Transdermal insulin preparation and method of using the same

Abstract

Describe a pharmaceutical preparation for the transdermal administration of insulin by topical application to the skin of humans or other animals. A methodology for formulating such a preparation that provides very rapid uptake of insulin, as well as migration to the skin and transdermal migration to either adipose tissue or the vasculature, while minimizing skin irritation and / or immune response, is based on a transdermal delivery system in which insulin forms a true solution in the form of a complex formed from a specific solvent in combination with a skin stabilizer and solvent and solute modifiers. Uptake of the therapeutic agent is further facilitated and made more rapid by including forskolin or other cellular energy sources, i.e., by induction of cAMP or cGMP.

Description

Technical Field

[0001] Related Applications This application claims priority based on U.S. Provisional Patent Application No. 63 / 346,022, filed on May 26, 2020. The entire disclosure of the priority application is incorporated herein by reference.

[0002] The present disclosure relates to transdermal insulin formulations, as well as methods of administering and preparing such formulations.

Background Art

[0003] Insulin is a natural hormone secreted by pancreatic islet beta cells in response to an increase in blood glucose levels. This hormone acts to control processes involved in glucose metabolism and the intermediate metabolism of fats, carbohydrates, and proteins. Insulin lowers blood glucose levels and promotes the transport and uptake of glucose into muscle cells and other tissues. Due to the chemical nature of the insulin molecule, the delivery route of insulin administration in diabetic patients who require multiple daily insulin administrations is intradermal or subcutaneous injection.

[0004] The development efforts of non-injectable transdermal insulin delivery systems for the treatment of diabetes have not been successful to date.

[0005] Attempts have been made to develop transdermal "patches" or external pumps containing a specific amount of insulin that migrates at a specific rate, but these patches and pumps have numerous limitations. One specific limitation is that insulin users often have to accurately estimate their required amounts in relation to physical activity and carbohydrate intake. Additionally, there are different types of insulin, such as long-acting and short-acting, and patients have to become familiar with mixing both the type and amount of insulin to appropriately control their blood glucose levels. The use of multiple patches with different dosage strengths and insulin response characteristics is thus problematic.

[0006] Therefore, there has long been a pressing need for a convenient form of transdermal insulin delivery system and a better means of delivering insulin to patients in need thereof. SUMMARY OF THE INVENTION

[0007] The present application discloses a transdermal insulin formulation.

[0008] In one example, the formulation contains insulin and may be present in an amount in the range of 0.001% to 3.5% (wt / wt) of the total formulation amount. In one example, the insulin is rapid-acting insulin. In another example, the insulin is short-acting insulin. In another example, the insulin is intermediate-acting insulin. In another example, the insulin is long-acting insulin. In another example, the insulin may be one or more selected from the group consisting of rapid-acting insulin, short-acting insulin, intermediate-acting insulin, and long-acting insulin.

[0009] In another example, the formulation further contains a solvent system.

[0010] In one example, the solvent system contains two or more solvents.

[0011] In another example, the solvent system contains at least one solvent modifier.

[0012] In another example, the solvent system contains at least one solute modifier.

[0013] In another example, the solvent system contains at least one source of cell activation energy.

[0014] In another example, the solvent system contains at least one skin stabilizer.

[0015] In another example, the solvent system includes two or more solvents, at least one solvent modifier, at least one source of cell activation energy, and at least one skin stabilizer.

[0016] In another example, the solvent system includes one or more components selected from the group consisting of two or more solvents, at least one solvent modifier, at least one solute modifier, at least one source of cell activation energy, and at least one skin stabilizer.

[0017] The two or more solvents can be selected from the group consisting of ethanol, ethylene glycol, propylene glycol, propylene carbonate, butylene glycol, acetone, and glycerol; and can be present in an amount in the range of 80% - 99% (wt / wt) of the total formulation amount.

[0018] The at least one solvent modifier can be selected from the group consisting of lemon oil (or / and d-limonene), vitamin E, provitamin B, D-panthenol, and methylsulfonylmethane (MSM); and can be present in an amount in the range of 0.0001% - 20% (wt / wt) of the total formulation amount.

[0019] The at least one solute modifier can be selected from the group consisting of terpenes, oxindole alkaloids, quercitrin (a glycoside of quercetin), genistein and its glucosides, genistein, polyphenolic flavonoids, and other sugar adduct glucuronides; and can be present in an amount in the range of 0.003% - 5% (wt / wt) of the total formulation amount.

[0020] The at least one source of cell activation energy can be selected from the group consisting of forskolin, colforsin, methylxanthines, psychogenin and psychotrisaponin, angelic acid, feropterin, oxypoisidanin, acetylcholine, cytidine diphosphate choline, and ascorbic acid; and can be present in an amount in the range of 0.01% - 0.1% (wt / wt) of the total formulation amount.

[0021] At least one type of skin stabilizer can be selected from the group consisting of glycerin monolaurate, vitamin D3, alkoxyglycerols, eicosapentaenoic acid (EPA), docosahexaenoic acid (DHA), gamma-linolenic acid (GLA), vitamin E, D-panthenol, phytantriol, dehydroepiandrosterone (DHEA), pregnenolone, pregnenolone acetate, esculin, allantoin, and ascorbyl palmitate; and can be present in an amount in the range of 0.05% to 5% (wt / wt) of the total amount of the formulation.

[0022] In another example, the solvent system contains one or more components selected from the group consisting of a membrane permeability regulator, an enzyme activator, and a capillary dilator.

[0023] In one example, the solvent system contains ethanol, propylene carbonate, acetone, and phosphoric acid. In another example, the solvent system contains ethanol, propylene glycol, acetone, and phosphoric acid. In another example, the solvent system contains ethanol, propylene glycol, propylene carbonate, acetone, and phosphoric acid.

[0024] In another example, the solvent system contains ethanol, propylene carbonate, acetone, lemon oil, vitamin E, phytantriol, dexpanthenol, lauricidin, methylsulfonylmethane (MSM), forskolin, and phosphoric acid.

[0025] In another example, the amount of phosphoric acid is in the range of 0.2 to 2 in terms of the molecular ratio of phosphoric acid to insulin. In another example, the formulation contains 1.18 moles of phosphoric acid per mole of insulin.

[0026] In one example, the insulin contained in the formulation described herein has a molecular weight in the range of 340 daltons to 22,000 daltons.

[0027] In another example, the insulin is human insulin.

[0028] In one example, the molecular characteristics of insulin and the solvent system are substantially similar. The molecular characteristics can be van der Waals forces or dipole moments.

[0029] In another example, the formulation comprises insulin and a solvent system, where the solvent system comprises one or more selected from the group consisting of a solvent, a solvent modifying factor, a solute modifying factor, a source of cell activation energy, and a skin stabilizer, and optionally comprises one or more selected from the group consisting of a membrane permeability regulating factor, an enzyme activating factor, and a capillary dilating factor, and where insulin and the solvent system exhibit substantially similar van der Waals forces and / or dipole moments.

[0030] In one example, the formulation is a topical formulation.

[0031] In one example, the formulation is formulated in a liquid dosage form. In one example, the liquid dosage form can range from 0.1 mL to 1 mL. In another example, the liquid dosage form contains at least one type of insulin in the range of an insulin amount from 1 IU / mL to 1000 IU / mL.

[0032] In some examples, the dosage form of the transdermal insulin formulation disclosed herein includes a liquid dosage form, such as a solution, a liquid spray, a lotion, etc.

[0033] In some examples, the dosage form of the transdermal insulin formulation disclosed herein may be applied to any site of the skin, such as the forearm, upper arm, back, and chest.

[0034] In one example, the transdermal insulin formulation described herein is administered at a dosage in the range of an insulin amount from 1 IU / day to 1000 IU / day.

[0035] In some examples, the transdermal insulin formulation described herein may be designed as an immediate-release and transdermally absorbable insulin, or a sustained-release and transdermally absorbable insulin over a long period of time.

[0036] In one example, a method for delivering insulin to a subject in need thereof is disclosed herein, the method comprising administering to the subject a therapeutically effective amount of the formulation described herein.

[0037] In one example, a method for stabilizing glucose levels in a subject receiving insulin, comprising administering to the subject a therapeutically effective amount of the formulation described herein, is disclosed herein.

[0038] In one example, a method for treating diabetes, comprising administering to the subject a therapeutically effective amount of the formulation described herein, is disclosed herein.

[0039] In one example, a method for reducing hypoglycemia, comprising administering to the subject a therapeutically effective amount of the formulation described herein, is disclosed herein. In one example, a method for delivering insulin while minimizing hypoglycemia, comprising administering to the subject a therapeutically effective amount of the formulation described herein, is disclosed herein.

[0040] In one example, a method for rapidly delivering at least 90% of at least one type of insulin to the subcutaneous adipose tissue stroma and / or capillary network by skin penetration is disclosed herein. This delivery may be achieved in just a few tens of seconds or less than a few minutes.

[0041] In one example, a transdermal insulin formulation for treating a living body by rapidly delivering at least one type of insulin in an effective amount across the skin by applying the transdermal insulin formulation to an area of the skin is disclosed herein. The transdermal insulin formulation includes the at least one type of insulin and a solvent system. The at least one type of insulin has a molecular weight greater than 300 Daltons. The at least one type of insulin has molecular properties including van der Waals forces and dipole moments. The at least one type of insulin is dissolved as a solute in the solvent system. The solvent system has molecular properties including van der Waals forces and dipole moments. The molecular properties of the solvent system change the total dielectric constant of the solute + solvent system to be substantially the same or approximately ±20%.

[0042] The present application also discloses a method for preparing a transdermal insulin formulation as described herein. In one example, the method includes the following: (a) Selecting at least one type of insulin; (b) Determining an effective dose of the at least one type of insulin, wherein the effective dose of the at least one type of insulin has molecular properties including van der Waals forces and dipole moments; (c) Quantifying the molecular properties of the at least one type of insulin; (d) Determining an amount of a solvent system for solubilizing the effective dose of the at least one type of insulin, wherein the amount of the solvent system has molecular properties including van der Waals forces and dipole moments; (e) Quantifying the molecular properties of the amount of insulin; (f) Comparing the molecular properties of the at least one type of insulin with the molecular properties of the solvent system; (g) Determining that the molecular properties of the solvent system without solute are substantially the same as or approximately ±20% of the molecular properties of the at least one type of insulin; and (h) Combining the solvent system and the at least one type of insulin to provide a transdermal insulin formulation.

[0043] In one example, a method for preparing a transdermal insulin formulation as described herein includes selecting one or more components of a solvent system to determine the amount of the solvent system for solubilizing the effective dose of the at least one type of insulin, wherein the one or more components are selected from the group consisting of a solvent, a solvent modifying factor, a solute modifying factor, a source of cell activation energy, a skin stabilizer, and combinations thereof; each of the one or more components has different molecular properties including van der Waals forces and dipole moments.

[0044] In another example, a method for preparing a transdermal insulin formulation as described herein includes selecting one or more components of a solvent system to determine the amount of the solvent system for solubilizing the effective dose of the at least one type of insulin, wherein the one or more components are selected from the group consisting of a solvent, a solvent modifying factor, a solute modifying factor, a source of cell activation energy, a skin stabilizer, one type of membrane permeability regulator, at least one type of enzyme activation factor, at least one type of capillary dilator, and combinations thereof; each of the one or more components has similar molecular properties including van der Waals forces and dipole moments.

[0045] In one example, a method of selecting components and amounts for preparing a transdermal insulin formulation as described herein is disclosed herein, where the method includes the following steps: (a) Selecting at least one type of insulin required to treat a specific condition; (b) Quantifying the amount of the insulin for the effective dose; (c) Quantifying the molecular properties of the insulin to include van der Waals forces and dipole moments; (d) Investigating a solvent for the insulin; (e) Quantifying the amount of the solvent to solubilize the insulin; (f) Quantifying the molecular properties of the solvent to include van der Waals forces and dipole moments; (g) Comparing the molecular properties of the solvent with the molecular properties of the insulin; (h) Determining additional components to form a solvent system for the transition; (i) Quantifying the molecular properties of the additional components to include van der Waals forces and molar moments; (j) Determining the weighted sum of the molecular properties of the additional components and the molecular properties of the solvent to determine the molecular properties of the solvent system; (k) Summing the molecular properties of the solvent system + the insulin; (l) Comparing (j) and (k); and (m) Selecting the solvent system, wherein the molecular properties of the at least one type of insulin in the solvent system are substantially the same as the molecular properties of the solvent system without insulin.

[0046] In one example described herein, the van der Waals forces and / or dipole moments of at least one type of insulin in the solvent system are approximately ±20% of the van der Waals forces and / or dipole moments of the solvent system without insulin. In one example described herein, the van der Waals forces and / or dipole moments of at least one type of insulin in the solvent system are approximately ±15% of the van der Waals forces and / or dipole moments of the solvent system. In one example described herein, the van der Waals forces and / or dipole moments of at least one type of insulin in the solvent system are approximately ±10% of the van der Waals forces and / or dipole moments of the solvent system. In one example described herein, the van der Waals forces and / or dipole moments of at least one type of insulin in the solvent system are approximately ±5% of the van der Waals forces and / or dipole moments of the solvent system.

Brief Description of the Drawings

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Mode for Carrying Out the Invention

[0048] Definition With reference to the non-limiting examples specifically shown in the accompanying drawings and detailed in the following description, the examples described herein and their various features and advantages will be explained in more detail. Descriptions of well-known features and processing techniques are omitted so as not to unnecessarily obscure the embodiments described herein. The examples used herein are merely intended to facilitate the understanding of the methods by which the embodiments described herein can be implemented, and further, are intended to enable those skilled in the art to implement the embodiments described herein. Therefore, those examples should not be regarded as limiting the scope of the embodiments described herein.

[0049] The definitions and embodiments described in this section and in other sections are intended to apply to all embodiments and aspects of the present application described herein, as appropriate and understood by one of ordinary skill in the art, unless otherwise specifically indicated elsewhere.

[0050] As used herein, the singular forms "a", "an", and "the" include plural references unless the context clearly dictates otherwise. For example, an embodiment that includes "a species of agent" is to be understood to represent a particular aspect that includes one species of compound or two or more additional compounds.

[0051] The terms "about", "substantially", and "approximately" as used herein mean a reasonable degree of variation for the matter being modified such that the end result is not significantly changed. These terms of degree should be construed to include a variation of at least ±5%, at least ±10%, at least ±15%, or at least ±20% of the matter being modified if such variation would not negate the meaning of the modified term.

[0052] The term "suitable" as used herein means that the selection of a compound or condition will depend on the particular synthetic operation being performed and the nature of the molecule being transformed, but the selection will be well within the skill of one of ordinary skill in the art. All steps of the processes / methods described herein are performed under conditions sufficient to provide the described products. One of ordinary skill in the art will understand that, for example, all reaction conditions, including reaction solvent, reaction time, reaction temperature, reaction pressure, reactant ratio, and whether the reaction should be carried out in an anhydrous or inert atmosphere, can be varied to optimize the yield of the desired product and that this is within the skill of one of ordinary skill in the art.

[0053] The terms "agent" and "component" as used herein are synonymous and refer to a compound or mixture of compounds that, when added to a formulation, tends to produce a particular effect on the properties of that formulation.

[0054] The term "and / or" in this specification means that the listed items exist, or are used, individually or in combination. In fact, this term means that "at least one kind" or "one kind or more" of the listed items is used, or exists.

[0055] The term "delivery solution" in this specification refers to a liquid or semi-solid mixture of chemicals that can be broadly classified as solvents, solvent modifiers, and / or other chemical mediators in which the above-mentioned pharmaceutical active ingredient is stably dissolved in order to act as a vehicle for introducing the active pharmaceutical ingredient into the physiological environment, regardless of whether it is through injection, ingestion, or through the skin. In this context, the term "pharmaceutical active ingredient" refers to insulin and other compounds used to treat diabetes.

[0056] The term "delivery system" in this specification refers to the above-mentioned "delivery solution" formulated in specific ratios, ranges of ratios, and values relative to each other for a solution in which the above-mentioned pharmaceutical active ingredient is stably dissolved in order to act as a vehicle for introducing the active pharmaceutical ingredient into the physiological environment, regardless of whether it is through injection, ingestion, or through the skin.

[0057] The terms "formulation", "composition", "pharmaceutical formulation", and "pharmaceutical composition" in this specification are synonyms and refer to formulations for pharmaceutical use.

[0058] The term "pharmaceutically acceptable" in this specification refers to a material that does not cancel out the biological activity or properties of the agents described in this specification and is relatively non-toxic (i.e., the toxicity of the material is significantly outweighed by the advantages of the material). In some cases, a pharmaceutically acceptable material is administered to an individual without causing significant undesirable biological effects or interacting significantly in a harmful manner with any of the components of the formulation in which it is contained. The term "pharmaceutically acceptable" also refers to being suitable for the treatment of animals, such as humans.

[0059] As used herein, the term "effective amount" means an amount sufficient to achieve a desired result and thus varies depending on the component and its desired result. However, once the desired effect is clear, determining the effective amount is within the skill of the art.

[0060] As used herein and known in the art, the terms "treat" and "treatment" mean an approach for obtaining a benefit or desired result (including clinical results). A beneficial or desired clinical result can be detectable or undetectable and includes, but is not limited to, alleviation or improvement of one or more symptoms or conditions, attenuation of the disease state, stabilization of the disease condition (i.e., not getting worse), prevention of disease spread, delay or slowing of disease progression, improvement or alleviation of the disease state, reduction of disease recurrence, and remission (whether partial or complete). "Treat" and "treatment" can also mean extending survival compared to the expected survival without treatment. As used herein, "treat" and "treatment" include prophylactic measures. A method of treatment includes administering to a subject a therapeutically effective amount of a formulation as described herein and optionally consisting of a single administration or including a series of applications. The length of the treatment period depends on various factors such as the severity of the condition, the age of the patient, the concentration of the active ingredient or agent, the activity of the formulation described herein, and / or combinations thereof. It will also be understood that the effective dosage of a formulation used for treatment or prevention may increase or decrease during the course of a particular treatment or prevention regimen. Changes in dosage will be made or clarified by known standard diagnostic assays in the art. Sometimes, long-term administration may be required. For example, the formulation is administered to the subject in an amount sufficient to treat the subject and for a period of time.

[0061] As used herein, the term "topical formulation" includes formulations suitable for topical application to the skin. Topical formulations may be used, for example, to provide a therapeutic advantage to their user. Particular topical formulations may be used for local, site-specific, or transdermal application of a substance.

[0062] The term "transdermal" as used herein includes processes that occur through the skin. The terms "transdermal", "percutaneous", and "transcutaneous" may be used synonymously. In certain embodiments, "transdermal" also includes epicutaneous. Transdermal administration is often applied when systemic delivery of the active ingredient is desired, but it can also be useful for delivering the active ingredient to the underlying tissue of the skin with minimal systemic absorption.

[0063] The term "transdermal application" as used herein includes administration through the skin. Transdermal application can be used for systemic delivery of the active ingredient, but it can also be useful for delivering the active ingredient to the underlying tissue of the skin with minimal systemic absorption. In certain embodiments, "transdermal application" can also include application on the skin.

[0064] The term "pharmaceutically acceptable salt" means an acid addition salt or a base addition salt that is suitable or compatible for the treatment of subjects, including human subjects.

[0065] The term "diabetes" as used herein is intended to mean all diabetic conditions, including, but not limited to, diabetes, hereditary diabetes, type 1 diabetes, type 2 diabetes, type 3 diabetes, type 4 adult-onset, type 5 young-onset adult (diabetes) (MODY), and gestational diabetes. The term "diabetes" also refers to a chronic disease characterized by a relative or absolute lack of insulin that results in abnormal glucose tolerance. Type 1 diabetes is also known as insulin-dependent diabetes mellitus (IDDM) and includes, for example, juvenile-onset diabetes. Type 1 is primarily caused by the destruction of pancreatic beta cells. Type 2 diabetes, also known as non-insulin-dependent diabetes mellitus (NIDDM), is characterized in part by a reduced postprandial insulin release. Insulin resistance can also be a factor leading to the development of type 2 diabetes. Type 3 diabetes is caused by trauma to insulin-producing tissues, resulting in a cessation or significant reduction in insulin production. Type 4 diabetes is caused by insulin resistance in the elderly who are not overweight or obese. Type 5 diabetes or MODY5 or hereditary diabetes is a form of diabetes caused by a mutation in a single gene. This mutation causes abnormal pancreatic beta cell function, resulting in insufficient insulin production. In some cases, insulin resistance develops. Gestational diabetes occurs during pregnancy in response to hormonal changes that occur during pregnancy.

[0066] The term "diabetes" is also intended to include individuals with hyperglycemia, including chronic hyperglycemia, hyperinsulinemia, abnormal glucose homeostasis or glucose intolerance, and insulin resistance. Plasma glucose levels in hyperglycemic individuals include glucose concentrations higher than normal, as determined, for example, by reliable diagnostic indices. Such hyperglycemic individuals are at risk or predisposed to presenting clear clinical symptoms of diabetes.

[0067] Numerical ranges herein are intended to include all numbers and sub-numbers within the range, whether specifically disclosed or not. Further, these numerical ranges should be considered as providing a basis for claims directed to all or sub-numbers within the range.

[0068] It is understood that all components used in the formulations of the present invention must be non-toxic and safe for human use within the applicable and recommended dosage ranges. Also, all amounts, quantities, percentages (%) in the following description and the appended patent claims are by weight, unless otherwise specified.

[0069] Formulation Disclosed herein is a transdermal insulin formulation comprising at least one type of insulin and a solvent system. Insulin potency varies from lot to lot and is defined in units of IU (International Unit) and is typically 28 IU / mg.

[0070] At least one type of insulin can be selected from the group consisting of rapid-acting insulin, short-acting insulin, intermediate-acting insulin, long-acting insulin, and mixtures thereof. In another example, at least one type of insulin is human insulin. At least one type of insulin included in the formulations described herein can have a molecular weight in the range of 340 Daltons to 22,000 Daltons.

[0071] In some examples, the formulation contains 10 IU / ml of insulin. In other examples, the formulation contains 50 IU / ml of insulin. In yet another example, the formulation contains 100 IU / ml of insulin. In yet another example, the formulation contains 200 IU / ml of insulin. In yet another example, the formulation contains 500 IU / ml of insulin. Thus, the insulin potency of the formulations described herein can be, for example, 10 IU / ml, 50 IU / ml, 100 IU / ml, 200 IU / ml, or 500 IU / ml.

[0072] In some examples, the formulations described herein are designed to be delivered in a predetermined amount. In some examples, the delivery system delivers 0.2 mL to 1 mL and contains an amount of insulin in the range of 7 to 1,700 IU / mL. In some examples, the formulation is prepared in a unit dosage form, where the volume of the unit dosage form ranges from 0.2 mL to 1 mL, and where the unit dosage form contains insulin in an amount ranging from 0.25 mg to 60 mg. In some examples, the volume of the unit dosage form is 0.2 mL, 0.3 mL, 0.4 mL, 0.5 mL, 0.6 mL, 0.7 mL, 0.8 mL, 0.9 mL, or 1.0 mL. In some examples, the unit dosage form contains insulin in an amount of 7 IU, 14 IU, 28 IU, 140 IU, 280 IU, 350 IU, 420 IU, 490 IU, 560 IU, 630 IU, 700 IU, 770 IU, 840 IU, 910 IU, 980 IU, 1,050 IU, 1,120 IU, 1,190 IU, 1,260 IU, 1,330 IU, 1,400 IU, 1,470 IU, 1,540 IU, 1,610 IU, or 1,680 IU. In some examples, the unit dosage form contains insulin in an amount within the range of amounts described in this paragraph.

[0073] In one example, a transdermal insulin formulation as described herein is administered within a dosage range of 1 IU / dose to 750 IU / dose of insulin, 40 IU / dose to 120 IU / dose of insulin, 0.5 IU / dose to about 5 IU / dose of insulin; and 50 IU / dose to 500 IU / dose of insulin.

[0074] In some examples, a transdermal insulin formulation as described herein can be designed to deliver as immediate-release insulin. In other embodiments, a transdermal insulin formulation as described herein can be designed as a sustained-release insulin that is effective over a long period of time by transdermal absorption.

[0075] Rapid-acting insulin fills the insulin deficit for meals consumed simultaneously with the injection. Short-acting insulin fills the insulin deficit for meals consumed within 30 to 60 minutes. Intermediate-acting insulin fills the insulin deficit for approximately half a day or overnight. This type of insulin is often combined with rapid-acting or short-acting insulin. Long-acting insulin fills the insulin deficit for almost the entire day. This type of insulin is often combined with rapid-acting or short-acting insulin as needed. Representative insulins are shown in Table 1 below.

[0076]

Table 1

[0077] At least one type of insulin can be selected from the group consisting of rapid-acting insulin, short-acting insulin, intermediate-acting insulin, long-acting insulin, and mixtures thereof; and can be present in an amount in the range of 0.1% to 25% (wt / wt) of the total formulation amount.

[0078] In one example, a transdermal insulin preparation as described herein contains at least one type of insulin in an amount in the range of 0.1% to 25% (wt / wt) of the total formulation amount. In another example, at least one type of insulin is present in an amount in the range of 0.1% to 20% (wt / wt) of the total formulation amount. In yet another example, at least one type of insulin is present in an amount in the range of 0.1% to 15% (wt / wt) of the total formulation amount. In yet another example, at least one type of insulin is present in an amount in the range of 0.1% to 10% (wt / wt) of the total formulation amount. In yet another example, at least one type of insulin is present in an amount in the range of 0.1% to 7.5% (wt / wt) of the total formulation amount. In yet another example, at least one type of insulin is present in an amount in the range of 0.1% to 5% (wt / wt) of the total formulation amount. In yet another example, at least one type of insulin is present in an amount in the range of 0.1% to 2.5% (wt / wt) of the total formulation amount. In yet another example, at least one type of insulin is present in an amount in the range of 0.1% to 1% (wt / wt) of the total formulation amount. In yet another example, at least one type of insulin is present in an amount in the range of 0.1% to 0.5% (wt / wt) of the total formulation amount. In yet another example, at least one type of insulin is present in an amount in the range of 0.1% to 0.45% (wt / wt) of the total formulation amount. In yet another example, at least one type of insulin is present in an amount in the range of 0.1% to 0.40% (wt / wt) of the total formulation amount. In yet another example, at least one type of insulin is present in an amount in the range of 0.1% to 0.35% (wt / wt) of the total formulation amount. In yet another example, at least one type of insulin is present in an amount in the range of 0.1% to 0.30% (wt / wt) of the total formulation amount. In yet another example, at least one type of insulin is present in an amount in the range of 0.1% to 0.25% (wt / wt) of the total formulation amount. In yet another example, at least one type of insulin is present in an amount in the range of 0.1% to 0.20% (wt / wt) of the total formulation amount.

[0079] In one example, a transdermal insulin formulation as described herein contains at least one type of insulin in an amount of 0.1% (wt / wt) of the total formulation amount. In another example, at least one type of insulin is present in an amount of 0.15% (wt / wt) of the total formulation amount. In yet another example, at least one type of insulin is present in an amount of 0.20% (wt / wt) of the total formulation amount. In yet another example, at least one type of insulin is present in an amount of 0.3% (wt / wt) of the total formulation amount. In yet another example, at least one type of insulin is present in an amount of 0.4% (wt / wt) of the total formulation amount. In yet another example, at least one type of insulin is present in an amount of 0.5% (wt / wt) of the total formulation amount. In yet another example, at least one type of insulin is present in an amount of 0.6% (wt / wt) of the total formulation amount. In yet another example, at least one type of insulin is present in an amount of 0.7% (wt / wt) of the total formulation amount. In yet another example, at least one type of insulin is present in an amount of 0.8% (wt / wt) of the total formulation amount. In yet another example, at least one type of insulin is present in an amount of 0.9% (wt / wt) of the total formulation amount. In yet another example, at least one type of insulin is present in an amount of 1% (wt / wt) of the total formulation amount. In yet another example, at least one type of insulin is present in an amount of 1.5% (wt / wt) of the total formulation amount. In yet another example, at least one type of insulin is present in an amount of 2% (wt / wt) of the total formulation amount. In yet another example, at least one type of insulin is present in an amount of 2.5% (wt / wt) of the total formulation amount. In yet another example, at least one type of insulin is present in an amount of 5% (wt / wt) of the total formulation amount. In yet another example, at least one type of insulin is present in an amount of 7.5% (wt / wt) of the total formulation amount. In yet another example, at least one type of insulin is present in an amount of 10% (wt / wt) of the total formulation amount. In another example, at least one type of insulin is present in an amount of 12.5% (wt / wt) of the total formulation amount.In yet another example, at least one type of insulin is present in an amount of 15% (wt / wt) of the total formulation amount. In yet another example, at least one type of insulin is present in an amount of 17.5% (wt / wt) of the total formulation amount. In yet another example, at least one type of insulin is present in an amount of 20% (wt / wt) of the total formulation amount. In yet another example, at least one type of insulin is present in an amount of 22.5% (wt / wt) of the total formulation amount. In yet another example, at least one type of insulin is present in an amount of 25% (wt / wt) of the total formulation amount. In yet another example, at least one type of insulin is present in an amount within the range mentioned in this paragraph.

[0080] Solvent system Disclosed herein is a transdermal insulin formulation comprising at least one type of insulin and a solvent system, wherein the solvent system comprises one or more components selected from the group consisting of two or more solvents, at least one solvent modifier, at least one solute modifier, at least one source of cell activation energy, and at least one skin stabilizer.

[0081] Solvent The solvent is the main component of the carrier of insulin and is preferably one in which insulin is soluble or at least substantially soluble, or can be made soluble or more soluble by the addition of one or more solvent modifiers. The expression "substantially soluble" herein means that the minimum effective dose of insulin, generally at least 0.25 mg, preferably at least 0.5 mg, ideally at least 1 mg, dissolves in 1 mL of the solvent or in 1 mL of a mixture of solvents containing a solvent modifier.

[0082] Preferred solvents include lower alcohols having about 2 to about 6 carbon atoms, preferably 2 to 4 carbon atoms, such as, for example, monoalcohols such as ethanol, isopropanol, sec-butanol, etc., or, for example, polyols such as ethylene glycol, propylene glycol, propylene carbonate, butylene glycol, glycerol, etc. A mixture of solvents may be used. Other solvents such as ketones (e.g., acetone, methyl ethyl ketone), ethers (e.g., ethyl ether), etc. may also be used in amounts that are safe and non-toxic in use.

[0083] The solvent system is generally non-aqueous, but water may also be added as one element of the other components, such as, for example, an azeotropic mixture of alcohol and water. When water is present in the solvent, it usually constitutes less than about 50%, preferably less than about 10%, particularly preferably less than about 2% by weight of the total solvent, but more or less may be used. Further, as will be apparent from the examples described hereinafter, formulations using the principles disclosed in the present application and further described in more detail hereinbelow may also be formulated as aqueous emulsions, including the case where the aqueous phase is the main phase and the continuous phase. Such aqueous emulsions are rapidly absorbed in less than 1 minute and release insulin, as in the case of a non-aqueous (usually less than about 5%, particularly less than about 2% water) solvent system.

[0084] The total amount of the solvent can be selected to ensure the dissolution of insulin and other additives and to provide a suitable product viscosity. Based on the total weight of the formulation, an amount of solvent in the range of 5% wt / wt to 90% wt / wt, preferably 25% wt / wt to 75% wt / wt can be used.

[0085] Solvent modification factor Solvent modifying factors for insulin delivery systems, such as those presented herein, are selected to adjust the polarity of the solvent. The solvent modifying factor or mixture of solvent modifying factors enables the solvent system [comprising the solvent and the solvent modifying factor] to form a weak complex with insulin (i.e., a bond by van der Waals forces), thus resulting in a stable formulation having a high insulin: solvent ratio. "Stable" herein is intended to have its ordinary meaning, i.e., that the formulation can be stored at room temperature or higher temperatures for at least one day, usually for at least 30 days, without undergoing phase separation. A "high insulin: solvent" ratio means at least 50 IU of insulin per mL of solvent (or solvent + modifier), and more generally means an amount of insulin that exceeds the solubility of insulin in the solvent alone or in each solvent of a multi - solvent system.

[0086] One or more of lemon oil (or / and d - limonene), vitamin E, provitamin B, D - panthenol, and methylsulfonylmethane (MSM) can be used as solvent modifying factors in the transdermal insulin formulations described herein.

[0087] The amount of the solvent modifying factor can be selected to yield the desired insulin: solvent ratio and depends on various factors, for example, mainly the polarity of each component (including the solvent, the solvent modifying factor, and insulin), and factors including polarizability, dipole moment, and van der Waals forces.

[0088] In this regard, to balance the polarity and dipole moment of insulin with those of the solvent system, the amounts of the individual components of the solvent system can be selected such that the weighted (molar) average of the dipole moments of the individual components is substantially the same as the dipole moment of the solution containing dissolved insulin in the base system (empty system).

[0089] Desired Insulin: The amount of the solvent modification factor suitable for obtaining the solvent ratio can be in the range of 0.0001% wt / wt to 50% wt / wt, preferably 0.1% wt / wt to 35% wt / wt, more preferably 0.1% wt / wt to 5% wt / wt, based on the total weight of the formulation.

[0090] Solute modification factor The solute modification factor may be included in the formulation of the transdermal insulin preparation to facilitate the dissolution of insoluble or slightly soluble insulin at a higher concentration. A solute modifier that forms a reversible or transient complex with insulin to promote passage through the skin while minimizing the immune response is particularly effective. The solute modification factor may also preferably be a nutrient compound that can be metabolized by the body when insulin is released from the complex.

[0091] Examples of solute modification factors include terpenes, oxindole alkaloids, quercitrin (a glycoside of quercetin), genistein and its glucosides, genistein, polyphenolic flavonoids, and other sugar adduct glucuronides (such as scutellarin, trans-ferulic acid, alpha-lipoic acid, etc.), sterols (such as cholesterol and cholesterol-like compounds, etc.), and hormones (isoflavones, 3,3'-thiodipropionic acid (sulfone-oxidized propionic acid), phosphatidylserine and choline, vitamin D3, vitamin K1, dehydroepiandrosterone (DHEA), etc.). Further suitable candidate compounds include, for example, berberine, black pepper (such as Bioperine®), phosphatidylserine, phosphatidylcholine. Another group of candidate compounds includes boswellic acid, Hypericum (St. John's wort), and phytic acid.

[0092] The selection of a specific solute modification factor promotes the movement of the insulin-complex through the stratum corneum and viable skin into its optimal targeted internal circulation system in the interstitial, blood, or lymphatic fluid.

[0093] The suitable amount of the solute modifier can be determined based on factors such as, for example, the solubility of the modifier in the system (e.g., solvent + solvent modifier), the compatibility of its molecule with insulin, and the ability to regulate the polarization rate of insulin that increases the concentration (solubility) of insulin in the solvent. The amount of the solute modifier can be in the range of 0.003% to 5%, preferably 0.1% to 5%, more preferably 0.1% to 4% of the total weight of the formulation. The amount of the solute modifier or solute modifying agent may be equivalent to the amount of insulin such that it provides a 1:1 interaction between the modifier and insulin.

[0094] The above-mentioned modifiers, i.e., the solvent modifier and the solute modifier, and other components of the solvent / carrier delivery system can be selected from substances that the body recognizes as available components of other physiological systems. This selection thus promotes the almost complete dissociation of insulin from the delivery system when it enters the body. Since these carrier / complex compounds are reducible to physiological basic components, they are not harmful to the body.

[0095] Source of cell activation energy The transdermal insulin formulation described herein includes a source of cell activation energy, which serves to induce a high concentration of enzyme-substrate complex formed, for example, by the activation of the N (stimulatory) protein of adenylate cyclase, thereby bringing about the cellular level of adenosine 3′,5′-cyclic monophosphate (cAMP) approaching the upper limit of the intracellular cAMP concentration.

[0096] Examples of such agents include extracts of the plant Coleus Forskohlii, and in particular, forskolin, which is a labdane diterpenoid. Other extracts of Coleus Forskohlii, such as colforsin or coleonol, can also be used.

[0097] As other examples of activation energy sources for promoting the production of cAMP via either a precursor or a cell activator, for example, methylxanthines, psychogenin and saponins, Angelicae dahuricae radix (producing angelic acid), feropterin, oxyposidanin may be mentioned.

[0098] Substances that stimulate the intracellular production of cGMP can also be used, and they may be selected from the group consisting of acetylcholine, cytidine diphosphocholine, and ascorbic acid (vitamin C).

[0099] The amount of the source of cell activation energy depends on factors such as, for example, the activation (positive or negative) energy when insulin encounters its receptor (increasing or decreasing cAMP or cGMP levels) for the insulin mechanism of action. The preferred amount of forskolin or acetylcholine or other sources of cell activation energy can be in the range of 0.001% to 0.1%, preferably 0.001% to 0.01%, more preferably 0.001% to 0.005% based on the total weight of the formulation.

[0100] Skin stabilizer A skin stabilizer may be included in the transdermal insulin formulation described herein to stabilize the skin before passage and to help the skin repair any damage resulting from the transfer of insulin and solvents and other components in the formulation.

[0101] Function as a skin stabilizer, and examples of substances that may be included in the formulations described herein include glycerol monolaurate (e.g., lauricidin (registered trademark)) and similar fatty acid esters, vitamin D3, alkoxyglycerols, unsaturated fatty acids (eicosapentaenoic acid (EPA), docosahexaenoic acid (DHA), and gamma-linolenic acid (GLA), vitamin E (alpha tocopherol acetate), etc.), esters (e.g., acetic acid and its derivatives (e.g., tocotrienols, D-panthenol, phytantriol, dehydroepiandrosterone (DHEA), pregnenolone, pregnenolone acetate, escrin, allantoin, ascorbyl palmitate), etc.).

[0102] A suitable amount of the skin stabilizer can be determined based on factors such as the type of reaction, e.g., between insulin and the skin, between the solvent and the skin, etc. The amount of the skin stabilizer, if present, can be 0.01% in an example of the formulations described herein. Further, the skin stabilizer can be present in the range of 0.05% to 5%, preferably 0.1% to 5%, more preferably 0.1% to 2% by weight based on the total formulation. It is preferred to select a stabilizer that is effective in stabilizing the skin at the lowest possible concentration.

[0103] Other components Enzyme activation factor / signal transduction compound In one example, a transdermal insulin formulation as described herein may include enzyme activation factors / signal transduction compounds such as forskolin and sulforaphane.

[0104] A suitable amount of such enzyme activation factor / signal transduction compound can be in the range of 0.01% to 0.05%, preferably 0.01% to 0.02% by weight based on the total formulation.

[0105] Disclosed herein is a transdermal insulin formulation comprising at least one type of insulin and a solvent system.

[0106] In one example disclosed herein, the transdermal insulin formulation comprises at least one type of insulin and a solvent system, where the solvent system without insulin comprises molecular properties substantially similar to the molecular properties of the solute in the system. In another example, the transdermal insulin formulation comprises at least one type of insulin and a solvent system, where the solvent system without insulin comprises molecular properties corresponding to approximately ±20% of the molecular properties of the solute in the solution. In another example, the transdermal insulin formulation comprises at least one type of insulin and a solvent system, where the solvent system without insulin comprises molecular properties corresponding to approximately ±15% of the molecular properties of the solute in the solution. In another example, the transdermal insulin formulation comprises at least one type of insulin and a solvent system, where the solvent system without insulin comprises molecular properties corresponding to approximately ±10% of the molecular properties of the solute in the solution. In another example, the transdermal insulin formulation comprises at least one type of insulin and a solvent system, where the solvent system without insulin comprises molecular properties corresponding to approximately ±5% of the molecular properties of the solute in the solution.

[0107] The molecular properties can be selected from van der Waals forces and / or dipole moment.

[0108] In this regard, it is understood that the dipole moment of a given compound may be obtained directly from the literature if available, or may be measured or calculated by standard techniques including commercially available chemical modeling software packages. Generally, the dipole moment is determined experimentally for an element or compound by suspending the molecule in an electromagnetic field and measuring the amount of energy (torque) required to rotate the molecule once. The dipole moment correlates with van der Waals forces, the number of hydrogen bonds, and the electrostatic energy of the molecule. Two chemical substances having approximately the same dipole moment usually have an affinity for each other and attract each other without the need for a covalent bond.

[0109] To determine the dipole moments of the solvent and the modifier, a weighted average of the dipole moments of the individual components is used. The weighted average should be fairly close to the dipole moment of the solute. The closer the match, the faster the rate of movement through the skin. The delivery system, if necessary, moves the dipole moment of the system containing the modifier and other additives, including the solute, as close as possible, preferably within 15%, particularly within 10%, and even more particularly within 5% of the dipole moment of the solute, to the dipole moment of the insulin-free delivery system.

[0110] More specifically, in accordance with the preferred method for preparing the formulations described herein, particularly for increasing the amount of insulin that can be stably transported in solution in the transdermal delivery formulations described herein, the selection of the components of the solvent system and other functional additives and the amounts thereof may first be determined by balancing the dipole moment of insulin against the dipole moment of the final formulation. The dipole moment of the final formulation is selected to be the weighted average dipole moment of each of the individual components. The weighted average is obtained by calculating the sum of the dipole moments of the components, where the dipole moment is obtained by multiplying the dipole moment of the component by the molar amount of the component in a given volume, for example, 100 cc. For this calculation, each component in the formulation is assumed to act independently of the other components. Thus, for example, the dipole moment of any particular component does not take into account the electronic effects of the other components, such as repulsive or attractive effects. However, by considering the concentration, that is, by multiplying the individual dipole moments by the molar concentration, a reasonable approximation of the match of the system properties for balancing with insulin is achieved.

[0111] As described herein, similar to the equilibration of dipole moments, formulations of solvent systems for insulin, when insulin is added to the solvent system, as a solubility predictor of a desired amount of insulin, the total molar - van der Waals forces of the solvent system containing insulin are within ±20%, preferably within ±15%, particularly preferably within ±10%, and most preferably within ±5% of the total molar - van der Waals forces of the solvent system without insulin, whereby the molar - van der Waals forces may be equilibrated.

[0112] When the difference between the total molar - van der Waals forces of the solvent system + insulin exceeds about 20%, particularly more than 15% of the total molar - van der Waals forces of the solvent system without insulin, the desired amount of insulin may tend to be insoluble in the solvent system or may precipitate from the solution upon standing overnight.

[0113] The transdermal insulin formulations as described herein provide delivery of at least about 90% or more of at least one type of insulin rapidly through the skin to the underlying adipose tissue. This delivery can be achieved in just a few tens of seconds or less than a few minutes.

[0114] The solvent system contains one or more components selected from the group consisting of at least two types of solvents, at least one type of solvent modifier, at least one type of solute modifier, at least one source of cell activation energy, at least one skin stabilizer, at least one membrane permeability regulator, at least one enzyme activator, and at least one capillary dilator.

[0115] In a preferred example, the solvent system contains one or more components selected from the group consisting of at least two types of solvents, at least one type of solvent modifier, at least one type of solute modifier, at least one source of cell activation energy, and at least one skin stabilizer.

[0116] In a more preferred example, the solvent system includes at least one component selected from the group consisting of at least two solvents, at least one solvent modifying factor, at least one source of cell activation energy, and at least one skin stabilizer.

[0117] In one example, the present specification discloses a transdermal preparation containing at least one insulin and a solvent system, where the solvent system includes two solvents, a solvent modifying factor, a source of cell activation energy, and a skin stabilizer.

[0118] In one example, the solvent may include at least one component selected from the group consisting of ethanol, isopropanol, ethylene glycol, propylene carbonate, propylene glycol, acetone, and methyl ethyl ketone. In a preferred example, the solvent includes at least one component selected from the group consisting of ethanol, propylene carbonate, propylene glycol, and acetone. In a more preferred embodiment, the solvent includes ethanol, propylene carbonate, and acetone.

[0119] In one example, the solvent system contains ethanol in an amount of 35% (wt / wt) of the total formulation amount. In another example, the solvent system contains ethanol in an amount of 36% (wt / wt) of the total formulation amount. In yet another example, the solvent system contains ethanol in an amount of 37% (wt / wt) of the total formulation amount. In yet another example, the solvent system contains ethanol in an amount of 38% (wt / wt) of the total formulation amount. In yet another example, the solvent system contains ethanol in an amount of 39% (wt / wt) of the total formulation amount. In yet another example, the solvent system contains ethanol in an amount of 40% (wt / wt) of the total formulation amount. In yet another example, the solvent system contains ethanol in an amount of 41% (wt / wt) of the total formulation amount. In yet another example, the solvent system contains ethanol in an amount of 42% (wt / wt) of the total formulation amount. In yet another example, the solvent system contains ethanol in an amount of 43% (wt / wt) of the total formulation amount. In yet another example, the solvent system contains ethanol in an amount of 44% (wt / wt) of the total formulation amount. In yet another example, the solvent system contains ethanol in an amount of 45% (wt / wt) of the total formulation amount. In yet another example, the solvent system contains ethanol in an amount of 46% (wt / wt) of the total formulation amount. In yet another example, the solvent system contains ethanol in an amount of 47% (wt / wt) of the total formulation amount. In yet another example, the solvent system contains ethanol in an amount of 48% (wt / wt) of the total formulation amount. In yet another example, the solvent system contains ethanol in an amount of 49% (wt / wt) of the total formulation amount. In yet another example, the solvent system contains ethanol in an amount of 50% (wt / wt) of the total formulation amount. In yet another example, the solvent system contains ethanol in an amount within the range of amounts described in this paragraph.

[0120] In one example, the solvent system contains propylene carbonate in an amount of 40% (wt / wt) of the total formulation amount. In another example, the solvent system contains propylene carbonate in an amount of 41% (wt / wt) of the total formulation amount. In yet another example, the solvent system contains propylene carbonate in an amount of 42% (wt / wt) of the total formulation amount. In yet another example, the solvent system contains propylene carbonate in an amount of 43% (wt / wt) of the total formulation amount. In yet another example, the solvent system contains propylene carbonate in an amount of 44% (wt / wt) of the total formulation amount and propylene carbonate. In yet another example, the solvent system contains propylene carbonate in an amount of 45% (wt / wt) of the total formulation amount. In yet another example, the solvent system contains propylene carbonate in an amount of 46% (wt / wt) of the total formulation amount. In yet another example, the solvent system contains propylene carbonate in an amount of 47% (wt / wt) of the total formulation amount. In yet another example, the solvent system contains propylene carbonate in an amount of 48% (wt / wt) of the total formulation amount. In yet another example, the solvent system contains propylene carbonate in an amount of 49% (wt / wt) of the total formulation amount. In yet another example, the solvent system contains propylene carbonate in an amount of 50% (wt / wt) of the total formulation amount. In yet another example, the solvent system contains propylene carbonate in an amount of 51% (wt / wt) of the total formulation amount. In yet another example, the solvent system contains propylene carbonate in an amount of 52% (wt / wt) of the total formulation amount. In yet another example, the solvent system contains propylene carbonate in an amount of 53% (wt / wt) of the total formulation amount. In yet another example, the solvent system contains propylene carbonate in an amount of 54% (wt / wt) of the total formulation amount. In yet another example, the solvent system contains propylene carbonate in an amount of 55% (wt / wt) of the total formulation amount. In yet another example, the solvent system contains propylene carbonate in an amount within the range of amounts described in this paragraph.

[0121] In one example, the solvent system contains propylene glycol in an amount of 40% (wt / wt) of the total formulation amount. In another example, the solvent system contains propylene glycol in an amount of 41% (wt / wt) of the total formulation amount. In yet another example, the solvent system contains propylene glycol in an amount of 42% (wt / wt) of the total formulation amount. In yet another example, the solvent system contains propylene glycol in an amount of 43% (wt / wt) of the total formulation amount. In yet another example, the solvent system contains propylene glycol in an amount of 44% (wt / wt) of the total formulation amount. In yet another example, the solvent system contains propylene glycol in an amount of 45% (wt / wt) of the total formulation amount. In yet another example, the solvent system contains propylene glycol in an amount of 46% (wt / wt) of the total formulation amount. In yet another example, the solvent system contains propylene glycol in an amount of 47% (wt / wt) of the total formulation amount. In yet another example, the solvent system contains propylene glycol in an amount of 48% (wt / wt) of the total formulation amount. In yet another example, the solvent system contains propylene glycol in an amount of 49% (wt / wt) of the total formulation amount. In yet another example, the solvent system contains propylene glycol in an amount of 50% (wt / wt) of the total formulation amount. In yet another example, the solvent system contains propylene glycol in an amount of 51% (wt / wt) of the total formulation amount. In yet another example, the solvent system contains propylene glycol in an amount of 52% (wt / wt) of the total formulation amount. In yet another example, the solvent system contains propylene glycol in an amount of 53% (wt / wt) of the total formulation amount. In yet another example, the solvent system contains propylene glycol in an amount of 54% (wt / wt) of the total formulation amount. In yet another example, the solvent system contains propylene glycol in an amount of 55% (wt / wt) of the total formulation amount. In yet another example, the solvent system contains propylene glycol in an amount within the range of amounts described in this paragraph.

[0122] In one example, the solvent system contains acetone in an amount of 0.5% (wt / wt) of the total formulation amount. In another example, the solvent system contains acetone in an amount of 0.75% (wt / wt) of the total formulation amount. In yet another example, the solvent system contains acetone in an amount of 1.0% (wt / wt) of the total formulation amount. In yet another example, the solvent system contains acetone in an amount of 1.25% (wt / wt) of the total formulation amount. In yet another example, the solvent system contains acetone in an amount of 1.5% (wt / wt) of the total formulation amount. In yet another example, the solvent system contains acetone in an amount of 1.75% (wt / wt) of the total formulation amount. In yet another example, the solvent system contains acetone in an amount of 2.0% (wt / wt) of the total formulation amount. In yet another example, the solvent system contains acetone in an amount of 2.25% (wt / wt) of the total formulation amount. In yet another example, the solvent system contains acetone in an amount of 2.5% (wt / wt) of the total formulation amount. In yet another example, the solvent system contains acetone in an amount of 2.75% (wt / wt) of the total formulation amount. In yet another example, the solvent system contains acetone in an amount of 3.0% (wt / wt) of the total formulation amount. In yet another example, the solvent system contains acetone in an amount of 3.25% (wt / wt) of the total formulation amount. In yet another example, the solvent system contains acetone in an amount of 3.5% (wt / wt) of the total formulation amount. In yet another example, the solvent system contains acetone in an amount of 3.75% (wt / wt) of the total formulation amount. In yet another example, the solvent system contains acetone in an amount of 4.0% (wt / wt) of the total formulation amount. In yet another example, the solvent system contains acetone in an amount of 4.25% (wt / wt) of the total formulation amount. In yet another example, the solvent system contains acetone in an amount of 4.5% (wt / wt) of the total formulation amount. In yet another example, the solvent system contains acetone in an amount of 4.75% (wt / wt) of the total formulation amount. In yet another example, the solvent system contains acetone in an amount of 5.0% (wt / wt) of the total formulation amount. In yet another example, the solvent system contains acetone in an amount within the range of amounts described in this paragraph.

[0123] In some examples, the solvent system includes an acid selected from the group consisting of an Arrhenius acid, a mineral acid, an organic acid, a Bronsted-Lowry acid, a strong acid, a weak acid, a diprotic acid, and a triprotic acid. Examples of such acids include, but are not limited to, hydrochloric acid, phosphoric acid, perchloric acid, sulfuric acid, nitric acid, hydroiodic acid, lactic acid, oxalic acid, succinic acid, hydrobromic acid, nitrous acid, and ammonium ion, fluorosulfuric acid, trifluoromethanesulfonic acid, fluoroantimonic acid, formic acid, sulfurous acid, benzoic acid, carbonic acid, citric acid, and arsenic acid. The solvent system described herein may contain the acid in an amount in the range of 0.25% (wt / wt) to 3.0% (wt / wt).

[0124] In one example, the solvent system contains phosphoric acid in an amount of 0.25% (wt / wt) of the total formulation amount. In another example, the solvent system contains phosphoric acid in an amount of 0.5% (wt / wt) of the total formulation amount. In yet another example, the solvent system contains phosphoric acid in an amount of 0.75% (wt / wt) of the total formulation amount. In yet another example, the solvent system contains phosphoric acid in an amount of 1.0% (wt / wt) of the total formulation amount. In yet another example, the solvent system contains phosphoric acid in an amount of 1.25% (wt / wt) of the total formulation amount. In yet another example, the solvent system contains phosphoric acid in an amount of 1.5% (wt / wt) of the total formulation amount. In yet another example, the solvent system contains phosphoric acid in an amount of 1.75% (wt / wt) of the total formulation amount. In yet another example, the solvent system contains phosphoric acid in an amount of 2.0% (wt / wt) of the total formulation amount. In yet another example, the solvent system contains phosphoric acid in an amount of 2.25% (wt / wt) of the total formulation amount. In yet another example, the solvent system contains phosphoric acid in an amount of 2.5% (wt / wt) of the total formulation amount. In yet another example, the solvent system contains phosphoric acid in an amount of 2.75% (wt / wt) of the total formulation amount. In yet another example, the solvent system contains phosphoric acid in an amount of 3.0% (wt / wt) of the total formulation amount. In yet another example, the solvent system contains phosphoric acid in an amount within the range of amounts described in this paragraph.

[0125] In one example, the solvent system contains phosphoric acid, where the molar ratio of phosphoric acid to insulin is 0.2. In another example, the solvent system contains phosphoric acid, where the molar ratio of phosphoric acid to insulin is 0.25. In another example, the solvent system contains phosphoric acid, where the molar ratio of phosphoric acid to insulin is 0.3. In another example, the solvent system contains phosphoric acid, where the molar ratio of phosphoric acid to insulin is 0.35. In yet another example, the solvent system contains phosphoric acid, where the molar ratio of phosphoric acid to insulin is 0.4. In another example, the solvent system contains phosphoric acid, where the molar ratio of phosphoric acid to insulin is 0.45. In yet another example, the solvent system contains phosphoric acid, where the molar ratio of phosphoric acid to insulin is 0.5. In yet another example, the solvent system contains phosphoric acid, where the molar ratio of phosphoric acid to insulin is 0.6. In yet another example, the solvent system contains phosphoric acid, where the molar ratio of phosphoric acid to insulin is 0.7. In yet another example, the solvent system contains phosphoric acid, where the molar ratio of phosphoric acid to insulin is 0.8. In yet another example, the solvent system contains phosphoric acid, where the molar ratio of phosphoric acid to insulin is 0.9. In yet another example, the solvent system contains phosphoric acid, where the molar ratio of phosphoric acid to insulin is 1.0. In yet another example, the solvent system contains phosphoric acid, where the molar ratio of phosphoric acid to insulin is 1.1. In another example, the solvent system contains phosphoric acid, where the molar ratio of phosphoric acid to insulin is 0.15. In yet another example, the solvent system contains phosphoric acid, where the molar ratio of phosphoric acid to insulin is 1.2. In another example, the solvent system contains phosphoric acid, where the molar ratio of phosphoric acid to insulin is 1.25. In yet another example, the solvent system contains phosphoric acid, where the molar ratio of phosphoric acid to insulin is 1.3. In another example, the solvent system contains phosphoric acid, where the molar ratio of phosphoric acid to insulin is 1.35. In yet another example, the solvent system contains phosphoric acid, where the molar ratio of phosphoric acid to insulin is 1.4.In another example, the solvent system contains phosphoric acid, where the molar ratio of phosphoric acid to insulin is 1.45. In yet another example, the solvent system contains phosphoric acid, where the molar ratio of phosphoric acid to insulin is 1.5. In another example, the solvent system contains phosphoric acid, where the molar ratio of phosphoric acid to insulin is 1.55. In yet another example, the solvent system contains phosphoric acid, where the molar ratio of phosphoric acid to insulin is 1.6. In another example, the solvent system contains phosphoric acid, where the molar ratio of phosphoric acid to insulin is 1.65. In yet another example, the solvent system contains phosphoric acid, where the molar ratio of phosphoric acid to insulin is 1.7. In another example, the solvent system contains phosphoric acid, where the molar ratio of phosphoric acid to insulin is 1.75. In yet another example, the solvent system contains phosphoric acid, where the molar ratio of phosphoric acid to insulin is 1.8. In another example, the solvent system contains phosphoric acid, where the molar ratio of phosphoric acid to insulin is 1.85. In yet another example, the solvent system contains phosphoric acid, where the molar ratio of phosphoric acid to insulin is 1.9. In another example, the solvent system contains phosphoric acid, where the molar ratio of phosphoric acid to insulin is 1.95. In yet another example, the solvent system contains phosphoric acid, where the molar ratio of phosphoric acid to insulin is 2.0. In yet another example, the solvent system contains phosphoric acid, where the molar ratio of phosphoric acid to insulin is within the range of the ratios described in this paragraph.

[0126] In one example, the solvent system contains a solvent modifying factor in an amount of 0.001% (wt / wt) of the total formulation amount. In another example, the solvent system contains a solvent modifying factor in an amount of 0.0025% (wt / wt) of the total formulation amount. In yet another example, the solvent system contains a solvent modifying factor in an amount of 0.005% (wt / wt) of the total formulation amount. In yet another example, the solvent system contains a solvent modifying factor in an amount of 0.01% (wt / wt) of the total formulation amount. In yet another example, the solvent system contains a solvent modifying factor in an amount of 0.025% (wt / wt) of the total formulation amount. In yet another example, the solvent system contains a solvent modifying factor in an amount of 0.05% (wt / wt) of the total formulation amount. In yet another example, the solvent system contains a solvent modifying factor in an amount of 0.075% (wt / wt) of the total formulation amount. In yet another example, the solvent system contains a solvent modifying factor in an amount of 0.1% (wt / wt) of the total formulation amount. In yet another example, the solvent system contains a solvent modifying factor in an amount of 0.25% (wt / wt) of the total formulation amount. In yet another example, the solvent system contains a solvent modifying factor in an amount of 0.5% (wt / wt) of the total formulation amount. In yet another example, the solvent system contains a solvent modifying factor in an amount of 0.75% (wt / wt) of the total formulation amount. In yet another example, the solvent system contains a solvent modifying factor in an amount of 1.0% (wt / wt) of the total formulation amount. In yet another example, the solvent system contains a solvent modifying factor in an amount within the range of amounts described in this paragraph. The solvent modifying factor can be one or more selected from the group consisting of lemon oil, vitamin E, and methylsulfonylmethane (MSM).

[0127] In one example, the solvent system contains forskolin in an amount of 0.001% (wt / wt) of the total formulation amount. In another example, the solvent system contains forskolin in an amount of 0.0025% (wt / wt) of the total formulation amount. In yet another example, the solvent system contains forskolin in an amount of 0.005% (wt / wt) of the total formulation amount. In yet another example, the solvent system contains forskolin in an amount of 0.01% (wt / wt) of the total formulation amount. In yet another example, the solvent system contains forskolin in an amount of 0.015% (wt / wt) of the total formulation amount. In yet another example, the solvent system contains forskolin in an amount of 0.02% (wt / wt) of the total formulation amount. In yet another example, the solvent system contains forskolin in an amount of 0.025% (wt / wt) of the total formulation amount. In yet another example, the solvent system contains forskolin in an amount of 0.03% (wt / wt) of the total formulation amount. In yet another example, the solvent system contains forskolin in an amount of 0.035% (wt / wt) of the total formulation amount. In yet another example, the solvent system contains forskolin in an amount of 0.04% (wt / wt) of the total formulation amount. In yet another example, the solvent system contains forskolin in an amount of 0.045% (wt / wt) of the total formulation amount. In yet another example, the solvent system contains forskolin in an amount of 0.005% (wt / wt) of the total formulation amount. In yet another example, the solvent system contains forskolin in an amount of 0.055% (wt / wt) of the total formulation amount. In yet another example, the solvent system contains forskolin in an amount of 0.06% (wt / wt) of the total formulation amount. In yet another example, the solvent system contains forskolin in an amount of 0.065% (wt / wt) of the total formulation amount. In yet another example, the solvent system contains forskolin in an amount of 0.07% (wt / wt) of the total formulation amount. In yet another example, the solvent system contains forskolin in an amount of 0.075% (wt / wt) of the total formulation amount.

[0128] In one example, the solvent system contains a skin stabilizer in an amount of 0.01% (wt / wt) of the total formulation amount. In another example, the solvent system contains a skin stabilizer in an amount of 0.02% (wt / wt) of the total formulation amount. In yet another example, the solvent system contains a skin stabilizer in an amount of 0.03% (wt / wt) of the total formulation amount. In yet another example, the solvent system contains a skin stabilizer in an amount of 0.04% (wt / wt) of the total formulation amount. In yet another example, the solvent system contains a skin stabilizer in an amount of 0.05% (wt / wt) of the total formulation amount. In yet another example, the solvent system contains a skin stabilizer in an amount of 0.06% (wt / wt) of the total formulation amount. In yet another example, the solvent system contains a skin stabilizer in an amount of 0.07% (wt / wt) of the total formulation amount. In yet another example, the solvent system contains a skin stabilizer in an amount of 0.08% (wt / wt) of the total formulation amount. In yet another example, the solvent system contains a skin stabilizer in an amount of 0.09% (wt / wt) of the total formulation amount. In yet another example, the solvent system contains a skin stabilizer in an amount of 0.10% (wt / wt) of the total formulation amount. In yet another example, the solvent system contains a skin stabilizer in an amount of 0.15% (wt / wt) of the total formulation amount. In yet another example, the solvent system contains a skin stabilizer in an amount of 0.20% (wt / wt) of the total formulation amount. In yet another example, the solvent system contains a skin stabilizer in an amount of 0.25% (wt / wt) of the total formulation amount. In yet another example, the solvent system contains a skin stabilizer in an amount of 0.30% (wt / wt) of the total formulation amount. In yet another example, the solvent system contains a skin stabilizer in an amount of 0.35% (wt / wt) of the total formulation amount. In yet another example, the solvent system contains a skin stabilizer in an amount of 0.40% (wt / wt) of the total formulation amount. In yet another example, the solvent system contains a skin stabilizer in an amount of 0.45% (wt / wt) of the total formulation amount. In yet another example, the solvent system contains a skin stabilizer in an amount of 0.50% (wt / wt) of the total formulation amount. In yet another example, the solvent system contains a skin stabilizer in an amount within the range of amounts described in this paragraph.The skin stabilizer can be one or more selected from the group consisting of lauricidin (registered trademark), D-panthenol (dexpanthenol), and phytantriol.

[0129] Administration In some examples, the transdermal insulin formulation disclosed herein enables the direct delivery of insulin to the patient's body where insulin receptors are present in cells. Thus, the transdermal insulin formulation described herein achieves a bioavailability comparable to that by injection of insulin to a subject in need thereof.

[0130] In some examples, the transdermal insulin formulation disclosed herein is applied to any part of the skin, such as, for example, the plantar arch, the lateral heel, the palm, the upper arm, the ventral forearm, the dorsal forearm, the back, the chest, the thigh, the abdomen, the groin, the scalp, the axilla, the forehead, the lower back, the buttocks, etc. In these embodiments, the optimal sites for providing the transdermal insulin formulation disclosed herein are the ventral forearm, the upper arm, and the chest.

[0131] In some examples, the transdermal insulin formulation disclosed herein includes a liquid dosage form, such as, for example, a solution, a liquid spray, a lotion, etc.

[0132] In one example, the method of administering the formulation described herein includes using a spray device. The spray device can be a single-dose or multi-dose system and includes, for example, a bottle, a pump, and an actuator, and various commercially available ones are available. As an example, for a spray device, the typical volume of the liquid sprayed in a single spray operation is 0.01 ml, 0.02 ml, 0.03 ml, 0.04 ml, 0.05 ml, or 0.06 - 0.14 ml, such as 0.08 - 0.12 ml (0.1 ml, etc.).

[0133] In some instances, the transdermal insulin formulations disclosed herein can be designed for rapid release of insulin and transdermal absorption. In other instances, the transdermal insulin formulations disclosed herein can be designed for sustained release of insulin over time and transdermal absorption.

[0134] In some instances, the transdermal insulin formulations disclosed herein are administered in a single dose, whereby a defined amount of insulin is administered at one time. In other instances, the transdermal insulin formulations disclosed herein are administered by multiple doses at one or more divided doses over a specified time period.

[0135] In some instances, the transdermal insulin formulations disclosed herein can be adjusted for an individual patient in accordance with clinical symptoms and baseline blood glucose concentrations in the serum. In these embodiments, the transdermal pharmaceutical composition can be formulated with various concentrations of insulin and a suitable dosing regimen to more closely mimic the physiological pulsatile secretion of insulin, thereby maintaining serum glucose levels within the physiological range.

[0136] In one example, the transdermal insulin formulations described herein are administered within a dosage range of insulin from about 25 IU / day to about 500 IU / day.

[0137] In one example, a method for delivering insulin to a subject in need thereof is disclosed herein, the method comprising administering to the subject a therapeutically effective amount of the transdermal insulin formulation described herein.

[0138] In one example, a method for stabilizing glucose levels in a subject receiving insulin is disclosed herein, the method comprising administering to the subject in need thereof a therapeutically effective amount of the transdermal insulin formulation described herein.

[0139] In one example, a method for treating diabetes is disclosed herein, the method comprising administering to the subject in need thereof a therapeutically effective amount of the transdermal insulin formulation described herein.

[0140] In one example, a method comprising administering to a subject in need thereof a therapeutically effective amount of the transdermal insulin formulation described herein and further administering at least one additional therapeutic agent. The at least one therapeutic agent can be subcutaneous administration of another insulin and / or any other commercially available therapeutic agent for stabilizing glucose levels, stimulating natural insulin production, and / or treating diabetes in the subject. For example, such additional therapeutic agents include, but are not limited to, metformin and repaglinide.

[0141] In one example, the present disclosure provides a method for delivering at least 90% of at least one insulin rapidly through the skin to the lower adipose tissue, stroma, and capillary network. This delivery can be achieved in just a few tens of seconds or less than a few minutes.

[0142] The present application also discloses a method for preparing a transdermal insulin formulation as described herein. In one example, the method includes: (a) Selecting at least one insulin; (b) Determining an effective dose of the at least one insulin, wherein the effective dose of the at least one insulin has molecular properties including van der Waals forces and dipole moments; (c) Quantifying the molecular properties of the at least one insulin; (d) Determining the amount of a solvent system for solubilizing the effective dose of the at least one insulin, wherein the amount of the solvent system has molecular properties including van der Waals forces and dipole moments; (e) Quantifying the molecular property amount of the amount of the solvent system; (f) Comparing the molecular properties of the at least one insulin with the molecular properties of the solvent system; (g) Determining that the molecular characteristics of the insulin-free solvent system are substantially the same as or approximately ±20% of the molecular characteristics of at least one type of insulin; and (h) Combining the solvent system and the at least one type of insulin to provide a transdermal insulin formulation.

[0143] In one example, a method for preparing a transdermal insulin formulation as described herein includes selecting one or more components for the solvent system to determine the amount of the solvent system to solubilize the effective dose of the at least one type of insulin, the one or more components being selected from the group consisting of two or more solvents, solvent modifiers, solute modifiers, sources of cell activation energy, skin stabilizers, and combinations thereof; each of the one or more components having molecular characteristics including van der Waals forces and dipole moments.

[0144] In another example, a method for preparing a transdermal insulin formulation as described herein includes selecting one or more components for the solvent system to determine the amount of the solvent system to solubilize the effective dose of the at least one type of insulin, the one or more components being selected from the group consisting of two or more solvents, solvent modifiers, solute modifiers, sources of cell activation energy, skin stabilizers, one type of membrane permeability regulator, at least one type of enzyme activator, at least one type of capillary dilator, and combinations thereof; each of the one or more components having molecular characteristics including van der Waals forces and dipole moments.

[0145] In one example, a method of selecting components and amounts for preparing a transdermal insulin formulation as described herein is disclosed herein, where the method includes the following steps: (a) Selecting at least one type of insulin necessary to treat a particular condition; (b) Quantifying the amount of the insulin for the effective dose; (c) Quantifying the molecular properties of the insulin as described above to include the sum of van der Waals forces and molar moments; (d) Searching for a solvent for the insulin as described above; (e) Quantifying the amount of the solvent to solubilize the insulin as described above; (f) Quantifying the molecular properties of the solvent as described above to include van der Waals forces and dipole moments; (g) Comparing the molecular properties of the solvent as described above with the molecular properties of the insulin as described above; (h) Determining additional components to form a solvent system for the transfer; (i) Quantifying the molecular properties of the additional components as described above to include van der Waals forces and molar moments; (j) Determining the weighted sum of the molecular properties of the additional components as described above and the molecular properties of the solvent as described above to determine the molecular properties of the solvent system as described above; (k) Summing the molecular properties of the solvent system and the insulin as described above; (l) Comparing (j) and (k); and (m) Adjusting the delivery system such that the molecular properties of the at least one type of insulin in the solvent system are substantially the same as the molecular properties of the solvent system without insulin.

[0146] In one example described herein, the van der Waals forces and / or dipole moments of at least one type of insulin in the delivery system are approximately ±20% of the van der Waals forces and / or dipole moments of the solvent system. In one example described herein, the van der Waals forces and / or dipole moments of at least one type of insulin in the delivery system are approximately ±15% of the van der Waals forces and / or dipole moments of the solvent system. In one example described herein, the van der Waals forces and / or dipole moments of at least one type of insulin in the delivery system are approximately ±10% of the van der Waals forces and / or dipole moments of the solvent system. In one example described herein, the van der Waals forces and / or dipole moments of at least one type of insulin in the delivery system are approximately ±5% of the van der Waals forces and / or dipole moments of the delivery system.

[0147] Transdermal insulin formulations as described herein were continuously prepared and tested in a single patient described as having unstable type 2 diabetes (T2D) with low insulin sensitivity. These experiments showed that the transdermal insulin formulations disclosed herein: (a) can be transdermally supplemented on a one-to-one basis with injectable dosage forms; (b) can be delivered as immediate-acting, long-acting, and Humulin without difference in results, avoiding the need for syringes; (c) provide a more level insulin profile, minimizing bias; (d) avoid hypoglycemia tendencies by localizing receptor effects at the site of application when used exclusively; and (e) transiently enhance insulin sensitivity even for injection forms was demonstrated.

[0148] The following examples are intended to specifically illustrate the scope of the disclosure and not to limit it. It should be understood that other formulations known to those skilled in the art may be used as alternatives.

[0149] Example Example 1: Process for the Preparation of a Transdermal Insulin Preparation Weigh the solvent (anhydrous ethanol, polyethylene glycol or carbonate and acetone), and mix in a reaction vessel at ambient temperature with moderate stirring.

[0150] Weigh the excipients, grind the two crystalline forms of glycerol monolaurate and MSM into powder to accelerate dissolution, and mix into the reaction vessel.

[0151] Weigh and add the other liquid and semi-solid excipient components, and mix into the reaction vessel except for phosphoric acid.

[0152] Weigh and add phosphoric acid, and mix into the reaction vessel.

[0153] Add an appropriate weight of insulin to the reaction vessel.

[0154] Mix for approximately 1 hour until the solution becomes clear.

[0155] The transdermal insulin preparation thus prepared was used in the amounts indicated in the examples described below. Unless otherwise specified, all components are of USP grade. The insulin referred to in this specification is any form of insulin including rapid-acting, short-acting, intermediate-acting, and / or long-acting insulin.

[0156] Example 2: Formulation A transdermal insulin preparation was prepared using the above process comprising insulin and the solvent system described herein. An example of such a formulation is highlighted in Figure 24. The ranges of amounts considered for each component of the formulation are set forth in Table 2 below.

[0157] [Table 2]

[0158] The insulin titer of the obtained preparation was in the range of 10 IU / ml to 1600 IU / ml. Three preparations having an insulin titer of 200 IU / mL were selected for the clinical trial.

[0159] Example 3: Clinical Trial The preparation described in Example 2 was used in a single patient clinical trial. The basic clinical protocol was as follows: (a) A single dose of Novolog® was administered subcutaneously at approximately 6 am to adjust the blood glucose level to the targeted level at 9 am (about 100 mg / dL); (b) In one exception, Lantus® (long-acting basal insulin) was used as background support; when a rapid decline in the Lantus effect was observed around 2 pm, the Lantus dose was split into two parts (BID) per day, and at that time, the post-meal spike was reduced and the leveling was continued; (c) The first transdermal dose was administered approximately at 9 am every day. (d) Transdermal insulin was sprayed and applied to the inner forearm or chest using a metered-dose (0.2 mL / pump) finger-actuated nebulizer; and (e) The blood glucose level was measured at the pre-dose baseline and then every hour thereafter.

[0160] Experiment 1 The preparation delivered insulin at 209 IU / mL to evaluate performance. Blood glucose was monitored from 3 days before to 3 days after. Insulin was administered transdermally (TD) approximately every 3 to 4 hours for 15 hours after meals to patients on a 24-hour fast. Blood glucose was maintained at approximately 120 mg / dL until 1 hour after meals (10 hours after TD administration). Blood glucose began to rise when TD administration was reduced and returned to the pre-TD value 12 hours after TD administration. As can be seen from Figure 1 and Table 3 below, the value of insulin sensitivity after TD administration was higher than that before administration until 50 hours (the last measurement).

[0161]

Table 3

[0162] Experiment 2 The formulation delivered insulin at 201 IU / mL to evaluate performance. Blood glucose was also monitored from 3 days before to 3 days after. Transdermal insulin was administered in large dose boluses (201 IU) approximately every 3 - 8 hours for 15 hours after a meal. Blood glucose was maintained within approximately ±20 mg / dL of the value before transdermal administration throughout 3 days when lunch was omitted (12 - hour fasting). Blood glucose began to decline when transdermal administration was reduced and the administration of Novolog® was increased, and was still lower than before transdermal administration for 72 hours. As can be seen from Figure 2 and Table 4 below, the values of insulin sensitivity after transdermal administration were higher than before administration until 50 hours (the last measurement performed).

[0163]

Table 4

[0164] Experiment 3 The formulation delivered insulin at 209 IU / mL to evaluate performance. Blood glucose was monitored from 3 days before to 3 days after. Transdermal insulin was first administered at 9:02 and was administered for 14 hours after a meal using large dose boluses (209 IU) approximately every 2 - 6 hours. Novolog® was co - administered to adjust blood glucose throughout transdermal administration. As can be seen from Figure 3 and Table 5 below, the values of insulin sensitivity after transdermal administration were higher than before administration until 50 hours (the last measurement performed).

[0165]

Table 5

[0166] Data Analysis Novolog (registered trademark) and regular insulin (Humulin (registered trademark)) are metabolized in 4 hours and 8 hours respectively and have peak availability at 2 hours and 4 hours. The insulin delivery curve is similar to a normal distribution and thus gives a reasonable prediction of insulin availability using ±2σ AUC.

[0167] Predicted blood glucose The predicted value of the glucose increase per hour was calculated by summing the contributions of Lantus (registered trademark) (Table 6 below), the hourly contributions of Novolog (registered trademark) and human insulin, using a default insulin sensitivity of 2 mg / dL / IU, and comparing it with the blood glucose observed 1 hour earlier.

[0168]

Table 6

[0169] Insulin sensitivity Insulin sensitivity was estimated throughout by comparing the predicted blood value to the actual blood glucose value measured at that time; for example, T1 = 200 mg / dL, 80 IU Novolog (registered trademark) SC x 2 mg / dL / IU = Δ - 160 mg / dL, +27 mg / dL / hour x 4 hours = Δ108 mg / dL, predicted blood glucose = 148.

[0170] Therefore, the resulting sensitivity value is a composite value of Novolog (registered trademark) and human insulin relative to the default insulin sensitivity. Values higher than 2 indicated typically greater insulin sensitivity. For SC delivery and TD delivery, sensitivity was tabulated in each individual experiment and considered in the experimental results described below.

[0171] Novolog® was typically administered to bring morning values into the desired target range (90 - 120 mg / dL), so sensitivity was calculated during the morning and during the experiment if possible. The sensitivity values for Novolog® were tabulated in Table 7 and graphed in Figure 4. The overnight sensitivity of Novolog® was also calculated, daily, after TD administration (without food or insulin). This overnight sensitivity calculation included the contribution of Lantus® as the only correction factor (Table 6 below), and thus was as close as possible, under these experimental conditions, to a "stand alone" estimate (post - meal correction not required).

[0172]

Table 7

[0173] The calculation of insulin sensitivity was performed with a conservative bias for the transdermal dose throughout the experiment. "Selected Average" was reported, which excluded the points at which the prediction model began to deviate significantly from the actual values (e.g., after large consecutive bolus doses at each time point, large post - meal spikes). Based on this modeling, decreased insulin sensitivity reflects the increased insulin present in this system.

[0174] In summary, the inventors modeled the normal delivery curves for both the subcutaneous dosage form and the transdermal formulation of Novolog®.

[0175] Results In Experiment 1, the subjects were dosed in pulses of 40 - 80 IU while fasting. The last meal was approximately 7 PM the day before transdermal administration, and dosing was started at 8:55 AM. Blood glucose, which should have increased at an accelerated pace due to fasting, remained flat throughout the day.

[0176] Experiment 2 was administered repeatedly as a 200 IU bolus. Performance was as expected, with a normalized prolongation and significantly increased insulin sensitivity after transdermal administration.

[0177] Experiment 3 was a reverse paradigm, examining the effect of transdermal administration in combination with subcutaneous administration. While Novolog® was given throughout the day, transdermal human insulin was administered repeatedly as a 200 IU bolus. The dose of Novolog® was substantially lower than the dose required in the absence of the transdermal formulation to obtain the desired blood glucose levels. Blood glucose decreased to 113 within 2 hours of transdermal administration and remained below pre - transdermal levels over 72 hours.

[0178] Furthermore, in Experiment 3, Novolog® was administered at 6:49 am at the end of a 48 - hour period of subcutaneous administration of short - acting and long - acting insulin, and at 10:00 am, 12:00 pm, and 2:00 pm by transdermal administration. Despite the fact that the effect of Novolog® may have been approximately 85% exhausted by 10:45 pm, the levels set by Novolog® showed a downward trend with the introduction of transdermal insulin, maintained post - meal spikes low, then decreased the levels to approximately the baseline at 6:49 am, confirming that the introduction of transdermal insulin to the skin makes the insulin stored in the subcutaneous adipose tissue available for use.

[0179] Classification insulin sensitivity results: Normalization was observed at lower formulation strengths, and these values were in line with the expected 8 - hour availability of transdermal insulin administration. This normalization suggests that excess transdermal insulin is retained and available for subsequent use. The subcutaneous sensitivity of Novolog® was generally higher in the morning than the insulin delivered by the transdermal formulation (2.1 and 1.8 on average, respectively). These overnight values were 3.1 on average, in contrast to 2.3 at the pre - test time point.

[0180] Insulin sensitivity is a measure of how well the body utilizes the supplied insulin. Generally, this value decreases with time (the age of the subject) and is generally treated as a long-term average. To understand the transdermal system, Novolog® sensitivity was measured before, during, and after the experiment and compared to the assumed sensitivity of 2. Table 7 and Figure 4 show Novolog® sensitivity before overnight and before transdermal insulin delivery and, if available, overnight during the withdrawal period of transdermal administration. The low points in Table 7 (sensitivity ≤ 2.1) were as follows: out of 48 data points, 16 data points ≤ 2.1, with an average sensitivity of 3.33. (a) Morning, after breakfast (b) Afternoon, after lunch (c) Evening, after dinner (d) Overnight, after a 4-day withdrawal period in transdermal administration (e) Overnight, during the experimental period without overnight Lantus® administration (f) Novolog® sensitivity was thus generally much higher than before the transdermal experiment and substantially higher overnight. Since none of the treatments showed the ability to increase insulin sensitivity, this effect is considered to be due to the contribution from the retained transdermal insulin rather than a true shift in the subject's insulin sensitivity. (g) The values of Novolog® sensitivity in the morning (after breakfast) are necessarily more variable since the contribution of the meal is not incorporated. The values range from 0.41 to 4.83 and have an average of 2.18, which is consistent with the values before the transdermal experiment.

[0181] Total insulin sensitivity As shown in the above experiment, Novolog® and transdermal insulin produced equivalent results. The calculation of total insulin sensitivity includes the combined effect of both subcutaneous and transdermal when subcutaneous insulin is incorporated during the transdermal experiment.

[0182] Example 4: Analysis of the Permeation of the product through the artificial skin model in the support medium Overview of the artificial skin model preparation To produce a dermal equivalent (DE), primary adult skin fibroblasts were embedded in a fibrin matrix. The DE was cultured so that the fibroblasts could reconstruct the matrix. Primary human neonatal keratinocytes were applied to the surface of the DE and cultured under liquid for 48 hours. The artificial skin model was cultured at the air / liquid interface (ALI) until a stratified epidermis was formed. The incubation conditions for all cultures were 5 ± 1% (v / v) CO 2 2, at 37 ± 2 °C, with a relative humidity (RH) ≥ 95%.

[0183] Permeation study of a transdermal formulation of 100 IU / ml versus injectable insulin in the artificial skin model construct Plates with shallow wells containing the artificial skin model were prepared by adding 1 mL of freshly warmed maintenance medium to each well. 11 μL of the transdermal formulation as described in this application at 100 IUs / ml, injectable insulin, or PBS as a blank control was applied to the surface of the artificial skin model. Collection of the supernatant was performed at time points of 0, 3, 5, 10, 15, 20, 25, 30, 40, 50, and 60 minutes.

[0184] ELISA analysis of insulin in the supernatant was performed using a human insulin ELISA kit (R&D systems DINS00). Back-calculation of the concentration was performed using MyCurveFit software.

[0185] The average OD values of permeation for the test transdermal formulation, injectable insulin, or PBS control are shown in Table 8 below.

[0186]

Table 8

[0187] A histogram of the OD values as highlighted in Table 1 above is shown in Figure 5. The permeation of the transdermal formulation versus injectable insulin through the artificial skin model is shown as a graph in Figure 6.

[0188] The back-calculated concentrations of insulin recovered in the supernatants after permeation through the artificial skin model are shown in Table 9 below.

[0189]

Table 9

[0190] The interpolated concentrations of the transdermal formulation relative to injectable insulin and the total insulin recovery amounts at each time point are shown as a graph in Figure 7.

[0191] Controls of medium only and insulin (high, medium, and low control sets from Bio-Techne catalog number QC107) were also measured on ELISA plates. In addition to the medium-only and insulin control sets, standard OD values for various concentrations of insulin are shown in Table 10 below.

[0192]

Table 10

[0193] Figure 8 shows a graph of the insulin ELISA standard curve. Using a linear plot, an R2 value of 0.9996 was obtained. The OD values for the test items were interpolated against the standard curve plot to generate pMol / L values.

[0194] The above data indicate that (i) the values for the transdermal formulation were higher than those for injectable insulin at all time points, (ii) peak concentrations were observed at the 3-minute time point and there was another peak at the 60-minute final time point, and (iii) the amount of insulin recovered for the transdermal formulation was twice that for injectable insulin.

[0195] Immunohistochemical analysis The artificial skin model constructs were bisected and cut into 6-μm thick cryosections at -25°C. The sections were placed on adhesive slides, air-dried, and fixed in 100% ethanol before staining. Staining of cell structures was performed with hematoxylin and eosin (H&E). Visualization of insulin in the artificial skin model was carried out by using an anti-insulin antibody (mouse monoclonal antibody produced against human insulin, Abcam 133289). The sections were incubated with the antibody diluted at a concentration of 0.016 μg / ml for 1 hour at room temperature. The antibody was diluted with a diluent / blocker (SP-5035) from Vector laboratory (USA). An indirect system was used to amplify the signal, and a DAB chromogen (brown) was used as the endpoint (PK-8200 from Vector Laboratories). All sections were microphotographed at a magnification of x200 with a Leica microscope.

[0196] Figures 9, 10, and 11 show the staining of the artificial skin model constructs, where fibroblasts and keratin-producing cells in the constructs are stained purple, and the test insulin samples (i.e., transdermal insulin formulations and injectable insulin) are stained brown. Figures 9A, 9B, 9C, 9D, 9E, and 9F show the staining of control constructs at various time points including 0 minute, 5 minutes, 10 minutes, 20 minutes, 40 minutes, and 60 minutes, respectively. Figures 10A, 10B, 10C, 10D, 10E, and 10F show the staining of injectable insulin-treated constructs at various time points including 0 minute, 5 minutes, 10 minutes, 20 minutes, 40 minutes, and 60 minutes, respectively. Figures 11A, 11B, 11C, 11D, 11E, and 11F show the staining of transdermal insulin composition-treated constructs at various time points including 0 minute, 5 minutes, 10 minutes, 20 minutes, 40 minutes, and 60 minutes, respectively.

[0197] The above immunohistochemical analysis of the artificial skin model constructs shows the translocation of insulin (both injectable and transdermal) from the stratum corneum through the epidermis and dermis.

[0198] The results obtained by permeation and immunohistochemical analysis demonstrate faster movement with the transdermal formulation compared to injectable insulin.

[0199] Example 5: Transdermal Insulin (TD) Dose-Response Study in Control Male Rats The following study was designed to examine the time-course dose-response effects of skin application of various transdermal formulations on fasting blood glucose in control male Wistar rats. Blood samples were also collected at the same time points after treatment to measure plasma insulin levels.

[0200] Method Preparation of Rat Skin On day 0 (24 hours before the first test day), a small patch (2x3 cm) of the dorsal hair was shaved using appropriate scissors. The area of "naked" skin was then thoroughly cleaned with cold water and gently patted dry with a dry swab. The animal was returned to the breeding cage and allowed to recover for 24 hours.

[0201] Application of Transdermal Insulin On the test day (day 1), food was removed and the animal was transferred to a clean cage with free access to tap water. After 4.5 hours and before treatment, the baseline fasting blood glucose was measured from the tip of the tail using an Accu-Chek glucometer, and 100 μl of blood was collected into an EDTA Microvette® tube, stored on ice before centrifugation, and then the plasma was collected and stored at -20°C. An insulin solution or vehicle was gradually applied to the skin of the rats prepared at 1 ml per kg body weight using a 1 m disposable plastic syringe. At the same time, the test material was gently rubbed into the skin using the index finger of the gloved hand. More specifically, the index finger was first rotated 10 times clockwise and then 10 times counterclockwise.

[0202] Blood Glucose Measurement Blood glucose was measured at 5, 10, 20, 40, 60, 90, and 120 minutes after application of the insulin solution or vehicle using an Accu-Chek Active glucometer and glucose test strips.

[0203] Blood Sampling and Plasma Preparation Blood samples were collected from the cut end of the tail after application of lignocaine gel (Biorex Laboratories UK). 100 μl of blood was collected at each of the above time points and processed as described above. Blood collection was facilitated by placing the animals in a warm environment (25 °C). This has no harmful effects on the animals. Blood was collected into EDTA-coated Microvette® tubes (Sarstedt microvette CB 300 LH, reference number: 16.443, Aktiengesellschaft & Co., D-51588, Numbrecht, Germany) for measurement of plasma insulin concentration, placed on ice and then centrifuged at approximately 500 x G for 5 minutes. The resulting plasma was stored at -20 °C until required. Multiple freeze / thaw cycles were avoided.

[0204] Plasma Insulin Measurement The levels of human insulin measured in rat plasma samples were measured using the Crystal Chem human insulin ELISA kit (Catalog number: 90095). The Crystal Chem human insulin ELISA kit is an ELISA sandwich assay for human insulin. This utilizes specific antibodies immobilized on the wells of a microplate. Briefly, 100 μl of HRP-labeled human insulin antibody and 25 μl of standard or plasma sample were added to each well. After incubation at 37 °C for 2 hours, the plate was washed 3 times with 300 volumes of wash buffer, then 100 μl of HRP substrate solution was added to each well and incubated in the dark at room temperature for 15 minutes. The enzyme reaction was stopped by adding 100 μl of stop solution and the plate was read at two wavelengths, 450 and 630, using a Spectra Max250 (Molecular Devices, San Jose, California 95134). The results were converted to insulin values using a human insulin standard.

[0205] Results The results for individual blood glucose levels are presented in a figure as follows.

[0206] Figure 12 shows blood glucose levels measured after a 4.5-hour fast using an Accu-Chek® Active glucose meter and glucose test strips before and after application of 1 ml / kg of vehicle (placebo) at 5, 10, 20, 40, 60, 90, and 120 minutes.

[0207] Figure 13 shows blood glucose levels measured after a 4.5-hour fast using an Accu-Chek® Active glucose meter and glucose test strips before and after application of human insulin solution (0.1 IU / kg / ml) at 5, 10, 20, 40, 60, 90, and 120 minutes.

[0208] Figure 14 shows blood glucose levels measured after a 4.5-hour fast using an Accu-Chek® Active glucose meter and glucose test strips before and after application of human insulin solution (0.2 IU / kg / ml) at 5, 10, 20, 40, 60, 90, and 120 minutes.

[0209] Figure 15 shows blood glucose levels measured after a 4.5-hour fast using an Accu-Chek® Active glucose meter and glucose test strips before and after application of human insulin solution (0.4 IU / kg / ml) at 5, 10, 20, 40, 60, 90, and 120 minutes.

[0210] Figure 16 shows blood glucose levels measured after a 4.5-hour fast using an Accu-Chek® Active glucose meter and glucose test strips before and after application of human insulin solution (0.8 IU / kg / ml) at 5, 10, 20, 40, 60, 90, and 120 minutes.

[0211] Figure 17 shows blood glucose levels measured after a 4.5-hour fast using an Accu-Chek® Active glucose meter and glucose test strips before and after application of human insulin solution (1.6 IU / kg / ml) at 5, 10, 20, 40, 60, 90, and 120 minutes.

[0212] Figure 18 shows data presented as the percentage change in blood glucose levels relative to the baseline value (time 0) after transdermal application of human insulin solution (0.1 IU / kg / ml) or vehicle (1 ml / kg).

[0213] Figure 19 shows data presented as the percentage change in blood glucose levels relative to the baseline value (time 0) after transdermal application of human insulin solution (0.2 IU / kg / ml) or vehicle (1 ml / kg). Statistical analysis was performed using one-way ANOVA, followed by Dunnett's multiple comparison test. Statistical significance is represented as *p < 0.05.

[0214] Figure 20 shows data presented as the percentage change in blood glucose levels relative to the baseline value (time 0) after transdermal application of human insulin solution (0.4 IU / kg / ml) or vehicle (1 ml / kg). Statistical analysis was performed using one-way ANOVA, followed by Dunnett's multiple comparison test. Statistical significance is represented as *p < 0.05 and **p < 0.01.

[0215] Figure 21 shows data presented as the percentage change in blood glucose levels relative to the baseline value (time 0) after transdermal application of human insulin solution (0.8 IU / kg / ml) or vehicle (1 ml / kg). Statistical analysis was performed using one-way ANOVA, followed by Dunnett's multiple comparison test. Statistical significance is represented as *p < 0.05.

[0216] Figure 22 shows data presented as the percentage change in blood glucose levels relative to the baseline value (time 0) after transdermal application of human insulin solution (1.6 IU / kg / ml) or vehicle (1 ml / kg). Statistical analysis was performed using one-way ANOVA, followed by Dunnett's multiple comparison test. Statistical significance is represented as *p < 0.05.

[0217] Figure 23 shows the levels of human insulin measured in cumulative plasma samples collected from time 0 (before treatment) to 120 minutes after treatment. Results are the mean ± standard error of two values for vehicle-treated rats and four values for 0.2 IU and 0.4 IU / kg / ml treated animals. Statistical significance is represented as ***p < 0.001 and was determined using one-way analysis of variance followed by Dunnett's multiple comparison test against the vehicle-treated group. Statistical significance was represented as ***p < 0.001 and the 0.2 IU and 0.4 IU / kg / ml insulin treatment groups were compared using Student's t-test.

[0218] Conclusion The above data showed that TD insulin crossed the skin barrier and was detected in rat plasma in a dose-response manner.

[0219] The above data also showed that the transdermal insulin delivery route had a reduced effect on fasting blood glucose in Wistar rats compared to placebo or vehicle-treated animals. Data on the time course of human insulin values after crossing the skin barrier could not be obtained; nevertheless, cumulative human levels were detected in plasma samples from treated rats. This supports that the transdermal delivery system acts as a vector or carrier that enables human insulin to cross the skin barrier and thereby lower blood glucose levels.

[0220] The foregoing description of specific embodiments is to disclose the overall nature of the embodiments described herein. By using current knowledge, others can easily make changes and / or modifications to such specific embodiments for various purposes without departing from the similar concept. Therefore, such modifications and changes should be understood to be within the intention and scope equivalent to the embodiments disclosed herein and are also so intended. It should be understood that the expressions or technical terms used herein are for the purpose of description and not for limitation. Therefore, although the embodiments described herein are described with preferred embodiments, those skilled in the art will understand that the embodiments described herein can be implemented with changes within the spirit and scope of the embodiments described herein.

Claims

1. (i)Insulin; and (ii)A solvent system comprising at least one selected from the group consisting of at least two solvents, at least one solvent modifying factor, at least one solute modifying factor, at least one source of cell activation energy, and at least one skin stabilizer, A transdermal insulin preparation comprising: The solvent system comprises molecular properties substantially similar to or approximately ±20% of the molecular properties of insulin in the preparation, and the molecular properties are selected from van der Waals forces and dipole moments, A transdermal insulin preparation.

2. (a)At least two solvents are present in an amount in the range of about 5% to about 90% of the preparation, (b)At least one solvent modifying factor is present in an amount in the range of about 0.0001% to about 50% of the preparation, (c)At least one solute modifying factor is present in an amount in the range of about 0.003% to about 5% of the preparation, (d)At least one source of cell activation energy is present in an amount in the range of about 0.01% to about 0.1% of the preparation, and (e)At least one skin stabilizer is present in an amount in the range of about 0.05% to about 5% of the preparation, The preparation of Claim 1.

3. The preparation of Claim 1, wherein the solvent system further comprises at least one selected from the group consisting of a membrane permeability regulating factor, an enzyme activating factor, and a capillary dilation factor.

4. The solvent system is (a)A membrane permeability regulating factor in an amount in the range of about 0.01% to about 5% of the preparation, (b)An enzyme activating factor in an amount in the range of about 0.01% to about 0.05% of the preparation, and (c)A capillary dilation factor in an amount in the range of about 0.1% to about 2% of the preparation, The preparation of Claim 1, further comprising at least one of the above.

5. The preparation of Claim 1, wherein the insulin is selected from the group consisting of rapid-acting insulin, short-acting insulin, intermediate-acting insulin, long-acting insulin, and mixtures thereof.

6. The preparation of Claim 1, wherein the solvent system comprises propylene carbonate and / or propylene glycol.

7. The preparation of Claim 1, wherein the solvent system comprises ethanol, propylene carbonate and / or propylene glycol, acetone, and phosphoric acid or any other acid.

8. The preparation of Claim 6, wherein phosphoric acid or any other acid is present in an acid-to-insulin molecule ratio in the range of 0.2 to 2.

0.

9. The preparation of Claim 1, formulated in a liquid dosage form.

10. The formulation of claim 1 which is in liquid dosage form, wherein the liquid dosage form ranges from 0.2 mL to 1 mL and contains insulin in an amount ranging from 7 IU / mL to 1,700 IU / mL.

11. The formulation of claim 1, wherein the solvent system comprises ethanol, propylene carbonate and / or propylene glycol, acetone, lemon oil, vitamin E, phytantriol, dexpanthenol, lauricidin, methylsulfonylmethane (MSM), forskolin, and phosphoric acid.

12. A process for preparing the formulation of claim 1, comprising: (a) selecting insulin; (b) determining an effective dose of insulin, wherein the effective dose of insulin has molecular properties including van der Waals forces and dipole moments; (c) quantifying the molecular properties of insulin; (d) determining an amount of a solvent system for solubilizing the effective dose of insulin, wherein the amount of the solvent system has molecular properties including van der Waals forces and dipole moments; (e) quantifying the amount of the molecular properties of the amount of the solvent system; (f) comparing the molecular properties of insulin with the molecular properties of the solvent system; (g) adjusting the molecular properties of the solvent system such that they are substantially the same as or approximately ±20% of the molecular properties of insulin; and (h) combining the solvent system and insulin to provide a transdermal formulation. A process comprising the above steps.

13. A method of delivering insulin to a subject in need thereof, comprising administering a therapeutically effective amount of the transdermal formulation of claim 1 to the subject.

14. A method of stabilizing glucose levels in a subject receiving insulin, comprising administering a therapeutically effective amount of the transdermal formulation of claim 1 to the subject in need thereof.

15. A method disclosed herein for delivering insulin while minimizing hypoglycemia, comprising administering a therapeutically effective amount of the formulation described herein to a subject in need thereof.

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