Lipoprotein-like nanoparticles with an organic core and use thereof

Abstract

The present disclosure relates, in general, to synthetic, spherical lipoprotein-like nanoparticles comprising an organic core material that provides properties that are similar to mature human HDLs and use thereof to treat disorders such as cancer or immune disorders. In various embodiments, the disclosure provides a spherical lipoprotein-like nanoparticle (HDL-NP) comprising: a core and a shell surrounding and attached to the core, wherein the core comprises a lipid conjugated organic scaffold comprising a dendritic lipid scaffold.

Description

LIPOPROTEIN-LIKE NANOPARTICLES WITH AN ORGANIC CORE AND USE THEREOFCROSS REFERENCE TO RELATED APPLICATIONS

[0001] The present application claims the priority benefit of U.S. Provisional Patent Application No. 63 / 534,038, filed August 22, 2023, herein incorporated by reference in its entirety.FIELD

[0002] The present disclosure relates, in general, to lipoprotein-like nanoparticles (HDL- NP) comprising an organic core material that provides properties that are consistent with mature human HDLs and use thereof to treat disorders such as cancer or immune disorders.BACKGROUND

[0003] Nanoparticles are in the submicron size domain and possess unique sizedependent properties that make the materials superior compared to their bulk forms. The advanced chemical and physical properties associated with nanoparticles have led to their extensive use in the fields of biology and medicine. High-density lipoproteins (HDL) are dynamic nanoparticles that circulate in the blood.

[0004] HDLs have been associated with atherosclerotic cardiovascular disease (ASCVD) (Mendivil et al., Arterioscler Thromb Vase Biol 2016, 36 (1 ), 156-65). Synthetic HDLs (e.g., CER-001 ) have been investigated as therapeutics to treat cardiovascular disease with mixed success (Andrews et al., Cardiovasc Diagn Ther 2017, 7 (1), 45-51 ; Kataoka, et al., Cardiovasc Diagn Ther 2017, 7 (3), 252-263; Keyserling et al., Clin Drug Investig 2017, 37 (5), 483-491).SUMMARY

[0005] The present disclosure provides synthetic lipoprotein-like nanoparticles (HDL NPs) comprising an organic scaffold linked to a phospholipid tail and methods for synthesis and characterization thereof. The particles are synthesized using lipid-conjugated organic scaffolds, including pentaerythritol distearate (PE-S2), pentaerythritol dioleate (PE-O2), pentaerythritol tetrastearate (PE-S4), pentaerythritol tetraoleate (PE-O4), sorbitol(polyethylene glycol)n-S6 (Sorb(PEG)n-S6), sorbitol(polyethylene glycol)n-06 (Sorb(PEG)n-06), tetrastearate (S4), and tetraoleate (04) cores.

[0006] In various embodiments, the disclosure provides a spherical lipoprotein-like nanoparticle (HDL-NP) comprising: a core and a shell surrounding and attached to the core, wherein the core comprises a lipid conjugated organic scaffold comprising a dendritic lipid scaffold. The dendritic lipid scaffold comprises tunable hydrophilic, hydrophobic oramphiphilic molecule cores that can be used to facilitate 3-D supramolecular assembly of other core and / or shell lipids to form lipid nano- and micro-particles.

[0007] In various embodiments, the dendritic lipid scaffold comprises pentaerythritol distearate (PE-S2), pentaerythritol dioleate (PE-O2), pentaerythritol tetrastearate (PE-S4), pentaerythritol tetraoleate (PE-O4), sorbitol(polyethylene glycol)n-hexasterate (Sorb(PEG)n- Se) sorbitol(polyethylene glycol)n-hexaoleate (Sorb(PEG)n-Oe), tetraoleate (O4) or tetrastearate (S4).

[0008] In various embodiments, the shell is a lipid shell. In some embodiments, the shell is a lipid bilayer or monolayer.

[0009] In various embodiments, the spherical HDL-NP has a surface zeta potential of about -5 to about -60 millivolts (mV).

[0010] In various embodiments, the organic core comprises a headgroup-modified fatty acid. In some embodiments, the headgroup-modified fatty acid comprises C12. C14, C16, C18 or C20 tails. In some embodiments, the phospholipid is selected from the group consisting of 1 ,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC), 1 ,2-dioleoyl-sn-glycero-3- phosphocholine (DOPC), 1 ,2-diolinoleoyl-sn-glycero-3-phosphocholine (DLPC), 1 ,2-dioleoyl- sn-glycero-3-phospho-L-serine (DOPS), 1 ,2-distearoyl-sn-glycero-3-phospho-(1 '-rac- glycerol) (18:0 PG), or 1 ,2-dilinoleoyl-sn-glycero-3-phospho-(1 '-rac-glycerol) (18:2 PG).

[0011] In various embodiments, the components of the shell are selected from the group consisting of ionizable derivatives of lauric, myristic, palmitic, stearic, and arachidic acids and / or their unsaturated derivatives (oleic, linoleic, linolenic, eleostearic, and arachidonic acids), which can be mono- or polyunsaturated. Such derivatives can be salts of the fatty acids or the acid themselves being ionized in situ and assembled under the appropriate conditions. In various embodiments, the shell components comprise 1 ,2-dipalmitoyl-sn- glycero-3-phosphocholine (DPPC), 1 ,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC), 1 ,2- diolinoleoyl-sn-glycero-3-phosphocholine (DLPC), 1 ,2-dioleoyl-sn-glycero-3-phospho-L- serine (DOPS), 1 ,2-distearoyl-sn-glycero-3-phospho-(1 '-rac-glycerol) (18:0 PG), or 1 ,2- dilinoleoyl-sn-glycero-3-phospho-(1'-rac-glycerol) (18:2 PG).

[0012] In various embodiments, the organic core HDL nanoparticles (oc-HDL NP) provided herein include DPPC Polyethylene glycol) sorbitol hexaoleate (Sorb(PEG)l -Oe), DPPC Polyethylene glycol) sorbitol hexastearate (Sorb(PEG)l-Se), DPPC PE-S4, DPPC PE-O4, DOPC PE-O4 and DOPS PE-O4.

[0013] In various embodiments, a small molecule with a plurality of terminal functional groups is coupled to one or more lipids to form the dendritic lipid scaffold. In variousembodiments, the small molecule is pentaerythritol, sorbitol and tetraA7s(4-functionalized phenyl)methane. In various embodiments, the plurality of functional groups are hydroxyls. In various embodiments, the plurality of functional groups are amino groups. In various embodiments, the plurality of functional groups are carboxylates. In some embodiments, the fatty acids can be reduced to the corresponding alcohols, e.g., lauryl, stearyl, and oleyl alcohols, and coupled to the small molecule core to form the dendritic lipid scaffold.

[0014] In various embodiments, the spherical HDL-NP has a diameter of about 5-60 nm, about 10-50 nm, about 5-40 nm, about 10-30 nm, about 5-30 nm, about 5-20 nm, about 5-15 nm, about 5-10 nm, about 8-13 nm, or about 8-12 nm. In various embodiments, the spherical HDL-NP has a negative zeta potential similar to or more negative than human HDL.

[0015] In various embodiments, the zeta potential is about -5 mV to about -25 mV. In some embodiments, the zeta potential is from about -5 mV to about -30 mV, about -10 mV to about -20 mV or about -5 mV to about -15 mV. In some embodiments, the zeta potential is about -5 mV, about -6 mV, about -7 mV, about -8 mV, about -9 mV, about -10 mV, about -11 mV, about -12 mV, about -13 mV, about -14 mV, about -15 mV, about -16 mV, about -17 mV, about -18 mV, about -19 mV, about -20 mV, about -21 mV, about -22 mV, about -23 mV, about -24 mV, about -25 mV, about -26 mV, about -27 mV, about -28 mV, about -29 mV or about -30 mV.

[0016] In various embodiments, the HDL-NP is capable of associating with cholesterol. In various embodiments, the spherical HDL-NP has a more efficient cholesterol transport capacity than that of apolipoprotein or a synthetic HDL nanoparticle with a gold core.

[0017] In various embodiments, the spherical HDL-NP further comprises apolipoprotein. In some embodiments, the apolipoprotein is apolipoprotein A, apolipoprotein B, apolipoprotein C, apolipoproteins D, apolipoproteins E or apolipoproteins H. In various embodiments, the apolipoprotein is selected from the group consisting of apo A-l, apo A-l I, apo A-IV, apo A-V, apo B48, apo B100, apo C-l, apo C-ll, apo C-lll, apo C-IV, apolipoprotein D, apolipoprotein E and apolipoprotein H. In various embodiments, the apolipoprotein is apolipoprotein A-l (ApoA1 ).

[0018] In various embodiments, the HDL-NP has a hydrodynamic diameter of greater than 8 nm, e.g., from about 8 nm to about 30 nm. In various embodiments, the HDL-NP has a hydrodynamic diameter of 8 nm-17 nm. In various embodiments, the HDL-NP has a hydrodynamic diameter of 10 nm-15 nm. In various embodiments, the HDL-NP has a hydrodynamic diameter of 12 nm-14nm.

[0019] In various embodiments, the HDL-NP organic core comprises PE-S4 and has a diameter of about 10 nm to about 20nm and a zeta potential between about -10 mV to about -20 mV.

[0020] In various embodiments, the HDL-NP organic core comprises PE-O4 and has a diameter of about 8 nm to about 17 nm and a zeta potential between about -5 mV to about - 15 mV.

[0021] In various embodiments, the HDL-NP organic core comprises Sorb(PEG)n-Se and has a diameter of about 10 nm to about 30 nm and a zeta potential between about -10 mV to about -20 mV.

[0022] In various embodiments, the HDL-NP organic core comprises Sorb(PEG)n-Oe and has a diameter of about 10 nm to about 25 nm and a zeta potential between about -15 mV to about -25 mV.

[0023] In various embodiments, the HDL-NP organic core comprises S4 and has a diameter of about 7 nm to about 25 nm and a zeta potential between about -10 mV to about -20 mV.

[0024] In various embodiments, the HDL-NP organic core comprises O4 and has a diameter of about 7 nm to about 25 nm and a zeta potential between about -10 mV to about -20 mV.

[0025] In various embodiments, the spherical HDL-NP further comprises a therapeutic agent or natural lipophilic molecule adsorbed or linked to the HDL-NP. In some embodiments, the natural lipophilic molecule is vitamin D, vitamin A, vitamin K or vitamin E.

[0026] In various embodiments, the therapeutic agent is a therapeutic nucleic acid, an anti-cancer agent, a chemotherapeutic agent, an antioxidant or an anti-inflammatory agent. Therapeutic agents contemplated herein are described more fully in the Detailed Description.

[0027] Also contemplated herein is a pharmaceutical composition comprising a spherical HDL-NP as described herein and a pharmaceutically acceptable excipient, diluent or carrier.

[0028] The disclosure also contemplates a method for treating a cancer, the method comprising: administering to a subject having a cancer an effective amount of a spherical lipoprotein-like nanoparticle (HDL-NP) comprising: a core and a shell surrounding and attached to the core, wherein the core comprises a lipid conjugated organic scaffold comprising a dendritic lipid scaffold.

[0029] Also provided is a method of treating an inflammatory disorder comprising, administering to a subject having an inflammatory disorder an effective amount of a spherical lipoprotein-like nanoparticle (HDL-NP) comprising: a core and a shell surrounding and attached to the core, wherein the core comprises a lipid conjugated organic scaffold comprising a dendritic lipid scaffold.

[0030] In various embodiments, the lipid conjugated organic scaffold comprises pentaerythritol distearate (PE-S2), pentaerythritol dioleate (PE-O2), pentaerythritol tetrastearate (PE-S4), pentaerythritol tetraoleate (PE-O4), sorbitol(polyethylene glycol)n-S6 (Sorb(PEG)n-Se) or sorbitol(polyethylene glycol)n-06 (Sorb(PEG)n-Oe), tetrastearate (S4) or tetraoleate (O4).

[0031] In various embodiments, provided is a pharmaceutical composition comprising a spherical HDL-NP as described herein.

[0032] In various embodiments, the subject is a mammal. In various embodiments, the subject is a human.

[0033] In various embodiments, the spherical HDL-NP is administered intravenously, subcutaneously, intramuscularly, or orally.

[0034] The disclosure further contemplates a method of reducing NF-kB and / or TNF-a activity, comprising administering to a subject an effective amount of a composition comprising a spherical HDL-NP as described herein. In various embodiments, the reduction NF-kB and / or TNF-a activity is in a subject having cancer, an inflammatory disease, a cardiovascular disease, a bone disease, or an immune disease.

[0035] In another aspect, the disclosure contemplates a method for making a spherical lipoprotein-like nanoparticle (HDL-NP) comprising:

[0036] -preparing an organic scaffold solution;

[0037] -contacting the organic scaffold solution with apolipoprotein to generate an organic scaffold-Apo solution;

[0038] -contacting the organic-Apo solution with a lipid solution to generate an organic- Apo-lipid solution;

[0039] -admixing the organic -Apo-lipid solution by vortexing and / or sonication to form HDL-NP;

[0040] -isolating the HDL-NP.

[0041] In various embodiments, the method generates an HDL-NP comprising a core and a shell surrounding and attached to the core, wherein the core comprises a lipid conjugated organic scaffold.

[0042] In various embodiments, the organic scaffold solution comprises any one of pentaerythritol distearate (PE-S2), pentaerythritol dioleate (PE-O2), pentaerythritol tetrastearate (PE-S4), pentaerythritol tetraoleate (PE-O4), sorbitol(polyethylene glycol)n-S6 (Sorb(PEG)n-Se) or sorbitol(polyethylene glycol)n-06 (Sorb(PEG)n-Oe) tetrastearate (S4) or tetraoleate (O4).

[0043] In various embodiments of the method, the organic core comprises a headgroup- modified fatty acid. In various embodiments, the headgroup-modified fatty acid comprises C12, C14, C16, C18 or C20 tails. In various embodiments, the fatty acid is monounsaturated or polyunsaturated.

[0044] In various embodiments, the lipid components of the organic scaffold are selected from the group consisting of lauric acid, myristic acid, palmitic acid, stearic acid, arachidic acid, oleic acid, linoleic acid, linolenic acid, eleostearic acid, and arachidonic acid.

[0045] In various embodiments, the apolipoprotein is apolipoprotein A-l.

[0046] In various embodiments, the shell comprises components selected from the group consisting of 1 ,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC), 1 ,2-dioleoyl-sn-glycero-3- phosphocholine (DOPC), 1 ,2-diolinoleoyl-sn-glycero-3-phosphocholine (DLPC), 1 ,2-dioleoyl- sn-glycero-3-phospho-L-serine (DOPS), 1 ,2-distearoyl-sn-glycero-3-phospho-(1'-rac- glycerol) (18:0 PG), or 1 ,2-dilinoleoyl-sn-glycero-3-phospho-(1 '-rac-glycerol) (18:2 PG).

[0047] In various embodiments, the organic core HDL nanoparticles (oc-HDL NP) provided herein include DPPC Polyethylene glycol) sorbitol hexaoleate (Sorb(PEG)l -Oe), DPPC Polyethylene glycol) sorbitol hexastearate (Sorb(PEG)l-Se), DPPC PE-S4, DPPC PE-O4, DOPC PE-O4 and DOPS PE-O4.

[0048] It is understood that each feature or embodiment, or combination, described herein is a non-limiting, illustrative example of any of the aspects of the invention and, as such, is meant to be combinable with any other feature or embodiment, or combination, described herein. For example, where features are described with language such as “one embodiment”, “some embodiments”, “certain embodiments”, “further embodiment”, “specific exemplary embodiments”, and / or “another embodiment”, each of these types of embodiments is a non-limiting example of a feature that is intended to be combined with any other feature, or combination of features, described herein without having to list every possible combination. Such features or combinations of features apply to any of the aspectsof the invention. Where examples of values falling within ranges are disclosed, any of these examples are contemplated as possible endpoints of a range, any and all numeric values between such endpoints are contemplated, and any and all combinations of upper and lower endpoints are envisioned.BRIEF DESCRIPTION OF THE DRAWINGS

[0049] Figure 1 shows a schematic of synthesis of the Sorb(PEG)i-Se organic scaffold.

[0050] Figure 2 shows a schematic of synthesis of the tetrakis(4-oleyl benzoate) methane core scaffold.

[0051] Figure 3 shows a schematic of synthesis of the tetrakis(4-stearyl benzoate) methane core (S4 core) scaffold.

[0052] Figure 4 shows a schematic of synthesis of the tetraoleyl pentaerythritol core (Pe- 04 core) scaffold.

[0053] Figure 5 shows an exemplary synthesis schematic of an oc-HDL-NP and characterization of oc-HDL NPs, demonstrating all cores form small (12-20 nm), negatively charged, spherical structures that can bind apolipoprotein.

[0054] Figure 6 shows that oc-HDL NPs transport cholesterol from J774 macrophages to HepG2 hepatocytes.

[0055] Figure 7 shows oc-HDL NPs load cholesteryl esters.

[0056] Figure 8 demonstrates oc-HDL NPs reduce inflammation in human macrophage cells.

[0057] Figure 9 shows representative transmission electron microscopy (TEM) images of oc-HDL NPs. Scale bar = 50 nm.

[0058] Figures 10A-10B show cholesterol efflux by oc-HDL NP. Percent of cholesterol efflux was determined after 6-hour incubation of oc-HDL NPs with J774 macrophages either in the presence or absence of cAMP (Fig. 10A). (one-way ANOVA, p < 0.05). Western blot analysis for ABCA1 after 24 hours of cholesterol efflux by oc-HDL NPs. p-actin was used as a loading control (Fig. 10B).

[0059] Figure 11 shows oc-HDL NPs suppress pro-inflammatory molecules NF-KB (right) and JNK (left) signaling in THP-1 cells. One-way ANOVA (p < 0.05).

[0060] Figure 12 shows oc-HDL NPs support cholesteryl ester loading. One-way ANOVA (p < 0.05).DETAILED DESCRIPTION

[0061] The synthesis involved the supramolecular assembly of the present HDL NPs using lipid-conjugated organic scaffolds, apolipoprotein-A1 (Apo-A1), and phospholipids, provide the opportunity to tune particle size, shape, composition, and charge. The biological activities of the lipid-conjugated oc-HDL NPs were measured by assessing HDL function using in vitro models of reverse cholesterol transport (RCT), cholesteryl ester loading, and immunomodulation. Data demonstrate that all oc-HDL NPs possess RCT potential, cholesterol loading capability, and anti-inflammatory properties. As existing HDL-based therapeutics have faced limited translational success, in part due to the lack of access to HDLs with well-defined structural and chemical properties, it is envisioned that these biomimetic nanoparticles will provide the opportunity to better understand the structural- functional relationships of HDL and therapeutics for the prevention and treatment of cardiovascular disease, inflammatory diseases, and other chronic diseases.Lipoprotein-like Nanoparticles

[0062] Native HDLs are circulating nanoparticles (-8-13 nm in diameter) that transport cholesterol and play important roles in cancer and cardiovascular disease. Spherical HDLs are the most abundant HDL sub-species, and are particularly important clinically. However, previous HDL mimics have failed to reproduce features of spherical HDL (e.g., spherical HDL conformation) without relying on inorganic template materials (e.g., gold nanoparticles). Herein, it was unexpectedly found that the spherical HDL conformation could be achieved in HDL-like nanoparticles having lipid-conjugated organic scaffolds. These HDL-like nanoparticles of the present disclosure are the first of their kind, namely spherical HDL mimics with a soft material core.

[0063] The HDL-like nanoparticles of the present disclosure mimic spherical HDL species using tunable, lipid-conjugated organic scaffolds. The core design motif constrains and orients phospholipid geometry to facilitate the assembly of soft-core nanoparticles that are, resemble human HDLs in their size, shape, surface chemistry, composition, and protein secondary structure. The HDL-like nanoparticles mimic the structure of native HDL with respect to size (~10 nm), surface chemistry (-20 mV zeta potential), and HDL protein secondary structure as determined by circular dichroism. Synthetic HDLs have demonstrated promise as therapy for cardiovascular disease and cancer, among other indications. Two of the primary features of synthetic HDL mimics which are required for recapitulating salient HDL functions are 1) a soft material core, and 2) a spherical conformation. The HDL-like nanoparticles of the present disclosure are designed to enable superior therapeutic efficacy when compared to previous synthetic platforms, none of which reproduce both of the features noted above.

[0064] Clinically, elevated HDL cholesterol levels are associated with reduced risk of atherosclerotic cardiovascular disease (ASCVD) (Roth, et al. Circulation 2015, 132 (17), 1667-78; Shepard, et al. Circ Cardiovasc Qual Outcomes 2015, 8 (4), 455-6.)). At the cellular level, HDLs target macrophages and hepatocytes to remove and deliver cholesterol, respectively, which has been shown to reduce inflammation and atherosclerotic burden (Feig, et al. PNAS USA 2011 , 108 (17), 7166-71 ; Yvan-Charvet, et al. Arterioscler Thromb Vase Biol 2010, 30 (2), 139-43.). Due to the beneficial association between HDL and ASCVD, cell-specific targeting properties, and the intrigue of using HDLs for targeted drug delivery, tremendous effort has focused on synthesizing HDLs that resemble their natural counterparts (Lacko et al. Front Pharmacol 2015, 6, 247).

[0065] There are multiple HDL species in the blood that serve as synthetic targets. Immature HDLs are cholesterol poor and discoidal in shape, consisting mainly of the HDL- defining protein, apolipoprotein A-l (which may be referred to herein as apoA-l, apoA-1 , and / or apolipoprotein A-1 ), and phospholipids. Mature forms of HDL, which also harbor apoA-l and a surface layer of phospholipids, are cholesterol and cholesteryl ester rich, and assume more spherical shapes. Spherical HDLs comprise the majority of HDLs in circulation (Rosenson, et al. Clin Chem 2011 , 57 (3), 392-410; Kontushet al. Pharmacol Rev 2006, 58 (3), 342-74.). Yet, synthetic forms of HDL used as therapy have exclusively resembled immature discoidal HDL. This is largely because these recombinant forms of HDL (rHDL) are relatively easy to make by self-assembling phospholipids and apoA-l (Simonsen, J. B. Nanomedicine 2016, 12 (7), 2161-2179.; Cao, et al. Drug Discov Today 2017, 22 (1), ISO- 185; Andrews et al. Cardiovasc Diagn Ther 2017, 7 (1), 45-51 ; Kataokaet al., Cardiovasc Diagn Ther 2017, 7 (3), 252-263; Keyserling, et al. Clin Drug Investig 2017, 37 (5), 483-491). Clinical trials using rHDLs have shown some promise but reveal that there is significant room for improvement with regard to making more potent and effective synthetic HDLs (Andrews et al. Cardiovasc Diagn Ther 2017, 7 (1), 45-51 ; Kataokaet al., Cardiovasc Diagn Ther 2017, 7 (3), 252-263; Keyserling, et al. Clin Drug Investig 2017, 37 (5), 483-491 ).

[0066] Spherical HDL species are most correlated with reduced ASCVD (Mendivil, et al. Arterioscler Thromb Vase Biol 2016, 36 (1 ), 156-65). However, these HDLs are more difficult to synthesize due to the enzymatic steps required to mature discoidal HDLs into cholesterol- and cholesteryl ester-containing spherical HDLs. Previous studies have attempted to circumvent these biological maturation steps by using inorganic nanoparticles as templates (Thaxton et al. J. Am. Chem. Soc. 2009, 131, 1384-1385; Skajaa et al., Arterioscler Thromb Vase Biol 2010, 30 (2), 169-76; Cormode, et al. Nano Lett 2008, 8 (11), 3715-23). The templates can be functionalized with phospholipids and apoA-l, and successfully restrict the size of HDL mimics to < 15 nm diameter (Forte et al., Methods Enzymol 1986, 128, 442-57).However, these materials fail to recapitulate a critical feature of native spherical HDL, namely a robust, soft material core capable of dynamically loading and off-loading cholesterol and cholesteryl esters. Apart from inorganic template materials, soft-core discoidal HDL-like particles have also been synthesized using amphiphilic peptides (Zhao et al. Journal of the American Chemical Society 2013, 135 (36), 13414-24).

[0067] Synthesis of certain HDL NP has been described in US Patent Publication No. 20220151928, herein incorporated by reference, and Thaxton et al., J. Am. Chem. Soc. 2019, 141 , 9753-9757.

[0068] As used herein, the terms “HDL-like”, “HDL-mimetic”, and “HDL mimic” are used interchangeably to refer to a synthetic HDL-NP.

[0069] As used herein, “dendritic lipid scaffold” refers to a scaffold comprising tunable hydrophilic, hydrophobic or amphiphilic molecule cores that can be used to facilitate 3-D supramolecular assembly of other core and / or shell lipids to form lipid nano- and microparticles. Tunable refers to the ability of the nanoparticles to be designed, or tuned, to a size or negative charge range depending on the organic scaffold used in the particle. Tunability provides advantages for therapeutics uses and targeting to different cells or tissues.

[0070] The HDL-NP has an organic scaffold. An organic scaffold as used herein refers to non-metallic material, soft-core, having a 3 dimensional structure and charge sufficient to organize and hold a lipid layer in a spherical shape. A “spherical” shape or structure herein refers to a structure having a round or sphere-like structure. The structure does not need to be perfectly round or an exact sphere, but rather is an approximate sphere shape.

[0071] In some aspects, the present disclosure provides a spherical lipoprotein-like nanoparticle (HDL-NP) comprising: a core and a shell surrounding and attached to the core, wherein the core comprises a lipid conjugated organic scaffold. The shell may be a lipid shell that is linked to the components of the organic scaffold at one or more points. In some embodiments, the shell is a lipid bilayer or monolayer.

[0072] In various embodiments, a structure described herein or a portion thereof, such as a shell of a structure, includes one or more natural or synthetic lipids or lipid analogs (i.e., lipophilic molecules). One or more lipids and / or lipid analogues may form a single layer or a multi-layer (e.g., a bilayer) of a structure. In some instances where multi-layers are formed, the natural or synthetic lipids or lipid analogs interdigitate (e.g., between different layers). Non-limiting examples of natural or synthetic lipids or lipid analogs include fatty acyls, glycerolipids, glycerophospholipids, sphingolipids, saccharolipids and polyketides (derived from condensation of ketoacyl subunits), and sterol lipids and prenol lipids (derived from condensation of isoprene subunits).

[0073] In various embodiments, a structure described herein includes one or more phospholipids, e.g., having a C12, C14, C16, C18 or C20 fatty acid or lipid moiety. The one or more lipids may include, for example, phosphatidylcholine, phosphatidylglycerol, lecithin, P, y-dipalmitoyl-a-lecithin, sphingomyelin, phosphatidylserine, phosphatidic acid, N-(2,3-di(9- (Z)-octadecenyloxy))-prop-1-yl-N,N,N-trimethylammonium chloride, phosphatidylethanolamine, lysolecithin, lysophosphatidylethanolamine, phosphatidylinositol, cephalin, cardiolipin, cerebrosides, dicetylphosphate, dioleoylphosphatidylcholine, dipalmitoylphosphatidylcholine, dipalmitoylphosphatidylglycerol, dioleoylphosphatidylglycerol, palmitoyl-oleoyl-phosphatidylcholine, di-stearoyl- phosphatidylcholine, stearoyl-palmitoyl-phosphatidylcholine, di-palmitoyl- phosphatidylethanolamine, di-stearoyl-phosphatidylethanolamine, di-myrstoyl- phosphatidylserine, di-oleyl-phosphatidylcholine, 1 ,2-dipalmitoyl-sn-glycero-3- phosphothioethanol, 1 ,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC), 1 ,2-dioleoyl-sn- glycero-3-phosphocholine (DOPC), 1 ,2-diolinoleoyl-sn-glycero-3-phosphocholine (DLPC), 1 ,2-dioleoyl-sn-glycero-3-phospho-L-serine (DOPS), 1 ,2-distearoyl-sn-glycero-3-phospho- (1 '-rac-glycerol) (18:0 PG), or 1 ,2-dilinoleoyl-sn-glycero-3-phospho-(1'-rac-glycerol) (18:2 PG), and combinations thereof. In some cases, a shell (e.g., a bilayer) of a structure includes 50-200 natural or synthetic lipids or lipid analogs (e.g., phospholipids). For example, the shell may include less than about 500, less than about 400, less than about 300, less than about 200, or less than about 100 natural or synthetic lipids or lipid analogs (e.g., phospholipids), e.g., depending on the size of the structure.

[0074] Non-phosphorus containing lipids may also be used such as stearylamine, docecylamine, acetyl palmitate, and fatty acid amides. In other embodiments, other lipids such as fats, oils, waxes, cholesterol, sterols, fat-soluble vitamins (e.g., vitamins A, D, E and K), glycerides (e.g., monoglycerides, diglycerides, triglycerides) can be used to form portions of a structure described herein.

[0075] The lipid components of the organic core may be selected from the group consisting of lauric, myristic, palmitic, stearic, and arachidic acids and / or their unsaturated derivatives (oleic, linoleic, linolenic, eleostearic, and arachidonic acids), which can be mono- or polyunsaturated.

[0076] A portion of a structure described herein such as a shell or a surface of a nanostructure may optionally include one or more alkyl groups, e.g., an alkane-, alkene-, or alkyne-containing species, that optionally imparts hydrophobicity to the structure. An “alkyl” group refers to a saturated aliphatic group, including a straight-chain alkyl group, branched- chain alkyl group, cycloalkyl (alicyclic) group, alkyl substituted cycloalkyl group, and cycloalkyl substituted alkyl group. The alkyl group may have various carbon numbers, e.g.,between C2 and C40, and in some embodiments may be greater than C5, C10, C15, C20, C25, C30, or C35. In some embodiments, a straight chain or branched chain alkyl may have 30 or fewer carbon atoms in its backbone, and, in some cases, 20 or fewer. In some embodiments, a straight chain or branched chain alkyl may have 12 or fewer carbon atoms in its backbone (e.g., C1 -C12 for straight chain, C3-C12 for branched chain), 6 or fewer, or 4 or fewer. Likewise, cycloalkyls may have from 3-10 carbon atoms in their ring structure, or 5, 6 or 7 carbons in the ring structure. Examples of alkyl groups include, but are not limited to, methyl, ethyl, propyl, isopropyl, cyclopropyl, butyl, isobutyl, tert-butyl, cyclobutyl, hexyl, cyclochexyl, and the like.

[0077] In some embodiments, the spherical HDL-NPs of the instant disclosure further comprise apolipoprotein. The apolipoprotein can be apolipoprotein A (e.g., apo A-l, apo A-ll, apo A-IV, and apo A-V), apolipoprotein B (e.g., apo B48 and apo B100), apolipoprotein C (e.g., apo C-l, apo C-ll, apo C-lll, and apo C-IV), and apolipoproteins D, E, and H. Specifically, apo A1 , apo A2 , and apo E promote transfer of cholesterol and cholesteryl esters to the liver for metabolism and may be useful to include in structures described herein. Additionally, or alternatively, a structure described herein may include one or more peptide analogues of an apolipoprotein, such as one described above. Of course, other proteins (e.g., non-apolipoproteins) can also be included in the nanostructures described herein. In some embodiments, the apolipoprotein is apolipoprotein A-l.

[0078] In some embodiments, the organic scaffold comprising a small molecule with a plurality of terminal functional groups is coupled to one or more lipid to form the dendritic lipid scaffold. The hydrophobic small molecule-lipid conjugate comprises any small molecule capable of being linked to a lipid. In some embodiments, the small molecule is pentaerythritol, sorbitol or tetraA7s(4-functionalized phenyl)methane.

[0079] The small molecule may be linked directly to the lipid or may be linked through the use of a functional group. The functional group may include any suitable end group that can be used to functionalize the lipid to the small molecule, e.g., an amino group (e.g., an unsubstituted or substituted amine), an amide group, an azide, an imine group, a carboxyl group, or a sulfate group. In some instances, the functional group includes at least a second end group. In other embodiments, the second end group may be a reactive group that can covalently attach to another functional group. In some embodiments, the lipid is coupled to the small molecule with a plurality of terminal functional groups. In various embodiments, the plurality of functional groups are hydroxyl groups. In various embodiments, the plurality of functional groups are amide groups. In various embodiments, the functional groups are carboxylates. In these cases, the fatty acids can be reduced to the corresponding alcohols,e.g., lauryl, stearyl, and oleyl alcohols, and coupled to the small molecule core to form the dendritic lipid scaffold.

[0080] In various embodiments, components of the shell are selected from the group consisting of 1 ,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC), 1 ,2-dioleoyl-sn-glycero-3- phosphocholine (DOPC), 1 ,2-diolinoleoyl-sn-glycero-3-phosphocholine (DLPC), 1 ,2-dioleoyl- sn-glycero-3-phospho-L-serine (DOPS), 1 ,2-distearoyl-sn-glycero-3-phospho-(1'-rac- glycerol) (18:0 PG), or 1 ,2-dilinoleoyl-sn-glycero-3-phospho-(1 '-rac-glycerol) (18:2 PG).

[0081] In various embodiments, the organic core HDL nanoparticles (oc-HDL NP) provided herein include DPPC Polyethylene glycol) sorbitol hexaoleate (Sorb(PEG)l -Oe), DPPC Polyethylene glycol) sorbitol hexastearate (Sorb(PEG)l-Se), DPPC PE-S4, DPPC PE-O4, DOPC PE-O4 and DOPS PE-O4.

[0082] In various embodiments, the fatty acid tails of the organic cores are replaced with phospholipid tails.

[0083] In various embodiments, components of the shell are selected from the group consisting of ionizable derivatives of lauric, myristic, palmitic, stearic, and arachidic acids and / or their unsaturated derivatives (oleic, linoleic, linolenic, eleostearic, and arachidonic acids), which can be mono- or polyunsaturated. Such derivatives can be salts of the fatty acids or the acid themselves being ionized in situ and assembled under the appropriate conditions.

[0084] In some embodiments, the spherical HDL-NP further comprises a therapeutic agent linked to the HDL-NP. In some embodiments, the therapeutic agent is a therapeutic nucleic acid. In some embodiments, the therapeutic agent is an anti-cancer agent. In some embodiments, the anti-cancer agent is chemotherapeutic agent. In some embodiments, the therapeutic agent is an anti-oxidant. In some embodiments, the therapeutic agent is an antiinflammatory agent.

[0085] A therapeutic nucleic acid may include any nucleic acid such as but not limited to a polynucleotide, a DNA sequence, a DNA sequence encoding a therapeutic protein, an RNA sequence, a small interfering RNA (siRNA), mRNA, a short-hairpin RNA (shRNA), a micro RNA (miRNA), an antisense oligonucleotide, a triplex DNA, a plasmid DNA (pDNA) or any combinations thereof. In some embodiments, a therapeutic nucleic acid may be treated or chemically modified. For example, a therapeutic nucleic acid may contain inter-nucleotide linkages other than phosphodiester bonds, such as phosphorothioate, methylphosphonate, methylphosphodiester, phosphorodithioate, phosphoramidate, phosphotriester, or phosphate ester linkages, which in some embodiments may confer increased stability. Nucleic acid stability may also be increased by incorporating 3'-deoxythymidine or 2'-substitutednucleotides (substituted with, e.g., an alkyl group) into the nucleic acid during synthesis or by providing the nucleic acid as phenylisourea derivatives, or by having other molecules, such as aminoacridine or poly-lysine, linked to the 3' end of the nucleic acid. Modifications of a RNA and / or a DNA may be present throughout the oligonucleotide or in selected regions of the nucleic acid, e.g., the 5' and / or 3' ends, for example by methylation.

[0086] In certain embodiments, the anti-cancer agent is a chemotherapeutic drug such as Paclitaxel, Cisplatin, Carboplatin, Topotican and Doxoribicin.

[0087] The size, physical properties and functional properties of the HDL-NP of the instant disclosure are similar to that of naturally occurring HDL-NP and distinct from other synthetic HDL-NP. These properties include, for example, spherical shape, surface chemistry, size, hydrodynamic diameter, zeta potential, cholesterol efflux, cholesterol delivery, and therapeutic functions such as suppression of inflammation.

[0088] In some embodiments, the spherical HDL-NP of the instant disclosure, can be assessed based on the hydrodynamic diameter. The hydrodynamic diameter of the spherical HDL-NP is similar to that of naturally occurring HDL-NP and distinct from other synthetic HDL-NP. Hydrodynamic diameter assesses the size of a hypothetical hard sphere that diffuses in the same manner as that of the particle being measured and provides an indication of the diffusional properties of the particle that will be indicative of the apparent size of the dynamic hydrated / solvated particle. It may be measured by Dynamic Light Scattering (DLS). In some embodiments the hydrodynamic diameter is greater than 8 nm e.g., about 8 nm to about 30 nm. In some embodiments, the hydrodynamic diameter is 8 nm-17 nm. In some embodiments, the hydrodynamic diameter is 10 nm-15 nm. In some embodiments, the hydrodynamic diameter is 12 nm-14 nm.

[0089] The spherical HDL-like nanoparticles of the present disclosure can have a diameter with a largest cross-sectional dimension (or, sometimes, a smallest cross-section dimension) of, for example, less than or equal to about 500 nm, less than or equal to about 250 nm, less than or equal to about 100 nm, less than or equal to about 75 nm, less than or equal to about 50 nm, less than or equal to about 40 nm, less than or equal to about 35 nm, less than or equal to about 30 nm, less than or equal to about 25 nm, less than or equal to about 20 nm, less than or equal to about 15 nm, or less than or equal to about 5 nm. In some embodiments the spherical HDL-NP has a diameter of about 5-60 nm, 10-50 nm, 5-40 nm, 10-30 nm, 10-20 nm, 5-30 nm, 5-25 nm, 5-20 nm, 5-15 nm, 5-10 nm, 8-15 nm, 8-14 nm, 8-13 nm, 8-12 nm, 8-11 nm, 8-10 nm, or 10-12 nm.

[0090] The lipoprotein-like nanoparticles of the present disclosure can have a core with a largest cross-sectional dimension (or, sometimes, a smallest cross-section dimension) of, forexample, less than or equal to about 300 nm, less than or equal to about 250 nm, less than or equal to about 100 nm, less than or equal to about 75 nm, less than or equal to about 50 nm, less than or equal to about 40 nm, less than or equal to about 35 nm, less than or equal to about 30 nm, less than or equal to about 25 nm, less than or equal to about 20 nm, less than or equal to about 15 nm, or less than or equal to about 5 nm. In some cases, the core has an aspect ratio of greater than about 1 :1 , greater than 3:1 , or greater than 5:1 . As used herein, “aspect ratio” refers to the ratio of a length or a width, where length and width are measured perpendicular to one another, and the length refers to the longest linearly measured dimension.

[0091] In some embodiments, the shell has a negative zeta potential similar to or more negative than human HDL. In some embodiments, the spherical HDL-NP has a zeta potential of about -5 mV to -25 mV or -10 to -30 mV. In some embodiments, the zeta potential of the HDL-like nanoparticles is about -20 or about -25 millivolts (mV). In some embodiments, the zeta potential of the HDL-like nanoparticles is greater than -20 mV. In some embodiments, the zeta potential is less than -20 mV. In some embodiments, the zeta potential of the HDL-like nanoparticles is selected from a group consisting of about -5, mV, - 6 mV, -7 mV, -8 mV, -9 mV, -10 mV, -11 mV, -12 mV, -13 mV, -14 mV, -15 mV, -16 mV, -17 mV, -18 mV, -19 mV, -20 mV, -21 mV, -22 mV, -23 mV, -24 mV, -25 mV, -26 mV, -27 mV, - 28 mV, -29 mV and -30 mV. In some embodiments, the zeta potential is that of human HDL. Zeta potential may be assessed using methods known in the art, including the methods disclosed herein.

[0092] In one embodiment, the HDL-NP organic core comprises PE-S4 and has a diameter of about 10 nm to about 20 nm and a zeta potential between about -10 mV to about -20 mV.

[0093] In one embodiment, the HDL-NP organic core comprises PE-O4 and has a diameter of about 8 nm to about 17 nm and a zeta potential between about -5 mV to about - 15 mV.

[0094] In one embodiment, the HDL-NP organic core comprises Sorb(PEG)n-Se and has a diameter of about 10 nm to about 30 nm and a zeta potential between about -10 mV to about -20 mV.

[0095] In one embodiment, the HDL-NP organic core comprises Sorb(PEG)n-Oe and has a diameter of about 10 nm to about 25 nm and a zeta potential between about -15 mV to about -25 mV.

[0096] In one embodiment, the HDL-NP organic core comprises S4 and has a diameter of about 7 nm to about 25 nm and a zeta potential between about -10 mV to about -20 mV.

[0097] In one embodiment, the HDL-NP organic core comprises O4 and has a diameter of about 7 nm to about 25 nm and a zeta potential between about -10 mV to about -20 mV.

[0098] In some embodiments, the lipoprotein-like nanoparticles of the present disclosure do not include a peptide-based scaffold material.

[0099] The lipoprotein-like nanoparticles (also referred to as synthetic particles or synthetic HDL particle, HDL-NP or HDL-like NP) of the present disclosure are capable of executing salient HDL functions including cholesterol efflux from macrophages (or from lipid- laden macrophages), cholesterol delivery to hepatocytes, supporting lecithin cholesterol acyltransferase activity, and suppression of inflammation. In various embodiments, the spherical HDL-NP has a more efficient cholesterol transport capacity than that of apolipoprotein or a synthetic HDL nanoparticle with a gold core. Cholesterol transport capacity may be assessed using in vitro assays such as the assays described herein. The cholesterol transport capacity may be assessed relative to a standard known value of cholesterol transport or relative to a negative control such a synthetic HDL-NP particle having a gold core or APO-A1 or relative to a positive control such as naturally occurring HDL-NP.Methods of Use

[0100] The compositions and methods of the present disclosure can be used in applications including, but not limited to, cancer therapy, inflammatory disease, cardiovascular disease therapy, bone disease therapy, and immune disease therapy.

[0101] Inflammation promoted by macrophage NF-kB activity is a hallmark of ASCVD that drives disease progression and increases morbidity and mortality. Provided herein is a method for treating cardiovascular disease by administering the oc-HDL-NP of the disclosure. Cardiovascular disease is a vascular endothelial cell dysfunction and certain symptoms begin, including as conventional or above the heart and vascular system-on, atherosclerosis, hypertension, gojihyeol, coronary heart disease (heart attack), cerebrovascular diseases (stroke, dementia), peripheral vascular disease, arrhythmia, heart failure, congestive heart disease, cardiac disease and for at least the name of the heart and blood vessels, including, but not limited thereto. Some of the main factors of cardiovascular disease include, but they are not limited to expression of genetic factors, lifestyle habits, and complications of diabetes.

[0102] As used herein cardiovascular diseases include, but are not limited to, arteriosclerosis, coronary heart disease, ischemia, endothelium dysfunction, in particular those dysfunctions affecting blood vessel elasticity, restenosis, thrombosis, angina, high blood pressure, cardiomyopathy, hypertensive heart disease, heart failure, corpulmonale,cardiac dysrhythmias, endocarditis, inflammatory cardiomegaly, myocarditis, myocardial infarction, valvular heart disease, stroke and cerebrovascular disease, aortic valve stenosis, congestive heart failure, and peripheral arterial disease.

[0103] In some embodiments, the compositions of the disclosure restore and / or improve cardiovascular parameters to normal ranges in a subject diagnosed with or at risk of a cardiovascular disease. Normal ranges of cardiovascular parameters include but are not limited to, an end-diastolic volume (EDV) from about 65-240 mL, an end-systolic volume (ESV) from about 16-143m L, a stroke volume from about 55-100 mL, an ejection fraction from about 55-70%, a heart rate from about 60-100 bpm, and / or cardiac output of about 4.0- 8.0 L / min.

[0104] In other aspects a method of treating an inflammatory disorder by administering to a subject having an inflammatory disorder a spherical lipoprotein-like nanoparticle (HDL-NP) comprising: a core and a shell surrounding and attached to the core, wherein the core comprises a lipid conjugated organic scaffold as described herein in an effective amount to treat the inflammatory disorder is provided.

[0105] The compositions of the disclosure may also be used to treat inflammatory diseases. Examples of inflammatory diseases include, but are not limited to, acne vulgaris, asthma, autoimmune diseases (e.g., acute disseminated encephalomyelitis (ADEM), Addison's disease, agammaglbulinemia, alopecia areata, amyotrophic lateral sclerosis, ankylosing spondylitis, antiphospholipid syndrome, antisynthetase syndrome, atopic allergy, atopic dermatitis, autoimmune aplastic anemia, autoimmune cardiomyopathy, autoimmune enteropathy, autoimmunehemolytic anemia, autoimmune hepatitis, autoimmune inner ear disease, autoimmune lymphoproliferative syndrome, autoimmune peripheral neuropathy, autoimmune pancreatitis, autoimmune polyendocrine syndrome, autoimmune progesterone dermatitis, autoimmune thrombocytopenic purpura, autoimmune urticaria, autoimmune uveitis, Balo concentric sclerosis, Bechet’s disease, Berger's disease, Bickerstaff's encephalitis, Blau syndrome, bullous pemphigoid, Castleman's disease, celiac disease, Chagas disease, chronic inflammatory demyelinating polyneuropathy, chronic recurrent multifocal osteomyelitis, chronic obstructive pulmonary disease, Churg-Strauss syndrome, cicatricial pemphigoid, Cogan syndrome, cold agglutinin disease, complement component 2 deficiency, contact dermatitis, cranial arteritis, CREST syndrome, Crohn's disease, Cushing's syndrome, cutaneous leukocytoclastic vasculitis, Dego's disease, Dercum's disease, dermatitis herpetiformis, dermatomyositis, diabetes mellitus type 1 , diffuse cutaneous systemic sclerosis, Dressier's syndrome, drug-induced lupus, discoid lupus erythematosus, eczema, endometriosis, enthesitis-related arthritis, eosinophilic fasciitis, eosinophilic gastroenteritis, epidermolysis bullosa acquisita, erythema nodosum,erythroblastosis fetalis, essential mixed cryoglobulinemia, Evan's syndrome, fibrodysplasia ossificans progressive, fibrosing alveolitis, gastritis, gastrointestinal pemphigoid, giant cell arteritis, glomerulonephritis, Goodpasture's syndrome, Grave's disease, Guillain-Barre syndrome, Hashimoto's encephalopathy, Hashimoto's thyroiditis, Henoch-Schonlein purpura, herpes gestationis, hidradenitis suppurativa, Hughes-Stovin syndrome, hypogammaglobulinemia, idiopathic inflammatory demyelinating diseases, idiopathic pulmonary fibrosis, idiopathic thrombocytopenic purpura, IgA nephropathy, inclusion body myositis, chronic inflammatory demyelinating polyneuropathy, interstitial cystitis, juvenile idiopathic arthritis, Kawasaki's disease, Lambert-Eaton myasthenic syndrome, leukocytoclastic vasculitis, lichen planus, lichen sclerosus, linear IgA disease, lupus erythematosus, Majeed syndrome, Meniere's disease, microscopic polyangiitis, mixed connective tissue disease, morphea, Mucha-Habermann disease, myasthenia gravis, myositis, narcolepsy, neuromyelitis optica, neuromyotonia, ocular cicatricial pemphigoid, opsoclonus myoclonus syndrome, Ord's thyroiditis, palindromic rheumatism, PANDAS, paraneoplastic cerebellar degeneration, paroxysmal nocturnal hemoglobinuria, Parry Romberg syndrome, Parsonage-Turner syndrome, pars planitis, pemphigus vulgaris, pernicious anemia, perivenous encephalomyelitis, POEMS syndrome, polyarteritis nodosa, polymyalgia rheumatic, polymyositis, primary biliary cirrhosis, primary sclerosing cholangitis, progressive inflammatory neuropathy, psoriatic arthritis, pyoderma gangrenosum, pure red cell aplasia, Rasmussen's encephalitis, raynaud phenomenon, relapsing polychondritis, Reiter's syndrome, restless leg syndrome, retroperitoneal fibrosis, rheumatic fever, Schnitzler syndrome, scleritis, scleroderma, serum sickness, Sjogren's syndrome, spondyloarthropathy, stiff person syndrome, subacute bacterial endocarditis, Susac's syndrome, Sweet's syndrome, sympathetic ophthalmia, Takayasu's arteritis, temporal arteritis, thrombocytopenia, Tolosa-Hunt syndrome, transverse myelitis, ulcerative colitis, undifferentiated connective tissue disease, undifferentiated spondyloarthropathy, vitiligo, and Wegener's granulomatosis), celiac disease, chronic prostatitis, glomerulonephritis, hypersensitivities, inflammatory bowel diseases, pelvic inflammatory disease, reperfusion injury, rheumatoid arthritis, sarcoidosis, transplant rejection, vasculitis, interstitial cystitis, and osteoarthritis.

[0106] In other aspects the disclosure contemplates a method for treating a cancer, comprising administering to a subject having a cancer a spherical lipoprotein-like nanoparticle (HDL-NP) as described herein in an amount effective to treat the cancer.

[0107] Cancers are generally characterized by unregulated cell growth, formation of malignant tumors, and invasion to nearby parts of the body. Cancers may also spread to more distant parts of the body through the lymphatic system or bloodstream. Cancers maybe a result of gene damage due to tobacco use, certain infections, radiation, lack of physical activity, obesity, and / or environmental pollutants. Cancers may also be a result of existing genetic faults within cells to cause diseases due to genetic heredity. Screenings may be used to detect cancers before any noticeable symptoms appear and treatment may be given to those who are at higher risks of developing cancers (e.g., people with a family history of cancers). Examples of screening techniques for cancer include but are not limited to physical examination, blood or urine tests, medical imaging, and / or genetic testing. Non-limiting examples of cancers include: bladder cancer, breast cancer, colon and rectal cancer, endometrial cancer, kidney or renal cell cancer, leukemia, lung cancer, melanoma, NonHodgkin lymphoma, pancreatic cancer, prostate cancer, ovarian cancer, stomach cancer, wasting disease, and thyroid cancer.

[0108] In some embodiments, the nanoparticles described herein are useful for treating a cancer that expresses or overexpresses scavenger receptor class B type I (SR-BI). Nonlimiting examples of cancers that express or overexpress SR-BI include human prostate cancer, breast cancer, and renal cell carcinoma. Additional non-limiting examples of cancers and cancer cell lines that overexpress SR-BI are listed in Rajora et al. Front Pharmacol. (2016) 7:326. As described herein, the term “overexpression” or “increased expression,” refers to an increased level of expression of a given gene product in a given cell, cell type or cell state, as compared to a reference cell, for example, a non-cancer cell or a cancer cell that does not overexpress SR-BI. In some embodiments the cancer cell expresses any level of SR-BI.

[0109] The nanoparticles are also useful for treating and preventing autoimmune disease or disorder. Autoimmune disease or disorder is a class of diseases in which a subject’s own antibodies react with host tissue or in which immune effector T cells are autoreactive to endogenous self peptides and cause destruction of tissue. Thus, an immune response is mounted against a subject’s own antigens, referred to as self antigens. Autoimmune diseases or disorders include, but are not limited to, rheumatoid arthritis, Crohn's disease, multiple sclerosis, systemic lupus erythematosus (SLE), autoimmune encephalomyelitis, myasthenia gravis (MG), Hashimoto’s thyroiditis, Goodpasture’s syndrome, pemphigus (e.g., pemphigus vulgaris), Grave’s disease, autoimmune hemolytic anemia, autoimmune thrombocytopenic purpura, scleroderma with anti-collagen antibodies, mixed connective tissue disease, polymyositis, pernicious anemia, idiopathic Addison’s disease, autoimmune- associated infertility, glomerulonephritis (e.g., crescentic glomerulonephritis, proliferative glomerulonephritis), bullous pemphigoid, Sjogren’s syndrome, insulin resistance, and autoimmune diabetes mellitus.

[0110] The compositions containing an effective amount of the HDL-NP described herein can be administered for prophylactic or therapeutic treatments. In prophylactic applications, compositions can be administered to a patient with a clinically determined predisposition or increased susceptibility to development of a disorder associated with elevated levels of cholesterol, cardiovascular diseases, hyperproliferative diseases (e.g., cancer), inflammatory diseases, dyslipidemia, and other pathological conditions associated with cholesterol. Compositions of the invention can be administered to the patient (e.g., a human) in an amount sufficient to delay, reduce, or preferably prevent the onset of the clinical disease. In therapeutic applications, compositions are administered to a patient (e.g., a human) already suffering from a cardiovascular disease, hyperproliferative diseases (e.g., cancer), an inflammatory disease, and other pathological conditions associated with cholesterol, in an amount sufficient to cure or at least partially arrest the symptoms of the condition and its complications. An amount adequate to accomplish this purpose is defined as a "therapeutically effective dose," an amount of a compound sufficient to substantially improve some symptom associated with a disease or a medical condition. A therapeutically effective amount of an agent or composition is not required to cure a disease or condition but will provide a treatment for a disease or condition such that the onset of the disease or condition is delayed, hindered, or prevented, or the disease or condition symptoms are ameliorated, or the term of the disease or condition is changed or, for example, is less severe or recovery is accelerated in an individual.

[0111] In various embodiments, the disclosure relates to a method of reducing NF-kB and / or TNF-a activity, comprising administering to a subject an effective amount of a composition comprising any of the spherical HDL-NP of the disclosure. In various embodiments, the NF-kB and / or TNF-a activity is reduced in a subject having cancer, an inflammatory disease, a cardiovascular disease, a bone disease, or an immune disease.

[0112] As used herein, a “subject” or a “patient” refers to any mammal (e.g., a human). Examples of subjects or patients include a human, a non-human primate, a cow, a horse, a pig, a sheep, a goat, a dog, a cat or a rodent such as a mouse, a rat, a hamster, or a guinea pig. Generally, the invention is directed toward use with humans.Pharmaceutical Compositions

[0113] In some embodiments, the lipoprotein-like nanoparticles of the present disclosure are in a pharmaceutical composition. The “pharmaceutical composition” or “pharmaceutically acceptable” composition may comprise a therapeutically effective amount of one or more of the structures described herein, formulated together with one or more pharmaceutically acceptable carriers, additives, and / or diluents. The pharmaceutical compositions describedherein may be useful for treating sepsis or other related diseases. It should be understood that any suitable structures described herein can be used in such pharmaceutical compositions, including those described in connection with the figures.

[0114] The pharmaceutical compositions may be formulated for administration in solid or liquid form, including those adapted for the following: oral administration, for example, drenches (aqueous or non-aqueous solutions or suspensions), tablets, e.g., those targeted for buccal, sublingual, and systemic absorption, boluses, powders, granules, pastes for application to the tongue; parenteral administration, for example, by subcutaneous, intramuscular, intravenous or epidural injection as, for example, a sterile solution or suspension, or sustained-release formulation; topical application, for example, as a cream, ointment, or a controlled-release patch or spray applied to the skin, lungs, or oral cavity; intravaginally or intrarectally, for example, as a pessary, cream or foam; sublingually; ocularly; transdermally; or nasally, pulmonary and to other mucosal surfaces.

[0115] The phrase “pharmaceutically acceptable” is employed herein to refer to those structures, 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.

[0116] The phrase “pharmaceutically-acceptable carrier” as used herein means a pharmaceutically-acceptable material, composition or vehicle, such as a liquid or solid filler, diluent, excipient, or solvent encapsulating material, involved in carrying or transporting the subject compound from one organ, or portion of the body, to another organ, or portion of the body. Each carrier must be “acceptable” in the sense of being compatible with the other ingredients of the formulation and not injurious to the patient. Some examples of materials which can serve as pharmaceutically-acceptable carriers include: sugars, such as lactose, glucose and sucrose; starches, such as corn starch and potato starch; cellulose, and its derivatives, such as sodium carboxymethyl cellulose, ethyl cellulose and cellulose acetate; powdered tragacanth; malt; gelatin; talc; excipients, such as cocoa butter and suppository waxes; oils, such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil and soybean oil; glycols, such as propylene glycol; polyols, such as glycerin, sorbitol, mannitol and polyethylene glycol; esters, such as ethyl oleate and ethyl laurate; agar; buffering agents, such as magnesium hydroxide and aluminum hydroxide; alginic acid; pyrogen-free water; isotonic saline; Ringer's solution; ethyl alcohol; pH buffered solutions; polyesters, polycarbonates and / or polyanhydrides; and other non-toxic compatible substances employed in pharmaceutical formulations.

[0117] Wetting agents, emulsifiers and lubricants, such as sodium lauryl sulfate and magnesium stearate, as well as coloring agents, release agents, coating agents, sweetening, flavoring and perfuming agents, preservatives and antioxidants can also be present in the compositions.

[0118] Examples of pharmaceutically-acceptable antioxidants include: water soluble antioxidants, such as ascorbic acid, cysteine hydrochloride, sodium bisulfate, sodium metabisulfite, sodium sulfite and the like; oil-soluble antioxidants, such as ascorbyl palmitate, butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT), lecithin, propyl gallate, alpha-tocopherol, and the like; and metal chelating agents, such as citric acid, ethylenediamine tetraacetic acid (EDTA), sorbitol, tartaric acid, phosphoric acid, and the like.

[0119] The structures described herein may be orally administered, parenterally administered, subcutaneously administered, and / or intravenously administered. In certain embodiments, a structure or pharmaceutical preparation is administered orally. In other embodiments, the structure or pharmaceutical preparation is administered intravenously. Alternative routes of administration include sublingual, intramuscular, and transdermal administrations.Methods of Making

[0120] In one aspect, the disclosure relates to a method for making a spherical lipoprotein-like nanoparticle (HDL-NP) comprising: preparing an organic scaffold solution; contacting the organic scaffold solution with apolipoprotein to generate an organic scaffold- Apo solution; contacting the organic scaffold-Apo solution with a lipid solution to generate an organic scaffold-Apo-lipid solution; admixing the organic scaffold-Apo-lipid solution by vortexing and / or sonication to form HDL-NP; isolating the HDL-NP.

[0121] It is contemplated that the method makes HDL-NP comprising a core and a shell surrounding and attached to the core, wherein the core comprises a lipid conjugated organic scaffold. The organic scaffold solution comprises any one of pentaerythritol distearate (PE- 82), pentaerythritol dioleate (PE-O2), pentaerythritol tetrastearate (PE-S4), pentaerythritol tetraoleate (PE-O4), sorbitol(polyethylene glycol)n-S6 (Sorb(PEG)n-Se) or sorbitol(polyethylene glycol)n-06 (Sorb(PEG)n-Oe) tetrastearate (S4) or tetraoleate (O4).

[0122] The organic scaffold optionally comprises a headgroup-modified fatty acid. The headgroup-modified fatty acid can comprise C12, C14, C16, C18 or C20 tails, and is monounsaturated or polyunsaturated.

[0123] In some embodiments, the lipid components of the organic scaffold are selected from the group consisting of lauric acid, myristic acid, palmitic acid, stearic acid, arachidic acid, oleic acid, linoleic acid, linolenic acid, eleostearic acid, and arachidonic acid.

[0124] In various embodiments, the shell comprises components selected from the group consisting of 1 ,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC), 1 ,2-dioleoyl-sn-glycero-3- phosphocholine (DOPC), 1 ,2-diolinoleoyl-sn-glycero-3-phosphocholine (DLPC), 1 ,2-dioleoyl- sn-glycero-3-phospho-L-serine (DOPS), 1 ,2-distearoyl-sn-glycero-3-phospho-(1'-rac- glycerol) (18:0 PG), or 1 ,2-dilinoleoyl-sn-glycero-3-phospho-(1 '-rac-glycerol) (18:2 PG).

[0125] In various embodiments, the organic scaffold comprises a configuration of lipid alcohols coupled with a small molecule to form the oc-scaffold. In various embodiments, the lipids are coupled to tetraA7s(4-functionalized phenyl)methane, e.g., a small molecule with four terminal azides (SM-Az4).

[0126] In various embodiments, the organic scaffold comprises a sorbitol-polyethylene glycol conjugated fatty acid, such as polyethylene glycol sorbitol hexaoleate (Sorb(PEG)i- Oe) or the reduced compound polyethylene glycol sorbitol hexastearate Sorb(PEG)i-Se.

[0127] In some aspects of the method, to form HDL nanoparticles, the organic scaffolds are then reacted with solutions of organic lipid and apolipoprotein to form the HDL nanoparticle. In one embodiment, the core scaffolds are contacted with apolipoprotein followed by addition of the phospholipid group (e.g., DPPC) and admixed to form a suspension. The suspension is then subject to vortexing, sonication and / or stirring to form the nanoparticles having a shell and core. The particles are optionally concentrated, e.g., by centrifugation and filtration, and isolated after a final filtration step.

[0128] In various embodiments, the apolipoprotein is apolipoprotein A (e.g., apo A-l, apo A-ll, apo A-IV, and apo A-V), apolipoprotein B (e.g., apo B48 and apo B100), apolipoprotein C (e.g., apo C-l, apo C-ll, apo C-lll, and apo C-IV), apolipoproteins D, apolipoproteins E or apolipoprotein H.

[0129] The following examples are intended to illustrate but not limit the invention. While they are typical of those that might be used, other procedures known to those skilled in the art may alternatively be used.EXAMPLESEXAMPLE 1 -MATERIALS AND METHODS

[0130] Unless otherwise stated, all reagents and reagent-grade solvents were purchased from Sigma-Aldrich (Milwaukee, Wl) and used as received.3-(stearoyloxy)-2, 2- bis[(stearoyloxy)methyl]propyl stearate (PE-S4) and Polyethylene glycol) sorbitol hexaoleate (Sorb(PEG)i-Oe) cores were acquired from Sigma-Aldrich (Milwaukee, Wl). 1 ,2-dipalmitoyl- sn-glycero-3-phosphocholine (DPPC) was obtained from Avanti Polar Lipids (Alabaster, AL). Cholesteryl lineolate was purchased from Sigma Aldrich. Purified monomeric human apolipoprotein A-l (apoA-l) was obtained from MyBioSource (San Diego, CA). Ultrapure deionized (DI) H2O (18.2 MQ cm resistivity) was obtained from a Millipore system (Milli-Q Biocel).

[0131] Vortexing was carried out on an analog vortex mixer (VWR, Radnor, PA). Sonication was carried out in a water-filled Branson 2510 Ultrasonic Sonicator (St. Louis, MO, USA). Centrifugation was carried out in an Eppendorf Centrifuge 5415 R (Eppendorf AG, Hamburg, Germany) equipped with an FA-45-24-11 rotor. All centrifugations were carried out at either 4°C or 25°C on a Eppendorf Centrifuge. Nanoparticle stirring at room temperature (RT) was carried out on a Thermo-Fisher magnetic stir plate set at 600 rpm.

[0132] Particle protein concentration was determined by conducting bicinchoninic acid assays (BCA) purchased from ThermoFisher. Spectroscopic measurements at an absorbance at 562 nm were obtained using a BioTek Synergy 2 microplate reader (Winooski, VT).

[0133] Mass spectrometric data were collected as negative and positive ions using an LC- TOF mass spectrometer (Agilent 6545 Q-TOF LC / MS). Data from Agilent HPLC and Agilent Q-TOF instruments were processed using Agilent MassHunter Workstation software version10.0.

[0134] Transmission electron microscopy and dynamic light scattering measurements: Dynamic light scattering (DLS) and zeta potential measurements were carried out on a Zetasizer Nano ZS (Malvern Instruments, Malvern, UK) equipped with a He-Ne laser (633 nm). TEM imaging was carried out with a Hitachi HD2300 Scanning Transmission Election Microscope (STEM). Carbon-coated copper 300-mesh grids were purchased from Electron Microscopy Services (Hatfield, PA). Uranyl acetate was purchased from Electron Microscopy Sciences. Non-invasive backscatter method (detection at 173 “ scattering angle) was used. Correlation data were fitted, using the method of cumulants, to the logarithm of the correlation function, yielding the diffusion coefficient, D. The hydrodynamic diameters (Dw) of nanoparticles were calculated using D and the Stokes-Einstein equation (DH= kBT / 3TrriD, where kBis the Boltzmann constant, T is the absolute temperature, and is the solvent viscosity ( / ? = 0.8872 cP for water)). The polydispersity index (PDI) of the nanoparticles —represented as 2c / , where b and c are first- and second-order coefficients, respectively, in a polynomial of a semi-log correlation function — was calculated by cumulants analysis. Size distribution of the nanoparticles was obtained by non-negative least squares (NNLS) analysis.

[0135] To visualize the particles, samples undergo negative stain transmission electron microscopy (TEM). For grid preparation, samples were prepared at a concentration of 500 nM-2 pM protein in PBS and dropcast on glow-discharged carbon-coated copper 300-mesh grids (Electron Microscopy Services) for 1 minute. The samples were removed by touching the corner of the grid with a Kimwipe (KimTech). The grids were stained four times with 1- 2% uranyl acetate for 20 sec with the last stain lasting 4 minutes. The grids washed three times with PBS and air dried prior to imaging. Imaging was conducted using a FEI Tecnai Spirit TEM operating at 80 kV.

[0136] Oligomerization assay. To determine the number of Apo-A1 molecules per particle, particles were crosslinked with bis(sulfosuccinidimidyl)suberate (BS3) (Sigma Aldrich), which stabilizes any higher order oligomerization states of Apo-Al in oc-HDL NPs upon assembly. Particles were diluted to 50 pg / mL protein in PBS. BS3 cross-linking agent is then added to oc-HDL NPs for a final BS3 concentration of 2.5 mM, and the reaction was allowed to proceed for 30 min at room temperature. To produce an Apo-Al oligomer ladder, lipid-free pure Apo-Al was dialyzed in PBS, and then subjected to cross-linking with BS3 (0.25 mM) at elevated protein concentration (500 pg / mL) for 4 h at room temperature. 0.5 M Tris base was used to stop the cross-linking reactions (45 mM final).

[0137] After termination of the reaction, samples were prepared for an Apo-A1 immunoblot. BioRad apparatuses were used for gel electrophoresis and protein transfer. Pre-cast 4-20% polyacrylamide gels (MiniProtean TGX, BioRad) were used for separation. Protein transfer to polyvinylidine fluoride (PVDF) membrane was performed using Tris- Glycine buffer with 20 vol % methanol. PVDF membrane was then rinsed in TBS and blocked in 5 wt % nonfat dry milk for 1 h at rt. Primary antibody (rabbit anti-Apo-AI, Abeam) was added to the membrane at 1 :1000 dilution in 5 wt % nonfat dry milk and incubated at 4 °C overnight. The membrane was then washed in TBS-Tween (0.1%) three times for 10 min each. Secondary antibody (goat anti-rabbit HRP, BioRad) was then added at 1 :2000 dilution in 5 wt % nonfat dry milk. The membrane was then washed three times in TBS-Tween for ten minutes each, and then bathed in chemiluminescence substrate solution for 5 min. Chemiluminescent detection and image capture were performed using the Azure Imaging System (Dublin, CA).

[0138] Tandem cholesterol efflux-influx assays'. For efflux experiments, a validated fluorescence-based assay using Top-Fluor-Cholesterol (Avanti Lipids) was used. J774 macrophages were cultured for at least two passages prior to seeding at 150,000 cells per well in 24 well plates in RPMI, 10% fetal bovine serum (FBS), 1% penicillin-streptomycin (PenStrep) on Day 1. The labeling medium consisted of RPMI media with cholesterol and methyl-p-cyclodextrin (CD; Sigma Aldrich), whereby unlabeled (Sigma Aldrich) and TopFluor cholesterol accounted for 80% and 20% of total cholesterol, respectively. Stock solutions of Top-Fluor-Cholesterol and unlabeled cholesterol (Sigma Aldrich) were prepared in chloroform at a concentration of 8 mM and 250 mM, respectively. Samples were aliquoted, evaporated under N2and stored at -80 °C. Stock solutions were resuspended in chloroform at the selected aliquot volume and transferred into a round-bottom glass vial. The volume of cholesterol added was calculated by the total volume of labeling medium needed for the experiment to achieve a concentration of 0.05 mM (Top-Fluor cholesterol) and 0.2 mM (unlabeled cholesterol). The cholesterol was evaporated under N2u and resuspended in RPMI media containing 20 mM of CD. The medium was sonicated in an ultrasound bath for 30 minutes at 37°C and then placed in a 37°C water bath for 3 hours. The medium was stored at 4°C until the following day.

[0139] On Day 2, the labeling medium was sonicated in the ultrasound bath for 30 minutes at 37 °C. The medium was filtered using a 0.22 pm sterile filter and diluted 2-fold in empty RPMI media to yield a final labeling media comprising 0.125 mM total cholesterol (0.025 mM Top-Fluor and 0.1 mM unlabeled cholesterol), and 10 mM CD. Finally, 2 pg / mL Sandoz, an ACAT inhibitor, was added (Sigma-Aldrich, St. Louis, MO) to prevent esterification of the fluorescent cholesterol. Media was aspirated from 24 well plates and 500 pL of Tol-Fluor cholesterol labeling media was added. Labeling was allowed to proceed for 1 hour in the humidified cell culture incubator at 37 °C. After 1 hour, the labeling media was removed, cells were washed twice with warm RPMI media, and equilibration medium (RPMI, 300 pM cAMP, 5 pM Sandoz, 1% BSA) was added to upregulate the canonical cholesterol efflux receptor ABCA1 . Upregulation was allowed to proceed for 18 h prior to the addition of efflux media. Hepatocytes (HepG2 cells) were plated at 100,000 cells / well in DMEM, 10% FBS, 1% PenStrep in 24 well plates.

[0140] On Day 3, efflux samples containing nanoparticles and controls were prepared in serum-free media (MEM, 25 pM HEPES, 1% PenStrep) to reduce non-specific cholesterol efflux by serum cholesterol carriers. Cells were washed once in warm RPMI, and efflux media was added to each well according to the treatment regimen. Efflux was allowed to proceed for 4 h-12h. Immediately after efflux, HepG2 cells were washed twice in PBS and efflux media from J774 macrophages was removed and added directly to the cells. Influxwas allowed to proceed for 4-12 h prior to harvesting. Top-Fluor cholesterol in J774 macrophages was extracted by incubating 300 pL / well of extraction buffer for 1 hour shaking at RT sealed in Parafilm and protected from light. The extraction buffer was removed and transferred to new Eppendorf tubes and stored at 4 °C until the samples were ready to be measured. At the end of the influx incubation, influx media supernatant was removed and centrifuged at 500 g at 4°C for 10 minutes to discard detached cells and cellular debris. The supernatant was transferred to new Eppendorf tubes. HepG2 were subject to the same extraction protocol as described above. An aliquot (100 |_iL) of the cell-free media or cellular lipid solvent fraction was transferred to a 96-well black plate (Corning Costar). The fluorescent intensity was measured using a BioTek Microplate Reader at excitation and emission wavelengths of 485 nm and 515 nm, respectively.

[0141] Efflux percentages were calculated as ratios of fluorescent signal of cell-free media to J774 macrophages belonging to the PBS control. Influx percentages were calculated as ratios of fluorescent signal of HepG2 cells to J774 macrophages belonging to the PBS control.

[0142] Cholesteryl ester loading assays: For the synthesis of cholesteryl lineolate- incorporated oc-HDL NPs, all reagents (DPPC and lipid-conjugate organic cores in dichloromethane (DCM), Apo-Al in PBS) are prepared fresh. Cholesteryl lineolate was dissolved in DCM at a concentration of 1 mM. In a typical assembly, the lipid conjugate core scaffold (1 nmol in DCM) is combined with Apo-A1 (2 nmol from a 1.4 mg / mL suspension in PBS), followed by the addition of DPPC (150 nmol in DCM) and cholesteryl lineolate (50 nmol in DCM). Following addition of all reagents into the glass vials, the nanoparticles were subjected to the same synthesis and purification process as the oc-HDL NPs without cholesteryl lineolate as described above.

[0143] Samples were diluted 5 times in 1X Reaction Buffer prior to quantification with Amplex Red Cholesterol Assay (Thermo Fisher). Amplex Red Cholesterol Assay was performed per manufacturer’s instructions. Briefly, a cholesterol standard curve was prepared by serially diluting cholesterol in kit-provided aqueous buffer from 32 pg / mL to 125 ng / mL and adding 50 pl to a 96-well black bottom microplate. 50 pl of sample were also added to each well of the microplate in duplicate. 50 pl of Amplex Red reaction mix were then added to each well. Microplates were incubated at 37 °C for 30 minutes.

[0144] NF-KB activity assays: THP1 -Dual cells (100,000 cells / well) were plated into 96 well plates and at the same time treated with media containing the appropriate oc-HDL NP with or without 1 ng / mL lipopolysaccharide (LPS; O55:B5) from Sigma-Aldrich (St. Louis, MO). After 24 h, 20 pL of cells and media were transferred to a new 96 well plate and 180pL of QUANTI-Blue Media (InvivoGen; San Diego, CA) was added to each well. Plates were incubated at 37 °C in a humidified 5 vol% CO2 incubator for 1 h. Secreted embryonic alkaline phosphatase (SEAP) levels as a readout of NF-KB activity were determined by reading the absorbance at 655 nm in a Biotek Synergy 2 microplate reader and the relative NF-KB percentages compared to [LPS + vehicle] control were plotted. The data reported represents an average of three measurements.EXAMPLE 2-SYNTHESIS AND CHARACTERIZATION OF OC-HDL NP

[0145] Synthesis of Sorb(PEG)1-S6 cores

[0146] Prior to the reaction, the commercially available Sorb(PEG)i-Oewas passed through a short pad of silica gel to remove the trace amount of colored impurities and oleic acid. Sorb(PEG)i-Se core was synthesized by the reduction of commercially available polyethylene glycol) sorbitol hexaoleate (Sorb(PEG)i-Oe) core using of Pd / C hydrogenation followed by the standard procedure. An oven dried 50 ml round bottom flask was charged with purified Sorb(PEG)i-Oe (5.0 g, 2.5 mmol, 1.0 equiv), Ethanol (6.0 ml, 0.2 M) and Pd / C (1 .0 g, 20 mol%). The flask was equipped with a stir bar and the resulting suspension was sealed with a septum under an atmosphere of hydrogen gas supplied with a balloon. The reaction mixture was stirred vigorously at room temperature for 24 h and monitored by TLC. After the full consumption of the starting material, the suspension was filtered two times until a clear solution was obtained through a pad of celite bed. The crude mixture was concentrated in vacuo and passed through a pad silica gel to afford Sorb(PEG)i-Se as a white solid in 40% isolated yield (2.1 g, 1.0 mmol, unoptimized reaction condition).

[0147] Spectral data for Sorb(PEG)i-S6:1H NMR (500 MHz, CDCI3) 5 0.78 - 0.99 (m, 18H), 1.26 (s, 170H), 1.50 - 1.74 (m, 12H), 2.32 (ddd, J= 7.9, 5.5, 2.4 Hz, 12H), 3.47 - 3.90 (m, 133H), 4.23 (p, J= 3.0 Hz, 12H).13C NMR (126 MHz, CDCI3) 5 14.22, 22.79, 25.01 , 29.24, 29.38, 29.46, 29.58, 29.71 , 29.75, 29.78, 29.79, 32.02, 34.31 , 63.44, 69.31 , 70.63, 70.65, 70.67, 70.72, 76.91 , 77.16, 77.41 , 173.93.

[0148] Synthesis of tetrakis(4-oleyl benzoate) methane core (O4 core)

[0149] The compound was synthesized following a reported procedure. In a 100 ml flame- dried round bottom flask equipped with a stir bar, 4,4',4",4"'-Methanetetrayltetrabenzoic acid (0.5 g, 1 .0 mmol, 1 .0 equiv) and dichloromethane (DCM) (10.0 ml, 0.1 M) were combined under a nitrogen atmosphere applied by a balloon on top of the round bottom flask. The resulting mixture was placed in an ice bath for 10 minutes. One drop of DMF was added as the catalyst, followed by the addition of oxalyl chloride (0.9 ml, 10.0 mmol, 10.0 equiv). Asecond needle was placed on top of the round bottom flask to allow for the extrusion of generated gas during the reaction. After gas evolution ceased, the ice bath was removed, and the reaction mixture was allowed to warm up to room temperature and stirred at ambient temperature for 3 hours. The volatile components were then removed under reduced pressure, and the resulting yellow solid was redissolved in DCM. The reaction mixture was placed in an ice bath and cooled down to 0 °C, followed by the addition of oleyl alcohol (3.2 ml, 10.0 mmol, 10.0 equiv) under a nitrogen atmosphere applied by a balloon on top of the round bottom flask. The resulting reaction mixture was stirred at room temperature for 24 hours. The solvent was removed under reduced pressure, and the compound was further purified using column chromatography (30 mm X 250 mm, hexane: ethyl acetate 10:1 , 3:1 , 2:1 , and 1 :1). This process afforded the desired compound as a yellow oil in 33% yield (0.33 mmol, 0.50 g).

[0150] Spectral data for 04-core:1H NMR (500 MHz, CDCI3) 5 0.78 - 0.81 (m, 12H), 1.10 - 1.30 (m, 106H), 1.66 (p, J = 6.8 Hz, 8H), 1.88 - 1.98 (m, 16H), 4.21 (t, J = 6.6 Hz, 8H), 5.23 - 5.29 (m, 8H), 7.19 - 7.25 (m, 8H), 7.83 - 7.91 (m, 8H).13C NMR (126 MHz, CDCI3) 5 14.22, 22.80, 26.14, 27.29, 27.32, 28.85, 29.29, 29.32, 29.37, 29.43, 29.44, 29.54, 29.64, 29.86, 29.88, 32.02, 65.29, 65.31 , 76.91 , 77.16, 77.41 , 129.02, 129.39, 129.85, 130.08, 130.09, 130.82, 150.10, 166.21.

[0151] Synthesis of tetrakis(4-stearyl benzoate) methane core (S4core)

[0152] In a 100 ml round bottom flask, 4,4',4",4"'-methanetetrayltetrabenzoic acid (0.5 g, 1 .0 mmol, 1 .0 equiv) was combined with n-propanol (30.00 ml, 0.03 M), and one drop of H2SO4 was added as the catalyst. The reaction mixture was refluxed at 100 °C for 24 hours. After cooling down to room temperature, propanol was removed under reduced pressure, and the resulting solid mixture was dissolved in DCM. The solution was washed with a saturated solution of K2CO3 and dried over Na2SO4. The solvent was then removed under reduced pressure, and the desired product was obtained as a white solid, which was used in the next step without further purification.

[0153] In a 100 ml round bottom flask, 4,4',4",4"'-methanetetrayltetra(n-propyl benzoate) (0.13 g, 0.2 mmol, 1 .0 equiv) and stearyl alcohol (4.1 g, 40.0 mmol, 200 equiv) were combined, followed by the addition of one drop of H2SO4 as the catalyst. The reaction mixture was placed under high vacuum and then placed in an oil bath with the temperature set at 120 °C. The progress of the reaction was monitored using TLC, and after 24 hours, TLC indicated the full consumption of the starting materials. The oil bath was removed, and the reaction mixture was allowed to cool down to room temperature. The reaction mixture was dissolved in DCM, washed with K2CO3, and dried over Na2SO4. The solvent was thenremoved under reduced pressure, and the resulting crude mixture was purified via column chromatography (30 mm X 250 mm) using hexane: DCM (1 :1 ) and then hexane: ethyl acetate (1 :2) as the eluent. This process afforded the desired compound as a white solid in 83% yield (0.17 mmol, 0.25 g).

[0154] Spectral data for Au-lipid complex: 1 H NMR (500 MHz, CDCI3) 5 0.88 (t, J = 6.9 Hz, 12H), 1 .25 (s, 119H), 1 .37 - 1 .46 (m, 9H), 1 .73 (p, J = 6.8 Hz, 8H), 4.29 (t, J = 6.7 Hz, 8H), 7.27 - 7.32 (m, 8H), 7.91 - 7.97 (m, 8H). 13C NMR (126 MHz, CDCI3) 5 14.26, 22.84, 26.18, 28.87, 29.43, 29.51 , 29.68, 29.74, 29.79, 29.81 , 29.82, 29.83, 29.84, 32.07, 65.37, 76.91 , 77.16, 77.41 , 129.05, 129.41 , 130.85, 150.13, 166.30.

[0155] Synthesis of tetraoleyl pentaerythritol core (Pe-O4 core)

[0156] The compound was synthesized following a reported procedure. In a 100 ml flame- dried round bottom flask equipped with a stir bar, oleic acid (3.2 ml, 10.0 mmol, 10.0 equiv) and DCM (10.0 ml, 0.1 M) were combined under a nitrogen atmosphere applied by a balloon on top of the round bottom flask. The resulting mixture was placed in an ice bath for 10 minutes. One drop of DMF was added as the catalyst, followed by the addition of oxalyl chloride (1 .8 ml, 20.0 mmol, 20.0 equiv). A second needle was placed on top of the round bottom flask to allow for the extrusion of generated gas during the reaction. After gas evolution ceased, the ice bath was removed, and the reaction mixture was allowed to warm up to room temperature and stirred at ambient temperature for 3 hours. The volatile components were then removed under reduced pressure, and the resulting compound was redissolved in DCM. The reaction mixture was placed in an ice bath and cooled down to 0 °C, followed by the addition of oleyl pentaerythritol (0.14 g, 1 .00 mmol, 1 .00 equiv) and pyridine (0.8 ml, 10.0 mmol, 10.0 equiv) under a nitrogen atmosphere applied by a balloon on top of the round bottom flask. The resulting reaction mixture was stirred at room temperature for 24 hours. The crude mixture of the reaction was washed with NaOH (0.1 M, 3x10 ml) followed by washing with HCI (0.1 M, 1x10 ml). The organic layer was dried over Na2SC>4 and the solvent was removed under reduced pressure. The compound was further purified using column chromatography (30 mm X 250 mm, hexane: DCM 10:1 , 3:1 , 2:1 , and 1 :1). This process afforded the desired compound as a yellow oil in 34% yield (0.34 mmol, 0.40 g).

[0157] Spectral data for Au-lipid complex:1H NMR (500 MHz, CDC ) 5 0.89 (d, J= 6.7 Hz, 12H), 1.28 (dt, J = 14.9, 4.8 Hz, 91 H), 1.51 - 1.66 (m, 13H), 2.01 (q, J= 6.6 Hz, 16H), 2.30 (t, J = 7.6 Hz, 8H), 4.11 (s, 8H), 5.27 - 5.44 (m, 8H).13C NMR (126 MHz, CDCI3) 5 14.26, 22.83, 25.00, 27.33, 27.37, 29.27, 29.28, 29.33, 29.47, 29.48, 29.68, 29.86, 29.92, 32.08, 34.25, 41.97, 62.28, 76.91 , 77.16, 77.41 , 129.86, 130.16, 173.37.

[0158] Synthesis of oc-HDL NPs. Prior to each synthesis of oc-HDL NPs, fresh solutions of each reagent were prepared,[oc-HDL NP Size ± SD (nm) Zeta ± SD (mV) ] Aqueous solutions of Apo- Al (1 .4|PE-S4[l5.00 ± 4.93 1-14.73 ± 1.02 | mg / mL) and [PE-O4 _ [12.51 ± 3.75 [18.88 ± 0.43 ] dichloromethane [sorb(PEG)n-S6l 8.14 ± 9.391 ]-13.23 ± 0.35 ] (DCM) solutions of the | organic scaffold and the phospholipid DPPC (1 mM)were prepared in sufficient volumes for the required syntheses. To initiate particle assembly, the nonscaffold nanoparticle components are added sequentially to the core scaffolds in 6 mL glass vials. In a typical assembly, Apo-A1 (2 nmol from a 1 .4 mg / mL suspension in PBS) was combined with the lipid conjugate core scaffold (1 nmol in DCM), followed by the addition of DPPC (200 nmol in DCM). This combined suspension is then diluted in PBS to a final core concentration of 10 pM in PBS. Next, the suspension was subjected to six rounds of alternating bath sonication (45 s) and vortexing (45s). The suspension underwent stirring using a magnetic stir bar for 2 hours with the cap of the glass vial removed to allow evaporation of the organic solvent (DCM). The particles were transferred to Eppendorf tubes and centrifuged by desktop centrifugation at 10,000 g for 10 minutes to remove aggregates, if any. The particles were then filtered using spin columns (MilliPore; 0.5 mL 50 kDa MWCO). The column was first rinsed in PBS for 10 min at 10,000 g at room temperature, prior to three rounds of spin filtration of the particles under the same conditions. An aliquot of PBS (500 pL) was added to the column after each spin. After the final round of filtration, the small volume of concentrated particles 20 pL) was diluted to 100-200 pL in PBS. Particle protein concentration was determined by bicinchoninic acid assay (BCA), with spectroscopic measurement of absorbance at 562 nm after incubation with BCA reagents at 37 °C for 30 min. For all downstream assays, the concentration of oc-HDL NPs was determined by BCA quantification of protein; and protein concentration was held constant across all groups.

[0159] Dynamic light scattering (DLS), zeta potential, and transmission electron microscopy (TEM) measurements reveal all cores form small (12-20 nm), negatively charged, spherical structures that mimic native spherical HDL. Size and zeta potentials of oc-HDL nanoparticles made with different organic cores, and with different phospholipid tails are set out in the tables below.Table 1Table 2

[0160] To determine the number of Apo-A1 molecules per particle, particles were crosslinked with bis(sulfosuccinidimidyl)suberate (BS3) (Sigma Aldrich), which stabilizes any higher order oligomerization states of Apo-Al in oc-HDL NPs upon assembly. All oc-HDL NPs contained multiple Apo-A1 molecules (monomers, dimers, trimers, tetramers, pentamers) consistent with native spherical HDL (Figure 5).

[0161] oc-HDL NPs Transport Cholesterol from J774 macrophages to HepG2 hepatocytes. J774 macrophages labeled with fluorescently labeled cholesterol (Top-Fluor Cholesterol) were treated with 256 nM (Apo-A1 concentration) solutions of nanoparticles or controls. After 6 hours, efflux media was transferred to HepG2 and was incubated for 10 hours. The PBS control represented efflux / influx due to diffusion alone. No significant differences in cholesterol efflux or influx were detected across Apo-A1 and oc-HDL NPs (one-way ANOVA) (Figure 6). These data demonstrate that all oc-HDL NPs are capable of transporting free cholesterol present on the cellular membrane from macrophages to hepatocytes, resembling the reverse cholesterol transport function of native HDL.

[0162] oc-HDL NPs Load Cholesteryl Esters. The capacity of oc-HDL NPs to load cholesteryl esters was assessed by synthesizing oc-HDL NPs with cholesteryl lineolate, oneof the most abundant cholesteryl esters present in native HDL. Particles were purified by spin filtration to remove free cholesteryl lineolate and quantified by BCA and DLS. The difference in size of particles with and without cholesterol is set out in Table 3.Table 3 |

[0163] The amount of cholesteryl was measured using the Amplex Red Cholesterol Assay Kit. Total cholesterol relative to Apo-A1 protein was plotted. No significant differences in cholesterol content was found among all particles (one-way ANOVA) (Figure 7).

[0164] oc-HDL NPs reduce inflammation in human macrophage cells. THP1 - Dual macrophage cells were treated with LPS to induce NF-KB activity assayed by a colorimetric QUANTIBIue assay as a read out of inflammation. Treatment with oc-HDL NPs significantly inhibited LPS-induced NF-KB activity more than free Apo-A1 (one-way ANOVE) (*p < 0.05, **p < 0.01 , ***p < 0.001 ) (Figure 8).

[0165] These results show that the synthetic oc-HDL-NP described herein have functions similar to natural HDL particles and are useful as therapeutics to treat disease or disorders associated with cholesterol transport and immune response.EXAMPLE 3-CHARACTERIZATION OF OC-CORE PARTICLES

[0166] Additional oc-core particles were developed and assessed for cholesterol transport efficiency.

[0167] Table 4 shows the properties of new HDL core particles as measured by dynamic light scattering (DLS).Table 4

[0168] DLS and TEM analysis (Figure 9) reveal oc-HDL NPs are small, negatively- charged, spherical nanostructures that mimic native HDL. In vitro cholesterol efflux assays show that oc-HDL NPs efficiently transfer free cholesterol from macrophages and upregulate ABCA1 expression, supporting their function in reverse cholesterol transport (Figure 10). Statistically significant differences are denoted between oc-HDL NPs and apo-A1 (x) and oc- HDL NPs and HDL2 (w) in +CAMP conditions and between oc-HDL NPs and apo-A1 (y) and oc-HDL NPs and HDL2 (z) in -cAMP conditions.

[0169] Native HDLs possess anti-inflammatory function and have been demonstrated to reduce NF-KB and JNK signaling. Oc-HDLs were tested for their ability to inhibit inflammatory signals. LPS-induced NF-KB and p-JNK signaling was reduced after cotreatment with DOPS oc-HDL NPs in an NF-KB reporter line (THP-1 cells) (Figure 11).

[0170] Cholesteryl lineolate, a predominant cholesteryl ester found in native HDL, was incorporated into the nanoparticles during the synthesis of oc-HDL NPs as a proxy to assess cholesterol loading / hydrophobic drug loading of the nanoparticles. Particles containing DOPS, a negatively charged phospholipid with unsaturated lipid tails, load more cholesterol lineolate than any other construct (Figure 12).

[0171] The results above demonstrate that the ability to synthesize HDL NP mimetics using multiple organic cores (oc) and phospholipids (PL) leads to particles with similar physical properties (i.e., size, shape) and negative surface charge. All the oc-HDL NPs support cholesterol efflux. Additionally, oc-HDL NPs comprising DOPC and DOPS demonstrate enhanced efflux through ABCA1 -independent mechanisms. It was shown that DOPS PE-O4 possessed the greatest suppression of LPS-induced inflammation and the highest cholesteryl ester loading, making it a promising candidate for further studies.

[0172] Numerous modifications and variations to the disclosure, as set forth in the embodiments and illustrative examples described herein, are expected to occur to thoseskilled in the art. Consequently only such limitations as appear in the appended claims should be placed on the disclosure.

Claims

What is claimed is:1 . A spherical lipoprotein-like nanoparticle (HDL-NP) comprising: a core and a shell surrounding and attached to the core, wherein the core comprises a lipid conjugated organic scaffold comprising a dendritic lipid scaffold.

2. The spherical HDL-NP of claim 1 , wherein the dendritic lipid scaffold comprises pentaerythritol distearate (PE-S2), pentaerythritol dioleate (PE-O2), pentaerythritol tetrastearate (PE-S4), pentaerythritol tetraoleate (PE-O4), sorbitol(polyethylene glycol)n-hexasterate (Sorb(PEG)n-Se) sorbitol(polyethylene glycol)n- hexaoleate (Sorb(PEG)n-Oe), tetraoleate (O4) or tetrastearate (S4).

3. The spherical HDL-NP of claim 1 or 2, wherein the shell is a lipid shell.

4. The spherical HDL-NP of any one of claims 1 -3, wherein the shell is a lipid bilayer or monolayer.

5. The spherical HDL-NP of any one of claims 1 -4, wherein the spherical HDL-NP has a surface zeta potential of about -5 to about -60 millivolts (mV).

6. The spherical HDL-NP of any one of claims 1 - 5, wherein the organic core comprises a headgroup-modified fatty acid.

7. The spherical HDL-NP of claim 6, wherein the headgroup-modified fatty acid comprises C12, C14, C16, C18 or C20 tails.

8. The method of claim 7, wherein the fatty acid is monounsaturated or polyunsaturated.

9. The spherical HDL-NP of any one of claims 6-8, wherein the fatty acid is selected from the group consisting of lauric acid, myristic acid, palmitic acid, stearic acid, arachidic acid, oleic acid, linoleic acid, linolenic acid, eleostearic acid, and arachidonic acid.

10. The spherical HDL-NP of any one of claims 1 -9, wherein the shell comprises components selected from the group consisting of 1 ,2-dipalmitoyl-sn-glycero-3- phosphocholine (DPPC), 1 ,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC), 1 ,2-diolinoleoyl-sn-glycero-3-phosphocholine (DLPC), 1 ,2-dioleoyl-sn-glycero-3-phospho-L-serine (DOPS), 1 ,2-distearoyl-sn-glycero-3-phospho-(1'-rac-glycerol) (18:0 PG), or 1 ,2-dilinoleoyl-sn-glycero- 3-phospho-(1 '-rac-glycerol) (18:2 PG).11 . The spherical HDL-NP of any one of claims 1-10, wherein the HDL nanoparticle comprises DPPC Polyethylene glycol) sorbitol hexaoleate (Sorb(PEG)l-Oe), DPPC Polyethylene glycol) sorbitol hexastearate (Sorb(PEG)l -Se), DPPC PE-S4, DPPC PE-O4, DOPC PE-O4 or DOPS PE-O4.

12. The spherical HDL-NP of any one of claims 1 -11 , wherein a small molecule with a plurality of terminal functional groups is coupled to one or more lipid to form the dendritic lipid scaffold.

13. The spherical HDL-NP of claim 12, wherein the small molecule pentaerythritol, sorbitol and tetraA7s(4-functionalized phenyl)methane.

14. The spherical HDL-NP of claim 12 or 13, wherein the plurality of functional groups are hydroxyls.

15. The spherical HDL-NP of claim 12 or 13, wherein the plurality of functional groups are amino groups.

16. The spherical HDL-NP of any one of claims 12-15, wherein the plurality of functional groups are carboxylates.

17. The spherical HDL-NP of any one of claims 1 -16, wherein the spherical HDL-NP has a diameter of about 5-60 nm, 10-50 nm, 5-40 nm, 10-30 nm, 10-20 nm, 5-30 nm, 5-20 nm, 5- 15 nm, 5-10 nm, 8-15 nm, or 8-12 nm.

18. The spherical HDL-NP of any one of claims 1 -17, wherein the spherical HDL-NP has a negative zeta potential similar to or more negative than human HDL.

19. The spherical HDL-NP of claim 18, wherein the zeta potential is -5 mV to -25 mV.

20. The spherical HCL-NP of any one of claims 1 -19, wherein the HDL-NP is capable of associating with cholesterol21 . The spherical HDL-NP of any one of claims 1-20, wherein the spherical HDL-NP has a more efficient cholesterol transport capacity than that of apolipoprotein or a synthetic HDL nanoparticle with a gold core.

22. The spherical HDL-NP of any one of claims 1 -21 , further comprising apolipoprotein.

23. The spherical HDL-NP of claim 22, wherein the apolipoprotein is an apolipoprotein A, apolipoprotein B, apolipoprotein C, apolipoprotein D, apolipoprotein E or apolipoprotein H.

24. The spherical HDL-NP of claim 21 or 22 wherein the apolipoprotein is selected from the group consisting of apo A-l, apo A-ll, apo A-IV, apo A-V, apo B48, apo B100, apo C-l, apo C-ll, apo C-lll, apo C-IV, apolipoprotein D, apolipoprotein E and apolipoprotein H.

25. The spherical HDL-NP of any one of claims 22-24, further comprising apolipoprotein A-l (ApoA1).

26. The spherical HDL-NP of any one of claims 1-25, wherein the HDL-NP has a hydrodynamic diameter of greater than 8 nm, or about 8 nm to about 30 nm.

27. The spherical HDL-NP of any one of claims 1-26, wherein the HDL-NP has a hydrodynamic diameter of 8 nm-17nm.

28. The spherical HDL-NP of any one of claims 1-26, wherein the HDL-NP has a hydrodynamic diameter of 10 nm-15 nm.

29. The spherical HDL-NP of any one of claims 1-26, wherein the HDL-NP has a hydrodynamic diameter of 12 nm-14 nm.

30. The spherical HDL-NP of any one of claims 1-26, wherein the HDL-NP organic core comprises PE-S4 and has a diameter of about 10 nm to about 20nm and a zeta potential between about -10 mV to about -20 mV.31 . The spherical HDL-NP of any one of claims 1-26, wherein the HDL-NP organic core comprises PE-O4 and has a diameter of about 8 nm to about 17 nm and a zeta potential between about -5 mV to about -15 mV.

32. The spherical HDL-NP of any one of claims 1-26, wherein the HDL-NP organic core comprises Sorb(PEG)n-Se and has a diameter of about 10 nm to about 30 nm and a zeta potential between about -10 mV to about -20 mV.

33. The spherical HDL-NP of any one of claims 1 -26, wherein the HDL-NP organic core comprises Sorb(PEG)n-Oe and has a diameter of about 10 nm to about 25 nm and a zeta potential between about -15 mV to about -25 mV.

34. The spherical HDL-NP of any one of claims 1 -26, wherein the HDL-NP organic core comprises S4 and has a diameter of about 7 nm to about 25 nm and a zeta potential between about -10 mV to about -20 mV.

35. The spherical HDL-NP of any one of claims 1-26, wherein the HDL-NP organic core comprises O4 and has a diameter of about 7 nm to about 25 nm and a zeta potential between about -10 mV to about -20 mV.

36. The spherical HDL-NP of any one of claims 1-35, further comprising a therapeutic agent or natural lipophilic molecule adsorbed or linked to the HDL-NP.

37. The spherical HDL-NP of claim 36, wherein the therapeutic agent is a therapeutic nucleic acid, an anti-cancer agent, a chemotherapeutic agent, an antioxidant or an antiinflammatory agent.

38. A pharmaceutical composition comprising the spherical HDL-NP of any one of claims 1-37 and a pharmaceutically acceptable excipient, diluent or carrier.

39. A method for treating a cancer, the method comprising: administering to a subject having a cancer a spherical lipoprotein-like nanoparticle (HDL-NP) comprising: a core and a shell surrounding and attached to the core, wherein the core comprises a lipid conjugated organic scaffold in an effective amount to treat the cancer, wherein the lipid conjugated organic scaffold comprises a dendritic lipid scaffold.

40. A method of treating an inflammatory disorder comprising, administering to a subject having an inflammatory disorder a spherical lipoprotein-like nanoparticle (HDL-NP) comprising: a core and a shell surrounding and attached to the core, wherein the core comprises a lipid conjugated organic scaffold in an effective amount to treatthe inflammatory disorder, wherein the lipid conjugated organic scaffold comprises a dendritic lipid scaffold.41 . The method of claim 39 or 40, wherein the lipid conjugated organic scaffold comprises pentaerythritol distearate (PE-S2), pentaerythritol dioleate (PE-O2), pentaerythritol tetrastearate (PE-S4), pentaerythritol tetraoleate (PE-O4), sorbitol(polyethylene glycol)n-S6 (Sorb(PEG)n-Se) or sorbitol(polyethylene glycol)n-06 (Sorb(PEG)n-Oe), tetrastearate (S4) or tetraoleate (O4).

42. The method of any one of claims 39 to 41 , wherein the spherical HDL-NP is a spherical HDL-NP of any one of claims 1-37 or pharmaceutical composition of claim 38.

43. The method of any one of claims 39 to 42, wherein the subject is a mammal.

44. The method of any one of claims 39 to 43, wherein the subject is a human.

45. The method of any one of claims 39 to 44, wherein the spherical HDL-NP is administered intravenously, subcutaneously, intramuscularly, topical or orally.

46. A method of reducing NF-kB and / or TNF-a activity, comprising administering to a subject an effective amount of a composition comprising the spherical HDL-NP of any one of claims 1-37 or pharmaceutical composition of claim 38.

47. A method for making a spherical lipoprotein-like nanoparticle (HDL-NP) comprising: -preparing an organic scaffold solution;-contacting the organic scaffold solution with apolipoprotein to generate an organic scaffold-Apo solution;-contacting the organic scaffold-Apo solution with a lipid solution to generate an organic scaffold-Apo-lipid solution;-admixing the organic scaffold-Apo-lipid solution by vortexing and / or sonication to form HDL-NP;-isolating the HDL-NP.

48. The method of claim 47, wherein the HDL-NP comprises a core and a shell surrounding and attached to the core, wherein the core comprises a lipid conjugated organic scaffold.

49. The method of claim 47 or 48, wherein the organic scaffold solution comprises any one of pentaerythritol distearate (PE-S2), pentaerythritol dioleate (PE-O2), pentaerythritol tetrastearate (PE-S4), pentaerythritol tetraoleate (PE-O4), sorbitol(polyethylene glycol)n-S6 (Sorb(PEG)n-Se) or sorbitol(polyethylene glycol)n-06 (Sorb(PEG)n-Oe) tetrastearate (S4) or tetraoleate (O4).

50. The method of any one of claims 47 to 49, wherein the organic scaffold comprises a headgroup-modified fatty acid.51 . The method of claim 50, wherein the headgroup-modified fatty acid comprises C12, C14, C16, C18 or C20 tails.

52. The method of claim 50 or 51 , wherein the fatty acid is monounsaturated or polyunsaturated.

53. The method of any one of claims 47 to 52, wherein the lipid components of the organic scaffold are selected from the group consisting of lauric acid, myristic acid, palmitic acid, stearic acid, arachidic acid, oleic acid, linoleic acid, linolenic acid, eleostearic acid, and arachidonic acid.

54. The method of any one of claims 47 to 53, wherein the apolipoprotein is apolipoprotein A-l.

55. The method of any one of claims 48 to 54, wherein the shell comprises components selected from the group consisting of 1 ,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC), 1 ,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC), 1 ,2-diolinoleoyl-sn-glycero-3- phosphocholine (DLPC), 1 ,2-dioleoyl-sn-glycero-3-phospho-L-serine (DOPS), 1 ,2-distearoyl- sn-glycero-3-phospho-(1'-rac-glycerol) (18:0 PG), or 1 ,2-dilinoleoyl-sn-glycero-3-phospho- (1 '-rac-glycerol) (18:2 PG).

56. The method of any one of claims 40 to 55, wherein the HDL nanoparticle comprises DPPC Polyethylene glycol) sorbitol hexaoleate (Sorb(PEG)l-Oe), DPPC Polyethylene glycol) sorbitol hexastearate (Sorb(PEG)l-Se), DPPC PE-S4, DPPC PE-O4, DOPC PE-O4 or DOPS PE-O4.

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