TECHNICAL FIELD

Disclosed herein are formulations for sustained transdermal delivery of active agents that enhance levels of nitric oxide in the vasculature and/or limit the production of reactive oxygen species, thus providing new methods of both preventing and treating endothelial dysfunction and its many clinical consequences.

BACKGROUND

Delivery of many therapeutic agents remains a challenge due to bioavailability issues that preclude achieving therapeutic levels either systemically or locally at a targeted site. Poor bioavailability arises from various factors including low solubility, low uptake from the gut, and rapid elimination from the circulation due to rapid breakdown by the liver (first pass limitations). As a result, many promising therapeutics with the appropriate activity cannot be directly delivered in an effective manner to afflicted sites such as the lungs, arthritic joints, inner ear, sinuses etc. Meanwhile, oral or IV routes of administration often do not provide adequate therapeutic levels at the targeted local site.

Nitric oxide (NO) has known systemic benefits including the capability of reversing inflammation, preventing and reversing endothelial dysfunction, repolarizing activated macrophages, deactivating activated platelets, protecting/restoring the endothelial lining of blood vessels (glycocalyx) and antimicrobial/antiviral activity. Proinflammatory insults result in enhanced production of reactive oxygen species (ROS) which drives the development and continuation of endothelial dysfunction. The enhanced levels of ROS set in motion multiple events that cause dysregulation of the vasculature. The ROS degrades the glycocalyx which results in loss of vascular integrity, enhanced cells (monocytes, red blood cells, neutrophils and platelets) access to and adhesion to the endothelial surface, loss of sheer stress mediated NO production and loss of superoxide dismutase which limits ROS. Enhanced ROS causes endothelial nitric oxide synthase (eNOS) to stop production of NO in endothelium and actually start generating more ROS. Enhancing NO levels either with direct supplementation or enhanced production can dramatically reverse this cycle and restore vascular homeostasis. However, systemic delivery of NO is a challenge due to the short lifetime of the NO molecule and limited access to the circulation. A need exists for delivery system that provides efficient and effective enhancement of NO levels in the vasculature and/or the intrinsic regulation of NO levels and bioactivity.

SUMMARY

The delivery system and methods disclosed herein address such a need. The strategy presented in this patent document both enhances NO levels/production directly and reduces ROS production which also results in restoration of NO levels. NO is essential for normal vascular health and function. The presented strategy also restores the integrity of the glycocalyx which allows for the resumption of flow mediated regulation of NO activity and the preservation of vascular integrity. Through these mechanisms, the presented formulations/strategy can be used to assist in preventing or delaying the development of endothelial dysfunction, limit or reverse endothelial dysfunction, and prevent the development of the many clinical consequences of untreated endothelial dysfunction.

The agent delivered via the formulation disclosed herein is able to enhance systemic nitric oxide levels in the endothelium, either by boosting the NO production or by releasing NO into the circulation. By upregulating the systemic NO level or delivering an effective amount of NO to the circulation, the formulation and kit disclosed herein can be applied to the treatment of various conditions or diseases. Examples include transdermal treatment of acute and chronic inflammatory conditions, transdermal prevention and reversal of endothelial dysfunction, risk reduction of cytokine storm or cytokine storm phenomena (release of an abnormal level or a higher than normal level of pro-inflammatory cytokines) in patients with conditions that create underlying endothelial dysfunction, for example COVID-19 or SARS-CoV-2 infection, topical treatment of dermatological conditions, anti-aging skin treatments, ophthalmological conditions, aerosol based treatments of pulmonary conditions, topical treatments of infections, treatment of red blood cells to improve storage properties and reverse storage lesions thus improving safety and efficacy of stored red blood cells, loading red blood cells with therapeutic agents prior to transfusion, stabilizing red blood cells via intravenous (IV) delivery of RBC stabilizing agents, IV interventions to treat cytokine storm phenomena and related acute inflammatory crises, aerosol treatment and prevention of acute respiratory distress syndrome (ARDS) and other conditions that destroy lung tissue through excessive oxidative damage and the ensuing inflammation, and loading medical sponges with therapeutic agents for use, for example in the ear, nose, mouth, rectum, and vagina. Sustained transdermal systemic delivery can also be achieved to harness the therapeutic potential of NO for various chronic diseases and conditions.

An aspect of the present disclosure provides a transdermal formulation for enhancing systemic nitric oxide (NO), comprising:

(a) an effective amount of an NO booster and/or an NO precursor, wherein the NO booster increases systemic production of NO, and wherein the NO precursor comprises an NO releasing agent or derives or produces a NO releasing agent;

(b) a solvent in an amount sufficient to dissolve the effective amount of the NO booster or the NO precursor; and optionally

(c) a fatty acid,

wherein upon administration the effective amount of the NO booster and/or the NO released from the releasing agent is delivered transdermally.

NO boosters useful in the formulation of the present invention include polyphenols, flavonoids, stilbenoid, secosteroid and other phytochemicals and natural products which promote formation of NO. In some embodiments, the NO booster comprises at least one of an agent selected from the group consisting of polyphenol, flavonoid, stilbenoid, secosteroid, and natural products of the like. In some embodiments, the NO booster includes at least one of curcumin, demethoxycurcumin, bisdemethoxycurcumin, quercetin, berberine, resveratrol and vitamin D. In some embodiments, the NO booster includes quercetin and at least one curcuminoid (e.g. curcumin).

NO precursors useful in the formulation comprises a S-nitrosothiol-containing molecule, or a thiol-containing molecule and a nitrite source. In some embodiments, the formulation includes a S-nitrosothiol-containing molecule as a NO precursor. In some embodiments, the formulation includes, as the NO precursor, a thiol-containing molecule, a nitrite source (e.g. nitrite loaded nanoparticles), and optionally an acid source physically separate from the nitrite source, wherein contact of the acid source with the nitrite source nitrosates a thiol group of the thiol-containing molecule. In some embodiments, the nitrite source and the acid source are separately enclosed in permeable or frangible pouches.

In some embodiments, the polyol is polyethylene glycol having a molecular weight ranging from 100 to about 1000. In some embodiments, the fatty acid is myristic acid.

In some embodiments, the formulation provides continued release of the NO booster or the NO releasing agent from the NO precursor (e.g. S-nitrosothiol-containing molecule) over a period of about about 8, about 10, about 15, about 20, about 24 or about 48 hours.

In some embodiments, the formulation further includes a thickener which keeps the transdermal formulation in a semi-solid or solid form. In some embodiments, the thickener is petroleum jelly, cocoa butter or polyalkylene glycol of MW of more than 2k Da.

Another aspect of this invention provides a kit or transdermal delivery system, incorporating the transdermal formulation disclosed herein. In some embodiments, the transdermal delivery system is a kit or nebulizer.

A further aspect provides a method of increasing systemic NO level in a subject. The method includes administering to the subject the transdermal formulation disclosed herein.

A further aspect provides a method of treating a disease or condition by administering to the subject the transdermal formulation disclosed herein. In some embodiments, the disease or condition is associated with endothelial dysfunction.

A further aspect provides a method of incorporating an agent into a cell by contacting the cell with the formulation disclosed herein.

