure has a single hydroxyl group at the C3-position. Typically, sterol contains 27 to 30 carbon atoms and a double bond in the 5 / 6 position and occasionally in 7 / 8, 8 / 9 or other positions. Non-limiting examples of sterols contemplated herein include sigmasterol, campesterol, sitosterol, stigmasterol, rapeseed sterol, stigmasterol, D5 avenasterol, D7 avenasterol, ergosterol, limonosterol, cholesterol, lanosterol, spongosterol, fungosterol, starfish sterol, yeast sterol, and mixtures thereof. In an embodiment, the sterol is cholesterol.

[0079] As used herein, the term "sterol ester" refers to an ester derived from a sterol as defined above and a carboxylic acid. In an embodiment, the carboxylic acid is of formula R b -C(O)-OH, where R b is an aliphatic group. In embodiments, the carboxylic acid is acetic acid, propionic acid, hexanoic acid, butyric acid, valeric acid, caproic acid, octanoic acid, 2-ethylhexanoic acid, capric acid, cyclopentylpropionic acid, lauric acid, isotridecanoic acid, myristic acid, palmitic acid, palmitoleic acid, stearic acid, isostearic acid, oleic acid, elaidic acid, petroselinic acid, linoleic acid, conjugated linoleic acid (CLA), linolenic acid, elaeosteric acid, arachidic acid, arachidonic acid, gadoleic acid, behenic acid, and erucic acid.

[0080] The term "phospholipid" refers to an amphiphilic compound composed of a glycerol molecule bound to two fatty acid "tails" and a phosphate "head." The phosphate group can be further bound to hydrogen, choline, serine, ethanolamine, or inositol, thus diversifying into phosphatidic acid, phosphatidylcholine, phosphatidylserine, phosphatidylethanolamine, and phosphatidylinositol phospholipids, respectively.

[0081] As used herein, the term "lysophospholipid" refers to a derivative (e.g., as defined above) of a phospholipid in which one of the acyl fatty acid tails has been removed by hydrolysis. Therefore, a lysophospholipid has a free alcohol at the sn-1 or sn-2 position. In non-limiting embodiments, the lysophospholipid is 1-myristoyl-2-hydroxyl-sn-glycero-3-phosphocholine (MHPC), 1-palmitoyl-2-hydroxyl-sn-glycero-3-phosphocholine (PHPC), or 1-stearoyl-2-hydroxyl-sn-glycero-3-phosphocholine (SHPC).

[0082] The terms "apolipoprotein AI" or "apoA-I," and "apolipoprotein Al" or "apoAl" refer to the protein encoded by the APOAI gene in humans. DETAILED DESCRIPTION

[0083] Throughout this disclosure, various patents, patent applications, and publications are cited. The disclosures of these patents, patent applications, and publications are incorporated herein by reference in their entireties for all purposes in order to more fully describe the state of the art as 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, the present disclosure will control.

[0084] Until recently, immunological memory was considered an exclusive feature of the adaptive immune system. However, this view has been challenged by the discovery that innate immune cells also adapt and utilize forms of immunological memory. Following exposure to certain stimuli, monocytes and macrophages can mount faster and more robust inflammatory responses to subsequent, unrelated stimuli.

[0085] Innate immune memory, also known as "trained immunity," is a functional state of metabolically and epigenetically regulated myeloid cells. Trained immunity is important for host defense but can also be modulated to achieve therapeutic goals. For example, suppressing trained immunity can be used to promote organ acceptance after transplantation, while boosting trained immunity can be used to provide anti-cancer effects.

[0086] The epigenetic modifications that underlie trained immunity are tightly coupled to metabolic changes, including the induction of aerobic glycolysis, glutaminolysis, and cholesterol metabolism. Intermediates in these metabolic pathways serve as substrates for epigenetic enzymes or as regulators of these enzymes.

[0087] Nanobiocomposites

[0088] Nanobiologics can contain multiple components, including lipids (eg, phospholipids, sphingolipids, triglycerides, cholesterol) and apolipoprotein A (apoA-1), the major protein component of high-density lipoproteins.

[0089] In an embodiment, the nanobiocomposition is spherical in shape. Spherical nanobiocompositions typically comprise apoA-1 or a peptide mimetic of ApoA-1, one or more phospholipids, one or more sphingolipids, one or more therapeutically active agents (e.g., small molecule drugs or their prodrugs), optionally cholesterol, and a hydrophobic core material such as one or more triglycerides or one or more polymers. For example, the inclusion of one or more triglycerides and / or one or more polymers in the nanoparticles disclosed herein can facilitate the regulation of nanoparticle size (e.g., from about 10 nm to more than 30 nm, or more than 100 nm) and shape (from disc-like to spherical). Conversely, the size, rigidity, and viscosity of the nanobiocomposition can also affect load and biodistribution.

[0090] Without being bound by theory, the inherent propensity of nanobiologics for uptake by bone marrow cells allows for efficient delivery of payloads to the innate immune system.

[0091] In an embodiment, the present disclosure provides a nanobio composition comprising:

[0092] (a) apolipoprotein AI (apoA-I) or a peptide mimetic of apoA-I; and

[0093] (b) Sphingolipids.

[0094] In embodiments, the nanobiocompositions of the present disclosure can further comprise one or more additional components (e.g., 1, 2, 3, or 4 components) independently for each occurrence selected from the group consisting of a phospholipid, a lysophospholipid, a hydrophobic matrix core molecule (one or more), and a sterol (e.g., cholesterol).

[0095] In an embodiment, the present disclosure provides a nanobio composition comprising:

[0096] (a) apolipoprotein AI (apoA-I) or a peptide mimetic of apoA-I;

[0097] (b) sphingolipids; and

[0098] (c) Sterols (eg, cholesterol).

[0099] In an embodiment, the present disclosure provides a nanobio composition comprising:

[0100] (a) apolipoprotein AI (apoA-I) or a peptide mimetic of apoA-I;

[0101] (b) phospholipids; and

[0102] (c) Sphingolipids.

[0103] In an embodiment, the present disclosure provides a nanobio composition comprising:

[0104] (a) apolipoprotein AI (apoA-I) or a peptide mimetic of apoA-I;

[0105] (b) sphingolipids;

[0106] (c) phospholipids; and

[0107] (d) Lysophospholipids.

[0108] In an embodiment, the present disclosure provides a nanobio composition comprising:

[0109] (a) apolipoprotein AI (apoA-I) or a peptide mimetic of apoA-I;

[0110] (b) sphingolipids;

[0111] (c) phospholipids; and

[0112] (d) sterol (e.g., cholesterol). In an embodiment, the present disclosure provides a nanobiocomposition comprising:

[0113] (a) apolipoprotein AI (apoA-I) or a peptide mimetic of apoA-I;

[0114] (b) sphingolipids;

[0115] (c) phospholipids; and

[0116] (d) cholesterol;

[0117] The composition is a nanoparticle having a diameter of about 8 nm to about 150 nm.

[0118] In an embodiment, the present disclosure provides a nanobio composition comprising:

[0119] (a) apolipoprotein AI (apoA-I) or a peptide mimetic of apoA-I;

[0120] (b) sphingolipids;

[0121] (c) phospholipids;

[0122] (d) lysophospholipids; and

[0123] (e) Sterols (eg, cholesterol).

[0124] In an embodiment, the present disclosure provides a nanobio composition comprising:

[0125] (a) apolipoprotein AI (apoA-I) or a peptide mimetic of apoA-I;

[0126] (b) phospholipids;

[0127] (c) cholesterol; and

[0128] (d) sphingolipids;

[0129] wherein the sphingolipids are present as about 1-100 mol % of the total lipid composition of the nanobiocomposition; and

[0130] The composition is a nanoparticle having a diameter of about 8 nm to about 150 nm.

[0131] In an embodiment, the present disclosure provides a nanobio composition comprising:

[0132] (a) apolipoprotein AI (apoA-I) or a peptide mimetic of apoA-I;

[0133] (b) sphingolipids;

[0134] (c) sterols (e.g., cholesterol); and

[0135] (d) A hydrophobic matrix core (eg, triglyceride).

[0136] In an embodiment, the present disclosure provides a nanobio composition comprising:

[0137] (a) apolipoprotein AI (apoA-I) or a peptide mimetic of apoA-I;

[0138] (b) phospholipids;

[0139] (c) sphingolipids; and

[0140] (d) A hydrophobic matrix core (eg, triglyceride).

[0141] In an embodiment, the present disclosure provides a nanobio composition comprising:

[0142] (a) apolipoprotein AI (apoA-I) or a peptide mimetic of apoA-I;

[0143] (b) sphingolipids;

[0144] (c) phospholipids;

[0145] (d) lysophospholipids;

[0146] (e) A hydrophobic matrix core (eg, triglyceride).

[0147] In an embodiment, the present disclosure provides a nanobio composition comprising:

[0148] (a) apolipoprotein AI (apoA-I) or a peptide mimetic of apoA-I;

[0149] (b) sphingolipids;

[0150] (c) phospholipids;

[0151] (d) cholesterol; and

[0152] (e) A hydrophobic matrix core (eg, triglyceride).

[0153] In an embodiment, the present disclosure provides a nanobio composition comprising:

[0154] (a) apolipoprotein AI (apoA-I) or a peptide mimetic of apoA-I;

[0155] (b) sphingolipids;

[0156] (c) phospholipids;

[0157] (d) lysophospholipids;

[0158] (e) sterols (e.g., cholesterol); and

[0159] (f) A hydrophobic matrix core (eg, triglyceride).

[0160] In embodiments of the nanobiocompositions provided herein, the sphingolipid is selected from the group consisting of ceramide, sphingomyelin, dihydroceramide, glucosylceramide, sphingosine, sphingosine-1-phosphate, galactosylceramide, ceramide-1-phosphate, lactosylceramide, and mixtures thereof.

[0161] In embodiments of the nanobiocompositions provided herein, the sphingolipid is selected from the group consisting of dihydroceramide, glucosylceramide, sphingosine, sphingosine-1-phosphate, galactosylceramide, ceramide-1-phosphate, lactosylceramide, and mixtures thereof.

[0162] In embodiments of the nanobiocompositions provided herein, the sphingolipid is selected from the group consisting of dihydroceramide, glucosylceramide, sphingosine, sphingosine-1-phosphate, galactosylceramide, ceramide-1-phosphate, lactosylceramide, and mixtures thereof.

[0163] In the embodiments of the nanobiocompositions provided herein, the sphingolipid is selected from the group consisting of glucosylceramide, galactosylceramide, and lactosylceramide.

[0164] In embodiments of the nanobiocompositions provided herein, the sphingolipid is glucosylceramide or galactosylceramide.

[0165] In embodiments of the nanobiocompositions provided herein, the sphingolipid is a ceramide, in embodiments, the sphingolipid is a sphingomyelin. In embodiments, the sphingolipid is a dihydroceramide. In embodiments, the sphingolipid is a glucosylceramide. In embodiments, the sphingolipid is sphingosine. In embodiments, the sphingolipid is sphingosine-1-phosphate. In embodiments, the sphingolipid is a galactosylceramide. In embodiments, the sphingolipid is ceramide-1-phosphate. In embodiments, the sphingolipid is lactosylceramide.

[0166] In the embodiments of the nanobiocompositions provided herein, the sphingolipid is selected from the group consisting of ceramides, sphingomyelins, cerebrosides, sulfatides, erythrosides, gangliosides, and mixtures thereof.

[0167] In embodiments of the nanobiocompositions provided herein, the sphingolipid is one or more sphingolipids selected from the group consisting of ceramide, phytosphingosine, phosphosphingolipids, glycosphingolipids, or sphingosine. In embodiments, the sphingolipid can be selected from the group consisting of sphingosine, dihydrosphingosine, ceramide, sphingomyelin, gangliosides, glycosphingolipids, phosphosphingolipids, phytosphingosine, and derivatives thereof.

[0168] In embodiments of the nanobiocompositions provided herein, the sphingolipid is one or more sphingolipids selected from the group consisting of phytoceramide, sulfatide, lactosphingolipid, galactosphingolipid, glucosphingolipid, ganglioside, erythroglycoside, 2-hydroxyceramide, dihydroceramide phosphate, dihydroceramide, 1-O-acylceramide, ceramide, methylated sphingosine, phosphorylated sphingosine, dihydrosphingosine. In embodiments, the sphingolipid is a natural sphingosine (e.g., egg sphingosine, brain sphingosine) or a natural ceramide (e.g., ceramide (egg), ceramide (brain), brain CPE).

[0169] In embodiments of the nanobiocompositions provided herein, the sphingolipid is one or more sphingolipids selected from the group consisting of D-ribose-phytosphingosine-1-phosphate, D-ribose-phytosphingosine (C17 base), phytosphingosine-N,N-dimethyl, N-24:0 (2S-OH) phytosphingosine, phytosphingosine-N,N-dimethyl, N-02:0 phytosphingosine, N-08:0 phytosphingosine, N-18:0 phytosphingosine, N-16:0 phytosphingosine, N-24:0 phytosphingosine, N-24:0 (2R-OH) phytosphingosine, 24:0 CPE (d18:1 / 24:0), 24:1 CPE (d18 / 24:1), C12 sphingosine. (Sphingosyl) PE (d17:1 / 12:0), sphingosine PE (d18:1), sphingosine PI (d18:1), KRN7000, sulfatide (brain), C24:1 monothiogalactosyl (β) ceramide (d18:1 / 24:1), C17 monothiogalactosyl (β) ceramide (d18:1 / 17:0), C12 monothiogalactosyl (β) ceramide (d18:1 / 12:0), C12 dithiogalactosyl (β) ceramide (d18:1 / 12:0), C24 monothiogalactosyl (β) ceramide (d18:1 / 24:0), 18:0 (2R-OH) thiogalactosyl ceramide (Sulfo GalCer), 18:0(2S-OH)thiogalactosylceramide, C24:1monothiogalactosyl(α)ceramide (d18:1 / 24:1), C8lactose(β)ceramide (d18:1 / 8:0), lactose(β)sphingosine (d18:1), C12lactose(β)ceramide (d18:1 / 12:0), C8 L-threo-lactose(β)ceramide (d18:1 / 8:0), C16 lactose(β)ceramide (d18:1 / 16:0), C24 lactose(β)ceramide (d18:1 / 24:0), C18:1 lactose(β)ceramide (d18:1 / 18:1), C17 lactose(β)ceramide (d18:1 / 17:0), C24:1 lactose(β)ceramide (d18:1 / 24:1), C18 lactose(β)ceramide (d18:1 / 18:0); C8 galactose(α)ceramide Ceramide (d18:1 / 8:0), C16 galactosyl (α) ceramide (d18:1 / 16:0), C24:1 galactosyl (α) ceramide (d18:1 / 24:1 (15Z)), galactosyl (α) sphingosine (d18:1), adamantyl galactosyl (β) ceramide, C16 galactosyl (β) ceramide (d18:1 / 16:0), galactosyl (β) sphingosine (d18:1), C8 galactosyl (β) ceramide (d18:1 / 8:0), C12 galactosyl (β) ceramide (d18:1 / 12:0),C24:1 galactosyl (β) ceramide (d18:1 / 24:1 (15Z)), galactosyl (β) dimethyl sphingosine (d18:1), C18:1 galactosyl (β) ceramide (d18:1 / 18:1 (9Z)), C16 galactosyl (α) dihydroceramide (d18:0 / 16:0), C16 galactosyl (β) dihydroceramide (d18:0 / 16:0), C18 (2R-OH) galactosyl (β) ceramide, C18 (2S-OH) galactosyl (β) ceramide, C18 galactosyl (β) ceramide (d18:1 / 18:0), C24:0 galactosyl (β) ceramide (d18:1 / 24:0), cerebrosides (brain), glucose ceramide Ceramide (soybean), glucose (β) ceramide (d20:1), glucose (α) ceramide (d18:1), glucose (β) ceramide (d18:1), C16 glucose (β) ceramide (d18:1 / 16:0), C8 glucose (β) ceramide (d18:1 / 8:0), C12 glucose (β) ceramide (d18:1 / 12:0), C18 glucose (β) ceramide (d18:1 / 18:0), C18:1 glucose (β) ceramide (d18:1 / 18:1(9Z)), C24:1 glucose (β) ceramide (d18:1 / 24:1(15Z)), C17 glucose (β) ceramide (d18:1 / 17:0 ), Gangliosides-total, Ganglioside GD1a (porcine brain), Ganglioside GD1b (porcine brain), Ganglioside GM3 (cow milk), Ganglioside GT1b (porcine brain), Ganglioside GD3 (cow milk), NGcGM3 (cow spleen), Ganglioside GM1 (sheep brain), C18:0GM3 (synthetic), C17:0GD1a (d18:1 / 17:0), Ganglioside GQ1b (porcine brain), C20:0GM1 (synthetic), C18:0GT1b (d18:1 / 18:0), C18:0GD1a (d18:1 / 18:0), C17:0GM1 (synthetic), C18:0GM1-biotin (synthetic), C17:0 GB3 (synthetic), C17:0GA2, hemolytic iGB3 (synthetic), C17:0iGB3 (synthetic), hemolytic (lyso)GB3 (synthetic), hemolytic GA2, natural sphingomyelin (egg SM, brain SM, milk SM), synthetic sphingomyelin derivatives (e.g., 02:0SM (d18:1 / 2:0), 06:0SM (d18:1 / 6:0), 12:0SM (d18:1 / 12:0), 16:0SM (d18:1 / 16:0), 17:0SM (d18:1 / 17:0), 18:0SM (d18:1 / 18:0), 18:1SM (d18:1 / 18:1(9Z)), 24:0SM, 24:1SM,LysoSM (d18:1), LysoSM (dihydro) (d18:0), LysoSM-d7, LysoSM (d17:1), 12:0 dihydroSM (d18:0 / 12:0), 16:1SM (d18:1 / 16:1(9Z)), 14:0SM (d18:1 / 14:0), 12:0(2R-OH)ceramide, 2:0(2S-OH)ceramide, 16:0(2R-OH)ceramide, 16:0(2S-OH)ceramide, 17:0(2R-OH)ceramide, 17:0(2S-OH)ceramide, 20 :0(2R-OH)ceramide, 20:0(2S-OH)ceramide, 22:0(2R-OH)ceramide, 22:0(2S-OH)ceramide, 24:0(2R-OH)ceramide, 24:0(2S-OH)ceramide, 24:1(2R-OH)ceramide, 24:1(2S-OH)ceramide, 18:1(2R-OH)ceramide, 18:1(2S-OH)ceramide, 18:0(2R-OH)ceramide, 18:0(2S-OH)ceramide, C16 Dihydroceramide-1-Phosphate (d18:0 / 16:0), C24 Dihydroceramide-1-Phosphate (d24 Dihydroceramide-1-Phosphate)(d18:0 / 24:0), C18:1 dihydroceramide (d18:0 / 18:1(9Z)), C2 dihydroceramide (d18:0 / 2:0), C8 dihydroceramide (d18:0 / 8:0), C18 dihydroceramide (d18:0 / 18:0), C24 dihydroceramide (d18:0 / 24:0), C24:1 dihydroceramide (d18:0 / 24:1(15Z)), C14 dihydroceramide (d18:0 / 14:0), C6 dihydroceramide (d 18:0 / 6:0), 6 dihydroceramide (d18:0 / 16:0), C12 dihydroceramide (d18:0 / 12:0), 1-O-acyl-ceramide, C24 ceramide-1-phosphate (C24Ceramide-1-Phosphate) (d18:1 / 24:0), C2 ceramide-1-phosphate (d18:1 / 2:0), C12 ceramide-1-phosphate (d18:1 / 12:0), C8 ceramide-1-phosphate (d18:1 / 8:0), C16 ceramide-1-phosphate (d18:1 / 16:0), 16:0 3-deoxy-C1P, C18:1 ceramide-1-phosphate (d18:1 / 18:1(9Z)), C8 ceramide-1-phosphate (d17:1 / 8:0), C12 ceramide-1,3-cyclo-phosphate (d18:1 / 12:0),C16 ceramide-1,3-cyclo-phosphate (d18:1 / 16:0), C6 ceramide-1,3-cyclo-phosphate (d18:1 / 6:0), C15 ceramide (d18:1 / 15:0), C22 ceramide (d18:1 / 22:0), C2 ceramide (d18:1 / 2:0), C4 ceramide (d18:1 / 4:0), C6 ceramide (d18:1 / 6:0), C8 ceramide (d18:1 / 8:0), C10 ceramide (d18:1 / 10:0), C12 Ceramide (d18:1 / 12:0), C14 Ceramide (d18:1 / 14:0), C16 Ceramide (d18:1 / 16:0), C17 Ceramide (d18:1 / 17:0), C18 Ceramide (d18:1 / 18:0), C18:1 Ceramide (d18:1 / 18:1(9Z)), C20 Ceramide (d18:1 / 20:0), C24 Ceramide (d18:1 / 24:0), C24:1 Ceramide (d18:1 / 24:1(15Z)), C18 Ceramide (d17:1 / 18:0), C20 Ceramide (d17:1 / 20:0), C24 Ceramide (d17:1 / 24:0), C24:1 Ceramide (d17:1 / 24:1(15Z)), C16 Ceramide (d14:1 / 16:0), C22 Ceramide (d14:1 / 22:0), C24:1 Ceramide (d18:2(4E,8Z) / 24:1(15Z)), C16 :0 ceramide (d18:2(4E,8Z) / 16:0), C24:0 ceramide (d18:2(4E,8Z) / 24:0), ceramide (egg), ceramide (brain), brain CPE, CER1 (d18:1 / 26:0 / 18:1), CER9 (t18:0 / 26:0 / 18:1), monomethyl sphingosine (d18:1), dimethyl sphingosine (d18:1), trimethyl sphingosine (d18:1), dimethyl sphingosine-1-phosphate (Dimethyl Sphingosine-1-Phosphate)(d18:1), Sphingosine-1-Phosphate(d18:1), Dihydrosphingosine-1-Phosphate(d18:0), Sphingosine-1-Phosphate(d17:1), Sphingosine-1-Phosphate(d17:1), Sphingosine-1-Phosphate(d20:1), Dihydrosphingosine-1-Phosphate(d20:0), N-12:0-1-deoxysphingosine, Dihydrosphingosine(d18:0), Dihydrosphingosine(d17:0), Dihydrosphingosine(d20:0), 3-Ketodihydrosphingosine(d18:0), 3-Deoxysphingosine, L-Threo-sphingosine(d18:1), Sphingosine(d18:1),Sphingosine (d17:1), Sphingosine (d20:1), Sphingosine (d22:1), 4E,14Z-Sphingadiene, 4E,8Z-Sphingadiene, 4E,11Z-Sphingadiene, Sphingosine (d16:1), Sphingosine (d14:1), Mito-So, and Sphingosine.

[0170] In an embodiment, the sphingolipid is of formula (I):

[0171]

[0172]

[0173] or a pharmaceutically acceptable salt thereof, wherein:

[0174] R 1 is a saturated aliphatic chain or an unsaturated aliphatic chain;

[0175] R 2 Is H or R 4 , or C(O)R 4 ;

[0176] R 3 Is H, sugar, P(O)(OH)2, or P(O)(OH)O(CH2) n R 5 , where n = 1-10;

[0177] R 4 is a saturated aliphatic chain or an unsaturated aliphatic chain; and

[0178] R 5 It is H, NH2, N(CH3)3 + , OH, or sugar.

[0179] In an embodiment, the sphingolipid is of formula (I):

[0180]

[0181] or a pharmaceutically acceptable salt thereof, wherein:

[0182] R 1 is a saturated aliphatic chain or an unsaturated aliphatic chain;

[0183] R 2 Is H or R 4 , or C(O)R 4 ;

[0184] R 3 Is H, sugar, P(O)(OH)2, or P(O)(OH)O(CH2) n R 5, where n = 1-10;

[0185] R 4 is a saturated aliphatic chain or an unsaturated aliphatic chain; and

[0186] R 5 It is H, NH2, N(CH3)3 + , OH, or sugar.

[0187] In an embodiment of the sphingolipid of formula (I), R 1 It is an aliphatic chain.

[0188] In an embodiment of the sphingolipid of formula (I), R 1 It is C 4-30 Aliphatic chain.

[0189] In an embodiment of the sphingolipid of formula (I), R 1 It is a saturated aliphatic chain.

[0190] In an embodiment of the sphingolipid of formula (I), R 1 It is C 4-30 Saturated aliphatic chain.

[0191] In an embodiment of the sphingolipid of formula (I), R 1 It is C 4-30 Alkyl or C 4-30 Alkenyl.

[0192] In an embodiment of the sphingolipid of formula (I), R 1 It is C 4-30 Alkenyl.

[0193] In an embodiment of the sphingolipid of formula (I), R 1 Is saturated C 15-17 Aliphatic chain or unsaturated C 15-17 Aliphatic chain.

[0194] In an embodiment of the sphingolipid of formula (I), R 1 It is C 15-17 Alkyl or C 15-17 Alkenyl.

[0195] In an embodiment of the sphingolipid of formula (I), R 1 Is saturated C 15-17 Saturated aliphatic chain.

[0196] In an embodiment of the sphingolipid of formula (I), R 1 It is unsaturated C 15-17 Aliphatic chain.

[0197] In an embodiment of the sphingolipid of formula (I), R 1 It is C 15-17 Alkenyl.

[0198] In an embodiment of the sphingolipid of formula (I), R 1 Is saturated C 15 Saturated aliphatic chain.

[0199] In an embodiment of the sphingolipid of formula (I), R 1 It is C 15 alkyl.

[0200] In an embodiment of the sphingolipid of formula (I), R 1 It is unsaturated C 15 Aliphatic chain.

[0201] In an embodiment of the sphingolipid of formula (I), R 1 It is C 15 Alkenyl.

[0202] In an embodiment of the sphingolipid of formula (I), R 1 Is saturated C 17 Saturated aliphatic chain.

[0203] In an embodiment of the sphingolipid of formula (I), R 1 It is C 17 alkyl.

[0204] In an embodiment of the sphingolipid of formula (I), R 1 It is unsaturated C 17 Aliphatic chain.

[0205] In an embodiment of the sphingolipid of formula (I), R 1 It is C 17 Alkenyl.

[0206] In an embodiment, the sphingolipid of formula (I) is of formula (IV-A):

[0207]

[0208] In an embodiment, the sphingolipid of formula (I) is of formula (IV-B):

[0209]

[0210] In embodiments of the sphingolipids of Formula (I), (IV-A), or (IV-B), R 2 It is H, R 4 , or C(O)R 4 In an embodiment, R 2 is H. In an embodiment, R 2 It is R 4 In an embodiment, R 2 It is C(O)R 4 .

[0211] In embodiments of the sphingolipids of Formula (I), (IV-A), or (IV-B), R 3 Is H, sugar, P(O)(OH)2, or P(O)(OH)O(CH2) n R 5 , where n=1-10.

[0212] In embodiments of the sphingolipids of Formula (I), (IV-A), or (IV-B), R 3 It is -H, -P(O)(OH)2 or sugar.

[0213] In embodiments of the sphingolipids of Formula (I), (IV-A), or (IV-B), R 3 Yes -H.

[0214] In embodiments of the sphingolipids of Formula (I), (IV-A), or (IV-B), R 3 It is -P(O)(OH)2.

[0215] In embodiments of the sphingolipids of Formula (I), (IV-A), or (IV-B), R 3 It is P(O)(OH)O(CH2) n R 5 , wherein n=1-10. In embodiments, n is 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10. In embodiments, n is 1. In embodiments, n is 2. In embodiments, n is 3. In embodiments, n is 4. In embodiments, n is 5. In embodiments, n is 6. In embodiments, n is 7. In embodiments, n is 8. In embodiments, n is 9. In embodiments, n is 10.

[0216] In embodiments of the sphingolipids of Formula (I), (IV-A), or (IV-B), R 3 It's sugar.

[0217] In an embodiment, R 3 yes

[0218] In an embodiment, R 3 yes In an embodiment, R 3 yes

[0219]

[0220] In an embodiment, R 3 yes In an embodiment, R 3 yes In an embodiment, R3 yes

[0221] In embodiments of the sphingolipids of Formula (I), (IV-A), or (IV-B), R 5 It is H, NH2, N(CH3)3 + , OH, or sugar.

[0222] In embodiments of the sphingolipids of Formula (I), (IV-A), or (IV-B), R 5 It is H, NH2, OH, or sugar.

[0223] In embodiments of the sphingolipids of Formula (I), (IV-A), or (IV-B), R 5 It’s H.

[0224] In an embodiment, R 5 It's NH2.

[0225] In an embodiment, R 5 It is N(CH3)3 + .

[0226] In an embodiment, R 5 It's OH.

[0227] In an embodiment, R 5 It's sugar.

[0228] In embodiments of the sphingolipid of Formula (I), (IV-A), or (IV-B), the sugar is:

[0229]

[0230] In embodiments of the sphingolipids of Formula (I), (IV-A), or (IV-B), the sugar is In an embodiment, the sugar is:

[0231] In embodiments of the sphingolipid of Formula (I), (IV-A), or (IV-B), the sugar is:

[0232] In embodiments of the sphingolipids of Formula (I), (IV-A), or (IV-B), the sugar is In an embodiment, the sugar is In an embodiment, the sugar is:

[0233] In embodiments of the sphingolipids of Formula (I), (IV-A), or (IV-B), R 4 It is a saturated aliphatic chain or an unsaturated aliphatic chain.

[0234] In embodiments of the sphingolipids of Formula (I), (IV-A), or (IV-B), R 4 It is C 4-30 Saturated aliphatic chain or C 4-30 Unsaturated aliphatic chain.

[0235] In embodiments of the sphingolipids of Formula (I), (IV-A), or (IV-B), R 4 It is C 4-30 Saturated aliphatic chain or C 4-30 Alkenyl.

[0236] In embodiments of the sphingolipids of Formula (I), (IV-A), or (IV-B), R 4 It is C 4-30 Alkyl or C 4-30 Alkenyl.

[0237] In embodiments of the sphingolipids of Formula (I), (IV-A), or (IV-B), R 4 It is C 11-30 Unsaturated aliphatic chain.

[0238] In embodiments of the sphingolipids of Formula (I), (IV-A), or (IV-B), R 4 It is C 11-30 Alkenyl.

[0239] In embodiments of the sphingolipids of Formula (I), (IV-A), or (IV-B), R 4 It is C 11-30 Saturated aliphatic chain.

[0240] In embodiments of the sphingolipids of Formula (I), (IV-A), or (IV-B), R 4 It is C 11-30 alkyl.

[0241] In embodiments of the sphingolipids of Formula (I), (IV-A), or (IV-B), R 4 It is C 11 Unsaturated aliphatic chain.

[0242] In embodiments of the sphingolipids of Formula (I), (IV-A), or (IV-B), R 4 It is C 11 Alkenyl.

[0243] In embodiments of the sphingolipids of Formula (I), (IV-A), or (IV-B), R 4 It is C 15 Unsaturated aliphatic chain.

[0244] In embodiments of the sphingolipids of Formula (I), (IV-A), or (IV-B), R 4 It is C 15 Alkenyl.

[0245] In embodiments of the sphingolipids of Formula (I), (IV-A), or (IV-B), R 4 It is C 17 Unsaturated aliphatic chain.

[0246] In embodiments of the sphingolipids of Formula (I), (IV-A), or (IV-B), R 4 It is C 17 Alkenyl.

[0247] In embodiments of the sphingolipids of Formula (I), (IV-A), or (IV-B), R 4 It is C 23 Unsaturated aliphatic chain.

[0248] In embodiments of the sphingolipids of Formula (I), (IV-A), or (IV-B), R 4 It is C 23 Alkenyl.

[0249] In embodiments of the sphingolipids of Formula (I), (IV-A), or (IV-B), R 4 It is C 11 Saturated aliphatic chain.

[0250] In embodiments of the sphingolipids of Formula (I), (IV-A), or (IV-B), R 4 It is C 11 alkyl.

[0251] In embodiments of the sphingolipids of Formula (I), (IV-A), or (IV-B), R 4 It is C 15 Saturated aliphatic chain.

[0252] In embodiments of the sphingolipids of Formula (I), (IV-A), or (IV-B), R 4 It is C 15 alkyl.

[0253] In embodiments of the sphingolipids of Formula (I), (II), (III), (IV-A), or (IV-B), R 4 It is C 17 Saturated aliphatic chain.

[0254] In embodiments of the sphingolipids of Formula (I), (IV-A), or (IV-B), R 4 It is C 17 alkyl.

[0255] In embodiments of the sphingolipids of Formula (I), (IV-A), or (IV-B), R 4 It is C 23 Saturated aliphatic chain.

[0256] In embodiments of the sphingolipids of Formula (I), (IV-A), or (IV-B), R 4 It is C 23 alkyl.

[0257] In embodiments of the sphingolipids of Formula (I), (IV-A), or (IV-B), R 4 yes

[0258]

[0259] In embodiments of the sphingolipids of Formula (I), (IV-A), or (IV-B), R 4 yes

[0260]

[0261] In embodiments of the sphingolipids of Formula (I), (IV-A), or (IV-B), R 4 yes

[0262]

[0263] In embodiments of the sphingolipids of Formula (I), (IV-A), or (IV-B), R 4 yes

[0264]

[0265] In embodiments of the sphingolipids of Formula (I), (IV-A), or (IV-B), R 4 yes

[0266]

[0267] In embodiments of the sphingolipids of Formula (I), (IV-A), or (IV-B), R 4 yes

[0268]

[0269] In embodiments of the sphingolipids of Formula (I), (IV-A), or (IV-B), R 4 yes

[0270]

[0271] In embodiments of the sphingolipids of Formula (I), (IV-A), or (IV-B), R 4 yes

[0272]

[0273] In embodiments of the sphingolipids of Formula (I), (IV-A), or (IV-B), R 1 and R 4 At least one of them is unsaturated.

[0274] In an embodiment of the sphingolipid of formula (I),

[0275] R 1 It is unsaturated C 14-24 aliphatic chains;

[0276] R 2 It is C(O)R 4 ;

[0277] R 3 is sugar or H; and

[0278] R 4 It is C 14-24 Saturated aliphatic chain or C 14-24 Unsaturated aliphatic chain.

[0279] In an embodiment of the sphingolipid of formula (I),

[0280] R 1 It is unsaturated C 14-24 aliphatic chains;

[0281] R 2 It is C(O)R 4 ;

[0282] R 3 is sugar; and

[0283] R 4 It is C 14-24 Saturated aliphatic chain or C 14-24 Unsaturated aliphatic chain.

[0284] In an embodiment of the sphingolipid of formula (I),

[0285] R 1 It is unsaturated C 14-16 Aliphatic chains (e.g., C 15 alkenyl);

[0286] R 2 It is C(O)R 4 ;

[0287] R 3 is sugar; and

[0288] R 4 It is C17-23 Saturated aliphatic chain or C 17-23 Unsaturated fatty chains (e.g., C 17 Alkenyl, C 23 Alkyl or C 23 alkenyl).

[0289] In an embodiment of the sphingolipid of formula (I),

[0290] R 1 It is unsaturated C 14-24 aliphatic chains;

[0291] R 2 It is C(O)R 4 ;

[0292] R 3 is sugar; and

[0293] R 4 It is C 14-24 Unsaturated aliphatic chain.

[0294] In an embodiment of the sphingolipid of formula (I),

[0295] R 1 It is unsaturated C 14-16 Aliphatic chains (e.g., C 15 alkenyl);

[0296] R 2 It is C(O)R 4 ;

[0297] R 3 is sugar; and

[0298] R 4 It is C 17-23 Unsaturated fatty chains (e.g., C 17 Alkenyl, C 23 Alkyl or C 23 In an embodiment, the sphingolipid of formula (I) is a sphingolipid of formula (II):

[0299]

[0300] in

[0301] R 1 It is unsaturated C 14-24 aliphatic chains;

[0302] R 2 It is C(O)R 4 ;and

[0303] R 4 It is C 14-24 Unsaturated aliphatic chain.

[0304] In an embodiment, the sphingolipid of formula (II) is a sphingolipid of formula (II-A) or (II-B):

[0305]

[0306] in

[0307] R 1 It is unsaturated C 14-24 aliphatic chains;

[0308] R 2 It is C(O)R 4 ;and

[0309] R 4 It is C 14-24 Unsaturated aliphatic chain.

[0310] In an embodiment, the sphingolipid of formula (II) is a sphingolipid of formula (II-A):

[0311] In an embodiment, the sphingolipid of formula (II) is a sphingolipid of formula (II-B):

[0312]

[0313] In embodiments of the compounds of Formula (II), (II-A), or (II-B), R 1 It is unsaturated C 15-17 In an embodiment, R 1 It is C 15-17 In an embodiment, R 1 It is C 15 Alkenyl.

[0314] In embodiments of the compounds of Formula (II), (II-A), or (II-B), R 4 It is C 17-23 In an embodiment, R 4 It is C 17 Unsaturated fatty chain, or C 23 In an embodiment, R 4 It is C 17-23 In an embodiment, R 4 It is C 17 Alkenyl or C 23 Alkenyl.

[0315] In embodiments of the nanobio compositions provided herein, the sphingolipid is one or more sphingolipids selected from Table 1.

[0316] In the embodiments of the nanobiocompositions provided herein, the sphingolipid is one or more sphingolipids selected from Table 1, or a pharmaceutically acceptable salt thereof.

[0317] Table 1.

[0318]

[0319]

[0320]

[0321]

[0322]

[0323] In an embodiment, the sphingolipid is selected from the group consisting of:

[0324] (C16 ceramide (d18:1 / 16:0));

[0325]

[0326] (C24:1 galactosyl(β)ceramide (d18:1 / 24:1));

[0327]

[0328] (24:0 sphingomyelin);

[0329]

[0330] (C16 ceramide (d18:1 / 16:0));

[0331]

[0332] (C24:1 galactosyl(β)ceramide (d18:1 / 24:1));

[0333]

[0334] (C24:0 lactose (β) ceramide (d18:1 / 24:0)); and

[0335]

[0336] (C18:1 glucosyl(β)ceramide (d18:1 / 18:1(9Z))).

[0337] In an embodiment, the sphingolipid is selected from the group consisting of:

[0338] (sphingosine (d20:1));

[0339]

[0340] (C24 ceramide (d18:1 / 24:0));

[0341]

[0342] (C16 dihydroceramide (18:0 / 16:0));

[0343]

[0344] (C12 ceramide-1-phosphate (d18:1 / 12:0));

[0345]

[0346] (C16 galactosyl(β)ceramide (d18:1 / 16:0));

[0347]

[0348] ((C24:0 galactosyl(β)ceramide (d18:1 / 24:0));

[0349]

[0350] ((C24:1 lactose (β) ceramide (d18:1 / 24:1));

[0351]

[0352] ((24:0 sphingomyelin));

[0353]

[0354] ((C18:0 glucosyl(β)ceramide (d18:1 / 18:0)); and

[0355]

[0356] C16 ceramide-1-phosphate (d18:1 / 16:0).

[0357] In the embodiments of the nanobiocompositions provided herein, the sphingolipid is:

[0358]

[0359] Sphingosine-1-phosphate (d18:1 / 16:0)

[0360] In the embodiments of the nanobiocompositions provided herein, the sphingolipid is:

[0361]

[0362] (C16 Ceramide (d18:1 / 16:0)).

[0363] In the embodiments of the nanobiocompositions provided herein, the sphingolipid is:

[0364]

[0365] (Sphingosine (d20:1)).

[0366] In the embodiments of the nanobiocompositions provided herein, the sphingolipid is:

[0367]

[0368] (C24 ceramide (d18:1 / 24:0)).

[0369] In the embodiments of the nanobiocompositions provided herein, the sphingolipid is

[0370]

[0371] (C16 dihydroceramide (18:0 / 16:0)).

[0372] In the embodiments of the nanobiocompositions provided herein, the sphingolipid is

[0373]

[0374] C24 dihydroceramide (d18:0 / 24:0).

[0375] In the embodiments of the nanobiocompositions provided herein, the sphingolipid is

[0376] C16 ceramide-1-phosphate (d18:1 / 16:0).

[0377] In the embodiments of the nanobiocompositions provided herein, the sphingolipid is

[0378]

[0379] C18:1 ceramide-1-phosphate (d18:1 / 18:1(9Z)).

[0380] In the embodiments of the nanobiocompositions provided herein, the sphingolipid is

[0381]

[0382] C12 ceramide-1-phosphate (d18:1 / 12:0).

[0383] In the embodiments of the nanobiocompositions provided herein, the sphingolipid is

[0384]

[0385] C16 galactosyl(α)ceramide (d18:1 / 16:0).

[0386] In the embodiments of the nanobiocompositions provided herein, the sphingolipid is

[0387]

[0388] C16 galactosyl(β)ceramide (d18:1 / 16:0).

[0389] In the embodiments of the nanobiocompositions provided herein, the sphingolipid is

[0390] C24:0 galactosyl(β)ceramide (d18:1 / 24:0).

[0391] In the embodiments of the nanobiocompositions provided herein, the sphingolipid is

[0392]

[0393] C24:1 galactosyl(β)ceramide (d18:1 / 24:1).

[0394] In the embodiments of the nanobiocompositions provided herein, the sphingolipid is

[0395]

[0396] C24:0 lactose (β) ceramide (d18:1 / 24:0).

[0397] In the embodiments of the nanobiocompositions provided herein, the sphingolipid is

[0398]

[0399] C24:1 lactose (β) ceramide (d18:1 / 24:1).

[0400] In the embodiments of the nanobiocompositions provided herein, the sphingolipid is

[0401]

[0402] 24:0 sphingomyelin.

[0403] In the embodiments of the nanobiocompositions provided herein, the sphingolipid is

[0404]

[0405] 24:1 sphingomyelin.

[0406] In the embodiments of the nanobiocompositions provided herein, the sphingolipid is

[0407] C18:0 glucosyl(β)ceramide (d18:1 / 18:0).

[0408] In the embodiments of the nanobiocompositions provided herein, the sphingolipid is

[0409]

[0410] C18:1 glucosyl(β)ceramide (d18:1 / 18:1(9Z)).

[0411] In embodiments of the nanobiocompositions provided herein, sphingolipids are present at about, or at least about 1-100 mol% of the total lipid composition of the nanobiocomposition, 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%, about or at least about 30 mol%, about or at least about 31 mol%, about or at least about 32 mol%, about or at least about 33 mol%, about or at least about 34 mol%, about or at least about 35 mol%, about or at least about 36 mol%, about or at least about 37 mol%, about or at least about 38 mol%, about or at least about %, about or at least about 50 mol%, about or at least about 55 mol%, about or at least about 60 mol%, about or at least about 65%, about or at least about 70%, about or at least about 75%, about or at least about 80%, about or at least about 85%, about or at least about 90%, about or at least about 95% to about 100%, including all values ​​and subranges therebetween.

[0412] In the embodiments of the nanobiocompositions provided herein, sphingolipids are present as about 10-25 mol% of the total lipid composition of the nanobiocomposition.

[0413] In the embodiments of the nanobiocompositions provided herein, the sphingolipids are present as about 20 mol% of the total lipid composition of the nanobiocomposition.

[0414] In an embodiment, the nanobiocomposition comprises human apolipoprotein AI (apoA-I); for example, the ApoA-I sequence can have the mature peptide sequence shown below, or can include either or both of a signal peptide (e.g., SEQ ID NO: 347) or a propeptide portion (e.g., SEQ ID NO: 348). The peptide can be produced by expressing a nucleic acid encoding all three of the signal peptide, propeptide portion, and mature peptide. During protein synthesis and maturation, the mature peptide is released and can be purified from the host cell for use in the compositions herein. In certain host cells, such as bacteria, the encoded peptide may not be cleaved into the mature peptide. The N-terminal methionine (M) residue of the Apo-A1 peptide used herein can be a formylmethionine (fM) residue. In an embodiment, the ApoA-I used in the compositions herein comprises or consists of the sequence of SEQ ID NO: 349 or a sequence having 1, 2, 4, 5, 6, 7, 8, 9, 10, or up to 20 conservative substitutions therein.

[0415] Table 2A. ApoA-I peptides

[0416]

[0417] In an embodiment, the nanobiocomposition comprises a peptide mimetic of apolipoprotein AI (apoA-I). Suitable apoA-I mimetic polypeptides may have the sequences shown in Table 2B (SEQ ID NOS: 256 to 263, and 342 to 346) or SEQ ID NOS: 1 to 341.

[0418] Table 2B. ApoA-I mimetics

[0419] SEQ ID NO Amino acid sequence 256 DWLKAFYDKVAEKLKEAF(18A) 257 <![CDATA[Ac-DWLKAFYDKVAEKLKEAF-NH2(2F)]]> 260 <![CDATA[Ac-DWFKAFYDKVAEKFKEAF-NH2(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-NH2(4F-R) q ]]> 345 <![CDATA[Ac-DWFKAFYDRVAERFKEAF-NH2(4F-R’) q ]]> 346 <![CDATA[Ac-DWLXAFYDXVAEXLXEAF-NH2(2F’)]]>

[0420] In an embodiment, the apoA-I mimetic is DWLKAFYDKVAEKLKEAF (SEQ ID NO. 256). In an embodiment, the apoA-I mimetic is Ac-DWLKAFYDKVAEKLKEAF-NH2 (SEQ ID NO. 257). In an embodiment, the apoA-I mimetic is Ac-DWFKAFYDKVAEKFKEAF-NH2 (SEQ ID NO. 260).

[0421] In embodiments, the apoA-I mimetic is optionally acetylated at the N-terminus, or optionally amidated at the C-terminus. In embodiments, the apoA-I mimetic is acetylated at the N-terminus. In embodiments, the apoA-I mimetic is amidated at the C-terminus. In embodiments, the apoA-I mimetic is acetylated at the N-terminus and amidated at the C-terminus.

[0422] In embodiments, the nanobiocompositions of the present disclosure comprise more than one phospholipid. Examples of suitable phospholipids include, but are not limited to, phosphatidylcholine, phosphatidylethanolamine, phosphatidylinositol, phosphatidylserine, and phospholipid-containing oils such as lecithin oil. Non-limiting examples of phospholipids that can be used in the compositions of the present invention include phosphatidylcholine (PC), phosphatidylglycerol (PG), phosphatidylserine (PS), phosphatidylethanolamine (PE), and phosphatidic acid / ester (PA).

[0423] In an embodiment, the phospholipids are independently selected from the group consisting of phosphatidylcholine, phosphatidylethanolamine, phosphatidylinositol, phosphatidylserine, phospholipid-containing oils, phosphatidylglycerol, phosphatidic acid, and combinations thereof.

[0424] In an embodiment, the phospholipid is one or more of the following: DDPC CAS-3436-44-0 1,2-didecanoyl-sn-glycero-3-phosphocholine, DEPA-NA CAS-80724-31-8 1,2-dierucoyl-sn-glycero-3-phosphate (sodium salt), DEPC CAS-56649-39-9 1,2-dierucoyl-sn-glycero-3-phosphocholine, DEPE CAS-988-07-2 1,2-dierucoyl-sn-glycero-3-phosphoethanolamine, DEPG-NA 1,2-dierucoyl-sn-glycero-3-phospho-rac-(l-glycerol) (sodium salt), DLOPC CAS-998-06-1, 1,2-dilinoleoyl-sn-glycero-3-phosphocholine, DLPA-NA 1,2-dilauroyl-sn-glycero-3-phosphate (sodium salt), DLPC CAS-18194-25-7 1,2-Dilauroyl-sn-glycero-3-phosphocholine, DLPE 1,2-Dilauroyl-sn-glycero-3-phosphoethanolamine, DLPG-NA l,2-Dilauroyl-sn-glycero-3-phospho-rac-(l-glycerol) (sodium salt), DLPG-NH4 1,2-Dilauroyl-sn-glycero-3-phospho-rac-(l-glycerol) (ammonium salt), DLPS-NA 1,2-Dilauroyl-sn-glycero-3-phosphoserine (sodium salt), DMPA-NA CAS-80724-3 1,2-Dimyristoyl-sn-glycero-3-phosphate (sodium salt), DMPC CAS-18194-24-6 1,2-Dimyristoyl-sn-glycero-3-phosphocholine, DMPE CAS-988-07-2 1,2-Dimyristoyl-sn-glycero-3-phosphoethanolamine, DMPG-NA CAS-67232-80-8 1,2-Dimyristoyl-sn-glycero-3-phospho-rac-(l-glycerol) (sodium salt), DMPG-NH4 1,2-Dimyristoyl-sn-glycero-3-phospho-rac-(l-glycerol) (ammonium salt), DMPG-NH4 / NA 1,2-Dimyristoyl-sn-glycero-3-phospho-rac-(l-glycerol) (sodium salt / ammonium salt), DMPS-NA 1,2-Dimyristoyl-sn-glycero-3-phospho-rac-(l-glycerol) (sodium salt), DOPA-NA 1,2-Dioleoyl-sn-glycero-3-phosphate (sodium salt), DOPC CAS-4235-95-4 1,2-Dioleoyl-sn-glycero-3-phosphocholine, DOPECAS-4004-5-1 1,2-Dioleoyl-sn-glycero-3-phosphoethanolamine, DOPG-NA CAS-62700-69-0 1,2-Dioleoyl-sn-glycero-3-phospho-rac-(l-glycerol) (sodium salt), DOPS-NA CAS-70614-14-1, 1,2-Dioleoyl-sn-glycero-3-phosphoserine (sodium salt), DPPA-NA CAS-71065-87-7, 1,2-Dipalmitoyl-sn-glycero-3-phosphate (sodium salt), DPPC CAS-63-89-8, 1,2-Dipalmitoyl-sn-glycero-3-phosphocholine, DPPE CAS-923-61-5, 1,2-Dipalmitoyl-sn-glycero-3-phosphoethanolamine, DPPG-NA CAS-67232-81-9, 1,2-Dipalmitoyl-sn-glycero-3-phospho-rac-(l-glycerol) (sodium salt), DPPG-NH4 CAS-73548-70-6 1,2-Dipalmitoyl-sn-glycero-3-phospho-rac-(l-glycerol) (ammonium salt), DPPS-NA l,2-Dipalmitoyl-sn-glycero-3-phosphoserine (sodium salt), DSPA-NA CAS-108321-18-2 1,2-Distearoyl-sn-glycero-3-phosphocholine (sodium salt), DSPC CAS-816-94-4 1,2-Distearoyl-sn-glycero-3-phosphocholine, DSPE CAS-1069-79-0 1,2-Distearoyl-sn-glycero-3-phosphoethanolamine, DSPG-NA CAS-67232-82-0 1,2-Distearoyl-sn-glycero-3-phospho-rac-(l-glycerol) (sodium salt), DSPG-NH4 CAS-108347-80-4 1,2-Distearoyl-sn-glycero-3-phospho-rac-(l-glycerol) (ammonium salt), DSPS-NA 1,2-Distearoyl-sn-glycero-3-phosphoserine (sodium salt), EPC egg-PC, HEPC hydrogenated egg-PC, HSPC hydrogenated soybean PC, MPPC l-myristoyl-2-palmitoyl-sn-glycero-3-phosphocholine, MSPC l-myristoyl-2-stearoyl-sn-glycero-3-phosphocholine, PMPC l-palmitoyl-2-myristoyl-sn-glycero-3-phosphocholine, POPC CAS-26853-31-6l-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine, POPE l-palmitoyl-2-oleoyl-sn-glycero-3-phosphoethanolamine, POPG-NACAS-81490-05-3l-palmitoyl-2-oleoyl-sn-glycero-3[phospho-rac-(l-glycerol)] (sodium salt), PSPC 1-palmitoyl-2-stearoyl-sn-glycero-3-phosphocholine, SMPC l-stearoyl-2-myristoyl-sn-glycero-3-phosphocholine, SOPC l-stearoyl-2-oleoyl-sn-glycero-3-phosphocholine, SPPC1-stearoyl 1-2-palmitoyl l-sn-glycero-3-phosphocholine.

[0425] In an embodiment, the phospholipid is dimyristoylphosphatidylcholine (DMPC), soy lecithin, dipalmitoylphosphatidylcholine (DPPC), distearoylphosphatidylcholine (DSPC), dilauroylphosphatidylcholine (DLPC), dioleoylphosphatidylcholine (DOPC), dilauroylphosphatidylglycerol (DLPG), dimyristoylphosphatidylglycerol (DMPG), dipalmitoylphosphatidylglycerol (DPPG), distearoylphosphatidylglycerol (DSPG), dioleoylphosphatidylglycerol (DOPG), dimyristoylphosphatidic acid (DMPA), dimyristoylphosphatidic acid (DMPA), dipalmitoylphosphatidic acid (DPPA), dipalmitoylphosphatidic acid (DPPA), dimyristoylphosphatidylethanolamine (DMPE), dipalmitoylphosphatidylethanolamine (DPPE), dimyristoylphosphatidylserine (DMPS), dipalmitoylphosphatidylserine (DPPS), and mixtures thereof.

[0426] In embodiments of the nanobiocompositions provided herein, the phospholipid is selected from the group consisting of 1,2-dimyristoyl-sn-glycero-3-phosphocholine (DMPC), 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC), 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC), and mixtures thereof.

[0427] In embodiments of the nanobiocompositions provided herein, the phospholipid is 1,2-dimyristoyl-sn-glycero-3-phosphocholine (DMPC) or 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC).

[0428] In embodiments of the nanobiocompositions provided herein, the phospholipid is DMPC.

[0429] In an embodiment, the nanobiocomposition comprises a phospholipid and a lysophospholipid.

[0430] In an embodiment, the lysophospholipid is lysophosphatidylcholine.

[0431] In embodiments, the lysophospholipid is LYSOPC MYRISTIC 1-myristoyl-sn-glycero-3-phosphocholine, LYSOPC PALMITIC CAS-17364-16-81-palmitoyl-sn-glycero-3-phosphocholine, or LYSOPC STEARIC CAS-19420-57-61-stearoyl-sn-glycero-3-phosphocholine.

[0432] In embodiments of the nanobiocompositions provided herein, the lysophospholipid is selected from the group consisting of 1-myristoyl-2-hydroxy-sn-glycero-3-phosphocholine (MHPC), 1-palmitoyl-2-hydroxy-sn-glycero-3-phosphocholine (PHPC), 1-stearoyl-2-hydroxy-sn-glycero-3-phosphocholine (SHPC), and mixtures thereof.

[0433] In embodiments of the nanobiocompositions provided herein, the phospholipid is 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC) and the lysophospholipid is 1-palmitoyl-2-hydroxy-sn-glycero-3-phosphocholine (PHPC).

[0434] In embodiments, when the composition comprises two types of phospholipids, 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 therebetween.

[0435] In an embodiment, when the composition comprises a phospholipid and a lysophospholipid, the weight ratio of the two phospholipids to the lysophospholipid 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 therebetween.

[0436] In embodiments, the nanobiocompositions of the present disclosure comprise a sterol, such as cholesterol. Typically, the nanobiocompositions comprise from about 1 mol% to about 100 mol% cholesterol relative to phospholipids, 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%, about 22 mol%, about 23 mol%, about 24 mol%, about 25 mol%, about 26 mol%, about 27 mol%, about 28 mol%, about 29 mol%, about 30 mol%, about 31 mol%, about 32 mol%, about 33 mol%, about 34 mol%, about 35 mol%, about 36 mol%, about 37 mol%, about 38 mol%, about 39 mol%, about 40 mol%, about 41 mol%, about 42 mol%, about 43 mol%, about 44 mol%, about 45 mol%, about 46 mol%, about 47 mol%, about 48 mol%, about 49 mol%, about 50 mol%, about 51 mol%, about 52 mol%, about 53 mol%, about 54 mol%, about 55 mol%, about 56 mol%, about 57 mol%, about 58 mol%, about 59 mol%, about 60 mol%, about 61 mol%, about 62 mol%, about 63 mol%, about 64 mol%, about 65 mol%, about 66 %, about 24 mol%, about 25 mol%, about 26 mol%, about 27 mol%, about 28 mol%, about 29 mol%, about 30 mol%, about 35 mol%, about 40 mol%, about 45 mol%, about 50 mol%, about 55 mol%, about 60 mol%, about 65 mol%, about 70 mol%, about 75 mol%, about 80 mol%, about 85 mol%, about 90 mol%, about 95 mol%, to about 100 mol% (i.e., a 1:1 mol / mol mixture of cholesterol and phospholipid (e.g., DMPC)), including all ranges and values ​​therebetween. In embodiments, the nanobiocomposition comprises from about 1 mol% to about 30 mol% cholesterol. In embodiments, the nanobiocomposition comprises from about 15 mol% to about 25 mol% cholesterol relative to phospholipids. In embodiments, the nanobiocomposition comprises from about 20 mol% cholesterol relative to phospholipids. In embodiments, the nanobiocomposition comprises from about 10 mol% to about 35 mol% cholesterol relative to phospholipids. In embodiments, the nanobiocomposition comprises from about 15 mol% to about 30 mol% cholesterol relative to phospholipids. In embodiments, the nanobiocomposition comprises from about 15 mol% to about 25 mol% cholesterol relative to phospholipids. In embodiments, the nanobiocomposition comprises from about 28 mol% to about 23 mol% cholesterol relative to phospholipids. In embodiments, the nanobiocomposition comprises from about 20 mol% to about 27 mol% cholesterol relative to phospholipids.

[0437] Without being bound by theory, the addition of cholesterol may stabilize the nanobiocomposition and improve encapsulation efficiency.

[0438] In embodiments of the nanobio compositions provided herein, cholesterol is present at about 1-30 mol% relative to phospholipids.

[0439] In embodiments of the nanobio compositions provided herein, cholesterol is present at about 5-25 mol% relative to phospholipids.

[0440] In the embodiments of the nanobio compositions provided herein, the nanobio compositions do not contain cholesterol.

[0441] In the embodiments of the nanobiocompositions provided herein, the molar ratio of cholesterol:phospholipid in the nanobiocomposition is 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, or about 0.5:1, including all values ​​therebetween. In embodiments, the cholesterol:phospholipid molar ratio 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, including all ranges therebetween. In embodiments, the cholesterol:phospholipid molar ratio ranges from about 0.05:1 to about 0.25:1. In embodiments, the cholesterol molar ratio is about 0.2:1.

[0442] In embodiments, the weight percentage of cholesterol ranges from about 0% (w / w) to about 15% (w / w) of the lipid or nanobio composition, including from about 1% (w / w), about 1.5% (w / w), about 2% (w / w), about 2.5% (w / w), about 3% (w / w), about 3.5% (w / w), about 4% (w / w), about 4.5% (w / w), about 5% (w / w), about 5.5% (w / w), about 6% (w / w), about 6.5% (w / w), about 7% (w / w), about 8% (w / w), about 9% (w / w), about 10% (w / w), about 11% (w / w), about 12% (w / w), about 13% (w / w), about 14% (w / w), about 15% (w / w), about 16% (w / w), about 17% (w / w), about 18% (w / w), about 19% (w / w), about 20% (w / w), about 21% (w / w), about 22% (w / w), about 23% (w / w), about 24% (w / w), about 25% (w / w), about 26% (w / w), about 27% (w / w), about 28% (w / w), about 29% (w / w), about 30% (w / w), about 31% (w / w), about 32% (w / w), about 33% (w / w), about 35% (w / w), about 36% (w / w), about 37% (w / w), about 38% (w / w), about 39% (w / w), about 41% (w / w), about 42% (w / w), about 43% (w / w), about 4 / w), about 7.5% (w / w), about 8% (w / w), about 8.5% (w / w), about 9% (w / w), about 9.5% (w / w), about 10% (w / w), about 10.5% (w / w), about 11% (w / w / ), about 11.5% (w / w), about 12% (w / w), about 12.5% ​​(w / w), about 13% (w / w), about 13.5% (w / w), about 14% (w / w), about 14.5% (w / w), to about 15% (w / w). In embodiments, the weight percentage of cholesterol ranges from about 0% (w / w) to about 15% of the nanoparticle, lipid, or composition, including from about 1% (w / w), about 1.5% (w / w), about 2% (w / w), about 2.5% (w / w), about 3% (w / w), about 3.5% (w / w), about 4% (w / w), about 4.5% (w / w), about 5% (w / w), about 5.5% (w / w), about 6% (w / w), about 6.5% (w / w), about 7% (w / w), about 8% (w / w), about 9% (w / w), about 10% (w / w), about 11% (w / w), about 12% (w / w), about 13% (w / w), about 14% (w / w), about 15% (w / w), about 16% (w / w), about 17% (w / w), about 18% (w / w), about 19% (w / w), about 20% (w / w), about 21% (w / w), about 22% (w / w), about 23% (w / w), about 24% (w / w), about 25% (w / w), about 26% (w / w), about 27% (w / w), about 28% (w / w), about 29% (w / w), about 30% (w / w), about 31% (w / w), about 32% (w / w), about 33% (w / w), about 35% (w / w), about 36% (w / w), about 37% (w / w), about 38% (w / w), about 39% (w / w), about 40% (w / w), about 41% (w / w), about In some embodiments, the weight percentage is the weight percentage of cholesterol relative to phospholipids. In some embodiments, the weight percentage is the weight percentage of cholesterol relative to total lipids. In some embodiments, the weight percentage is the weight percentage of cholesterol relative to the weight of the nanobio composition. In some embodiments, the weight percentage of cholesterol ranges from about 1 to 10% cholesterol (w / w %) of the nanobio composition. The weight percent of cholesterol ranges from about 2 to 8% cholesterol (w / w %) of the nanobiocomposition. In embodiments, the weight percent of cholesterol ranges from about 3.5 to 7.5% cholesterol (w / w %) of the nanobiocomposition. In embodiments, the weight percent of cholesterol ranges from about 5 to 10% cholesterol (w / w %) of the nanobiocomposition. In embodiments, the weight percent of cholesterol is about 3.6 (w / w %) of the nanobiocomposition.In embodiments, the weight percent of cholesterol is about 7.2 (w / w %) of the nanobio composition. In embodiments, the weight percent of cholesterol is about 5.9 (w / w %) of the nanobio composition.

[0443] In embodiments, the weight percentage of cholesterol in the nanobio compositions of the present disclosure ranges from about 0% (w / w) to about 15% (w / w) of cholesterol relative to phospholipids, including from about 1% (w / w), about 1.5% (w / w), about 2% (w / w), about 2.5% (w / w), about 3% (w / w), about 3.5% (w / w), about 4% (w / w), about 4.5% (w / w), about 5% (w / w), about 5.5% (w / w), about 6% (w / w), about 6.5% (w / w), about 7% (w / w), about 8% (w / w), about 9% (w / w), about 10% (w / w), about 11% (w / w), about 12% (w / w), about 13% (w / w), about 14% (w / w), about 15% (w / w), about 16% (w / w), about 17% (w / w), about 18% (w / w), about 19% (w / w), about 20% (w / w), about 21% (w / w), about 22% (w / w), about 23% (w / w), about 24% (w / w), about 25% (w / w), about 26% (w / w), about 27% (w / w), about 28% (w / w), about 29% (w / w), about 30% (w / w), about 31% (w / w), about 32% (w / w), about 33% (w / w), about 34% (w / w), about 35% (w / w), about 36% (w / w), about 37% (w / w), about 38% (w / w), about 39% (w / w), about 41% (w / w), about About 7% (w / w), about 7.5% (w / w), about 8% (w / w), about 8.5% (w / w), about 9% (w / w), about 9.5% (w / w), about 10% (w / w), about 10.5% (w / w), about 11% (w / w / ), about 11.5% (w / w), about 12% (w / w), about 12.5% ​​(w / w), about 13% (w / w), about 13.5% (w / w), about 14% (w / w), about 14.5% (w / w), and about 15% (w / w).

[0444] In embodiments, the size and circulation time of the nanoparticles can be adjusted, for example, by controlling the ratio of lipid to ApoA-I and the ratio of lipid to polymer or lipid to triglyceride.

[0445] In embodiments, the nanobiocomposition comprises a ratio (e.g., on a molar basis) of phospholipids and / or sphingolipids:ApoA-I or apoA-I mimetic from about 5:1 to 1000:1, including about 5:1, about 10:1, about 20:1, about 30:1, about 40:1, about 50:1, about 60:1, about 70:1, about 80:1, about 90:1, about 100:1, about 110:1, about 120:1, about 130:1, about 140:1, about 150:1, about 160:1, about 170:1, about 180:1, about 190:1, about 200:1, about 210:1, about 220:1, about 230:1, about 240:1, about 250:1, about 260:1, about 270:1, about 280:1, about 290:1, about 300:1, about 310:1, about 320:1, about 330:1, about 340:1, about 350:1, about 360:1, about 370:1, about 380:1, about 390:1, about 400:1 :1, about 200:1, about 210:1, about 220:1, about 230:1, about 240:1, about 250:1, about 260:1, about 270:1, about 280:1, about 290:1, about 300:1, about 310:1, about 320:1, about 330:1, about 340:1, about 350:1, about 360:1, about 370:1, about 380:1, about 390:1, about 400:1, about 410:1, about 420:1, about 430:1, about 440:1, about 450:1, about 460:1, about 470:1 0:1, about 480:1, about 490:1, about 500:1, about 510:1, about 520:1, about 530:1, about 540:1, about 550:1, about 560:1, about 570:1, about 580:1, about 590:1, about 600:1, about 610:1, about 620:1, about 630:1, about 640:1, about 650:1, about 660:1, about 670:1, about 680:1, about 690:1, about 700:1, about 710:1, about 720:1, about 730:1, about 740:1, about 7 : 1, about 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, and to about 1000:1, including all subranges and values ​​therebetween.

[0446] In embodiments, nanobio compositions comprise a lipid:apoA-I ratio (e.g., on a molar basis) of from about 70:1 to 125:1. In embodiments, HDL-derived nanoparticles comprise a lipid:apoA-I mimetic at a ratio (e.g., on a molar basis) of from about 5:1 to 10:1.

[0447] In embodiments, the HDL-derived nanoparticles comprise from about a 2:1 to 3:1 weight ratio of lipid:apoA-I or apoA-I mimetic.

[0448] In embodiments of the nanobio compositions provided herein, the phospholipids, sphingolipids, and cholesterol are present in a molar ratio of about 1:0.05-0.25:0.05-0.25.

[0449] In embodiments of the nanobio compositions provided herein, the phospholipids, sphingolipids, and cholesterol are present in a molar ratio of about 1:0.15-0.25:0.1-0.25.

[0450] In embodiments of the nanobio compositions provided herein, phospholipids, sphingolipids, and cholesterol are present in a molar ratio of about 1:0.2:0.1-0.3.

[0451] In embodiments of the nanobio compositions provided herein, the phospholipids, phospholipids, sphingolipids, and cholesterol are present in a molar ratio of about 1:0.2:0.2.

[0452] In an embodiment, the nanobiocomposition comprises i) apoA-I or a peptide mimetic of apoA-I; ii) a phospholipid; iii) a lysophospholipid, and iv) cholesterol.

[0453] In an embodiment, a nanobio composition comprises i) apoA-I or a peptide mimetic of apoA-I; ii) a phospholipid; and iii) cholesterol.

[0454] In an embodiment, the nanobiocomposition comprises i) apoA-I or a peptide mimetic of apoA-I; ii) a phospholipid; iii) a lysophospholipid, iv) a hydrophobic matrix core, and v) cholesterol.

[0455] In embodiments, the structure and properties (e.g., particle size, rigidity, viscosity, loading, etc.) of HDL-derived nanoparticles can be modified by incorporating a hydrophobic matrix. As used herein, a hydrophobic matrix refers to the core or filler or structural modifier of a nanobiological formulation. Non-limiting examples of suitable hydrophobic matrix molecules include triglycerides, fatty acid esters, hydrophobic polymers, sterol esters, or combinations thereof.

[0456] Any suitable synthetic or natural fatty acid or fatty acid ester known in the art is contemplated for use in the nanobiocompositions of the present disclosure. Non-limiting examples of fatty acids used include: arachidonic acid, oleic acid, arachidic acid, lauric acid, sad, capric acid, myristic acid, palmitic acid, stearic acid, linoleic acid, linolenic acid, dicaprate, three decanoins, glycerol monoesters, dilaurin, 1-Sunsoft 767, laurocapron (1-dodecyl-aza-cycloheptan-2-one), acylcarnitine, acylcholine, or C1-C 10Alkyl ester (arrcostab) (such as isopropyl myristate IPM), monoglyceride, diglyceride or a pharmaceutically acceptable salt thereof.

[0457] Any suitable synthetic or natural triglyceride known in the art is expected to be used in the nanobiocomposition of the present disclosure. The non-limiting examples of the triglyceride used include: tricaprylin, tristearin, triolein, tripalmitin, 1,2-dipalmitolein, 1,3-dipalmitolein, 1-palmitoyl-3-stearic-2-olein, 1-palmitoyl-2-stearic-3-olein, 2-palmitoyl-1-stearic-3-olein, trilinolein, 1,2-dipalmitoyl-linolein, 1-palmitoyl-dilinolein, 1-stearic-dilinolein, 1,2-diacetyl palmitin, 1,2-distearic-olein, 1,3-distearic-olein, trimyristin, trilaurin and combinations thereof. Suitable triglyceride can be added to the present composition in pure form. Additionally or alternatively, oil and / or process oil containing suitable triglyceride can be added to the composition. Non-limiting examples of oils include coconut oil, corn germ oil, olive oil, palm seed oil, cottonseed oil, palm oil, rapeseed oil, sunflower oil, sperm oil, soybean oil, peanut oil, linseed oil, tall oil, and combinations thereof.

[0458] In the embodiments of the nanobiocompositions provided herein, the nanobiocompositions have a PDI of about 0.01 to about 0.5, including about 0.01, about 0.02, about 0.03, about 0.04, about 0.05, about 0.06, about 0.07, about 0.08, about 0.09, about 0.1, about 0.2, about 0.3, about 0.4 to about 0.5, including all subranges and values ​​therebetween.

[0459] In the embodiments of the nanobio compositions provided herein, the nanobio compositions have a PDI of about 0.1 to about 0.3.

[0460] In embodiments, the diameter of the nanobio composition ranges from about 5 nm to about 400 nm, including about 5 nm, about 6 nm, about 7 nm, about 8 nm, about 9 nm, about 10 nm, about 11 nm, about 12 nm, about 13 nm, about 14 nm, about 15 nm, about 16 nm, about 17 nm, about 18 nm, about 19 nm, about 20 nm, about 21 nm, about 22 nm, about 23 nm, about 24, about 25 nm, about 26 nm, about 27 nm, about 28 nm, about 29, about 30 nm, about 31 nm, about 32 nm, about 33 nm, about 34, about 35 nm, about 36 nm, about 37 nm, about 38 nm, about 39, about 40 nm, about 41 nm, about 42 nm, about 43 nm, about 44 nm, about 45 nm, about 46 nm, about 47 nm, about 48 nm, about 49 nm, about 50 nm, about 51 nm, about 52 nm, about 53 nm, about 54 nm, about 55 nm, about 56 nm, about 57 nm, about 58 nm, about 59 nm, about 60 nm, about 61 nm, about 62 nm, about 63 nm, about 64 nm, about 65 nm about 43 nm, about 44 nm, about 45 nm, about 46 nm, about 47 nm, about 48 nm, about 49 nm, about 50 nm, about 51 nm, about 52 nm, about 53 nm, about 54 nm, about 55 nm, about 56 nm, about 57 nm, about 58 nm, about 59 nm, about 60 nm, about 61 nm, about 62 nm, about 63 nm, about 64 nm, about 65 nm, about 66 nm, about 67 nm, about 68 nm, about 69 nm, about 70 nm, about 71 nm, about 72 nm, about 73 nm, about 74 nm, about 75 nm, about 76 nm, about 77 nm, about 78 nm, about 79 nm, about 80 nm, about 81 nm, about 82 nm, about 83 nm, about 84 nm, about 85 nm, about 86 nm, about 87 nm, about 88 nm, about 89, about 90 nm, about 91 nm, about 92 nm, about 93 nm, about 94, about 95 nm, about 96 nm, about 97 nm, about 98 nm, about 99 nm, about 100 nm, about 101 nm, about 102 nm, about 103 nm, about 104 nm, about 105 nm, about 106 nm, about 107 nm, about 108 nm, about 109 nm, about 110 nm, about 111 nm, about 112 nm, about 113 nm, about 114 nm, about 115 nm, about 116 nm, about 117 nm, about 118 nm, about 119 nm, about 120 nm, about 121 nm, about 122 nm, about 123 nm about 147nm, about 148nm, about 149nm, about 150nm, about 160nm, about 170nm, about 180nm, about 190nm, about 200nm, about 210nm, about 220nm, about 230nm, about 240nm, about 250nm, about 260nm, about 270nm, about 280nm, about 290nm, about 300nm, about 310nm, about 320nm, about 330nm, about 340nm, about 350nm, about 360nm, about 370nm, about 380nm, about 390nm, about 400nm, about 410nm, about 420nm, about 430nm, about 440nm, about 450nm, about 460nm, about 470nm, about 480nm, about 490nm, about 500nm, about 510nm, about 520nm, about 530nm, about 540nmIn some embodiments, the diameter of the nanobiocomposition is about 240 nm, about 250 nm, about 260 nm, about 270 nm, about 280 nm, about 290 nm, about 300 nm, about 310 nm, about 320 nm, about 330 nm, about 340 nm, about 350 nm, about 360 nm, about 370 nm, about 380 nm, about 390 nm, and about 400 nm, including all ranges and values ​​therebetween. In embodiments of the nanobiocompositions provided herein, the diameter of the nanobiocomposition is about 5 nm to about 30 nm, about 5 nm to about 150 nm, about 20 nm to about 150 nm, about 15 nm to about 250 nm, or about 305 nm to about 100 nm. In embodiments of the nanobiocompositions provided herein, the diameter of the nanobiocomposition is about 20 nm to about 100 nm. In embodiments of the nanobiocompositions provided herein, the diameter of the nanobiocomposition is about 25 nm to about 60 nm. In embodiments, the diameter of the nanobiocomposition is measured by dynamic light scattering (DLS).

[0461] In an embodiment, the nanobiocomposition of the present disclosure is disc-shaped.

[0462] In an embodiment, the nanobiocomposition of the present disclosure is spherical.

[0463] In an embodiment, the nanobiocompositions of the present disclosure comprise one or more promoters of trained immunity, or inhibitors of trained immunosuppressants disclosed in US2019 / 0290593, US2020 / 0253884, US2020 / 0376146, WO2018 / 071549, and US2022-0332762, which are incorporated herein by reference in their entirety for all purposes.

[0464] In embodiments, the nanobiocompositions of the present disclosure comprise a trained immune promoter. In embodiments, the trained immune promoter is a Dectin-1 receptor agonist. In embodiments, the trained immune promoter is a NOD2 agonist, such as muramyl dipeptide (MDP), muramyl tripeptide (MTP), or a derivative or prodrug thereof (e.g., mifatide). In embodiments, the trained immune promoter is a derivative of muramyl dipeptide (MDP). In embodiments, the trained immune promoter is muramyl dipeptide phosphatidylethanolamine.

[0465] In embodiments, the nanobiocomposition comprises N-(N-acetylmuramoyl)-L-alanyl-D-α-glutaminyl-N-[(7R)-4-hydroxy-4-oxido-10-oxo-7-[(1-oxohexadecanyl)oxy]-3,5,9-trioxa-4-phosphapentacos-1-yl]-L-alaninamide.

[0466] In embodiments, the nanobiocompositions of the present disclosure comprise a trained immunosuppressant. In embodiments, the trained immunosuppressant is one or more of the trained immunosuppressants disclosed in US2020 / 0376146, which is incorporated herein by reference. In embodiments, the trained immunosuppressant is an mTOR inhibitor. In embodiments, the mTOR inhibitor is rapamycin or a prodrug thereof.

[0467] In an embodiment, the mTOR inhibitor is CCL-779, RAD001 AP23573, C20-methallylrapamycin (C20-Marap), C16-(S)butylsulfonylaminorapamycin (C16-BSrap), C16-(S)-3-methylindorapamycin (C16-iRap) (Bayle et al. Chemistry & Biology 2006, 13:99-107), AZD8055, BEZ235 (NVP-BEZ235), chrysophanic acid chrysophanol), deforolimus (MK-8669), everolimus (RAD0001), KU-0063794, PI-103, PP242, temsirolimus, and WYE-354. In embodiments, the trained immunosuppressant is a rapamycin derivative, for example, a rapamycin derivative disclosed in U.S. Patent Nos. 5,665,772, 6,440,990, 5,985,890, or 6,200,985, each of which is incorporated herein by reference. In embodiments, the compound is 32-deoxyrapamycin, 16-pent-2-ynyloxy-32-deoxyrapamycin, 16-pent-2-ynyloxy-32(S)-dihydro-rapamycin, 16-pent-2-ynyloxy-32(S)dihydro-40-O-(2-hydroxyethyl)-rapamycin, or 40-O-(2-hydroxyethyl)rapamycin. In embodiments, the nanobiocomposition comprises a compound described in WO 94 / 09010, WO 95 / 16691, or WO 96 / 41807, and it has been found that these compounds can be used, for example, as immunosuppressants.

[0468] In embodiments, the sphingolipids of the present disclosure are formulated with any of the nanobiologics disclosed in US2019 / 0290593, US2020 / 0253884, US2020 / 0376146, and WO2018 / 071549, which are incorporated herein by reference in their entirety for all purposes.

[0469] Pharmaceutical formulations containing nanobiocompositions

[0470] When used as a medicament, the nanobiocompositions disclosed herein are typically administered in the form of pharmaceutical compositions. Such compositions can be prepared using methods known in the pharmaceutical art and comprise at least one active compound. In some embodiments, the pharmaceutical composition comprises the nanobiocompositions disclosed herein and a pharmaceutically acceptable carrier. The pharmaceutical compositions disclosed herein may contain more than one type of nanobioagent; for example, a first nanobioagent may have a spherical structure, and a second nanobioagent may have a disc-like structure.

[0471] Typically, the nanobiocompositions of the present disclosure are administered in a pharmaceutically effective amount. The actual amount of compound administered will generally be determined by a physician based on the relevant circumstances, including the condition to be treated, the chosen route of administration, the actual compound administered, the age, weight, and response of the individual patient, and the severity of the patient's symptoms.

[0472] Liquid forms suitable for oral administration may include a suitable aqueous or non-aqueous vehicle with buffers, suspending and dispersing agents, colorants, and flavors, etc. Solid forms may include, for example, any of the following ingredients or compounds of a similar nature: binders such as microcrystalline cellulose, tragacanth, or gelatin; excipients such as starch or lactose; disintegrants such as alginic acid, sodium starch glycolate (Primogel), or corn starch; lubricants such as magnesium stearate; glidants such as colloidal silicon dioxide; sweeteners such as sucrose or saccharin; or flavorings such as mint, methyl salicylate, or orange flavoring.

[0473] Injectable compositions are generally based on injectable sterile saline or phosphate buffered saline or other injectable carriers known in the art. As mentioned above, the active compound in such compositions is usually a minor component, often from about 0.05 to 10% by weight, with the remainder being an injectable carrier or the like.

[0474] The above components for orally administrable, injectable or topically administrable compositions are representative only. Other materials and processing techniques are described in Part 8 of Remington's Pharmaceutical Sciences, 17th ed., 1985, Mack Publishing Company, Easton, Pennsylvania, which is incorporated herein by reference.

[0475] The term "unit dosage form" refers to physically discrete units suitable as unitary dosages for human subjects and other mammals, each unit containing a predetermined quantity of active material calculated to produce the desired therapeutic effect, in association with a suitable pharmaceutical excipient. Typical unit dosage forms include pre-filled, pre-measured ampoules or syringes for liquid compositions or pills, tablets, or capsules in the case of solid compositions.

[0476] Treatment

[0477] Provided herein are methods of treating a subject susceptible to or suffering from immune-related diseases and conditions, including, for example, infectious diseases, sepsis and immunoparalysis in infection, cell proliferative disorders such as cancer, autoimmune and autoinflammatory disorders, transplantation, cardiovascular disease, neurodegenerative diseases, allergies, and other immune-related diseases and conditions.

[0478] Examples of autoimmune diseases include coeliac disease, type I diabetes, multiple sclerosis, thyroiditis, Grave's disease, systemic lupus erythematosus, scleroderma, psoriasis, arthritis, rheumatoid arthritis, alopecia greata, ankylosing spondylitis, Churg-Strauss Syndrome, autoimmune hemolytic anemia, autoimmune hepatitis, Behçet's disease, Crohn's disease, dermatomyositis, glomerulonephritis, Guillain-Barre syndrome, IBD, lupus nephritis, myasthenia gravis, myocarditis, pemphigus / pemphigoid, pernicious anemia, polyarteritis nodosa, polymyositis, primary biliary cirrhosis, rheumatic fever, sarcoidosis, Sjögren's syndrome, ulcerative colitis, uveitis, vitiligo, and Wegener's granulomatosis.

[0479] In embodiments, the nanobiocompositions used herein can be used to treat atherosclerotic lesions, such as coronary atherosclerosis, diabetic atherosclerosis, atherosclerosis, acute coronary syndrome myocardial infarction, angina pectoris, peripheral vascular disease, intermittent claudication.

[0480] In an embodiment, the present disclosure provides a method for inducing transplant tolerance in a subject in need thereof, comprising administering to the subject in need thereof a therapeutically effective amount of a nanobio composition of the present disclosure. In an embodiment, the present disclosure provides a method for preventing organ or tissue rejection in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a nanobio composition of the present disclosure.

[0481] In embodiments, the present disclosure provides methods of treating a subject with a viral, fungal, mycoplasma, bacterial, or protozoal infection comprising administering to the subject a therapeutically effective amount of a nanobiocomposition of the present disclosure.

[0482] In embodiments, the present disclosure provides methods for improving survival of an organ or tissue transplant in a patient in need thereof.

[0483] In embodiments, the transplanted tissue is lung tissue, heart tissue, kidney tissue, liver tissue, retinal tissue, corneal tissue, skin tissue, pancreatic tissue, intestinal tissue, genital tissue, ovarian tissue, bone tissue, tendon tissue, bone marrow, or vascular tissue. In embodiments, the transplanted tissue is a whole organ. In embodiments, the subject is a human, and the organ or tissue transplant is an allogeneic tissue or organ transplant. In embodiments, the nanobiocomposition is administered before, simultaneously with, or after the allogeneic tissue or organ transplant. In certain embodiments, the methods of the present invention further comprise administering to the patient one or more immunosuppressants.

[0484] In an embodiment, the present disclosure provides a method for treating cancer comprising administering to a subject in need thereof a therapeutically effective amount of a nanobio composition of the present disclosure.

[0485] The cancer can be selected from the group consisting of bladder cancer, bone cancer, brain cancer (e.g., glioblastoma multiforme, glioma, astrocytoma), breast cancer, cervical cancer, colon cancer, endometrial cancer, esophageal cancer, eye cancer, head cancer, kidney cancer, liver cancer, lung cancer, mouth cancer, neck cancer, ovarian cancer, pancreatic cancer, prostate cancer (e.g., neuroendocrine prostate cancer), rectal cancer, colorectal cancer, gastric cancer, skin cancer, stomach cancer, testicular cancer, pharyngeal cancer, thyroid cancer, urothelial cancer, and uterine cancer.

[0486] In embodiments, 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, oral 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.

[0487] In embodiments, the cancer is selected from the group consisting of breast cancer, prostate cancer, melanoma, colorectal cancer, lung cancer, pancreatic cancer, and glioblastoma.

[0488] The nanobiocompositions of the present disclosure are contemplated to be administered by a variety of routes including oral, rectal, intraocular, transdermal, subcutaneous, intravenous, intraarterial, intramuscular, intraperitoneal, intradermal, directly into the cerebrospinal fluid, intratracheal, and intranasal. Typically, administration is intravenous.

[0489] In embodiments, training immune promoters or training immune suppressors (e.g., as disclosed herein) can be incorporated into nanobiological preparations or can be physically separated from nanobiological preparations. When physically separated, training immune promoters or training immune suppressors and nanobiological preparations can be administered together, for example, in a suspension, or can be administered separately by the same route of administration or by different routes of administration. When administered separately, training immune promoters or training immune suppressors and nanobiological preparations can be administered within at most 20 minutes, at most 1 hour, at most 2 hours, or within at most 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, or 14 days of each other. In embodiments, training immune promoters and nanobiological preparations can be administered at least 12 hours, at least 1 day, at least 2 days, at least 3 days, at least 7 days, or at least 2 weeks apart.

[0490] The compositions and methods disclosed herein provide two inputs to enhance trained immunity. The nanobiocomposition containing sphingolipids may also contain a trained immune promoter or may be administered together with a trained immune promoter at a sufficiently close time point to obtain a combined effect of the two on the innate immune system. In an embodiment, provided herein is a method for inducing or increasing a trained immune response, the method comprising administering a nanobiocomposition of the present disclosure. In an embodiment, the induction of trained immunity is characterized by enhanced IL-6 production in PBMCs treated with sphingolipid nanobiopreparations after LPS stimulation compared to control PBMCs incubated with culture medium (RPMI) in an in vitro trained immunoassay, for example, as described herein. In an embodiment, the increase in trained immunity is characterized by enhanced IL-6 production in PBMCs treated with sphingolipid nanobiopreparations and HKCA after LPS restimulation compared to control PBMCs treated with HKCA alone in an in vitro trained immunoassay, for example, as described herein.

[0491] In embodiments, provided herein are methods of suppressing a trained immune response comprising administering a nanobio composition of the present disclosure. In embodiments, suppressing or inhibiting trained immunity is characterized by suppressed TNF production in PBMCs treated with a sphingolipid nanobio formulation after LPS restimulation, compared to control PBMCs incubated with culture medium (RPMI) in an in vitro trained immune assay, e.g., as described herein. In embodiments, suppressing or inhibiting trained immunity is characterized by suppressed TNF and / or IL-6 production in PBMCs treated with a sphingolipid nanobio formulation and HKCA after LPS restimulation, compared to control PBMCs treated with HKCA alone in an in vitro trained immune assay.

[0492] In embodiments, the present disclosure provides a method of stimulating or inhibiting a trained immune response in a subject, the method comprising administering (i) a nanobiocomposition comprising:

[0493] (a) apolipoprotein AI (apoA-I) or a peptide mimetic of apoA-I;

[0494] (b) phospholipids; and

[0495] (c) cholesterol;

[0496] (d) Sphingolipids

[0497] wherein sphingolipids are present as about 1-50 mol% of the total lipid composition; and wherein the diameter of the nanobiocomposition is about 8 nm to about 150 nm; and

[0498] (ii) Training of immune promoters or suppressors.

[0499] The nanobiocompositions described herein can be provided in kits. In some embodiments, the kits include (a) the nanobiocompositions described herein, and, optionally, (b) informational materials. The informational materials can be descriptive, instructional, marketing, or other materials related to the methods described herein and / or the uses of the compositions described herein for the methods described herein. In embodiments, the informational materials can include information about the production of the nanobiocompositions. In some embodiments, the informational materials relate to methods of administering the nanobiocompositions. In embodiments, the informational materials can include instructions for administering the nanobiocompositions described herein in a suitable manner to perform the methods described herein, such as in a suitable dose, dosage form, or mode of administration (e.g., a dose, dosage form, or mode of administration described herein). In some embodiments, the informational materials can include instructions for administering the compounds described herein to a suitable subject, such as a human, for example, a human suffering from or at risk for a condition described herein.

[0500] The kit may include one or more containers for a composition containing a nanobio composition described herein. In some embodiments, the kit comprises separate containers, dividers, or compartments for the composition and the informational material. For example, the composition may be contained in a bottle, vial, or syringe, and the informational material may be contained in a plastic sleeve or bag. In some embodiments, the separate elements of the kit are contained within a single, undivided container. For example, the composition is contained in a bottle, vial, or syringe to which the informational material is attached in the form of a label. In some embodiments, the kit comprises a plurality of separate containers (e.g., a pack), each container containing one or more unit dosage forms of a nanobio composition described herein (e.g., a dosage form described herein).

[0501] In embodiments, provided herein are nanobio compositions prepared according to the methods disclosed herein.

[0502] Example

[0503] Example 1: Preparation of sphingolipid-loaded nanobiological preparations

[0504] 1,2-dimyristoyl-sn-glycero-3-phosphocholine (DMPC), cholesterol, and sphingolipids were obtained from Avanti Lipids with a purity of >99%. As previously reported, ApoA-1 was isolated from human HDL concentrate (Biosource Technology) (Braza, MS, et al., Immunity, 2018.49(5): p.819-828e6). DMPC (2.50 mg, 3.69 μmol, 1.0 equivalent), cholesterol (0.29 mg, 0.74 μmol, 0.2 equivalent), and sphingolipids (0.74 μmol, 0.2 equivalent) were placed in a 20 mL vial and dissolved in chloroform (2.0 mL). For the unloaded nanobiological preparation used as a control, sphingolipids were replaced by an additional 0.2 equivalent of DMPC. The solvent was evaporated under vacuum to produce a lipid film, followed by the addition of apoA-1 (1.0 mg, 33 nM) in PBS (5.0 mL). The suspension was sonicated for 7 minutes using a Branson Digital Sonifier SFX150 operating at 60% output power while cooling in an ice-water bath. The slightly opaque solution was concentrated by centrifugal filtration using Vivaspin tubes (Sartorius Biotech, 10 kDa molecular weight cutoff at 4000 rpm and 4°C) until a volume of approximately 1.0 mL remained. PBS (2.0 mL) was added and the sample was centrifuged again until 1.0 mL remained; the process was repeated once more. The resulting solution was filtered through a 0.22 μm polyethersulfone (PES) syringe filter (Celltreat) to obtain the sphingolipid-loaded nanobiological formulation as a white emulsion.

[0505] Characterizing the size and sphingolipid loading of sphingolipid-loaded nanobiologics

[0506] Particle size was determined by dynamic light scattering (DLS) using a Brookhaven Instrument Corporation ZetaPALS analyzer. An aliquot (20 μL) of the nanobiologic was diluted with 1.0 mL PBS and filtered using a 0.22 μm PES syringe filter to remove any dust. Six independent runs of 1 minute each were recorded, and the mean of the number average size distribution was reported. The sphingolipid concentration in the nanobiologic emulsion was determined by1 H NMR measurement. An aliquot (~0.25 mL) of the formulated sphingolipid-loaded nanobiological preparation was freeze-dried and resuspended in a mixture of deuterated chloroform and methanol (50:50 volume%, total 0.5 mL) containing a known concentration of DMPC. The sphingolipid concentration was determined by 1 The samples were analyzed by H NMR and the intensities of signals characteristic of DMPC (CH2-COO) and sphingolipids (double bonds).

[0507] The sphingolipid-loaded nanobiocompositions listed in Table 3 below were prepared. Sizes are reported as the mean of the number-average size distribution. Both nanoparticle size and dispersibility index were determined by dynamic light scattering.

[0508] Table 3: Sphingolipid-loaded nanobiologics.

[0509]

[0510]

[0511]

[0512] Example 2: In vitro evaluation of the effect of exogenously administered sphingolipids on innate immune memory in primary human monocytes

[0513] This study investigated the effect of exogenously administered sphingolipids on innate immune memory in human primary monocytes. To promote efficient delivery of sphingolipids to monocytes, sphingolipids were formulated as in Example 1 and Figure 1C A nanobiologic delivery platform is described that efficiently targets bone marrow cells.

[0514] method

[0515] Human PBMC isolation

[0516] PBMCs were separated by differential centrifugation on Ficoll-Paque (Lymphoprep, StemCell Technologies, Inc.). Cells were washed twice in PBS. PBMCs were resuspended in RPMI culture medium supplemented with 2mM L-alanyl-L-glutamine (glutamax), 1mM pyruvic acid and penicillin / streptomycin (all from Thermo Fisher Scientific) and counted on a Casy counter (Innovatis).

[0517] Lactate dehydrogenase measurement to assess cytotoxicity

[0518] LDH concentrations were measured in the supernatants of PBMCs after 24 h of incubation with small molecule inhibitors or nanoparticles using the CyQuant LDH cytotoxicity assay (Thermo Fisher Scientific). LDH concentrations were calculated as a percentage of the maximum possible LDH concentration in completely lysed cells according to the following formula:

[0519]

[0520] Training experiments with sphingolipid-nanoparticles

[0521] Human PBMCs were isolated and plated as described above. After washing, cells were incubated with medium alone as a negative control or treated with sphingolipid-loaded nanobiological agents at 37°C for 1 hour. 5 Cells / ml HKCA (Invivogen) were incubated with the corresponding sphingolipid-loaded nanoparticles (50 μM) at 37°C for 24 hours. HKCA is a heat-inactivated preparation of Candida albicans. HKCA activates the β-glucan-specific dectin-1 receptor expressed on phagocytes. Here, we use HKCA to promote trained immunity. Subsequently, the cells were washed and allowed to stand for 5 days in RPMI medium containing 10% FBS. After the resting period, the cells were stimulated with RPMI as a negative control, 10 ng / ml LPS (Invivogen) or 1 μg / ml Pam3CSK4 (Invivogen).

[0522] Monocyte isolation

[0523] Use negative MACS to separate and separate monocytes with Pan monocyte separation kit (Miltenyi Biotech).In brief, the PBMC of stimulation is washed with PBS, and hatched 30 minutes with Versene solution (0.48mM EDTA, SigmaAldrich) at 37 ℃.Cells are scraped from flat board, counted, decelerated and resuspended in MACS separation buffer (PBS with 0.5% BSA and 2mM EDTA).Carry out monocyte separation according to the specification sheets of manufacturer.

[0524] Cytokine measurement

[0525] Cytokine production was measured in supernatants using commercial ELISA kits for human TNF, IL-6, IFNγ, and IL-1β (R&D Systems) according to the manufacturer's instructions.

[0526] result

[0527] All tested nanobioformulations were non-toxic to PBMC at the concentrations used ( Figure 3A).

[0528] With sphingolipid-nanobio alone ( Figure 1D 、 Figure 3B and Figure 3C ) or in combination with HKCA ( Figure 1E 、 Figure 3D and Figure 3E ) for 24 h and restimulated with LPS five days later.

[0529] Three sphingolipid-nanobiotins (d18:1 / 16:0 ceramide (Table 3, #4), d18:1 / 24:1 galactosyl(β)ceramide (Table 3, #13), and 24:0 sphingomyelin (Table 3, #16)) enhanced IL-6 production after restimulation, indicating that these nanobiotins induce trained immunity ( Figure 1D 、 Figure 3C Four sphingolipid-nanobioformulations (d18:1 / 16:0 ceramide (Table 3, #4), d18:1 / 24:1 galactosyl(β)ceramidase (Table 3, #13), d18:1 / 24:0 lactosyl(β)ceramide (Table 3, #14), and d18:1 / 18:1 glucosyl(β)ceramide (Table 3, #19)) potentiated HKCA-induced trained immune responses against IL-6 ( Figure 1E 、 Figure 3E ).

[0530] Other sphingolipid-nanobiological agents had the opposite effect. d18:1 / 24:1 lactose (β) ceramide (Table 3, #15) suppressed the TNF response after 5 days of LPS stimulation ( Figure 1D 、 Figure 3C ). Nine sphingolipid-nanobioformulations (20:1 sphingosine (Table 3, #2), d18:1 / 24:0 ceramide (Table 3, #3), d18:0 / 16:0 dihydroceramide (Table 3, #5), d18:1 / 12:0 ceramide-1-phosphate (Table 3, #9), d18:1 / 16:0 and d18:1 / 24:0 galactosyl(β)ceramide (Table 3, #11 and #12), d18:1 / 24:1 lactosyl(β)ceramide (Table 3, #15), 24:0 sphingomyelin (Table 3, #16), and d18:1 / 18:0 glucose(β)ceramide (Table 3, #18)) inhibited HKCA-induced trained immune responses against TNF. 18:1 / 16:0 ceramide-1-phosphate (Table 3, #7) suppressed HKCA training for both TNF and IL-6 responses ( Figure 1E 、 Figure 3D and Figure 3E ).

[0531] These data suggest that exogenously administered sphingolipid-loaded nanobiocompositions can affect innate immune memory. Even short-term exposure to exogenously administered sphingolipid-loaded nanobiocompositions can induce long-term changes in the function of innate immune cells (e.g., about a week, about a month, or longer), indicating a trained immune response. Interestingly, this process depends not only on the type of sphingolipid head group, but also on the saturation state of its fatty acid residues. Species with saturated fatty acid chains (Table 3, #12, #14, #18) have the opposite effect on innate immune memory compared to their cis-monounsaturated counterparts (Table 3, #13, #15, #19). In addition, compared to IL-6 secretion, trained immune suppression by sphingolipid-nanobio preparations has a stronger effect on TNF production. Conversely, the induction of trained immunity was most clearly observed in the IL-6 response.

[0532] Example 3: In vivo evaluation of sphingolipid nanobiocompositions in a B16F10 mouse melanoma model

[0533] In this study, nanobioformulations containing the sphingolipids C24:1 galactosyl (β) ceramide (d18:1 / 24:1 (15Z)) (#13), C16 ceramide (d18:1 / 16:0) (d18:1 / 24:0) (#4), C24:0 lactosyl (β) ceramide (d18:1 / 24:0) (#14), and C18:1 glucosyl (β) ceramide (d18:1 / 18:1 (9Z)) (#19) were tested in the B16F10 melanoma model. In all cases, tumors were inoculated on day 7, and animals were treated with intravenous injections of sphingolipid nanobioformulations at doses of 0.5, 1.5, and 5.0 mg sphingolipids / kg mouse on days 0, 2, and 4 (n=10). Black asterisks indicate the significance of tumor size on that particular day, as determined by one-way ANOVA with Dunnett's multiple comparison analysis. Asterisks in rectangular boxes indicate significance of tumor growth rate. P values ​​were calculated using the Mann-Whitney U test or unpaired t-test. *p<0.05, **p<0.01, ***p<0.001, ns=not significant, as determined by Hather et al., Cancer Inform. 2014, 134, 13974. Results are shown in Figure 2 The data show significant tumor growth reducing properties of sphingolipid nanobiological formulations containing galactosyl(β)ceramide (d18:1 / 24:1(15Z)) (#13) **p<0.01; C24:0 lactosyl(β)ceramide (d18:1 / 24:0) (#14) *p<0.05; and C18:1 glucose(β)ceramide (d18:1 / 18:1(9Z)) **p<0.01 (#19), and at 3 different doses.

[0534] Untreated C57BL / 6 mice were intravenously administered nanobiological formulations at a dose of 0.5 mg / kg for nanobiological formulations #13 and #14 and 5 mg / kg for #19 on days 0, 2, and 4. Two days after the last injection, the animals were sacrificed and blood chemistry analysis was performed. The results are shown in Figure 4 The results were as follows: ALP = alkaline phosphatase, AST = aspartate aminotransferase, ALT = alanine aminotransferase, BUN = blood urea nitrogen, n = 4.

[0535] Preparation #13 89 Zr was radiolabeled (using the procedure reported in van Leent, MMT et al. Sci Adv 7, 1-12, 2021) and administered intravenously to mice bearing B16F10 melanoma. The pharmacokinetics of the radiolabeled nanobiological formulations were then evaluated by ex vivo gamma counting (n=5) for 24 hours ( Figure 5 ) and biodistribution ( Figure 6 ).

[0536] The lipophilic fluorophore DiOC 18 Formulation #13 of was injected intravenously into the B16F10 melanoma mouse model, and the bone marrow of the animals (n=5) was analyzed by flow cytometry 24 hours later ( Figure 7 The results showed that the nanobiologic loaded with Formulation #13 exhibited high uptake in bone marrow cells and hematopoietic organs (n=5). Data are mean ± SD and mean ± SEM for tumor growth experiments.

[0537] Implementation Plan

[0538] 1. A nanobiological composition comprising:

[0539] (a) apolipoprotein AI (apoA-I) or a peptide mimetic of apoA-I;

[0540] (b) sphingolipids; and

[0541] (c) cholesterol;

[0542] wherein the composition is a nanoparticle having a diameter of about 8 nm to about 150 nm.

[0543] 2. The nanobiocomposition of embodiment 1, wherein the sphingolipid is selected from the group consisting of ceramide, sphingomyelin, dihydroceramide, glucosylceramide, sphingosine, sphingosine-1-phosphate, galactosylceramide, ceramide-1-phosphate, lactosylceramide, and mixtures thereof.

[0544] 3. The nanobiocomposition of embodiment 1 or 2, wherein the sphingolipid is selected from the group consisting of:

[0545]

[0546] Sphingosine-1-phosphate (d18:1 / 16:0);

[0547]

[0548] (C16 ceramide (d18:1 / 16:0));

[0549]

[0550] (sphingosine (d20:1));

[0551]

[0552] (C24 ceramide (d18:1 / 24:0));

[0553]

[0554] (C16 dihydroceramide (18:0 / 16:0));

[0555]

[0556] C24 dihydroceramide (d18:0 / 24:0);

[0557]

[0558] C16 ceramide-1-phosphate (d18:1 / 16:0);

[0559]

[0560] C18:1 ceramide-1-phosphate (d18:1 / 18:1(9Z));

[0561]

[0562] C12 ceramide-1-phosphate (d18:1 / 12:0);

[0563]

[0564] C16 galactosyl(α)ceramide (d18:1 / 16:0);

[0565]

[0566] C16 galactosyl(β)ceramide (d18:1 / 16:0);

[0567]

[0568] C24:0 galactosyl(β)ceramide (d18:1 / 24:0);

[0569]

[0570] C24:1 galactosyl(β)ceramide (d18:1 / 24:1);

[0571]

[0572] C24:0 lactose (β) ceramide (d18:1 / 24:0);

[0573]

[0574] C24:1 lactose (β) ceramide (d18:1 / 24:1);

[0575]

[0576] 24:0 sphingomyelin;

[0577]

[0578] 24:1 sphingomyelin;

[0579]

[0580] C18:0 glucosyl(β)ceramide (d18:1 / 18:0); and

[0581]

[0582] C18:1 glucosyl(β)ceramide (d18:1 / 18:1(9Z)).

[0583] 4. The nanobiocomposition of embodiment 3, wherein the sphingolipid is selected from the group consisting of:

[0584]

[0585] (C16 ceramide (d18:1 / 16:0));

[0586]

[0587] (C24:1 galactosyl(β)ceramide (d18:1 / 24:1));

[0588]

[0589] (24:0 sphingomyelin);

[0590]

[0591] (C24:1 galactosyl(β)ceramide (d18:1 / 24:1));

[0592]

[0593] (C24:0 lactose (β) ceramide (d18:1 / 24:0)); and

[0594]

[0595] (C18:1 glucosyl(β)ceramide (d18:1 / 18:1(9Z))).

[0596] The nanobiocomposition according to embodiment 3, wherein the sphingolipid is selected from the group consisting of:

[0597] (sphingosine (d20:1));

[0598]

[0599] (C24 ceramide (d18:1 / 24:0));

[0600]

[0601] (C16 dihydroceramide (18:0 / 16:0));

[0602]

[0603] (C12 ceramide-1-phosphate (d18:1 / 12:0));

[0604]

[0605] (C16 galactosyl(β)ceramide (d18:1 / 16:0));

[0606]

[0607] ((C24:0 galactosyl(β)ceramide (d18:1 / 24:0));

[0608]

[0609] ((C24:1 lactose (β) ceramide (d18:1 / 24:1));

[0610]

[0611] ((24:0 sphingomyelin));

[0612]

[0613] ((C18:0 glucosyl(β)ceramide (d18:1 / 18:0)); and

[0614]

[0615] C16 ceramide-1-phosphate (d18:1 / 16:0).

[0616] 6. The nanobio composition of any one of embodiments 1-5, comprising a phospholipid.

[0617] 7. The nanobiocomposition of embodiment 6, wherein the phospholipid is selected from the group consisting of 1,2-dimyristoyl-sn-glycero-3-phosphocholine (DMPC), 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC), 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC), and mixtures thereof.

[0618] 8. The nanobiocomposition of embodiment 7, wherein the phospholipid is DMPC.

[0619] 9. The nanobiocomposition of any one of embodiments 1-5, comprising a phospholipid and a lysophospholipid.

[0620] 10. The nanobiocomposition of any one of embodiments 1-9, wherein the phospholipid is 1,2-dimyristoyl-sn-glycero-3-phosphocholine (DMPC) or 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC).

[0621] 11. The nanobiocomposition of embodiment 9, wherein the lysophospholipid is selected from the group consisting of 1-myristoyl-2-hydroxy-sn-glycero-3-phosphocholine (MHPC), 1-palmitoyl-2-hydroxy-sn-glycero-3-phosphocholine (PHPC), 1-stearoyl-2-hydroxy-sn-glycero-3-phosphocholine (SHPC), and mixtures thereof.

[0622] 12. The nanobiocomposition of embodiment 9, wherein the phospholipid is 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC) and the lysophospholipid is 1-palmitoyl-2-hydroxy-sn-glycero-3-phosphocholine (PHPC).

[0623] 13. The nanobiocomposition of any one of embodiments 1-12, wherein the nanobiocomposition comprises human apolipoprotein AI (apoA-I).

[0624] 14. The nanobio composition of any one of embodiments 6-13, wherein the sphingolipid is present at about 1-50 mol% of the total lipid composition.

[0625] 15. The nanobio composition of embodiment 14, wherein the sphingolipid is present at about 10-25 mol% of the total lipid composition.

[0626] 16. The nanobio composition of embodiment 15, wherein the sphingolipid is present at about 20 mol% of the total lipid composition.

[0627] 17. The nanobio composition of any one of embodiments 1-16, wherein the cholesterol is present at about 1-30 mol% relative to the phospholipids.

[0628] 18. The nanobio composition of embodiment 17, wherein the cholesterol is present at about 5-25 mol% relative to the phospholipids.

[0629] 19. The nanobio composition of any one of embodiments 1-18, wherein the phospholipids, sphingolipids, and cholesterol are present in a molar ratio of about 1:0.05-0.25:0.05-0.25.

[0630] 20. The nanobiocomposition of any one of embodiments 1-19, wherein the nanobiocomposition has a PDI of about 0.1 to about 0.3.

[0631] 21. The nanobiocomposition of any one of embodiments 1-20, wherein the nanoparticles have a diameter of about 20 nm to about 100 nm.

[0632] 22. The nanobiocomposition of embodiment 21, wherein the nanoparticles have a diameter of about 25 nm to about 60 nm.

[0633] 23. The nanobiocomposition of any one of embodiments 1-22, wherein the nanoparticles are spherical.

[0634] 24. The nanobiocomposition of any one of embodiments 1-22, wherein the nanoparticles are disc-shaped.

[0635] 25. The nanobiocomposition according to any one of embodiments 1-24, wherein the nanobiocomposition is suitable for intravenous or intraarterial administration.

[0636] 26. A method for treating cancer in a subject in need thereof, comprising administering to the subject an effective amount of the nanobio composition of any one of embodiments 1-25.

[0637] 27. The method of embodiment 26, 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, throat cancer, thyroid cancer, urothelial cancer, and uterine cancer.

[0638] 28. The method of embodiment 27, wherein the cancer is selected from the group consisting of breast cancer, prostate cancer, melanoma, colorectal cancer, lung cancer, pancreatic cancer, and glioblastoma.

[0639] 29. The method of any one of embodiments 26-28, wherein the nanobiocomposition is administered intravenously or intraarterially.

[0640] 30. The method of any one of embodiments 26-29, wherein the subject is human.

[0641] 31. A method of stimulating a trained immune response in a subject comprising administering

[0642] (i) the nanobiocomposition of any one of embodiments 1-25; and

[0643] (ii) Training immune boosters.

[0644] 32. The method of embodiment 31, wherein the trained immune promoter is a Dectin-1 receptor agonist, a NOD-2 agonist, or a combination thereof.

[0645] 33. The method of embodiment 31, wherein the nanobiocomposition comprises a trained immune enhancer.

[0646] 34. The method of embodiment 31, wherein the nanobiocomposition and the trained immune enhancer are administered in separate compositions.

[0647] 35. The method of embodiment 34, wherein the nanobiocomposition and the training immune enhancer are administered within at most 1, 2, 3, 4, 5, 6, or 7 days of each other.

[0648] 36. The method of embodiment 34 or 35, wherein the routes of administration are the same.

[0649] 37. The method of embodiment 36, wherein the route is intravenous.

[0650] 38. The method of embodiment 34 or 35, wherein the routes of administration are different.

[0651] 39. The method of embodiment 38, wherein the nanobiocomposition is administered intravenously.

[0652] 40. The method of embodiment 31, wherein the trained immune enhancer is a Dectin-1 receptor agonist.

[0653] 41. The method of embodiment 40, wherein the Dectin-1 receptor agonist is HKCA (heat-killed Candida albicans).

[0654] 42. The method of embodiment 31, wherein the trained immune enhancer is a NOD-2 agonist.

[0655] 43. The method of embodiment 42, wherein the NOD2-agonist is muramyl dipeptide (MDP), muramyl tripeptide (MTP), or a derivative or prodrug thereof.

Claims

1. A nanobiological composition comprising: (a) apolipoprotein AI (apoA-I) or a peptide mimetic of apoA-I; (b) sphingolipids; and (c) cholesterol; The composition is a nanoparticle having a diameter of about 8 nm to about 150 nm.

2. The nanobiocomposition according to claim 1, wherein the sphingolipid is selected from the group consisting of ceramide, sphingomyelin, dihydroceramide, glucosylceramide, sphingosine, sphingosine-1-phosphate, galactosylceramide, ceramide-1-phosphate, lactosylceramide and mixtures thereof.

3. The nanobio composition of claim 2, wherein the sphingolipid is selected from the group consisting of glucosylceramide, galactosylceramide, or lactosylceramide.

4. The nanobiocomposition according to claim 3, wherein the sphingolipid is glucosylceramide or galactosylceramide.

5. The nanobiocomposition of claim 4, wherein the sphingolipid is glucosylceramide.

6. The nanobio composition of claim 4, wherein the sphingolipid is galactosylceramide.

7. The nanobiocomposition according to claim 1, wherein the sphingolipid is of formula (I): or a pharmaceutically acceptable salt thereof, wherein: R 1 is a saturated aliphatic chain or an unsaturated aliphatic chain; R 2 It is H, R 4 , or C(O)R 4 ; R 3 It is H, sugar, P(O)(OH)2, or P(O)(OH)O(CH2) n R 5 , where n = 1-10; R 4 is a saturated aliphatic chain or an unsaturated aliphatic chain; and R 5 It is H, NH2, N(CH3)3 + , OH, or sugar.

8. The nanobiocomposition according to claim 7, wherein R 1 It is unsaturated C 14-24 Aliphatic chain, R 2 It is C(O)R 4 ; R 3 yes and R 4 It is C 11-30 Unsaturated fatty chain or C 11-30 Saturated aliphatic chain.

9. The nanobiocomposition according to claim 8, wherein R 1 It is unsaturated C 15-17 Aliphatic chain.

10. The nanobiocomposition according to claim 9, wherein R 1 It is unsaturated C 15-17 Aliphatic chain; R 2 It is C(O)R 4 ; R 3 yes and R 4 It is C 11-30 Unsaturated fatty chain.

11. The nanobiocomposition according to any one of claims 7 to 10, wherein R 1 It is C 15 Alkenyl.

12. The nanobiocomposition according to any one of claims 7 to 11, wherein R 4 It is C 14-24 Unsaturated fatty chain or C 14-24 Saturated aliphatic chain.

13. The nanobiocomposition according to claim 12, wherein R 4 It is C 17 Alkenyl or C 23 Alkenyl.

14. The nanobiocomposition according to any one of claims 7 to 13, wherein R 3 yes 15. The nanobio composition of claim 1 or 2, wherein the sphingolipid is selected from the group consisting of:

16. The nanobio composition of claim 15, wherein the sphingolipid is selected from the group consisting of:

17. The nanobio composition of claim 15, wherein the sphingolipid is selected from the group consisting of:

18. The nanobio composition according to any one of claims 1 to 17, comprising a phospholipid.

19. The nanobio composition of claim 18, wherein the phospholipid is selected from the group consisting of 1,2-dimyristoyl-sn-glycero-3-phosphocholine (DMPC), 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC), 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC), and mixtures thereof.

20. The nanobio composition of claim 19, wherein the phospholipid is DMPC.

21. The nanobio composition according to any one of claims 1-17, comprising phospholipids and lysophospholipids.

22. The nanobio composition of any one of claims 1-21, wherein the phospholipid is 1,2-dimyristoyl-sn-glycero-3-phosphocholine (DMPC) or 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC).

23. The nanobio composition of claim 21, wherein the lysophospholipid is selected from the group consisting of 1-myristoyl-2-hydroxy-sn-glycero-3-phosphocholine (MHPC), 1-palmitoyl-2-hydroxy-sn-glycero-3-phosphocholine (PHPC), 1-stearoyl-2-hydroxy-sn-glycero-3-phosphocholine (SHPC), and mixtures thereof.

24. The nanobio composition of claim 21, wherein the phospholipid is 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC) and the lysophospholipid is 1-palmitoyl-2-hydroxy-sn-glycero-3-phosphocholine (PHPC).

25. The nanobiocomposition of any one of claims 1-24, wherein the nanobiocomposition comprises human apolipoprotein AI (apoA-I).

26. The nanobio composition of any one of claims 18-25, wherein the sphingolipid is present at about 1-50 mol% of the total lipid composition.

27. The nanobio composition of claim 26, wherein the sphingolipid is present at about 10-25 mol% of the total lipid composition.

28. The nanobio composition of claim 27, wherein the sphingolipid is present at about 20 mol% of the total lipid composition.

29. The nanobio composition of any one of claims 1-28, wherein the cholesterol is present at about 1-30 mol% relative to the phospholipids.

30. The nanobio composition of claim 29, wherein the cholesterol is present at about 5-25 mol% relative to the phospholipids.

31. The nanobio composition of any one of claims 1-30, wherein the phospholipids, sphingolipids, and cholesterol are present in a molar ratio of about 1:0.05-0.25:0.05-0.

25.

32. The nanobio composition of any one of claims 1-31, wherein the nanobio composition has a PDI of about 0.1 to about 0.

3.

33. The nanobio composition of any one of claims 1-32, wherein the nanoparticles have a diameter of about 20 nm to about 100 nm.

34. The nanobio composition of claim 33, wherein the nanoparticles have a diameter of about 25 nm to about 60 nm.

35. The nanobio composition of any one of claims 1-34, wherein the nanoparticles are spherical.

36. The nanobio composition of any one of claims 1-35, wherein the nanoparticles are disc-shaped.

37. The nanobio composition of any one of claims 1-36, wherein the nanobio composition is suitable for intravenous or intraarterial administration.

38. A method for treating cancer in a subject in need thereof, comprising administering to the subject an effective amount of the nanobio composition of any one of claims 1-37.

39. The method of claim 38, 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, cervical cancer, ovarian cancer, pancreatic cancer, prostate cancer, rectal cancer, colorectal cancer, skin cancer, stomach cancer, testicular cancer, pharyngeal cancer, thyroid cancer, urothelial cancer, and uterine cancer.

40. The method of claim 39, wherein the cancer is selected from the group consisting of breast cancer, prostate cancer, melanoma, colorectal cancer, lung cancer, pancreatic cancer, and glioblastoma.

41. The method of any one of claims 38-40, wherein the nanobiocomposition is administered intravenously or intraarterially.

42. The method of any one of claims 38-41, wherein the subject is a human.

43. A method of stimulating a trained immune response in a subject comprising administering (i) a nanobio composition according to any one of claims 1 to 37; and (ii) Training immune boosters.

44. The method of claim 43, wherein the trained immune promoter is a Dectin-1 receptor agonist, a NOD-2 agonist, or a combination thereof.

45. The method of claim 43, wherein the nanobiocomposition comprises the trained immune enhancer.

46. ​​The method of claim 43, wherein the nanobio composition and the trained immune enhancer are administered in separate compositions.

47. The method of claim 46, wherein the nanobiocomposition and the trained immune enhancer are administered within at most 1, 2, 3, 4, 5, 6, or 7 days of each other.

48. The method of claim 46 or 47, wherein the routes of administration are the same.

49. The method of claim 48, wherein the route is intravenous.

50. The method of claim 46 or 47, wherein the routes of administration are different.

51. The method of claim 50, wherein the nanobiocomposition is administered intravenously.

52. The method of claim 43, wherein the trained immune enhancer is a Dectin-1 receptor agonist.

53. The method of claim 52, wherein the Dectin-1 receptor agonist is HKCA (heat-killed Candida albicans).

54. The method of claim 43, wherein the trained immune enhancer is a NOD-2 agonist.

55. The method of claim 54, wherein the NOD2-agonist is muramyl dipeptide (MDP), muramyl tripeptide (MTP), or a derivative or prodrug thereof.

56. The method of claim 54, wherein the NOD2-agonist is the muramyl tripeptide phosphatidylethanolamine.

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