GLUCAGON COMPOSITION OR A GLUCAGON ANALOGUE FOR USE IN THE TREATMENT OF EXERCISE-INDUCED HYPOGLYCEMIA.

Abstract

Certain modalities refer to methods for the treatment, mitigation, or prevention of exercise-induced or exercise-associated hypoglycemia, which comprise the administration to a subject in need of a glucagon formulation or formulations, or a glucagon analogue, in an amount effective for the treatment, mitigation, or prevention of the condition. The subject may be administered a glucagon or glucagon analogue composition 1, 5, 10, 15 to 20, 25, or 30 minutes before the start of exercise.

Description

TREATMENT OF EXERCISE-INDUCED HYPOGLYCEMIA IN TYPE 1 AND TYPE 2 DIABETES IN WHICH INSULIN IS USED Cross-reference to Related Applications This application claims the benefit of United States Provisional Patent Application Number 61 / 609,964, filed on December 22, 2017, the contents of which are incorporated herein by reference. Background of the invention A. Field of invention The invention relates generally to compositions and methods connected with physiology and medicine. In particular, the compositions and methods relate to the treatment of a certain instance of hypoglycemia. B. Description of Related Technique Exercise is a cornerstone of type 1 diabetes (T1D) management (Chu et al., Physiol Sports Med 39: 64-77, 2011). However, blood glucose stability during exercise, and for up to 12 to 24 hours during recovery, remains a significant challenge (West et al., J Sports Sci 28: 781-88, 2010). The fear of hypoglycemia discourages many patients from engaging in aerobic exercise (Brazeau et al., Diabetes Care 11: 210-89, 2008). For those who choose to exercise regularly, hypoglycemia is a common complaint that frequently necessitates interruptions in sports and competitions, games, and training (Chu et al., Physiol Sports Med 39: 64-77, 2011).In order to reduce the incidence of hypoglycemia during and immediately after exercise, patients are advised to reduce their bolus dose in the meal preceding exercise by 25 to 75 percent (Chu et al., Physiol Sports Med 39: 64-77, 2011); however, this approach frequently results in pre-exercise hyperglycemia, particularly if exercise is performed 2 hours or more after the meal (West et al., J Sports Sci 28: 781-88, 2010; Rabasa-Lhoret et al., Diabetes Care 24: 625-30, 2001; Campbell et al., Diabetes Care 36: 2217-45, 2013). Even exercise in the fasted state in patients on continuous subcutaneous insulin infusion (CSII) promotes a drop in blood glucose if baseline insulin levels are mismatched (Stenerson et al., J Diabetes Sci Technol. PMID: 25231116, 2014). Nocturnal hypoglycemia after exercise is also very common in T1D, with approximately 50 percent of young patients developing the condition about 7 to 11 hours after the end of vigorous afternoon exercise (Tsalikian et al., J Psdiatr. 4: 528-34, 2005). If insulin is administered to correct food-related post-exercise hyperglycemia (often referred to as rebound hyperglycemia), severe post-exercise hypoglycemia can occur, which can even result in death (Tanenberg et al., Endocr Pract. 16(2): 244-8, 2010).Patients may reduce the dose of rapid-acting insulin administered with the dinner meal after late-night exercise in order to reduce the risk of nocturnal hypoglycemia (Campbell et al., PLoS One 9(5): e97143, 2014) or they may reduce the supply of basal insulin for 6 hours at bedtime (Taplin et al., J Pediatr 157(5): 784-8, 2010) in order to help mitigate the risk, but these strategies frequently result in hyperglycemia (Campbell et al., PLoS One 9(5): e97143, 2014; Taplin et al., J Pediatr 157(5): 784-8, 2010). Although extra carbohydrate intake not covered by insulin administration may help prevent hypoglycemia during and after exercise (Riddeil et al., Int J Sport Nutr.9 (1): 24-34, 1999), excessive intake negates the patient's ability to have a negative caloric balance, thereby limiting the patients' ability to maintain or lose their body weight. Patients on insulin pump therapy have the flexibility to reduce basal insulin delivery in anticipation of exercise and during recovery to help guard against hypoglycemia. Guidelines from the International Society for Pediatric and Adolescent Diabetes recommend that basal insulin reductions should be made 60 to 90 minutes before the start of exercise so that circulating insulin levels are lowered at the onset of activity (Robertson et al., Pediatr Diabetes. 15 Suppl 20: 203-23, 2014). This is a somewhat problematic and unpredictable task that is not usually carried out correctly, even by the most educated and motivated patients. This is because glucose production by the liver during moderate-intensity exercise is primarily facilitated by an increase in the glucagon-alpha ratio.insulin (Camacho et al., Exerc Sport Sci Rev. 33 (1): 17-23, 2005), since patients with T1D have a reduction in this ratio during exercise due to relative peripheral hyperinsulinemia (Camacho et al., Exerc Sport Sci Rev. 33 (1): 17-23, 2005) and insufficient glucagon secretion (Oskarsson et al., Diabetes Metab. 25 (6): 491-714, 1999), it may be better to try to change this ratio by administering glucagon at the beginning of exercise. Brief Description of the Invention The compositions and methods of the present invention provide a solution to the glycemic control problems associated with exercise in diabetic subjects. In particular, glucagon or a glucagon analogue is administered before, during, and after exercise; before and during; before and after; or before, during, and after exercise. In certain aspects, glucagon or a glucagon analogue is administered before exercise and optionally during exercise. By way of example, the inventors have discovered a process for the treatment, mitigation, or prevention of exercise-induced hypoglycemia while preventing hyperglycemia. Certain modalities refer to methods for the treatment, mitigation, or prevention of exercise-induced or exercise-associated hypoglycemia, which comprise the administration to a subject in need of a glucagon formulation or formulations, or a glucagon analogue (e.g., dasiglucagon), in an amount effective for the treatment, mitigation, or prevention of the condition. A glucagon or glucagon analogue composition may be administered to the subject at 1.5, 10, 15, 20, 25, or 30 minutes, including all values ​​and intervals thereto, prior to the start of exercise. In certain aspects, the subject is a diabetic subject. In an additional aspect, 25 pg, 50 pg, 75 pg, 100 pg, 125 pg, 150 pg, 175 pg, 200 pg, 225 pg, 250 pg, or 275 pg to 300 pg of glucagon or a glucagon analogue are administered, and in certain aspects 150 pg ± 50 pg.Glucagon or a glucagon analogue can be administered as a bolus or as an infusion over 5 to 30 minutes. In some cases, a second dose of glucagon or a glucagon analogue is administered during exercise. The second dose may be 25 pg, 50 pg, 75 pg, 100 pg, 125 pg, 150 pg, 175 pg, 200 pg, 225 pg, 250 pg, or 275 pg to 300 pg of glucagon or a glucagon analogue, and in some cases, 150 ± 50 pg. The second dose can be administered during exercise or activity lasting more than 30, 40, 50, 60, 70, 80, 90, 100, 110, 120, 130, 140, 150, 160, 170, 180, 190, 200, 250, or 300 to 400 minutes or more. The second dose can be administered at 10, 15, 20, 25, 30, 40, 50, 60, 70, 80, 90, 100, 110, 120, 130, 140, 150, 160, 170, 180, 190, 200, 250, or 300 to 400 minutes or more after exercise or activity has begun.In other aspects, regardless of, or in conjunction with, the duration of exercise, the second dose may be administered when a certain intensity of exercise or activity has been reached. In one aspect, a second dose is administered after an exercise intensity of 60, 65, 70, 75, 80, or 85 percent of maximum heart rate or more, or after a MET of 1.5, 2, 2.5, 3, 3.5, 4, or more has been reached for 0.5, 1, 10, 20, 30, 40, 50, 60, or 70 minutes or more. In some aspects, 1, 2, 3, 4, or more doses may be administered during exercise. In some cases, a first, second, or third dose of glucagon or a glucagon analogue is administered after exercise. The dose may be 25 pg, 50 pg, 75 pg, 100 pg, 125 pg, 150 pg, 175 pg, 200 pg, 225 pg, 250 pg, or 275 pg to 300 pg of glucagon or a glucagon analogue, administered at 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 100, 110, 120, 130, 140, 150, 160, 170, 180, 190, or 200 to 250 minutes after exercise is completed. In some cases, glucagon or a glucagon analogue may be administered before, during, and after exercise. In certain aspects, 1, 2, 3, 4, or more doses can be administered during and / or after exercise. A variety of methods can be used to determine the intensity of exercise or activity. The term exercise or activity intensity refers to the amount of exercise or activity performed at a given level, such as the number of repetitions of an exercise or the duration of the exercise or activity. One method for determining exercise or activity intensity uses percentages of the exerciser's maximum heart rate. The exerciser's maximum heart rate can be calculated based on the exerciser's age, such as by subtracting the exerciser's age from 220. The exerciser then exercises at various intensity levels in such a way that their heart rate equals certain percentages (for example, between 60 and 85 percent) of their maximum heart rate.In another example, exercise intensity can be represented by the exercise intensity based on the Karvonen formula or as it is represented by exercise intensity as measured by metabolic equivalents (METs), which are a measure of oxygen consumption, and a Borg scale (subjective exercise intensity). Exercise intensity (METs) is a scale that represents the oxygen consumption in the body during or immediately after exercise caused by training or similar activities. More specifically, exercise intensity (METs) represents how much greater the current oxygen consumption is compared to the oxygen consumption measured in a resting condition (METs = 1) (the condition in which a user remains still in a chair for several minutes, the condition in which the user is in good physical health, and the condition in which the user feels relaxed).For example, in the case where the current oxygen consumption is twice the oxygen consumption measured in a resting condition, 2 is obtained as the exercise intensity (METs). In the methods of the present invention, an appropriate dosage of glucagon or a glucagon analogue can be administered. The administered dosage will, of course, vary depending on known factors, such as the pharmacodynamic characteristics of the particular compound, salt, or combination; the age, health, or weight of the subject; the nature and extent of the symptoms; the metabolic characteristics of the drug and the patient; the class of concurrent treatment; the frequency of treatment; or the desired effect. In certain respects, hypoglycemia can be treated by means of administering a formulation as described herein, which comprises an effective amount of glucagon. Glucagon formulations, such as the stable formulations described herein, can be used for parenteral injection of glucagon or a glucagon analogue. In some respects, one formulation is provided as an injectable formulation. The injectable formulation can be administered into the epidermal, dermal, or subcutaneous layer of an animal or human. In some respects, the formulations are administered intracutaneously. In some respects, glucagon formulations can be administered intravenously, intranasally, transdermally, or by inhalation. In some respects, a glucagon analogue is a glucagon peptidomimetic. Certain formulations utilize a glucagon or glucagon analogue formulation comprising glucagon or a glucagon analogue at a concentration of at least, and at most, approximately 0.1 mg, 1 mg, 10 mg, 50 mg, or 100 mg per mL to 150 mg, 200 mg, 300 mg, 400 mg, or 500 mg per mL, or up to the solubility limit of glucagon or a glucagon analogue. In certain aspects, the glucagon and the glucagon analogue are in a polar aprotic solvent system. In certain aspects, the polar aprotic solvent systems comprise a concentration of at least one ionization-stabilizing excipient that provides physical and chemical stability to the therapeutic agent. Certain formulations may comprise an ionization-stabilizing excipient at a concentration of at least, at most, or approximately 0.01 mM, 0.1 mM, 0.5 mM, 1 mM, 10 mM, or 50 mM to 10 mM, 50 mM, 75 mM, 100 mM, 500 mM, or 1000 mM, or up to the solubility limit of the ionization-stabilizing excipient in the polar aprotic solvent system. In certain aspects, the concentration of the ionization-stabilizing excipient is between 0.1 mM and 100 mM. The ionization-stabilizing excipient may be a suitable mineral acid, such as hydrochloric acid or sulfuric acid. In certain respects, the ionization-stabilizing excipient may be an organic acid, such as an amino acid, an amino acid derivative, or the salt of an amino acid or an amino acid derivative (examples include glycine, trimethylglycine (betaine), glycine hydrochloride, and trimethylglycine (betaine) hydrochloride). In another respect, the amino acid may be glycine or the amino acid derivative trimethylglycine. In other respects, the aprotic solvent system comprises DMSO.The aprotic solvent can be deoxygenated, for example, deoxygenated DMSO. In certain formulations, some formulations can be prepared by first adding an ionization-stabilizing excipient to a polar aprotic solvent system, followed by the addition of glucagon or a glucagon analogue. Alternatively, glucagon or a glucagon analogue can be initially solubilized in polar aprotic solvent systems, followed by the addition of the ionization-stabilizing excipient. In a further aspect, the ionization-stabilizing excipient and glucagon or a glucagon analogue can be solubilized simultaneously in the polar aprotic solvent system. In some formulations, the therapeutic agent is glucagon or a salt thereof. Other formulations can be prepared using other methods known in the art. Deímicjones The term glucagon refers to the glucagon peptide, its analogues, and the salt forms of any of them. The term "optimal stability and solubility" for a peptide refers to the pH environment in which the peptide's solubility is high (at or near its maximum relative to the pH profile, or suitable for product requirements), and its degradation is minimized compared to other pH environments. Notably, a peptide can have more than one pH of optimum stability and solubility. A person with ordinary experience in maternal nutrition can easily determine the optimum stability and solubility of a given peptide by consulting the literature or conducting tests. The term dissolution, as used herein, refers to a process by which a material or materials in a gaseous, solid, or liquid state become a solute or solutes, a dissolved component or components, of a solvent, forming a solution of the gas, liquid, or solid in the solvent. In certain respects, a therapeutic agent (e.g., glucagon or a glucagon analogue) or an excipient, e.g., an ionization-stabilizing excipient, is present up to its solubility limit or is completely solubilized. The term dissolve refers to a gas, liquid, or solid being incorporated into a solvent in order to form a solution. The term excipient, as used herein, refers to a natural or synthetic substance formulated together with the active agent or therapeutic ingredient (an ingredient other than the active ingredient) of a drug, which is included for the purpose of stabilization, bulking, or to confer an improvement to the therapeutic product over the active ingredient in the final dosage form, such as to facilitate drug absorption, reduce viscosity, improve solubility, adjust tonicity, alleviate discomfort at the injection site, depress freezing point, or improve stability.Excipients can also be useful in the manufacturing process, to help in the handling of the active substance concerned, such as by facilitating the flow or non-stick properties of the powder, as well as helping with in vitro stability such as preventing denaturation or agglomeration during the expected shelf life. The term therapeutic agent encompasses proteins, peptides, and pharmaceutically acceptable salts thereof. Useful salts are known to persons skilled in this field and include salts with inorganic acids, organic acids, inorganic bases, or organic bases. The useful therapeutic agents in the present invention are proteins and / or peptides that have a beneficial and often desired pharmacological effect after administration to a human or animal, either alone or in combination with other pharmaceutical excipients or inert ingredients.The terms peptide and peptide compound refer to polymers of amino acids or amino acid-like compounds (peptidomimetics) of up to approximately 200 amino acid residues linked together by amide (CONH) or other bonds. In some respects, a peptide may be as small as 150, 100, 80, 60, 40, 20, or 10 amino acids. Protein and protein compound refer to polymers of more than 200 amino acid residues linked together by amide bonds. Pharmaceutically acceptable analogues, derivatives, agonists, antagonists, and salts of any of the peptide or protein compounds disclosed herein are included under these terms.The terms also include peptides, proteins, peptide compounds, and protein compounds that have D amino acids, or modified, derived, or naturally occurring amino acids in the D or L configuration, and / or peptidomimetic units, as part of their structure. Analogue, when referring to a peptide or protein, means a modified peptide or protein in which one or more amino acid residues of the peptide or protein have been substituted by other amino acid residues, or in which one or more amino acid residues have been deleted from the peptide or protein, or in which one or more amino acid residues have been added to the peptide or protein, or any combination of these modifications. Such addition, deletion, or substitution of amino acid residues may occur at any point, or at multiple points, along the primary structure comprising the peptide or protein, including at the N-terminus and / or the C-terminus of the peptide or protein. Analogue also includes functional analogues or mimetics / peptidomimetics. Derivative, in relation to a parent peptide or protein, refers to a chemically modified parent peptide or protein, or an analogue thereof, wherein at least one substituent is not present in the parent peptide or protein, or an analogue thereof. A non-limiting example is a parent peptide or protein that has been covalently modified. Typical modifications include amides, carbohydrates, polysaccharides, glycans, alkyl groups, acyl groups, esters, pegylations, and the like. A single-phase solution refers to a solution prepared from a therapeutic agent dissolved in a solvent, or in solvent systems (e.g., a mixture of two or more solvents), where the therapeutic agent is completely dissolved in the solvent and no particulate matter is visible, such that the solution can be described as optically transparent. A single-phase solution may also be referred to as a single-phase system, and is distinguished from a two-phase system in that the latter consists of particulate matter (e.g., powder) suspended in a fluid. Inhibit, reduce, mitigate, or any variation of these terms, include any measurable decrease or complete inhibition to achieve a desired result. Mitigation or any variation of these terms includes any improvement or benefit to a subject with respect to an intended condition. Effective, or treating, or preventing, or any variation of these terms, mean that they are suitable for carrying out a desired, expected, or intended result. Chemical stability, when referring to a therapeutic agent, refers to an acceptable percentage of degradation products produced by chemical pathways, such as oxidation, hydrolysis, fragmentation, and / or other chemical degradation pathways. Specifically, a formulation is considered chemically stable if no more than approximately 20 percent of the decomposition products form after one year of storage at the product's intended storage temperature (e.g., room temperature); or after one year of storage at 25°C and 60 percent relative humidity; or after one month, and preferably three months, of storage at 40°C and 75 percent relative humidity.In some embodiments, a chemically stable formulation has less than 20 percent, less than 15 percent, less than 10 percent, less than 5 percent, less than 4 percent, less than 3 percent, less than 2 percent, or less than 1 percent of the decomposition products formed after a prolonged period of storage at the intended storage temperature of the product. Physical stability, when referring to a therapeutic agent, refers to an acceptable percentage of aggregates (e.g., dimers, trimers, and larger forms) that form. In particular, a formulation is considered physically stable if no more than approximately 15 percent aggregates form after one year of storage at the product's intended storage temperature (e.g., room temperature); or after storage at 25°C at 60 percent relative humidity for one year; or after storage at 40°C at 75 percent relative humidity for one month, and preferably three months.In some forms, a physically stable formulation has less than 15 percent, less than 10 percent, less than 5 percent, less than 4 percent, less than 3 percent, less than 2 percent, or less than 1 percent of aggregates formed after a prolonged period of storage at the product's intended storage temperature. A stable formulation refers to a formulation in which at least approximately 65 percent of the therapeutic agents (e.g., peptides or their salts) remain chemically and physically stable after two months of storage at room temperature. Particularly preferred formulations are those in which at least approximately 80 percent, 85 percent, 90 percent, 91 percent, 92 percent, 93 percent, 94 percent, 95 percent, 96 percent, 97 percent, 98 percent, or 99 percent of the therapeutic agent remains chemically and physically stable under these storage conditions. Especially preferred stable formulations are those that do not exhibit degradation following sterilizing irradiation (e.g., gamma, beta, or electron beam). As used herein, parenteral administration refers to the administration of a therapeutic agent to a patient by means of a route other than the alimentary canal - any administration other than by means of the digestive tract. As used herein, parenteral injection refers to the administration of therapeutic agents (e.g., peptides or small molecules) by means of an injection under or through one or more layers of the skin or mucous membranes of an animal, such as a human. Standard parenteral injections are administered into the subcutaneous, intramuscular, or intradermal region of an animal or subject, such as a human. These deeper locations are targeted because the tissue expands more readily relative to more superficial dermal sites to accommodate the injection volumes required to deliver most therapeutic agents, e.g., 0.1 to 3.0 cc (mL). The term intracutaneous encompasses administration into the epidermal, dermal, or subcutaneous layer of the skin. As used herein, the term polar aprotic solvent refers to a polar solvent that does not contain acidic hydrogen and therefore does not act as a hydrogen bond donor. Polar aprotic solvents include, but are not limited to, DMSO, DMF, ethyl acetate, n-methyl-pyrrolidone (NMR), dimethylacetamide (DMA), and propylene carbonate. As used herein, the term polar aprotic solvent system refers to a solution in which the solvent is either a single polar aprotic solvent (e.g., pure DMSO), or a mixture of two or more polar aprotic solvents (e.g., a mixture of DMSO and NMP). As used herein, residual moisture may refer to the residual moisture in the drug powder immediately after preparation by the manufacturer / supplier. Typical powders often have a residual moisture content of up to 10 percent (w / w). When these powders are dissolved in a polar aprotic solvent system, the residual moisture in the powder is incorporated into the formulation. Additionally, polar aprotic solvents may also contain some residual moisture. For example, a freshly opened bottle of USP-grade DMSO typically contains up to 0.1 percent (w / w) moisture. Residual moisture is distinct from added moisture, where water is intentionally added to the formulation, for example, to serve as a co-solvent or to depress the freezing point of the polar aprotic solvent system.Moisture can also be introduced into the formulation during the addition of an ionization-stabilizing excipient (e.g., through the addition of a mineral acid from an aqueous supply solution (e.g., NHCl or H₂SO₄)). The total moisture content (% wt / wt, unless otherwise reported) in a formulation immediately after preparation is due to contributions from both residual moisture and added moisture. The terms "around" or "approximately" or "substantially unchanged" are defined as being close to what is understood by a person with ordinary experience in this field, and in a non-limiting manner, the terms are defined as within 10 percent, preferably within 5 percent, more preferably within 1 percent, and most preferably within 0.5 percent. In addition, "substantially non-aqueous" refers to less than 5 percent, 4 percent, 3 percent, 2 percent, 1 percent, or less by weight or volume of water. A "pharmaceutically acceptable" ingredient, excipient, or component is one that is suitable for use with humans and / or animals without undue adverse side effects (such as toxicity, irritation, and allergic response) occurring in a manner commensurate with a reasonable benefit / risk ratio. Pharmaceutically acceptable vehicle means a solvent, suspending agent, or pharmaceutically acceptable vehicle for delivering a drug compound of the present invention to a mammal, such as a human being. As used herein, an ionization-stabilizing excipient is an excipient that stabilizes and / or maintains a particular ionization state for a therapeutic agent. In certain respects, the ionization-stabilizing excipient may be, or may include, a molecule that donates at least one proton under appropriate conditions or that is a source of protons. According to the Brønsted-Lowry definition, an acid is a molecule that can donate a proton to another molecule, which, by accepting the donated proton, can therefore be classified as a base. As used in this application, and as will be understood by a person of expertise, the term “proton” refers to the hydrogen ion, the hydrogen cation, or H+. The hydrogen ion has no electrons and consists of a nucleus that usually consists exclusively of a proton (for the most common isotope of hydrogen, protium).Specifically, a molecule that can donate at least one proton to a therapeutic agent is considered to be a source of acid or protons, regardless of whether it is fully ionized, mostly ionized, partially ionized, mostly not ionized, or not ionized at all in the polar aprotic solvent. As used herein, a mineral acid is an acid derived from one or more inorganic compounds. Accordingly, mineral acids may also be referred to as inorganic acids. Mineral acids can be monoprotic or polyprotic (e.g., diprotic, triprotic, etc.). Examples of mineral acids include hydrochloric acid (HCl), sulfuric acid (H₂SO₄), and phosphoric acid (H₃PO₄). As used herein, an organic acid is an organic compound with acidic properties (i.e., it can function as a proton source). Carboxylic acids are an example of organic acids. Other well-known examples of organic acids include, but are not limited to, alcohols, thiols, enols, phenols, and sulfonic acids. Organic acids can be monoprotic or polyprotic (e.g., diprotic, triprotic, etc.). “Charge profile”, charge state, ionization, ionization state, and ionization profile can be used interchangeably and refer to the ionization state due to protonation and / or deprotonation of the ionogenic groups of the peptide. As used herein, a co-formulation is a formulation containing two or more therapeutic agents dissolved in a polar aprotic solvent system. The therapeutic agents may belong to the same class, or they may belong to different classes. An amphoteric species is a molecule or ion that can react as both an acid and a base. These species can either donate or accept a proton. Examples include amino acids, which possess both amine and carboxylic acid functional groups. Amphoteric species also include amphiprotic molecules, which contain at least one hydrogen atom and have the ability to donate or accept a proton. The use of the words "a" or "one" when used in conjunction with the term comprising in the claims and / or in the descriptive memorandum may mean one, but is also consistent with the meaning of one or more, at least one, and one or more than one. The words that comprise (and any form of comprise, such as comprise and comprise), that have (and any form of have, such as have and has), that include (and any form of include, such as include and includes) or that contain (and any form of contain, such as contains and contain) are inclusive or have no fixed limits and do not exclude additional elements or steps of the method not mentioned. Other objects, features, and advantages of the present invention will become apparent from the following detailed description. It should be understood, however, that the detailed description and examples, although indicating specific embodiments of the invention, are given by way of illustration only. Furthermore, it is anticipated that changes and modifications within the spirit and scope of the invention will become apparent to persons skilled in the field from this detailed description. The compositions and methods for making and using them of the present invention may comprise, consist essentially of, or consist of the particular ingredients, components, mixtures, steps, methods, etc. that are disclosed throughout the description. Other embodiments of the invention are discussed throughout this application. Any embodiment discussed with respect to one aspect of the invention also applies to other aspects of the invention, and vice versa. Each embodiment as described herein is understood to be an embodiment of the invention applicable to all aspects of the invention. It is contemplated that any embodiment discussed herein may be implemented with respect to any method or composition of the invention, and vice versa. Brief Description of Drawings The following drawings form part of this specification and are included to further demonstrate certain aspects of the present invention. The invention may be better understood by reference to one or more of these drawings in combination with the detailed description of the features presented herein. Figures 1A 1B. Diagram of a representative study protocol. Figure 2. YSI Glucose during exercise / early recovery and after food. Figure 3. Hypoglycemia and hyperglycemia during exercise / early recovery. Figure 4. Glucagon during exercise / early recovery and after food. Figure 5. Insulin during exercise / early recovery. Figure 6. Hypoglycemia and hyperglycemia after food. Detailed Description of the Invention Different modalities utilize glucagon or glucagon analog formulations. These formulations may include, but are not limited to, stable glucagon or glucagon analog formulations. Examples of such stable glucagon or glucagon analog formulations are described herein. In certain aspects, the present invention provides methods for treating exercise-associated or exercise-induced hypoglycemia by administering to a subject a therapeutic agent in an amount effective for treating, mitigating, or preventing the condition. In some embodiments, a therapeutic method of the present invention comprises treating hypoglycemia by administering to a subject who has hypoglycemia or who is at risk of developing hypoglycemia a therapeutic agent for hypoglycemia in a stable formulation in an amount effective for treating hypoglycemia. In some embodiments, the subject is administered a formulation comprising glucagon or a glucagon analogue.In some respects, hypoglycemia can be caused by diseases, conditions, and disorders related to diabetes or not related to diabetes that affect glycemic regulation when a subject exercises. A. Treatment of exercise-associated hypoglycemia In certain aspects, the present invention provides methods for the treatment of exercise-associated or exercise-induced hypoglycemia by means of administering to a subject a therapeutic agent in an amount effective for the treatment, mitigation, or prevention of the condition. Certain modalities refer to methods for preventing or mitigating exercise-associated hypoglycemia through the use of glucagon or a glucagon analogue, and specifically to a mini-dose of glucagon or a glucagon analogue. In certain aspects, a mini-dose is between 25 mcg, 50 mcg, 75 mcg, 100 mcg, 125 mcg, 150 mcg, and 175 mcg, 200 mcg, 225 mcg, 250 mcg, 275 mcg, and 300 mcg, which include all values ​​and intervals between them. In certain aspects, the mini-dose of glucagon is from 125 mcg to 175 mcg, which includes 150 mcg. In certain aspects, glucagon is administered subcutaneously just before exercise in patients with TID. In a certain respect, just before exercise includes 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, or 80 to 85 minutes before the start of exercise.The onset of exercise may include the beginning of activity that results in an increase in the subject's heart rate of more than 10, 20, 30, 40, or 50 percent above the resting heart rate. A pre-exercise mini-dose of glucagon may produce greater glucose stability than reductions in basal insulin or extra carbohydrate intake. In some cases, a mini-dose of glucagon may be administered to mitigate post-exercise nocturnal blood glucose levels. The method may further include monitoring or continuous monitoring of blood glucose. In some cases, an initial or subsequent dose of glucagon or a glucagon analogue may be administered if a subject's insulin or blood glucose levels fall below a threshold level. In some cases, the threshold level is 80 mg, 70 mg, 60 mg, or 50 mg / dL. As described by the Workgroup of the American Diabetes Association and the Endocrine Society (Seaquist et al. (2013), Diabetes Care, Vol 36, pp. 1384-1395) with respect to hypoglycemia, a single threshold value for plasma glucose concentration is not usually assigned to define hypoglycemia in diabetes because glycemic thresholds for the symptoms of hypoglycemia (among other responses) change in order to reduce plasma glucose concentrations after recent antecedent hypoglycemia and up to the highest plasma glucose concentrations in patients with poorly controlled diabetes and infrequent hypoglycemia. However, an alert threshold can be defined to direct the attention of both patients and caregivers to the potential harm associated with hypoglycemia. Patients at risk of hypoglycemia (i.e., those treated with a sulfonylurea, glinide, or insulin) should be alert to the possibility of developing hypoglycemia at a self-monitored plasma glucose concentration—or a subcutaneous glucose concentration from continuous glucose monitoring—of <70 mg / dL (<3.9 millimoles per liter). Because this is higher than the glycemic threshold for symptoms in both nondiabetic individuals and those with well-controlled diabetes, it generally provides time to prevent a clinical hypoglycemic episode and allows some leeway for the limited accuracy of monitoring devices at low glucose levels. Severe hypoglycemia is an event that requires assistance from another person to actively administer carbohydrates, glucagon, or take other corrective measures. Plasma glucose concentrations may not be available during an event, but neurological recovery following the return of plasma glucose to normal is considered sufficient evidence that the event was induced by low plasma glucose. Typically, these events begin to occur at plasma glucose concentrations of <50 mg / dL (2.8 mM / L). Documented symptomatic hypoglycemia is an event during which typical symptoms of hypoglycemia are accompanied by a measured plasma glucose concentration of <70 mg / dL (<3.9 mM / L).Asymptomatic hypoglycemia is an event that is not accompanied by the typical symptoms of hypoglycemia but with a measured plasma glucose concentration < 70 mg / dL (< 3.9 millimoles per liter). Probable symptomatic hypoglycemia is an event during which the typical symptoms of hypoglycemia are not accompanied by a plasma glucose determination but were presumably caused by a plasma glucose concentration < 70 mg / dL (< 3.9 millimoles per liter). Pseudo-hypoglycemia is an event during which the person with diabetes reports any of the typical symptoms of hypoglycemia with a measured plasma glucose concentration > 70 mg / dL (> 3.9 millimoles per liter) but approaching that level. The formulations of the present invention can also be used for the treatment of hyperinsulinemic hypoglycemia, which broadly refers to the condition and effects of low blood glucose levels caused by an excess of insulin. Hyperinsulinemic hypoglycemia can occur due to endogenous insulin, for example, in exercise-induced hypoglycemia. The most common type of severe, but usually transient, hyperinsulinemic hypoglycemia arises from the administration of exogenous insulin in patients with type 1 diabetes. This type of hypoglycemia can be defined as iatrogenic hypoglycemia and is a limiting factor in the glycemic management of type 1 and type 2 diabetes. Nocturnal hypoglycemia (nighttime hypoglycemia) is a common type of iatrogenic hypoglycemia that occurs in patients taking exogenous insulin. In some embodiments, a therapeutic method of the present invention comprises treating a subject with diabetes before, during, after, before and during, before and after, or before, during, and after exercise by administering to the subject with diabetes a therapeutic agent in a stable formulation in an effective amount to stabilize or increase glucose levels or glucagon / insulin ratios during and / or after exercise. In some embodiments, the subject is administered a stable formulation comprising glucagon or a glucagon analogue. Determining an effective quantity or dose is well within the capabilities of those skilled in this field, particularly given the detailed description provided herein. Generally speaking, formulations for delivering these doses may contain one or more peptides, or peptide analogues (collectively referred to as peptide, unless expressly excluded), where each peptide is present at a concentration of approximately 0.1 mg / mL up to the peptide's solubility limit in the formulation. This concentration is preferably from approximately 1 mg / mL to approximately 100 mg / mL.In certain aspects the concentration is approximately 1 mg per mL, approximately 5 mg per mL, approximately 10 mg per mL, approximately 15 mg per mL, approximately 20 mg per mL, approximately 25 mg per mL, approximately 30 mg per mL, approximately 35 mg per mL, approximately 40 mg per mL, approximately 45 mg per mL, approximately 50 mg per mL, approximately 55 mg per mL, approximately 60 mg per mL, approximately 65 mg per mL, approximately 70 mg per mL, approximately 75 mg per mL, approximately 80 mg per mL, approximately 85 mg per mL, approximately 90 mg per mL, approximately 95 mg per mL, or approximately 100 mg per mL. The formulations of the present invention may be for subcutaneous, intradermal, or intramuscular administration (e.g., by injection or infusion). In some embodiments, the formulation is administered subcutaneously. Glucagon or glucagon analogue formulations can be administered by infusion or injection using any suitable device. For example, a glucagon or glucagon analogue formulation can be placed in a syringe (e.g., a pre-filled syringe), a pencil injection device, an auto-injector, or a pump device. In some modalities, the injection device is a multi-dose injection pump or a multi-dose auto-injector. The glucagon or glucagon analogue formulation can be presented in the device in such a form that it readily flows out of the needle upon activation of an injection device, such as an auto-injector, to deliver the peptide drugs.Suitable pen injection devices or auto-injectors include, but are not limited to, pen injection devices or auto-injectors manufactured by Becton-Dickenson, Swedish Healthcare Limited (SHL Group), YpsoMed AG, and similar manufacturers. Suitable pump devices include, but are not limited to, pump devices manufactured by Tandem Diabetes Care, Inc., Deisys Pharmaceuticals, and similar manufacturers. In some forms, glucagon or glucagon analogue formulations are provided ready for administration in a bottle, cartridge, or pre-filled syringe. The formulations can be administered via oral, parenteral (e.g., intramuscular, intraperitoneal, intravenous or intra-arterial, ICV, intrasystemic, subcutaneous injection, or implant), inhalation, nasal, vaginal, rectal, sublingual and / or topical (e.g., transdermal or local) routes, and can be formulated alone or together in a suitable dosage unit with a pharmaceutically acceptable vehicle, carrier, diluent or excipient or a mixture thereof, appropriate for each route of administration. In certain respects, glucagon or glucagon analogue formulations can be administered parenterally by injection, infusion, or implantation for local or systemic administration. Parenteral administration, as used herein, includes intravenous, intra-arterial, intraperitoneal, intrathecal, intraventricular, intraurethral, ​​intrasternal, intracranial, intramuscular, intrasynovial, and subcutaneous administration. Additionally, glucagon or glucagon analogue formulations can be administered intranasally or by inhalation into the respiratory tract. Pharmaceutical compositions can be provided as an aerosol or solution for delivery via a pressurized container, pump, spray, atomizer (such as an electrohydrodynamic atomizer that produces a fine mist), or nebulizer, alone or in combination with a suitable propellant, such as 1,1,1,2-tetrafluoroethane or 1,1,1,2,3,3,3-heptafluoropropane. In some formulations, a stable formulation is used to formulate a drug for the treatment of exercise-associated hypoglycemia. In some formulations, the stable formulation comprises glucagon, a glucagon analogue (e.g., dasiglucagon), or a salt thereof (e.g., glucagon acetate). In some formulations, the stable formulation is used to formulate a drug for the treatment of diabetic patients before, during, or after exercise. Certain aspects of the methods described herein have been implemented using a pump-based system. Pump-based systems used for administering glucagon or glucagon analogue compositions may include closed-loop, open-loop, or non-loop systems. Glucagon or glucagon analogue formulations that can be used with these systems are designed to be contained or stored in a pump container without the need for reconstitution (i.e., they are readily available for administration to the patient / subject from the pump container).Furthermore, glucagon or glucagon analogue formulations can be stable at non-refrigerated temperatures (20°C to 35°C) for extended periods (> 2 months) (i.e., formulations can be safely stored in the pump container without risking a substantial loss in glucagon activity in the formulation or without risking the formation of insoluble clumps that will inhibit delivery and clog the infusion apparatus). The pump-based system may include: (1) a glucose sensor that is inserted or can be inserted into a patient and is capable of measuring blood glucose levels (e.g., either directly by means of contact with the patient's blood or indirectly by means of contact with the patient's interstitial fluid); (2) a transmitter that sends glucose information from the sensor to a monitor (e.g., by means of radio frequency transmission); (3) a pump that is designed in such a way as to store and deliver glucose or the glucagon formulation or a glucagon analogue to the patient; and / or (4) a monitor (e.g., one that can be integrated into the pump device or a separate monitor) that displays or records glucose levels.In a closed-loop system, the glucose monitor may be able to adjust the delivery of the glucagon or glucagon analog formulation to the patient via the pump, based on an algorithm. This closed-loop system requires little to no patient intervention and instead actively monitors blood glucose levels and delivers the necessary amount of the glucagon or glucagon analog formulation to the patient to maintain an appropriate glucose level and prevent hypoglycemia. In an open-loop system, the patient would actively participate by reading their glucose monitor and adjusting the delivery rate or dose based on the information provided by the monitor. In a non-loop system, the pump would deliver the glucagon or glucagon analog formulation at a fixed (or basal) dose.The loop-free system can be used without a glucose monitor and without a glucose sensor if desired. In certain respects, a glucagon or glucagon analogue delivery apparatus may include a reservoir containing a composition comprising glucagon, a glucagon analogue, or a salt form thereof; a sensor configured to measure a patient's blood glucose level; and an electronic pump configured to deliver at least a portion of the composition to a patient via the intradermal, subcutaneous, or intramuscular route, based on the patient's measured blood glucose level. The sensor may be positioned on the patient in such a manner as to make contact with the patient's blood, the patient's interstitial fluid, or both.The sensor can be configured to transmit data (e.g., wirelessly, via radio frequency, or via a wired connection) to a processor configured to control the operation of the electronic pump. The processor can be configured to control the pump's operation based, at least in part, on the data obtained by the sensor.In one instance, the processor can be configured in such a way as to control the operation of the pump in order to inject intradermally, subcutaneously, or intramuscularly at least a portion of the composition or formulation if the data obtained by means of the sensor indicates a glucose level below a defined threshold, or to give an indication that a defined threshold will be violated within a particular time period (e.g., an indication of impending hypoglycemia or an indication that blood glucose levels will fall to below 70 mg, 60 mg, or 50 mg per dL within a certain time period (e.g., within 30, 25, 20, 15, 10, 9, 8, 7, 6, 5, 4, 3, 2, or 1 minutes).This indication can be determined by identifying a downward trend in blood glucose levels (for example, using a blood glucose monitoring device), as well as by the rate or trajectory of this downward trend. The glucagon delivery device may also include a monitor configured to communicate information indicating the patient's glucose level. The monitor may include a speaker, a display, or both. The monitor may be configured to issue an alert when the patient's glucose level is estimated to be at a defined threshold.Furthermore, the device can be configured in such a way as to allow manual adjustment of at least one delivery rate and dose of the composition delivered intradermally, subcutaneously, or intramuscularly by means of the pump. B. Therapeutic agents The therapeutic agents in the context of the present invention encompass peptide or protein compounds, small-molecule drugs, and pharmaceutically acceptable salts thereof. In certain respects, when the therapeutic agent is present in the deoxygenated polar aprotic solvent, the stability of the therapeutic agent can be further improved compared to the same therapeutic agent present in an untreated polar aprotic solvent. The increased stability can be attributed, at least in part, to a reduction in the oxidative degradation of the therapeutic agent or the oxidative degradation of the polar aprotic solvent, or both.A skilled person is aware of which therapeutic agent is appropriate for the treatment of certain diseases or conditions and would be able to administer effective amounts of a therapeutic agent in a formulation as described herein, for the treatment of a disease or condition. ocaonn / nznz / E / Y Non-limiting examples of peptides and proteins (and salts thereof) that may be used in the context of the present invention include, but are not limited to, giucagon or analogues thereof. The therapeutic agent of the invention can be administered intracutaneously for the prevention, diagnosis, relief, treatment, or cure of disease. Examples of proteins and protein compounds that can be formulated and used in the delivery system according to the present invention include those proteins that have biological activity or that can be used for the treatment of a disease or other pathological conditions. Any suitable dosage of the peptide or peptides may be formulated. Generally, the peptide (or, in some formulations comprising two or more peptides, each of the peptides) is present in the formulation in an amount ranging from approximately 0.1 mg per mL to approximately 100 mg per mL. In some formulations, the peptide is present in the formulation in an amount ranging from approximately 5 mg per mL to approximately 60 mg per mL. In other formulations, the peptide is present in the formulation in an amount ranging from approximately 10 mg per mL to approximately 50 mg per mL. In still other formulations, the peptide is present in the formulation in an amount ranging from approximately 1 mg per mL to approximately 15 mg per mL.In still other formulations, the peptide is present in the formulation in an amount ranging from approximately 0.5 mg per mL to approximately 5 mg per mL. In still other formulations, the peptide is present in the formulation in an amount ranging from approximately 1 mg per mL to approximately 50 mg per mL. In some embodiments, the formulations may also include an antioxidant. In other embodiments, the formulations may also include a chelating agent. In still other embodiments, the formulations may also include a preservative. C. Formulations The formulations that can be used in the methods described herein may include a therapeutic agent present in a polar aprotic solvent system. In some respects, polar aprotic solvent systems may contain at least one ionization-stabilizing excipient. The therapeutic agent may be dissolved (e.g., completely or partially solubilized) or suspended (completely or partially) in the polar aprotic solvent system. Furthermore, a formulation may be structured as a single-phase solution, a paste or aqueous paste, a gel, an emulsion, or a suspension. In some formulations, the therapeutic agent is present in a pure polar aprotic solvent, meaning it does not contain a co-solvent. In other formulations, the therapeutic agent is present in a solvent system that is a mixture of two or more polar aprotic solvents (i.e., a polar aprotic solvent system). An example would be a 75 / 25 (v / v) mixture of DMSO and NMR. In some formulations, however, a co-solvent may be used, where a co-solvent is mixed into one or more polar aprotic solvents. Non-limiting examples of co-solvents include water, ethanol, propylene glycol (PG), glycerol, and mixtures thereof. In certain aspects, water as a co-solvent may be specifically excluded or limited; that is, the co-solvent may be a non-aqueous co-solvent.The co-solvent may be present in the formulation in an amount that is in the range of approximately 0.5 percent (weight / volume) to approximately 50 percent (weight / volume), for example, approximately 1 percent, approximately 5 percent, approximately 10 percent, approximately 15 percent, approximately 20 percent, approximately 25 percent, approximately 30 percent, approximately 35 percent, or approximately 40 percent (weight / volume).In some embodiments, the co-solvent is present in the formulation in an amount that is in the range of approximately 10 percent (w / v) to approximately 50 percent (w / v), approximately 10 percent (w / v) to approximately 40 percent (w / v), approximately 10 percent (w / v) to approximately 30 percent (w / v), approximately 10 percent (w / v) to approximately 25 percent (w / v), approximately 15 percent (w / v) to approximately 50 percent (w / v), approximately 15 percent (w / v) to approximately 40 percent (w / v), approximately 15 percent (w / v) to approximately 30 percent (w / v), or approximately 15 percent (w / v) to approximately 25 percent. (weight / volume). Furthermore, a glucagon or glucagon analogue formulation may include one or more excipients. In some formulations, the excipient is selected from sugars, starches, sugar alcohols, antioxidants, chelating agents, and preservatives. Examples of suitable sugar excipients include, but are not limited to, trehoiso, glucose, sucrose, etc. Examples of suitable starches for excipient stabilization include, but are not limited to, hydroxyethyl starch (HES). Examples of suitable sugar alcohols (also referred to as polyols) for excipient stabilization include, but are not limited to, mannitol and sorbitol. Examples of suitable antioxidants include, but are not limited to, ascorbic acid, cysteine, methionine, monothioglycosides, sodium thiosulfate, sulfites, BHT, BHA, ascorbyl palmitate, propyl gallate,N-acetyl-L-cysteine ​​(NAC) and vitamin E. Examples of suitable chelating agents include, but are not limited to, EDTA, disodium EDTA (edetate disodium), tartaric acid and its salts, glycerin, and citric acid and its salts. Examples of suitable inorganic salts include sodium chloride, potassium chloride, calcium chloride, magnesium chloride, calcium sulfate, and magnesium sulfate. Examples of suitable preservatives include, but are not limited to, benzyl alcohols, methylparabens, propylparabens, and mixtures thereof. Additional formulation components include local anesthetics such as lidocaine or procaine. In some formulations, an additional stabilizing excipient is present in the formulation in an amount that is in the range of approximately 0.05 percent (weight / volume) to approximately 60 percent (weight / volume),from approximately 1 percent (weight / volume) to approximately 50 percent (weight / volume), from approximately 1 percent (weight / volume) to approximately 40 percent (weight / volume), from approximately 1 percent (weight / volume) to approximately 30 percent (weight / volume), from approximately 1 percent (weight / volume) to approximately 20 percent (weight / volume), from approximately 5 percent (weight / volume) to approximately 60 percent (weight / volume), from approximately 5 percent (weight / volume) to approximately 50 percent (weight / volume), from approximately 5 percent (weight / volume) to approximately 40 percent (weight / volume), from approximately 5 percent (weight / volume) to approximately 30 percent (weight / volume), from approximately 5 percent (weight / volume) to approximately the 20 percent (weight / volume),from approximately 10 percent (weight / volume) to approximately 60 percent (weight / volume), from approximately 10 percent (weight / volume) to approximately 50 percent (weight / volume), from approximately 10 percent (weight / volume) to approximately 40 percent (weight / volume), from approximately 10 percent (weight / volume) to approximately 30 percent (weight / volume), or from approximately 10 percent (weight / volume) to approximately 20 percent (weight / volume). In some embodiments, the additional stabilizing excipient is present in the formulation in an amount that is approximately, at most, or at least 0.1, 0.5, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55 or 60 percent (weight / volume). D. Kits / Containers Kits for use in certain aspects of the present invention are also contemplated. For example, a formulation of glucagon or a glucagon analogue may be included within a kit. A kit may include a container. In one aspect, for example, the formulation may be contained within a container that is ready for administration to a subject without the formulation needing to be reconstituted or diluted. That is, the formulation to be administered may be stored in the container and can be used easily as required. The container may be a device. The device may be a syringe (for example, a pre-filled syringe), a pen injection device, an auto-injector device, or a device that can pump or administer the formulation (for example, automatic or non-automatic external pumps, implantable pumps, etc.).or an infusion bag, pen devices or auto-injectors include, but are not limited to, pen devices or auto-injectors manufactured by Becton-Dickenson, Swedish Healthcare Limited (SHL Group), YpsoMed Ag, and similar companies. Suitable pump devices include, but are not limited to, pump devices manufactured by Tándem Diabetes Caro, Inc., Deisys Pharmaceuticais, and similar companies. ocaonn / nznz / E / Y EXAMPLES The following examples, as well as the figures, are included to demonstrate preferred embodiments of the invention. It should be appreciated by persons skilled in the art that the techniques disclosed in the examples or figures represent the techniques discovered by the inventors that work well in the practice of the invention and, therefore, may be considered to constitute the preferred modes of its practice. However, persons skilled in the art will appreciate that, with respect to the present invention, many changes can be made to the specific embodiments disclosed and the same or a similar result can still be obtained without departing from the spirit and scope of the invention. Example 1 Use of mini-doses of glucagon to prevent exercise-induced hypoglycemia in type 1 diabetes (T1D) An example of a study to evaluate the effectiveness of mini-dose glucagon in preventing or mitigating exercise-induced hypoglycemia in type 1 diabetes (T1D) may include a 4-period randomized crossover study. Visit Schedule. (1) Screening or Baseline Visit. This visit will be used to assess eligibility and may include VO2 max determination for fitness assessment and exercise intensity determination for the study. (2) Randomized Crossover Study. Each participant will undergo four aerobic exercise sessions (in a randomized order), with different glucose regulation strategies, including: (i) Control: Fasted exercise, no basal insulin reduction; (ii) Strategy 1: Fasted exercise, basal insulin reduction only (50 percent reduction in basal rate five minutes before exercise, for the duration of exercise); (iii) Strategy 2: Fasted exercise, no basal adjustment + glucose tablets before exercise (orally - 40 grams total); (iv) Strategy 3: Fasted exercise, no basal adjustment + mini-dose of glucagon before exercise.In all four sessions, aerobic exercise will be performed in a fasted state (before a standardized meal) for 45 minutes at approximately 50 to 55 percent of the participant's previously determined aerobic capacity. The selection strategies for the sessions will be blinded. The primary outcome for this study will be the glycemic response during exercise and shortly thereafter during recovery. Safety outcomes may include hypoglycemic events. A risk-based monitoring approach will be followed, consistent with the EDA “Guidance for Industry Oversight of Clinical Investigations - A Risk-Based Approach to Monitoring” (August 2013). Study Population. The crossover study will include participants who complete the study. Participants may be replaced if a participant does not complete the entire protocol. In order to be eligible for the study, a participant must meet the inclusion criteria and must not meet any of the exclusion criteria. Inclusion criteria may include: (1) Clinical diagnosis of presumed autoimmune type 1 diabetes, receiving daily insulin; (2) Age 18 to 65 years; (3) Duration of T1D > 2 years; (4) Random C-peptide < 0.6 ng / mL; (5) Use of CSH (e.g., insulin pump) for at least 6 months, with no plans to discontinue pump use during the study; (6) Regular exercise, i.e., 30 minutes of moderate-to-vigorous aerobic activity > 3 times per week; (7) Body mass index (BMI) < 30 kg / m².(8) Women must meet one of the following criteria: (a) with the potential to become pregnant and not currently pregnant or breastfeeding, and agrees to use an accepted contraceptive regimen as described in the study procedure manual throughout the entire duration of the study; or (b) without the potential to become pregnant, defined as a woman who has undergone a hysterectomy or tubal ligation, is considered clinically infertile, or is in a menopausal state (at least 1 year without menstruation); (9) In good general health without conditions that may influence the outcome of the study, and in the investigator's judgment, a good candidate for the study based on a review of available medical history, physical examination, and clinical laboratory evaluations; (10) Willing to adhere to the protocol requirements for the duration of the study;(11) You must enroll in the T1D exchange clinical registry or wish to join the registry.; Exclusion criteria may include: (1) one or more severe hypoglycemic episodes in the past 12 months (as defined by an episode requiring third-party assistance for treatment); (2) active diabetic retinopathy (PDR or VH in the past 6 months) that could potentially be worsened by the exercise protocol; (3) peripheral neuropathy with numb feet; (4) cardiovascular autonomic neuropathy with an inappropriate heart rate response to exercise (could be diagnosed by screening ECG: rule out tachycardia with a fixed RR interval); (5) use of antidiabetic medications other than insulin; (6) use of beta-blockers; (7) use of agents that affect hepatic glucose production, such as beta-adrenergic agonists or xanthine derivatives; (8) use of pramlintide; (9) currently following a very low-calorie or other weight-loss diet.(10) Participation in other studies involving administration of an investigational drug or device within 30 days or 5 half-lives, whichever is longer, prior to screening for the current study, or planning to participate in another such study during participation in the currently described study. Patient screening visit and enrollment. Participants will have a screening visit to assess eligibility through the submission of a medical history and a physical examination by a study investigator. Screening laboratory studies will be collected. A urine pregnancy test will be administered to women of childbearing potential. Clinical sites will maintain a record of the reasons why screened participants do not enroll. The following laboratory test results, which are routinely obtained as part of routine care, will be evaluated by the investigator as part of the overall assessment to determine eligibility. The time periods prior to enrollment during which each test must have been performed are given in parentheses: Basic Metabolic Panel (within 3 months); Complete Blood Count (within 3 months); Liver Function Panel (within 6 months); Random C-peptide (since diagnosis, subjects must have <0.6 ng / mL); Lipids (within 6 months); Thyroid-Stimulating Hormone (within 12 months); HbA1c will be measured using a point-of-care device or in a local laboratory if this is not available within 3 months prior to enrollment; Urine Pregnancy Test if indicated (see inclusion criteria). A participant history will be provided and extracted from available medical records regarding the participant's diabetes history, current diabetes management, other past and present medical conditions, past and present medications, drug allergies, and family history. The study investigator or their designee (an endocrinologist, endocrinologist resident, endocrine nurse practitioner, or physician assistant) will conduct a standard physical examination (including vital signs and measurements of height and weight). The study will be discussed with potential participants. A copy of the consent form will be provided to the participant, and another copy will be added to the participant's medical record. Written informed consent will be obtained before performing any study-specific procedures that are not part of the participant's routine care. As part of the informed consent process, each participant will be asked to sign a release form for the release of their personal information. The investigator, or their designee, will review which specific study information will be collected and to whom that information will be disclosed. After speaking with the participant, questions about the details of the release form will be answered. A. Study Protocol Procedures before the first exercise session. Before the first exercise session, a subject's VO2 max will be assessed by following the Bruce protocol, and the basal insulin dose will be determined according to the investigator's usual routine. Exercise sessions. Each participant will be randomly assigned to a study sequence (control, strategy 1, strategy 2, and strategy 3). Each exercise session will be separated by at least 3 days, and participants will be expected to complete all sessions within 12 weeks of the baseline or screening visit. Participants will be advised to avoid any vigorous exercise within 24 hours before or after laboratory-based exercise tests. Subjects will be questioned about their activity to monitor their adherence to these recommendations. The day before each exercise session, participants will: insert a new insulin infusion device in the abdomen or upper gluteal area (make sure it is not in the limb being exercised): for CGM users, insert a new CGM sensor; for non-CGM users, come to the clinic to have the CGM sensor inserted for use with a receiver in a concealed manner. Research staff will contact the participant the night before and the morning of each exercise session to check glucose levels and help mitigate hyperglycemia or hypoglycemia before exercise. If, at the time of the morning call, the glucose concentration is below target (< 100 mg / dL), then supplemental rapid-acting carbohydrates will be taken. If it is above target (> 140 mg / dL), then a conservative correction dose will be recommended (based on the insulin sensitivity factor). The participant will arrive at the facility in a fasted state (minimum 8-hour fast). Blood glucose levels will be checked upon arrival. Oral or IV glucose may be administered to the participant within the interval before exercise. Women will undergo a urine pregnancy test before each exercise session (if applicable). Before an exercise session can begin: (1) The participant's glucose level must be 100 mg to 140 mg per dL (venous, plasma); (2) The last insulin bolus (food time or correction) must be > 3 hours prior. During the exercise sessions. On each visit, participants will perform one of the following four exercise sessions (the sequence of sessions will be determined by random selection): Control study: No adjustment of basal insulin, no carbohydrate intake until glucose falls to < 70 mg / dL. Then 20 grams of dextrose will be given orally. Strategy 1: Reduce basal insulin by up to 50 percent five minutes before the start of exercise. The basal insulin rate will be returned to the usual rate 45 minutes after the start of exercise. Strategy 2: Dextrose tablets are taken orally (20 grams) five minutes before the start of exercise and 30 minutes into exercise (total of 40 grams). Strategy 3: Glucagon (150 pg) five minutes before the start of exercise (upper gluteal (SQ) or abdomen). The dose for Strategy 3 will most likely be 150 pg, but it can be modified to be lower or higher, with a possible range of 75 pg to 300 pg. One or more pilot phases may be conducted before the start of the study to determine the most appropriate dose. The participant's pump will be hidden during the control study, strategy 1, and strategy 3, and a saline injection will be administered during the control study and strategy 1 in such a way as to conceal the strategy from the participant. The exercise will consist of moderate-intensity aerobic activity (jogging or brisk walking on a treadmill) performed at 50 to 55 percent of the previously determined VO2 max for 45 minutes. Continuous heart rate measurements will be taken along with intermittent assessments of perceived exertion (Borg scale: 6-20). The exercise will be terminated if: (i) Glucose is < 70 mg / dL; participants will be treated for hypoglycemia (initially with 20 grams of oral dextrose tablets or IV dextrose or IM glucagon, depending on the severity of the hypoglycemia); or (ii) Participants experience dizziness, pallor, and other symptoms of poor perfusion and / or exercise intolerance. Any treatment given will be recorded. Participants who are unable to complete an exercise session due to termination will still complete the remaining study procedures. Following each exercise session, the participant will rest for 30 minutes, then consume a standardized meal containing 44 to 50 grams of carbohydrates, which constitute approximately 55 percent of the calories, along with approximately 20 percent of the calories from protein and approximately 25 percent of the calories from fat. The bolus insulin dose will be based on the carbohydrate content of the meal and the individualized insulin-to-carbohydrate ratio of the participant. The insulin will be administered five minutes before the standardized meal. In the first phase of the experiments, insulin corrections will not be given unless subjects have hyperglycemia > 270 mg / dL after exercise or before the meal.If hypoglycemia occurs before food, subjects will be treated with 20 grams of fast-acting carbohydrates before food consumption. The participant will be monitored for at least two hours after feeding before release. Participants will be given a food log and another standardized food item to take with them upon release. Participants will be instructed to eat the standardized food item for their next meal and record the time they ate it. They will also be asked to record any food they consume until noon the following day. For participants using the concealed CGM, the sensor will be removed at 12:00 PM the following day at their home (approximately 24 hours after the end of the exercise session). The participant will bring the device to the next visit or return it by mail to the clinic. Participants will receive a phone call the day after each exercise session that results in any adverse event. For CGM users, CGM data will be downloaded at each visit, and arrangements will be made to transmit the data immediately after the last exercise session. The participant's insulin pump data will also be downloaded at each visit, and arrangements will be made to transmit the data immediately after the last exercise session. Sample collection. Blood samples were collected via a venous catheter, where plasma glucose was measured using a YSL analyzer. Blood samples were collected at: baseline (at -30, -15, -5 and 0 minutes), during exercise (at 5, 10, 15, 25, 35 and 45 minutes), during recovery after exercise (at 50, 55, 60 and 75 minutes), and regularly following a standardized mixed meal for 90 minutes (at 90, 105, 120, 135 and 165 minutes).Plasma will be used from the blood samples that were collected to measure the following hormones, pro-inflammatory markers and metabolites at baseline, during exercise, in recovery after exercise, and regularly following the standardized mixed meal: insulin, glucagon, cortisol, growth hormone, catecholamines (epinephrine and norepinephrine), interleukin-6, tumor necrosis factor-α, non-esterified fatty acids, β-hydroxybutyrate and / or lactate. B. Results This study aimed to determine whether administering a mini-dose of glucagon (MDG) just before exercise results in greater glucose stability than (1) basal reduction, (2) pretreatment with glucose tablets, and (3) a control without either of these, for moderate-intensity exercise in patients with T1D. Subjects aged 18 to <65 years with a T1D duration >2 years, with a random C-peptide <0.6 ng / mL, who had used a CSII (insulin pump) for at least 6 months, with no plans to discontinue pump use during the study, and who exercised regularly (>30 minutes of moderate or more vigorous aerobic activity >3 times per week) were randomly selected for a 4-period crossover study. The control group had no basal insulin adjustment and no carbohydrate intake unless glucose was <70 mg / dL (in which case 20 grams of dextrose were given). Reduction of basal rate: The basal rate decreased by 50 percent 5 minutes before the start of exercise (the basal rate returned to the usual rate at the end of exercise). Glucose tablets: Take dextrose tablets orally (20 grams) 5 minutes before the start of exercise, and 20 grams again 30 minutes after the start. Mini dose of giucagon: Giucagon (150 pg) 5 minutes before the start of exercise (SQ-abdomen). The exercise consisted of moderate-intensity aerobic activity (brisk uphill walking on a treadmill) performed at 50–55 percent of VO2 max for 45 minutes. It was performed in a fasted state. The same workload was used in each exercise session. Exercise was terminated if: glucose was <70 mg / dL (the participant was treated for hypoglycemia) and / or the participant experienced dizziness, pallor, and other symptoms of poor perfusion and / or exercise intolerance. Following each exercise session, participants rested for 30 minutes and then consumed a standardized meal. The meal contained 44 to 50 grams of carbohydrates (55 percent of calories from carbohydrates, 20 percent from protein, and 25 percent from fat). Insulin was administered as a pump bolus 5 minutes before the standardized meal using the participants' own carbohydrate ratio, with correction only if glucose was >270 mg / dL. Figures 2 through 6 show the results obtained in such a study. The results show that a medium-density glucose (MDG) bolus given just before aerobic exercise reduces the frequency of a drop in blood glucose concentration in people with type 1 diabetes. Reducing basal insulin at the start of exercise was not effective in limiting the drop in blood glucose during aerobic exercise.Increased carbohydrate intake (40 grams / hour) during exercise increased hyperglycemia during activity and exaggerated post-exercise glucose excursions. Modified carbohydrate intake (MCI) was more effective than insulin reduction in reducing exercise-induced hypoglycemia and resulted in lower post-intervention hyperglycemia than carbohydrate intake alone. MCI provides a novel and effective strategy for reducing exercise-induced hypoglycemia and post-exercise hyperglycemia. ocaonn / nznz / E / Y Table 1 Measurement of CGM in the late recovery period* Control Reduction based! MDG Tablets n = 14 n ~ 13 n -13 n ™ 14 Nadir Glucose, mg / dL · median 45 44 49 51 Average Glucose, mg / dL - median 129 138 134 150 % of time < 70 mg / dL - average 11 % 8% 8% 7% % of time 70-180 mg / dL - average 73 % 71 % 67% 66% Control Basal Reduction MDG Tablets n - 14 n -13 n ~ 13 n - 14 Coefficient of variation - median 32 % 34% 39 % 34 % * Eat standard food until midday the day after the exercise session; no statistically significant differences were found for the above measurements. ocaonn / nznz / E / Y Table 2 Overnight CGM measurements* Control Reduction Baseline! MDG Tablets n = 13 n = 13 n = 11 n = 13 Nadir Glucose, mg / dL - median 102 93 77 91 Average Glucose, mg / dL - median 132 137 128 156 Hypoglycemia Presentation (< 70 mg / dL) - % 23% 23% 27% 23% % of time < 70 mg / dL - average 5% 4% 3% 4% % of time 70-180 mg / dL - average 77% 66% 63% 59% Coefficient of Variation - median 15% 13% 15% 21% * From midnight until 6 am the day after the exercise session; no statistically significant differences were found for the above measurements.

Claims

REIVSNDiCACjQNES 1. A method for the treatment of exercise-associated hypoglycemia in a subject, comprising the administration of a glucagon composition or a glucagon analogue 5 to 30 minutes before the start of exercise.

2. The method according to what is claimed in claim 1, wherein the subject is a diabetic subject.

3. The method according to what is claimed in claim 1, wherein 50 pg to 300 ug of glucagon or a glucagon analogue, preferably 150 g, are administered.

4. The method according to claim 1, wherein the glucagon or glucagon analogue is administered as a bolus.

5. The method according to claim 1, wherein the glucagon or glucagon analogue is administered as an infusion over 5 to 45 minutes.

6. The method according to any of claims 1 to 5, wherein the glucagon or glucagon analogue is administered from a glucagon or glucagon analogue delivery apparatus.

7. The method according to claim 6, wherein the glucagon delivery apparatus comprises: (i) a reservoir containing the glucagon composition or a glucagon analogue; and (ii) an electronic pump configured to deliver, via the intradermal, subcutaneous, or intramuscular route, at least a portion of the composition to a subject.

8. The method according to what is claimed in any of claims 1 to 7, which further comprises monitoring the subject's blood glucose levels.

9. The method according to what is claimed in any of claims 6 to 8, wherein the apparatus is a closed-loop system for delivering glucagon to the patient.

10. The method according to what is claimed in any of claims 6 to 8, wherein the apparatus is an open-loop system for delivering glucagon to the patient.

11. The method according to what is claimed in any of claims 6 to 8, wherein the apparatus is a non-loop system for supplying glucose to the patient.

12. The method according to what is claimed in any of claims 1 to 11, wherein the glucagon or glucagon analogue composition is a single-phase solution comprising glucagon, the glucagon analogue, or a salt form of any of the same, dissolved in a non-aqueous solvent.

13. The method according to claim 12, wherein the non-aqueous solvent ocaonn / nznz / E / Y is a polar aprotic solvent.

14. The method according to claim 13, wherein the aprotic solvent is DMSO. 1o. The method according to claim 13, wherein the aprotic solvent is a deoxygenated aprotic solvent.

16. The method according to claim 13, wherein the glucagon or glucagon analogue composition further comprises an ionization-stabilizing excipient, wherein: (i) the glucagon, glucagon analogue, or a salt thereof, is dissolved in the aprotic solvent in an amount of approximately 0.1 mg per mL, up to the solubility limit of glucagon, glucagon analogue, or a salt thereof, and (ii) the ionization-stabilizing excipient is dissolved in the aprotic solvent in an amount sufficient to stabilize the ionization of glucagon, glucagon analogue, or a salt thereof.

17. The method according to what is claimed in claim 16, wherein the ionization stabilizing excipient is at a concentration of 0.1 mM to less than 100 mM.

18. The method according to what is claimed in claim 16, wherein the ionization stabilizing excipient is a mineral acid.

19. The method according to what is claimed in claim 18, wherein the mineral acid is sulfuric acid.

20. The method according to what is claimed in claim 16, wherein the ionization stabilizing excipient is sulfuric acid and the aprotic solvent is DMSO.

21. The method according to what is claimed in claim 16, wherein the composition of glucagon or a glucagon analogue has a moisture content of less than 10, 5 or 3 percent.

22. The method according to what is claimed in claim 16, wherein the composition of giucagon or an analogue of giucagon further comprises a preservative in less than 10, 5 or 3 percent by weight / volume.

23. The method according to what is claimed in claim 22, wherein the preservative is benzyl alcohol.

24. The method according to what is claimed in claim 16, wherein the composition further comprises a sugar alcohol in less than 10, 5 or 3 percent by weight / volume.

25. The method according to what is claimed in claim 24, wherein the sugar alcohol is mannitol.

26. The method according to what is claimed in claim 16, wherein the composition of glucagon or a glucagon analogue further comprises one or more carbohydrates.

27. The method according to what is claimed in claim 26, wherein the carbohydrate is trehalose and / or mannitol.

28. The method according to what is described in any of claims 16 to 27, wherein the composition of glucagon or a glucagon analogue comprises at least 80 percent by weight of the polar aprotic solvent, 3 to 7 percent by weight of the carbohydrate, 0.001 to 0.1 percent by weight of an amphoteric molecule, and 0 percent by weight to less than 0.1 percent by weight of the acid.

29. The method according to what is claimed in any of claims 1 to 28, wherein the composition of glucagon or a glucagon analogue has a water content of 0 to less than 15 percent by weight, 0 to less than 3 percent by weight, 3 to 10 percent by weight, or 5 to 8 percent by weight.

30. The method according to any one of claims 1 to 29, wherein the glucagon, glucagon analogue, or salt form thereof, has been previously dried from a buffer, wherein the dried glucagon, glucagon analogue, or salt form thereof has a first ionization profile corresponding to optimum stability and solubility for the glucagon, glucagon analogue, or salt form thereof, wherein the dried glucagon, glucagon analogue, or salt form thereof is reconstituted in a polar aprotic solvent and has a second ionization profile in the polar aprotic solvent, and wherein the first and second ionization profiles are within 1 pH unit of each other.

31. The method according to what is claimed in any of claims 1 to 30, wherein the glucagon composition or a glucagon analogue has been stored in the reservoir for at least 1, 2, 3, 4, 5, 6, 7, 14, 21, 30, 45 or 60 days.