NAD(h) nanoparticles and methods of reducing vein graft injury and improving vein graft health

Abstract

The present technology provides methods of reducing vein graft injury comprising administering an effective amount of a bioavailable nanoparticle comprising NAD+ and / or NADH to the vein to be grafted prior to surgical implantation of the vein graft in a subject. Additionally, the present technology provides methods of improving vein graft health and preventing or treating post-operative restenosis and also provides bioavailable nanoparticles comprising NAD+ and / or NADH and a coating comprising a human cell membrane for use in such methods.

Description

NAD(H) NANOPARTICLES AND METHODS OF REDUCING VEIN GRAFT INJURY AND IMPROVING VEIN GRAFT HEALTHCROSS-REFERENCE TO RELATED APPLICATIONS

[0001] This application claims the benefit of and priority to U.S. Provisional Appl. No. 63 / 623,142, filed January 19, 2024, the contents of each of which are incorporated herein by reference in their entirety for any and all purposes.GOVERNMENT RIGHTS

[0002] This invention was made with government support under AI165977 awarded by the National Institutes of Health. The government has certain rights in the invention.FIELD

[0003] The present technology relates generally to the field of bioavailable nanoparticles comprising NAD(H) (i.e., NAD+and / or NADH) and use thereof in methods of reducing vein graft (VG) injury and improving VG health, e.g., via improving VG patency and survival as well as anti-restenotic therapy. The nanoparticles optionally include a human cell membrane such as a human platelet cell membrane.BACKGROUND

[0004] Autologous vein grafts (VG) are the most commonly used conduits for coronary and peripheral artery bypass procedures but are prone to maladaptive remodeling that leads to progressive luminal loss and VG failure. Despite advances in surgical techniques and pharmacotherapies, the rate of VG failure remains high at approximately 15% at 1 year and up to 50% at 10 years. VG failure can significantly impact patient quality of life and increase the risk of morbidity and mortality. Repeat revascularization procedures are invasive and have limited efficacy. The development of new, effective paradigms to prevent VG failure remains a major challenge.

[0005] The principal mechanism of VG failure is intimal hyperplasia (IH), featuring progressive accumulation of intimal cells and luminal loss. Although its mechanism is not completely understood, prior studies suggest a critical role of endothelial cell (EC)dysfunction and other pathophysiologies like thrombosis that collectively lead to VG failure. EC dysfunction can result from acute pre-bypass damage (e.g., improper storage). During the intraoperative stage, dysfunctional ECs primarily feature an acute degenerative phenotype, characterized by loss of viability and integrity and activation of thrombogenic and inflammatory responses. Despite decades of progress in organ preservation for transplantations, non-buffered saline and autologous blood remain the primary option for intraoperative storage of autologous VG, which are widely known to damage ECs and contribute to VG failure.

[0006] Similarly, the long-term patency of reconstructed blood vessels (e.g., via endovascular interventions such as balloon angioplasty and stenting) has been consistently compromised, primarily caused by restenosis - the re-narrowing or re-occlusion of the target vessels. Over the past two decades, several generations of drug-eluting stents and drug-coated balloons have been introduced to specifically address the persistent recurrence of restenosis. Through device-enabled intraluminal delivery of often toxic drugs (e.g., paclitaxel and sirolimus), anti-restenotic effects could be achieved via inhibiting the excessive proliferation of vascular smooth muscle cell (SMC), or intimal hyperplasia (IH). In comparison to bare metal stents and plain balloon angioplasty, these drug-eluting / -coated devices are associated with reduced restenotic rates and hence improved post-intervention outcomes.

[0007] Emerging evidence suggests that drug-eluting / -coated devices create excessive damage to fragile endothelial cells (EC), leading to impaired re-endothelialization and hence increased risk of thrombosis and related adverse events. The use of dual-antiplatelet therapy following endovascular interventions is required to reduce the thrombotic risk, yet its extended use not only increases risk of bleeding but also incurs significant financial burdens. Indeed, while the anti-restenotic drugs successfully block SMC proliferation and IH, they simultaneously exacerbate the apoptotic and pro-inflammatory responses of the adjacent ECs. A growing body of literature confirms that arterial / aortic ECs are far more sensitive to the cytotoxicity incurred by commonly used anti-restenotic agents than SMCs. Hence, a therapeutic regime that protects ECs but inhibits SMC proliferation would be desirable.SUMMARY

[0008] The present technology provides bioavailable nanoparticles (NPs) that include the oxidized and / or reduced forms of nicotinamide dinucleotide, z.e., NAD+and NADH respectively, or NAD(H) for short. It has been discovered that the present NPs and methods advantageously selectively block the excessive proliferation of SMC and IH, while preserving or even rescuing the integrity of EC following endovascular interventions, including for vein grafts and angioplasty.

[0009] In one aspect, the present technology provides methods of reducing vein graft injury comprising administering an effective amount of a bioavailable nanoparticle comprising NAD+and / or NADH to the vein to be grafted prior to surgical implantation of the vein graft in a subject. The effective amount of the bioavailable nanoparticle may be administered by storing the vein to be grafted in an aqueous solution comprising the effective amount of the bioavailable nanoparticle.

[0010] In another aspect, the present technology provides methods of improving vein graft health and bioavailable nanoparticles for same. The methods include administering an effective amount of a bioavailable nanoparticle comprising NAD+and / or NADH and a coating comprising a human cell membrane to a subject post-surgical implantation of the vein graft in the subject, wherein the human cell membrane is selected from the group consisting of a human platelet membrane, a human red blood cell membrane, a human white blood cell membrane, mesenchymal stem cell membrane, and a human macrophage cell membrane.

[0011] In still another aspect, the present technology provides methods of preventing or treating post-operative restenosis and bioavailable nanoparticles for same. The methods include administering an effective amount of a bioavailable nanoparticle comprising NAD+and / or NADH and a coating comprising a human cell membrane to a subject after surgical reconstruction of a blood vessel in the subject, wherein the human cell membrane is selected from the group consisting of a human platelet membrane, a human red blood cell membrane, a human white blood cell membrane, mesenchymal stem cell membrane, and ahuman macrophage cell membrane. In some embodiments, the surgical reconstruction of the blood vessel in the subject comprises balloon angioplasty and / or placement of a stent.

[0012] In one aspect, bioavailable nanoparticles for use in the present methods include NAD+and / or NADH and a coating comprising a human cell membrane. The human cell membrane may be selected from the group consisting of a human platelet membrane, a human red blood cell membrane, a human white blood cell membrane, mesenchymal stem cell membrane, and a human macrophage cell membrane. The bioavailable nanoparticles may include an inorganic core and NAD+and / or NADH, and a coating including a lipid bilayer. The inorganic core may be calcium phosphate or a metal organic framework (MOF). In another aspect, the bioavailable nanoparticle may be a nanoparticle comprising a disulfide-containing lipopeptide as disclosed in USSN 63 / 443,675 (filed 2 / 6 / 2023 and titled “Disulfide-containing lipopeptides, nanoparticles and methods of use”), incorporated by reference herein and for all purposes. In still another aspect, the bioavailable nanoparticle may be a nanoparticle comprising a poly(amidoamine) oligomer, as disclosed in USSN 63 / 546,129 (filed 10 / 27 / 2023 and titled “Lipid nanoparticls formed by lipidoids for efficient delivery of nucleotide drugs and biologies”), incorporated by reference herein and for all purposes.BRIEF DESCRIPTION OF THE DRAWINGS

[0013] FIGS. 1A-1F show characterization data for an illustrative embodiment of a biomimetic platelet membrane-coated and NAD+-loaded CaP-NP (P-NAD+-CaP NP). FIG. 1A shows hydrodynamic diameters of platelet vesicle, NAD+-NP and P-NAD+-NP characterized by DLS. FIGS. IB and 1C show morphology of NAD+-NP and P-NAD+-NP respectively, characterized by transmission electron microscopy. FIG. ID shows Zeta potentials of platelet derived vesicles and NAD+-NP with different platelet membrane to NAD+-NP weight ratios were measured. Data are presented as mean ± s.d. (n=3). Statistical significance was calculated by one-way analysis of variance (ANOVA) with Tukey’s post hoc test, n.s., no significance (i.e., P > 0.05). FIG. IE shows zeta potentials of platelet vesicles, non-biomimetic NAD+-NP, and the biomimetic P-NAD+-NP measured by dynamic light scattering. FIG. IF shows hydrodynamic diameters of P-NAD+-NP stored at 4°C for one week by DLS.

[0014] FIGS. 2A-2E show that ex vivo VG storage in non-buffered (saline) and buffered (Plasma-Lyte) solutions led to rapis NAD+depletion and EC dysfunction. FIG. 2A shows that storage in saline or Plasma-Lyte (30 min) reduced NAD+in human saphenous VG (n=4). lOpM NAD+-NP reduced the reduced the mRNA expression of dysfunctional EC (EC degeneration) markers P-Selectin (FIG. 2B) and inflammasome genes, NLRP3 (FIG. 2C) and ILlb (FIG. 2D), and rescued the expression of EC function marker eNOS after 6hr ex vivo exposure (FIG. 2E). Mean+SEM. Paired One-Way ANOVA followed by Tukey test. *p<0.05.

[0015] FIGS. 3A-3B. Pre-bypass, ex vivo NAD+-replenishing therapy mitigates IH after rat VG bypass procedure in vivo. FIG. 3A is a schematic illustration of the pre-bypass VG storage and subsequent bypass procedure in male SD rats. FIG. 3B shows representative ultrasound imaging of VG 2 months post-bypass, following 3hr intraoperative storage in saline with or without lOpM NAD+-NP at room temperature. FIG. 3C shows quantification of lumen diameter and distal wall thickness. Mean+SEM. n=4-7. Unpaired Student’s t-test. *p<0.05.

[0016] FIGS. 4A-4B show an illustrative embodiment of present technology in which Platelet membrane-coated, NAD+-loaded CaP-NP (P-NAD+-NP) inhibits SMC proliferation while differentially preserving EC health. Primary culture of human aortic SMC (HASMC) and human aortic EC (HAEC) were utilized to test the differential cellular outcomes of Ca- NP-enabled NAD+repletion. FIG. 4A shows viability and prolifereation results upon exposing 80-90% confluent and overnight starved HASMC to the respective treatments (equivalent to lOpM NAD+payload) with or without 20 ng / mL PDGF-BB to stimulate SMC proliferation, cell viability and proliferation were determined using bioluminescent CellTiter-Glo Cell Viability assay (72hr) and colorimetric BrdU Incorporation assay (24hr). Presented as Mean ± SEM (n=4). FIG. 4B shows viability and apoptosis results upon exposing fully confluent HAEC to the respective treatments (equivalent to lOpM NAD+payload) with or without 20 ng / mL TNFa to stimulate EC death, cell viability and apoptosis were determined using bioluminescent CellTiter-Glo Cell Viability assay (24hr) and Caspase3 / 7-Glo assay (4hr). Presented as Mean ± SEM (n=4). *P < 0.05, **P < 0.01, ***p < 0.001, determined by One-way ANOVA followed by post-hoc Tukey test.

[0017] FIGS. 5A-5C show an illustrative embodiment of the present technology in which a platelet membrane-coated CaP-NP demonstrated selective lesion-targetability and desirable biodistribution. FIG. 5A shows overall study design: ATTO550-tracrRNA-loaded biomimetic CaP-NP were intravenously injected immediately post procedure. The balloon- injured and non-injured carotid arteries were collected 6hr post procedure / inj ection for ex vivo imaging using a IVIS system (Ex / Em: 545 / 575 nm). FIG. 5B shows ex vivo fluorescence IVIS imaging of the injured (by balloon angioplasty) versus non-injured, contralateral carotid arteries. Quantitative analysis of the normalized fluorescence intensity was conducted. FIG. 5C shows ex vivo IVIS imaging of injured carotid artery versus major organs. L, H, A, S, L, K, Int, M, B represent liver, heart, aorta, spleen, lung, kidney, intestine (intes.), skeletal muscle (muse.), and brain. Quantitative analysis of the mean fluorescence intensity per unit mass in each organ or tissue shown in the ex vivo images.

[0018] FIGS. 6A-6D show an illustrative embodiment of present technology provides targeted NAD+delivery via the biomimetic P-NAD+-NP mitigated post-angioplasty IH in vivo. FIG. 6A summarizes study design in which the platelet membrane-coated NAD+-NP or the control groups were intravenously administered (one-time injection; 10 mg / kg NAD+payload or equivalent) immediately following carotid artery balloon angioplasty in male SD rats. FIG. 6B shows representative H&E histology images of IH lesions (low and high magnitudes) at day 14 post procedure. Neointima (N), media (M), and adventitia (Adv) are indicated, respectively. FIG. 6C shows morphometric analysis of IH based on intima-to-media (I / M) ratio. FIG. 6D shows results of qPCR analysis of mRNA expression changes in tissue homogenates of rat carotid arteries post angioplasty. Quantitation of mRNA level of SMC contractility / maturation gene (aSMA). n=4-8. Presented as Mean±SEM. *P<0.05. One-way ANOVA with Tukey post-hoc analysis.

[0019] FIGS. 7A-7C. show an illustrative embodiment of present technology in which targeted NAD+delivery via the biomimetic P-NAD+-NP accelerated re-endothelialization following angioplasty-induced EC denudation in vivo. FIG. 7A summarizes overall study design: Prototypic LP-CaP-NP was hybridized with platelet membrane coating to enable IH-lesion targetability as well as biocompatibility. The injectable NAD+-NP or the control groups were intravenously administered (one-time injection; 10 mg / kg NAD+payload or equivalent) immediately following carotid artery balloon angioplasty in male SD rats. Animals were injected with Evans Blue to visualize non-endothelium-covered luminal areas30min prior to euthanasia and macroscopic examination at day 6 post angioplasty. FIG. 7B shows representative images of Evans Blue-stained carotid arteries at day 6 post angioplasty and quantification of re-endothelialization (n=10-13). FIG. 7C shows results of qPCR analysis of mRNA expression changes in tissue homogenates of rat carotid arteries at day 6 post angioplasty. Quantitation of mRNA levels of EC signature gene representative of EC physiological function / integrity (eNOS), thrombogenicity (tissue factor / TF), impairment of EC healing (CXCL10), and inflammatory cytokine (ICAM1). Presented as Mean±SEM. *p<0.05. One-way ANOVA with Tukey post-hoc analysis.DETAILED DESCRIPTION

[0020] The following terms are used throughout as defined below. All other terms and phrases used herein have their ordinary meanings as one of skill in the art would understand.

[0021] As used herein and in the appended claims, singular articles such as “a” and “an” and “the” and similar referents in the context of describing the elements (especially in the context of the following claims) are to be construed to cover both the singular and the plural, unless otherwise indicated herein or clearly contradicted by context.

[0022] As used herein, “about” will be understood by persons of ordinary skill in the art and will vary to some extent depending upon the context in which it is used. If there are uses of the term which are not clear to persons of ordinary skill in the art, given the context in which it is used, “about” will mean up to plus or minus 10% of the particular term.

[0023] As used herein, the terms “effective amount” or “therapeutically effective amount,” or “pharmaceutically effective amount” refer to a quantity sufficient to achieve a desired therapeutic and / or prophylactic effect, e.g., an amount which results in the full or partial amelioration of disease, disorder, condition or symptoms of a subject (e.g., a human) in need thereof. For example, the effective amount may be an amount that reduces or prevents endothelial cell dysfunction, intimal hyperplasia, vein graft injury and / or restenosis, and / or promotes vein graft health. In the context of therapeutic or prophylactic applications, the amount of a composition administered to the subject will depend on thetype and severity of the disease and on the characteristics of the individual, such as general health, age, sex, body weight and tolerance to drugs. It will also depend on the degree, severity and type of disease. A person of ordinary skill in the art will be able to determine appropriate dosages depending on these and other factors. The compositions can also be administered in combination with one or more additional compounds. Multiple doses may be administered. Additionally or alternatively, multiple therapeutic compositions or compounds may administered. In the methods described herein, the compounds may be administered to a subject having one or more signs or symptoms of a disease or disorder described herein.

[0024] As used herein, the term “subject” refers to any animal that can experience vein graft injury, such as a mammal or a bird. In any embodiments, the mammal may be selected from primates, dogs, cats, rodents, horses, cattle, or pigs. In any embodiments, the subject (i.e., primate subject) is a human.

[0025] “Treating,” “treat,” “treated,” or “treatment” as used herein covers the treatment of a disease or disorder described herein (e.g., restenosis), in a subject, such as a human, and includes: (i) inhibiting or preventing a disease or disorder, i.e., arresting its development;(ii) relieving a disease or disorder, i.e., causing regression of the disorder; (iii) slowing progression of the disorder; and / or (iv) inhibiting, preventing, relieving, ameliorating, or slowing progression of one or more symptoms of the disease or disorder. Symptoms may be assessed by methods known in the art or described herein, for example, biopsy, histology, and blood tests to determine relevant enzyme levels, metabolites or circulating antigen or antibody (or other biomarkers), quality of life questionnaires, patient-reported symptom scores, and imaging tests.

[0026] “Ameliorate,” “ameliorating,” and the like, as used herein, refer to inhibiting, relieving, eliminating, or slowing progression of one or more symptoms.

[0027] “Metal organic framework” or “MOF” as used herein refers to the three- dimensional, porous, crystalline structure formed by metal ions and small organic ligands that coordinate to the metal ions. Thus, a “metal organic framework component” refers collectively to the individual component parts of the MOF, i.e., a metal ion and acoordinating ligand. For example, a zinc ion and 2-methylimidazole would be, respectively, the metal ion and coordinating ligand of the metal organic framework component for the MOF, zeolitic imidazolate framework-8, i.e., ZIF-8.

[0028] “Molecular weight” as used herein with respect to polymers refers to numberaverage molecular weights (Mn) and can be determined by techniques well known in the art including gel permeation chromatography (GPC). GPC analysis can be performed, for example, on a D6000M column calibrated with poly(methyl methacrylate) (PMMA) using triple detectors including a refractive index (RI) detector, a viscometer detector, and a light scattering detector, and TV, TV ’-dimethylformamide (DMF) as the eluent. “Molecular weight” in reference to small molecules and not polymers is actual molecular weight, not numberaverage molecular weight.

[0029] The terms “preventing” and “prophylaxis” as used herein refer to administering a pharmaceutical compound or medicament or a composition including the pharmaceutical compound or medicament to a subject before a disease, disorder, or condition fully manifests itself, to forestall the appearance and / or reduce the severity of one or more symptoms of the disease, disorder or condition. The person of ordinary skill in the art recognizes that the term “prevent” is not an absolute term. In the medical art it is understood to refer to the prophylactic administration of a drug to diminish the likelihood or seriousness of a disease, disorder or condition, or a symptom thereof, and this is the sense that such terms are used in this disclosure.

[0030] The phrase “targeting ligand” refers to a ligand that binds to “a targeted receptor” that distinguishes the cell being targeted from other cells. The ligands may be capable of binding due to expression or preferential expression of a receptor for the ligand, accessible for ligand binding, on the target cells. While use of a bioavailable nanoparticle including a human cell membrane may not require a separate targeting ligand, other types of bioavailable nanoparticles may benefit from the attachment of targeting ligands thereto. Examples of such ligands include any ligands relevant to VG health and protection, including but notlimited to avfl3 integrin ligand LXW7 (targeting avf33 integrin); avf33 integrin ligand GRGD peptide and E-selectin binding peptide.

[0031] The phrase “a targeted receptor” refers to a receptor expressed by a cell that is capable of binding a cell targeting ligand. The receptor may be expressed on the surface of the cell. The receptor may be a transmembrane receptor. Examples of such targeted receptors include galactose receptors expressed by hepatocytes, receptors for C-type lectins (e.g., mannose receptor) receptors expressed by macrophages, EGFR expressed by endothelial cells, and a variety of lectin receptors expressed by bacteria.

[0032] A “cell penetrating peptide” (CPP), also referred to as a “protein transduction domain” (PTD), a “membrane translocating sequence,” and a “Trojan peptide”, refers to a short peptide (e.g., from 4 to about 40 amino acids) that has the ability to translocate across a cellular membrane to gain access to the interior of a cell and to carry into the cells a variety of covalently and noncovalently conjugated cargoes, including proteins, oligonucleotides, and liposomes. They are typically highly cationic and rich in arginine and lysine amino acids. Examples of such peptides include TAT cell penetrating peptide (GRKKRRQRRRPQ); MAP (KLALKLALKALKAALKLA); Penetratin or Antenapedia PTD (RQIKWFQNRRMKWKK); Penetratin- Arg: (RQIRIWFQNRRMRWRR); antitrypsin (358-374): (CSIPPEVKFNKPFVYLI); Temporin L: (FVQWFSKFLGRIL-NH2);Maurocalcine: GDC(acm) (LPHLKLC); pVEC (Cadherin-5): (LLIILRRRIRKQAHAHSK); Calcitonin: (LGTYTQDFNKFHTFPQTAIGVGA P); Neurturin: (GAAEAAARVYDLGLRRLRQRRRLRRERVRA); Penetratin: (RQIKIWFQNR RMKWKKGG); TAT-HA2 Fusion Peptide: (RRRQRRKKRGGDIMGEWGNEIFGAIAGFLG); TAT (47-57) Y(GRKKRRQRRR);SynBl (RGGRLSYSRRRFSTSTGR); SynB3 (RRLSYSR RRF); PTD-4 (PIRRRKKLRRL); PTD-5 (RRQRRTSKLMKR); FHV Coat-(35-49) (RRRRNRTRRNRRRVR); BMV Gag-(7-25) (KMTRAQRRAAARRNRWTAR); HTLV-II Rex-(4-16) (TRRQRTRRARRNR); HIV-1 Tat (48-60) or D-Tat (GRKKRRQRRRPPQ); R9-Tat (GRRRRRRRRRPPQ); Transportan (GWTLNSAGYLLGKINLKALAALAKKIL chimera); SBP or Human Pl (MGLGLHLLVLAAALQGAWSQPKKKRKV); FBP (GALFLGWLGAAGS TMGAWSQPKKKRKV); MPG (ac- GALFLGFLGAAGSTMGAWSQPKKKRKV-cya (wherein cya is cysteamine));MPG(ANLS) (ac- GALFLGFLGAAGSTMGAW SQPKSKRKV-cya); Pep-1 or Pep-1- Cysteamine (ac-KETWWETWWTEWSQPKKKRKV-cya); Pep-2 (ac-KETWFETWFTEWSQPKKKRKV-cya); Periodic sequences, Polyarginines (RxN (4<N<17) chimera); Polylysines (KxN (4<N<17) chimera); (RAca)6R; (RAbu)6R; (RG)6R; (RM)6R; (RT)6R;(RS)6R; RIO; (RA)6R; and R7.

[0033] A “dye” refers to small organic molecules having a molecular weight (actual, not number average) of 2,000 Da or less or a protein which is able to emit light. Non-limiting examples of dyes include fluorophores, chemiluminescent or phosphorescent entities. For example, dyes useful in the present technology include but are not limited to cyanine dyes (e.g., Cy2, Cy3, Cy5, Cy5.5, Cy7, and sulfonated versions thereof), fluorescein isothiocyanate (FITC), ALEXA FLUOR® dyes (e.g., ALEXA FLUOR® 488, 546, or 633), DYLIGHT® dyes (e.g, DYLIGHT® 350, 405, 488, 550, 594, 633, 650, 680, 755, or 800) or fluorescent proteins such as GFP (Green Fluorescent Protein).

[0034] A “metal chelating ligand” as used herein refers to ligands that chelate metal isotopes for use in imaging. Non-limiting examples of metal chelating ligands include triazacyclononane-phosphinic acid (i.e., TRAP), 1,4,7, 10-tetraazacy clododecane- 1,4, 7,10- tetraacetic acid (i.e., DOTA), 1,4,7-triazacyclononane-triacetic acid (i.e., NOTA), diethylenetriaminepentaacetic acid i.e., DTP A), or chelating peptides). Thus, the metal chelating ligand may be one for use in PET or MRI.

[0035] In one aspect, the present technology provides methods of reducing vein graft injury comprising administering an effective amount of a bioavailable nanoparticle comprising NAD+and / or NADH to the vein to be grafted prior to surgical implantation of the vein graft in a subject. “Bioavailable nanoparticles comprising NAD+and / or NADH” of the present technology are synthetic nanoparticles (and do not include naturally occurring particles such as cells or organelles) that are capable of delivering NAD+and / or NADH across a cell membrane, e.g., an endothelial cell membrane, at a therapeutically effective level . In some embodiments, administering an effective amount of the bioavailable nanoparticle comprises storing the vein to be grafted in an aqueous solution comprising the effective amount of the bioavailable nanoparticle. Prior to or concurrently with storage in the aqueous solution, the vein graft may be flushed one or more times (e.g., once, twice, three times, four times, five times, or more) with the aqueous solution containing the effective amount of NAD+and / or NADH.

[0036] In the present methods of reducing vein graft injury, the aqueous solution may include saline and / or one or more additives selected from the group consisting of potassium lactobionate, potassium hydrogen phosphate, magnesium sulfate, raffinose, denosine, glutathione, allopurinol, and hydroxyethyl starch. The aqueous solution may be kept at room temperature or may be cooled, e.g., in an ice bath or using a chiller. Thus, in some embodiments of the present methods, the aqueous solution may have a temperature from 0°C to about 25°C, including 0°C or any of about 1°C, 2°C, 3°C, 4°C, 5°C, 6°C, 7°C, 8°C, 9°C, 10°C, 11°C, 12°C, 13°C, 14°C, 15°C, 16°C, 17°C, 18°C, 19°C, 20°C, 21°C, 22°C, 23°C, 24°C, 25°C or a range between and including any two of the foregoing values. For example, the aqueous solution may be kept at a temperature from 0°C to about 5°C.

[0037] In some embodiments of the present methods, the effective amount of NAD+and / or NADH may be about 1 pM to about 10 mM. For example, the effective amount of NAD+and / or NADH may be about 1 pM, 2 pM, 3 pM, 4 pM, 5 pM, 10 pM, 15 pM, 20 pM, 25 pM, 30 pM, 40 pM, 50 pM, 60 pM, 70 pM, 80 pM, 90 pM, 100 pM, 125 pM, 150 pM, 200 pM, 300 pM, 400 pM, 500 pM, 750 pM, 1 mM, 2 mM, 5 mM, 10 mM or a range between and including any two of the foregoing values. For example, the effective amount of NAD+and / or NADH may be about 2 pM to about 100 pM or from about 5 pM to about 20 pM.

[0038] In another aspect, the present technology provides methods of improving vein graft health and nanoparticles for same. The methods include administering an effective amount of a bioavailable nanoparticle comprising NAD+and / or NADH and a coating comprising a human cell membrane to a subject post-surgical implantation of the vein graft in the subject, wherein the human cell membrane is selected from the group consisting of a human platelet membrane, a human red blood cell membrane, a human white blood cell membrane, mesenchymal stem cell membrane, and a human macrophage cell membrane. In any embodiments of the present methods, the coating comprises a human platelet membrane. The bioavailable nanoparticles may include about 30 wt% to about 75 wt% human cell membrane, e.g., any of about 30 wt%, 31 wt%, 32 wt%, 33 wt%, 34 wt%, 35 wt%, 40 wt%, 45 wt%, 50 wt%, 55 wt%, 60 wt%, 65 wt%, 70 wt%, 75 wt% human cell membrane or a range between and including any two of the foregoing values. Thus, in some embodiments,the bioavailable nanoparticles include about 30 wt% to about 70 wt%, about 30 wt% to about 40 wt%, about 40 wt% to about 60 wt%, about 45 wt% to about 55 wt%, or about 50 wt%.

[0039] In still another aspect, the present technology provides methods of preventing or treating post-operative restenosis comprising administering an effective amount of a bioavailable nanoparticle comprising NAD+and / or NADH and a coating comprising a human cell membrane to a subject after surgical reconstruction of a blood vessel in the subject, wherein the human cell membrane is selected from the group consisting of a human platelet membrane, a human red blood cell membrane, a human white blood cell membrane, mesenchymal stem cell membrane, and a human macrophage cell membrane. In some embodiments, the surgical reconstruction of the blood vessel in the subject comprises balloon angioplasty and / or placement of a stent. In any embodiments of the present methods, the coating comprises a human platelet membrane.

[0040] In the methods of improving vein graft health or of preventing or treating postoperative restenosis, the effective amount of NAD+and / or NADH may vary with the application but typically falls within the range of about 0.1 mg / kg (of the subject being treated) to about 200 mg / kg. Thus, in some embodiments the effective amount may be any of about 0.1, 0.2, 0.5, 1, 2, 3, 4, 5, 6, 7. 8, 9, 10, 11, 12, 13, 14, 15, 20, 25, 30, 40, 50, 60, 70, 80, 90, 100, 125, 150, 175, or 200 mg / kg or a range between and including any two of the foregoing values. For example, the effective amount of NAD+and / or NADH may be about 1 mg / kg to about 100 mg / kg or about 5 mg / kg to about 20 mg / kg.

[0041] A variety of bioavailable nanoparticles may be used to deliver the NAD(H) in methods of the present technology. For example, the bioavailable nanoparticles may include an inorganic core and NAD+and / or NADH, and a coating including a lipid bilayer. The inorganic core may be calcium phosphate or a metal organic framework (MOF). The MOF may include a transition metal ion coordinated to a coordinating ligand, wherein the transition metal ion is selected from the group consisting of zinc, iron, zirconium, copper, and cobalt ions, and the coordinating ligand is selected from an imidazolate ligand or a carboxylate ligand. While not wishing to be bound by theory, the NAD(H)-loaded NPs are belived to be taken up by the cells via endocytosis and directly replenish cellular NAD+.The CaP or MOF cores are believed to dissolve in the acidic environment of the endosome, leading to endosome swelling and bursting (due to an increase in osmotic pressure) to release the entrapped payload into cytosol.

[0042] The amount of NAD+orNADH loaded into the nanoparticle may vary, e.g., from 1 wt% to 50 wt% NAD+, NADH, or both. Thus, in any embodiments, the nanoparticles of the present methods may include 1 wt%, 2 wt%, 3 wt%, 4 wt%, 5 wt%, 6 wt%, 7 wt%, 8 wt%, 9 wt%, 10 wt%, 11 wt%, 12 wt%, 13 wt%, 14 wt%, 15 wt%, 20 wt%, 25 wt%, 30 wt%, 35 wt%, 40 wt%, 45wt%, or 50 wt% NAD+and / or NADH, or a range between and including any two of the foregoing values, e.g., 1 wt% to 25 wt% or 2 wt% to 20 wt%, or 3 wt% to 15 wt% NAD+and / or NADH among others.

[0043] In any embodiments of the present methods where the NPs include a lipid bilayer, any suitable lipids may be used. For example, the lipids of the lipid bilay er(s) may be selected from the group consisting of L-a-phosphatidylcholine (PC), 1,2-dioleoyl-sn- glycero-3 -phosphate (DOPA), l,2-dioleoyl-3 -trimethylammonium -propane (DOTAP), 1,2- dioleoyl-sn-glycero-3 -phosphocholine (DOPC), l,2-dioleoyl-sn-glycero-3- phosphoethanolamine (DOPE), l,2-dioleoyl-sn-glycero-3-phospho-L-serine (DOPS), 1,2- distearoyl-sn-glycero-3-phosphorylethanolamine (DSPE), l,2-distearoyl-sn-glycero-3- phosphoethanolamine-N-[methoxy(poly ethylene glycol)] (DSPE-PEG), 1,2-dimyristoyl- rac-glycerol (DMG), l,2-dimyristoyl-rac-glycero-3-methoxypolyethylene glycol (DMG- PEG), cholesterol, and combinations of two or more thereof. In any embodiments of the NPs, the lipids of the lipid bilayer may include a combination of PC and / or DOPA. In any embodiments, the lipids of the lipid bilayer may include a combination of PC and / or DOPA, and cholesterol. The lipids of the lipid bilayer may include DSPE and / or DSPE-PEG, and / or DMG and / or DMG-PEG.

[0044] In any embodiments of the present methods, a portion of lipids in the lipid bilayer of the NPs may be conjugated to polyethylene glycol) (PEG). Up to 100 mol% of the lipids in the lipid bilayer may be conjugated to PEG, e.g., 1, 2, 3, 4, 5, 6, 7, 8, 10, 15, 20, 30, 40, 50, 60, 70, 80, 90, or 100 mol%, or a range between and including any two of the foregoing values. In any embodiments, the lipids of the lipid bilayer may include DSPE- PEG. The PEG (when conjugated to a lipid) includes a free terminus selected from thegroup consisting of OH, O-C1-4 alkyl ether, NH2, NHR, COOH, COOR, wherein R is an alkyl or alkenyl group, a dye, a targeting ligand, and a metal chelating ligand. Thus, the PEG terminus may be optionaly conjugated to a dye, a targeting ligand, or a metal chelating ligand directly or through any suitable linker (e.g., with a molecular weight below about 500 Da) known in the art. The PEG may have a number average molecular weight ranging from 300 to 10000 Da. In any embodiments the PEG may have a number average molecular weight of about 300, 500, 1000, 2000, 3000, 4000, 5000, 6000, 7000, 8000, 9000, or 10000 Da or a range between and including any two of the foregoing values, e.g., 3000-7000 Da. In any embodiments, the lipid of the lipid bilayer(s) includes a cell membrane extracted from a red blood cell, macrophage, neutrophil, or platelet, and combinations of two or more thereof.

[0045] In any embodiments, the nanoparticles of the present methods may include lipid bilayer(s) which coat the surface of the nanoparticle in whole or in part. The nanoparticles may include, e.g., 10 wt% to 75 wt% lipid bilayer(s). Thus, in any embodiments, the present nanoparticles may include 10 wt%, 15 wt%, 20 wt%, 25 wt%, 30 wt%, 35 wt%, 40 wt%, 45 wt%, 50 wt%, 55 wt%, 60 wt%, 65 wt%, 70 wt%, 75 wt% lipid bilayer(s), or a range between and including any two of the foregoing values, e.g., 10 wt% to 40 wt%, 10 wt% to 30 wt%, 15 wt% to 40 wt%, 20 wt% to 75 wt%, 30 wt% to 70 wt%, or 50 wt% to 70 wt% lipid bilayer(s).

[0046] In any embodiments of the present methods where the bioavailable NPs include a lipid bilayer, the lipid bilayer may include or be encapsulated by (in part or in whole) a human cell membrane selected from the group consisting of a human platelet membrane, a human red blood cell membrane, a human white blood cell membrane, mesenchymal stem cell membrane, and a human macrophage cell membrane. For example, the lipid bilayer may include or be encapsulated by (in part or in whole) a human platelet membrane. In any embodiments, the weight ratio of of the human cell membrane coating to the remainder of the bioavailable nanoparticle (e.g., inorganic core, lipid bilayer, and NAD(H)) may be 2: 1 to 1 :2. In some embodiments, the weight ratio of of the human cell membrane coating to the remainder of the bioavailable nanoparticle (e.g., inorganic core, lipid bilayer, and NAD(H)) may be about 1 : 1. In some embodiments, the human cell membrane coating may be ahuman platelet membrane coating, and the weight ratio of of the human platelet membrane coating to the remainder of the bioavailable nanoparticle (e.g., inorganic core, lipid bilayer, and NAD(H)) may be, e.g., about 2: 1 to about 1 :2 or, e.g., about 1 : 1. In any embodiments

[0047] In any embodiments, cell penetrating peptides, targeting ligands, and / or metal chelating agents may optionally be conjugated directly or indirectly (through any suitable linker of less than 500 Da known in the art) to lipids in the lipid bilayer.

[0048] In any embodiments, the present NPs include 10 wt% to 90 wt% inorganic core, e.g., 10 wt% - 90 wt% calcium phosphate or 10 wt% - 90 wt% MOF. For example, the present nanoparticles may include 10 wt%, 15 wt%, 20 wt%, 25 wt%, 30 wt%, 35 wt%, 40 wt%, 45 wt%, 50 wt%, 55 wt%, 60 wt%, 65 wt%, 70 wt%, 75 wt%, 80 wt%, 85 wt%, 90 wt% inorganic core or a range between and including any two of the foregoing values, e.g., 20 wt% to 80 wt%, or 40 wt% to 60 wt%.

[0049] In any embodiments, the inorganic core is calcium phosphate. Calcium phosphate may be a single compound but is more typically a mixture of two or more such compounds having, e.g., a molar ratio of Ca to P from 0.5 to 2 (e.g., 0.5, 1.2, 1.33, 1.5, 1.67, 2 or a range between and including same). In some embodiments the calcium phosphate comprises one or more of hydroxy apatite (Caio(P04)e(OH)2), Ca(H2PO4)2*H2O, Ca(H2PO4)2, CaHPO4*2H2O, Ca(HPO4) and the like.

[0050] In any embodiments, the inorganic core of the present nanoparticles may be a MOF. As noted above the transition metal ion of the MOF may be selected from the group consisting of zinc, iron, zirconium, copper, and cobalt ions. In any embodiments the transition metal ion of the MOF may be zinc ions or iron ions. The coordinating ligand may be an imidazolate ligand or a carboxylate ligand as noted above. Imidazolate ligands are coordinating ligands that contain an imidazole group such as, e.g., imidazole itself, 2- methyl-imidazole, benzimidazole, or 5-methylbenzimidazole. Carboxylate ligands include, e.g., terephthalic acid, 2-methyl-pterphthalic acid, 2 -hydroxy -terphthalic acid, and 2-amino- terphthalic acid. The imidazolate and carboxylate ions are typically but are not necessarily in their anionic forms. Those of skill in the art will recognize which ligands are suitable for use with a particular type of metal to form a metal organic framework component. By wayof example only, zinc may be used with imidazolate ligands and iron may be used with carboxylate ligands, especially dicarboxylate ligands. In any embodiments, the coordinating ligand also may be selected from the group consisting of imidazole, 2-methyl- imidazole, benzimidazole, 5 -methylbenzimidazole, terephthalic acid, 2-methyl-pterphthalic acid, 2-hydroxy -terphthalic acid, and 2-amino-terphthalic acid, benzene- 1, 3, 5-tricarboxylic acid, l,3,5-tris(4-carboxyphenyl)benzene, 2,6-naphthalenedicarboxylic acid, 4,4',4"-s- triazine-2,4,6-triyl-tribenzoic acid, and 2,5-dihydroxyterephthalic acid. In any embodiments, the coordinating ligand is 2-methyl-imidazole. In any embodiments, the the MOF includes zinc ions and imidazolate ligands. In any embodiments, the imidazolate ligand may be selected from imidazole, 2-methyl-imidazole, benzimidazole, or 5- methylbenzimidazole. In any embodiments of the present NPs, the imidiazolate ligand may be 2-methyl-imidazole.

[0051] The present nanoparticles may include a metal organic framework component as described above. Suitable metal ions that may be employed in the metal organic framework component include zinc, iron, zirconium, copper, and cobalt ions. In any embodiments the metal ion may be zinc ion or it may be iron ion. The coordinating ligand may be an imidazolate ligand or a carboxylate ligand as noted above. Imidazolate ligands are coordinating ligands that contain an imidazole group such as, e.g., imidazole itself, 2- methyl-imidazole, benzimidazole, or 5-methylbenzimidazole. Carboxylate ligands include, e.g., terephthalic acid, 2-methyl-pterphthalic acid, 2-hydroxy -terphthalic acid, and 2-amino- terphthalic acid. The imidazolate and carboxylate ions are typically but are not necessarily in their anionic forms. Those of skill in the art will recognize which ligands are suitable for use with a particular type of metal to form a metal organic framework component. By way of example only, zinc may be used with imidazolate ligands and iron may be used with carboxylate ligands, especially dicarboxylate ligands.

[0052] The present nanoparticles may have a hydrodynamic diameter ranging from at least 50 nm to less than 1000 nm. For example, the NPs may have a hydrodynamic diameter of 50, 60, 70, 80, 90, 100, 110, 130, 150, 170, 200, 250, 300, 400, 500, 600, 700, 800, 900, or less than 1000 nm or a range between and including any two of the foregoing values. Such ranges include but are not limited to NPs with a hydrodynamic diameter of 70to 700 nm or 100 to 400 nm. Alternatively, the present technology provides NPs with a median or average hydrodynamic diameter, also selected from any of the foregoing values or ranges.

[0053] In any embodiments, the bioavailable nanoparticle may be a nanoparticle comprising a disulfide-containing lipopeptide as disclosed in USSN 63 / 443,675 (filed 2 / 6 / 2023 and titled “Disulfide-containing lipopeptides, nanoparticles and methods of use”), incorporated by reference herein and for all purposes. Thus, the bioavailable nanoparticle may include a compound of Formula I:I a stereoisomer thereof, and / or a pharmaceutically acceptable salt thereof, whereinX1and X2are independently absent or selected from unsubstituted Ci-6 alkylene or C2-6 alkenylene;X3is selected from unsubstituted C1-6 alkylene or C2-6 alkenylene;Y1and Y2, are each independently absent, C(O)O, or C(O)NH;Y3is absent, C(O), C(O)O, or C(O)NH;Y4is C(O)O, C(O)NH, NHC(O)O, or NHC(0)NH;R1and R2are independently selected from unsubstituted Cs-24 alkyl or Cs-24 alkenyl groups;R3and R4are independently absent or selected from an amino acid residue, a peptide or isopeptide comprising 2-10 amino acid residues, or a C1-12 alkyl group, each ofwhich is optionally substituted with 1, 2, or 3 ionizable functional groups such that at least one ionizable functional group is present on at least one of R3and R4, provided that Y3is absent when R3is an amino acid, peptide, or isopeptide, and Y4is absent when R4is an amino acid residue, peptide, or isopeptide; and further provided that if one of R3and R4is absent, the other is present; and n and p are each independently 1, 2, 3, 4, or 5.

[0054] In one embodiment, the present technology provides a bioavailable nanoparticle that includes a compound of Formula I that is a compound of Formula IA,a stereoisomer thereof, and / or a pharmaceutically acceptable salt thereof, whereinX1and X2are independently absent or selected from unsubstituted Ci-6 alkylene or C2-6 alkenylene;X3is selected from unsubstituted C1-6 alkylene or C2-6 alkenylene;Y1and Y2are each independently selected from C(O)O or C(O)NH;R1and R2are independently selected from unsubstituted Cs-24 alkyl or Cs-24 alkenyl groups;R3and R4are independently selected from C1-10 alkyl groups substituted with 1, 2, or 3 ionizable functional groups; and n and p are each independently 1, 2, 3, 4, or 5.

[0055] In any embodiments, the bioavailable nanoparticle includes a compound of Formula I (or IA) that is a compound of Formula II,

[0056] In any embodiments, bioavailable nanoparticle includes a compound of Formula I (or IA) that is a compound of Formula III,III

[0057] In any embodiments, the present compounds may include 1, 2, or 3 ionizable functional groups. In any embodiments, each ionizable functional group may be independently selected from NH2, NHR, NR2, guanidine, imidazole, or amidine, wherein each R is independently an unsubstituted C1-6 alkyl, phenyl, or benzyl group. In any embodiments, at least one ionizable functional group is guanidine.

[0058] Thus, the bioavailabl nanoparticle includes any of the disulfide-containing lipopeptides disclosed herein, including but not limited to compounds of Formulas I, II, and III. In any embodiments, the bioavailable nanoparticle may further include a PEG-lipid, e.g., PEG-DSPE and / or PEG-DMG. In any embodiments, the nanoparticles may further include a structural lipid, e.g., cholesterol and / or P-sitosterol. In any embodiments, the nanoparticles may further include a phospholipid, e.g., DSPC and / or DOPE.

[0059] In any embodiments, the bioavailable nanoparticle may be a nanoparticle comprising a poly(amidoamine) oligomer, as disclosed in USSN 63 / 546,129 (filed 10 / 27 / 2023 and titled “Lipid nanoparti cis formed by lipidoids for efficient delivery of nucleotide drugs and biologies”), incorporated by reference herein and for all purposes. Thus, the bioavailable nanoparticle may include in one aspect, the present technology provides a compound of Formula IV,IV whereinPAO is a linear or branched polyamine or poly(amidoamine) oligomer, each having 2 to 32 amine groups and 2 to 100 carbon atoms, wherein each ester side-chain is connected to the PAO by one of the amine groups;R at each occurrence is independently selected from a Ce-20 alkyl group, a Ce-20 alkenyl group, or TG;TG is a monosaccharide selected from the group consisting of glucose, O-protected glucose, galactose, O-protected galactose, fructose, O-protected fructose, O-protected N-acetylgalactosamine, and N-acetylgalactosamine; and n is 2 to 32.

[0060] The PAO of structure IV may be a linear or branched polyamine or poly(amidoamine) oligomer having 2 to 32 amine groups, i.e., 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 32 amine groups or a range between any two of the foregoing values. For example, the PAO may have 4 to 30, 5 to 28, 6 to 26, or 8 to 26 amine groups. The PAO may have 2 to 120 carbon atoms, e.g., 2, 3, 4, 5, 6, 7, 8, 9, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 105, 110, 115, 120 or a range between any two of the foregoing values. For example, the PAO may have 4 to 120 carbon atoms, 6 to 120 carbon atoms, 8 to 110 carbon atoms, 4 to 100 carbon atoms, 6 to 100 carbon atoms, or 10 to 100 carbon atoms. In any embodiments the PAO may be linear. In any embodiments, the PAO may be branched. In any embodiments, the PAO may have a carbon chain of 2-4 carbons connecting a pair of amine groups. The carbon chain of 2-4 carbons may be a linear alkylene group. In any embodiments, each pair of amine groups may be connected by a carbon chain of 2-4 carbons. In any embodiments, the PAO may have 6 to 24 or 8 to 26 amine groups. In any embodiments, the PAO may have 10 to 100 carbon atoms. In any embodiments, the PAO may have 10 to 16 amine groups and 18 to 60 carbon atoms. In any embodiments, the PAO may be branched.

[0061] In the compound of Formula IV, n may also be 2 to 32, i.e., any of 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, or 32 or a range between and including any two of the foregoing values. For example, n may be 4 to 8 or

[0062] In any embodiments, the PAO may be a linear polyamine oligomer. In any embodiments, the PAO may be a branched polyamine oligomer, e.g., a polyethyleneimine (PEI) oligomer. In any embodiments, the PAO (including but not limited to the PEI oligomer) may have a number average molecular weight of about 200 Da to about 1800 Da, e.g., any of about 200, 400, 600, 800, 1000, 1200, 1400, 1600, 1800 or a range between and including any two of the foregoing values. Thus, in any embodiments, the PAO may have a number average molecular weight of about 400 Da to about 800 Da.

[0063] In any embodiments, the PAO may comprise a poly(amidoamine) oligomer(PAMAM), e.g., a PAMAM dendrimer with an ethylenediamine core. In any embodiments,the PAMAM dendrimer may be a generation 0 or 1. In any embodiments, the PAMAM dendrimer may be capped with a C2-4 alkylenediamine, e.g., ethylenediamine. Thus, in any embodiments, the PAO may have the structure of Formula V:

[0064] It will be understood by those skilled in the art that the squiggly lines indicate the attachment points to the ester side chains of Formula IV.

[0065] As a small polymer, the PAO may or may not be polydisperse. Typically, the PAO may have a poly dispersity of about 1 to about 2. For example, the poly dispersity may be any of about 1, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, 2, or may be within a range between and including any two of the foregoing values, such as about 1 to about 1.5.

[0066] In any embodiments of the compound of Formula IV, at least one occurrence of R is a Ce-20 alkylene group. In any embodiments at least one occurrence of R is a Cio-is alkylene group. In any embodiments, at least one occurrence of R is an Ce-20 alkenyl group. In some such embodiments, the Ce-20 alkenyl group may have 1, 2 or 3 carbon-carbon double bonds. In any embodiments, at least one occurrence of R is a Cio-is alkenyl group having 1 or 2 carbon-carbon double bonds. In any embodiments, at least one occurrence of R is TG. In any embodiments, TG may be glucose, galactose, fructose, or N- acetylgalactosamine. In any embodiments, TG is glucose. In any embodiments, TG may be O-protected galactose, fructose, O-protected fructose, or O-protected N-acetylgalactosamine. In some such embodiments, the O-protected moieties are protected as ketals, e.g., acetonides.

[0067] The compound of Formula IV may include various amounts of different R groups. For example, about 10% to about 90% or about 25% to about 75% of R groups may be Ce-20 alkyl groups (e.g., Cio-is alkyl groups or any of those disclosed herein), and the remaining R groups may be TG. In any embodiments, about 25% to about 50% of R groups may be a Ce-20 alkyl group (e.g., Cio-is alkyl groups or any of those disclosed herein) and the remaining R groups may be TG. In any embodiments, about 50% of R groups may be Ce-20 alkyl groups, e.g., Cio-is alkyl groups or any of those disclosed herein.

[0068] Thus, the present technology provides lipid nanoparticles (LNP) that include a compound of Formula IV as disclosed herein and a PEG-lipid. In any embodiments, the LNP includes about 75 wt% to about 95 wt% of the compound of Formula IV and about 5 wt% to about 25 wt% of the PEG-lipid. In any embodiments, the LNP includes about 85 wt% to about 90 wt% of the compound and about 10 wt% to about 15 wt% of the PEG- lipid.

[0069] LNPs of the present technology may include a variety of PEG-lipids. For example, the LNP may include DMG-PEG, DSPE-PEG and / or ceramide-PEG. In any embodiments, the PEG-lipid may include DMG-mPEG. The PEG-lipid may further include CPP-DMG- mPEG and / or glucose-DMG-mPEG.

[0070] A pharmaceutical composition comprising bioavailable nanoparticle as described herein and a pharmaceutically acceptable carrier and / or excipient. In any embodiments of the present methods, the bioavailable nanoparticles may be administered to the subject, e.g., a human subject, as a pharmaceutical composition.

[0071] The compositions described herein can be formulated for various routes of administration, for example, by injection, e.g., intravenous injection, or by parenteral, intravitreal, intrathecal, intracerebroventricular, rectal, nasal, vaginal administration, or via a temporary and / or implanted reservoir (e.g., a stent comprising a reservoir of bioavailable nanoparticles. Parenteral or systemic administration includes, but is not limited to,subcutaneous, intravenous, intraperitoneal, and intramuscular injections. The following dosage forms are given by way of example and should not be construed as limiting the instant present technology.

[0072] Injectable dosage forms generally include solutions or aqueous suspensions which may be prepared using a suitable dispersant or wetting agent and a suspending agent so long as such agents do not interfere with formation of the nanoparticles described herein. Injectable forms may be prepared with acceptable solvents or vehicles including, but not limited to sterilized water, phosphate buffer solution, Ringer's solution, 5% dextrose, or an isotonic aqueous saline solution.

[0073] Besides those representative dosage forms described above, pharmaceutically acceptable excipients and carriers are generally known to those skilled in the art and are thus included in the instant present technology. Such excipients and carriers are described, for example, in “Remingtons Pharmaceutical Sciences” Mack Pub. Co., New Jersey (1991), which is incorporated herein by reference. Exemplary carriers and excipients may include but are not limited to USP sterile water, saline, buffers (e.g., phosphate, bicarbonate, etc.), tonicity agents (e.g., glycerol),

[0074] Specific dosages may be adjusted depending on conditions of disease, the age, body weight, general health conditions, sex, and diet of the subject, dose intervals, administration routes, excretion rate, and combinations of drug conjugates. Any of the above dosage forms containing effective amounts are well within the bounds of routine experimentation and therefore, well within the scope of the instant present technology. By way of example only, such dosages may be used to administer effective amounts of the present bioavailable nanoparticles to the patient and may include 0.1, 0.2, 0.5, 1, 2, 3, 4, 5, 6, 7. 8, 9, 10, 11, 12, 13, 14, 15, 20, 25, 30, 40, 50, 60, 70, 80, 90, 100, 125, 150, 175, or 200 mg / kg or a range between and including any two of the forgoing values such as about 0.1 to about 200 mg / kg, or about 1 mg / kg to aboutlOO mg / kg, or about 5 mg / kg to about 20 mg / kg. Such amounts may be administered parenterally as described herein and may take place over a period of time including but not limited to 5 minutes, 10 minutes, 20 minutes, 30 minutes, 45 minutes, 1 hour, 2 hours, 3 hours, 5 hours, 10 hours, 12, hours, 15 hours, 20 hours, 24 hours or a range between and including any of the foregoing values. Thefrequency of administration may vary, for example, once per day, per 2 days, per 3 days, per week, per 10 days, per 2 weeks, or a range between and including any of the foregoing frequencies. More frequent administration is also possible. Alternatively, the compositions may be administered once per day on 2, 3, 4, 5, 6 or 7 consecutive days. A complete regimen may thus be completed in only a few days or over the course of 1, 2, 3, 4 or more weeks.

[0075] The examples herein are provided to illustrate advantages of the present technology and to further assist a person of ordinary skill in the art with preparing or using the nanoparticles compositions of the present technology. To the extent that the compositions include ionizable components, salts such as pharmaceutically acceptable salts of such components may also be used. The examples herein are also presented in order to more fully illustrate the preferred aspects of the present technology. The examples should in no way be construed as limiting the scope of the present technology, as defined by the appended claims. The examples can include or incorporate any of the variations or aspects of the present technology described above. The variations or aspects described above may also further each include or incorporate the variations of any or all other variations or aspects of the present technology.EXAMPLESMaterials and General Procedures

[0076] Materials. P-nicotinamide adenine dinucleotide (NAD+), calcium chloride (CaCh), and IGEPAL CO-520 were purchased from Sigma-Aldrich (St. Louis, MO, USA). Disodium hydrogen phosphate (Na2HPO4) was purchased from Dot Scientific Inc., (Burton, MI, USA). Dioleoyhosphatydic acid (DOPA) and L-a-phosphatidylcholine (Soy PC) were obtained from Avanti Polar Lipids (Alabaster, AL, USA). ATTO-550 tracrRNA was bought from Integrated DNA Technologies (Coralville, IA, USA). Cyclohexane and chloroform were purchased from Thermo Fisher Scientific (Fitchburg, WI, USA).

[0077] Characterization. Dynamic light scattering (DLS) was used to measure the hydrodynamic diameter and zeta potential of P-NAD+-NP and related nanoparticles using aZetaSizer Nano ZS90 spectrometer (Malvern Instruments, USA). Transmission electron microscopy (TEM, FEI Tecnai G2 F30 TWIN 300 KV, E.A. Fischione Instruments, Inc., USA) was used to determine the morphology of these nanoparticles. To evaluate the loading efficiency and loading content of the NAD(H)-loaded NTs, the NPs were dissolved in 0.01 M HC1 to release NAD(H). The pH of the solutions was adjusted to neutral, and the NAD(H) concentration was determined by HPLC or an Amplite™ Colorimetric Total NAD and NADH Assay Kit (AAT Bioquest, Sunnyvale, CA, USA). For the HPLC method, mobile phase A (20 raM NalUPC ) and mobile phase B (acetonitrile / acetic acid 0.01%) were delivered in a gradient elution mode at 1 mL / min. Phase B increased linearly from 0% to 60% from 0 to 10 min. The NAD(H) signals were detected at 260 nm.

[0078] Human Vein Explant Study. An ex vivo culture of completely de-identified human great saphenous vein explants was used to mimic human physiology in an experimental system in accordance with our established methodologies using human VG explants to study EC physiology and IH.21'23Fresh human great saphenous veins (acquired from excess grafts for VG bypass procedures) were immediately segmented into rings of ~0.5cm in length and divided into individual wells of 24-well culture plates containing normal saline supplemented with or without 10 pM NAD+-NP. To recapitulate the intraoperative VG storage conditions, the covered 24-well culture plates containing VG explants were left in ambient environments for 6hr, followed by downstream analysis.

[0079] Animal Protocol. All animal studies conform to the National Institutes of Health Guide for the Care and Use of Laboratory Animals and The ARRIVE guidelines (Animal Research: Reporting of In Vivo Experiments). Animal research protocol was approved by the Animal Care and Use Committee (ACUC) at University of Virginia.

[0080] Rat Autologous Vein Graft Bypass Procedure. Rat autologous vein graft bypass procedure was performed using a cuff-assisted, external jugular vein-to-common carotid artery interposition graft model. Briefly, male Sprague-Dawley (SD) rats (300-350g) were anesthetized with isoflurane, followed by external jugular vein isolation. After ligation of all side branches, an 8mm segment of the external jugular vein was excised and temporarily placed in normal saline for 3hr to mimic the intraoperative vein graft storage of a typical coronary or peripheral artery bypass graft procedure in human patients. Following theautologous vein collection, the midline incision was temporarily closed with metal surgical clips, and animals were properly recovered with the necessary analgesia and supporting treatment. Upon completion of the ex vivo intraoperative vein storage, the common carotid artery was exposed, ligated in the middle, and then severed with a micro scissor to create the distal and proximal ends for graft bypass. Each end of the common carotid artery was then clamped with micro bulldog clamp and then overwrapped with custom-made cuffs (adapted from BD 20G catheters). Both ends of now cuff-surrounded carotid artery were everted over the cuff and then fastened with suture. Following the aforementioned ex vivo incubation, the autologous jugular vein was sleeved over both everted arteries on cuffs and then secured with sutures. Upon recovered from anesthesia, animals were provided with ACUC-approved analgesic plan and supporting treatment. Animals were subjected to monthly non-invasive ultrasound imaging to determine the VG luminal patency.

[0081] mRNA Extraction and real-time quantitative polymerase chain reaction(qPCR). mRNA from human saphenous vein explant tissue homogenates or from primary culture of human aortic SMC and EC, as well as rat carotid artery tissue homogenates was isolated using TRIzol according to manufacturer's manual. Following quantitation using NanoDrop, 1 pg of mRNA was used for the synthesis of first-strand cDNA using the High- Capacity cDNA Reverse Transcription Kit (ThermoFisher Scientific, Waltham, MA). cDNA templates were then amplified in triplicates with SYBR Green PCR Master Mix using the QuantStudio3 real-time qPCR system (Applied Biosystems, Carlsbad, CA). The sequences for each primer set are provided in Table 1.

[0082] Balloon Angioplasty Procedure in Rat Carotid Arteries. Balloon Angioplasty for common carotid arteries was performed in male Sprague-Dawley rats (350g-380g) as described previously. In brief, the rats were anesthetized with isoflurance (5% for inductionand 2-3% for maintenance). After preparing the skin, a midline neck incision was made to expose the left common carotid artery. The branches of the artery were looped to avoid reflux of blood. To injure the artery, we inserted a 2-F Fogarty arterial embolectomy catheter (Edward Life Science, Irvine, CA) through an arteriotomy on the external artery. The balloon catheter was inflated at 1.5 atm and then withdrawn until the carotid bifurcation. After three times repeating this action followed by a fourth cycle with concomitant rotation of catheter during withdrawing, the catheter was removed. The external carotid artery was ligated permanently and the blood flow was resumed. Throughout the procedure, the rat was kept anesthetized via isoflurane, inhaling at a flow rate of 2-4L / min. Carprof en (5mg / kg) and Bupivacaine were subcutaneously injected.

[0083] Right after the procedure, we provided a single-dose regimen via intravenous injection with the following treatments: saline, empty biomimetic CaP-NP (empty NP), naked NAD+solution (free NAD+), and NAD+-loaded, biomimetic CaP-NP (P-NAD+-NP) — all at a dose equivalent to 10 mg / kg NAD+payload.

[0084] The common carotid arteries were collected two weeks after the procedure. During the harvest, the animals were anesthetized with isoflurane and the arteries were collected following perfusion fixation at a physiological pressure of lOOmmHg. Animals were euthanized in a chamber gradually filled with CO2.

[0085] Evans blue staining for re-reendothelialization evaluation. Balloon Angioplasty for common carotid arteries was performed in the same way as described above. 7 days after the procedure, re-endothelialization of the arteries was evaluated using an Evans blue staining assay as previously reported. 1 mL of 0.5% Evans blue solution (Sigma-Aldrich, St. Louis, MO) was injected through the tail vein. After 30 mins of injection, the rat was anesthetized as described above. The common carotid artery was collected following saline and RNAlater perfusion. Animals were euthanized in a chamber gradually filled with CO2.

[0086] Histological and Morphometric Analysis. Intimal hyperplasia: Paraffin cross sections (5 pm thick) were cut at equally spaced intervals using a microtome (Leica) and then stained (Hematoxylin and Eosin, H&E) for morphometric analysis of intimalhyperplasia, as described previously. Morphometric parameters were measured on the sections and calculated by using ImageJ software: lumen area, the area inside internal elastic lamina (IEL area), the area inside external elastic lamina (EEL area), intima area (= IEL area - lumen area), and media area (=EEL area - IEL area). Intimal hyperplasia (IH), the ratio of intima area versus media area (I / M) was calculated by (IEL area - lumen area) / (EEL area - IEL area). 3-6 sections from each rat were measured, and the measurement was pooled from all sections to generate a mean value for each animal. The mean values from all the animals in each treatment group were averaged, and then the standard error of the mean (SEM) was calculated.

[0087] Re-endothelialization evaluation: The common arteries were longitudinally opened for image acquisition. Re-reendothelialization area was measured and calculated using ImageJ software: the blue staining area (no endothelium coverage) and no-staining area (with endothelium coverage). Base level endothelium coverage was determined by arteries collected immediately after the procedure and was subtracted from Day 7 values. Re- reendothelialization (%) = (No-staining area - baseline) / (Blue staining area+ No-staining area - baseline).

[0088] All the measurements were performed by an independent researcher who was blinded to the experimental treatment.

[0089] Cell Culture. Primary culture of human aortic SMC (HASMC), human aortic EC (HAEC), and their respective optimal culture media (SmBm2 / EBM2 basal medium for experiments, and SmGm2 / EGM2 complete medium for expansion) were purchased from Lonza (Walkersville, MD). Primary cells between passage 4-8 were cultured at 37°C with 5% CO2 as previously described.5,6For subculture, Trypsin-EDTA and Accutase (Thermo Fisher Scientific, Waltham, MA) were used for cell detachment of HASMC and HAEC, respectively, as established in our prior reports.5,6

[0090] Statistical Analysis. Statistical analysis was performed using GraphPad Prism 9 and 10. Prior to parametric analysis, the normality and equal variance of data sets were tested. For two-group comparisons, Student’s t-test was performed. For multiple group-wisecomparisons, One-way analysis of variance (ANOVA) was performed followed by appropriate post-hoc analysis.Example 1 - Preparation of NAD+-lipid-calcium phosphate nanoparticles (NAD+-NP) and biomimetic platelet membrane-coated NAD+NPs (P-NAD+-NP)

[0091] NAD+-NP were synthesized utilizing a water-in-oil microemulsion method, followed by thin-film hydration, as described2. Briefly, two reverse water-in-oil microemulsions, each of 25 ml, were prepared in a cyclohexane / IGEPAL CO-520 mixture with a volume ratio of 71 :29. The first emulsion contained a 2.5 M CaCh solution (500 pl) and 1 mg of NAD+. The second emulsion comprised a 25 mM ISfeHPCU solution (500 pl, pH 9) with 3 mg of DOPA. The two emulsions were mixed, stirred for 30 minutes, and then demulsified by adding 50 ml of ethanol to remove the oil phase. The pellet was sequentially washed twice with ethanol and once with 70% ethanol. Subsequently, the particles were redispersed in chloroform containing either DSPE-mPEG2k, and cholesterol (used for ex vivo experiments) or Soy PC at 3 mg and cholesterol at 0.3 mg (used for in vivo experiments). Following chloroform evaporation under a rotary evaporator, the resultant lipid film was rehydrated with 500 pl Tris-HCl buffer (10 mM, pH 7.4) to provide the NAD -NP.

[0092] Biomimetic nanoparticles for in vivo work were prepared by coating platelet membrane on the surface of the NAD+-NP via the extrusion method3,4. In short, platelet membrane vesicles were mixed with NAD+-NP in a 1 : 1 weight ratio of membrane protein to nanoparticles. This mixture was then sequentially extruded through polycarbonate membranes with pore sizes of 400 nm followed by 200 nm using an Avanti mini extruder, maintaining sterile conditions throughout the process. The empty CaP biomimetic NP (P- NP) were prepared following the same procedure by extruding empty CaP NP (without NAD+) with the platelet membrane vesicles. In the biodistribution study, biomimetic nanoparticles were labeled by co-loading 100 pg ATTO-550-labeled tracrRNA and 1 mg NAD+.Example 2 - Preparation of NADH-Lipid-Metal Organic Framework Nanoparticles (NADH-LP-MOF)

[0093] The NADH-LP-MOF NP may be prepared by mixing zinc nitrate, 2- methylimidazole, and NADH in water under ultrasonication to yield the ZIF-8 core, which may be subsequently stabilized by lipid coating via an extrusion process. NADH (6.6 mg) and zinc nitrate hexahydrate (18.6 mg) are dissolved in 35 mL DI water. 2-Methylimidazole (166 mg, dissolved in 10 mL DI water) are added to the NADH solution. The resultant mixture is vortexed for 10 s and is kept still for 5 min to allow MOF growth. The MOF nanoparticles are then pelleted through centrifugation at 10,000 g for 45 min, are redispersed in 1.5 mL water and are ultrasonicated for 30 times. The MOF nanoparticle are mixed with a liposome solution (composed of Soy PC and cholesterol, 10: 1 w / w, 40 mg / mL) and are extruded through a 0.4 pm polycarbonate porous membrane using an Avanti mini extruder to obtain NADH-LP-MOF.Example 3 -Characterization of NAD(H) NP

[0094] NAD+-NP prepared as in Example 1 resulted in nanoparticles with an average hydrodynamic diameter of approximately 165 nm (FIG. 1 A). The morphology of NAD+-NP was determined by TEM (FIG. IB). To achieve the platelet membrane-coated NAD+-NP (i.e., P-NAD+-NP), an extrusion technique was applied. The optimal weight ratio for platelet membrane / NAD+-NP was first determined to be 1 : 1 (FIG. ID). After extrusion, the average hydrodynamic diameter of the nanoparticles increased to about 190 nm, with the increment attributable to the platelet membrane coating (FIG. 1 A). The morphology of P-NAD+NPs were confirmed by TEM, revealing a diameter of about 170 nm (FIG. 1C). The successful application of the platelet membrane coating was further evidenced by the alteration in zeta potential, shifting from a slight negative charge of -5 mV to about -30 mV, consistent with the known surface charge of platelet membrane7(FIG. IE). The loading efficiency of NAD+within the P-NAD+-NP was determined to be 55%, corresponding to a loading content of 5.9%. Stability assessments demonstrated that P-NAD+-NP remained stable for at least one week when stored at 4 °C (FIG. IF).Example 4 - Human Vein Explant Study

[0095] NAD+levels were examined in an ex vivo explant culture using freshly excised, discarded human saphenous veins. Vein explants were exposed to extended storage in saline- supplemented with or without NAD+- NP at a final concentration of mere 10 pM NAD+payload - under ambient environmental conditions for 6 hr, in order to recapitulate the standard workflows of a typical VG bypass procedure. We hypothesize that the nonbuffered saline and room temperature storage could instigate profound bioenergetic and redox crisis in VG, hence rapidly depleting the NAD+pool in EC. Indeed, we were able to observe a significant reduction of NAD+level in tissue homogenates of the ex vivo vein explant culture, in comparison to the fresh vein tissues without exposure to storage in saline or Plasma-Lyte (FIG. 2A). Simultaneously, an increase in inflammatory and thrombogenic phenotypic transcription (P-Selectin) suggesting compromised EC integrity and EC dysfunction for markers P-Selectin and inflammasome genes, including NLRP3 (FIG. 2C) and ILlb (FIG. 2D), as well as a decrease in EC physiological marker gene expression (eNOS) (FIG. 2E) were observed following storage in saline. Conversely, NP-assisted NAD+supplementation in saline effectively rescued the EC dysfunction in ex vivo human vein explants upon extended storage. Taken together, this pilot study shows the utility of NAD+repletion for improved pre-bypass VG storage.Example 5 - NP-assisted Pre-bypass NAD+Repletion Improves Post-bypass VG Patency in a Rat Model In Vivo

[0096] The in vivo efficacy of the NP-assisted NAD+repletion pre-bypass therapy was tested in a rat model of autologous VG bypass. As summarized in FIG. 3 A, following similar ex vivo storage in saline solution - supplemented with or without NAD+-NP, the autologous external jugular veins were subsequently grafted through a cuff-based, end-to- end anastomosis with the common carotid arteries. Due to the arterial hemodynamic injury and the restricted outflow, the VG would undergo constrictive remodeling, leading to IH and ultimately loss of lumen patency. At 2 months post bypass, a significant increase in luminal diameter as well as reduction in IH could be readily observed in VG that were stored in NAD+-CaP NP-supplemented saline prior to bypass (FIG. 3B). Collectively, our current data demonstrated the utility of a pre-bypass ex vivo NAD+-replenishing therapy in mitigating IH in VG after bypass procedure.Example 6 - NAD+-loaded, Biomimetic CaP-NP Differentially Inhibits SMC Proliferation While Preserving EC Health

[0097] To test the role of NAD+biosynthesis (i.e., via the nicotinamide phosphoribosyltransferase (NAMPT)-mediated salvage pathway) in modulating the phenotypic transition of SMC under diseased states, the in vitro efficacy of the platelet membrane-coated NP was tested, with a special emphasis on probing the physiological impact of NAD+repletion on SMC’s hyper-proliferative phenotypic transition. As shown in FIG. 4A, primary culture of human aortic SMC (HASMC) was stimulated with platelet- derived growth factor BB (PDGF-BB), a widely adopted stimulant of SMC proliferation that is abundant upon angioplasty as well as during the pathogenesis of restenosis. Concomitant with PDGF-BB stimulation, SMC was also treated with either PBS, naked NAD+solution, empty biomimetic CaP-NP (P-NP), or NAD+-loaded, biomimetic CaP-NP (P-NAD+-NP) (all at a concentration equivalent to lOpM NAD+payload as determined in pilot studies). In stark contrast to the hyper-proliferative phenotype demonstrated in SMC treated with PDGF-BB, NAD+-replenished SMC displayed a quiescent phenotype following treatment with P-NAD-NP, as evidenced by significantly reduced cell viability and DNA synthesis (BrdU incorporation during the S phase of cell cycle). Naked, non-NP -formulated NAD+solution alone failed to inhibit SMC proliferation, as expected given its inability to bypass plasma membrane. Similarly, empty P-NP was largely ineffective in modulating PDGF-BB-induced SMC hyper-proliferation. A statistically significant, yet marginal benefit was observed with the empty P-NP treatment group in terms of reducing SMC viability.

[0098] Considering the lack of anti-restenotic therapies, we tested whether NAD+-loaded, biomimetic CaP-NP could also promote EC regeneration. As shown in FIG. 4B, primary culture of human aortic EC (HAEC) was challenged with tumor necrosis factor alpha (TNFa) to induce cell death, mimicking the catastrophic cellular events following angioplasty-induced endothelium denudation. Similar to the experimental design in HASMC, TNFa-challenged EC was treated with biomimetic CaP-NP loaded with lOpM NAD+, as well as the appropriate control treatments. TNFa dramatically compromised EC viability and elicited apoptotic activation in HAEC, both of which were successfullyrescued upon treatment with P-NAD+- NP. No protective effect could be observed in any control groups, in which EC was treated with PBS, naked NAD+, or empty P-NP.Example 7 - Targetability, Biodistribution, and Biocompatibility

[0099] To investigate the IH lesion targetability of the platelet membrane-coated biomimetic CaP-NP, we loaded the CaP core with ATTO550-tracrRNA as fluorescent tracer, which closely resembles NAD+in both molecular weight and charge. Immediately following balloon angioplasty in the left common carotid arteries in male SD rats, the fluorescently labelled biomimetic CaP-NP was intravenously injected immediately post the procedure (at 10 mg / kg equivalent to the payload). At 6 hr post injection, the injured carotid arteries, together with the contralateral un-injured carotid arteries as well as major organs, were collected for ex vivo fluorescent IVIS imaging. Consistent with prior efforts utilizing platelet membrane-coated NP of similar sizes but distinct composition, our current study revealed higher enrichment of the biomimetic CaP-NP in injured carotid arteries over uninjured ones (FIG. 5B). Moreover, breaking away from earlier generations of biomimetic nanoplatforms, the platelet membrane-coated CaP-NP displayed greater tropism toward arterial / aortic tissues over other major organs, especially the reticuluendothelial system (liver and spleen), as demonstrated by the significantly greater level of normalized fluorescent signal as shown in FIG. 5C. Overall, in line with our prior studies utilizing the platelet membrane coating for surface functionalization, the IVIS imaging analysis strongly support the IH lesion-targetability and desired biodistribution pattern of the biomimetic CaP-NP.

[0100] Moreover, the is highly biocompatible in rodent models. In male SD rats that received a one-time injection of the biomimetic P-NAD+-NP (equivalent to 10 mg / kg NAD+payload as established in our prior study), we observed no signs of toxicities and tissue structural damages in major organs including liver, spleen, heart, and kidneythereby supporting the feasibility of a more frequent and flexible injection regimen of the NAD+repletion therapy.Example 8 - Targeted NAD+Delivery via the Biomimetic P-NAD+-NP Mitigates Postangioplasty IH In Vivo

[0101] Inspired by our biomimetic P-NAD+-NP’s potent effect in inhibiting SMC hyperproliferation and demonstrated targetability, we set to explore its anti-restenotic efficacy in vivo. Immediately following balloon angioplasty of left common carotid arteries in male SD rats, we provided a single-dose regimen via intravenous injection with the following treatments: saline, empty biomimetic CaP-NP (empty NP), naked NAD+solution (free NAD+), and NAD+-loaded, biomimetic CaP-NP (P-NAD+-NP) — all at a dose equivalent to 10 mg / kg NAD+payload. Rats were euthanized two weeks post angioplasty and aforementioned treatment, followed by histological and morphometric analysis of carotid arteries to determine the lesional extent of IH. As shown in FIG. 6B, accumulation of hyper- proliferative SMC could be observed in the neointimal layer in angioplastied carotid arteries. Further evaluation of IH, as determined by intima-to-media ratio (I / M ratio), revealed the profound anti-restenotic benefits of NAD+-NP in comparison to saline control group in FIG. 6C. As expected, naked, non-formulated NAD+solution displayed no observable effects in altering the IH lesional size. Treatment with the empty biomimetic CaP-NP, however, showed a marginal and statistically non-significant trend toward IH reduction, in accordance with the similar in vitro observation in reducing SMC viability as noted in FIG. 4A. Analysis of the mRNA level of SMC contractility / maturation gene aSMA further corroborated the efficacy of the biomimetic NAD+-NP in mitigating postangioplasty IH (FIG. 6D).Example 9 - Targeted NAD+Delivery via the Biomimetic P-NAD+-CaP NP Accelerates Re-endothelialization Following Angioplasty-induced EC Denudation In Vivo

[0102] Based upon the EC in vitro data and anti-restenotic efficacy data in vivo, we hypothesized that effective NAD+repletion, enabled by our novel biomimetic CaP-NP may hold the key to solve the seemingly irreconcilable dilemma between inhibiting SMC proliferation and promoting EC regeneration. To test this, we utilized the same balloon angioplasty model in rat carotid arteries to induce endothelium denudation, immediately followed by a single-dose regimen of the biomimetic P-NAD+-NP (equivalent to 10 mg / kg NAD+payload) and the aforementioned control treatments. At day 6 post angioplasty, animals were subjected to intravenous injection with Evans Blue dye to help determine the extent of re-endothelialization. As shown in the longitudinally opened carotid arteries inFIG. 7B, the re-endothelialized luminal areas were not permeable to Evans Blue dye, whereas the non-EC-covered areas show prominent staining in blue color. Quantification of the re-endothelialization percentage over baseline level showed significantly improved endothelium regeneration following targeted NAD+repletion with the biomimetic NAD+- NP over control treatment group with saline. Similar to the observations in FIG. 4A and FIG. 6C, a slight yet statistically non-significant increase in re-endothelialization was noted following treatment with the empty biomimetic CaP-NP was observed as shown in FIG. 7B. qPCR analysis of mRNA expression changes for genes representative of EC physiological function / integrity (eNOS), thrombogenicity (tissue factor / TF), impairment of EC healing (CXCL10), and inflammatory cytokine (ICAM1) further validated the EC protection of targeted NAD+repletion in vivo in FIG. 7C. Taken together, the data demonstrate that targeted NAD+delivery, offers a promising solution for safer yet effective management of restenosis.EQUIVALENTS

[0103] While certain embodiments have been illustrated and described, a person with ordinary skill in the art, after reading the foregoing specification, can effect changes, substitutions of equivalents and other types of alterations to the nanoparticles of the present technology or derivatives, prodrugs, or pharmaceutical compositions thereof as set forth herein. Each aspect and embodiment described above can also have included or incorporated therewith such variations or aspects as disclosed in regard to any or all of the other aspects and embodiments.

[0104] The present technology is also not to be limited in terms of the particular aspects described herein, which are intended as single illustrations of individual aspects of the present technology. Many modifications and variations of this present technology can be made without departing from its spirit and scope, as will be apparent to those skilled in the art. Functionally equivalent methods within the scope of the present technology, in addition to those enumerated herein, will be apparent to those skilled in the art from the foregoing descriptions. Such modifications and variations are intended to fall within the scope of the appended claims. It is to be understood that this present technology is not limited to particular methods, conjugates, reagents, compounds, compositions, labeled compounds orbiological systems, which can, of course, vary. All methods described herein can be performed in any suitable order unless otherwise indicated herein or otherwise clearly contradicted by context. It is also to be understood that the terminology used herein is for the purpose of describing particular aspects only and is not intended to be limiting. Thus, it is intended that the specification be considered as exemplary only with the breadth, scope and spirit of the present technology indicated only by the appended claims, definitions therein and any equivalents thereof. No language in the specification should be construed as indicating any non-claimed element as essential.

[0105] The embodiments, illustratively described herein may suitably be practiced in the absence of any element or elements, limitation or limitations, not specifically disclosed herein. Thus, for example, the terms “comprising,” “including,” “containing,” etc. shall be read expansively and without limitation. Additionally, the phrase “consisting essentially of’ will be understood to include those elements specifically recited and those additional elements that do not materially affect the basic and novel characteristics of the claimed technology. The phrase “consisting of’ excludes any element not specified. The terms and expressions employed herein have been used as terms of description and not of limitation, and there is no intention in the use of such terms and expressions of excluding any equivalents of the features shown and described or portions thereof, but it is recognized that various modifications are possible within the scope of the claimed technology. More specifically, it will be understood that each use of terms such as “comprising,” “consisting essentially of,” or “consisting of’, discloses and provides written description and support for the use any of the other terms with the same or any other embodiment described herein.

[0106] In addition, where features or aspects of the disclosure are described in terms of Markush groups, those skilled in the art will recognize that the disclosure is also thereby described in terms of any individual member or subgroup of members of the Markush group. Each of the narrower species and subgeneric groupings falling within the generic disclosure also form part of the technology. This includes the generic description of the technology with a proviso or negative limitation removing any subject matter from the genus, regardless of whether or not the excised material is specifically recited herein.

[0107] As will be understood by one skilled in the art, for any and all purposes, particularly in terms of providing a written description, all ranges disclosed herein also encompass any and all possible subranges and combinations of subranges thereof. Any listed range can be easily recognized as sufficiently describing and enabling the same range being broken down into at least equal halves, thirds, quarters, fifths, tenths, etc. As a nonlimiting example, each range discussed herein can be readily broken down into a lower third, middle third and upper third, etc. As will also be understood by one skilled in the art all language such as “up to,” “at least,” “greater than,” “less than,” and the like, include the number recited and refer to ranges which can be subsequently broken down into subranges as discussed above. Finally, as will be understood by one skilled in the art, a range includes each individual member, and each separate value is incorporated into the specification as if it were individually recited herein.

[0108] All publications, patent applications, issued patents, and other documents (for example, journals, articles and / or textbooks) referred to in this specification are herein incorporated by reference as if each individual publication, patent application, issued patent, or other document was specifically and individually indicated to be incorporated by reference in its entirety. Definitions that are contained in text incorporated by reference are excluded to the extent that they contradict definitions in this disclosure.

[0109] The present technology may include, but is not limited to, the features and combinations of features recited in the following lettered paragraphs, it being understood that the following paragraphs should not be interpreted as limiting the scope of the claims as appended hereto or mandating that all such features must necessarily be included in such claims:A. A method of reducing vein graft injury comprising administering an effective amount of a bioavailable nanoparticle comprising NAD+and / or NADH to the vein to be grafted prior to surgical implantation of the vein graft in a subject.B. The method of Paragraph A, wherein the bioavailable nanoparticle comprises an inorganic core, NAD+and / or NADH, and a coating comprising a lipid bilayer, whereinthe inorganic core is selected from calcium phosphate or a metal organic framework (MOF); the MOF comprises a transition metal ion coordinated to a coordinating ligand, wherein the transition metal ion is selected from the group consisting of zinc, iron, zirconium, copper, and cobalt ions, and the coordinating ligand is selected from an imidazolate ligand or a carboxylate ligand.C. The method of Paragraph A or Paragraph B, wherein administering an effective amount of the bioavailable nanoparticle comprises storing the vein to be grafted in an aqueous solution comprising the effective amount of the bioavailable nanoparticle.D. The method of any one of Paragraphs A-C, wherein the aqueous solution comprises saline and / or one or more additives selected from the group consisting of potassium lactobionate, potassium hydrogen phosphate, magnesium sulfate, raffinose, denosine, glutathione, allopurinol, and hydroxyethyl starch.E. The method of Paragraph C or Paragraph D, wherein the aqueous solution in which the vein to be grafted is stored is at a temperature from 0°C to about 25°C.F. The method of any one of Paragraphs C-E, wherein the aqueous solution in which the vein to be grafted is stored is at a temperature from 0°C to 5°C.G. The method of any one of Paragraphs A-F, wherein the effective amount of NAD+and / or NADH is about 1 pM to about 10 mM.H. The method of any one of Paragraphs A-G, wherein the effective amount of NAD+and / or NADH is about 2 pM to about 100 pM.I. The method of any one of Paragraphs C-H, wherein the effective amount of NAD+and / orNADH is about 5 pM to about 20 pM.J. A method of improving vein graft health comprising administering an effective amount of a bioavailable nanoparticle comprising NAD+and / or NADH and a coating comprising a human cell membrane to a subject post-surgical implantation of the vein graft in the subject, wherein the human cell membrane is selected from thegroup consisting of a human platelet membrane, a human red blood cell membrane, a human white blood cell membrane, mesenchymal stem cell membrane, and a human macrophage cell membrane.K. A method of preventing or treating post-operative restenosis comprising administering an effective amount of a bioavailable nanoparticle comprising NAD+and / or NADH and a coating comprising a human cell membrane to a subject after surgical reconstruction of a blood vessel in the subject, wherein the human cell membrane is selected from the group consisting of a human platelet membrane, a human red blood cell membrane, a human white blood cell membrane, mesenchymal stem cell membrane, and a human macrophage cell membrane.L. The method of Paragraph K, wherein the surgical reconstruction of the blood vessel in the subject comprises balloon angioplasty and / or placement of a stent.M. The method of any one of Paragraphs J-L, wherein the bioavailable nanoparticle comprises an inorganic core and NAD+and / or NADH, and a coating comprising a lipid bilayer, wherein the inorganic core is selected from calcium phosphate or a metal organic framework (MOF); the MOF comprises a transition metal ion coordinated to a coordinating ligand, wherein the transition metal ion is selected from the group consisting of zinc, iron, zirconium, copper, and cobalt ions, and the coordinating ligand is selected from an imidazolate ligand or a carboxylate ligand; and the lipid bilayer comprises or is encapsulated by the human cell membrane.N. The method of any one of Paragraphs J-M, wherein the effective amount of NAD+and / or NADH is 0.1 mg / kg to 200 mg / kg.O. The method of any one of Paragraphs J-N, wherein the effective amount of NAD+and / or NADH is 1 mg / kg to 100 mg / kg.P. The method of any one of Paragraphs J-O, wherein the effective amount of NAD+and / orNADH is 5 mg / kg to 20 mg / kg.Q. The method of any one of Paragraphs J-P, wherein the human cell membrane is the human platelet membrane.R. The method of any one of Paragraphs J-Q, wherein the weight ratio of the human cell membrane coating to the remainder of the bioavailable nanoparticle is 2: 1 to 1 :2.S. The method of any one of Paragraphs J-R, wherein the weight ratio of the human cell membrane coating to the remainder of the bioavailable nanoparticle is about 1 : 1.T. The method of any one of Paragraphs J-S, wherein the bioavailable nanoparticles are administered to the subject by injection.U. The method of any one of Paragraphs A-T, wherein the subject is a human.V. The method of any one of Paragraphs A-U, wherein the bioavailable nanoparticle comprises 1 wt%-50 wt% NAD+or NADH.W. The method of any one of Paragraphs A-V, wherein the bioavailable nanoparticle comprises 1 wt% to 25 wt% NAD+or NADH.X. The method of any one of Paragraphs A-W, wherein the bioavailable nanoparticle comprises 10 wt%-75 wt% lipid bilayer.Y. The method of any one of Paragraphs A-W, wherein the bioavailable nanoparticle comprises 30 wt%-75 wt% human cell membrane.Z. The method of any one of Paragraphs B-I or M-Y, wherein the lipid bilayer comprises lipids selected from the group consisting of of L-a-phosphatidylcholine (PC), 1,2- dioleoyl-sn-glycero-3 -phosphate (DOPA), l,2-dioleoyl-3 -trimethylammonium - propane (DOTAP), l,2-dioleoyl-sn-glycero-3 -phosphocholine (DOPC), 1,2- dioleoyl-sn-glycero-3 -phosphoethanolamine (DOPE), l,2-dioleoyl-sn-glycero-3- phospho-L-serine (DOPS), l,2-distearoyl-sn-glycero-3-phosphoethanolamine-N- [methoxy(polyethylene glycol)] (DSPE-PEG), cholesterol, a cell membrane extracted from a red blood cell, macrophage, neutrophil or platelet, and combinations of two or more thereof.AA. The method of any one of Paragraphs B-I or M-Z, wherein the lipid bilayer comprises lipids conjugated to polyethylene glycol) (PEG).AB. The method of any one of Paragraphs B-I or M-AA, wherein up to 100 mol% of the lipids in the lipid bilayer are conjugated to PEG.AC. The method of any one of Paragraphs B-I or M-AB, wherein the lipids of the lipid bilayer comprise a combination of PC and DOPA and / or cholesterol.AD. The method of any one of Paragraphs B-I or M-AC, wherein the lipid bilayer comprises DSPE-PEG wherein: the PEG has a free terminus selected from the group consisting of OH, O-C1-4 alkyl ether, NH2, NHR, COOH, COOR, wherein R is an alkyl or alkenyl group, a dye, a targeting ligand, and a metal chelating ligand, and the PEG has a number average molecular weight ranging from 300 to 10,000 Da.AE. The method of any one of Paragraphs B-AD, comprising 10 - 90 wt% inorganic core.AF. The method of any one of Paragraphs B-AE, wherein the inorganic core is calcium phosphate.AG. The method of any one of Paragraphs B-AF, wherein the inorganic core is MOF.AH. The method of any one of Paragraphs B-AG, where the coordinating ligand is selected from the group consisting of imidazole, 2-methyl-imidazole, benzimidazole, 5- methylbenzimidazole, terephthalic acid, 2-methyl-pterphthalic acid, 2-hydroxy- terphthalic acid, and 2-amino-terphthalic acid, benzene-l,3,5-tricarboxylic acid, l,3,5-tris(4-carboxyphenyl)benzene, 2,6-naphthalenedicarboxylic acid, 4,4',4"-s- triazine-2,4,6-triyl-tribenzoic acid, and 2,5-dihydroxyterephthalic acid.Al. The method of any one of Paragraphs B-AH, wherein the MOF comprises zinc ions and imidazolate ligands.AJ. The method of any one of Paragraphs B-AI, wherein the imidazolate ligand is selected from imidazole, 2-methyl-imidazole, benzimidazole, or 5 -methylbenzimidazole.AK. The method of any one of Paragraphs B-AJ, wherein the imidiazolate ligand is selected from 2-methyl-imidazole.AL. The method of any one of Paragraphs A-AK, wherein the bioavailable nanoparticle has an average hydrodynamic diameter of from 60 to less than 1000 nm.AM. The method of any one of Paragraphs A-AL, wherein bioavailable nanoparticles are administered as a pharmaceutical composition comprising a pharmaceutically acceptable carrier.AN. A bioavailable nanoparticle comprising NAD+and / or NADH and a coating comprising a human cell membrane selected from the group consisting of a human platelet membrane, a human red blood cell membrane, a human white blood cell membrane, mesenchymal stem cell membrane, and a human macrophage cell membrane.AO. The bioavailable nanoparticle of Paragraph AN, further comprising an inorganic core and NAD+and / or NADH, and a coating comprising a lipid bilayer, wherein the inorganic core is selected from calcium phosphate or a metal organic framework (MOF); the MOF comprises a transition metal ion coordinated to a coordinating ligand, wherein the transition metal ion is selected from the group consisting of zinc, iron, zirconium, copper, and cobalt ions, and the coordinating ligand is selected from an imidazolate ligand or a carboxylate ligand; and the lipid bilayer comprises or is encapsulated by the human cell membrane.AP. The bioavailable nanoparticle of Paragraph AN or Paragraph AO, wherein the human cell membrane is the human platelet membrane.AQ. The bioavailable nanoparticle of any one of Paragraphs AN-AP, wherein the weight ratio of the human cell membrane coating to the remainder of the bioavailable nanoparticle is 2: 1 to 1 :2.AR. The bioavailable nanoparticle of any one of Paragraphs AN-AQ, wherein the weight ratio of the human cell membrane coating to the remainder of the bioavailable nanoparticle is about 1 : 1.AS. The bioavailable nanoparticle of any one of Paragraphs AN-AR, wherein the bioavailable nanoparticle comprises 1 wt%-50 wt% NAD+or NADH.AT. The bioavailable nanoparticle of any one of Paragraphs AN-AS, wherein the bioavailable nanoparticle comprises 1 wt% to 25 wt% NAD+or NADH.AU. The bioavailable nanoparticle of any one of Paragraphs AN-AT, wherein the bioavailable nanoparticle comprises 10 wt%-75 wt% lipid bilayer.AV. The bioavailable nanoparticle of any one of Paragraphs AN-AU, wherein the bioavailable nanoparticle comprises 30 wt%-75 wt% human cell membrane.AW. The bioavailable nanoparticle of any one of Paragraphs AO- AV, wherein the lipid bilayer comprises lipids selected from the group consisting of of L-a- phosphatidylcholine (PC), l,2-dioleoyl-sn-glycero-3 -phosphate (DOPA), 1,2- dioleoyl-3-trimethylammonium-propane (DOTAP), l,2-dioleoyl-sn-glycero-3- phosphocholine (DOPC), l,2-dioleoyl-sn-glycero-3 -phosphoethanolamine (DOPE), l,2-dioleoyl-sn-glycero-3-phospho-L-serine (DOPS), l,2-distearoyl-sn-glycero-3- phosphoethanolamine-N-[methoxy(polyethylene glycol)] (DSPE-PEG), cholesterol, a cell membrane extracted from a red blood cell, macrophage, neutrophil or platelet, and combinations of two or more thereof.AX. The bioavailable nanoparticle of any one of Paragraphs AO-AW, wherein the lipid bilayer comprises lipids conjugated to polyethylene glycol) (PEG).AY. The bioavailable nanoparticle of any one of Paragraphs AO-AX, wherein up to 100 mol% of the lipids in the lipid bilayer are conjugated to PEG.AZ. The bioavailable nanoparticle of any one of Paragraphs AO-AY, wherein the lipids of the lipid bilayer comprise a combination of PC and DOPA and / or cholesterol.BA. The bioavailable nanoparticle of any one of Paragraphs AO-AZ, wherein the lipid bilayer comprises DSPE-PEG wherein: the PEG has a free terminus selected from the group consisting of OH, O-C1-4 alkyl ether, NH2, NHR, COOH, COOR, wherein R is an alkyl or alkenyl group, a dye, a targeting ligand, and a metal chelating ligand, and the PEG has a number average molecular weight ranging from 300 to 10000 Da.BB. The bioavailable nanoparticle of any one of Paragraphs AO-BA comprising 10 - 90 wt% inorganic core.BC. The bioavailable nanoparticle of any one of Paragraphs AO-BB, wherein the inorganic core is calcium phosphate.BD. The bioavailable nanoparticle of any one of Paragraphs AO-BC, wherein the inorganic core is MOF.BE. The bioavailable nanoparticle of Paragraph BD, where the coordinating ligand is selected from the group consisting of imidazole, 2-methyl-imidazole, benzimidazole, 5-methylbenzimidazole, terephthalic acid, 2-methyl-pterphthalic acid, 2-hydroxy- terphthalic acid, and 2-amino-terphthalic acid, benzene-l,3,5-tricarboxylic acid, l,3,5-tris(4-carboxyphenyl)benzene, 2,6-naphthalenedicarboxylic acid, 4,4',4"-s- triazine-2,4,6-triyl-tribenzoic acid, and 2,5-dihydroxyterephthalic acid.BF. The bioavailable nanoparticle of Paragraph BD or Paragraph BE, wherein the MOF comprises zinc ions and imidazolate ligands.BG. The bioavailable nanoparticle of any one of Paragraphs BD-BF, wherein the imidazolate ligand is selected from imidazole, 2-methyl-imidazole, benzimidazole, or 5-methylbenzimidazole.BH. The bioavailable nanoparticle of any one of Paragraphs BD-BG, wherein the imidiazolate ligand is selected from 2-methyl-imidazole.BI. The bioavailable nanoparticle of any one of Paragraphs BD-BH, wherein the bioavailable nanoparticle has an average hydrodynamic diameter of from 60 to less than 1000 nm.

[0110] Other embodiments are set forth in the following claims, along with the full scope of equivalents to which such claims are entitled.

Claims

WHAT IS CLAIMED IS:

1. A method of reducing vein graft injury comprising administering an effective amount of a bioavailable nanoparticle comprising NAD+and / or NADH to the vein to be grafted prior to surgical implantation of the vein graft in a subject.

2. The method of claim 1, wherein the bioavailable nanoparticle comprises an inorganic core, NAD+and / or NADH, and a coating comprising a lipid bilayer, wherein the inorganic core is selected from calcium phosphate or a metal organic framework (MOF); the MOF comprises a transition metal ion coordinated to a coordinating ligand, wherein the transition metal ion is selected from the group consisting of zinc, iron, zirconium, copper, and cobalt ions, and the coordinating ligand is selected from an imidazolate ligand or a carboxylate ligand.

3. The method of claim 1, wherein administering an effective amount of the bioavailable nanoparticle comprises storing the vein to be grafted in an aqueous solution comprising the effective amount of the bioavailable nanoparticle.

4. The method of claim 1, wherein the aqueous solution comprises saline and / or one or more additives selected from the group consisting of potassium lactobionate, potassium hydrogen phosphate, magnesium sulfate, raffinose, denosine, glutathione, allopurinol, and hydroxy ethyl starch.

5. The method of claim 3, wherein the aqueous solution in which the vein to be grafted is stored is at a temperature from 0°C to about 25°C.

6. The method of claim 3, wherein the aqueous solution in which the vein to be grafted is stored is at a temperature from 0°C to 5°C.

7. The method of claim 1, wherein the effective amount of NAD+and / or NADH is about 1 pM to about 10 mM.

8. The method of claim 1, wherein the effective amount of NAD+and / or NADH is about 2 pM to about 100 pM.

9. The method of any one of claims 1-8, wherein the effective amount of NAD+and / or NADH is about 5 pM to about 20 pM.

10. A method of improving vein graft health comprising administering an effective amount of a bioavailable nanoparticle comprising NAD+and / or NADH and a coating comprising a human cell membrane to a subject post-surgical implantation of the vein graft in the subject, wherein the human cell membrane is selected from the group consisting of a human platelet membrane, a human red blood cell membrane, a human white blood cell membrane, mesenchymal stem cell membrane, and a human macrophage cell membrane.

11. A method of preventing or treating post-operative restenosis comprising administering an effective amount of a bioavailable nanoparticle comprising NAD+and / or NADH and a coating comprising a human cell membrane to a subject after surgical reconstruction of a blood vessel in the subject, wherein the human cell membrane is selected from the group consisting of a human platelet membrane, a human red blood cell membrane, a human white blood cell membrane, mesenchymal stem cell membrane, and a human macrophage cell membrane.

12. The method of claim 11, wherein the surgical reconstruction of the blood vessel in the subject comprises balloon angioplasty and / or placement of a stent.

13. The method of claim 10 or claim 11, wherein the bioavailable nanoparticle comprises an inorganic core and NAD+and / or NADH, and a coating comprising a lipid bilayer, wherein the inorganic core is selected from calcium phosphate or a metal organic framework (MOF); the MOF comprises a transition metal ion coordinated to a coordinating ligand, wherein the transition metal ion is selected from the group consisting of zinc, iron, zirconium, copper, and cobalt ions, and thecoordinating ligand is selected from an imidazolate ligand or a carboxylate ligand; and the lipid bilayer comprises or is encapsulated by the human cell membrane.

14. The method of claim 10 or claim 11, wherein the effective amount of NAD+and / or NADH is 0.1 mg / kg to 200 mg / kg.

15. The method of claim 10 or claim 11, wherein the effective amount of NAD+and / or NADH is 1 mg / kg to 100 mg / kg.

16. The method of claim 10 or claim 11, wherein the effective amount of NAD+and / or NADH is 5 mg / kg to 20 mg / kg.

17. The method of claim 10 or claim 11, wherein the human cell membrane is the human platelet membrane.

18. The method of claim 10 or claim 11, wherein the weight ratio of the human cell membrane coating to the remainder of the bioavailable nanoparticle is 2: 1 to 1 :2.

19. The method of claim 10 or claim 11, wherein the weight ratio of the human cell membrane coating to the remainder of the bioavailable nanoparticle is about 1 : 1.

20. The method of claim 10 or claim 11, wherein the bioavailable nanoparticles are administered to the subject by injection.

21. The method of any one of claims 1, 10, or 11, wherein the subject is a human.

22. The method of any one of claims 1, 10, or 11, wherein the bioavailable nanoparticle comprises 1 wt%-50 wt% NAD+or NADH.

23. The method of any one of claims 1, 10, or 11, wherein the bioavailable nanoparticle comprises 1 wt% to 25 wt% NAD+or NADH.

24. The method of any one of claims 1, 10, or 11, wherein the bioavailable nanoparticle comprises 10 wt%-75 wt% lipid bilayer.

25. The method of any one of claims 1, 10, or 11, wherein the bioavailable nanoparticle comprises 30 wt%-75 wt% human cell membrane.

26. The method of claim 3, wherein the lipid bilayer comprises lipids selected from the group consisting of of L-a-phosphatidylcholine (PC), l,2-dioleoyl-sn-glycero-3- phosphate (DOPA), l,2-dioleoyl-3-trimethylammonium-propane (DOTAP), 1,2- dioleoyl-sn-glycero-3 -phosphocholine (DOPC), l,2-dioleoyl-sn-glycero-3- phosphoethanolamine (DOPE), 1 ,2-dioleoyl-sn-glycero-3 -phospho-L-serine (DOPS), l,2-distearoyl-sn-glycero-3-phosphoethanolamine-N- [methoxy(polyethylene glycol)] (DSPE-PEG), cholesterol, a cell membrane extracted from a red blood cell, macrophage, neutrophil or platelet, and combinations of two or more thereof.

27. The method of claim 2, wherein the lipid bilayer comprises lipids conjugated to polyethylene glycol) (PEG).

28. The method of claim 2, wherein up to 100 mol% of the lipids in the lipid bilayer are conjugated to PEG.

29. The method of claim 2, wherein the lipids of the lipid bilayer comprise a combination of PC and DOPA and / or cholesterol.

30. The method of claim 2, wherein the lipid bilayer comprises DSPE-PEG wherein: the PEG has a free terminus selected from the group consisting of OH, O-Ci- 4 alkyl ether, NH2, NHR, COOH, COOR, wherein R is an alkyl or alkenyl group, a dye, a targeting ligand, and a metal chelating ligand, and the PEG has a number average molecular weight ranging from 300 to 10,000 Da.

31. The method of claim 2, comprising 10 - 90 wt% inorganic core.

32. The method of claim 2, wherein the inorganic core is calcium phosphate.

33. The method of claim 2, wherein the inorganic core is MOF.

34. The method of claim 2, where the coordinating ligand is selected from the group consisting of imidazole, 2-methyl-imidazole, benzimidazole, 5- methylbenzimidazole, terephthalic acid, 2-methyl-pterphthalic acid, 2-hydroxy- terphthalic acid, and 2-amino-terphthalic acid, benzene-l,3,5-tricarboxylic acid, l,3,5-tris(4-carboxyphenyl)benzene, 2,6-naphthalenedicarboxylic acid, 4,4',4"-s- triazine-2,4,6-triyl-tribenzoic acid, and 2,5-dihydroxyterephthalic acid.

35. The method of claim 2, wherein the MOF comprises zinc ions and imidazolate ligands.

36. The method of claim 2, wherein the imidazolate ligand is selected from imidazole, 2-methyl-imidazole, benzimidazole, or 5 -methylbenzimidazole.

37. The method of claim 2, wherein the imidiazolate ligand is selected from 2-methyl- imidazole.

38. The method of any one of claims 1, 10, or 11, wherein the bioavailable nanoparticle has an average hydrodynamic diameter of from 60 to less than 1000 nm.

39. The method of any one of claims 1, 10, or 11, wherein bioavailable nanoparticles are administered as a pharmaceutical composition comprising a pharmaceutically acceptable carrier.

40. A bioavailable nanoparticle comprising NAD+and / or NADH and a coating comprising a human cell membrane selected from the group consisting of a human platelet membrane, a human red blood cell membrane, a human white blood cell membrane, mesenchymal stem cell membrane, and a human macrophage cell membrane.

41. The bioavailable nanoparticle of claim 40, further comprising an inorganic core and NAD+and / or NADH, and a coating comprising a lipid bilayer, wherein the inorganic core is selected from calcium phosphate or a metal organic framework (MOF);the MOF comprises a transition metal ion coordinated to a coordinating ligand, wherein the transition metal ion is selected from the group consisting of zinc, iron, zirconium, copper, and cobalt ions, and the coordinating ligand is selected from an imidazolate ligand or a carboxylate ligand; and the lipid bilayer comprises or is encapsulated by the human cell membrane.

42. The bioavailable nanoparticle of claim 40, wherein the human cell membrane is the human platelet membrane.

43. The bioavailable nanoparticle of claim 40, wherein the weight ratio of the human cell membrane coating to the remainder of the bioavailable nanoparticle is 2: 1 to 1 :2.

44. The bioavailable nanoparticle of claim 40, wherein the weight ratio of the human cell membrane coating to the remainder of the bioavailable nanoparticle is about 1 : 1.

45. The bioavailable nanoparticle of claim 40, wherein the bioavailable nanoparticle comprises 1 wt%-50 wt% NAD+or NADH.

46. The bioavailable nanoparticle of claim 40, wherein the bioavailable nanoparticle comprises 1 wt% to 25 wt% NAD+or NADH.

47. The bioavailable nanoparticle of claim 40, wherein the bioavailable nanoparticle comprises 10 wt%-75 wt% lipid bilayer.

48. The bioavailable nanoparticle of claim 40, wherein the bioavailable nanoparticle comprises 30 wt%-75 wt% human cell membrane.

49. The bioavailable nanoparticle of claim 41, wherein the lipid bilayer comprises lipids selected from the group consisting of of L-a-phosphatidylcholine (PC), 1,2-dioleoyl- sn-glycero-3 -phosphate (DOPA), l,2-dioleoyl-3 -trimethylammonium -propane (DOTAP), l,2-dioleoyl-sn-glycero-3 -phosphocholine (DOPC), 1,2-dioleoyl-sn- glycero-3 -phosphoethanolamine (DOPE), l,2-dioleoyl-sn-glycero-3-phospho-L- serine (DOPS), l,2-distearoyl-sn-glycero-3-phosphoethanolamine-N-[methoxy(polyethylene glycol)] (DSPE-PEG), cholesterol, a cell membrane extracted from a red blood cell, macrophage, neutrophil or platelet, and combinations of two or more thereof.

50. The bioavailable nanoparticle of claim 41, wherein the lipid bilayer comprises lipids conjugated to polyethylene glycol) (PEG).

51. The bioavailable nanoparticle of claim 41, wherein up to 100 mol% of the lipids in the lipid bilayer are conjugated to PEG.

52. The bioavailable nanoparticle of claim 41, wherein the lipids of the lipid bilayer comprise a combination of PC and DOPA and / or cholesterol.

53. The bioavailable nanoparticle of claim 41, wherein the lipid bilayer comprises DSPE-PEG wherein: the PEG has a free terminus selected from the group consisting of OH, O-Ci- 4 alkyl ether, NH2, NHR, COOH, COOR, wherein R is an alkyl or alkenyl group, a dye, a targeting ligand, and a metal chelating ligand, and the PEG has a number average molecular weight ranging from 300 to 10000 Da.

54. The bioavailable nanoparticle of claim 41 comprising 10 - 90 wt% inorganic core.

55. The bioavailable nanoparticle of claim 41, wherein the inorganic core is calcium phosphate.

56. The bioavailable nanoparticle of claim 41, wherein the inorganic core is MOF.

57. The bioavailable nanoparticle of claim 56, where the coordinating ligand is selected from the group consisting of imidazole, 2-methyl-imidazole, benzimidazole, 5- methylbenzimidazole, terephthalic acid, 2-methyl-pterphthalic acid, 2-hydroxy- terphthalic acid, and 2-amino-terphthalic acid, benzene-l,3,5-tricarboxylic acid, l,3,5-tris(4-carboxyphenyl)benzene, 2,6-naphthalenedicarboxylic acid, 4,4',4"-s- triazine-2,4,6-triyl-tribenzoic acid, and 2,5-dihydroxyterephthalic acid.

58. The bioavailable nanoparticle of claim 57, wherein the MOF comprises zinc ions and imidazolate ligands.

59. The bioavailable nanoparticle of claim 58, wherein the imidazolate ligand is selected from imidazole, 2-methyl-imidazole, benzimidazole, or 5 -methylbenzimidazole.

60. The bioavailable nanoparticle of claim 59, wherein the imidiazolate ligand is 2- methyl-imidazole.

61. The bioavailable nanoparticle of claim 56, wherein the bioavailable nanoparticle has an average hydrodynamic diameter of from 60 to less than 1000 nm.

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