Transdermal delivery formulations

Abstract

To provide a transdermal formulation for the delivery of one or more active agents to a subject.SOLUTION: A transdermal formulation comprises: (a) an effective amount of a NO booster comprising one or both of an effective amount of a curcuminoid and an effective amount of at least one flavonoid; (b) a polyol in an amount sufficient to dissolve the effective amount of the curcuminoid and the at least one flavonoid; and optionally (c) fatty acid, wherein the polyol and the fatty acid are in a ratio ranging from about 10:1 to about 50:1 by weight, wherein the amounts of the polyol and the fatty acid are selected so that upon administration the effective amount of the NO booster is delivered transdermally. The formulation can be applied to the treatment of various diseases or conditions associated with an elevated CD38 level.SELECTED DRAWING: None

Description

[Technical field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application is a continuation-in-part of U.S. Application No. 18 / 051,266, filed October 31, 2022, which is a divisional application of U.S. Patent No. 11,484,493, which is a continuation of International Application No. PCT / US2021 / 058611, filed November 9, 2021, claiming benefit of priority to Provisional Application No. 63 / 111,160, filed November 9, 2020, No. 63 / 161,696, filed March 16, 2021, and No. 63 / 235,880, filed August 23, 2021, the disclosures of all of which are incorporated by reference in their entireties herein.

[0002] Technical Field Disclosed herein is a formulation for sustained transdermal delivery of active agents that results in a reduction in CD38 ectoenzyme activity, such as NADase activity; thereby enhancing NAD+ levels and promoting the activity of NAD+-dependent sirtuins. In so doing, this approach provides both a new method of prevention and treatment of various diseases or conditions associated with elevated levels of CD38. [Background technology]

[0003] background Delivery of many therapeutic agents remains a challenge due to bioavailability issues that prevent achieving therapeutic levels systemically or locally at the target site. Poor bioavailability results from a variety of factors, including low solubility, low uptake from the intestine, and rapid elimination from the circulation due to rapid degradation by the liver (first-pass limitation). As a result, many promising therapeutic agents with adequate activity cannot be delivered in an effective manner directly to affected sites such as the lungs, arthritic joints, inner ear, and sinuses. Meanwhile, oral or IV administration routes often do not provide adequate therapeutic levels at the target local site.

[0004] Nitric oxide (NO) has known systemic benefits, including the ability to reverse inflammation, prevent and reverse endothelial dysfunction, repolarize activated macrophages, inactivate activated platelets, protect / restore the endothelial lining of blood vessels (glycocalyx), and antibacterial / antiviral activity. Inflammation-induced injury enhances the production of reactive oxygen species (ROS) that drive the onset and continuation of endothelial dysfunction. Enhanced levels of ROS initiate multiple events that cause dysregulation of the vasculature. ROS degrade the glycocalyx, which results in loss of vascular integrity, enhanced access and adhesion of circulating cells (monocytes, red blood cells, neutrophils, and platelets) to the endothelial surface, loss of shear stress-mediated NO production, and loss of superoxide dismutase, which limits ROS. When ROS are enhanced, endothelial nitric oxide synthase (eNOS) stops producing NO in the endothelium and more ROS actually begins to be generated. Increasing NO levels, either by 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 its limited access to the circulation.

[0005] CD38 plays a central role in multiple organ systems and tissues. CD38 is the key enzyme for nicotinamide adenine dinucleotide (NAD) degradation in mammalian cells. Reduced NAD levels are closely related to metabolic syndrome and age-related diseases. In the vasculature, in terms of treating endothelial dysfunction, lowering CD38 levels greatly alleviated angiotensin II (Ang II)-induced vascular remodeling in mice, as indicated by lowered blood pressure; reduced vascular media thickness, media-to-lumen ratio, and collagen deposition; and restored elastin expression. Lowering CD38 levels significantly attenuated Ang II-induced vascular senescence by suppressing the biogenesis, secretion, and internalization of senescence-associated small extracellular vesicles (SA-sEVs), which promote senescence of adjacent non-injured VSMCs. Furthermore, the protective effect of CD38 deficiency on VSMC aging was related to the restoration of lysosomal dysfunction, especially with regard to the maintenance of sirtuin-mediated mitochondrial homeostasis and activation of the mitochondria-lysosomal axis in VSMCs. Thus, CD38 activity, and the resulting associated intracellular NAD reduction, are essential for Ang II-induced VSMC (vascular smooth muscle cell) aging and vascular remodeling, which may result in cardiovascular disease and renal failure. Similar results have been reported for the centrality of CD38 / NAD+ activity in other disease states, including osteoarthritis, lupus, neuropathy, and age-related physical and cognitive decline. The central role of CD38 arises in part due to any of a number of inflammatory triggers, including the accumulation of senescent cells, which gives rise to its overexpression in activated immune cells, such as macrophages and microglial cells, and on the activated intima of all blood vessels. [Prior art documents] [Patent documents]

[0006] [Patent Document 1] US 2014 / 0199391 A1 [Patent Document 2] U.S. 2016 / 0374960 A1 [Patent Document 3] U.S. 2019 / 0201478 A1 [Patent Document 4] WO 2010 / 123547 A1 [Patent Document 5] JP 2014504592 A [Patent Document 6] JP 2008520694 A [Non-Patent Document]

[0007] [Non-Patent Document 1] Boscariol et al., Transdermal permeation of curcumin promoted by choline geranate ionic liquid: Potential for the treatment of skin diseases, Saudi Pharmaceutical Journal 30 (2022) 382-397 [Summary of the Invention]

[0008] Summary This patent document provides a transdermal formulation for delivery of one or more active agents to a subject in the treatment of diseases or conditions associated with elevated CD38 levels, which in turn cause a decrease in NAD+ levels. Low NAD+ promotes mitochondrial dysfunction and limits the activity of sirtuins. Sirtuin activity is necessary to maintain NO production in the endothelium and shut down oxidative and nitrosative stress brought about by activated immune cells, such as the M1 population of macrophages and microglial cells. CD38 is expressed on the surface of macrophages, microglial cells and other immunoactive white blood cells, as well as on the intima of blood vessels. Increased levels of CD38 are caused by diseases or conditions, such as trauma, inflammation, infection, radiation, chemotherapy, and excessive amounts of senescent cells. Enhanced levels of CD38 then promote further cellular senescence, inflammation and oxidative stress, which further stimulates more CD38 production, creating a cycle of persistent inflammation that is difficult to break.

[0009] The transdermal formulation disclosed herein results in a reduction in CD38 ectoenzyme activity, such as NADase activity; thereby enhancing NAD+ levels and promoting the activity of NAD+-dependent sirtuins.In so doing, this approach provides both a new method of prevention and treatment for a variety of diseases, including those resulting from endothelial dysfunction following acute or chronic inflammatory injury, age-related physical and cognitive skill decline, age-related cardiac tissue changes, vascular hypertrophy, osteoarthritis, peripheral neuropathy and long-COVID.Furthermore, this approach overcomes the negative consequences of CD38 overproduction on the ability of stem cells to differentiate into mature cells, which is at least partially responsible for chronic inflammatory anemia and the ineffectiveness of intrinsic stem cells and stem cell therapy to repair damaged tissues.

[0010] The formulations disclosed herein for local delivery provide direct delivery to both plasma and blood cells, thus avoiding first-pass loss by the gastrointestinal tract and liver. In addition, red blood cells loaded with active agents can function as stealth delivery vehicles that are unnoticed by the immune system and liver. Furthermore, uptake of the delivery product into activated CD38-loaded (M1 population) circulating macrophages and other immunoactive leukocytes can cause rapid repolarization (back to M2 population), which rapidly reduces the levels of CD38 and the production of inflammatory cytokines. This strategy should also reduce senescent cell burden. By combining potent anti-inflammatory drugs (curcuminoids) that repolarize M1 macrophages (thus reducing CD38 levels) and promote sirtuin activity with senolytic drugs (e.g., quercetin, apigening, fisetin, luteolin), the prospect of disrupting the inflammatory cycle in chronic diseases and promoting tissue repair by enabling stem cell differentiation and proliferation is significantly maximized.

[0011] One aspect of this patent document is a transdermal formulation for enhancing systemic nitric oxide (NO) via the CD38 / NAD+ pathway, comprising: (a) an effective amount of one or more curcuminoids and an effective amount of at least one senolytic flavonoid, where the curcuminoids include, for example, one or more of curcumin, demethoxycurcumin, and bisdemethoxycurcumin, and the flavonoids from senolytic drugs include one or more of quercetin, fisetin, apigenin, luteolin, and rapamycin; (b) a polyol in an amount sufficient to dissolve an effective amount of curcuminoid and flavonoid; and optionally (c) a fatty acid, wherein the polyol and the fatty acid are in a ratio ranging from about 10:1 to about 50:1 by weight. Including, Disclosed are transdermal formulations in which the amounts of polyol and fatty acid are selected such that upon administration, an effective amount of the active agent(s) is delivered transdermally or transmucosally following topical application.

[0012] In some embodiments, the curcuminoid is curcumin. In some embodiments, the other active is at least one flavonoid from one of the following: quercetin, apigenin, fisetin, luteolin, or another small biocompatible senolytic drug such as rapamycin. In some embodiments, the formulation comprises a combination of a curcuminoid and a senolytic drug such as quercetin and apigenin.

[0013] The agents delivered via the formulations disclosed herein can enhance systemic nitric oxide levels in the endothelium by boosting NO production or releasing NO into the circulation.By upregulating systemic NO levels or delivering an effective amount of NO to the circulation, the formulations and kits disclosed herein can be applied to the treatment of various conditions or diseases. Examples include percutaneous treatment of acute and chronic inflammatory conditions, percutaneous prevention and reversal of endothelial dysfunction, reducing the risk of cytokine storm or cytokine storm phenomenon (release of abnormal or higher than normal levels of inflammatory cytokines) in patients with conditions causing underlying endothelial dysfunction (e.g., COVID-19 or SARS-CoV-2 infection), topical treatment of dermatological conditions, anti-aging skin treatments, ophthalmological conditions, aerosol-based treatment of pulmonary conditions, topical treatment of infectious diseases, treatment of red blood cells to improve storage characteristics and reverse storage damage, thus improving the safety and efficacy of stored red blood cells, loading therapeutic agents into red blood cells prior to transfusion, stabilizing red blood cells via intravenous (IV) delivery of RBC stabilizing agents, IV intervention to treat cytokine storm phenomenon and associated 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 subsequent inflammation, and loading medical sponges with therapeutic agents, for example for use in the ear, nose, mouth, rectum, and vagina. Sustained transdermal systemic delivery can also be achieved to harness the therapeutic potential of NO for a variety of chronic diseases and conditions.

[0014] The formulation further comprises polyphenols, flavonoids, stilbenoids, secosteroids, and other phytochemicals and natural products that promote the formation of NO. In some embodiments, the NO booster comprises at least one of the agents selected from the group consisting of polyphenols, flavonoids, stilbenoids, secosteroids, and natural products. In some embodiments, the curcuminoid comprises at least one of curcumin, demethoxycurcumin, bisdemethoxycurcumin, quercetin, berberine, resveratrol, and vitamin D.

[0015] In some embodiments, the polyol is a polyethylene glycol having a molecular weight ranging from 100 to about 1000. In some embodiments, the fatty acid is myristic acid.

[0016] In some embodiments, the formulation provides sustained release of the NO releasing agent from the NO booster or NO precursor (e.g., an S-nitrosothiol-containing molecule) over a period of approximately about 8, about 10, about 15, about 20, about 24, or about 48 hours.

[0017] In some embodiments, the formulation further comprises a thickening agent that maintains the transdermal formulation in a semi-solid or solid form. In some embodiments, the thickening agent is petrolatum, cocoa butter, or a polyalkylene glycol of MW greater than 2 kDa.

[0018] Another aspect of the present invention provides a kit or transdermal delivery system incorporating the transdermal formulations disclosed herein. In some embodiments, the transdermal delivery system is a kit or a nebulizer. More specifically, the present invention provides: [1] below: (a) an effective amount of a NO booster comprising one or both of an effective amount of a curcuminoid and an effective amount of at least one flavonoid, wherein the curcuminoid is selected from the group consisting of curcumin, demethoxycurcumin, and bisdemethoxycurcumin, and the at least one flavonoid is selected from quercetin, apigenin, fisetin, luteolin, and rapamycin; (b) a polyol in an amount sufficient to dissolve the effective amount of curcuminoid and the at least one flavonoid; and optionally (c) a fatty acid, wherein the polyol and the fatty acid are in a ratio ranging from about 10:1 to about 50:1 by weight. A transdermal formulation comprising: the amounts of the polyol and the fatty acid are selected such that upon administration an effective amount of the NO booster is delivered transdermally; Transdermal formulations; [2] The transdermal formulation of [1], wherein the curcuminoid is curcumin; [3] A transdermal formulation of [1], wherein the formulation comprises curcumin, and at least one of quercetin and apigenin; [4] The transdermal formulation of [1], wherein the curcuminoid and the at least one flavonoid are in a ratio ranging from about 1:1 to about 10:1; [5] The transdermal formulation of [1], wherein the formulation contains quercetin and apigenin in a ratio ranging from about 1:1 to about 10:1; [6] The transdermal formulation of [1], wherein the fatty acid is myristic acid; [7] The transdermal formulation of [1], wherein the polyol is selected from the group consisting of polyethylene glycol, polypropylene glycol, ethylene glycol, propylene glycol, and glycerol; [8] The transdermal formulation of [1], wherein the polyol is a polyethylene glycol having a molecular weight in the range of 200 to about 600; [9] A transdermal formulation of [1] comprising curcuminoids in the range of about 3% by weight to about 10% by weight;

[10] a transdermal formulation of [1], wherein the amounts of the curcuminoid and the polyol are selected to provide a sustained release of the curcuminoid over a period of about 15 hours;

[11] The transdermal formulation of

[10] , wherein the polyol and the curcuminoid are in a ratio ranging from about 8:1 to about 12:1;

[12] 1. Use of a transdermal formulation for the manufacture of a medicament for treating a disease or condition associated with elevated CD38 levels, comprising: The transdermal formulation comprises: (a) an effective amount of a NO booster comprising one or both of an effective amount of a curcuminoid and an effective amount of at least one flavonoid, wherein the curcuminoid is selected from the group consisting of curcumin, demethoxycurcumin, and bisdemethoxycurcumin; (b) a polyol in an amount sufficient to dissolve the effective amount of curcuminoid and the at least one flavonoid; and optionally (c) a fatty acid, wherein the polyol and the fatty acid are in a ratio ranging from about 10:1 to about 50:1 by weight. Including, the amounts of the polyol and the fatty acid are selected such that upon administration an effective amount of the NO booster is delivered transdermally; use;

[13]

[12] , wherein the disease or condition is selected from lupus, rheumatoid arthritis, multiple sclerosis, leukemia, multiple myeloma, neurodegenerative diseases, age-related physical and cognitive decline, chronic and acute inflammatory conditions, renal failure, coronary artery disease, heart failure, stroke, peripheral artery disease, myocardial infarction, diabetes, hypertension, neuropathic pain, osteoarthritis, anemia of chronic disease, ALS, Parkinkinson's, ischemia-reperfusion injury, hypoxia-reoxygenation injury, transfusion-induced injury, radiation-induced injury including dermatitis, and neuroinflammation;

[14]

[12] , wherein the disease or condition is selected from the group consisting of dementia, Alzheimer's disease (AD), and Parkinson's disease;

[15]

[12] , wherein the disease or condition is selected from the group consisting of lupus, rheumatoid arthritis, multiple sclerosis, leukemia, and multiple myeloma;

[16]

[12] wherein the disease or condition is type 2 diabetes;

[17] 1. Use of a transdermal formulation for the manufacture of a medicament for reducing CD38 activity, comprising: The transdermal formulation comprises: (a) an effective amount of a NO booster comprising one or both of an effective amount of a curcuminoid and an effective amount of at least one flavonoid, wherein the curcuminoid is selected from the group consisting of curcumin, demethoxycurcumin, and bisdemethoxycurcumin; (b) a polyol in an amount sufficient to dissolve the effective amount of curcuminoid and the at least one flavonoid; and optionally (c) a fatty acid, wherein the polyol and the fatty acid are in a ratio ranging from about 10:1 to about 50:1 by weight. Including, the amounts of the polyol and the fatty acid are selected such that upon administration an effective amount of the NO booster is delivered transdermally; use;

[18]

[17] , wherein the CD38 is present on a cell;

[19]

[18] , wherein the contacting occurs in vitro;

[20] The use of

[18] , wherein the contacting occurs in vivo; [twenty one]

[12] , wherein the formulation comprises curcumin and at least one of quercetin and apigenin; [twenty two] The use of

[21] , wherein the polyol is a polyethylene glycol having a molecular weight in the range of 200 to about 600; [twenty three]

[17] , wherein the formulation comprises curcumin and at least one of quercetin and apigenin; [twenty four] The use of

[23] , wherein the polyol is a polyethylene glycol having a molecular weight in the range of 200 to about 600. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0019] Detailed Description of the Invention The embodiments of the present disclosure provide transdermal formulations and methods that enhance systemic NO levels.Since NO has known systemic benefits, including the ability to reverse inflammation, prevent and reverse endothelial dysfunction, repolarize activated macrophages, inactivate activated platelets, protect the endothelial lining of blood vessels, and have antibacterial / antiviral activity, the formulations of the present disclosure can be applied to the treatment of various diseases and conditions.The treatment is based on the transdermal delivery of agents that can improve nitric oxide production in endothelium or directly release NO from suitable S-nitrosothiol-containing molecules.The formulations disclosed herein can promote the production of NO in endothelium and reduce ROS levels through the reduction of CD38 levels.

[0020] The transdermal formulations described herein have the advantage of reduced side effects compared to conventional oral administration of active agents.For example, oral curcumin can cause GI upset (hypermotility, increased acid production in the stomach), especially when used chronically.Delivery of drugs on the skin or mucosa bypasses the GI tract and achieves the desired therapeutic effect while minimizing side effects.

[0021] The following text may refer to or exemplify specific embodiments of the formulation, kit or method for treating or preventing a disease, but is not intended to limit the scope of the formulation, kit or method to such specific references or examples. Various modifications may be made by those skilled in the art, taking into account practical and economic considerations, such as the specific form of the formulation and the amount or frequency of administration of the formulation for treating or preventing a disease or condition.

[0022] As used herein, the articles "a" and "an" refer to "one or more" or "at least one," unless otherwise indicated. That is, reference to any element or component of an embodiment by the indefinite article "a" or "an" does not exclude the possibility that multiple elements or components are present.

[0023] As used herein, the term "about" refers to the referenced numerical indication plus or minus 10% of the referenced numerical indication. In some embodiments, "about" refers to the referenced numerical indication plus or minus 5% of the referenced numerical indication.

[0024] The term "drug" or "active agent" refers to a molecule or compound that prevents, alleviates or improves symptoms of a disease, prolongs the life of the subject being treated, or achieves a desired / acceptable medical or hygienic condition. The agent in the NO booster increases the systemic production of NO in the subject. The agent in the NO precursor releases NO or undergoes a reaction to produce another agent, which releases NO after being delivered transdermally into the blood circulation.

[0025] The term "body cavity" includes any opening on the body of a subject and / or the surface area within an opening. Non-limiting examples of body cavities include the nose, sinuses, mouth, ears, rectum, vagina, open wounds, sores, buccal cavity, and mucosal surfaces (e.g., gums).

[0026] Term C 1~30Alkyl includes branched or unbranched alkyl groups having any number of carbons ranging from 1 to 30. Non-limiting examples include methyl, ethyl, propyl, and butyl.

[0027] The term “cytokine storm” refers to the abnormal systemic release of dysregulated proinflammatory cytokines that leads to disease, also known as “cytokine release syndrome” or “inflammatory cascade.” Often, a cytokine storm or cascade is referred to as being part of a sequence because one cytokine typically leads to the production of multiple other cytokines that can enhance and amplify the immune response. In general, these proinflammatory mediators are divided into two subgroups: early and late mediators. For example, early mediators such as tumor necrosis factor, interleukin-1, and interleukin-6 are not adequate therapeutic targets to re-establish homeostatic balance because they dissipate within the time frame that a patient visits a clinic to receive medical care. In contrast, so-called “late mediators” have been targeted because it is during this later “inflammatory cascade” that patients realize they have become ill.

[0028] The term "inflammatory disease or disorder" may refer to any disease, disorder, or syndrome in which an excessive or unregulated inflammatory response results in a transient inflammatory state, damage to host tissue, or loss of tissue function. "Inflammatory disease" also refers to pathological conditions mediated by granulocyte influx and / or neutrophil chemotaxis, as well as transient inflammatory states including leaky gut syndrome and "brain fog" resulting from chemotherapy.

[0029] The term "long covid" refers to the adverse effects or symptoms caused by Covid that become evident after one appears to have recovered from the primary infection. Non-limiting examples of long covid symptoms include brain fog, fatigue, pain, clotting problems, myocarditis, and edema.

[0030] The term "NO booster" refers to an agent or mixture of agents that increases the systemic production of NO in a subject. The NO booster itself does not release NO.

[0031] The term "NO precursor" refers to a drug or mixture that releases NO directly or indirectly through a derivative. The NO precursor can be or include a drug that contains a NO-releasing moiety, which is delivered transdermally into the blood circulation of a subject and then releases NO. Non-limiting examples of such NO-releasing agents include S-nitroso-glutathione (GSNO), S-nitroso-N-acetylcysteine ​​(SNAC), S-nitroso-N-acetylpenicillamine (SNAP), and S-nitroso-human serum albumin (SNO-HAS). Alternatively, the NO precursor can include a drug that produces a derivative that contains a NO-releasing moiety, which releases NO after being delivered transdermally into the blood circulation. Non-limiting examples of drugs that produce NO-releasing derivatives include glutathione, N-acetylcysteine ​​(NAC), N-acetylpenicillamine, and cysteine, which can be nitrosated at the thiol group to produce S-nitrosothiol-containing derivatives.

[0032] The term "semi-solid" refers to a flexible, deformable solid form. Free-flowing liquids and hard solid forms are excluded from semi-solids. Non-limiting examples include gels, ointments, creams, emulsions, microemulsions, nanoemulsions, pastes, balms, lotions, and mousses.

[0033] The term "subject" includes any animal, but preferably includes a mammal, such as a human, non-human primate, dog, cat, horse, cow, or rodent. More preferably, the subject is a human.

[0034] The term "pharmaceutical acceptable salt" refers to a salt of the compound of the present invention that is pharmaceutically acceptable and has the desired pharmacological activity. Non-limiting examples of such salts include inorganic acids such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, and phosphoric acid; or 1,2-ethanedisulfonic acid, 2-hydroxyethanesulfonic acid, 2-naphthalenesulfonic acid, 3-phenylpropionic acid, 4,4'-methylenebis(3-hydroxy-2-ene-1-carboxylic acid), 4-methylbicyclo[2.2.2]oct-2-ene-1-carboxylic acid, acetic acid, aliphatic mono- and dicarboxylic acids, aliphatic sulfuric acid, aromatic sulfuric acid, benzenesulfonic acid, benzoic acid, camphorsulfonic acid, carbonic acid, cinnamic acid, citric acid, cyclopentanepropionic acid, and the like. Pharmaceutically acceptable salts include acid addition salts formed with organic acids such as ethanesulfonic acid, fumaric acid, glucoheptonic acid, gluconic acid, glutamic acid, glycolic acid, heptanoic acid, hexanoic acid, hydroxynaphthoic acid, lactic acid, lauryl sulfuric acid, maleic acid, malic acid, malonic acid, mandelic acid, methanesulfonic acid, muconic acid, o-(4-hydroxybenzoyl)benzoic acid, oxalic acid, p-chlorobenzenesulfonic acid, phenyl-substituted alkanoic acid, propionic acid, p-toluenesulfonic acid, pyruvic acid, salicylic acid, stearic acid, succinic acid, tartaric acid, tertiary butyl acetic acid, and trimethyl acetic acid. Pharmaceutically acceptable salts also include base addition salts that can be formed when acidic protons present can react with inorganic or organic bases. Acceptable inorganic bases include sodium hydroxide, sodium carbonate, potassium hydroxide, aluminum hydroxide, and calcium hydroxide. Non-limiting examples of acceptable organic bases include ethanolamine, diethanolamine, triethanolamine, tromethamine, and N-methylglucamine. It should be recognized that the particular anion or cation forming a part of any salt of this invention is not critical, so long as the salt, as a whole, is pharmacologically acceptable.Further examples of pharma- ceutically acceptable salts and methods for their preparation and use are provided in Handbook of Pharmaceutical Salts: Properties, and Use (PH Stahl & CG Wermuth eds., Verlag Helvetica Chimica Acta, 2002).

[0035] The term "therapeutically effective amount" or "effective amount" refers to an amount of an active agent effective to prevent, alleviate or ameliorate symptoms of a disease, prolong the survival of the subject being treated, or achieve a desirable / acceptable medical or hygienic condition. Determination of a therapeutically effective amount or an effective amount is well within the capabilities of those skilled in the art, especially in light of the detailed disclosure provided herein.

[0036] The term "treat" or "treatment" of any disease or condition, in some embodiments, refers to improving the disease or disorder (i.e., preventing or reducing the onset of the disease or at least one of its clinical symptoms). In some embodiments, "treat" or "treatment" refers to improving at least one physical parameter that may not be discernible by the subject. In some embodiments, "treat" or "treatment" refers to modulating the disease or disorder physically (e.g., stabilization of discernible symptoms), physiologically (e.g., stabilization of physical parameters), or both. In some embodiments, "treat" or "treatment" refers to delaying the onset of the disease or disorder, or even preventing it. For example, treatment can be a "prophylactic treatment" and should be interpreted as any treatment modality used to prevent the progression of a disease or used for prophylactic purposes for those at risk of developing a condition.

[0037] The term "transdermal" or "transdermally" refers to the delivery, administration or application of a formulation containing an active agent by direct contact with skin or mucosa, followed by the transport of the agent through the skin or mucosa into the blood circulation of a subject.Such delivery, transport, administration or application is also known to include dermal, transdermal, transmucosal and buccal routes.As used herein, "dermal" includes skin and mucosa, including oral, buccal, nasal, rectal and vaginal mucosa.In some embodiments, this term also refers to the delivery and transport of an agent through cell walls (e.g. red blood cell walls) into cells.

[0038] The term "transdermal formulation" refers to a composition or formulation of a drug that upon application to the skin or mucosa delivers the drug or drug derivative (e.g., an S-nitrosothiol-containing molecule derived from a thiol-containing molecule) across the skin or mucosa (or any other surface as noted above). The transdermal formulation may be in the form of a solution, suspension, gel, ointment, cream, emulsion, microemulsion, nanoemulsion, paste, balm, magma, lotion, mousse, wax, or liposome. A kit or transdermal delivery system incorporating a transdermal formulation may be in the form of, for example, a patch, swab, nebulizer, sprayer, sponge, or pouch.

[0039] Transdermal formulations An aspect of the present disclosure provides a transdermal formulation that delivers a therapeutically effective amount of a drug transdermally. The formulation generally comprises the following: (a) an effective amount of an active agent, such as a CD38 inhibitor, a Sirtuin-1 (SIRT1) activator, a NO booster and / or a NO precursor; NO boosters increase systemic production of NO, and The NO precursor comprises or induces an NO releasing agent; an effective amount of an active agent; (b) a solvent in an amount sufficient to dissolve an effective amount of the NO booster or NO precursor; and optionally (c) Fatty acids Including, When administered, an effective amount of the active agent (e.g., sirtuin-1 (SIRT1) activator, NO booster and / or NO releaser) is delivered transdermally. In some embodiments, the active agent is delivered into the blood vessel so that the active agent enters the systemic circulation from the artery or vein. In some embodiments, the active agent is delivered into the deeper layers of the skin (e.g., the upper or lower epidermal layers, below the stratum corneum).

[0040] In some embodiments, the agent in the transdermal formulation is flavonoid or curcuminoid, or a combination thereof.One or more flavonoids and / or one or more curcuminoids can be included in the formulation.For example, the formulation can contain one or two flavonoids, such as quercetin and apigenin, optionally combined with curcumin, and each of the components or constituents is in a therapeutically effective amount.

[0041] In some embodiments, the transdermal formulation comprises one or more flavonoids or curcuminoids. Non-limiting examples include a combination of quercetin and apigenin, a combination of quercetin and curcumin, a combination of curcumin and apigenin, and a combination of quercetin, apigenin and curcumin. Whether the combination contains one or more flavonoids or one or more curcuminoids, the weight ratio of each of the flavonoids to each of the curcuminoids can independently range from about 1:100 to about 100:1, about 5:100 to about 100:5, 1:10 to about 10:1, 2:10 to about 10:2, 3:10 to about 10:3, 4:10 to about 10:4, 5:10 to about 10:5, 6:10 to about 10:6, or 8:10 to about 10:8. Non-limiting examples of ratios include about 100:1, about 80:1, about 50:1, about 40:1, about 30:1, about 20:1, about 10:1, about 8:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:8, about 1:10, about 1:20, about 1:30, about 1:40, about 1:50, about 1:60, about 1:80, about 1:100, and any range between any two of the foregoing values. In some embodiments, the combination contains curcumin and one or both of quercetin and apigenin, wherein the mass ratio of curcumin to each of quercetin and apigenin, independently, ranges from about 1:5 to about 10:1, from about 1:5 to about 3:1, from about 1:2 to about 3:1, or from about 1:2 to about 2:1.

[0042] When the formulation contains two or more flavonoids (e.g., quercetin, apigenin, fisetin, luteolin, and rapamycin), the weight ratio of the two flavonoids can, independently, be in the range of about 1:100 to about 100:1, about 5:100 to about 100:5, 1:10 to about 10:1, 2:10 to about 10:2, 3:10 to about 10:3, 4:10 to about 10:4, 5:10 to about 10:5, 6:10 to about 10:6, or 8:10 to about 10:8. Non-limiting examples of ratios include about 100:1, about 80:1, about 50:1, about 40:1, about 30:1, about 20:1, about 10:1, about 8:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:8, about 1:10, about 1:20, about 1:30, about 1:40, about 1:50, about 1:60, about 1:80, about 1:100, and any range between any two of the foregoing values. In some embodiments, the formulation comprises quercetin and apigenin in a ratio ranging from about 1:1 to about 10:1, from about 2:1 to about 10:1, from about 5:1 to about 10:1, from about 1:1 to about 1:10, from about 1:1 to about 1:5, or from about 1:1 to about 1:2.

[0043] The level of NO in a subject can be measured using known techniques, for example, as disclosed in US Patent Nos. 9,044,182 and 8,425,428.In some embodiments, the effective amount is sufficient to have a measurable effect on diseases, including, for example, hypertension, inflammation, osteoarthritis, endothelial dysfunction, dermatological conditions, ophthalmological conditions, bacterial infections, viral infections, ischemia-reperfusion injury, hypoxia-reoxygenation injury, cytokine storm phenomenon, cerebral malaria, Chagas disease, and hemoglobinopathies, such as sickle cell disease and HbE / β thalassemia, type 2 diabetes, and lupus.In some embodiments, the effective amount is sufficient to have a measurable positive effect on blood flow and / or vasodilation, and / or a measurable negative effect on blood pressure.In some embodiments, the effect on blood flow and / or vasodilation is observed locally to the site of topical application. In some embodiments, the effective amount is an amount sufficient to have a measurable effect on inflammatory diseases, such as inflammatory dermatoses, inflammatory bowel disease, and inflammation of the systemic vasculature, including the blood-brain barrier, caused by chemotherapy, after treatment with the formulation, as evidenced by appropriate clinical parameters, such as improvement in Physician's Global Assessment.In some embodiments, the effective amount is an amount sufficient to obtain a systemic or local level of nitric oxide to have a desired effect, such as to have a measurable positive effect on blood flow and / or vasodilation, a measurable negative effect on blood pressure, and / or to have a measurable effect on inflammatory dermatoses, such as inflammatory dermatoses, as evidenced by appropriate clinical parameters.

[0044] Non-limiting examples of SIRT1 activators, active agents include polyphenols, flavonoids, stilbenoids, secosteroids, and other phytochemicals or natural products that promote the formation of NO.

[0045] After the NO booster or NO precursor is delivered transdermally, it will result in the production of NO in the subject's body. The NO booster or NO precursor is in an amount effective to increase NO systemically or locally to a level high enough to achieve the purpose of treating a disease or condition. In some embodiments, the NO booster comprises a polyphenol, a flavonoid, a stilbenoid, a secosteroid, or a natural product that promotes NO production. In some embodiments, the NO precursor comprises an S-nitrosothiol-containing molecule, or a thiol-containing molecule and a nitrite source. In some embodiments, the NO booster comprises one or more of curcuminoids, flavonoids, berberine, resveratrol, a vitamin D source, and their pharma-ceutically acceptable salts and derivatives. Curcuminoids are linear diarylheptanoids, and include, for example, curcumin, demethoxycurcumin, and bisdemethoxycurcumin. Flavonoids have a 3-hydroxyflavone backbone, and include, for example, 3-hydroxyflavone, azaleatin, fisetin, galangin, gossypetin, kaempferide, kaempferol, isorhamnetin, morin, myricetin, natudaidin, pachypodol, quercetin, rhamnagin, and rhamnetin. Non-limiting examples of vitamin D sources include vitamin D2 and vitamin D3, and any precursor of vitamin D. Non-limiting examples of polyphenols include plant extracts, brazilin, and theaflavins (e.g., theaflavin (TF-1), theaflavin-3-gallate (TF-2a), theaflavin-3'-gallate (TF-2b), and theaflavin-3,3'-digallate (TF-3)).

[0046] In some embodiments, the NO booster consists essentially of curcumin, demethoxycurcumin, bisdemethoxycurcumin, quercetin, berberine, resveratrol, a vitamin D source, and any combination thereof.

[0047] In some embodiments, the formulation contains both NO booster and NO precursor.For example, the combination of curcumin and NO releaser (e.g., S-nitrosothiol-containing molecule or thiol-containing drug) in polyol / fatty acid system can be a powerful formulation for treating localized inflammation and infection, while providing the systemic benefits of curcumin to control systemic inflammation.

[0048] Additional examples of curcuminoids include methylcurcumin, demethoxycurcumin, bisdemethoxycurcumin, sodium curcuminate, dibenzoylmethane, acetylcurcumin, feruloylmethane, tetrahydrocurcumin, 1,7-bis(4-hydroxy-3-methoxyphenyl)-1,6-heptadiene-3,5-dione (curcumin 1), 1,7-bis(piperonyl)-1,6-heptadiene-3,5-dione (piperonylcurcumin), 1,7-bis(2-hydroxynaphthyl)-1,6-heptadiene-2,5-dione (2-hydroxyl naphthylcurcumin), and 1,1-bis(phenyl)-1,3,8,10 undecatetraene-5,7-dione. In some embodiments, the NO booster is curcumin or a synthetic curcumin that is 80%, 85%, 90% or 98% pure diferuloylmethane.

[0049] In some embodiments, the transdermal formulation comprises one or more curcuminoids and, optionally, one or more of polyphenols, flavonoids, stilbenoids, secosteroids, or natural products that promote NO production as active ingredients for treating a disease or condition. In some embodiments, the transdermal formulation comprises one or both of curcumin and quercetin and, optionally, one or more of polyphenols, flavonoids, stilbenoids, and secosteroids.

[0050] The NO precursor is either an S-nitrosothiol-containing molecule or a mixture containing a thiol-containing molecule and a nitrite source. When contacted with an acid source, the nitrite source produces nitrous acid, which can then nitrosate the reactive thiol of the thiol-containing molecule. The S-nitrosothiol-containing molecule releases NO to a subject in need thereof.

[0051] A variety of thiol-containing molecules can be used as precursors. Examples include glutathione, N-acetylcysteine ​​(NAC), N-acetylpenicillamine, cysteine, and their derivatives. The amino group of either cysteine ​​or NAC can be converted to an acetyl or other carbonyl of different carbon length (e.g., COC 2~30 By adjusting the length of the carbon chain, the solubility and lipophilicity of the molecule can be modified. Similarly, the carboxy group of cysteine ​​can be acetylated with an ester (e.g., an ethyl ester, or other substituted or unsubstituted C 3~30 alkyl esters) or amides having an NR2 moiety (wherein each R is independently H or other substituted or unsubstituted C 3~30 The carbon chain can be converted to an alkyl group, which is an alkyl group. Varying the carbon chain allows for tuning of the molecule's properties.

[0052] Various inorganic compounds can serve as the nitrite source. Non-limiting examples of nitrite sources include alkali metal nitrites, alkaline earth metal nitrites, transition metal nitrites, and ammonium nitrite. In some embodiments, the nitrite source is potassium nitrite, sodium nitrite, rubidium nitrite, strontium nitrite, barium nitrite, calcium nitrite, copper nitrite, zinc nitrite, or mixtures thereof. Nitrite can also include natural sources such as lettuce and spinach extracts. In some embodiments, the nitrite source is saturated in a polyol solvent. The nitrite source can also be comprised of nanoparticles loaded with nitrite.

[0053] Nitrite-loaded nanoparticles can be prepared by techniques known in the art, including, for example, the procedure reported in U.S. Patent No. 8,333,997, the entire disclosure of which is incorporated herein by reference. To limit NO release from nanoparticles during manufacture, the medium should maintain a pH above about 7.5 throughout preparation. The nitrite-loaded nanoparticles can then be mixed with a solvent system of polyol and fatty acid (e.g., PEG400 and myristic acid) and remain stable until exposed to an aqueous environment. In the presence of thiol-containing molecules, the nitrite-loaded nanoparticles will allow the formation of S-nitrosothiols when the mixture is exposed to an acid source or a slightly acidic aqueous environment on the skin.

[0054] The use of nitrite-loaded nanoparticles allows for the use of high concentrations of nitrite under hydrophobic conditions. The combination of nitrite-loaded nanoparticles with a solvent system of polyols and fatty acids (e.g., PEG400 and myristic acid) allows for a stable mixture that releases NO and S-nitrosates thiols when exposed to an aqueous environment, regardless of other deliverables contained therein. There is no release of nitrite or production of NO in the viscous solvent until water or an acid source is introduced.

[0055] The acid source can be packaged separately from the mixture containing the thiol-containing molecule and the nitrite source, and mixed with the nitrite salt before administration.For example, the nitrite source and the acid source can be packaged separately in a permeable or frangible pouch.The amount and concentration of the acid can be adjusted according to the amount of other drugs and the nature of the acid.Non-limiting examples of the acid include acetic acid, oxalic acid, and citric acid.

[0056] Non-limiting examples of S-nitrosothiol-containing molecules include S-nitroso-glutathione (GSNO), S-nitroso-N-acetylcysteine ​​(SNAC), S-nitroso-N-acetylpenicillamine (SNAP), and S-nitroso-human serum albumin (SNO-HAS). As with thiol-containing molecules, these S-nitrosothiol-containing molecules can be modified by varying the carbon chains in the respective ester, amide, or N-acyl moieties to fine-tune their properties.

[0057] A solvent such as a polyol suitable for the delivery system allows for high concentrations of poorly soluble drugs. In addition, it should be biocompatible and profiled as safe for biomedical applications. Furthermore, it ideally facilitates skin and mucosal penetration to allow transdermal delivery. Non-limiting examples of polyols include polyethylene glycol, polypropylene glycol, ethylene glycol, propylene glycol, and glycerol. In some embodiments, the polyol is polyethylene glycol (PEG). In some embodiments, the PEG has a molecular weight in the range of about 100 to about 2000, about 100 to about 1000, about 100 to about 800, about 100 to about 600, about 200 to about 600, or about 200 to about 400 daltons. In some embodiments, the formulation is substantially free of water.

[0058] In some embodiments, the solvent of the formulation consists essentially of polyol. In some embodiments, the formulation may contain one or more additional solvents. Non-limiting examples include mineral oil, petrolatum, castor oil, eugenol, menthol, cineole, or essential oils such as rose oil, n-methylpyrrolidone, vegetable oils, oleyl alcohol, dipropylene glycol, polyoxyethylene derivatives of sorbitan esters, saturated polyglycolized C 8~10These include glycerides, polyoxyethylated fatty acid glycerides, oleic acid, dimethyl sulfoxide (DMSO), fatty alcohols, isopropyl myristate (IPM), triacetin, ethyl oleate, isostearic acid, medium chain fatty acids and other fats, and mixtures thereof. In addition to dissolving the drug, these solvents can also function as plasticizers, so that the formulation can be flexible, stretchable, moldable and / or otherwise skin-friendly.

[0059] In some embodiments, the polyol solvent is a low molecular weight polyethylene glycol (PEG) having a molecular weight in the range of about 50 to about 2000, about 50 to about 1000, about 100 to about 1000, about 100 to about 800, about 100 to about 700, about 100 to about 600, about 200 to about 800, about 200 to about 600, or about 200 to about 400 Daltons. Non-limiting examples of molecular weights of polyol solvents include about 100, about 200, about 300, about 400, about 500, about 600, about 800, and about 1000. Short chain PEG molecules such as PEG200 and PEG400, which are liquid at ambient temperature, are particularly useful.

[0060] Fatty acids serve as permeation enhancers.Non-limiting examples of fatty acids include myristoleic acid, palmitoleic acid, sapienic acid, oleic acid, elaidic acid, vaccenic acid, linoleic acid, linoelaidic acid, alpha-linolenic acid, arachidonic acid, eicosapentaenoic acid, erucic acid, docosahexaenoic acid, caprylic acid, capric acid, lauric acid, myristic acid, palmitic acid, stearic acid, arachidic acid, behenic acid, lignoceric acid, cerotic acid, myristic acid, and any combination thereof.In some embodiments, the fatty acid is myristic acid.Medium-sized fatty acids such as myristic acid and / or other fatty acids of similar size / molecular weight are particularly useful.In some embodiments, the formulation does not contain fatty acid or contains only trace or minor amounts of fatty acid.

[0061] In some embodiments, the permeation enhancer is essentially composed of fatty acid or its ester.In some embodiments, the formulation may contain one or more additional permeation enhancers.Non-limiting examples include surfactants, alcohols, fatty alcohols and glycols, esters, fatty acid esters and fatty alcohol esters, esters of long-chain fatty acids with methyl, ethyl, isopropyl alcohol, esters of fatty alcohols with acetic acid, lactic acid and oleic acid, diethanolamines, essential oils, terpenes and terpenoids, amides, urea, polyoxyethylene fatty alcohol ethers, polyoxyethylene fatty acid esters, sulfoxides, ether alcohols, pyrrolidones, transcarbam, capsaicin derivatives, dimethyl amino acid esters, peptides, iminosulfurans, dicarboxylic acid esters, nanocarriers, triglycerides, carbohydrates, phospholipids, alone or in combination.

[0062] By adjusting the amount and ratio of polyol, fatty acid, and NO booster or NO precursor, the solubility of fatty acid and drug in NO booster or NO precursor, as well as the physical state of the formulation (e.g., liquid or gel or semi-solid) and the release profile of active drug can be controlled. The ratio of polyol to fatty acid affects the form of the solution, and generally ranges from about 5:1 to about 500:1, about 5:1 to about 100:1, about 1:1 to about 100:1, about 20:1 to about 100:1, about 30:1 to about 100:1, about 20:1 to about 80:1, about 20:1 to about 60:1, about 50:1 to about 10:1, or about 30:1 to about 50:1 about 40:1 to about 10:1, about 40:1 to about 10:1, about 20:1 to about 60:1, about 20:1 to about 15:1, about 18:1 to about 12:1 by weight. In some embodiments, the concentration of the fatty acid in the polyol ranges from about 0.01 M to about 1 M, about 0.01 M to about 0.8 M, about 0.01 M to about 0.6 M, about 0.01 M to about 0.4 M, about 0.01 M to about 0.2 M, about 0.01 M to about 0.15 M, about 0.01 M to about 0.1 M, about 0.02 M to about 0.2 M, about 0.02 M to about 0.1 M, about 0.04 M to about 0.08 M, or about 0.06 M to about 0.1 M. In further exemplary embodiments, the concentration of the fatty acid in the polyol is about 0.01 M, about 0.02 M, about 0.03 M, about 0.04 M, about 0.06 M, about 0.08 M, about 0.1 M, or about 0.12 M. In some embodiments, the fatty acid is saturated in the polyol. In some embodiments, the polyol is PEG. In some embodiments, the fatty acid is myristic acid.

[0063] The range and amount of polyol and fatty acid are as described above and can be modified by those skilled in the art in view of practical needs without undue experimentation. In some embodiments, the fatty acid is selected from myristoleic acid, palmitoleic acid, sapienic acid, oleic acid, elaidic acid, vaccenic acid, linoleic acid, linoelaidic acid, α-linolenic acid, arachidonic acid, eicosapentaenoic acid, erucic acid, docosahexaenoic acid, caprylic acid, capric acid, lauric acid, myristic acid, palmitic acid, stearic acid, arachidic acid, behenic acid, lignoceric acid, cerotic acid, and any combination thereof. In some embodiments, the polyol is selected from the group consisting of polyethylene glycol, polypropylene glycol, ethylene glycol, propylene glycol, glycerol, and combinations thereof. In some embodiments, the polyol is a polyethylene glycol having a molecular weight ranging from 200 to about 600. In some embodiments, the polyol and fatty acid are in a ratio ranging from about 5:1 to about 100:1 by weight.

[0064] The fatty acid may range from about 0.1% to about 30%, about 0.5% to about 20%, about 1% to about 15%, about 1% to about 10%, about 1% to about 5%, about 2% to about 8%, or about 4% to about 8% of the total weight of the NO booster or NO precursor, polyol, and fatty acid (if present) or the total weight of the formulation. Non-limiting examples of amounts of fatty acid include about 1%, about 3%, about 5%, about 7%, about 8%, or about 10% by weight.

[0065] Water can cause aggregation and particulate formation. In some embodiments, the transdermal formulation is anhydrous or substantially free of water. Minimizing or removing water from the formulation helps maintain uniform distribution of fatty acid and / or active ingredient (e.g., NO booster or NO precursor) in polyol (e.g., PEG). In some embodiments, the water in the transdermal formulation is less than 5% by weight, less than 2% by weight, less than 1% by weight, less than 0.5% by weight, less than 0.1% by weight, or less than 0.01% by weight.

[0066] In some embodiments, the transdermal formulation includes at least one water repellent, also called a water repellant. Examples of water repellents include silicones, such as cyclomethicone, dimethicone, simethicone, C 26~28 Alkyl dimethicone, C 26~28 These include alkylmethicone, polyphenylsisquioxane, trimethylsiloxysilicate, and the crosspolymer of cyclopentasiloxane and dimethicone / vinyltrimethylsiloxysilicate, and blends thereof.Water repellent can be particularly useful in the embodiment where topical vehicle is used with water-reactive agent, for example, nitric oxide releasing agent (e.g., diazenium dialate or sodium nitrite) that releases nitric oxide in the presence of water.In other cases, such as when active agent is not water sensitive, water repellent may or may not be included.

[0067] Depending on the therapeutic goal of the formulation, the active agent (e.g., CD38 inhibitor, NO booster or NO precursor or mixtures thereof) in the formulation may range from about 0.05% to about 80%, about 0.05% to about 50%, about 0.05% to about 35%, about 0.05% to about 30%, about 0.05% to about 20%, about 0.05% to about 10%, about 0.1% to about 20%, about 0.1% to about 10%, about 0.1% to about 5%, about 0.5% to about 20%, about 0.5% to about 10%, about 0.5% to about 5%, about 1% to about 20%, about 1% to about 10%, or about 1% to about 5% of the total weight of the NO booster or NO precursor, polyol, and fatty acid (if present) or the total weight of the formulation. Non-limiting examples of the amount of active agent in the formulation include about 1%, about 3%, about 5%, about 7%, about 8%, about 10%, about 12%, and about 15% by weight of the formulation. In some embodiments, the amount of active agent in the formulation or in a dosage unit of the formulation is independently in the range of about 0.001 mg to about 20 g, about 0.002 mg to about 20 g, about 0.004 mg to about 20 g, about 0.006 mg to about 20 g, about 0.008 mg to about 20 g, about 0.01 mg to about 20 g, about 0.05 mg to about 20 g, about 0.1 mg to about 20 g, about 0.1 mg to about 5 g, about 0.1 mg to about 2 g, about 0.1 mg to about 1 g, about 1 mg to about 5 g, about 1 mg to about 1 g, about 10 mg to about 100 mg, about 5 mg to about 50 mg, or about 10 mg to about 30 mg in a dosage unit of the formulation. The dosage unit can be in physically distinct packaged forms (e.g., capsules, patches, vials). The dosage unit can also be a predetermined portion of the formulation for each individual administration. For example, a suitable amount of the formulation as a dosage unit can be removed from a container for direct topical application or for loading onto a patch or any suitable carrier prior to topical application. The amount or size of the dosage unit can be easily adjusted depending on the purpose of use and area of ​​application.Non-limiting examples of the amount of active agent in a dosage unit include about 0.001 mg, about 0.002 mg, about 0.004 mg, about 0.006 mg, about 0.008 mg, about 0.01 mg, about 0.02 mg, about 0.04 mg, about 0.06 mg, about 0.08 mg, about 0.1 mg, about 0.2 mg, about 0.4 mg, about 0.06 mg, about 0.08 mg, about 1 mg, about 2 mg, about 5 mg, about 10 mg, about 20 mg, about 30 mg, about 40 mg, about 50 mg, about 60 mg, about 80 mg, about 100 mg, about 200 mg, about 300 mg, about 400 mg, about 500 mg, about 600 mg, about 800 mg, about 1 g, about 2 g, about 5 g, about 10 g, about 15 g, about 20 g, about 25 g, about 30 g, about 35 g, about 40 g, about 50 g, about 60 g, about 80 g, about 100 g, and any range between any two of the aforementioned values. In some embodiments, each active agent in the dosage unit is independently in the range of about 0.01 to about 1 mg, about 0.01 to about 0.5 mg, or about 0.1 to about 0.5 mg. In some embodiments, the active agent is a curcuminoid selected from one, two, or three of curcumin, demethoxycurcumin, and bisdemethoxycurcumin, optionally in combination with one or more flavonoids listed above. The dosage unit can be administered once, twice, three times a day, or as needed. In some embodiments, the dosage unit is administered daily, every second day, every third day, every fourth day, every fifth day, every sixth day, every seventh day, or every tenth day.

[0068] In some embodiments, the ratio of each active agent to polyol is independently in the range of about 1:5 to about 1:100, about 1:5 to about 1:50, about 1:5 to about 1:30, about 1:5 to about 1:20, about 1:8 to about 1:15, or about 1:10 to about 1:15 by weight. In some embodiments, the polyol is PEG. In some embodiments, the fatty acid is myristic acid. In some embodiments, the formulation is substantially free of piperine. Depending on the disease or condition to be treated and the site of administration, the ratio and amount of PEG, fatty acid and active agent can be selected so that the resulting formulation is a liquid, gel or other suitable form. Additional agents may be added to control the physical state of the formulation.

[0069] In some embodiments, the transdermal formulation contains PEG, myristic acid and / or other fatty acids of comparable size / molecular weight, and one or more active agents. In some embodiments, the transdermal formulation contains PEG, myristic acid and / or other fatty acids of comparable size / molecular weight, an active agent, and a second agent. In some embodiments, the transdermal formulation contains PEG, myristic acid, and an active agent selected from at least one of curcumin, demethoxycurcumin, bisdemethoxycurcumin, quercetin, berberine, resveratrol, and vitamin D. In some embodiments, the transdermal formulation contains PEG, myristic acid, and one or more active agents selected from at least one of curcumin, demethoxycurcumin, bisdemethoxycurcumin, quercetin, berberine, resveratrol, and vitamin D, and an NO precursor or a second agent. In some embodiments, the transdermal formulation contains PEG having a molecular weight ranging from about 200 to about 500 (e.g., PEG200, PEG300, PEG400, or PEG500). The ratio of PEG to myristic acid (and / or other fatty acids of comparable size / molecular weight) ranges from about 5:1 to about 100:1 by weight (e.g., 6:1, 8:1, 10:1, 12:1, 15:1, 18:1, 20:1, 25:1, 30:1, 40:1, 50:1, 60:1, or 80:1). The ratio of PEG to NO booster ranges from about 5:1 to about 100:1 by weight (e.g., 6:1, 8:1, 10:1, 12:1, 15:1, 18:1, 20:1, 25:1, 30:1, 40:1, 50:1, 60:1, or 80:1). In some embodiments, the formulation contains curcumin. In some embodiments, the formulation contains curcumin, demethoxycurcumin, bisdemethoxycurcumin, or any combination thereof. In some embodiments, the formulation contains vitamin D. In some embodiments, the concentration of each active ingredient in the formulation ranges from about 0.01 M to about 1 M, from about 0.05 M to about 0.5 M, from about 0.05 M to about 0.3 M, or from about 0.1 M to about 0.2 M.Non-limiting examples of concentrations of an active ingredient (e.g., curcumin) in a polyol (e.g., PEG) include about 0.06 M, about 0.08 M, about 0.1 M, about 0.12 M, about 0.14 M, about 0.16 M, about 0.18 M, about 0.20 M, about 0.25 M, about 0.30 M, about 0.40 M, about 0.60 M, and about 0.80 M.

[0070] The transdermal formulation has an extended shelf life with minimal degradation of the active ingredient(s). In some embodiments, the active ingredient(s) of the formulation remain greater than 95% or greater than 99% stable for a period of at least 1 month, at least 3 months, at least 6 months, or at least 12 months. In some embodiments, the transdermal formulation comprises one or more curcuminoids, myristic acid, and PEG. The one or more curcuminoids range from about 2% to about 10%, about 3% to about 8%, or about 4% to about 6% by weight in the formulation. The myristic acid ranges from about 1% to about 10%, about 1% to about 8%, about 2% to about 8%, or about 4% to about 6% by weight in the formulation. The PEG ranges from about 60% to about 95%, about 70% to about 90%, about 80% to about 90%, or about 95% to about 90% by weight in the formulation. In some embodiments, the PEG is PEG400.

[0071] In some embodiments, the formulation further comprises a gelling or thickening agent that maintains it in a semi-solid or solid form.Non-limiting examples of gelling or thickening agents include carbomer, methylcellulose, hydroxypropylmethylcellulose, poloxamer, polyacrylic acid, alginate, chitosan, xanthan gum, gellan gum, xyloglucan, paraffin, silicone, petrolatum, cocoa butter, and high molecular weight polyalkylene glycols.Additional examples include polyethylene oxide, ammonia methacrylate, carrageenan, aqueous cellulose acetate phthalate such as CAPNF from Eastman, sodium carboxymethylcellulose, carboxypolymethylene, cellulose, cellulose acetate (microcrystalline), cellulose polymers, divinylbenzenestyrene, ethylcellulose, ethylene vinyl acetate, silicones, polyisobutylene, shellac (FMC BioPolymer), cellulose derivatives such as guar gum, guar rosin, hydroxyethylcellulose, hydroxymethylcellulose, hydroxypropylcellulose, hydroxypropylmethylcellulose, carboxymethylcellulose, and methylcellulose, hypromellose phthalate (hydroxypropylmethylcellulose phthalate), methyl acrylate, microcrystalline wax, polyvinyl alcohol, polyvinyl acetate, polyvinyl acetate phthalate such as Suretic from Colorcon, PVP ethylcellulose, polyvinylpyrrolidone (PVP), acrylates, PEG / PVP, trimethylsiloxysilicate, maleic acid / anhydride copolymer, polacrilin, poloxamer, poly glactic acid Methacrylic acid and methacrylate-based polymers such as poly(methacrylic acid) copolymers and methyl methacrylate copolymers, including poly(methacrylic acid) copolymers and methyl methacrylate copolymers, including methacrylic acid-ethyl acrylate copolymers such as BASF's Kollicoat polymers, Eudragit polymers (Eudragit (E, L, NE, RL, RS, S100)) from Rohm and Haas, esters of polyvinyl methyl ether / maleic anhydride copolymers such as Gantrez ES-425, Gantrez ES-225 from ISP, and mixtures thereof. Non-limiting examples of high molecular weight polyalkylene glycols include PEG and polypropylene glycol (PPG).The polyalkylene glycol may have a molecular weight of greater than 1k Daltons, greater than 2k Daltons, greater than 3k Daltons, greater than 4k Daltons, greater than 6k Daltons, greater than 8k Daltons, greater than 10k Daltons, greater than 15k Daltons, greater than 20k Daltons, greater than 25k Daltons, or greater than 30k Daltons. Without limiting the scope, the semi-solid formulation may be in the form of an ointment, gel, cream, emulsion, paste, lotion, or liposomal dosage form.

[0072] In some embodiments, the formulation includes a combination of small and large polyalkylene glycols with MW differences ranging from 500-5000, 1000-3000, 1000-2000, or 1500-2000 Daltons. By adjusting the ratio between two or more polyalkylene glycols, the viscosity and the rate / degree of both skin penetration and uptake by the circulation can be controlled. For example, the combination may include one or both of a PEG and a PPG with MW ranging from 100-2000, 200-2000, 400-1000, or 500-800 Daltons, respectively. The combination may also include one or both of a PEG and a PPG with a higher MW ranging from 800-5000, 1000-3000, or 1000-2000 Daltons, respectively. In further exemplary embodiments, one of the polyalkylene glycols has a MW of 100, 200, 400, 600, or 800 and the other has a MW of 1000, 1500, 2000, 2500, or 3000. In some embodiments, the combination comprises a 400 Dalton PEG and a 2000 Dalton PEG. In some embodiments, the ratio of low MW polyalkylene glycol to high MW polyalkylene glycol ranges from about 10:1 to about 1:10, from about 5:1 to about 1:5, from about 2:1 to about 1:2 by weight. Further exemplary ratios of low MW polyalkylene glycol (e.g., PEG and / or PPG) to high MW polyalkylene glycol (e.g., PEG and / or PPG) include 10:1, 8:1, 6:1, 4:1, 2:1, 1:1, 1:2, 1:4, 1:6, 1:8 and 1:10.

[0073] In some embodiments, the formulation does not contain additional therapeutic agents other than the NO booster or NO precursor.In some embodiments, the active agent in the formulation consists essentially of the NO booster and / or NO precursor described herein.In some embodiments, the formulation may contain additional therapeutic agents, including, for example, antioxidants, antibiotics, antiviral agents and / or antifungal agents.

[0074] The formulations may contain other ingredients including, for example, solubilizers, skin penetration enhancers, surfactants, cosolvents, thickening or viscosifying agents, preservatives, tonicity agents, isoosmotic agents, drug absorption enhancers, mucoadhesive polymers, non-mucoadhesive polymers, chelating agents, stabilizers, antioxidants, and mixtures thereof.

[0075] In some embodiments, the thickening agent is selected from one or more of carbomer, methylcellulose, hydroxypropylmethylcellulose, poloxamer, polyacrylic acid, alginate, chitosan, xanthan gum, gellan gum, xyloglucan, paraffin, silicone, petrolatum, and cocoa butter.

[0076] Non-limiting examples of solubilizers include, but are not limited to, diethylene glycol monoethyl ether (ethoxydiglycol, commercially available as Transcutol®) and diethylene glycol monoethyl ether oleate (Soficutol®, commercially available under the trade name Poly™); polyethylene castor oil derivatives, such as polyoxy 35 castor oil, polyoxy 40 hydrogenated castor oil; polyethylene glycols, especially low molecular weight polyethylene glycols; polyethylene glycol derivatives such as caprylic / capric glyceride (commercially available as Labrasol®); alkyl methyl sulfoxides such as DMSO; pyrrolidones such as 2-pyrrolidone and N-methyl-2-pyrrolidone; and DMA. Many solubilizers can also function as absorption enhancers. A single solubilizer can be incorporated into the formulation, or a mixture of solubilizers can be incorporated into the formulation.

[0077] Non-limiting examples of skin penetration enhancers aid in facilitating the passage of therapeutic levels of an active agent through a reasonably sized area of ​​intact skin. Suitable accelerators are well known in the art and include, for example, lower alcohols such as methanol, ethanol, and 2-propanol; alkyl methyl such as dimethylsulfoxide (DMSO), decylmethylsulfoxide (C10MSO), and tetradecylmethylsulfoxide, sulfoxides; urea; 2-pyrrolidone, N-methyl-2-pyrrolidone, and N-pyrrolidones such as -(hydroxyethyl)pyrrolidone N,N-diethyl-m-toluamide; C2-C6 alkanediols; dimethylformamide (DMF), N,N-dimethylacetamide (DMA), and miscellaneous solvents such as tetrahydrofurfuryl alcohol; and 1-substituted azacycloheptan-2-ones, particularly 1-N-dodecylazacycloheptan-2-one (laurocapram, available under the trade name Azone® from Whitby Research Incorporated, Richmond, Va.).

[0078] Among the surfactants, mention may be made, for example, of polyethoxylated glycerides, polysorbates, poloxamers, sodium lauryl sulfate, phospholipids such as phosphatidylcholine or phosphatidylglycerol and derivatives thereof, polyoxyethylated hydrogenated castor oil, polyoxyethylated fatty acids, mixtures of mono-, di- and triglycerides of fatty acids, optionally polyoxyethylated, and mixtures thereof.

[0079] Among the preservatives, mention may be made, for example, of benzalkonium chloride, boric acid, benzoic acid, C1-4 alkyl esters of p-hydroxybenzoic acid, chlorobutanol, benzyl alcohol, phenylethyl alcohol, organometallic derivatives of mercury, polyquaterniums such as Polyquaternium 1, and mixtures thereof.

[0080] Among the isotonic and isotonic agents, there may be mentioned, for example, inorganic salts such as sodium chloride, glucose, trehalose, mannitol, amino acids, and mixtures thereof.

[0081] Among the mucoadhesive polymers, for example, hyaluronic acid, polygalacturonic acid, polyacrylic acid, carboxymethyl amylose, carboxymethyl chitin, chondroitin sulfate, methylcellulose, gelatin, hydroxymethylcellulose, hydroxypropylcellulose, hydroxypropylmethylcellulose, sodium carboxymethylcellulose, xanthan gum, chitosan, carbopol, polycarbophil, gellan gum, carrageenan, alginate, pectin, poloxamer, and mixtures thereof can be mentioned. Among the non-mucoadhesive polymers, for example, polyvinyl alcohol can be mentioned. Among the chelating agents, for example, edetate disodium and cromoglycate disodium can be mentioned. Among the antioxidants, for example, sodium metabisulfite, sodium bisulfite, acetylcysteine, ascorbic acid, and mixtures thereof can be mentioned.

[0082] By adjusting the amount and ratio of polyol, fatty acid and one or more active agents, the solubility of fatty acid and active agent and the physical state of formulation and the release profile of active agent can be controlled.In some embodiments, polyol, fatty acid, active ingredient and other necessary ingredients are configured in such a ratio that formulation provides rapid onset of action within about 5 minutes, within about 10 minutes, within about 15 minutes, or within about 30 minutes.

[0083] The transdermal formulations disclosed herein can provide sustained or continuous release of agents (e.g., CD38 inhibitors, NO boosters, or S-nitrosothiol-containing molecules). In some embodiments, the formulations provide sustained release (transdermal delivery into blood circulation) of agents for a period of 30 minutes, 1 hour, 2 hours, 4 hours, 8 hours, 10 hours, 12 hours, 15 hours, 18 hours, 24 hours, about 2 days, about 3 days, about 5 days, or about 7 days. The release rate can also be controlled by selecting a suitable ratio of polyol solvent and fatty acid. In some embodiments, one, two, or three of the following parameters can be achieved for the formulation: (a) less than 15%, less than 20%, less than 25%, less than 30%, or less than 35% of the drug is delivered into the blood circulation within about 30 minutes, about 1 hour, about 2 hours, about 3 hours, about 4 hours, or about 5 hours; (b) about 25% to about 90%, about 30% to about 85%, about 35% to about 70%, about 40% to about 70%, about 50% to about 60%, about 35% to about 50%, about 40% to about 60%, or about 35% to about 80% of the agent is delivered into the blood circulation within about 6 hours, about 8 hours, about 10 hours, about 12 hours, or about 14 hours; and (c) greater than 60%, greater than 70%, or greater than 80% of the drug is delivered into the blood circulation in about 16 hours, about 18 hours, about 20 hours, about 22 hours, about 24 hours, about 36 hours, or about 48 hours.

[0084] In some embodiments, the active agent and carrier (e.g., polyol or fatty acid) in the formulation and their amounts are selected such that a window of therapeutic effect is maintained for about 30 minutes, about 1 hour, about 2 hours, about 4 hours, about 6 hours, about 8 hours, about 10 hours, about 12 hours, about 14 hours, about 24 hours, about 2 days, about 3 days, about 5 days, or about 7 days, and the plasma concentration of the active agent fluctuates by less than 5%, less than 10%, less than 15%, less than 20%, less than 25%, less than 30%, or less than 40% during such window. In some embodiments, the window begins within about 10 minutes, within about 20 minutes, within about 30 minutes, within about 1 hour, or within about 2 hours after the formulation is administered.

[0085] The formulation may include a second drug. Examples of the second drug include hypertensive agents, antibacterial agents, anti-inflammatory agents, analgesics, anesthetic agents, antihistamines, antiseptics, immunosuppressants, antihemorrhagic agents, vasodilators, wound healing agents, antibiofilm agents, and mixtures thereof. Alternatively, the second drug may be in a separate formulation and / or administered separately from the transdermal formulations described herein.

[0086] Examples of anti-inflammatory agents include nonsteroidal anti-inflammatory drugs (NSAIDs); propionic acid derivatives such as ibuprofen and naproxen; acetic acid derivatives such as indomethacin; enolic acid derivatives such as meloxicam and acetaminophen; methyl salicylate; monoglycol salicylate; aspirin; mefenamic acid; flufenamic acid; indomethacin; diclofenac; alclofenac; diclofenac sodium; ibuprofen; ketoprofen; naproxen; pranoprofen; fenoprofen; sulindac; fenclofenac; clidanac; flurbiprofen; fentiazac; bufexamac; piroxicam; phenylbutazone; oxyphenbutazone; clofezone; pentazocine; mepirizole; tiaramide hydrochloride;Steroids such as clobetasol propionate, betamethasone dipropionate, halbetasol proprionate, diflorasone diacetate, fluocinonide, halcinonide, amcinonide, desoximetasone, triamcinolone acetonide, mometasone furoate, fluticasone proprionate, betamethasone diproprionate, triamcinolone acetonide, fluticasone propionate, desonide, fluocinolone acetonide, hydrocortisone braylate vlaerate), prednicarbate, triamcinolone acetonide, fluocinolone acetonide, hydrocortisone and others known in the art, prednisolone, dexamethasone, fluocinolone acetonide, hydrocortisone acetate, prednisolone acetate, methylprednisolone, dexamethasone acetate, betamethasone, betamethasone valerate, flumethasone, fluorometholone, beclomethasone diproprionate, fluocinonide, topical corticosteroids including hydrocortisone, hydrocortisone-21-monoesters (e.g., hydrocortisone-21-acetate, hydrocortisone-21-butyrate, hydrocortisone-21-propionate, The corticosteroid may be one of the lower potency corticosteroids such as hydrocortisone-21-valerate, hydrocortisone-17,21-diesters (e.g., hydrocortisone-17,21-diacetate, hydrocortisone-17-acetate-21-butyrate, hydrocortisone-17,21-dibutyrate, alclometasone, dexamethasone, flumethasone, prednisolone, or methylprednisolone, or it may be a higher potency corticosteroid such as clobetasol propionate, betamethasone benzoate, betamethasone dipropionate, diflorasone diacetate, fluocinonide, mometasone furoate, triamcinolone acetonide, etc.;

[0087] In some embodiments, the formulation contains an antiviral agent, such as acyclovir, trifluridine, idoxuridine, penciclovir, famciclovir, cidofovir, ganciclovir, valacyclovir, podofiloxin, podophyllotoxin, ribavirin, abacavir, delavirdine, didanosine, efavirenz, lamivudine, nevirapine, stavudine, zalcitabine, zidovudine, amprenavir, indinavir, nelfinavir, ritonavir, saquinavir, amantadine, interferon, oseltamivir, ribavirin, rimantadine, zanamivir, and combinations thereof. Antiviral treatment can be used to treat both localized and systemic viral infections, such as Covid-19, herpes labialis, or genital herpes.

[0088] Examples of antibacterial agents include penicillins and related drugs, carbapenems, cephalosporins and related drugs, erythromycin, aminoglycosides, bacitracin, gramicidin, mupirocin, chloramphenicol, thiamphenicol, sodium fusidate, lincomycin, clindamycin, macrolides, novobiocin, polymyxins, rifamycins, spectinomycin, tetracyclines, vanomycin, teicoplanin, streptogramins, antifolates including sulfonamides, trimethoprim and combinations thereof, and synthetic antibacterial agents including pyrimethamine, nitrofurans, methenamine mandelate, and methenamine hippurate, nitroimidazoles, quinolones, fluoroquinolones, isoniazid, ethambutol, pyrazinamide, para-aminosalicylic acid (PA S), cycloserine, capreomycin, ethionamide, prothionamide, thiacetazone, viomycin, ebeminomycin, glycopeptides, glyclyclyclines, ketolides, oxazolidinones; imipenem, amikacin, netilmicin, fosfomycin, gentamicin, ceftriaxone, ziracin, linezolid, cinelucid, aztreonam, and metronidazole, epiroprim, sanfetrinem sodium, biapenem, dynemycin, cefluprenam, cefoselis, sanfetrinem celexetil, cefpirome, mersacidin, rifalazil, cosane, lenapenem, beneprim, sulopenem, ritipenam acoxil, cyclothialidine, micacocidin A, carumonam, cefozopran, and cefetamet pivoxil.

[0089] Examples of antihistamines include diphenhydramine hydrochloride, diphenhydramine salicylate, diphenhydramine, chlorpheniramine hydrochloride, chlorpheniramine maleate isothipendyl hydrochloride, tripelenamine hydrochloride, promethazine hydrochloride, methdilazine hydrochloride, etc. Examples of local anesthetics include dibucaine hydrochloride, dibucaine, lidocaine hydrochloride, benzocaine, p-butylaminobenzoic acid 2-(di-ethylamino)ethyl ester hydrochloride, procaine hydrochloride, tetracaine, tetracaine hydrochloride, chloroprocaine hydrochloride, oxyprocaine hydrochloride, mepivacaine, cocaine hydrochloride, piperocaine hydrochloride, dyclonine and dyclonine hydrochloride.

[0090] Examples of preservatives include alcohol, quaternary ammonium compounds, boric acid, chlorhexidine and chlorhexidine derivatives, iodine, phenols, terpenes, germicides, disinfectants including thimerosal, phenol, thymol, benzalkonium chloride, benzethonium chloride, chlorhexidine, povidone iodine, cetylpyridinium chloride, eugenol, and trimethylammonium bromide.

[0091] Examples of analgesics include alfentanil, benzocaine, buprenorphine, butorphanol, butabamben, capsaicin, clonidine, codeine, dibucaine, enkephalin, fentanyl, hydrocodone, hydromorphone, indomethacin, lidocaine, levorphanol, meperidine, methadone, morphine, nicomorphine, opium, oxybuprocaine, oxycodone, oxymorphone, pentazocine, pramoxine, proparacaine, propoxyphene, proxymethacaine, sufentanil, tetracaine, and tramadol.

[0092] Examples of anesthetics include alcohols such as phenol; benzyl benzoate; calamine; chloroxylenol; dyclonine; ketamine; menthol; pramoxine; resorcinol; troclosan; procaine drugs such as benzocaine, bupivacaine, and chloroprocaine; cinchocaine; cocaine; dexivacaine; diamocaine; dibucaine; etidocaine; hexylcaine; levobupivacaine; lidocaine; mepivacaine; oxethazaine; prilocaine; procaine; proparacaine; propoxycaine; and pyrrocaine. ; lysocane; rhodocaine; ropivacaine; tetracaine; and derivatives such as pharma- ceutical acceptable salts and esters including bupivacaine HCl, chloroprocaine HCl, diamocaine cyclamate, dibucaine HCl, dyclonine HCl, etidocaine HCl, levobupivacaine HCl, lidocaine HCl, mepivacaine HCl, pramoxine HCl, prilocaine HCl, procaine HCl, proparacaine HCl, propoxycaine HCl, ropivacaine HCl, and tetracaine HCl.

[0093] Examples of antihemorrhagic agents include thrombin, phytonadione, protamine sulfate, aminocaproic acid, tranexamic acid, carbazochrome, carbaxochrome sodium sulfanat, rutin, and hesperidin. Further examples of second or additional agents include chemotherapeutic agents (such as tyrosine kinase inhibitors, immune checkpoint inhibitors, VEGF inhibitors, etc.) and glucose-lowering agents (e.g., metformin).

[0094] Further examples of second agents include daunorubicin, daunomycin, dactinomycin, doxorubicin, epirubicin, idarubicin, esorubicin, bleomycin, mafosfamide, ifosfamide, cytosine arabinoside, bischloroethylnitrosourea, busulfan, mitomycin C, actinomycin D, mithramycin, prednisone, hydroxypregesterone, testosterone, tamoxifen, dacarbazine, procarbazine, hexamethylmelamine, pentamethylmelamine, mitoxantrone, amsacrine, chlorambucil, methylcyclohexylnitrosourea, nacarbazine ... These include nitrogen mustard, melphalan, cyclophosphamide, 6-mercaptopurine, 6-thioguanine, cytarabine, 5-azacytidine, hydroxyurea, deoxycoformycin, 4-hydroxyperoxycyclo-phosphoramide, 5-fluorouracil (5-FU), 5-fluorodeoxyuridine (5-FUdR), methotrexate (MTX), colchicine, taxol, vincristine, vinblastine, etoposide (VP-16), trimetrexate, irinotecan, topotecan, gemcitabine, teniposide, cisplatin, and diethylstilbestrol. In some embodiments, second agents that can be used in combination with the formulations disclosed herein or that can be incorporated into the same transdermal formulation include anti-inflammatory agents, analgesics, antibacterial agents, antifungal agents, antibiotics, vitamins, and antioxidants. In some embodiments, the second agent is selected from piperine, anthranilic acid, benzophenones, camphor derivatives, cinnamates (e.g., octyl methoxycinnamate), dibenzoylmethanes (e.g., butyl methoxydibenzoylmethane), p-aminobenzoic acid (PABA) and its derivatives, salicylates, and PDE5 inhibitors (e.g., sildenafil (Viagra), tadalafil (Cialis), vardenafil (Levitra), and avanafil (Stendra)).

[0095] Any of the second agents described herein can be incorporated into the same transdermal formulation. Alternatively, in some embodiments of any of the methods disclosed herein, the second agent can be administered separately from the transdermal formulation via any suitable route, including oral, transdermal and parenteral routes.

[0096] In some embodiments, the combination of active agents in a transdermal formulation produces synergistic therapeutic effects.For example, curcumin and PDE5 inhibitors, when incorporated into the same transdermal formulation or administered in combination sequentially, can result in early symptom recovery (fever, cough, sore throat, and shortness of breath), reduced exacerbations, and reduced red flag signs in patients with viral infections (e.g., COVID-19).

[0097] Transdermal Delivery Systems The kit or transdermal delivery system may contain any combination of the components described herein in sufficient amounts for at least one agent, and may further include instructions recorded in tangible form for use of the components. For some applications, one or more components may be provided in premeasured, single-use amounts in individual, typically disposable patches, tubes, or equivalent containers.

[0098] The formulations disclosed herein can be incorporated into a transdermal delivery system or kit and can be used as a patch, swab, aerosol, cream, sponge, sprayer, nebulizer, or via other suitable means. The transdermal delivery system can also include instructions for administration of the formulations and one or more treatment methods disclosed in this patent document. The liquid or semi-solid formulation can be applied directly to the skin, for example, using a swab or sponge. Alternatively, the transdermal delivery system can include a layer coated or impregnated with a liquid, semi-solid, or solid transdermal formulation. For example, a patch can have a layer impregnated with a liquid formulation or coated with a semi-solid or solid formulation. The transdermal delivery system can also include an adhesive member for attaching it to the skin.

[0099] The transdermal delivery system or kit can be in any suitable shape for application to a subject in need thereof. For example, the sponge loaded with the formulation disclosed herein can be shaped as a round, cylindrical, conical, flat, tubular, and other symmetrical or asymmetrical shape for insertion into a body cavity or attachment or application to a target location, and can include an applicator or applicator portion. The sponge can be made of a material that absorbs liquid by capillary action. Alternatively, the material can be hydrophilic or hygroscopic, or coated with a hydrophilic or hygroscopic layer that shows an affinity for aqueous solutions, especially moisture from the site or body cavity where the reservoir is placed. The sponge with absorbent properties can be made of natural or synthetic materials, including, for example, polyester, polyurethane, and vegetable cellulose.

[0100] In some embodiments, the formulation is incorporated into a liquid reservoir. The reservoir can be used independently, attached to the sponge, or partially or completely encapsulated in the sponge. Alternatively, the contents of the liquid reservoir can be loaded into the sponge for application after mixing with necessary treatments or additional agents. For formulations containing nitrite and a nitrite source that requires acid to generate NO, the acid can be added to the reservoir containing the nitrite before administration. Alternatively, a dual liquid reservoir system can be employed. For example, a pouch contains a thiol-containing molecule and a nitrite in a polyol solvent system, and another pouch contains an acid source. Of course, additional pouches can be used to separately encapsulate the thiol-containing molecule or the nitrite or the fatty acid. Before administration or upon contact with the skin, the contents of the pouches are mixed to initiate the reaction of the acid with the nitrite and the subsequent nitration of the thiol-containing molecule. The pouch for enclosing the NO precursor or acid source is generally a frangible or permeable container that does not contact each other or is separated by a non-permeable removable barrier before administration of the formulation.When administered, the acid and the NO precursor can permeate through their respective pouches and mix with each other under pressure from the user after removing the barrier.The acid and the NO precursor can also be mixed by simply breaking the pouch during or before administration.In a further exemplary embodiment, the acid and the NO precursor are mixed in the container before administration.The resulting mixture is absorbed by a cotton swab, sponge, or absorbent patch, and then applied to the skin.

[0101] In some embodiments, the pouch has a permeable or semi-permeable membrane surface that is optionally coated with adhesive to attach the membrane to the skin. Instead of adhesive coating, the pouch can also be attached to the skin by holding it to the skin and then covering it with an adhesive patch or encapsulation sheet. Commercially available pouch stock materials such as DuPont's SURLYN® can also be used for the liquid reservoir. Additional examples include co-extruded ethylene acrylic acid / low density polyethylene (EAA / LDPE) material, or BAREX® (acrylonitrile-methyl acrylate) from INEOS.

[0102] In some embodiments, where the NO precursor is a mixture of thio-containing molecules and nitrite, the formulation can be incorporated into a patch. A layer of the patch is impregnated with the NO precursor in a polyol and fatty acid solvent system, while an acid source is placed in another layer. The two layers do not contact each other until the patch is attached to the skin or before administration. The contents of different layers can be mixed by applying pressure to the patch. Alternatively, the patch can include an impermeable barrier between the two layers, and the barrier can be removed before administration to mix the nitrite and acid.

[0103] In some embodiments, the formulation is a solid that contains a thickening agent or solidifying material such as cocoa butter. In some embodiments, the formulation is a solid or semi-solid that contains petrolatum. The solid or semi-solid formulation can be applied to the skin or can dissolve when rubbed into the skin with pressure.

[0104] In some embodiments, the formulation is loaded into a nebulizer or atomizer, which delivers the agent in aerosol form to the nose, mouth, or lungs of a subject in need thereof. Carbon dioxide or other suitable gas can be used as a propellant.

[0105] The system or kit can include any number of additional reagents or substances that are useful for carrying out the method of the present invention. The kit or system of the present invention can be provided at any temperature. For example, for storage of kits that include specific S-nitrosothiol-containing molecules in liquid or gel, they can be provided and maintained at a suitable temperature or at about 0°C.

[0106] The kit or system can also include packaging material for holding the instruction manual and the container or combination of containers.Instructions, such as written instructions or videotaped demonstrations, that detail the use of the transdermal formulation to treat target diseases and conditions can be included in the kit or system.Typical packaging materials for such kits and systems include solid matrices (e.g., glass, plastic, paper, foil, etc.) that hold the components in any of a variety of configurations (e.g., pouches, tubes, etc.).

[0107] Such kits or systems may also include information such as scientific literature references, package insert materials, clinical trial results, and / or summaries thereof, that show or establish the activity and / or benefits of the composition and / or describe dosage, administration, side effects, drug interactions, or other information useful to health care providers. Such information may be based on the results of various studies, for example, studies using laboratory animals, including in vivo models, and studies based on human clinical trials. The kits or systems described herein may be provided, sold, and / or promoted to health care providers, including physicians, nurses, pharmacists, formulary offices, and the like. The kits may also be sold directly to consumers in some embodiments.

[0108] How to use Transdermal formulations deliver high levels of curcuminoids and / or other potent anti-inflammatory and / or antioxidant agents locally and / or systemically, thus providing rapid intervention for a variety of diseases and conditions.

[0109] A physiologically natural way to enhance endothelial production of NO and decrease ROS levels is by decreasing CD38 levels. High levels of CD38 create low levels of NAD+, which in turn creates i) mitochondrial dysfunction leading to overproduction of ROS; and ii) decreased activity of sirtuin proteins. Taken together, these CD38-induced consequences lead to reduced production of NO from the endothelium, and overproduction of ROS and peroxynitrite from activated macrophages and microglia, which promote endothelial dysfunction. Furthermore, there is an interaction between senescent cells and CD38, each promoting the accumulation of the other. The accumulation of senescent cells in the endothelium not only prevents repair of the vascular intima, but also promotes persistent endothelial dysfunction, including pathological vascular remodeling and a continued decrease in NO production.

[0110] The formulation for sustained transdermal delivery of active agents results in a reduction in CD38 ectoenzyme activity, such as NADase activity; thereby enhancing NAD+ levels and promoting the activity of NAD+-dependent sirtuins. In so doing, this approach provides both a new method of prevention and treatment for a variety of diseases, including those resulting from endothelial dysfunction following acute or chronic inflammatory injury, age-related physical and cognitive skill decline, age-related cardiac tissue changes, vascular hypertrophy, osteoarthritis, peripheral neuropathy and long-COVID. Furthermore, this approach overcomes the negative consequences of CD38 overproduction on the ability of stem cells to differentiate into mature cells, which is at least partially responsible for chronic inflammatory anemia and the ineffectiveness of intrinsic stem cells and stem cell therapy to repair damaged tissues.

[0111] Transdermal formulations can be administered by any suitable route to deliver active ingredients across the skin, mucosa or membrane of the subject's body.Non-limiting examples of suitable routes include, for example, topical (e.g., instillation and mucosal routes, including vaginal and rectal delivery), pulmonary (e.g., by inhalation or insufflation of powder or aerosol, including by nebulizer), intratracheal, intranasal, and epithelial routes.In some embodiments, transdermal formulations include one or both of curcumin and quercetin, and optionally one or more of polyphenols, flavonoids, stilbenoids, and secosteroids.

[0112] Without being limited to any particular theory, it is hypothesized that the transdermal formulation disclosed herein delivers active agent transdermally to a subject and enhances systemic or local NO level in the subject.In some embodiments, the formulation contains an effective amount of NO booster to increase systemic or local NO level in the subject.In some embodiments, the formulation contains an effective amount of NO precursor, and the method converts the NO precursor into, for example, an S-nitrosothiol-containing molecule, which releases NO transdermally to the subject.In some embodiments, the formulation can contain both NO booster and NO precursor.In some embodiments, the formulation can also contain a second agent as defined above.

[0113] The transdermal formulation achieves NO enhancement through pathways including upregulation of endothelial nitric oxide synthase (eNOS), enhancing the activity of eNOS, and reducing the level of ROS. For example, ROS scavenges NO, causing eNOS decoupling, resulting in the shutdown of eNOS-associated NO synthase, and instead leading to further production of ROS by eNOS. Meanwhile, loss of endothelium results in i) loss of flow-mediated mechanotransduction mechanisms to control NO production from eNOS; and also results in loss of acellular areas next to the endothelium, which prevents NO scavenging by hemoglobin in red blood cells. The transdermal formulation of this patent document provides an effective amount of active agent that can enhance NO levels in the endothelial lining of blood vessels by inhibiting ROS from scavenging NO and limiting the degradation of the glycocalyx lining of the endothelium.

[0114] The transdermal formulation can be applied to a body surface or cavity of a subject. For example, a method of enhancing systemic or local NO levels or treating a disease or condition can include inserting a sponge loaded with the formulation disclosed herein between the cheek and gums.

[0115] In diseases or conditions associated with elevated CD38 levels, the transdermal formulations disclosed herein can reduce CD38 levels / activity and restore nicotinamide adenine dinucleotide (NAD), thus preventing and reversing many of those consequences of CD38 enhancement. NAD+ (NAD) is a key coenzyme found in all cells in the body, involved in hundreds of metabolic processes such as cellular energy and mitochondrial health. It is required for the activity of SIRT1 and SIRT3, which are essential for controlling inflammation, oxidative stress and cellular repair. CD38 is an enzyme found primarily, but not exclusively, on the surface of macrophages and microglial cells. CD38 has NADase activity (degrades NAD). The amount of CD38 on macrophages and microglial cells increases with age and the onset of acute and chronic inflammation, and as a result, NAD levels decline, resulting in decreased energy, fatigue, increased inflammatory processes, and an inability to repair damage to cellular components, including DNA. The transdermal formulations disclosed herein can effectively reduce CD38 levels with little or no side effects, which may help prevent and limit severe endothelial dysfunction following acute inflammatory insults, limit age-related physical and cognitive skill decline, reverse age-related cardiac tissue changes, and stabilize red blood cells.

[0116] Certain flavonoids, such as apeginin and quercetin, have additional advantages over curcumin due to their senolytic activity, in addition to many of the anti-inflammatory, anti-oxidative stress properties that are similar to curcumin. Senescent cells induce an increase in CD38 levels. By removing senescent cells, CD38 levels can be reduced, thus maintaining or restoring NAD levels. Furthermore, CD38 can be reduced when activated M1 macrophages are repolarized to M2 macrophages. Curcumin and flavonoids have the ability to repolarize macrophages. Uptake of these actives by circulating M1 macrophages via the transdermal / transmucosal route is proposed to be a mechanism for the rapid reduction of CD38 activity. CD38 inhibitors can be agents that repolarize macrophages, which then results in a reduction in CD38 activity.

[0117] A disease or condition associated with elevated CD38 levels refers to a disease or condition in which a subject is detected to have a level of CD38 higher than normal.Normal levels of CD38 can be easily obtained from healthy individuals using well-known procedures and statistically acceptable analyses.

[0118] Various diseases or conditions can be treated with the transdermal formulations disclosed herein.Non-limiting examples of diseases and conditions include age-related physical and / or cognitive decline, lupus, rheumatoid arthritis, multiple sclerosis, leukemia and multiple myeloma, cardiovascular disease, neurodegenerative disease, COVID-19 symptoms, severe and persistent (long covid), diabetes, hypertension, neuropathic pain, osteoarthritis, anemia of chronic disease, ALS, Parkinkinson's disease, ischemia-reperfusion injury, hypoxia-reoxygenation injury, transfusion-induced injury, radiation-induced injury including dermatitis, and neuroinflammation.

[0119] Other diseases or conditions treatable with the formulations disclosed herein include muscle structure disorders, neuronal activation disorders, muscle fatigue disorders, muscle mass disorders, metabolic disorders, vascular diseases, ocular vascular diseases, muscular eye diseases, renal diseases, hypertension, inflammation, endothelial dysfunction, dermatological conditions, ophthalmological conditions, bacterial infections, viral infections, ischemia-reperfusion injury, hypoxia-reoxygenation injury, cytokine storm phenomenon, sickle cell disease, inflammatory consequences of acute sickle cell crisis and other hemoglobinopathies (including HbE / β thalassemia), Chagas' disease, type 2 diabetes, lupus, and transient inflammatory conditions including leaky gut syndrome and "brain fog" due to chemotherapy.

[0120] In some embodiments, the disease or condition is a muscle structural disorder selected from Bethlem myopathy, central core disease, congenital fiber type inequality, distal muscular dystrophy (MD), Duchenne and Becker MD, Emery-Dreifuss MD, facioscapulohumeral MD, hyaline body myopathy, limb-girdle MD, muscle sodium channelopathy, myotonic chondrodystrophy, myotonic dystrophy, muscle tubular myopathy, nemaline body disease, oculopharyngeal MD, or stress urinary incontinence; a neuronal activation disorder is selected from amyotrophic lateral sclerosis, Charcot-Marie-Tooth disease, Guillain-Barre syndrome, Lambert-Eaton syndrome, multiple sclerosis, myasthenia gravis, nerve lesions, peripheral neuropathy, spinal muscular atrophy, tardy ulnar nerve palsy, and toxic myoneuropathy; a muscle fatigue disorder is selected from chronic fatigue syndrome, diabetes mellitus, muscle mass disorders are selected from cachexia, cartilage degeneration, cerebral palsy, compartment syndrome, critical illness myopathy, inclusion body myositis, polymyositis, muscle atrophy (inactivity), sarcopenia, steroid myopathy, and systemic lupus erythematosus. the β-oxidation disorder is selected from systemic carnitine transporter, carnitine palmitoyltransferase (CPT) II deficiency, very long-chain acyl-CoA dehydrogenase (LCHAD or VLCAD) deficiency, triglyceride deficiency, medium-chain acyl-CoA dehydrogenase (MCAD) deficiency, short-chain acyl-CoA dehydrogenase (SCAD) deficiency, and riboflavin-responsive disorder of β-oxidation (RR-MADD);Metabolic disorders include hyperlipidemia, dyslipidemia, hypercholesterolemia, hypertriglyceridemia, HDL hypocholesterolemia, LDL hypercholesterolemia and / or HLD noncholesterolemia, VLDL hyperproteinemia, dyslipoproteinemia, apolipoprotein AI hypoproteinemia, atherosclerosis, arteriosclerosis, cardiovascular disease, cerebrovascular disease, peripheral circulatory disease, metabolic syndrome, syndrome X, obesity, diabetes mellitus (Type I or II), hyperglycemia, insulin resistance, impaired glucose tolerance, hyperinsulinemia, diabetic complications, heart failure, myocardial infarction, cardiomyopathy, hypertension, nonalcoholic fatty liver disease (NAFLD), nonalcoholic steatohepatitis (NASH), blood clots, Alzheimer's disease, neurodegenerative diseases, demyelinating diseases, multiple sclerosis, adrenoleukodystrophy, dermatitis, psoriasis, acne, skin aging, hirsutism, inflammation, arthritis, asthma, irritable bowel syndrome, ulcerative colitis, Crohn's disease the cancer is selected from colon cancer, colorectal cancer, skin cancer, breast cancer, prostate cancer, ovarian cancer, and lung cancer; the vascular disease is selected from peripheral vascular insufficiency, peripheral vascular disease, intermittent claudication, peripheral vascular disease (PVD), peripheral arterial disease (PAD), peripheral arterial occlusive disease (PAOD), and peripheral occlusive arterial disease; the ocular vascular disease is selected from age-related macular degeneration (AMD), Stargardt's disease, hypertensive retinopathy, diabetic retinopathy, retinopathy, macular vascular disease, and degeneration, retinal hemorrhage, and glaucoma; the muscular eye disease is selected from strabismus, progressive external ophthalmoplegia, esotropia, exotropia, refractive and accommodative disorders, hyperopia, myopia, astigmatism, anisometropia, presbyopia, accommodative disorders, and internal ophthalmoplegia; the renal disease is selected from glomerulonephritis, glomerulosclerosis, nephrotic syndrome, hypertensive nephrosclerosis, acute nephritis, recurrent hematuria, persistent hematuria, chronic nephritis, rapidly progressive nephritis, acute renal failure, chronic renal failure, diabetic nephropathy, and Bartter's syndrome;

[0121] In some embodiments, the disease or condition is selected from hereditary lipodystrophy, non-alcoholic fatty liver disease (NAFLD), non-alcoholic steatohepatitis (NASH), renal ischemia / reperfusion injury (IRI), cardiac ischemia / reperfusion injury, Duchenne and Becker muscular dystrophy, diabetes (type I or type II), obesity, and sarcopenia. The disease or condition is selected from hereditary lipodystrophy, non-alcoholic fatty liver disease (NAFLD), non-alcoholic steatohepatitis (NASH), renal ischemia / reperfusion injury (IRI), cardiac ischemia / reperfusion injury, Duchenne and Becker muscular dystrophy, diabetes (type I or type II), obesity, and sarcopenia.

[0122] In some embodiments, the disease or condition is selected from Alpers disease, CPEO - chronic progressive external ophthalmoplegia, Kearns-Sayre syndrome (KSS), Leber's hereditary optic neuropathy (LHON), MELAS - mitochondrial myopathy, encephalomyopathy, lactic acidosis, and stroke-like episodes, MERRF - myoclonic epilepsy and ragged-red fiber disease, NARP - neurogenic weakness, ataxia, and retinitis pigmentosa, Pearson syndrome, platinum-based chemotherapy-induced toxic hearing loss, Cockayne syndrome, xeroderma pigmentosum A, Wallerian degeneration, and HIV-induced lipodystrophy.

[0123] In some embodiments, the disease or condition is a neurodegenerative disease, including dementia, Alzheimer's disease (AD), Parkinson's disease.

[0124] In some embodiments, the disease or condition is a tumor selected from glioblastoma, lung cancer, colon cancer, liver cancer, breast cancer, gastric cancer, bladder cancer, and melanoma.

[0125] In some embodiments, the disease or condition is an autoimmune disease selected from diabetes, rheumatoid arthritis (RA), multiple sclerosis (MS), and systemic lupus erythematosus (SLE).

[0126] In some embodiments, the disease or condition is an inflammatory disease selected from asthma, chronic obstructive pulmonary disease (COPD), pneumonia, and non-alcoholic steatohepatitis (NASH).

[0127] In further exemplary embodiments, the transdermal formulations and systems can be used as transdermal therapies to prevent, manage and reverse clinical consequences of inflammatory diseases including diabetes, COVID-19 infection and sickle cell disease, provide local treatment of hypertension or local treatment of osteoarthritis, reverse the acute inflammatory cascade (cytokine storm), improve the safety and efficacy of transfused stored red blood cells, treat cerebral malaria or Chagas disease, or treat other early stage acute inflammatory diseases.

[0128] Phytochemicals (e.g., curcuminoids) have been shown to have antiviral activity. For example, it has been recently shown that the glycocalyx can prevent viral access to ACE2-binding receptors on endothelial cells, thus limiting uptake and replication. When glycocalyx is degraded by endothelial dysfunction, viral access to ACE2-binding sites increases. Curcumin and many of these other phytochemicals protect and preserve the glycocalyx by reducing ROS production and enhancing endothelial NO production (see below). These phytochemicals also reduce inflammation-induced injury resulting from diet and obesity by normalizing lipid and glucose metabolism, including insulin production and utilization. To treat inflammation caused by toxic chemicals and metals, curcumin and other phytochemicals can chelate and eliminate toxicants from the blood. They also limit the inflammatory response to inhaled toxicants, thus reducing the tendency for progression to ARDS. Phytochemicals stabilize red blood cells, thus minimizing toxicant-induced hemolysis, a potent trigger of inflammation.

[0129] The formulations disclosed herein can address proinflammatory injury, including acute inflammatory injury caused by certain viral infections (e.g., SARS CoV2, dengue, and influenza), obesity and glucose-induced inflammatory triggers, and inflammation caused by exposure to toxic metals and chemicals. For example, in patients with long COVID, adverse effects due to COVID that appear after appearing to recover from the primary infection include brain fog, fatigue, pain, clotting problems, myocarditis, edema, etc. Most of these long COVID symptoms may be due to a continuing imbalance between pro- and anti-inflammatory factors that promote the development and persistence of endothelial dysfunction. The formulations and methods disclosed herein can be applied to the treatment of these clinical symptoms of long COVID.

[0130] In some embodiments of the treatment method, the transdermal formulations disclosed herein are applied to treat diseases or conditions commonly associated with "cytokine storm", including, but not limited to: COVID-19 infection, sepsis, systemic inflammatory response syndrome (SIRS), cachexia, septic shock syndrome, traumatic brain injury (e.g., cerebral cytokine storm), graft-versus-host disease (GVHD), or as a result of treatment with activated immune cells, e.g., IL-2-activated T cells, anti-CD19 chimeric antigen receptor (CAR) T cells. In addition to the effects on endothelial function, an active agent such as curcumin at a sufficient concentration acts to efficiently block the binding of the spike protein on SARS CoV 2 to the ACE2 binding site on endothelial and pulmonary epithelial cells, thus inhibiting viral replication in vulnerable subjects.

[0131] In some embodiments of the treatment method disclosed herein, the transdermal formulation is administered to treat vascular leakage caused by disease or condition.Non-limiting exemplary diseases or conditions include vascular leakage syndrome, infectious disease, inflammatory disease, especially inflammation of the systemic vasculature including the blood-brain barrier caused by sepsis, lupus, irritable bowel disease, inflammatory bowel disease, and chemotherapy.Vascular leakage is characterized by hypotension, peripheral edema, and hypoalbuminemia.Vascular leakage can also be associated with diseases caused by pathogens, especially viruses and bacteria.

[0132] In some embodiments of the treatment method disclosed herein, the transdermal formulation is administered to treat or reduce the risk of cardiovascular disease associated with endothelial dysfunction. Endothelial cells are important components of blood vessels and play an important role in cardiovascular homeostasis by regulating blood fluidity and fibrinolysis, vascular tone, angiogenesis, monocyte / leukocyte adhesion, and platelet aggregation. The occurrence of endothelial dysfunction destroys endothelial barrier permeability, which is part of the inflammatory response in the development of cardiovascular disease. Non-limiting examples of cardiovascular disease include coronary artery disease (CAD), such as angina pectoris and myocardial infarction (commonly known as heart attack), stroke, heart failure, hypertensive heart disease, rheumatic heart disease, cardiomyopathy, abnormal heart rhythm, congenital heart disease, valvular heart disease, carditis, aortic aneurysm, peripheral artery disease, thromboembolic disease, and venous thrombosis.

[0133] In some embodiments, the amount / dosage of active agent(s) is selected and / or the administration schedule is configured such that the method increases or decreases the level of biomarkers associated with cardiovascular disease in a subject by at least about 5%, at least about 10%, at least about 15%, at least about 20%, at least about 30%, at least about 40%, at least about 50%, or at least about 60% compared to the control (without treatment with transdermal formulation) or the level before treatment with the transdermal formulation disclosed herein.Non-limiting examples of biomarkers associated with cardiovascular disease include white blood cell count (WBC), erythrocyte sedimentation rate (ESR), serum C-reactive protein (CRP), cardiac troponin, creatine kinase (CK), CK-MB, and myoglobin. In some embodiments, the subject prior to treatment has an abnormal level of one or more biomarkers associated with cardiovascular disease, wherein the abnormal level of the one or more biomarkers is at least about 5%, at least about 10%, at least about 15%, at least about 20%, at least about 30%, at least about 40%, at least about 50%, or at least about 60% higher or lower than a normal level or a level in a healthy subject.

[0134] Further examples of infectious diseases commonly associated with "cytokine storm" or vascular leakage include, but are not limited to, coronaviruses (COVs, including CoVid-19 / (SARS-CoV-2) coronavirus disease), malaria, avian influenza, smallpox, pandemic influenza, adult respiratory distress syndrome (ARDS), and severe acute respiratory syndrome (SARS). Certain infectious agents include, but are not limited to, Ebola, Marburg, Crimean-Congo hemorrhagic fever (CCHF), South American hemorrhagic fever, dengue, yellow fever, Rift Valley fever, Omsk hemorrhagic fever virus, Kyasanur Forest, Junin, Machupo, Sabia, Guanarito, Garissa, Ilexa, or Lassa fever virus. In some embodiments, the infectious disease is caused by a virus, bacteria, fungus, helminth, protozoan, or hemorrhagic infectious agent. In some embodiments, the infectious disease is caused by coronavirus (including covid-19) Arenaviridae, Filoviridae, Bunyaviridae, Flaviviridae, or Rhabdoviridae viruses. In some embodiments, the transdermal formulations and methods described herein can be applied to the treatment of septic shock syndrome, a chronic inflammatory response to infectious disease.

[0135] The methods disclosed herein can also be applied to the treatment of various types of pain, including, for example, neuropathic pain, surgery-related pain, trauma, periodontal or other dental procedure-related pain, as well as orthopedic or arthritis pain.For example, periodontal or other dental procedure-related pain can be treated with a sponge inserted into the subject's mouth at a suitable location, such as between the cheek and gums.The transdermal formulation can be administered before or after the onset of pain.For example, the formulation can be administered to the subject before a surgical procedure as a preventative method to reduce pain.

[0136] Conventional drugs for neuropathic pain have various levels of side effects.The transdermal formulation disclosed herein can be used alone or in combination with conventional drugs, including, for example, gabapentinoids, tricyclic antidepressants, and / or selective serotonin-norepinephrine reuptake inhibitors as first-line drugs, lidocaine, capsaicin, and / or tramadol as second-line drugs, and morphine, oxycodone, botulinum toxin A, and other opioids as third-line treatment.As a result, the transdermal formulation provides the advantage of reducing dependency on conventional analgesics and minimizing side effects.

[0137] Transdermal formulations can also be applied to postoperative pain management.For example, for individuals undergoing surgery, including bypass and thoracic surgery, coronary artery, inguinal hernia repair, and lower limb amputation, transdermal formulations can serve as an alternative therapeutic agent to treat these aforementioned pain conditions.

[0138] As mentioned above, the formulation can be administered in any suitable form for the method disclosed herein.In some embodiments of any formulation or method disclosed herein, the formulation is in semi-solid or solid form and is rubbed or spread on the skin or mucosal surface of the subject.In some embodiments, the patch is coated or impregnated with the formulation in liquid, semi-solid or solid form.In some embodiments, one or more of the NO booster, NO precursor and acid source are in a liquid reservoir before administration.In some embodiments, the formulation is administered via a sprayer or nebulizer.In some embodiments, the subject is a human.In some embodiments, the presence of symptoms, signs and / or risk factors of the disease or condition to be treated is determined before starting administration of the formulation.

[0139] The transdermal formulation of this patent document can be administered to activate the NAD-dependent deacetylase enzyme sirtuin-1 (SIRT1) in a subject. Thus, various diseases or conditions associated with dysfunctional SIRT1 can be treated. Sirtuins are a type of NAD+-dependent protein deacetylase enzyme that regulate a wide variety of cellular activities, promoting cell survival and extending lifespan in response to environmental stress. Sirtuins exert their effects by removing acetyl groups from specific target proteins in the presence of oxidized nicotinamide adenine dinucleotide (NAD+). For example, the yeast sirtuin enzyme Sir2 (silent information regulator 2), originally identified for its role in silencing DNA transcription, has also been shown to promote cell survival in response to calorie restriction. Similarly, in C. elegans, the sirtuin enzyme SIR-2.1 has been shown to extend lifespan. In mammalian cells, the sirtuin enzyme SIRT1 (homolog of yeast Sir2 and C. elegans SIR-2.1 enzymes) deacetylates the tumor suppressor p53 to promote cell survival. SIRT1 was reported to regulate various pathways, including, for example, the restoration of angiogenic function and the secretion of proangiogenic factors in endothelial progenitor cells. A seminal paper demonstrated that SIRT1 is involved in protection against excessive inflammation and oxidative stress by deacetylating NFκB and forkhead box O transcription factors. In addition, SIRT1 inhibits cellular senescence, promotes keratinocyte differentiation, and protects against UV-induced DNA damage. Several studies have also demonstrated that downregulated or dysfunctional SIRT1 is associated with various diseases, such as diabetic milieu, and that overexpression of SIRT1 improves glucose intolerance and insulin sensitivity, protecting against diabetes. Thus, sirtuins appear to be activated as part of a beneficial cellular response to stress, resulting in cell survival and lifespan extension.

[0140] Activators of sirtuins may therefore be beneficial in effecting fundamental cellular processes that protect cells from stress, prevent or treat various diseases or conditions, and extend healthy lifespan.

[0141] Transdermal delivery of active agents (e.g., NO enhancing and SIRT1 activating therapeutics) allows for easy combination with oral treatments targeting other relevant disease pathways that are not effectively addressed via transdermally delivered agents. The method includes administering a transdermal formulation disclosed herein to a subject in need thereof. In some embodiments, the formulation includes (a) a therapeutically effective amount of a SIRT1 activator; (b) a polyol solvent in an amount sufficient to dissolve the SIRT1 activator; and (c) a fatty acid. The SIRT1 activator can be one or more NO boosters described above. In some embodiments, the SIRT1 activator includes one or more of curcuminoids, berberine, quercetin, resveratrol, and fisetin. The amount of activator can be adjusted depending on the nature of the agent and the disease or condition being treated. In some embodiments, the activator ranges from about 0.05% to about 40% by weight in the formulation. In some embodiments, the formulation provides a sustained release of the activator over a period of about 15 hours.

[0142] Treatment of acute and chronic diseases or other conditions may benefit from enhancing systemic nitric oxide levels in the endothelium and / or activating the SIRT1 and NRF2 signaling pathways. Non-limiting diseases or conditions include sickle cell disease, HbE / β thalassemia and other thalassemias, diabetic retinopathy, glaucoma, dry eye syndrome, and surgery-induced inflammation.

[0143] In some embodiments, the method enhances SIRT1 activity in a subject. The scope and composition of the formulation are as described above. In some embodiments, the formulation comprises (a) a therapeutically effective amount of a SIRT1 activator; (b) a polyol solvent in an amount sufficient to dissolve the SIRT1 activator; and, optionally, (c) a fatty acid. The SIRT1 activator can be one or more NO boosters described above. In some embodiments, the SIRT1 activator comprises one or more of curcuminoids, berberine, quercetin, resveratrol, and fisetin. The amount of activator can be adjusted depending on the nature of the agent and the disease or condition being treated. In some embodiments, the activator ranges from about 0.05% to about 40% by weight in the formulation. In some embodiments, the formulation provides sustained release of the activator over a period of about 1, about 2, about 4, about 8, about 10, about 15, or about 24 hours. In some embodiments, the disease or condition is selected from aging, chronic and acute inflammatory conditions, chemically induced vascular inflammation, viral infections, bacterial infections, and fungal infections. In some embodiments, the subject is diagnosed with a disease or condition selected from the group consisting of a neurodegenerative disease, diabetic kidney disease, diabetes, cardiovascular disease, endothelial dysfunction, muscular dystrophy, pain, neuroinflammatory conditions, abnormal vascular homeostasis, and lupus.

[0144] The transdermal formulations of this patent document can be administered to promote therapeutic effects or reduce adverse events of another therapy. In some embodiments of any method disclosed herein, the transdermal formulations of this patent document can be administered before, simultaneously with, or after another therapy, including, for example, orally administered drug therapy, intravenous infusion, intramuscular infusion, local medical treatment, and / or surgery. In some embodiments, the transdermal formulation is administered before additional therapy for a disease or condition. For example, local pretreatment with the formulations disclosed herein before transfusion can maximize tissue perfusion and minimize transfusion-related inflammation. Local pretreatment with the formulation or coadministration with another therapy can also reduce adverse events associated with the treatment (e.g., side effects associated with glucose-lowering drugs such as metformin, skin rash, chemotherapy-related oral mucositis / stomatitis).

[0145] The transdermal formulation can also enhance endothelial function in subjects.Therefore, it can treat various diseases or conditions related to dysfunctional or unbalanced endothelial function.Endothelium has two interrelated major elements that are essential for vascular homeostasis: glycocalyx and endothelial nitric oxide synthase (eNOS). Hair-like projections from the endothelium, called the glycocalyx, are involved in: i) maintaining vascular integrity, thus limiting vascular leakage and access of macrophages and lipids to the deeper layers of the vessel wall (triggering plaque formation); ii) controlling the overproduction of reactive oxygen species (ROS) by acting as a depot for the potent antioxidant superoxide dismutase (SOD); iii) regulation of blood flow in response to physiological demands through shear stress-regulated production of nitric oxide by eNOS; iv) restricting the access and binding of blood-borne cells (RBCs, monocytes, leukocytes), platelets, and infectious agents to the endothelium; v) limiting platelet activation; vi) Prevention of blood flow stagnation; vii) ensuring continued NO production by eNOS by preventing eNOS decoupling as a result of excess ROS. In a decoupled state, eNOS no longer produces NO, but instead produces ROS, leading to more inflammation; viii) maintaining a cell-free region along the endothelial layer, thus preventing scavenging of endothelium-generated NO by RBCs in close proximity to the endothelial layer; On the other hand, nitric oxide produced by endothelial nitric oxide synthase (eNOS) is essential for vascular homeostasis. Important functions of eNO include: i) Maintain tissue perfusion / oxygenation; ii) Prevent blood flow stagnation; iii) preventing a procoagulant environment; iv) repolarizing activated macrophages, thus promoting tissue repair and limiting tissue damage; v) regulating pro- and anti-inflammatory processes (balance between pro-inflammatory iNOS activity, which produces harmful peroxynitrite, and anti-inflammatory eNOS activity, which produces eNO, activation of SIRT1); vi) preventing inflammatory damage caused by ischemia-reperfusion and hypoxia-reoxygenation; vii) preventing ROS-induced damage, including lipid peroxidation and glycocalyx degradation; viii) Creating a depot of nitrosothiols stored within the endothelium and surrounding vascular lining that can rapidly provide NO under conditions where increased levels of NO are required, such as during extreme muscular activity.

[0146] Endothelial dysfunction is a physiological impairment of normal biochemical processes carried out by the endothelium, the cells that line the inner surface of blood vessels. Endothelial dysfunction is characterized by impaired endothelium-dependent vasodilation mediated by nitric oxide (NO) produced by endothelial nitric oxide synthase (eNOS), a constitutive form of NOS that is primarily expressed in endothelial cells. In healthy vasculature, NO produced by the endothelium diffuses to vascular smooth muscle cells (VSMCs), where it activates guanylate cyclase and stimulates the production of cyclic guanosine monophosphate (cGMP), thereby promoting VSMC relaxation and, consequently, vasodilation. Other functions of the endothelium (e.g., inhibition of platelet aggregation, inhibition of leukocyte adhesion, and inhibition of VSMC proliferation) are also mediated by NO. In dysfunctional endothelium, NO production is impaired. Endothelial dysfunction can be detected clinically, for example, by an elevated number of circulating endothelial cells (CECs).

[0147] Endothelial dysfunction is a common condition that is associated with, for example, hypertension, coronary artery disease, heart failure, stroke, peripheral artery disease, diabetes, chronic renal failure, abnormal vascular smooth muscle cell proliferation and other cardiovascular diseases, type 2 diabetes, insulin resistance and other metabolic syndromes, lupus, HIV, inflammation due to radiation and drug treatments (e.g., chemotherapy), hemoglobinopathies (sickle cell disease, HbE / β-thalassemia, cytokine storm-related conditions induced by viral diseases (e.g., SARS CoV, 2, dengue fever, influenza, hemorrhagic shock, hemorrhagic fever), erectile dysfunction secondary to surgery-induced inflammation, and inflammation associated with the increased population of senescent cells that typically occurs with aging. Furthermore, endothelial dysfunction is thought to be a key event in the development of atherosclerosis, predating clinically evident vascular pathology by many years. Endothelial dysfunction has also been shown to be of prognostically importance in predicting vascular events, including stroke and myocardial infarction. In addition, endothelial dysfunction has been shown to be involved in inflammation, infection, immune system dysfunction, sleep apnea, sepsis, chronic obstructive pulmonary disease, and exposure to proinflammatory agents.

[0148] The methods disclosed herein are applicable to the treatment of acute and chronic consequences of endothelial dysfunction.Examples of chronic consequences of endothelial dysfunction include the above-mentioned diseases and conditions.In some embodiments, the methods are applicable to the treatment of acute consequences, including, for example, cytokine storm and associated hypoxic / ischemic organ damage caused by physical activity or diet (e.g., heart attack caused by insufficient tissue perfusion / oxygenation), stroke, microemboli and macroemboli, pulmonary embolism, ischemia-reperfusion injury, hypoxia-reoxygenation injury, and long COVID, which is the result of ongoing chronic inflammation / endothelial dysfunction. In some embodiments, the method is applicable to treat chronic outcomes, including, for example, cardiovascular disease (CVD), coronary artery disease (CAD), renal failure, increased predisposition to cognitive decline and dementia, hypertension, sexual dysfunction, slow healing wounds, accelerated stent failure / closure, coronary artery bypass failure, slow healing wounds, reduced tolerance to physical activity due to mitochondrial dysfunction, accelerated age-related conditions, osteoarthritis, transient ischemic events, diabetic retinopathy, reduced insulin production due to inflammation-initiated damage to pancreatic β cells, HIV-induced CVD, CVS, and CAD secondary to ongoing periodic episodes of either sleep apnea and / or blood flow stagnation (e.g., sickle cell disease).Other applications of the method include blood transfusions and kidney dialysis, including either RBC or hemoglobin-based oxygen carriers (HBOC).

[0149] The transdermal formulations can be administered to treat a variety of local conditions associated with endothelial dysfunction. For example, local conditions such as slow-healing leg ulcers and erectile dysfunction are associated with underlying, often severe, endothelial dysfunction that limits blood flow to damaged tissues. Methods for treating slow-healing leg ulcers include sustained local delivery of nitric oxide to eliminate biofilms and infections that impede the therapeutic effects of agents designed to accelerate wound closure. In parallel, transcellular delivery of active agents such as curcumin normalizes the systemic vasculature, thus promoting tissue oxygenation and allowing stem cell migration and development. The transdermal formulations can be administered in combination with antibiotic agents or any suitable wound healing agent. In the case of erectile dysfunction, transdermal delivery of agents such as curcumin can be used alone or in combination with topical nitric oxide and / or oral PD5 inhibitors to restore systemic vascular health and reduce systemic inflammation. Given that systemic NO boosters (such as curcumin) enhance NO production in the endothelium and oral supplementation with PD5 inhibitors extends the duration of action of NO, the combination of a transdermally delivered active agent with an oral PD5 inhibitor would accelerate endothelial recovery for patients with endothelial dysfunction, including long-COVID and cytokine storm.

[0150] The transdermal formulations disclosed herein can also be administered to a subject in need thereof to reduce ROS production, peroxynitrite production (through inactivation of iNOS activity) and / or increase eNO production in the endothelium. Without being limited to any particular theory, it is hypothesized that the formulations provide pleiotropic effects, such as upregulation and / or activation of multiple anti-inflammatory and antioxidant enzymes, including sirtuin 1 (SIRT1) and other inflammation-regulating sirtuins, and signal transduction pathways; PPAR(γ) (peroxisome proliferator-activated receptor γ), peroxisome proliferator-activated receptor γ coactivator (PGC)-1α, is a member of a family of transcriptional coactivators that play a central role in regulating cellular energy metabolism. AMP-activated protein kinase (AMPK) is a phylogenetically conserved fuel-sensing enzyme present in all mammalian cells. When activated, AMPK stimulates energy-generating processes such as glucose uptake and fatty acid oxidation and reduces energy-consuming processes such as protein and lipid synthesis. The transcription factor Nrf2 (nuclear factor erythroid 2-related factor 2), a key regulator of antioxidant and cytoprotective genes, is mainly activated in response to oxidative stress. It is involved in the downregulation or inhibition of the SIRT1 / PGC-1α / PPAR-γ pathway, eNOS-mediated enhancement of eNO production, PPARP, Nrf2, heme oxygenase, AMPK, and ACE2 (angiotensin-converting enzyme 2, or ACE2 "receptor", protein, which provides the entry point for coronaviruses to hook and infect a wide range of human cells. The formulation can also be applied to downregulate or inhibit TLR4 (toll-like receptor 4 part of the mechanism of induction of inflammation), NADPH oxidase (NADPH oxidase (nicotinamide adenine dinucleotide phosphate oxidase) is a membrane-bound enzyme complex that faces the extracellular space and generates reactive oxygen species), and ACE (angiotensin I-converting enzyme).

[0151] The transdermal formulations described herein can be applied to the treatment and management of acute inflammation-induced injury and chronic consequences of many inflammation-induced conditions that may promote or be related to endothelial dysfunction (ED).Non-limiting examples of diseases or conditions related to endothelial dysfunction (ED) include cardiovascular disease, renal failure, cognitive decline, slow-healing wounds, hypertension, stroke, microembolism, edema, sexual dysfunction, retinopathy, neuropathy, and neuropathic pain.

[0152] For acute diseases or conditions, the formulation can rapidly initiate overall anti-inflammatory and antioxidant activity to impede and limit progression leading to the severe consequences of extreme ED. For example, a suitable patch or sponge loaded with the formulation provides an extremely high concentration of NO stimulator, such as curcumin, and is inserted between the gums and cheek for a period of time to ensure rapid and sustained delivery of therapeutic levels of curcuminoids or other anti-inflammatory, antioxidant and NO stimulators. This safe approach eliminates the concern of directly overdosing on NO, yet activates a full repertoire of host-based anti-inflammatory and antioxidant pathways. The use of systemically or locally delivered active agents, such as curcuminoids, can stimulate NO production in the vasculature and / or reduce the overproduction of reactive oxygen species (ROS). This combination of enhancing endothelially generated NO and halting ROS production is designed to prevent, limit and reverse ED and its consequences.

[0153] The transdermal formulations of this patent document can be administered to subjects in need thereof to treat diabetes and associated inflammation and other conditions. Inflammation of adipose tissue promotes insulin resistance and hyperglycemia, both of which cause and amplify endothelial dysfunction. Furthermore, chronic untreated endothelial dysfunction, which is generated and enhanced by hyperglycemia, excessive ROS production and other diabetes-related factors, is a common pathway by which type 2 diabetes and other inflammation-induced causes end-stage clinical conditions, including, for example, cardiovascular disease, renal failure, hypertension, stroke and microembolism, slow-healing wounds, sexual and bladder dysfunction, neuropathic pain, and cognitive decline. By lowering blood glucose, reversing insulin resistance, lowering the exacerbated and persistent blood glucose elevation induced by elevated blood glucose or surgery, lowering ROS levels, increasing NO levels in endothelium, and restoring vascular homeostasis, the transdermal formulations of this patent document minimize the negative and / or undesirable consequences associated with surgery, blood transfusion, stent implants, dialysis, and any other invasive procedures that may promote systemic inflammation. The formulation can upregulate Nrf2 and related antioxidant enzymes, including the potent antioxidant heme oxygenase (HO-1), and reduce oxidative stress (thereby allowing the recovery of damaged endothelium, treating endothelial dysfunction, and also preventing the onset or progression of endothelial dysfunction). In addition, it can limit the diabetes-augmented glucose response after surgery and shorten the recovery time of elevated glucose levels. Furthermore, it can limit stress-induced glucose spikes in stressed (e.g., injury, surgery, and blood transfusion) diabetic patients, and inflammatory consequences of stress, including urogenital dysfunction after radical prostatectomy.

[0154] Thus, the transdermal formulation can be administered to limit the negative clinical outcomes associated with inflammatory triggers in subjects who have pre-existing conditions or are at risk of developing conditions that promote hyperglycemia and subsequent endothelial dysfunction.In some embodiments, the formulation is administered prophylactically to subjects who are at risk of developing hyperglycemia.In some embodiments, the subject is determined to have hyperglycemia.

[0155] In some embodiments, the amount / dosage of the active agent(s) is selected and / or the administration schedule is configured such that the method restores insulin sensitivity and / or reduces elevated blood glucose by at least about 5%, at least about 10%, at least about 15%, at least about 20%, at least about 30%, at least about 40%, at least about 50%, or at least about 60% compared to the control (no treatment with a transdermal formulation) or the level before treatment with a transdermal formulation disclosed herein. In some embodiments, the subject is diagnosed with diabetes (e.g., type 2 diabetes) and has undergone invasive procedures such as surgery, blood transfusion, stent implant, and dialysis, or has undergone injury. In some embodiments, the subject before treatment has an abnormal level of blood glucose, the abnormal level being at least about 5%, at least about 10%, at least about 15%, at least about 20%, at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least 80%, or at least 100% higher or lower than the normal level or the level of a healthy subject.

[0156] The transdermal formulation disclosed herein can reduce the level of inflammatory cytokines.In some embodiments of the method disclosed herein, the amount / dosage of active agent(s) is selected and / or the administration schedule is configured so that one or more of biomarkers are reduced or modified by at least about 5%, at least about 10%, at least about 15%, at least about 20%, at least about 30%, at least about 40%, at least about 50%, at least about 60%, or at least about 100% compared to the control or the level before treatment with the transdermal formulation. Non-limiting examples of markers that may be reduced or altered by the transdermal formulations disclosed herein include TNF-α, TGFβ, MCP-1, IL-1α, IL-1β, IL-6, IL-10, IL-1, IL-18, MIF, TNF-β, MMP9, HIF-1, GLUT1, Hemox, PDK1, VEGF, CD11, EMR1, CXCR4, CCR5, IL-8, receptor for advanced glycation end products (RAGE), hsCRP, total antioxidant capacity (TAC), prostaglandins, leukotrienes, substance P, phosphatidylserine surface presentation on RBCs, select These include IL-1a / b, laminin and cahedrin, immunoglobulin receptors, chondroitin sulfate, syndecan-1, IL-1a / b, TNF-a, IL-6, D-dimer, and other markers reflecting a propensity for abnormal blood clotting, embolism, and thrombosis, such as C-reactive protein (CRP), Nrf2, NFκB, glutathione peroxidase (GPx), superoxide dismutase (SOD), syndecan-1, HMW-hyaluronic acid (1,000-6,000 kDa), A disintegrin and metalloprotease with thrombospondin type 1 repeats-13, protein C, von Willebrand factor, chondroitin sulfate, and sP-selectin. Additional examples include markers related to blood pressure, vasodilation, lung flow dynamics, vascular leakage / edema, M1 / M2 macrophage polarization, soluble platelet selectin, heparan sulfate, and cell and tissue oxygenation.In some embodiments, the subject prior to treatment has abnormal levels of one or more biomarkers or inflammatory cytokines, where the abnormal levels of the biomarkers or one or more cytokines are at least about 5%, at least about 10%, at least about 15%, at least about 20%, at least about 30%, at least about 40%, at least about 50%, at least about 60%, or at least about 100% higher or lower than normal levels or levels in a healthy subject.

[0157] In some embodiments of the methods disclosed herein, the subject has been diagnosed with a disease or condition selected from the group consisting of aging, chronic and acute inflammatory conditions, chemically induced vascular and / or pulmonary inflammation, viral infections, bacterial infections, and fungal infections.

[0158] Further examples of diseases or conditions that may be treated with the methods disclosed herein include neurodegenerative diseases, diabetic kidney disease, diabetes, cardiovascular disease, endothelial dysfunction, muscular dystrophies, pain, neuroinflammatory conditions, abnormal vascular homeostasis, lupus, Parkinson's disease, Alzheimer's disease, Huntington's disease, amyotrophic lateral sclerosis, neurodegenerative consequences of traumatic brain injury or cerebral hemorrhage, hypertension, inflammation, osteoarthritis, rheumatoid arthritis, endothelial dysfunction, dermatological conditions, ophthalmological conditions, bacterial infections, viral infections, ischemia-reperfusion injury, hypoxia-reoxygenation injury, cytokine storm phenomenon, cerebral malaria, Chagas disease, abnormal hemoglobinuria, and the like. In some embodiments, the conditions may include, but are not limited to, rhinitis, type 2 diabetes, coronaviruses, skin / dermatological conditions, acne, inflammatory skin conditions, Raynaud's disease, post-herpetic lesions, shingles, skin infections, wounds, burns, leg ulcers, sickle cell, diabetic, onychomycosis, peripheral vascular disease, infected and / or inflamed mucosal tissue, erectile dysfunction, female sexual dysfunction, vaginal infection / inflammation, catheter-associated urinary tract infections, sinusitis, cystic fibrosis, acute respiratory distress syndrome, pulmonary fibrosis, chronic obstructive pulmonary disease (COPD), bronchiectasis, lung infections including tuberculosis, pulmonary hypertension, as well as burns and other open wounds, inner ear infections, outer ear infections, gastrointestinal disorders, and acute vascular inflammatory conditions. Further examples of diseases treatable by the methods described herein include infectious diseases selected from the group consisting of coronaviruses (including SARS-CoV-2), Ebola, dengue fever, hemorrhagic shock, endotoxic shock, hemolysis and / or acellular hemoglobin toxicity resulting from the use of acellular hemoglobin-based blood substitutes (HBOC), Rift Valley fever, Marburg, Crimean-Congo hemorrhagic fever (CCHF), South American hemorrhagic fever, dengue, yellow fever, Omsk hemorrhagic fever virus, Kyasanur Forest, Junin, Machupo, Sabia, Guanarito, Garissa, Ilexa, and Lassa fever viruses.

[0159] Further examples of diseases or conditions treatable with the formulations disclosed herein include neurodegenerative diseases or disorders (e.g., Alzheimer's disease (AD), Huntington's disease, Parkinson's disease, amyotrophic lateral sclerosis (ALS), multiple sclerosis and disorders caused by polyglutamine aggregation); skeletal muscle diseases (e.g., Duchenne muscular dystrophy, skeletal muscle atrophy, Becker muscular dystrophy or myotonic dystrophy); metabolic disorders (e.g., insulin resistance, diabetes, obesity, impaired glucose tolerance, high blood cholesterol, hyperglycemia, dyslipidemia and hyperlipidemia); adult-onset diabetes, diabetic nephropathy, neuropathy (e.g., sensory neuropathy, autonomic neuropathy, motor neuropathy, retinopathy); bone disease (e.g., osteoporosis), blood disease (e.g., leukemia); liver disease (e.g., due to alcohol abuse or hepatitis); obesity; bone resorption, macular degeneration aging, AIDS-related dementia, AL S, Bell's palsy, atherosclerosis, heart disease (e.g., arrhythmias, chronic congestive heart failure, ischemic stroke, coronary artery disease, and cardiomyopathies), chronic degenerative diseases (e.g., myocardial disease), chronic renal failure, type 2 diabetes, ulcers, cataracts, presbyopia, glomerulonephritis, Guillain-Barre syndrome, hemorrhagic stroke, rheumatoid arthritis, inflammatory bowel disease, SLE, Crohn's disease, osteoarthritis, osteoporosis, chronic obstructive pulmonary disease (COPD), pneumonia, skin aging, urinary incontinence, mitochondrial dysfunction (e.g., mitochondrial myopathy, encephalopathy, Leber's disease, Leigh's encephalopathy, Pearson's disease, lactic acidosis, "mitochondrial encephalopathy, lactic acidosis, stroke-like symptoms" (MELAS), muscle diseases, including neuromuscular diseases such as muscular dystrophies and myopathies, and diseases or disorders associated with neuronal cell death, aging, or other conditions characterized by unwanted cell loss.In some embodiments, the disease or condition is Parkinson's disease, Alzheimer's disease, Huntington's disease, amyotrophic lateral sclerosis, and neurodegenerative consequences of traumatic brain injury or cerebral hemorrhage, sickle cell disease, thalassemia (e.g., HbE / beta thalassemia), diabetic retinopathy, glaucoma, dry eye syndrome, and inflammation caused by surgery, aging, chronic and acute inflammatory conditions, chemically induced vascular and / or pulmonary inflammation, viral infections, bacterial infections, fungal infections, diabetic kidney disease, diabetes, cardiovascular disease, endothelial dysfunction, muscular dystrophies, pain, neuroinflammatory conditions, abnormal vascular homeostasis, lupus, retinopathies including diabetic retinopathy, macular degeneration, peripheral vascular disease, long-term systemic consequences of chemotherapy and radiation therapy. , brain fog, rheumatoid arthritis, soft tissue injuries (muscle, tendons and ligaments), surgery-induced inflammatory sequelae (genitourinary dysfunction), transfusion-induced inflammation, inhibiting stent restenosis, limiting the inflammatory consequences of dialysis, inflammation and pain after dental treatment, neuropathic pain from any inflammation-induced injury including peripheral neuropathy, spinal neuropathy, arthritic pain, cognitive impairment in children due to cerebrovascular damage caused by sickle cell disease, cytokine storm due to coronavirus, dengue, Ebola, Rift Valley fever and influenza, cerebral malaria, metastatic spread of tumors through dysfunctional blood vessels, systemic consequences of psoriasis, dementia including Alzheimer's and Pick's disease, post-traumatic brain injury and hemorrhagic shock.

[0160] Autoimmune and immune-related disorders and diseases can also be treated or prevented with the methods described herein. Exemplary autoimmune and immune-related disorders include systemic lupus erythematosus, rheumatoid arthritis, osteoarthritis, juvenile chronic arthritis, spondyloarthropathies, systemic sclerosis, idiopathic inflammatory myopathy, Sjogren's syndrome, systemic vasculitis, sarcoidosis, autoimmune hemolytic anemia, autoimmune thrombocytopenia, thyroiditis, diabetes mellitus, immune-mediated renal disease, demyelinating diseases of the central or peripheral nervous system, idiopathic demyelinating polyneuropathy, Guillain-Barr syndrome, chronic inflammatory demyelinating polyneuropathy, and inflammatory inflammatory demyelinating polyneuropathy. , hepatobiliary diseases, infectious or autoimmune chronic active hepatitis, primary biliary cirrhosis, granulomatous hepatitis, sclerosing cholangitis, inflammatory bowel disease, gluten-sensitive enteropathy, Whipple's disease, autoimmune or immune-mediated skin diseases, bullous skin diseases, erythema multiforme, contact dermatitis, psoriasis, allergic diseases, asthma, allergic rhinitis, atopic dermatitis, food hypersensitivity, urticaria, immune diseases of the lungs, eosinophilic pneumonia, idiopathic pulmonary fibrosis, hypersensitivity pneumonitis, systemic lupus erythematosus, scleroderma, and arthritis.

[0161] Non-limiting examples of neurological diseases that may be treated or progression limited with the methods of this patent document include neurodegenerative disorders, such as stroke, Alzheimer's disease (AD), Parkinson's disease (PD), Huntington's disease (HD), amyotrophic lateral sclerosis (ALS; Lou Gehrig's disease), diffuse Lewy body disease, chorea-acanthocytosis, primary lateral sclerosis, multiple sclerosis (MS), and Friedreich's ataxia, periventricular leukomalacia (PVL), ALS-Parkinson's disease-dementia complex of Guam, Wilson's disease, cerebral palsy, progressive supranuclear palsy (Steel-Richardson syndrome), bulbar and pseudobulbar palsies, diabetic retinopathy, multi-infarct dementia, macular degeneration, Pick's disease, diffuse Lewy body disease, Creutzfeldt-Jakob, Gerstmann-Straussler-Scheinker, kuru, and fatal familial Prion diseases such as insomnia, primary lateral sclerosis, degenerative ataxias, Machado-Joseph disease / spinocerebellar ataxia type 3 and olivopontocerebellar degeneration, spinal and spinal-bulbar muscular atrophy (Kennedy disease), familial spastic paraplegia, Wolfert-Kugelberg-Welander disease, Tay-Sachs disease, multiple system degeneration (Shy-Drager syndrome), Gilles de la Tourette disease, familial dysautonomia (Riley-Day syndrome) Syndromes, Kugelberg-Welander disease, Subacute sclerosing panencephalitis, Werdnig-Hoffmann disease, Synucleinopathies (including multiple system atrophy), Sandhoff disease, Corticobasal degeneration, Spastic paraplegia, Primary progressive aphasia, Progressive multifocal leukoencephalopathy, Striatonigral degeneration, Familial seizure disorders, Chronic epileptic status associated with neurodegeneration, Binswanger disease, and Dementia (including all underlying etiologies of dementia).

[0162] Insulin resistance disorders treatable by the methods of this patent document include any disease or condition caused or contributed to by insulin resistance. Examples include diabetes, obesity, metabolic syndrome, insulin resistance syndrome, syndrome X, insulin resistance, high blood pressure, hypertension, high blood cholesterol, dyslipidemia, hyperlipidemia, dyslipidemia, stroke, coronary artery disease or atherosclerosis including myocardial infarction, hyperglycemia, hyperinsulinemia and / or hyperproinsulinemia, impaired glucose tolerance, delayed insulin release, diabetic complications including coronary heart disease, angina, congestive heart failure, stroke, cognitive function of dementia, retinopathy, peripheral neuropathy, etc. The prevention and treatment of disorders, nephropathy, glomerulonephritis, glomerulosclerosis, nephrotic syndrome, hypertensive nephrosclerosis, certain cancers (e.g., endometrial, breast, prostate, and colon), pregnancy complications, poor female reproductive health (e.g., menstrual irregularities, infertility, ovulation irregularities, polycystic ovarian syndrome (PCOS)), lipodystrophy, cholesterol-related disorders, e.g., gallstones, cholecystitis and cholelithiasis, gout, obstructive sleep apnea and respiratory problems, osteoarthritis, and bone loss, e.g., osteoporosis.Further application of the method of this patent document includes the promotion of wound healing, and can also be used to promote wound healing and diabetic disorder wound healing.

[0163] In some embodiments of any of the methods disclosed herein, the step of determining the subject as having downregulated or dysfunctional SIRT1 compared to a normal standard or reference is also included. In some embodiments, the methods disclosed herein further include a step of diagnosing the subject as having endothelial dysfunction or a disease or condition associated with endothelial dysfunction prior to administering the transdermal formulation to the subject.

[0164] Some embodiments of the methods disclosed herein include determining the subject as having systemic NO levels or plasma nitrite and / or nitrate levels that are at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 40%, at least 50%, or at least 60% lower than normal levels or a healthy reference standard.

[0165] In any of the methods disclosed herein, the treatment regimen can be administered after symptoms are clinically observed or after clinical symptoms of a disease or condition occur. Alternatively, the method can be used prophylactically before the onset or observation of any clinical symptoms. For example, the methods disclosed herein can address inflammation-induced injury, including acute inflammatory injury caused by certain viral infections (e.g., SARS CoV2, dengue fever, and influenza), obesity and glucose-induced inflammatory triggers, and inflammation caused by exposure to toxic metals and chemicals. Treatment can be administered when symptoms are identified or before the onset or observation of symptoms. In a further example, the transdermal formulation can be administered prophylactically to prevent postoperative inflammation-induced pain or inflammatory consequences of surgery, including erectile dysfunction.

[0166] The methods disclosed herein can increase NO levels systemically or locally. In some embodiments of any transdermal formulation or method of this patent document, the amount of active ingredient (e.g., NO booster and / or NO precursor, polyol, optional fatty acid, and / or other ingredient) in the transdermal formulation is selected so that it increases the subject's systemic or local NO level or plasma nitrite and / or nitrate level by at least 5%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 40%, at least 50%, at least 60% or more compared to the control or NO level or plasma nitrite and / or nitrate level before administration of the formulation. In some embodiments, the desired increase or change can be achieved within about 1 hour, within about 2 hours, within about 3 hours, within about 5 hours, or within about 8 hours. In some embodiments, the increase or change is maintained for a period of about 1 day, about 3 days, about 5 days, about 7 days, about 10 days, about 15 days, about 30 days or more. A variety of methods can be used to measure NO levels or plasma nitrite and / or nitrate levels, including colorimetric and chemiluminescent methods, for example using Griess reagent.

[0167] In some embodiments of any of the transdermal formulations or methods of this patent document, the amount of active ingredient (e.g., NO booster and / or NO precursor, polyol, optional fatty acid, and / or other ingredient) in the transdermal formulation is selected to reduce the subject's systolic and / or diastolic blood pressure and / or mean arterial pressure by at least 2, at least 4, at least 6, at least 8, at least 10, at least 12, at least 14, about 18, at least 20, at least 25, at least 30, at least 35, at least 40 mmHg or more over a period of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 days or more compared to a control or to the NO level or plasma nitrite and / or nitrate level prior to administration of the formulation.

[0168] In some embodiments of any method disclosed herein, the subject is diagnosed with hypertension or is at risk of developing hypertension.The transdermal formulation disclosed herein can be administered once, twice, three times, or as needed for 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 days or more.In some embodiments, the formulation, for example in the form of a patch, cream, or gel, is administered once every 1, 3, 5, 7, or 10 days.

[0169] Another aspect of the present disclosure provides a method for reducing CD38 activity or reducing elevated CD38 levels, comprising contacting a cell that overexpresses CD38 with a transdermal formulation as disclosed herein. The desired effect is best achieved through a combination of macrophage / monocyte / microglia repolarization from M1 to M2 phenotype, reduction of senescent cell population, and anti-inflammatory / antioxidant activity. In some embodiments, CD38 is present on the cell (on the surface of the cell or intracellularly). In some embodiments, the contact occurs in vitro. In some embodiments, the contact occurs in vivo.

[0170] Another aspect of the present disclosure provides a method for introducing a drug into a cell by contacting the cell with the formulation disclosed herein.As a result of the enhanced NO level, the method can stabilize the cell, improve storage properties, and reverse storage damage.In some embodiments, the cell is a red blood cell.In some embodiments, the formulation is in liquid form.In some embodiments, the formulation comprises an S-nitrosothiol-containing molecule.

[0171] Treating cells (e.g., red blood cells) with the formulations described herein can deliver NO boosters (e.g., curcumin) and optionally other anti-inflammatory / antioxidants into the lipid membrane or beyond the cytosol, stabilizing the cells for storage and converting the transfused cells into long-lasting (circulating) anti-inflammatory agents that can gradually deliver these agents to the endothelial lining of capillaries and other small-diameter vessels. Similarly, treating cells with formulations containing high concentrations of NO precursors, such as lipophilic S-nitrosothiols and S-NO derivatives of alkyl ester derivatives of NAC, will be effective in transnitrosating thiols within and on cells. Nitrosation of key thiols on and in red blood cells (including thiols of beta 93 and band 3 proteins on Hb) has been shown to stabilize red blood cells against oxidative damage, particulate formation, and hemolysis. Thus, such red blood cells can also be used as vehicles to deliver NO to thiols on the endothelium.

[0172] Another aspect provides a method for improving the safety and effectiveness of transfusion by contacting the cells or fluid to be transfused with the preparation before or during transfusion procedure.For example, the preparation containing NO booster or NO precursor can be administered before or during transfusion to protect red blood cells and enhance therapeutic effect.In some embodiments, the method includes contacting the preparation with cells or fluid 10 minutes, 15 minutes, 30 minutes, 1 hour, 2 hours, or 4 hours or more before transfusion is performed.

[0173] A related aspect provides a method for transdermally delivering an active agent to a subject in need thereof. The method comprises administering to the subject a transdermal formulation as disclosed herein. By delivering an effective amount of the active agent locally or systemically, a rapid and broad therapeutic effect can be achieved. The range of target diseases or conditions is as described above. In some embodiments, the transdermal formulation comprises one or more curcuminoids, and optionally one or more of polyphenols, flavonoids, stilbenoids, secosteroids, or natural products that promote NO production, as active ingredients for treating the disease or condition. In some embodiments, the transdermal formulation comprises one or both of curcumin and quercetin, and optionally one or more of polyphenols, flavonoids, stilbenoids, and secosteroids.

[0174] Manufacturing method Another aspect of the present disclosure provides a method for producing the formulation disclosed herein. The method generally includes preparing a solution of polyol solvent and fatty acid permeation enhancer, followed by adding an agent (NO booster or NO precursor). If necessary, the solution can be heated to a suitable temperature to dissolve the fatty acid and / or agent. In some embodiments, the active agent (e.g., NO booster or NO precursor or nitrite source) is saturated in the solution. After cooling the solution, the formulation can turn into a gel, and the precipitated excess active agent can be easily removed. As long as the NO booster or NO precursor is appropriately distributed in the formulation to achieve the desired therapeutic effect, variations in the conditions or order of mixing or adding different ingredients / components are also feasible.

[0175] In some exemplary embodiments, the drug dissolved in the solution is filled into a container (e.g., a sprayer or nebulizer; permeable or frangible pouch as described above) or soaked into a distribution vehicle (e.g., a cotton swab, a sponge, or the absorbent layer of a patch).When the NO precursor mixture contains a nitrite source, the acid source can be stored, for example, in a separate pouch or in a separate layer of the patch.The NO precursor mixture and the acid source can also be separated by a removable barrier disposed between two different compartments of the container or between two layers of the patch.

[0176] In some embodiments, the formulation is in the form of gel or semi-solid.The formulation can be adjusted by adding gelling agent or thickening agent.The formulation is then filled into a suitable container and then dispensed as gel, ointment, cream, emulsion, microemulsion, nanoemulsion, paste, balm, or other suitable form.The semi-solid formulation can also be coated on a backing material (e.g., the support layer of a patch).

[0177] Depending on the amount and nature of the thickener, the formulation can also be prepared in solid form. For example, a solution of NO booster (e.g., curcumin, demethoxycurcumin, bisdemethoxycurcumin, quercetin, berberine) can be mixed with molten pure cocoa butter and then cooled. The resulting solid formulation melts when rubbed into human skin with pressure.

[0178] General procedures for mixing reagents and handling manufacturing processes are well known to those skilled in the art through common knowledge or available in pharmaceutical technology handbooks, e.g., Remington: The Science and Practice of Pharmacy, 20th edition, Lippincott, Williams & Wilkins, Philadelphia, 2000, or the review article Souza et al, Topical ocular delivery of therapeutics: carrier systems and physical methods, J. Pharm. Pharmacol., 2013, 66, 507-530.

[0179] Dosing regimen The effective amount of the agent (NO booster or NO precursor) in the formulation or kit described herein to effectively enhance systemic NO levels depends on the route of administration, the type of subject, including humans, to be treated, and the physical characteristics of the particular subject under consideration. The dose or amount can be adjusted to achieve the desired effect, but will depend on factors such as body weight, diet, concomitant medication, and other factors that a person skilled in the medical field would recognize. More specifically, an effective amount or therapeutically effective amount refers to an amount of agent effective to increase systemic NO to a level that prevents, alleviates or improves symptoms of disease, or extends the survival of the subject being treated. The administration of the formulation can be adjusted to provide an optimal therapeutic response or long-term beneficial effect. For example, the formulation can be administered topically two or more times or three or more times per day. Alternatively, the amount or frequency of administration can be reduced as needed. The determination of the effective amount is well within the capabilities of one skilled in the art, especially in light of the detailed disclosure provided herein.

[0180] In non-human animal studies, application of the potential product is initiated at higher dosage levels, and the dosage is reduced until the desired effect is no longer achieved or adverse side effects disappear. Dosages can range widely, depending on the desired effect and therapeutic indication. Typically, the effective amount of the agent or dosage in the formulation can be from about 10 micrograms / kg to about 100 mg / kg body weight, preferably from about 100 micrograms / kg to about 10 mg / kg body weight. Alternatively, dosages can be calculated based on the surface area of ​​the patient, as will be understood by those skilled in the art.

[0181] In exemplary embodiments, the formulation is administered once daily, twice daily, three times daily, every other day, every third day, once weekly, once every two weeks, or once monthly.

[0182] The exact formulation, route of administration and dosage of a pharmaceutical composition can be chosen by the individual physician in view of the patient's condition (see, e.g., Fingl et al. 1975, "The Pharmacological Basis of Therapeutics", which is incorporated herein by reference in its entirety, see particularly page 1, chapter 1).

[0183] It should be noted that the attending physician will know how and when to terminate, interrupt, or adjust administration due to toxicity or organ dysfunction. Conversely, the attending physician will also know to adjust treatment to higher levels if the clinical response is not adequate (so as not to cause toxicity). The magnitude of the administered dose in the management of the disorder of interest will vary with the severity of the condition being treated and the route of administration. The severity of the condition can be evaluated, for example, in part, by standard prognostic evaluation methods. Furthermore, the dose and perhaps the dose frequency will also vary with the age, weight, and response of the individual patient. A program comparable to that discussed above can be used in veterinary medicine.

[0184] The formulations disclosed herein can be evaluated for efficacy and toxicity using known methods. For example, the toxicology of the formulation can be established by determining in vitro toxicity on cell lines, such as mammalian, preferably human cell lines. The results of such studies often predict toxicity in animals, such as mammals, and more specifically humans. Alternatively, toxicity in animal models, such as mice, rats, rabbits, or monkeys, can be determined using known methods. The efficacy of a particular compound can be established using several recognized methods, such as in vitro methods, animal models, or human clinical trials. Recognized in vitro models exist for nearly every class of condition. Similarly, acceptable animal models can be used to establish the efficacy of chemicals to treat such conditions. When selecting a model to determine efficacy, the skilled artisan can be guided by the state of the art to select the appropriate model, dose, and route of administration, and regimen. Of course, human clinical trials can also be used to determine the efficacy of transdermal f active agents in humans.

[0185] The formulation or kit described herein may be presented in a pack or dispenser device that can contain one or more unit dosage forms containing the formulation agent if desired. The pack may include, for example, metal or plastic foil, such as a blister pack. The pack or dispenser device may be accompanied by an administration instruction. The pack or dispenser may also include a notice associated with the container in a format prescribed by a government agency that regulates the manufacture, use, or sale of pharmaceuticals, which reflects the agency's approval of the drug form for human or veterinary administration. Such notice may be, for example, the label approved by the U.S. Food and Drug Administration for prescription drugs, or an approved product insert.

[0186] All references cited herein are incorporated by reference in their entirety. EXAMPLES

[0187] Example 1 Preparation of transdermal formulations Vehicles for loading the active ingredients were prepared as shown in Samples 1-4. Myristic acid (solid flakes) dissolved in the PEG400 upon warming the mixture in a warm water bath. Shaking accelerated the formation of the resulting clear solution. The following four concentrations of myristic acid (MA) in PEG400 were prepared and evaluated: Samples 1, 2, 3 and 4 were all clear solutions upon allowing the samples to reach ambient temperature. a. 30 ml PEG400 + 1.5 g MA (Sample 1) b. 30 ml PEG400 + 3 g MA (Sample 2) c. 30 ml PEG400 + 0.75 g MA (Sample 3) d. 30 ml PEG400 + 2.25 g MA (Sample 4)

[0188] Samples as vehicles were loaded with NO enhancing agents. Sample carrier formulations [or solvent systems] were loaded with 95% pure curcumin (Curcumin 95) as a NO boosting agent and tested for loading capacity. 1.5 grams of Curcumin 95 were added to Samples 1, 2, and 3. The resulting Curcumin 95 loaded samples were warmed and shaken until what appeared to be the maximum amount of curcumin dissolved, then cooled back to ambient temperature. All three samples allowed almost all of the added curcumin to dissolve, resulting in an intensely colored dark solution. Sample 3 offered higher solubility and remained liquid for several weeks. Samples 1 and 2 were initially liquid but formed a solid homogenous gel over a period of several hours. Sample 3 was prepared by dissolving 0.75 grams of MA in 30 ml of PEG400 and heating in a water bath (approximately 60-80 C) for approximately 15 minutes. Sample 4 was prepared by dissolving 2.25 grams of MA in 30 ml of PEG400 and heating in a water bath (approximately 60-80 C) for approximately 15 minutes. Both Samples 3 and 4 were homogeneous solutions at the end of the heating cycle. Upon cooling, Sample 3 remained liquid while Sample 4 formed a homogeneous gel.

[0189] Once loading capacity was established, sample transdermal formulations containing curcugen as an NO enhancer were made. Curcugen is a product containing curcumin, demethoxycurcumin (DMC) and bisdemethoxycurcumin (BDMC). The three curcuminoids account for approximately 50% by weight in curcugen. Sample formulations V3.3 and V4.3 were made by adding curcugen to Sample 3 and Sample 4 vehicles, respectively (3 grams of curcugen in 30 ml of either solvent). The resulting mixtures were heated and shaken for approximately 15 minutes, after which both appeared as homogenous solutions of dark maroon color. Upon cooling, V3.3 (3 grams of curcugen per 30 ml Sample 3 vehicle) remained liquid; whereas V4.3 (3 grams of curcugen per 30 ml Sample 4 vehicle) became a homogenous gel with the same dark maroon color (same color as V3.3 when heated).

[0190] No discoloration was observed over a period of at least three months for samples stored at ambient temperature. In contrast, when the same amount of curcumin was added to the same volume of water, the liquid phase was almost colorless and contained a significant amount of undissolved material.

[0191] V4.3 was loaded into the syringe upon warming and therefore in a liquid state. The liquid in the syringe gelled upon cooling to ambient temperature but could be squeezed out of the syringe as a gel that slowly dissolved on the skin. The applied gel was easily covered and captured with a waterproof and leak-proof transparent dressing (e.g. Mepitel transparent film bandage). The bandage prevented loss of the applied formulation and allowed it to remain in place for several days without loss of formulation. When the Mepitel bandage was removed several days later, none of the formulation was on the bandage; all of the formulation was within the skin as reflected by the absence of staining on tissue vigorously wiped over the pigmented skin that was under the bandage. The color was completely lost after 7-8 days.

[0192] The addition of water to PEG400 / MA-based formulations can create heterogeneous emulsions, making topical use difficult. The presence of water limits the solubility of curcumin, curcugen, quercetin, and berberine in PEG. For comparison, the same amount of curcumin was mixed with the same volume of water (i.e., 3 g of curcumin in 30 ml of water). The resulting mixture showed little color in the liquid phase and contained a significant amount of undissolved material.

[0193] The use of high concentration MA solutions with PEG as the base solvent facilitated the preparation of concentrated saturated solutions with poorly soluble NO enhancing actives such as curcumin, curcugen, quercetin, berberine and related molecules by simply adding an excess of the active agent to a heated high MA PEG / MA solvent system (e.g., V4.3) and allowing the mixture to cool. The resulting homogeneous gel was completely saturated with the active agent, while the undissolved excess remained solid at the bottom of the tube. The homogeneous saturated gel was then easily removed and separated from the undissolved material.

[0194] The Sample 3 solvent system was also tested as a vehicle for loading ability with the following poorly soluble agents using the method described above: quercetin, berberine, N-acetylcysteine ​​amide (NACA) and N-acetylcysteine ​​ethyl ester (NAC-ethyl ester). In all cases, the solvent system allowed for substantial dissolution of these difficult to solubilize drugs. Comparison of the aqueous solutions loaded with quercetin and berberine to the Sample 3 solutions revealed minimal dissolved material in the aqueous samples and a high degree of solubilization in Sample 3.

[0195] Similar results were obtained for quercetin in Samples 3 and 4 as vehicles, although the solubility of quercetin in these solvents was less than that of curcugen (approximately 200-300 mg quercetin in 30 ml of either Sample 3 or Sample 4). Similar to curcumin and curcugen, quercetin-loaded Samples 3 and 4 do not form gels and form gels, respectively, at ambient temperature.

[0196] The PEG400 / MA solution was combined with other delivery vehicles (e.g., petrolatum). Sample 3, containing curcumin, was mixed with petrolatum to produce a uniformly colored gel that remained stable (no fading) over a period of more than two months. Sample 3, which dissolved NAC amide and SNO derivatives of NAC, was easily combined with petrolatum to produce a stable peach-colored jelly. The stability is likely due to the low water activity and high viscosity, which inhibits loss of NO from the thiol groups. Different size aliquots of Sample 3 (containing both curcumin and curcugen) were mixed with molten pure cocoa butter and then cooled. The optimized mixture produced a solid, uniformly yellow / orange colored block / tube of cocoa butter that remained solid at ambient temperature. The solid material melted when rubbed into human skin with pressure. Similar results were obtained with coconut oil, but the low melting point of coconut oil made it difficult to apply to human skin without smudging or dripping. Combinations of the two oils were also tested. The use of PEG400 / MA with cocoa butter showed the most promising properties as a topical delivery vehicle suitable for cosmetic and dermatological applications due to a more favorable hardness for the resulting curcumin-loaded cocoa butter. Adding increasing amounts of PEG400 / MA solvent to the cocoa butter ultimately resulted in a gel-like material that remained color-wise stable.

[0197] Solutions of the two lipophilic NAC derivatives were prepared with sodium nitrite-saturated PEG400, resulting in clear solutions. The lipophilic derivatives of NAC dissolved readily under conditions in which NAC has limited solubility. NAC is water-soluble and the two derivatives are slightly soluble in water. The two solutions were then treated with a few drops of acetic acid to cause the formation of nitrous acid from the nitrite, which can then nitrosate the reactive thiols on both NAC derivatives. The solution turned pink, indicating the formation of S-nitrosothiols. The solution remained pink for several days. The corresponding aqueous solution lost its color within a few hours. The use of nitrite-saturated PEG400 allows for a formulation that can be used to generate NO and S-nitrosothiols by mixing with a reagent that acidifies the mixture. The dual frangible pouch incorporated into the patch can be used for a sustained NO delivery vehicle suitable for topical and transdermal applications.

[0198] Example 2 The effect of the V2.3 formulation (2 g curcugen per 30 ml of sample 3 vehicle) on blood pressure (BP) in rats was examined. The formulation was applied topically to the shaved abdomen of five Sprague-Dawley rats. A Q-tip saturated with V2.3 was rubbed into the shaved abdomen of the rats. In all cases (N=5), application of V2.3 resulted in a 20% reduction in systemic BP within 15-20 minutes of application. The reduction in BP was maintained for a 3-hour observation window without signs of recovery. Results from a second laboratory (N=6) in which six rats were similarly tested showed similar results (a 20% reduction in BP). Similar results to V3.3 were consistently obtained in one human subject when V3.3 was applied to the forearm (approximately 0.05 ml). Comparable BP reductions occurred within 15-20 minutes of application, and the reduced BP was sustained for several hours, gradually returning to the higher aforementioned initial values ​​after 12-24 hours. Similar results were obtained using V4.3 in the same human subjects. Cocoa butter doped with Sample 3 (V1.3) also showed similar physiological results when applied to one human test subject. Control Sample 3, which did not contain one or more curcuminoids or curcugen, did not induce any physiological results when applied to either rats or humans.

[0199] V3.3 was applied to a skin flap containing an optical window in a healthy hamster. The optical window allowed monitoring of blood vessels in the dermal layer beneath the site of V3.3 administration. A dose-dependent increase in blood vessel diameter at the site of topical application was observed within minutes of application.

[0200] Effects on NO plasma levels. Plasma levels of NO degradation products (nitrite / nitrate) were measured after topical application of V3.3 in four rats. Plasma levels of NO degradation products (nitrite / nitrate) in two of the rats tested were shown to increase by 15%, consistent with a reduction in blood pressure resulting from enhanced production of nitric oxide due to the known ability of curcumin to upregulate endothelial nitric oxide synthase (eNOS) and thus NO production. Two control animals showed no such increase under the same conditions. The results indicate that transdermal curcuminoids can increase systemic NO levels.

[0201] The drop in blood pressure within 15-20 minutes after topical application of the curcumin-containing sample indicates that curcumin is being delivered transdermally and that therapeutically effective levels are present within that short period of time. The drop in blood pressure and increase in nitrite / nitrate plasma levels are consistent with the known effect of curcumin on upregulating nitric oxide production in the endothelium via eNOS. The physiological response persists for several hours, consistent with locally delivered curcumin / curcgen being delivered into the circulation in a sustained manner. In contrast, curcumin delivered into the circulation via oral route or IV has a circulation time of only 2 hours due to the liver's rapid conversion of curcumin to an inactive drug. Even large doses of oral curcumin were not observed to result in this pronounced and prolonged physiological response.

[0202] Physiological Response in Animals Transdermal curcuminoid formulations V3.3 (curcugen 9g, myristic acid 2.25g, PEG400 90ml) and V4.3 (curcugen 9g, myristic acid 6.75g, PEG400 90ml) were also investigated.

[0203] Blood pressure was measured in rodents following topical application of transdermal curcuminoid formulations. BP was reduced by up to 20% in both rats and mice, with V3.3 being more effective than V4.3 in both the time it occurred and the extent of BP reduction.

[0204] It was also observed that topical application of either V3.3 or V4.3 resulted in an increase in plasma nitrite and nitrate levels over a 3-hour monitoring window following a single topical dose. Over the same period, BP underwent a sustained decrease (over a 3-hour window). After topical application of V3.3 to rats (N=3), a sustained accumulation of detectable concentrations of plasma curcuminoids was observed over a 3-hour window. Oral curcumin plasma levels peak at 1 hour and decline to undetectable levels within 3 hours.

[0205] Example 3 Control of systemic inflammation via transdermal delivery of NO boosters or NO precursors Inhibition of the development of severe vascular leakage in an acute inflammatory rat model was investigated using topically applied Vascarta formulation V3.3. The study used the following lipopolysaccharide (LPS)-induced cytokine storm protocol: a, LPS (10 mg / kg) was injected IP every 24 h to initiate and maintain the acute inflammatory response. b. LPS-treated rats were subjected to topical application of Vascarta formulation V3.3 (0.1 ml) starting from the time of the first LPS treatment and repeated every 24 hours for 3 days. c. Physiological parameters were measured from blood samples taken every 24 hours. d. Three days later, animals were anesthetized and surgically dissected to allow in vivo fluorescent imaging of the macro- and microvasculature. i. Formulations of fluorescently labeled albumin and dextran (500 kDa) were infused IV and fluorescence-derived images of the vasculature and surrounding tissue were used to determine the rates of extravasation out of the vasculature and into the surrounding tissue for both albumin and dextran.

[0206] Topical application of vehicle (PEG400 / MA) to LPS-treated rats revealed a pattern of rapid and extreme leakage for both albumin and the much larger dextran, consistent with that observed under conditions of severe acute inflammation. Daily topical application of V3.3, however, dramatically reduced the amount of leakage for albumin. The low levels of leakage observed are nearly identical to those observed for control animals. Similar results were observed for dextran. Extensive vascular leakage is a potentially fatal consequence of cytokine storms, regardless of cause (COVID-19, Ebola, Dengue, hemorrhagic shock, endotoxic shock, Rift Valley fever, etc.). The transdermal formulation provided dramatic positive intervention results with far-reaching clinical implications.

[0207] Example 4 This study evaluated the potential therapeutic efficacy of a transdermal delivery formulation according to the present invention (V4.3: 9g curcugen, 6.75g myristic acid, 90ml PEG400) in an acute vascular inflammation mouse model. Three cohorts, each with 3 subjects, were studied and compared. In cohort 1, subjects were injected with LPS but not topically treated (untreated). In cohort 2, subjects were injected with LPS topical treatment (0.1ml V4.3). In cohort 3, LPS injection was followed by topical treatment (0.1ml V4.3) 4 hours after LPS injection. Endotoxemia was induced by injection of 10mg / kg LPS (lipopolysaccharide from e. coli serotype 0128:B12, Sigma Aldrich St. Louis, MO). The procedure was the same as that described in a previously published study (Williams AT, Muller CR, Govender K, Navati MS, Friedman AJ, Friedman JM, Cabrales P. Control of systemic inflammation through early nitric oxide supplementation with nitric oxide releasing nanoparticles. Free Radic Biol Med. 2020;161:15-22. Epub 2020 / 10 / 05. doi: 10.1016 / j.freeradbiomed.2020.09.025. PubMed PMID: 33011274; PMCID: PMC7529593 and references therein).

[0208] Topically applied V4.3 (curcuminoid dissolved in PEG400 / myristic acid mix) was observed to act as both a preventive and interventional treatment for lipopolysaccharide (LPS)-induced cytokine storm. Results include vascular outcomes and cytokine profiles as a function of time for three different groups of LPS-treated mice: i) no topical treatment with V4.3; ii) pretreatment with topical V4.3 before LPS treatment, and iii) topical treatment with V4.3 after the onset of LPS-initiated cytokine storm. With regard to the response of the microvasculature to LPS treatment in the three groups, treatment with topical V4.3 limited arterial dilation, an indicator of shock-induced vascular collapse, and maintained arterial blood flow. It was also observed that topical V4.3 prevented the rapid fall in functional capillary density (FCD) that occurs in LPS-induced endotoxemia. V4.3 was effective as both a preventive and interventional treatment in preventing rapid fall in FCD compared to baseline (BL). FCD correlates with survival in that it reflects the ability to maintain tissue perfusion and deliver oxygen to tissues. Furthermore, both pretreatment and interventional treatment with locally applied V4.3 were found to limit the production of inflammatory cytokines. These results confirmed the efficacy of the transdermal formulation in limiting inflammatory outcomes when administered locally as a prophylactic agent or as an active therapeutic agent following the onset of inflammation.

[0209] Example 5 This study evaluated the prevention of LPS-induced vascular leakage via topically administered curcuminoids using a transdermal formulation according to the present invention in an LPS endotoxemia rat model. LPS (E. coli O26:B6) was inoculated into the rat model (10 mg / kg / day). Transdermal curcumin formulation (V3.3: 9 g curcugen, 2.25 g myristic acid, 90 ml PEG400) or vehicle control was applied daily for three days (0.1 ml / dose). On the third day, animals were surgically prepared for in vivo and fluorescence microscopy. Treatment with transdermal curcumin formulation was observed to prevent early-phase LPS-induced leakage (first 4 hours). Vascular leakage is dramatically reduced after topical application of V3.3 in an LPS-induced inflammatory mode. On the other hand, topical application of V4.3 (9 g curcugen, 6.75 g myristic acid, 90 ml PEG400) both before and after LPS-induced inflammation in mice (N=3) reduced the LPS-induced increase in inflammatory cytokine levels.

[0210] This experiment, combined with the results of previous studies, indicates that the topical transdermal curcumin formulation of this patent document can limit inflammation-induced vascular leakage, which is a hallmark of cytokine storm and other pro-inflammatory conditions.

[0211] Example 6 The skin permeability of the transdermal formulations according to the invention was studied. The skin permeability of the following transdermal formulations according to the invention was studied using confocal microscopy. The formulations differed only in the amount of myristic acid. a. Lot 43: V4.3 Curcugen 9g, Myristic Acid 6.75g, PEG400 90ml b. Lot 39: V3.3 Curcugen 9g, Myristic Acid 2.25g, PEG400 90ml c. Lot 38: V0.3 Curcugen 9g, PEG400 90ml

[0212] The formulation solidified as the temperature approached zero degrees Celsius. As the temperature was increased above ambient, the viscosity visibly decreased for all samples. The viscosities of the formulations and their components are as follows: water 0.9 cP; PEG400 (100%) 99 cP; PEG400 / water 90%, 80 cP; V3.3: 149.8 cP; PEG400 + myristic acid, V3.3: 149.8 cP; PEG400 + myristic acid, V4.3:4220 cP.

[0213] Curcuminoids have a broad absorbance spectrum with a maximum absorbance at 425 nm. The integrity of all human skin samples was measured using TEWL data, and all samples used had good integrity of the skin barrier. A DAPI filter was used to visualize the penetration of curcuminoids. All images were taken with the same settings.

[0214] All three formulations demonstrated penetration of curcuminoids into the skin. At the earliest time point of 1 hour, penetration of curcuminoids into the stratum corneum was observed with all three formulations.

[0215] In the lot 38 formulation, an expansion of the fluorescent band occurred between 3 and 6 hours in both donors. In the lot 39 and 43 formulations, no expansion of the band was observed up to 24 hours. This expansion suggests penetration of the curcuminoids into the upper epidermal layers just below the stratum corneum. In the lot 39 formulation, the amount of curcuminoids decreased in both donors between 3 and 6 hours. However, a decrease in intensity for the curcuminoids in the lot 38 and 43 formulations was observed after 6 hours. In the lot 38 formulation, a fluorescent spot was seen below the stratum corneum at early time points. However, no fluorescent spot was seen above the stratum corneum in the lot 39 and 43 formulations. Transepidermal water loss (TEWL) values ​​were comparable across all formulations and all time points, suggesting that the barrier integrity of all human skin samples was the same.

[0216] The decrease in fluorescence over time in the three formulations is an indication of the penetration of the curcuminoids into deeper layers of the skin. (Due to the high autofluorescence of untreated skin, it is possible that a lower amount of penetration of the active substance was not visualized and / or quantified). On this basis, the lot 39 formulation shows the fastest penetration of the curcuminoids into the skin. The depth of the fluorescence was also determined. This was measured at values ​​between 5 microns and 25 microns. The lot 38 formulation showed the fastest expansion of the fluorescent band, suggesting a faster skin penetration of the curcuminoids.

[0217] Example 7 The effect of the patented transdermal formulation on limiting or preventing the development of cardiovascular-induced inflammation in diabetic rat models was investigated in ZDSD diabetic rat models. Three rats were on a normal diet for 60 days without symptoms of diabetes. From about day 65, the rats were started on a high-fat diet, which led to a gradual rise in blood glucose. The animals were topically treated with formulation V4.3 every two days for the entire study period up to day 80. The cytokine profile showed the development of inflammation at about day 75. Treatment with formulation V4.3 limited the increase in inflammatory cytokines seen in sham at day 75. In particular, the profile of IL-18 showed clear evidence of the formulation in preventing the increase in IL-18, which is a marker for the tendency to develop cardiovascular outcomes in diabetic patients.

[0218] Example 7 The effect of treatment of mice with severe progressive endothelial dysfunction was investigated with formulation V4.3. A total of 24 male C57BL / 6J mice aged 6-8 weeks were used in the study. Mice were housed in an animal facility under a 12 / 12 h day / night cycle and free access to food and water. The study was carried out over a 4-week period. Week 1 was used for adaptation, as the animals were divided into experimental groups. Two groups received L-NAME (50 mg / kg) in the drinking water for 2 weeks (weeks 2 and 3) to chronically induce nitric oxide synthase (NOS) inhibition. One L-NAME-treated group received topical formulation V4.3 at a daily dose of 0.1 ml for the last 10 days before characterization. An L-NAME-untreated group was used as the sham group.

[0219] Topical treatment was initiated after the onset of pathology. Results showed direct evidence of treatment restoring elements of normal endothelial function (reduced oxidative stress in plasma and RBCs, reduced leukocyte adhesion to the endothelium indicating restoration of glycocalyx, and improved functional capillary density). Compared to the sham group, the group treated with the transdermal formulation showed lower levels of TNF-α, TGFβ, MCP-1, IL-1α, IL-1β, IL-6, IL-10, and IL-10. Better outcomes from the treated groups were also observed in terms of microhemodynamic changes, cell adhesion, vascular response of isolated aortic vessels, changes in erythrocytes and plasma antioxidants, hypoxia, reoxygenation, and systemic hemodynamic changes.

[0220] Table 1. Changes in body weight, relative tissue weight and water intake at the end of the study TIFF2025072314000001.tif41164

[0221] Table 2. Changes in red blood cell and plasma antioxidants at the end of the study TIFF2025072314000002.tif48144

[0222] It will be understood by those skilled in the art that the invention described herein is not limited to what has been particularly shown and described. Rather, the scope of the present invention is defined by the following claims. It should be further understood that the above description is merely representative of illustrative examples of embodiments. This description does not attempt to exhaustively list all possible variations. Alternative embodiments may not be presented for a particular component of a composition, or step of a method, and may result from a different combination of the components described, or other undescribed PEG alternative embodiments may be available for a formulation, kit, or method, and should not be considered as a disclaimer of those alternative embodiments. It will be understood that many of these undescribed embodiments are within the literal scope of the following claims, and others are equivalent.

Claims

1. below: (a) an effective amount of a NO booster comprising one or both of an effective amount of a curcuminoid and an effective amount of at least one flavonoid, wherein the curcuminoid is selected from the group consisting of curcumin, demethoxycurcumin, and bisdemethoxycurcumin, and the at least one flavonoid is selected from quercetin, apigenin, fisetin, luteolin, and rapamycin; (b) a polyol in an amount sufficient to dissolve the effective amount of curcuminoid and the at least one flavonoid; and optionally (c) a fatty acid, wherein the polyol and the fatty acid are in a ratio ranging from about 10:1 to about 50:1 by weight. A transdermal formulation comprising: the amounts of the polyol and the fatty acid are selected such that upon administration an effective amount of the NO booster is delivered transdermally. Transdermal formulations.

2. 2. The transdermal formulation of claim 1, wherein the curcuminoid is curcumin.

3. 10. The transdermal formulation of claim 1, wherein the formulation comprises curcumin and at least one of quercetin and apigenin.

4. 2. The transdermal formulation of claim 1, wherein the curcuminoid and the at least one flavonoid are in a ratio ranging from about 1:1 to about 10:

1.

5. 10. The transdermal formulation of claim 1, wherein the formulation comprises quercetin and apigenin in a ratio ranging from about 1:1 to about 10:

1.

6. 2. The transdermal formulation of claim 1, wherein the fatty acid is myristic acid.

7. 2. The transdermal formulation of claim 1, wherein the polyol is selected from the group consisting of polyethylene glycol, polypropylene glycol, ethylene glycol, propylene glycol, and glycerol.

8. 2. The transdermal formulation of claim 1, wherein the polyol is a polyethylene glycol having a molecular weight ranging from 200 to about 600.

9. 10. The transdermal formulation of claim 1, comprising a curcuminoid in the range of about 3% to about 10% by weight.

10. 2. The transdermal formulation of claim 1, wherein the amounts of the curcuminoid and the polyol are selected to provide a sustained release of the curcuminoid over a period of about 15 hours.

11. 11. The transdermal formulation of claim 10, wherein the polyol and the curcuminoid are in a ratio ranging from about 8:1 to about 12:

1.

12. 1. Use of a transdermal formulation for the manufacture of a medicament for treating a disease or condition associated with elevated CD38 levels, comprising: The transdermal formulation comprises: (a) an effective amount of a NO booster comprising one or both of an effective amount of a curcuminoid and an effective amount of at least one flavonoid, wherein the curcuminoid is selected from the group consisting of curcumin, demethoxycurcumin, and bisdemethoxycurcumin; (b) a polyol in an amount sufficient to dissolve the effective amount of curcuminoid and the at least one flavonoid; and optionally (c) a fatty acid, wherein the polyol and the fatty acid are in a ratio ranging from about 10:1 to about 50:1 by weight. Including, the amounts of the polyol and the fatty acid are selected such that upon administration an effective amount of the NO booster is delivered transdermally. use.

13. 13. The use of claim 12, wherein the disease or condition is selected from lupus, rheumatoid arthritis, multiple sclerosis, leukemia, multiple myeloma, neurodegenerative diseases, age-related physical and cognitive decline, chronic and acute inflammatory conditions, renal failure, coronary artery disease, heart failure, stroke, peripheral artery disease, myocardial infarction, diabetes, hypertension, neuropathic pain, osteoarthritis, anemia of chronic disease, ALS, Parkinkinson's, ischemia-reperfusion injury, hypoxia-reoxygenation injury, transfusion-induced injury, radiation-induced injury including dermatitis, and neuroinflammation.

14. 13. The use according to claim 12, wherein the disease or condition is selected from the group consisting of dementia, Alzheimer's disease (AD), and Parkinson's disease.

15. 13. The use of claim 12, wherein the disease or condition is selected from the group consisting of lupus, rheumatoid arthritis, multiple sclerosis, leukemia, and multiple myeloma.

16. 13. The use according to claim 12, wherein the disease or condition is type 2 diabetes.

17. 1. Use of a transdermal formulation for the manufacture of a medicament for reducing CD38 activity, comprising: The transdermal formulation comprises: (a) an effective amount of a NO booster comprising one or both of an effective amount of a curcuminoid and an effective amount of at least one flavonoid, wherein the curcuminoid is selected from the group consisting of curcumin, demethoxycurcumin, and bisdemethoxycurcumin; (b) a polyol in an amount sufficient to dissolve the effective amount of curcuminoid and the at least one flavonoid; and optionally (c) a fatty acid, wherein the polyol and the fatty acid are in a ratio ranging from about 10:1 to about 50:1 by weight. Including, the amounts of the polyol and the fatty acid are selected such that upon administration an effective amount of the NO booster is delivered transdermally. use.

18. The use of claim 17, wherein the CD38 is in a cell.

19. 20. The use of claim 18, wherein said contacting occurs in vitro.

20. 20. The use of claim 18, wherein the contacting occurs in vivo.

21. 13. The use of claim 12, wherein the formulation comprises curcumin and at least one of quercetin and apigenin.

22. 22. The use of claim 21, wherein the polyol is a polyethylene glycol having a molecular weight in the range of 200 to about 600.

23. 18. The use of claim 17, wherein the formulation comprises curcumin and at least one of quercetin and apigenin.

24. 24. The use of claim 23, wherein the polyol is a polyethylene glycol having a molecular weight in the range of 200 to about 600.

Images