Topical l-arginine compositions

Abstract

The present disclosure describes topical L-arginine compositions that comprise a deep eutectic solvent, such as L-arginine and oleic acid, and methods and uses thereof. The compositions of the present disclosure are capable of delivering an effective amount of L-arginine through the skin and are believed to be useful in vasodilation, such as to increase blood flow in extremities of diabetes patients.

Description

Attorney Docket No. 063594-504001WOTOPICAL L-ARGININE COMPOSITIONSCROSS REFERENCE TO RELATED APPLICATIONS

[0001] This application claims priority to U.S. Provisional Application No. 63 / 749,170, filed January 24, 2025; and U.S. Provisional Application No. 63 / 824,721, filed June 16, 2025, each of which is incorporated herein in its entirety for all purposes.BACKGROUND

[0002] L-Arginine is an amino acid naturally found in red meat, poultry, fish, and dairy. L-Arginine is converted in the body into nitric oxide which acts as a vasodilator. Nitric oxide causes blood vessels to open wider for improved blood flow. L-Arginine also stimulates the release of growth hormone, insulin, and other substances in the body. It is available in over-the-counter supplements. L-Arginine may cause side effects such as stomach pain, bloating, diarrhea, and low blood pressure.

[0003] Deep eutectic solvents, also known as eutectic solvents or DES, have a lower melting point than each of their individual components due to ion-dipole interactions and / or hydrogen bonding. DES are believed to be more environmentally friendly because often the components of the mixtures have been toxicologically evaluated. See, e.g., US patent publication numbers US 20210299044 and 20210308048.

[0004] Topical administration of drugs can have advantages, such as enhanced delivery of the active ingredient to the local desired site of action and reduced adverse side effects, compared to oral administration. See, e.g., Mehta, et al. Current Opinion Rheumatology 2021, 33(1): 94-109. For instance, drug delivery systems involving hydrogels, liposomes, and particle-based carriers have been investigated for providing effective relief of osteoarthritis symptoms. Such systems may suffer from high cost of goods in their development.

[0005] Drug penetration at therapeutically effective doses, or lack thereof, is a challenge for those in the pharmaceutical industry developing topically administered active ingredients. Many solubilizing agents have come under study, including DESs. Recently, research groups have investigated a variety of DES combinations and poorly-soluble drugs. This class of DESs hasAttorney Docket No. 063594-504001WObeen investigated with the goal of overcoming certain shortcomings, such as poor solubility, low absorption rate, and high toxicity, of drugs alone.

[0006] L-Arginine is a naturally occurring zwitterionic amino acid. Due to its charged nature, L-arginine typically cannot penetrate hydrophobic barriers such as the skin in an amount suitable for therapeutic use. Deep eutectic solvents have been used to deliver active ingredients across the skin. The physicochemical properties of a specific active ingredient such as L-arginine must be matched with those of the DES in order to deliver therapeutically effective doses for topical treatment, such as for local vasodilation to increase blood flow in the extremities of diabetes patients. This invention solves this problem among others.BRIEF SUMMARY

[0007] In some embodiments, the present invention provides a composition comprising: (a) from about 1% to about 50% (w / w) L-arginine; and (b) from about 3% to about 90% (w / w) oleic acid; wherein the L-arginine and the oleic acid form a mixture having a lower melting point than pure oleic acid. For instance, composition can comprise from about 3% to about 40% (w / w) L-arginine. In some embodiments, the composition comprises from about 5% to about 50% (w / w) oleic acid. The composition can further comprise additional ingredients. For example, the composition can comprise from about 2% to about 20% (w / w) caprylic acid. The composition can comprise from about 2% to about 20% (w / w) lauric acid. The composition can comprise from about 2% to about 20% (w / w) levulinic acid. The composition can comprise from about 5% to about 20% (w / w) glyceryl monostearate. The composition can comprise from about 10% to about 30% (w / w) isopropyl myristate. The composition can comprise from about 2% to about 20% (w / w) squalane. The composition can comprise less than about 5% (w / w) dimethyl sulfoxide. The composition can comprise from about 1% to about 2% (w / w) eucalyptol. The composition can be substantially free of water. In some embodiments, the composition comprises (a) from about 10% to about 15% (w / w) L-arginine; (b) from about 10% to about 30% (w / w) oleic acid; (c) from about 2% to about 10% (w / w) caprylic acid; (d) from about 2% to about 20% (w / w) lauric acid; (e) from about 8% to about 16% (w / w) levulinic acid; (f) from about 7% to about 15% (w / w) glyceryl monostearate; (g) from about 10% to about 30% (w / w) isopropyl myristate; (h) from about 2% to about 20% (w / w) squalane; (i) from about 0.5% to about 1%Attorney Docket No. 063594-504001WO(w / w) dimethyl sulfoxide; and (j) from about 1% to about 2% (w / w) eucalyptol; wherein the L-arginine and the oleic acid form a mixture having a lower melting point than pure oleic acid.

[0008] In some embodiments, the method of the present disclosure is a method of enhancing blood flow in a human subject in need thereof, comprising topically administering to the subject a composition as described herein.

[0009] In some embodiments, a method of the present disclosure can be a method of enhancing blood flow in an extremity in a human subject in need thereof, comprising topically administering to the extremity a composition as described herein. For instance, the method can comprise administering about 5 g of the composition. The method can comprise administering three times daily. The method can comprise administering about every four hours. The method can comprise administering for at least about eight days. In some embodiments, the extremity comprises a lower leg. In some embodiments, the enhanced blood flow is compared to an untreated extremity. The method can comprise increasing blood flow by from about 10% to about 1000% compared to blood flow in an untreated extremity

[0010] The specification provides solutions to these challenges and provides additional benefits as well.BRIEF DESCRIPTION OF THE DRAWINGS

[0011] FIG. 1 shows posterior tibialis mean blood flow from baseline difference on the first day: y-axis: blood flow from baseline (mL / min); x-axis = time. Mean T = mean treated; Mean U = mean untreated.

[0012] FIG. 2 shows posterior tibialis mean blood flow from baseline difference over duration of study: y-axis: blood flow from baseline (mL / min); x-axis = visit number (X.O = beginning of day; X.5 = end of day). Mean T = mean treated; Mean U = mean untreated.

[0013] FIG. 3 shows posterior tibialis mean change blood flow from baseline difference: y-axis: % blood flow change from baseline (visit 1); x-axis = visit number (X.O = beginning of day; X.5 = end of day). Mean T = mean treated; Mean U = mean untreated.Attorney Docket No. 063594-504001WO

[0014] FIG. 4 shows dorsalis pedis mean blood flow from baseline difference on the first day: y-axis: blood flow from baseline (mL / min); x-axis = time. Mean T = mean treated; Mean U = mean untreated.

[0015] FIG. 5 shows dorsalis pedis mean blood flow from baseline difference over duration of study: y-axis: blood flow from baseline (mL / min); x-axis = visit number (X.O = beginning of day; X.5 = end of day). Mean T = mean treated; Mean U = mean untreated.DETAILED DESCRIPTIONI. GENERAL

[0016] The present disclosure describes topical compositions of L-arginine that comprise a eutectic solvent, such as L-arginine and oleic acid. Such topical compositions are capable of delivering an effective amount of L-arginine to below the skin surface, e.g., to blood vessels at the site of administration, for vasodilation, e.g., in a diabetes patient. The topical compositions of the present disclosure are believed advantageous compared to other modes of delivering L-arginine known in the art, such as oral delivery of L-arginine, by enhancing local delivery, thereby reducing potential adverse side effects of taking high oral dosages of L-arginine.

[0017] Deep eutectic solvents have been investigated for use in transcutaneous delivery of polar drugs, such as non-steroidal anti-inflammatory drugs (NSAIDs). See, e.g., US patent publication numbers US 20210299044 and 20210308048. However, not all deep eutectic solvent combinations can deliver a therapeutically effective amount of every drug. In fact, the physicochemical properties of the deep eutectic solvent must be matched with those of the active drug for effective transcutaneous delivery of a sufficient amount of the drug for therapy.

[0018] L-Arginine was expected to be difficult to prepare as a therapeutically effective transcutaneous composition. As an amino acid, L-arginine is a polar molecule with ionizable groups that cannot easily traverse the lipid skin layers in sufficient quantities.

[0019] The present invention describes L-arginine compositions containing oleic acid. See, Example 1. In some embodiments, the composition is a viscous liquid at room temperature. Such compositions are capable of delivering therapeutically effective doses of the active ingredientAttorney Docket No. 063594-504001WOtranscutaneously in in vitro studies in contrast to commercially available topical compositions of L-arginine. See, Example 2.

[0020] In an illustrative embodiment, Composition F140-009IPA was administered 5 g per dose three times daily at 0, 4, and 8 hours over eight days to a lower leg in healthy elderly volunteers with reduced circulation in the extremities. See, Example 3. As measured by Doppler ultrasound on the posterior tibial and the dorsalis pedis arteries, blood flow showed a trend to improvement over a single day (see, FIG. 1 and 4) and over the course of the study (FIG. 2 and 5). Eight out of ten participants showed significant improvement based on change in blood flow as measured by Doppler ultrasound at the posterior tibialis artery. Mean difference between treated and untreated leg was 26% on Day 8, achieving statistical significance with p-value of 0.016. See, FIG. 3.

[0021] The data in the Examples suggest that the L-arginine compositions of the present disclosure are capable of delivering a therapeutically effective dose of the active ingredient for daily administration for vasodilation, such as for treating reduced blood flow in the extremities of diabetes patients.II. DEFINITIONS

[0022] Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art.

[0023] “Pharmaceutically effective amount” refers to an amount of the compound of the present disclosure in a composition or combination thereof, that provides the desired therapeutic or pharmaceutical result.

[0024] “Pharmaceutically acceptable excipient” includes without limitation any adjuvant, carrier, excipient, glidant, sweetening agent, diluent, preservative, dye / colorant, flavor enhancer, surfactant, wetting agent, dispersing agent, suspending agent, stabilizer, isotonic agent, solvent, or emulsifier which has been approved by the United States Food and Drug Administration as being acceptable for use in humans or domestic animals.

[0025] “Treatment” or “treat” or “treating” as used herein refers to an approach for obtaining beneficial or desired results. For purposes of the present disclosure, beneficial or desired resultsAttorney Docket No. 063594-504001WOinclude, but are not limited to, alleviation of a symptom and / or diminishment of the extent of a symptom and / or preventing a worsening of a symptom associated with a disease or condition. In one embodiment, “treatment” or “treating” includes one or more of the following: a) inhibiting the disease or condition (e.g., decreasing one or more symptoms resulting from the disease or condition, and / or diminishing the extent of the disease or condition); b) slowing or arresting the development of one or more symptoms associated with the disease or condition e.g., stabilizing the disease or condition, delaying the worsening or progression of the disease or condition); and c) relieving the disease or condition, e.g., causing the regression of clinical symptoms, ameliorating the disease state, delaying the progression of the disease, increasing the quality of life, and / or prolonging survival.

[0026] “Therapeutically effective amount” or “effective amount” as used herein refers to an amount that is effective to elicit the desired biological or medical response, including the amount of the compound that, when administered to a subject for treating a disease, is sufficient to effect such treatment for the disease. The effective amount will vary depending on the compound, the disease, and its severity and the age, weight, etc., of the subject to be treated. The effective amount can include a range of amounts. As is understood in the art, an effective amount may be in one or more doses, i.e., a single dose or multiple doses may be required to achieve the desired treatment endpoint. An effective amount may be considered in the context of administering one or more therapeutic agents, and a single agent may be considered to be given in an effective amount if, in conjunction with one or more other agents, a desirable or beneficial result may be or is achieved. Suitable doses of any co-administered compounds may optionally be lowered due to the combined action (e.g., additive or synergistic effects) of the compounds.

[0027] “Subject” is any mammal, such as a mouse, a rat, a dog, a cat, including veterinary animals, such as a goat, a pig, a horse, a cow, or a donkey, and primates, such as non-human primates, e.g., a cynomolgous monkey, rhesus monkey, or chimpanzee, as well as humans. In some embodiments, the subject is a human. In some embodiments, a subject is a patient.III. COMPOUNDS

[0028] The present compositions comprise L-arginine. L-Arginine, also known as L-Arg, Arg, or L-2-amino-5-guanidinopentanoic acid, is the amino acid with the formulaAttorney Docket No. 063594-504001WO(H2N)(HN)CN(H)(CH2)3CH(NH2)CO2H. The molecule features a guanidino group appended to a standard amino acid framework. At physiological pH, the carboxylic acid is deprotonated (-CO2 ) and both the amino and guanidino groups are protonated, resulting in a cation. Pure L-arginine has a melting point of about 260 °C.

[0029] L-Arginine has a chemical structure:

[0030] In some embodiments, the composition comprises oleic acid. Oleic acid, also known as (9Z)-octadec-9-enoic acid, (Z)-octadec-9-enoic acid, cA-9-octadecenoic acid, or cis-9-octadecenoic acid, is a fatty acid that occurs naturally in various animal and vegetable fats and oils. It is an odorless, colorless oil, although commercial samples may be yellowish due to the presence of impurities. Oleic acid is classified as a monounsaturated omega-9 fatty acid, abbreviated with a lipid number of 18: 1 cis-9, and a main product of A9-desaturase. It has the formula CH3-(CH2)7_CH=CH-(CH2)7_COOH. Pure oleic acid has a melting point of from about 13 °C to about 16 °C.

[0031] Oleic acid has a chemical structure:

[0032] Caprylic acid (octanoic acid) is a medium-chain fatty acid that is naturally found in palm oil, coconut oil, and the milk of humans and some animals.

[0033] Caprylic acid has a chemical structure:

[0034] Lauric acid, also known as dodecanoic acid, is a saturated fatty acid with a 12-carbon atom chain.Attorney Docket No. 063594-504001WO

[0035] Lauric acid has a chemical structure:

[0036] Levulinic acid, or 4-oxopentanoic acid, is an organic compound withthe formula CH3C(O)CH2CH2CO2H. Levulinic acid is a white crystalline solid is soluble in water and polar organic solvents. It can be derived from degradation of cellulose.

[0037] Levulinic acid has a chemical structure:

[0038] Stearic acid, or octadecanoic acid, is a saturated fatty acid with an 18-carbon chain having the formula CH3(CH2)ieCO2H. As a pure compound at room temperature, it is a soft waxy solid with a melting point of about 69 °C and pKa of 4.5.

[0039] Stearic acid has a chemical structure:

[0040] Isostearic acid, also known as iso-octadecanoic acid or 16-methylheptadecanoic acid, is an branched saturated fatty acid that can be derived from vegetable oils. Isostearic acid has the molecular formula (CFLXCEb sCChH.

[0041] Iso stearic acid has a chemical structure:IV. COMPOSITIONS

[0042] The present disclosure describes topical compositions of L-arginine that comprise a eutectic solvent. Such compositions demonstrate enhanced delivery of L-arginine across membranes, such as the skin. L-arginine may serve as the pharmaceutically active ingredient asAttorney Docket No. 063594-504001WOwell as one member of a eutectic solvent pair, e.g, in the combination of L-arginine and oleic acid.

[0043] In some embodiments, a composition of the present disclosure comprises:(a) from about 1% to about 50% (w / w) L-arginine; and(b) from about 3% to about 90% (w / w) oleic acid;wherein the L-arginine and the oleic acid form a mixture having a lower melting point than pure oleic acid.

[0044] In some embodiments, a composition of the present disclosure comprises:(a) from about 3% to about 40% (w / w) L-arginine; and(b) from about 3% to about 90% (w / w) oleic acid;wherein the L-arginine and the oleic acid form a mixture having a lower melting point than pure oleic acid.

[0045] The composition of the present disclosure comprises a deep eutectic mixture of L-arginine and oleic acid that serves as a base to incorporate the other ingredients of the topical composition. In some embodiments, the deep eutectic mixture is prepared by mixing the L-arginine and oleic acid at elevated temperature, e.g., from about 50 °C to about 110 °C, such as from about 60 °C to about 90 °C or from about 50 °C to about 80 °C, before combining with the remaining ingredients in the composition. Accordingly, in some embodiments, the L-arginine and the oleic acid in the present composition form a mixture having a lower melting point of from about 0.01 °C to about 10 °C, such as from about 0.01 °C to about 8 °C, about 0.01 °C to about 6 °C, about 0.01 °C to about 5 °C, about 0.01 °C to about 4 °C, about 0.01 °C to about 3 °C, about 0.01 °C to about 2 °C, about 0.01 °C to about 1 °C, as compared to pure oleic acid. In some embodiments, the L-arginine and the oleic acid in the present composition form a mixture having a lower melting point of about 0.01 °C, 0.02 °C, 0.03 °C, 0.04 °C, 0.05 °C, 0.06 °C, 0.07 °C, 0.08 °C, 0.09 °C, 0.1 °C, 0.2 °C, 0.3 °C, 0.4 °C, 0.5 °C, 0.6 °C, 0.7 °C, 0.8 °C, 0.9 °C, 1 °C, 2 °C, 3 °C, 4 °C, 5 °C, 6 °C, 7 °C, 8 °C, 9 °C, or about 10 °C as compared to pure oleic acid. Preparations of exemplary compositions of the present disclosure are described in Example 1 herein.Attorney Docket No. 063594-504001WO

[0046] As understood in the art, the term “about” means approximately. Tn some embodiments, the term “about” when referring to a value includes the stated value + / - 10% of the stated value. For example, about 50% can include a range of from 45% to 55%, while about 20 molar equivalents can include a range of from 18 to 22 molar equivalents. Accordingly, in some embodiments, when referring to a range, “about” refers to each of the stated values + / - 10% of the stated value of each end of the range. For instance, a ratio of from about 1 to about 3 (weight / weight) can include a range of from 0.9 to 3.3.

[0047] In some embodiments, the composition comprises from about 3% to about 40% (w / w), such as from about 3% to about 30%, from about 3% to about 25%, from about 3% to about 20%, from about 3% to about 15%, from about 5% to about 15%, or from about 10% to about 15% (w / w) L-arginine. In some embodiments, the composition comprises about 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, or about 20% (w / w) L-arginine. In some embodiments, the composition comprises about 3% to about 40% (w / w) L-arginine. In some embodiments, the composition comprises about 14% (w / w) L-arginine.

[0048] In some embodiments, the composition comprises from about 5% to about 90% (w / w), such as from about 5% to about 80%, from about 5% to about 70%, from about 5% to about 60%, from about 5% to about 50%, from about 5% to about 40%, from about 5% to about 30%, from about 5% to about 25%, from about 10% to about 30%, from about 15% to about 25%, or from about 15% to about 20% (w / w) oleic acid. In some embodiments, the composition comprises about 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, or about 30% (w / w) oleic acid. In some embodiments, the composition comprises from about 5% to about 50% (w / w) oleic acid. In some embodiments, the composition comprises about 15% (w / w) oleic acid. In some embodiments, the composition comprises about 19% (w / w) oleic acid. In some embodiments, the composition comprises about 20% (w / w) oleic acid.

[0049] In some embodiments, a composition of the present disclosure comprises:(a) from about 3% to about 30% (w / w) L-arginine; and(b) from about 10% to about 30% (w / w) oleic acid;wherein the L-arginine and the oleic acid form a mixture having a lower melting point than pure oleic acid.Attorney Docket No. 063594-504001WO

[0050] In some embodiments, a composition of the present disclosure comprises:(a) from about 10% to about 15% (w / w) L-arginine; and(b) from about 10% to about 30% (w / w) oleic acid;wherein the L-arginine and the oleic acid form a mixture having a lower melting point than pure oleic acid.

[0051] The composition can be one of many topical composition types containing an oil as the major ingredient, including solutions, gels, creams, sprays and foams. The composition can further include additional pharmaceutically acceptable excipients typically used in topical compositions and known to those skilled in the art. Such excipients include, for example, humectants, emollients, pH stabilizing agents, preservatives, chelating agents, and anti-oxidants. Illustrative excipients can be found n Remington: The Science and Practice of Pharmacy.Adejare, A., ed. Academic Press, 23th Edition (2020).

[0052] In some embodiments, the composition of the present disclosure is an emulsion, e.g., to form a cream, lotion, or other suitable composition. An emulsion typically includes a first phase (e.g., a discontinuous phase) contained within a second fluid phase (e.g., a continuous phase). In some embodiments, the emulsion may take the form of a cream or a lotion.

[0053] As an example, a cream may include materials such as oils, triglycerides, stearates, fatty acids, fatty alcohols, squalenes, polysorbates, or the like. In some embodiments, such materials are hydrophobic, which can be emulsified with water or other aqueous phases, e.g., to produce an emulsion. In one embodiment, for example, the cream may include a saturated squalene. Examples of stearates include, but are not limited to, glyceryl stearate, propylene glycol stearate, steryl stearate, sorbitan stearate, sodium stearate, calcium stearate, magnesium steratae, glycol sterate, and the like. Non-limiting examples of oils include mineral oil, wheat germ oil, palm oil, nut oil, linseed oil, coconut oil, etc. Other materials may also be present within the composition, for example, buffers, preservatives, surfactants, etc.

[0054] In some embodiments, as a non-limiting example, a cream can include one or more of water, mineral oil, glyceryl stereate, squalene, propylene glycol stearate, wheat germ oil, glyceryl stearate, isopropyl myristate, steryl stearate, polysorbate 60, propylene glycol, oleic acid, tocopherol acetate, collagen, sorbitan stearate, vitamin A and D, triethanolamine, methylparaben,Attorney Docket No. 063594-504001WOaloe vera extract, imidazolidinyl urea, propylparaben, PND, and / or BHA. Tn some embodiments, a cream can include one or more of mineral oil, caprylic / capric triglycerides, phenoxyethanol, glycerol stearate, PEG 75 stearate, cetyl alcohol, methylparaben, and propylparaben.

[0055] In some embodiments, the composition comprises from about 2% to about 20% (w / w), such as from about 2% to about 10%, from about 10% to about 20%, from about 3% to about 15%, from about 10% to about 15%, or from about 5% to about 10% (w / w) caprylic acid. In some embodiments, the composition comprises from about 2% to about 20% (w / w) caprylic acid. In some embodiments, the composition comprises about 2%, 3%, 4%, 5%, 6%, 6.1%, 6.2%, 6.3%, 6.4%, 6.5%, 6.6%, 6.7%, 6.8%, 6.9%, 7%, 7.1%, 7.2%, 7.3%, 7.4%, 7.5%, 7.6%, 7.7%, 7.8%, 7.9%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, or about 20% (w / w) caprylic acid. In some embodiments, the composition comprises about 6.1% (w / w) caprylic acid.

[0056] In some embodiments, a composition of the present disclosure comprises:(a) from about 3% to about 30% (w / w) L-arginine;(b) from about 10% to about 30% (w / w) oleic acid; and(c) from about 2% to about 10% (w / w) caprylic acid;wherein the L-arginine and the oleic acid form a mixture having a lower melting point than pure oleic acid.

[0057] In some embodiments, a composition of the present disclosure comprises:(a) from about 10% to about 15% (w / w) L-arginine;(b) from about 10% to about 30% (w / w) oleic acid; and(c) from about 2% to about 10% (w / w) caprylic acid;wherein the L-arginine and the oleic acid form a mixture having a lower melting point than pure oleic acid.

[0058] In some embodiments, the composition comprises from about 2% to about 20% (w / w), such as from about 2% to about 10%, from about 10% to about 20%, from about 3% to about 15%, from about 10% to about 15%, or from about 5% to about 10% (w / w) lauric acid. In some embodiments, the composition comprises about 2%, 3%, 4%, 5%, 6%, 7%, 7.1%, 7.2%, 7.3%, 7.4%, 7.5%, 7.6%, 7.7%, 7.8%, 7.9%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%,Attorney Docket No. 063594-504001WO18%, 19%, or about 20% (w / w) lauric acid. In some embodiments, the composition comprises from about 2% to about 20% (w / w) lauric acid. In some embodiments, the composition comprises about 7.5% (w / w) lauric acid.

[0059] In some embodiments, the composition comprises from about 2% to about 20% (w / w), such as from about 2% to about 10%, from about 10% to about 20%, from about 3% to about 15%, from about 10% to about 15%, or from about 5% to about 10% (w / w) stearic acid. In some embodiments, the composition comprises about 2%, 3%, 4%, 5%, 6%, 7%, 7.1%, 7.2%, 7.3%, 7.4%, 7.5%, 7.6%, 7.7%, 7.8%, 7.9%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, or about 20% (w / w) stearic acid. In some embodiments, the composition comprises from about 2% to about 20% (w / w) stearic acid. In some embodiments, the composition comprises about 7.5% (w / w) stearic acid.

[0060] In some embodiments, a composition of the present disclosure comprises:(a) from about 3% to about 30% (w / w) L-arginine;(b) from about 10% to about 30% (w / w) oleic acid;(c) from about 2% to about 10% (w / w) caprylic acid; and(d) from about 2% to about 20% (w / w) lauric acid;wherein the L-arginine and the oleic acid form a mixture having a lower melting point than pure oleic acid.

[0061] In some embodiments, a composition of the present disclosure comprises:(a) from about 10% to about 15% (w / w) L-arginine;(b) from about 10% to about 30% (w / w) oleic acid;(c) from about 2% to about 10% (w / w) caprylic acid; and(d) from about 2% to about 20% (w / w) lauric acid;wherein the L-arginine and the oleic acid form a mixture having a lower melting point than pure oleic acid.

[0062] In some embodiments, the composition comprises from about 2% to about 20% (w / w), such as from about 2% to about 10%, from about 10% to about 20%, from about 8% to about 16%, from about 3% to about 15%, from about 10% to about 15%, or from about 2% to about 5% (w / w) levulinic acid. In some embodiments, the composition comprises about 2%, 3%, 4%,Attorney Docket No. 063594-504001WO5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, or about 20% (w / w) levulinic acid. In some embodiments, the composition comprises from about 2% to about 20% (w / w) levulinic acid. In some embodiments, the composition comprises about 10% (w / w) levulinic acid. In some embodiments, the composition comprises about 14% (w / w) levulinic acid.

[0063] In some embodiments, the composition comprises from about 2% to about 20% (w / w), such as from about 2% to about 10%, from about 10% to about 20%, from about 8% to about 16%, from about 3% to about 15%, from about 10% to about 15%, or from about 2% to about 5% (w / w) isostearic acid. In some embodiments, the composition comprises about 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, or about 20% (w / w) isostearic acid. In some embodiments, the composition comprises from about 2% to about 20% (w / w) isostearic acid. In some embodiments, the composition comprises about 10% (w / w) isostearic acid. In some embodiments, the composition comprises about 14% (w / w) isostearic acid.

[0064] In some embodiments, a composition of the present disclosure comprises:(a) from about 3% to about 30% (w / w) L-arginine;(b) from about 10% to about 30% (w / w) oleic acid;(c) from about 2% to about 10% (w / w) caprylic acid;(d) from about 2% to about 20% (w / w) lauric acid; and(e) from about 8% to about 16% (w / w) levulinic acid;wherein the L-arginine and the oleic acid form a mixture having a lower melting point than pure oleic acid.

[0065] In some embodiments, a composition of the present disclosure comprises:(a) from about 10% to about 15% (w / w) L-arginine;(b) from about 10% to about 30% (w / w) oleic acid;(c) from about 2% to about 10% (w / w) caprylic acid;(d) from about 2% to about 20% (w / w) lauric acid; and(e) from about 8% to about 16% (w / w) levulinic acid;Attorney Docket No. 063594-504001WOwherein the L-arginine and the oleic acid form a mixture having a lower melting point than pure oleic acid.

[0066] Compositions provided herein may comprise inorganic or organic buffers (e.g., sodium or potassium salts of phosphate, carbonate, acetate, or citrate) and pH adjustment agents (e g., hydrochloric acid, sodium or potassium hydroxide, salts of citrate or acetate, amino acids and their salts) antioxidants (e.g., ascorbic acid, alpha- tocopherol), surfactants (e.g., polysorbate 20, polysorbate 80, polyoxyethylene9-10 nonyl phenol, sodium desoxy cholate), solution and / or cryo / lyo stabilizers (e.g., sucrose, lactose, mannitol, trehalose), osmotic adjustment agents (e.g., salts or sugars), antibacterial agents (e.g., benzoic acid, phenol, gentamicin), antifoaming agents (e.g., polydimethylsilozone), preservatives (e.g., thimerosal, 2-phenoxyethanol, EDTA), polymeric stabilizers and viscosity-adjustment agents (e.g., polyvinylpyrrolidone, poloxamer 488, carboxymethylcellulose) and co-solvents (e.g., glycerol, polyethylene glycol, ethanol).

[0067] In some embodiments, the composition further comprises an emulsifier. Emulsifiers are added to creams and lotions to form homogenous mixtures of both. Addition of an emulsifier not only stabilizes the emulsion but also influences the consistency of the composition. Emulsifiers are used in creams, lotions, sprays and foams and are categorized as oil in water or water in oil. Oil in water emulsifiers keep oil drops packed in water and are used more in moisturizing products; water in oil emulsifiers keep water droplets packed in oil and are used for a fatty feel. Emulsifiers include lecithin, glyceryl stearate and cetearyl alcohol.

[0068] In some embodiments, the composition further comprises squalane. In some embodiments, the composition comprises from about 5% to about 30%, such as from about 5% to about 20%, from about 7% to about 15%, or from about 10% to about 20% (w / w) glyceryl monostearate. In some embodiments, the composition comprises from about 5% to about 20% (w / w) glyceryl monostearate. In some embodiments, the composition comprises about 5%. 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, or about 20% (w / w) glyceryl monostearate. In some embodiments, the composition comprises about 14% (w / w) glyceryl monostearate.

[0069] In some embodiments, a composition of the present disclosure comprises:(a) from about 3% to about 30% (w / w) L-arginine;Attorney Docket No. 063594-504001WO(b) from about 10% to about 30% (w / w) oleic acid;(c) from about 2% to about 10% (w / w) caprylic acid;(d) from about 2% to about 20% (w / w) lauric acid;(e) from about 8% to about 16% (w / w) levulinic acid; and(f) from about 7% to about 15% (w / w) glyceryl monostearate;wherein the L-arginine and the oleic acid form a mixture having a lower melting point than pure oleic acid.

[0070] In some embodiments, a composition of the present disclosure comprises:(a) from about 10% to about 15% (w / w) L-arginine;(b) from about 10% to about 30% (w / w) oleic acid;(c) from about 2% to about 10% (w / w) caprylic acid;(d) from about 2% to about 20% (w / w) lauric acid;(e) from about 8% to about 16% (w / w) levulinic acid; and(f) from about 7% to about 15% (w / w) glyceryl monostearate;wherein the L-arginine and the oleic acid form a mixture having a lower melting point than pure oleic acid.

[0071] In some embodiments, the composition comprises from about 10% to about 60% (w / w), such as from about 10% to about 55%, from about 10% to about 50%, from about 10% to about 40%, from about 10% to about 30%, or from about 15% to about 25% (w / w) isopropyl myristate. In some embodiments, the composition comprises from about 10% to about 30% (w / w) isopropyl myristate. In some embodiments, the composition comprises about 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, or about 30% (w / w) isopropyl myristate. In some embodiments, the composition comprises about 20% (w / w) isopropyl myristate.

[0072] In some embodiments, a composition of the present disclosure comprises:(a) from about 3% to about 30% (w / w) L-arginine;(b) from about 10% to about 30% (w / w) oleic acid;(c) from about 2% to about 10% (w / w) caprylic acid;(d) from about 2% to about 20% (w / w) lauric acid;Attorney Docket No. 063594-504001WO(e) from about 8% to about 16% (w / w) levulinic acid;(f) from about 7% to about 15% (w / w) glyceryl monostearate; and(g) from about 10% to about 30% (w / w) isopropyl myristate;wherein the L-arginine and the oleic acid form a mixture having a lower melting point than pure oleic acid.

[0073] In some embodiments, a composition of the present disclosure comprises:(a) from about 10% to about 15% (w / w) L-arginine;(b) from about 10% to about 30% (w / w) oleic acid;(c) from about 2% to about 10% (w / w) caprylic acid;(d) from about 2% to about 20% (w / w) lauric acid;(e) from about 8% to about 16% (w / w) levulinic acid;(f) from about 7% to about 15% (w / w) glyceryl monostearate; and(g) from about 10% to about 30% (w / w) isopropyl myristate;wherein the L-arginine and the oleic acid form a mixture having a lower melting point than pure oleic acid.

[0074] In some embodiments, the composition further comprises squalane. In some embodiments, the composition comprises from about 0.5% to about 30% (w / w), such as from about 5% to about 20% or from about 10% to about 20% (w / w), squalane. In some embodiments, the composition comprises from about 2% to about 20% (w / w) squalane. In some embodiments, the composition comprises about 0.5%. 0.6%, 0.7%, 0.8%, 0.9%, 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, or about 10% (w / w) squalane. In some embodiments, the composition comprises about 7% (w / w) squalane.

[0075] In some embodiments, a composition of the present disclosure comprises:(a) from about 3% to about 30% (w / w) L-arginine;(b) from about 10% to about 30% (w / w) oleic acid;(c) from about 2% to about 10% (w / w) caprylic acid;(d) from about 2% to about 20% (w / w) lauric acid;(e) from about 8% to about 16% (w / w) levulinic acid;(I) from about 7% to about 15% (w / w) glyceryl monostearate;Attorney Docket No. 063594-504001WO(g) from about 10% to about 30% (w / w) isopropyl myristate; and(h) from about 2% to about 20% (w / w) squalane;wherein the L-arginine and the oleic acid form a mixture having a lower melting point than pure oleic acid.

[0076] In some embodiments, a composition of the present disclosure comprises:(a) from about 10% to about 15% (w / w) L-arginine;(b) from about 10% to about 30% (w / w) oleic acid;(c) from about 2% to about 10% (w / w) caprylic acid;(d) from about 2% to about 20% (w / w) lauric acid;(e) from about 8% to about 16% (w / w) levulinic acid;(f) from about 7% to about 15% (w / w) glyceryl monostearate;(g) from about 10% to about 30% (w / w) isopropyl myristate; and(h) from about 2% to about 20% (w / w) squalane;wherein the L-arginine and the oleic acid form a mixture having a lower melting point than pure oleic acid.

[0077] In some embodiments, the emulsifier comprises a surfactant. In some embodiments, the emulsifier comprises polyethylene glycol monododecyl ether.

[0078] Cocinic acid, also known as coconut acid or 7 -hydroxy-9-m ethoxy-3 -methyl -2,4,4a,7,7a,13-hexahydro-lH-4,12-methanobenzofuro[3,2-e]isoquinoline-ll-carboxylic acid, is a fatty acid derived from coconut oil that can act as an emulsifier and / or surfactant. In some embodiments, the composition comprises cocinic acid. In some embodiments, the composition comprises from about 0.5% to about 30% (w / w), such as from about 5% to about 20% or from about 10% to about 20% (w / w), cocinic acid. In some embodiments, the composition comprises from about 2% to about 20% (w / w) cocinic acid. In some embodiments, the composition comprises from about 5% to about 10% (w / w) cocinic acid. In some embodiments, the composition comprises about 0.5%. 0.6%, 0.7%, 0.8%, 0.9%, 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, or about 10% (w / w) cocinic acid. In some embodiments, the composition comprises about 7% (w / w) cocinic acid.Attorney Docket No. 063594-504001WO

[0079] Span® 80, also known as sorbitan monooleate, is a sorbitan ester series of surfactant which is produced by esterification of one or more sorbitan hydroxyl group with fatty acid.

[0080] Polysorbate 80 (TWEEN 80) is a common fragrance and essential oil solubilizer as well as an emulsifier. It is soluble in water and alcohol and insoluble in oils. It is typically a coemulsifier that can be used both as a viscosity modifier and a dispersing agent.

[0081] In some embodiments, the composition comprises less than about 20% (w / w), such as less than about 15%, less than about 10%, less than about 5%, less than about 4%, less than about 3%, less than about 2%, or less than about 1%, of an alcohol, for example, glycerol, propylene glycol, or ethanol. In some embodiments, the composition is substantially free of an alcohol.

[0082] In some embodiments, the composition further comprises a viscosity-adjustment agent. Viscosity-adjustment agents, e.g., texture enhancers, can enhance the consistency, appearance, and feel of the composition. An illustrative viscosity-adjustment agent includes poly(methyl vinyl ether-alt-maleic acid monobutyl ester) (CAS #25119-68-0). In some embodiments, the composition further comprises poly(methyl vinyl ether-alt-maleic acid monobutyl ester).

[0083] In some embodiments, the composition comprises less than about 10% (w / w), such as less than about 5%, less than about 4%, less than about 3%, less than about 2%, or less than about 1%, of dimethyl sulfoxide. In some embodiments, the composition comprises less than about 5% (w / w) dimethyl sulfoxide.

[0084] In some embodiments, the composition further comprises dimethyl sulfoxide. In some embodiments, the composition comprises from about 0.1% to about 3% (w / w), such as from about 0.5% to about 2% or from about 0.1% to about 1.5% (w / w), dimethyl sulfoxide. In some embodiments, the composition comprises about 0.1%, 0.2%, 0.3%, 0.4%, 0.5%. 0.6%, 0.7%, 0.8%, 0.9%, 1%, 2%, or about 3% (w / w) dimethyl sulfoxide. In some embodiments, the composition comprises about 1% (w / w) dimethyl sulfoxide. In some embodiments, the composition comprises from 0% to about 1% (w / w) dimethyl sulfoxide. In some embodiments, the composition is substantially free of dimethyl sulfoxide.

[0085] In some embodiments, a composition of the present disclosure comprises:Attorney Docket No. 063594-504001WO(a) from about 3% to about 30% (w / w) L-arginine;(b) from about 10% to about 30% (w / w) oleic acid;(c) from about 2% to about 10% (w / w) caprylic acid;(d) from about 2% to about 20% (w / w) lauric acid;(e) from about 8% to about 16% (w / w) levulinic acid;(f) from about 7% to about 15% (w / w) glyceryl monostearate;(g) from about 10% to about 30% (w / w) isopropyl myristate;(h) from about 2% to about 20% (w / w) squalane; and(i) from about 0.5% to about 1% (w / w) dimethyl sulfoxide;wherein the L-arginine and the oleic acid form a mixture having a lower melting point than pure oleic acid.

[0086] In some embodiments, a composition of the present disclosure comprises:(a) from about 10% to about 15% (w / w) L-arginine;(b) from about 10% to about 30% (w / w) oleic acid;(c) from about 2% to about 10% (w / w) caprylic acid;(d) from about 2% to about 20% (w / w) lauric acid;(e) from about 8% to about 16% (w / w) levulinic acid;(f) from about 7% to about 15% (w / w) glyceryl monostearate;(g) from about 10% to about 30% (w / w) isopropyl myristate;(h) from about 2% to about 20% (w / w) squalane; and(i) from about 0.5% to about 1% (w / w) dimethyl sulfoxide;wherein the L-arginine and the oleic acid form a mixture having a lower melting point than pure oleic acid.

[0087] In some embodiments, the composition further comprises eucalyptol. In some embodiments, the composition comprises from about 0.1% to about 3% (w / w), such as from about 0.5% to about 2%, from about 1% to about 2%, or from about 0.1% to about 1.5% (w / w), eucalyptol. In some embodiments, the composition comprises from about 1% to about 2% eucalyptol. In some embodiments, the composition comprises about 0.1%, 0.2%, 0.3%, 0.4%, 0.5%. 0.6%, 0.7%, 0.8%, 0.9%, 1%, 1.1%, 1.2%, 1.3%, 1.4%, 1.5%. 1.6%, 1.7%, 1.8%, 1.9%,Attorney Docket No. 063594-504001WO2%, or about 3% (w / w) eucalyptol. In some embodiments, the composition comprises about 1.4% (w / w) eucalyptol.

[0088] In some embodiments, a composition of the present disclosure comprises:(a) from about 3% to about 30% (w / w) L-arginine;(b) from about 10% to about 30% (w / w) oleic acid;(c) from about 2% to about 10% (w / w) caprylic acid;(d) from about 2% to about 20% (w / w) lauric acid;(e) from about 8% to about 16% (w / w) levulinic acid;(!) from about 7% to about 15% (w / w) glyceryl monostearate;(g) from about 10% to about 30% (w / w) isopropyl myristate;(h) from about 2% to about 20% (w / w) squalane;(i) from about 0.5% to about 1% (w / w) dimethyl sulfoxide; and(j) from about 1% to about 2% (w / w) eucalyptol;wherein the L-arginine and the oleic acid form a mixture having a lower melting point than pure oleic acid.

[0089] In some embodiments, a composition of the present disclosure comprises:(a) from about 10% to about 15% (w / w) L-arginine;(b) from about 10% to about 30% (w / w) oleic acid;(c) from about 2% to about 10% (w / w) caprylic acid;(d) from about 2% to about 20% (w / w) lauric acid;(e) from about 8% to about 16% (w / w) levulinic acid;(f) from about 7% to about 15% (w / w) glyceryl monostearate;(g) from about 10% to about 30% (w / w) isopropyl myristate;(h) from about 2% to about 20% (w / w) squalane;(i) from about 0.5% to about 1% (w / w) dimethyl sulfoxide; and(j) from about 1% to about 2% (w / w) eucalyptol;wherein the L-arginine and the oleic acid form a mixture having a lower melting point than pure oleic acid.

[0090] In some embodiments, a composition of the present disclosure comprises:Attorney Docket No. 063594-504001WO(a) from about 10% to about 15% (w / w) L-arginine;(b) from about 10% to about 30% (w / w) oleic acid;(c) from about 2% to about 10% (w / w) caprylic acid;(d) from about 5% to about 10% (w / w) lauric acid;(e) from about 2% to about 16% (w / w) levulinic acid;(f) from about 10% to about 15% (w / w) glyceryl monostearate;(g) from about 10% to about 20% (w / w) isopropyl myristate;(h) from about 2% to about 10% (w / w) squalane;(i) from 0% to about 1% (w / w) dimethyl sulfoxide; and(j) from about 0.5% to about 2% (w / w) eucalyptol;wherein the L-arginine and the oleic acid form a mixture having a lower melting point than pure oleic acid.

[0091] In some embodiments, a composition of the present disclosure comprises:(a) from about 10% to about 15% (w / w) L-arginine;(b) from about 10% to about 30% (w / w) oleic acid;(c) from about 5% to about 10% (w / w) cocinic acid;(d) from about 5% to about 25% (w / w) coconut oil;(e) from about 10% to about 15% (w / w) glyceryl monostearate;(f) from about 10% to about 20% (w / w) isopropyl myristate;(g) from about 2% to about 10% (w / w) squalane; and(h) from 0% to about 1% (w / w) dimethyl sulfoxide;wherein the L-arginine and the oleic acid form a mixture having a lower melting point than pure oleic acid.

[0092] In some embodiments, a composition of the present disclosure comprises:(a) about 14% (w / w) L-arginine;(b) about 19% (w / w) oleic acid;(c) about 6.1% (w / w) caprylic acid;(d) about 7.5% (w / w) lauric acid;(e) about 10% (w / w) levulinic acid;(f) about 14% (w / w) glyceryl monostearate;Attorney Docket No. 063594-504001WO(g) about 20% (w / w) isopropyl myristate;(h) about 7% (w / w) squalane;(i) about 1% (w / w) dimethyl sulfoxide; and(j) about 1.4% (w / w) eucalyptol;wherein the L-arginine and the oleic acid form a mixture having a lower melting point than pure oleic acid.

[0093] In some embodiments, the composition is substantially free of water.

[0094] In some embodiments, the composition is substantially free of dimethyl sulfoxide.

[0095] Exemplary compositions of the present disclosure include those in the tables below. Amounts shown are percent by weight (w / w).aalso contained 2% Arlacel 170.Attorney Docket No. 063594-504001WO“also contained 10% myristic acid;balso contained 4.84% menthol.Attorney Docket No. 063594-504001WO“F140004 also contained 4% capric acid, 1% propanoic acid;bF120005 also contained 2% capric acid, 4% myristic acid, and 5% stearic acidAttorney Docket No. 063594-504001WOc140-001-lHMK also contained 2% caffeine, 4% capric acid, 2.7% propionic acid, 1.5% rosemary oil

[0096] Composition F140-009IPA was substantially the same as described for Composition F140-009EtOH above, except that isopropanol was used in its preparation instead of ethanol.

[0097] In some embodiments, the composition of the present disclosure is a liquid at room temperature. In some embodiments, room temperature is from about 15 °C to about 30 °C, such as from about 20 °C to about 30 °C or from about 15 °C to about 25 °C.

[0098] The compositions that comprise L-arginine can be administered by any useful route and means, such as by topical administration, in various forms including but not limited to a liquid, patch, cream, lotion, or gel. Therapeutically effective amounts of the compound may include from about 0.00001 mg / kg body weight per day to about 10 mg / kg body weight per day, such as from about 0.0001 mg / kg body weight per day to about 10 mg / kg body weight per day, or such as from about 0.001 mg / kg body weight per day to about 1 mg / kg body weight per day, or such as from about 0.01 mg / kg body weight per day to about 1 mg / kg body weight per day, or such as from about 0.05 mg / kg body weight per day to about 0.5 mg / kg body weight per day, or such asAttorney Docket No. 063594-504001WOfrom about 0.3 mg to about 30 mg per day, or such as from about 30 mg to about 300 mg per day.

[0099] In some embodiments, the composition of the present disclosure is administered at about 5 g.

[0100] The compositions of the present disclosure can also be delivered as microspheres for slow release in the body. For example, microspheres can be formulated for administration via intradermal injection of drug-containing microspheres, which slowly release subcutaneously (see Rao, J. Biomater Sci. Polym. Ed. 7:623-645, 1995; as biodegradable and injectable gel compositions (see, e.g., Gao Pharm. Res. 12:857-863, 1995); or, as microspheres for oral administration (see, e.g., Eyles, J. Pharm. Pharmacol. 49:669-674, 1997). Both transdermal and intradermal routes afford constant delivery for weeks or months.

[0101] Lipid-based drug delivery systems include lipid solutions, lipid emulsions, lipid dispersions, self-emulsifying drug delivery systems (SEDDS) and self-microemulsifying drug delivery systems (SMEDDS). In particular, SEDDS and SMEDDS are isotropic mixtures of lipids, surfactants and co- surfactants that can disperse spontaneously in aqueous media and form fine emulsions (SEDDS) or microemulsions (SMEDDS). Lipids useful in the compositions of the present disclosure include any natural or synthetic lipids including, but not limited to, sesame seed oil, olive oil, castor oil, peanut oil, fatty acid esters, glycerol esters, Labrafil®, Labrasol®, Cremophor®, Solutol®, Tween®, Capryol®, Capmul®, Captex®, and Peceol®.V. METHODSA. Measurement Methods

[0102] The present L-arginine compositions are capable of delivering effective amounts of L-arginine across membranes, e.g., the skin. In some embodiments, the L-arginine compositions of the present disclosure exhibit measurable amounts of L-arginine across the skin.

[0103] Methods and techniques for measuring transdermal delivery of compounds are known in the art, including direct and indirect measurements as well as in vitro and in vivo methods. Direct in vitro methods include diffusion cell models and skin flap methods, e.g., isolated perfused porcine skin flap. Indirect in vitro methods include techniques such as measurement ofAttorney Docket No. 063594-504001WOthe rate of trans-epidermal water loss, which can be used to assess barrier integrity of the skin. Direct in vivo methods include measurement of pharmacological response.

[0104] In vitro methods of directly measuring transdermal delivery include diffusion cell models. A wide variety of diffusion cell systems have been developed for measuring compound permeation through membranes, including vertical cells, where the donor chamber is atop the receiver chamber separated by the membrane in between, and horizontal diffusion cells where the donor and receiver chamber are arranged side-by-side. A version of the vertical diffusion cell model using Franz diffusion cells can be used for direct in vitro measurement of transdermal permeability. See, e.g., Ng, S.-F. et al. AAPS PharmSciTech 2010 Sep 15; 11(3): 1432-1441. Instruments and materials for Franz diffusion cell transdermal measurement are commercially available, e.g., from Logan Instruments (Somerset, NJ USA), Atlantis Bioscience (Singapore), and MilliporeSigma (a division of Merck KGaA, Darmstadt, Germany). Synthetic polymeric material, such as cellulose acetate or Strat-M®, can be used in membranes in Franz cell methods.

[0105] In some embodiments, a composition of the present disclosure delivers L-arginine across an in vitro membrane. L-arginine can be delivered in any amount, e.g., from about 0.01 mg to about 100 mg, over any period of time, e.g., from about 1 min to about 1 day. In some embodiments, a composition of the present disclosure delivers from about 0.01 mg to about 100 mg, e.g., from about 0.01 mg to about 50 mg, from about 0.01 mg to about 40 mg, from about 0.01 mg to about 30 mg, from about 0.01 mg to about 20 mg, from about 0.01 mg to about 10 mg, from about 0.1 mg to about 50 mg, from about 0.1 mg to about 40 mg, from about 0.1 mg to about 30 mg, from about 0.1 mg to about 20 mg, or from about 0.1 mg to about 10 mg, L-arginine across an in vitro membrane. In some embodiments, a composition of the present disclosure delivers L-arginine across a membrane in an in vitro membrane over from about 0.1 to about 12 hours, e.g., about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or about 12 hours. In some embodiments, a composition of the present disclosure delivers L-arginine across an in vitro membrane over about 2 hours. In some embodiments, a composition of the present disclosure delivers L-arginine across an in vitro membrane over about 8 hours. In some embodiments, a composition of the present disclosure delivers from about 0.1 mg to about 10 mg L-arginine across an in vitro membrane over about 2 hours. In some embodiments, a composition of the present disclosure delivers from about 0.1 mg to about 10 mg L-arginine across an in vitroAttorney Docket No. 063594-504001WOmembrane over about 8 hours. In some embodiments, a composition of the present disclosure delivers from about 1 mg to about 10 mg L-arginine across an in vitro membrane over about 2 hours. In some embodiments, a composition of the present disclosure delivers from about 1 mg to about 10 mg L-arginine across an in vitro membrane over about 8 hours. In some embodiments, a composition of the present disclosure delivers about 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1, 2, 3, 4, 5, 6, 7, 8, 9, or about 10 mg L-arginine across an in vitro membrane over about 8 hours.

[0106] In some embodiments, a composition of the present disclosure delivers L-arginine across a membrane in a Franz cell. Illustrative compositions of the present disclosure have shown measurable in vitro permeability, e.g., as described in Example 2 herein. L-arginine can be delivered in any amount, e.g., from about 0.01 mg to about 100 mg, over any period of time, e.g., from about 1 min to about 1 day. In some embodiments, a composition of the present disclosure delivers from about 0.01 mg to about 100 mg, e.g., from about 0.01 mg to about 50 mg, from about 0.01 mg to about 40 mg, from about 0.01 mg to about 30 mg, from about 0.01 mg to about 20 mg, from about 0.01 mg to about 10 mg, from about 0.1 mg to about 50 mg, from about 0.1 mg to about 40 mg, from about 0.1 mg to about 30 mg, from about 0.1 mg to about 20 mg, or from about 0.1 mg to about 10 mg, L-arginine across a membrane in a Franz cell. In some embodiments, a composition of the present disclosure delivers L-arginine across a membrane in a Franz cell over from about 0.1 to about 12 hours, e.g., about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, or about 12 hours. In some embodiments, a composition of the present disclosure delivers L-arginine across a membrane in a Franz cell over about 2 hours. In some embodiments, a composition of the present disclosure delivers L-arginine across a membrane in a Franz cell over about 8 hours. In some embodiments, a composition of the present disclosure delivers from about 0.1 mg to about 10 mg L-arginine across a membrane in a Franz cell over about 2 hours. In some embodiments, a composition of the present disclosure delivers from about 0.1 mg to about 10 mg L-arginine across a membrane in a Franz cell over about 8 hours. In some embodiments, a composition of the present disclosure delivers from about 1 mg to about 10 mg L-arginine across a membrane in a Franz cell over about 2 hours. In some embodiments, a composition of the present disclosure delivers from about 1 mg to about 10 mg L-arginine across a membrane in a Franz cell over about 8 hours. In some embodiments, a composition of the present disclosureAttorney Docket No. 063594-504001WOdelivers about 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1, 2, 3, 4, 5, 6, 7, 8, 9, or about 10 mg L-arginine across a membrane in a Franz cell over about 8 hours.

[0107] In some embodiments, a composition of the present disclosure delivers more L-arginine transdermally as compared to a literature L-arginine composition. The amount of L-arginine can be determined by any in vitro or in vivo method in the art. In some embodiments, a composition of the present disclosure delivers from about 1.1 times to about 1,000,000 times, e.g., from about 2 times to about 100,000 times, from about 10 times to about 100,000 times, from about 100 times to about 100,000 times, from about 10 times to about 10,000 times, from about 100 times to about 10,000 times, from about 10 times to about 1,000 times, from about 100 times to about 1,000 times, from about 2 times to about 100 times, from about 10 times to about 100 times, or from about 1.1 to about 10 times, more L-arginine transdermally as compared to a literature L-arginine composition. In some embodiments, a composition of the present disclosure delivers about 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, 2, 3, 4, 5, 6, 7, 8, 9, 10, 20, 30, 40, 50, 60, 70, 80, 90, or about 100 times more L-arginine transdermally as compared to a literature L-arginine composition.B. Therapeutic Methods and / or Uses

[0108] The present disclosure provides methods and uses for treating decreased blood flow, e.g., poor circulation, e.g., associated with diabetes, comprising topically administering L-arginine compositions. The methods and uses described herein provide an effective amount of the L-arginine while reducing the incidence of one or more of the adverse effects observed with treatment by L-arginine administration by other means, such as intravenous, intramuscular, or oral administration.

[0109] Methods and uses of the present disclosure include locally increasing blood flow, e.g., at a specific part of the body, in a human subject in need thereof, comprising topically administering to the subject the composition of the present disclosure. In some embodiments, the method and / or use comprises treating a disease or condition that benefits from locally increasing blood flow.

[0110] In some embodiments, a method of the present disclosure is a method of enhancing, e.g., increasing, blood flow in a human subject in need thereof, comprising topicallyAttorney Docket No. 063594-504001WOadministering to the subject the composition of the present disclosure. Tn some embodiments, the method comprises administering the composition of the present disclosure on the extremities, e.g., arms, legs, and / or feet. The human subject can be a diabetes patient. In some embodiments, the method comprises administering the composition of the present disclosure on the extremities, e.g., arms, legs, and / or feet, of a diabetes patient. In some embodiments, the method of the present disclosure is a method of enhancing blood flow in an extremity in a human subject in need thereof, comprising topically administering to the extremity a composition as described herein. Accordingly, in some embodiments, the administering can comprise administering on a leg and / or a foot. In some embodiments, the extremity comprises a lower leg. In some embodiments, the extremity comprises an arm.[OHl] In diabetes, the condition of hyperglycemia reduces nitric oxide (NO), which is important for the regulation of vasodilation, anticoagulation, the proliferation of smooth muscle, and the overall antioxidant capacity of endothelial cells. L-Arg also serves as a basic substrate to produce NO in endothelial cells, thus regulating vascular tone and overall cardiovascular homeostasis. The effect of L-Arg and tetrahydrobiopterin infusion was investigated on endothelial dysfunction induced by ischemia / reperfusion in patients with type 2 diabetes mellitus and coronary artery disease. L-Arg supplementation significantly attenuated endothelial dysfunction in this type of patients. Hence, an implementation of L-Arg could potentially reduce some of the main and most serious complications of diabetes, including heart failure; indeed, diabetic patients are more prone to develop cardiomyopathy than healthy subjects.

[0112] Oral L-arginine at 5 g daily for 14 days was reported to improve microcirculatory function in healthy young women and elderly women who were healthy or had type 2 diabetes mellitus. See, Costa, G. et al. J. Vase. Res (2022), 59 (1): 24-33.

[0113] L-Arginine is an amino acid that is commercially available in oral supplements.However, oral L-arginine may cause certain side effects. Oral use of L-arginine might cause: nausea, abdominal pain, diarrhea, bloating, gout, headache, allergic response, and / or airway inflammation or worsening of asthma symptoms. L-arginine is not recommended for people who have had a recent heart attack due to concerns that the supplement might increase the risk of death. L-arginine can worsen allergies or asthma. Excessive systemic L-arginine can also trigger the virus that causes cold sores or genital herpes.Attorney Docket No. 063594-504001WO

[0114] Methods and uses of the present disclosure include increasing blood flow in a human subject suffering from or suspected of suffering from chemotherapy-induced neuropathy, also known as chemotherapy induced peripheral neuropathy (CIPN). Accordingly, in some embodiments, the human subject is a cancer patient. Chemotherapy-induced peripheral neuropathy (CIPN) is a side effect caused by one or more antineoplastic agents. Clinically, CIPN is a mostly sensory neuropathy that may be accompanied by motor and autonomic changes of varying intensity and duration. Currently, there is no single effective method of preventing CIPN; moreover, the possibilities of treating this syndrome are very limited. CIPN can be caused by platinum-based antineoplastic agents, vinca alkaloids, epothilones (e.g., ixabepilone), taxanes, proteasome inhibitors (e.g., bortezomib) and immunomodulatory drugs (e.g., thalidomide).Among them, the most neurotoxic are platinum-based agents, taxanes, ixabepilone and thalidomide; other less neurotoxic but also commonly used drugs are bortezomib and vinca alkaloids.

[0115] Oral L-arginine has been investigated in phase II clinical trials for prevention of bortezomib-induced peripheral neuropathy in newly diagnosed multiple myeloma patients. See, Maschio, M. et al. Integrative Cancer Therapies 2022, 21, pages 1-10. However, the treatment was not adequate to prevent peripheral neuropathy.

[0116] Methods and techniques for measuring blood flow are known in the art, including but are not limited to temperature measurements, e.g., infrared thermography or thermometers such as a temporal thermometer, blood pressure measurement, and Doppler ultrasound. For instance, Doppler ultrasound is a technique that uses high-frequency sound waves to measure the amount of blood flow through a subject’s arteries and veins, e.g., in arms and legs. An increased blood flow can be determined by comparing blood flow after treatment to a control, such as untreated extremity or baseline before treatment. In some embodiments, the blood flow is measured using Doppler ultrasound. In some embodiments, the blood flow is measured at an artery or a vein. In some embodiments, the artery is the dorsalis pedis artery. In some embodiments, the artery is the posterior tibialis artery.

[0117] In some embodiments, blood flow can be assessed by determination of erythema, that is, the superficial reddening of capillaries in the skin. In some embodiments, erythema is measured by colorimetry. Colorimeters are small, handheld devices used to obtain objectiveAttorney Docket No. 063594-504001WOmeasurements of color using a three-dimensional color space via the international standard Commission Internationale de 1’Eclairage system, which is represented by three axes: L*, a*, and b*. The L* axis is a grayscale with values from 0 (black) to 100 (white). The L* value correlates with the level of skin pigmentation. The a* axis describes red and green values, which correlates with erythema. The b* axis describes yellow and blue values and correlates with pigmentation and tanning. Commercial colorimeters, for instance, ColorMeter DSM III (ColorMeter; Cortex Technology) are available.

[0118] In some embodiments, the method or use of the present disclosure comprises enhancing, e.g., increasing, blood flow in a subject, e.g, in an extremity such as a leg in a subject, by from about 10% to about 1000%, such as from about 20% to about 900%, from about 20% to about 800%, from about 20% to about 700%, from about 20% to about 600%, from about 20% to about 500%, from about 20% to about 400%, from about 20% to about 300%, from about 20% to about 200%, from about 20% to about 100%, from about 20% to about 90%, from about 20% to about 80%, from about 20% to about 70%, from about 20% to about 60%, from about 20% to about 50%, from about 20% to about 40%, from about 20% to about 30%, from about 30% to about 80%, from about 30% to about 70%, from about 30% to about 60%, or from about 30% to about 50% as compared to blood flow in a control. In some embodiments, the method or use of the present disclosure increased blood flow in a subject by about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 200%, 300%, 400%, 500%, 600%, 700%, 800%, 900%, or about 1000% as compared to a control. In some embodiments, the control is an untreated extremity in the subject. In some embodiments, the control is the extremity in the subject before treatment by the composition of the present disclosure.

[0119] In some embodiments, the method or use of the present disclosure reduces the incidence and / or severity of one or more of the adverse effects in a subject by from about 10% to about 90%, such as from about 20% to about 90%, from about 20% to about 80%, from about 20% to about 70%, from about 20% to about 60%, from about 20% to about 50%, from about 20% to about 40%, from about 30% to about 90%, from about 30% to about 80%, from about 30% to about 70%, from about 30% to about 60%, or from about 30% to about 50% as compared to the incidence and / or severity of one or more of the adverse effects in the subject treated with oral L-arginine. In some embodiments, the method or use of the present disclosure reduces theAttorney Docket No. 063594-504001WOincidence and / or severity of one or more of the adverse effects in a subject by about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, or about 90% as compared to the incidence and / or severity of one or more of the adverse effects in the subject treated with oral L-arginine.

[0120] In some embodiments, the method or use of the present disclosure reduces the probability of one or more of the adverse effects in a subject by from about 10% to about 90%, such as from about 20% to about 90%, from about 20% to about 80%, from about 20% to about 70%, from about 20% to about 60%, from about 20% to about 50%, from about 20% to about 40%, from about 30% to about 90%, from about 30% to about 80%, from about 30% to about 70%, from about 30% to about 60%, or from about 30% to about 50% as compared to the probability of one or more of the adverse effects in the subject treated with oral L-arginine. In some embodiments, the method or use of the present disclosure reduces the probability of one or more of the adverse effects in a subject by about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, or about 90% as compared to the probability of one or more of the adverse effects in the subject treated with oral L-arginine.

[0121] In some embodiments, the method comprises topically administering once, twice, or three times daily. In some embodiments, the method comprises topically administering three times daily. When administered multiple times, the time period between adminstration can be the same or different. In some embodiments, the time period between administration is the same, e.g., every 12 hours for two times daily. In some embodiments, the time period between administration is different, e.g., at 0, 4, and 8 hours three times on a daily basis. In some embodiments, the method comprises topically administering about every four hours.

[0122] In some embodiments, the method comprises administering for at least about 8 days.

[0123] In some embodiments, the method comprises administering about 5 g of the composition of the present disclosure per dose. In some embodiments, the dose is the amount of composition administered each time.

[0124] Because the compositions of the present disclosure are believed to afford a safer toxicity profile, the methods and uses described herein can be used to treat patient populations that are contraindicated or are not recommended for use with oral L-arginine methods. For instance, oral L-arginine can interact with other drugs. Possible interactions include:Attorney Docket No. 063594-504001WO• Anticoagulants and anti-platelet drugs, herbs and supplements. These types of drugs, herbs and supplements reduce blood clotting. Taking L-arginine with them might increase the risk of bleeding.• Blood pressure drugs. herbs and supplements. L-arginine might lower blood pressure in people who have high blood pressure. Combining use of L-arginine with a blood pressure drug, herb or supplement might increase the risk of blood pressure becoming too low. • Diabetes drugs, herbs and supplements. L-arginine might decrease blood sugar levels in people with diabetes. Dosage of diabetes drugs, herbs or supplements may need to be adjusted.• Isoproterenol. Use of this heart medication with L-arginine might cause blood pressure to become too low.• Nitrates. Use of this chest pain medication with L-arginine might cause blood pressure to become too low.• Water pills (potassium-sparing diuretics!. L-arginine is not recommended to be taken with diuretics such as amiloride, spironolactone, or triamterene. These medications can increase potassium levels, increasing the risk of developing a higher than normal level of potassium in your blood (hyperkalemia).• Sildenafil. Use of the erectile dysfunction medication with L-arginine might cause blood pressure to become too low.

[0125] The dosage or dosing frequency of a composition of the present disclosure may be adjusted over the course of the treatment.

[0126] The compounds and compositions of the present disclosure can be co-administered with other agents. Co-administration includes administering the compound or composition of the present disclosure within 0.5, 1, 2, 4, 6, 8, 10, 12, 16, 20, or 24 hours of the other agent. Coadministration also includes administering simultaneously, approximately simultaneously (e.g., within about 1, 5, 10, 15, 20, or 30 minutes of each other), or sequentially in any order.Moreover, the compounds and compositions of the present disclosure can each be administeredAttorney Docket No. 063594-504001WOonce a day, or two, three, or more times per day so as to provide the preferred dosage level per day.

[0127] In some embodiments, co-administration can be accomplished by co-formulation, that is, preparing a single pharmaceutical composition including the compounds and compositions of the present disclosure and any other agent. Alternatively, the various components can be formulated separately.

[0128] In some embodiments, a use of the present disclosure comprises a composition of the present disclosure for the manufacture of a medicament for aiding in the treatment of inflammation or pain in a subject in need thereof. In some embodiments, the use comprises a composition as described in Section IV above.

[0129] In some embodiments, a composition of the present disclosure is for use in aiding in the enhancement of blood flow in a subject in need thereof. In some embodiments, the compound for use comprises a pharmaceutical composition as described in Section IV above.

[0130] In some embodiments, a use of the present disclosure comprises a composition of the present disclosure for the manufacture of a medicament for enhancing blood flow in a subject in need thereof. In some embodiments, the use comprises a composition as described in Section IV above.

[0131] In some embodiments, a composition of the present disclosure is for use in enhancing blood flow in a subject in need thereof. In some embodiments, the composition for use is as described in Section IV above.

[0132] Kits that comprise a composition of the present disclosure are also included in the present disclosure. In some embodiments, a kit further includes instructions for use. In some embodiments, a kit includes a composition of the disclosure, and a label and / or instructions for use of the composition in the treatment of the indications, such as the diseases or conditions, described herein. In some embodiments, kits comprising a composition of the present disclosure in combination with one or more (e.g., one, two, three, four, one or two, or one to three, or one to four) additional therapeutic agents are provided.Attorney Docket No. 063594-504001WO

[0133] Provided herein are also articles of manufacture that include a composition of the present disclosure in a suitable container. In some embodiments, the container is a vial, jar, tube, or ampoule.VI. EXAMPLES

[0134] The following examples are provided to further aid in understanding the embodiments disclosed in the application, and presuppose an understanding of conventional methods well known to those persons having ordinary skill in the art to which the examples pertain. The particular materials and conditions described hereunder are intended to exemplify particular aspects of embodiments disclosed herein and should not be construed to limit the reasonable scope thereof.Example 1. L-Arginine Composition Preparations

[0135] Exemplary compositions of the present disclosure were prepared according to the methods described herein.Preparation of Composition Fl 40-QQ8EtOH

[0136] The following procedure was used to prepare Composition F140-008EtOH at 50 g batch size.Table 1. Components and Amounts of CompositionAttorney Docket No. 063594-504001WO

[0137] Intermediate mixtures containing the components in Table 1 were prepared by combining the ingredients at the appropriate stirring conditions and temperature as described in Table 2.Table 2. Stirring and Heating Conditions for Intermediate Mixtures

[0138] Intermediate Mixture A was prepared last. Alternatively, Intermediate Mixture A was stirred at 200-300 rpm at room temperature until all other Intermediate Mixtures were prepared at the temperature shown in the table. Optionally, L-arginine particle size was reduced by grinding in a mortar and pestle for 5 minutes before preparation of Intermediate Mixture A.

[0139] Intermediate Mixture B was slowly poured into Intermediate Mixture A with stirring. The resulting mixture was stirred to break apart any material that may have agglomerated at the bottom or on the sides of the container. This resulting mixture was then heated to 80 °C to boil off ethanol, whereupon the viscosity of the mixture increased.Attorney Docket No. 063594-504001WO

[0140] Intermediate Mixture C at 70 °C was then added to the above mixture at 80 °C with stirring. Using a metal spatula, the mixture was gently stirred to ensure that Intermediate Mixture A+B+C was thoroughly combined.

[0141] When Intermediate Mixture D was at 65 °C, and Intermediate Mixture A+B+C was at least 65°C, Intermediate Mixture D was poured into Intermediate Mixture A+B+C while stirring. Using a metal spatula, the resulting composition was stirred to ensure that thorough mixing. The temperature was increased to 95 °C while stirring at 200-300 rpm. The remaining L-arginine dissolved to form a yellow-orange solution. The mixture was then removed from the heat with continued stirring, followed by incubation at room temperature overnight.Preparation of Composition F140-QQ9EtOH

[0142] The following procedure was used to prepare Composition F140-009EtOH at 50 g batch size.Table 3. Components and Amounts of CompositionAttorney Docket No. 063594-504001WOTable 4. Stirring and Heating Conditions for Intermediate Mixtures

[0143] The procedure for combining Intermediate Mixtures A, B, C, and D was the same as described for Composition F140-008EtOH.Preparation of Composition F140-009IPA

[0144] The following procedure was used to prepare Composition F140-009IPA at 10 g and 650 g batch size.Table 5. Components and Amounts of CompositionAttorney Docket No. 063594-504001WOTable 6. Stirring and Heating Conditions for Intermediate Mixtures

[0145] The procedure for combining Intermediate Mixtures A, B, C, and D was the same as described for Composition F140-008EtOH, with the exception of the ingredients listed in Table 5 and the temperatures noted in Table 6.

[0146] The following compositions were prepared according to the procedures described above, with percent weight amounts listed in Table 7 through Table 10 below.Table 7. L-Arginine CompositionsAttorney Docket No. 063594-504001WOTable 8. L-Arginine CompositionsAttorney Docket No. 063594-504001WOaalso contained 10% myristic acid;balso contained 4.84% menthol.Table 9. L-Arginine CompositionsAttorney Docket No. 063594-504001WOaF140004 also contained 4% capric acid, 1% propanoic acid;bF120005 also contained 2% capric acid, 4% myristic acid, and 5% stearic acid.Table 10. L-Arginine CompositionsAttorney Docket No. 063594-504001WOc140-001-lHMK also contained 2% caffeine, 4% capric acid, 2.7% propionic acid, 1.5% rosemary oil

[0147] Composition F140-009IPA was substantially the same as described for Composition F140-009EtOH above, except that isopropanol was used in its preparation instead of ethanol.Example 2. In vitro Studies on L-Arginine Composition Permeability

[0148] The compositions prepared in Example 1 were tested for in vitro permeability in Logan Franz cell protocol. The Logan Instrument Automated Franz Cell was utilized for this experiment with receptor fluid containing 90 (IX isotonic phosphate buffered saline at pH 5.0): 10 (Acetonitrile) ratio. Strat M Membranes were then fitted to the receptor compartments and secured with a dosing disk pressed into a donor compartment and a clamp. The cells were then all filled with 12mL of receptor fluid and about 2.0 g of formulation was transferred to the donor compartment using a spatula, weight of any residual formulation on either spatula or weigh boat was recorded . The Logan Instrument then took ImL samples at 7 min, 1 h, 2 h, 3 h, 4 h, 5 h, 6 h, 7 h and 8 h. The 2 h and 8 h time points were used for cross-formulation performance comparisons.

[0149] HPLC Instrument Protocol: The HPLC was dry primed, wet primed, and the injector was purged before equilibrating and cooling the column to 22° C using the L-arginine isocratic HPLC method. This method uses a degassed buffer containing 25:75 5.0 mM potassium dihydrogen phosphate at pH 2.95 : acetonitrile at a flow rate of ImL / min. Each sample runs for 10.0 minutes with the L-arginine peak occurring at about 7.8 - 8.1 minutes. Injection volume: 20 pL. A standard curve was established and the concentration of samples was calculated using the equation established by the standard. Each sample from the Logan machine was run using the HPLC protocol and a concentration vs time curve was generated. In order to better matchAttorney Docket No. 063594-504001WOionization state of the L-arginine in the receptor fluid to the ionization state of the L-arginine in the mobile phase, 0.5 mb of acetonitrile was added to both sample and standard solutions.Results are summarized in Table 11 through Table 14 below.

[0150] A comparative Healthy Feet formulation containing 13% L-arginine hydrochloride, 2% oleic acid, 15% glyceryl monostearate, 2% polysorbate 20, 2% isopropyl myristate, 2% menthol, 5% choline chloride, 5% magnesium chloride, 5% potassium chloride, and 10% sodium chloride demonstrated average cumulative transfer of 0 mg at 2 h and at 8 h.Table 11. PermeabilityTable 12. PermeabilityTable 13. PermeabilityAttorney Docket No. 063594-504001WOTable 14. Permeability

[0151] The present compositions exhibited measurable transfer of L-arginine across a membrane in the in vitro studies. Adding 4.8% (w / w) menthol (Composition F13m) significantly attenuated the transfer of L-arginine across the membrane. Significant levels of dimethyl sulfoxide (Compositions F8, F9, Fl OB) afforded compositions that were capable of delivering significant levels of L-arginine across the membrane after 8 h. Nevertheless, certain compositions, e.g., Composition Fl 6, with less than about 5% dimethyl sulfoxide, did demonstrate significant L-arginine transfer after 8 h.Example 3. L-Arginine Composition Effects in Healthy Volunteers with Reduced Circulation in Extremities

[0152] Exemplary Composition F140-009IPA was topically administered in a proof-of-concept label claim study in healthy volunteers demonstrating skin signs of vascular insufficiency. This study was aimed at examining a novel topical formulation for its ability to improve the appearance of the lower legs and feet.

[0153] Test Product: Composition F140-009IPA (5 g, the “study cream”) was administered at 8 am, noon, and 4 pm each treatment day for 8 days total. Dose application site was lower leg, from the mid-calf to the top of the foot in a stocking distribution. On the clinic visit days, the dose application area of both legs was cleaned and dried prior to each blinded Doppler measurements. Dose administration was by the subject under research staff supervision. Gloves were worn during study cream application. Study cream was applied to clean, dry skin. Study cream was rubbed into entire application site for approximately 5 minutes. Dose application site was left undisturbed for 15 minutes after application, after which the residual cream was removed with dry paper towel.Attorney Docket No. 063594-504001WO

[0154] Study population: otherwise healthy males and females demonstrating skin signs of vascular insufficiency due to decreased arterial circulation. Ten subjects total were enrolled. Endpoints:

[0155] Tolerability Endpoint: The tolerability endpoint was the investigator-assessed absence of skin irritation from the study product at any time during the 1-week study.

[0156] Safety Endpoint: The safety endpoint was the overall incidence of all adverse events reported during the study.

[0157] Co-Primary Efficacy Endpoints: one co-primary efficacy endpoint was a statistically significant increase in the Doppler ultrasound measured blood flow in the dorsalis pedis artery and the posterior tibial artery after 1 week of treatment with the study product as compared to baseline.

[0158] One co-primary efficacy endpoint was a statistically significant increase in the Doppler ultrasound measured blood flow in the dorsalis pedis artery and the posterior tibial artery after 1 week of treatment of the randomized target leg as compared to the opposite untreated leg at all time points.

[0159] Secondary Efficacy Endpoints: one secondary efficacy endpoint was a statistically significant increase in skin temperature measured by temporal thermometer in the target site selected at baseline as compared to baseline.

[0160] One secondary efficacy endpoint was a statistically significant increase in skin temperature measured by temporal thermometer in the target site selected at baseline as compared to the opposite untreated leg at all time points.

[0161] One secondary efficacy endpoint was a statistically significant shift in color from blue to red tones measured by colorimeter in the target site selected at baseline as compared to baseline.

[0162] One secondary efficacy endpoint was a statistically significant shift in color from blue to red tones measured by colorimeter in the target site selected at baseline as compared to the opposite untreated leg at all time points.Attorney Docket No. 063594-504001WOMeasures:

[0163] Investigator Assessed Tolerability: redness, swelling, and dryness. All assessments were made on a 5-point ordinal scale (0=none, l=minimal, 2=mild, 3=moderate, 4=severe) at baseline and week 1.

[0164] Subject Assessed Tolerability: burning, stinging, tingling, and itching. All assessments were made on a 5-point ordinal scale (0=none, l=minimal, 2=mild, 3=moderate, 4=severe) at baseline and week 1.

[0165] Noninvasive Assessments: Doppler ultrasound (equipment provided by sponsor) measurements were taken over the dorsalis pedis artery and the posterior tibial artery at baseline, noon, and 4pm prior to study product application on the days the subjects are domiciled (Monday, Thursday, Monday). Approximately one hour prior to the end-of-day Doppler ultrasound measurements of posterior tibial artery and dorsal pedis artery blood flow on each study visit day, the dose application site was cleaned with gentle soap / detergent wipes to remove any remaining study cream. Temporal thermometer measurements of skin temperature were taken of both legs at baseline, noon, and 4pm prior to study product application on the days the subjects are domiciled (Monday, Thursday, Monday). Colorimeter measurements were taken of both legs at baseline, noon, and 4pm prior to study application on the days the subjects are domiciled (Monday, Thursday, Monday).Statistical Methods:

[0166] Along with descriptive statistics (means, standard deviations and percentages), Doppler ultrasound and skin temperature numeric data were analyzed using a Student t test with all time points compared to baseline. Change was considered significant at the alpha level of 0.05.Investigator and subject ordinal nonparametric results were analyzed using Wilcoxon signed rank test and sign test for paired comparison at different time points. Significance was defined at the p<0.05 level based on a two-sided test.Study Design Overview

[0167] Subjects who signed consent and met all inclusion criteria and none of the exclusion criteria were enrolled at the baseline visit. Subjects were screened to confirm that their loss ofAttorney Docket No. 063594-504001WOlower extremity aesthetic was due to their reduced leg circulation as assessed by the dermatologist investigator. Subjects were domiciled for the entire day Monday from 8am until 4pm. A target site was selected and marked for each measurement to be used throughout the study. Subjects underwent an investigator performed Doppler ultrasound of both legs at baseline over the dorsalis pedis artery and the posterior tibial artery prior to 8am on Monday. The subject, under supervision of the research staff, applied the study cream to one randomized leg from the mid-calf to the top of the foot, avoiding the sole of the foot, in a stocking distribution. The opposite leg received no treatment. Subjects could move around the office without exertion. The Investigator, who was blinded to treatment leg, performed a second Doppler ultrasound of both legs over the dorsalis pedis artery and the posterior tibial artery at noon. A temporal thermometer was used to determine the skin temperature of both legs. A colorimeter was used to determine the erythema of both legs. After these measurements were completed, a second application of the cream to the same area on the randomized treatment leg was administered. Again, the opposite leg received no treatment. A third blinded Investigator-performed Doppler ultrasound examination of the dorsalis pedis artery and the posterior tibial artery of both legs (treated and untreated) occurred at 4 pm. A temporal thermometer was used to determine the skin temperature of both legs. A third application by the subject, under the supervision of the research staff, of the cream to the target area on the treated leg was administered with no product application to the opposite leg.

[0168] Subjects were discharged home with cream to continue the application schedule of 8am, noon, and 4pm on the leg selected for treatment. Text reminders were sent to encourage compliance along with a compliance diary. Subjects self-applied the study cream at home on Tuesday and Wednesday. Subjects returned to the research center on Thursday.

[0169] Subjects returned to the research center on Thursday and domiciled for the entire day from 8am until 4pm. Instrument measurements of both legs and study cream administration were performed according to the protocol described above for the initial Monday.

[0170] Subjects self-applied the study cream at home on Friday, Saturday, and Sunday.Subjects returned to the research center on Monday and domiciled for the entire day from 8am until 4pm. Instrument measurements of both legs and study cream administration were performed according to the protocol described above for the initial Monday.Attorney Docket No. 063594-504001WOResults

[0171] No skin irritation or any safety issues were observed.

[0172] Circulation improved in 8 out of 10 participants after 8 days of treatment with 5 g Composition F140-009IPA three times daily, shown by the change from baseline in the percent relative difference in blood flow between treated and untreated legs at the posterior tibialis artery. Subjects in the treated group showed an improvement from daily baseline in blood flow rate in posterior tibalis artery when compared to the untreated group. See, FIG. 1. Subjects in the treated group showed an overall improved blood flow rate from baseline in posterior tibalis artery over the course of the study when compared to the untreated group. See, FIG. 2. A statistically significant 115% increase from baseline was observed in average blood flow in the treated leg versus untreated leg at end of Day 8, p-value 0.039. See, FIG. 3.

[0173] Subjects in the treated group showed an improvement from daily baseline in blood flow rate in dorsalis pedis artery when compared to the untreated group. See, FIG. 4. Subjects in the treated group showed an overall improved blood flow rate from baseline in dorsalis pedis artery over the course of the study when compared to the untreated group. See, FIG. 5.

[0174] Circulation improvement was shown by the change from baseline in the percent relative difference in blood flow between treated and untreated legs as measured by Doppler ultrasound:8 out of 10 participants showed improvement based on measurements at the posterior tibialis artery. Mean difference between treated and untreated leg was 26% on Day 8, achieving statistical significance with p-value of 0.016.6 out of 10 participants showed improvement based on measurements at the dorsalis pedis artery;- 2 participants showed decreased relative blood flow at both arteries; and2 participants showed increased relative blood flow at the posterior tibialis artery and decreased relative blood flow at the dorsalis pedis artery.

[0175] Investigator reported that the posterior tibialis artery was easier to find and measure blood flow.Attorney Docket No. 063594-504001WO

[0176] Temperature measurement as used in this study may not be sensitive enough to determine changes in temperature resulting from difference in blood flow. The treated and untreated groups yielded statistically similar results.

[0177] Erythema measurement as used in this study may not be sensitive enough to determine changes in erythema resulting from difference in blood flow. The treated and untreated groups yielded statistically similar results.

[0178] Although the foregoing invention has been described in some detail by way of illustration and Example for purposes of clarity of understanding, one of skill in the art will appreciate that certain changes and modifications may be practiced within the scope of the appended claims. In addition, each reference provided herein is incorporated by reference in its entirety to the same extent as if each reference was individually incorporated by reference. Where a conflict exists between the instant application and a reference provided herein, the instant application shall dominate.

Claims

WHAT IS CLAIMED IS:

1. A composition comprising:(a) from about 1% to about 50% (w / w) L-arginine; and(b) from about 3% to about 90% (w / w) oleic acid;wherein the L-arginine and the oleic acid form a mixture having a lower melting point than pure oleic acid.

2. The composition of claim 1, comprising from about 3% to about 40% (w / w) L-arginine.

3. The composition of any one of claims 1 to 2, comprising from about 5% to about 50% (w / w) oleic acid.

4. The composition of any one of claims 1 to 3, comprising from about 2% to about 20% (w / w) caprylic acid.

5. The composition of any one of claims 1 to 4, comprising from about 2% to about 20% (w / w) lauric acid.

6. The composition of any one of claims 1 to 5, comprising from about 2% to about 20% (w / w) levulinic acid.

7. The composition of any one of claims 1 to 6, comprising from about 5% to about 20% (w / w) glyceryl monostearate.

8. The composition of any one of claims 1 to 7, comprising from about 10% to about 30% (w / w) isopropyl myristate.

9. The composition of any one of claims 1 to 8, comprising from about 2% to about 20% (w / w) squalane.

10. The composition of any one of claims 1 to 9, comprising less than about 5% (w / w) dimethyl sulfoxide.

11. The composition of any one of claims 1 to 10, comprising from about 1% to about 2% (w / w) eucalyptol.

12. The composition of any one of claims 1 to 11, which is substantially free of water.

13. The composition of any one of claims 1 to 12, comprising:(a) from about 10% to about 15% (w / w) L-arginine;(b) from about 10% to about 30% (w / w) oleic acid;(c) from about 2% to about 10% (w / w) caprylic acid;(d) from about 2% to about 20% (w / w) lauric acid;(e) from about 8% to about 16% (w / w) levulinic acid;(f) from about 7% to about 15% (w / w) glyceryl monostearate;(g) from about 10% to about 30% (w / w) isopropyl myristate;(h) from about 2% to about 20% (w / w) squalane;(i) from about 0.5% to about 1% (w / w) dimethyl sulfoxide; and(j) from about 1% to about 2% (w / w) eucalyptol;wherein the L-arginine and the oleic acid form a mixture having a lower melting point than pure oleic acid.

14. A method of enhancing blood flow in a human subject in need thereof, comprising topically administering to the subject the composition of any one of claims 1 to 13.

15. The method of claim 14, wherein the human subject is a diabetes patient.

16. The method of claim 14, wherein the administering comprises administering on a leg and / or a foot.

17. The method of any one of claims 14 to 16, comprising topically administering once, twice, or three times daily.

18. A method of enhancing blood flow in an extremity in a human subject in need thereof, comprising topically administering to the extremity the composition of any one of claims 1 to 13.

19. The method of claim 18, wherein the composition comprises:(a) about 14% (w / w) L-arginine;(b) about 19% (w / w) oleic acid;(c) about 6.1% (w / w) caprylic acid;(d) about 7.5% (w / w) lauric acid;(e) about 10% (w / w) levulinic acid;(f) about 14% (w / w) glyceryl monostearate;(g) about 20% (w / w) isopropyl myristate;(h) about 7% (w / w) squalane;(i) about 1% (w / w) dimethyl sulfoxide; and(j) about 1.4% (w / w) eucalyptol;wherein the L-arginine and the oleic acid form a mixture having a lower melting point than pure oleic acid.

20. The method of claim 18, comprising administering about 5 g of the composition.

21. The method of claim 18, comprising administering three times daily.

22. The method of claim 21, comprising administering about every four hours.

23. The method of claim 21, comprising administering for at least about eight days.

24. The method of claim 18, wherein the extremity comprises a lower leg.

25. The method of claim 18, wherein the enhanced blood flow is compared to an untreated extremity.

26. The method of claim 18, comprising increasing blood flow by from about 10% to about 1000% compared to blood flow in an untreated extremity.

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