Sustained release injectable formulations using aprotic polar solvents

Abstract

The present invention relates to the use of aprotic polar solvents, water and ionization stabilizers for the preparation of stable therapeutic formulations, in particular sustained release formulations, of various active pharmaceutical ingredients (APIs), such as levothyroxine. In particular, the present invention relates to the use of an aprotic polar solvent and at least one ionizing stabilizer in the preparation of a stable therapeutic formulation of a plurality of APIs by dissolving at least one API in an aprotic polar solvent system to produce a stable sustained release therapeutic formulation suitable for the treatment, prevention and / or diagnosis of diseases or physical conditions in humans and veterinary animals. The invention also provides methods of making such formulations.

Description

[0001] Cross - reference to related applications and incorporation by reference

[0002] This application claims the benefit of U.S. Provisional Application No. 63 / 380,213, filed Oct. 19, 2022, which is incorporated herein by reference in its entirety. Background of the Invention A. Technical Field

[0004] The present invention pertains to the fields of medicine and pharmacy. Certain embodiments generally relate to therapeutic aprotic solvent formulations comprising one or more active pharmaceutical ingredients that can be used as therapeutic formulations, particularly sustained - release formulations, for the treatment, prevention, and / or diagnosis of diseases, disorders, and medical conditions in mammals, particularly humans. In particular, the present invention relates to the use of aprotic polar solvents and at least one ionization stabilizer in the preparation of stable therapeutic formulations of multiple active pharmaceutical ingredients (APIs) by dissolving at least one API in an aprotic polar solvent system to produce stable sustained - release therapeutic formulations that can be used for the treatment, prevention, and / or diagnosis of diseases or physical conditions in humans and veterinary animals. B. Background Art

[0005] Parenteral formulations of therapeutic molecules (such as active pharmaceutical ingredients, e.g., therapeutic peptides, therapeutic proteins, and therapeutic small molecules) prepared in aprotic polar solvent systems (e.g., DMSO - based solvent systems) have improved drug molecule stability due to the absence of water - mediated degradation pathways. These pathways, including hydrolysis, deamidation, and aspartic acid isomerization, are known to largely contribute to the instability of peptides and proteins in water - based formulations. Additionally, hydrolysis is known to promote the chemical instability of small molecule drugs. Existing studies have demonstrated that low - moisture peptide and small molecule formulations have improved stability in DMSO relative to aqueous solutions (see, e.g., U.S. Pat. Nos. 9,339,545, 10,485,850, and 11,020,403, the entire disclosures of which are incorporated herein by reference).

[0006] However, one disadvantage of using such formulations is that, without the addition of certain release modifiers, they do not always promote the sustained-release pharmacokinetics of the subject to whom such formulations are administered for treating, preventing, or diagnosing a disease or medical condition. Sustained-release formulations are known in the art, and there is drug precipitation at the injection site. Generally, these formulations include release modifiers that induce drug precipitation or co-precipitation and release modifiers that create a depot at the injection site. Examples are formulations of certain polymers (such as PLGA) and formulations of drugs dissolved in DMSO (see, for example, US 2021 / 0401945A1, the disclosure of which is incorporated herein by reference in its entirety). After injection, the water-insoluble PLGA encapsulates the soluble drug. The therapeutic compound diffuses from the PLGA matrix into the surrounding tissue and the patient's bloodstream, and then the result is the sustained-release pharmacokinetics of the administered drug. Thus, to date, the development of sustained-release formulations of therapeutic compounds in non-aqueous liquid systems has mainly focused on adding certain such release modifiers to the drug formulation to produce a sustained-release effect. These release modifiers can be polymer molecules, microparticles, liposomes, etc. However, the use of release modifiers can be expensive, thereby increasing the cost of the therapeutic formulation to manufacturers, distributors, healthcare providers, and patients and to the healthcare system in general. The use of release modifiers may also result in the need to administer large injection volumes, and large injection volumes tend to cause discomfort or pain to the patient, introduce stability problems, have toxic or allergic effects on patients receiving the formulation containing the release modifier, etc.

[0007] Accordingly, there is a need in the art for stable, ready-to-use sustained-release therapeutic formulations that overcome the problems associated with the previously available sustained-release formulations of therapeutic agents. Such formulations are not only storage-stable but also can be provided for self-administration by patients in an outpatient setting for once-weekly (or even less frequent) treatment formulations for treating a variety of medical conditions and diseases that are typically treated with daily (or more frequent) administrations. Such formulations have several advantages compared to currently commercially available formulations, which include injecting the formulation by a route that bypasses the digestive tract (such as subcutaneously), thereby avoiding the absorption difficulties of oral formulations of certain therapeutic agents currently in use, including oral sustained-release formulations, and potentially improving the patient experience and patient compliance compared to daily oral therapy or injecting a sustained-release formulation containing a release modifier. In addition, such formulations can provide enhanced solubility of therapeutic agents that are typically only sparingly soluble or insoluble in a typical aqueous solvent system, as well as more long-term storage stability of the resulting formulation, thereby improving the economic efficiency of these therapeutic formulations compared to other commercial products. SUMMARY OF THE INVENTION

[0008] The embodiments described herein provide formulations that contain multiple pharmaceutical ingredients and meet the needs mentioned above in the art. In other embodiments, the present invention also provides methods for producing such formulations, particularly sustained-release formulations, and methods of use of such formulations for treating, preventing, and / or diagnosing various diseases, disorders, and medical conditions in humans and veterinary animals. A significant advantage of certain embodiments of the present invention is that it is not necessary to include a release modifier in an injectable formulation to create a sustained-release formulation of a therapeutic agent. Thus, the formulation is inherently easy to produce, thereby providing an economic advantage to manufacturers and users of such formulations. In addition, when such a formulation is administered to a patient, a relatively low injection volume can be maintained, improving the patient experience and compliance. In principle, the present invention relies on the inherent stability of molecules that are highly soluble in DMSO (and other polar aprotic solvent systems) but have relatively poor solubility in an aqueous environment, such as the interstitial fluid in the body where the formulation is injected, such as a subcutaneous or intramuscular space.

[0009] The embodiments described herein provide storage-stable compositions (formulations) that contain one or more therapeutically active ingredients (e.g., one or more active pharmaceutical ingredients) that are highly soluble (i.e., at least about 50% soluble, preferably at least about 75% soluble, at least about 80% soluble, at least about 85% soluble, at least about 90% soluble, at least about 95% soluble, or at least about 99% soluble) in a polar aprotic solvent system as described herein (and particularly in a DMSO-containing solvent system), and are poorly soluble (i.e., less than about 50% soluble, less than about 40% soluble, less than about 30% soluble, less than about 25% soluble, less than about 20% soluble, less than about 15% soluble, less than about 10% soluble, less than about 5% soluble, or less than about 1% soluble) in an aqueous environment, such as the interstitial fluid in a patient in whom the formulation has been injected, such as a subcutaneous or intramuscular space. In certain embodiments, the formulation contains a therapeutic peptide or a therapeutic small molecule, such as levothyroxine or a salt thereof, benzodiazepines (e.g., diazepam), tamsulosin, and the like. In a preferred embodiment, the active pharmaceutical ingredient is present in the formulation at a sufficient concentration such that the active pharmaceutical ingredient is released continuously into the bloodstream of a patient who has been injected with the formulation. In other embodiments, the present invention provides methods for preparing such storage-stable sustained-release therapeutic formulations. In other embodiments, the present invention provides methods of using the storage-stable sustained-release formulations of the present invention in methods for treating, preventing, and / or diagnosing diseases, medical conditions, or disorders in animals, including veterinary animals and humans, that are suffering from or are susceptible to certain diseases, medical conditions, or disorders. In other embodiments, the present invention provides methods of using the storage-stable sustained-release formulations of the present invention in methods for treating, preventing, and / or diagnosing diseases, medical conditions, or disorders in animals, including veterinary animals and humans, that are suffering from or are susceptible to certain diseases, medical conditions, or disorders.

[0010] In certain exemplary embodiments, the present invention provides storage-stable sustained-release levothyroxine formulations, which can be used for treating and / or preventing hypothyroidism, particularly severe hypothyroidism such as myxedema or myxedema coma / crisis, and can be used as an adjunct to certain other therapeutic procedures. In other exemplary embodiments, the present invention provides storage-stable sustained-release benzodiazepine drug formulations, such as storage-stable sustained-release formulations of diazepam, which can be effectively used for treating and / or preventing diseases or conditions such as anxiety disorders, muscle spasms, and seizures, and can be effectively used as an adjunct to certain other therapeutic procedures. In other exemplary embodiments, the present invention provides storage-stable sustained-release tamsulosin formulations, which can be used for treating and / or preventing benign prostatic hyperplasia and kidney stones, and can be used as an adjunct to certain other therapeutic procedures.

[0011] Such formulations suitably utilize the same non-aqueous formulation technology (XeriSol TM ; Xeris Pharmaceuticals, Inc.; Chicago, IL) as the currently commercially available rapid-release glucagon emergency product ( Xeris Pharmaceuticals) to address the stability and solubility challenges of certain aqueous compositions containing small molecules or aqueous compositions containing peptides. Such storage-stable sustained-release formulations of the present invention are clear, ready-to-use, non-aqueous solutions before injection. After administration, for example, after subcutaneous administration, the small molecule or therapeutic peptide forms a depot due to its low solubility under physiological conditions, promoting the gradual release of the therapeutic molecule into the bloodstream and improving the bioavailability, pharmacokinetics, and in some cases, the sustained release of the therapeutic molecule compared to orally administered compositions or rapid-release parenteral compositions.

[0012] Accordingly, in certain embodiments, the present invention provides a therapeutic formulation comprising: (a) at least one therapeutic agent, (b) at least one ionization-stabilizing excipient, and (c) an aprotic polar solvent. In particular, the formulation can be stably stored for at least two years under refrigerated conditions (2°C to 10°C), and when administered to a patient, the formulation results in an extended period of time (i.e., sustained release of the therapeutic agent) of the therapeutic level of the therapeutic agent being present in the patient's blood as compared to an immediate-release formulation comprising the same therapeutic agent. According to the present invention, any therapeutic agent that is poorly soluble in an aqueous environment (such as interstitial fluid in a human or veterinary animal) but is soluble or highly soluble in an aprotic polar solvent system (such as a solvent system comprising DMSO) can be advantageously used in the formulations of the present invention. In certain such embodiments, the therapeutic agent is a small molecule or a salt thereof, which can be any small molecule therapeutic agent. Examples of such small molecule drugs include, but are not limited to, levothyroxine, sumatriptan, ketorolac, and ondansetron. In other such embodiments, the therapeutic agent is a peptide such as glucagon-like peptide, a glucagon analog, a glucagon mimetic, or a salt thereof.

[0013] Dissolve at least one ionization-stabilizing excipient in the aprotic solvent in an amount that stabilizes the ionization of the therapeutic agent. In certain aspects, the concentration of the ionization-stabilizing excipient is from 0.01 mM to less than 200 mM. The ionization-stabilizing excipient can be a proton donor (such as an acid) and / or a proton acceptor (such as a base) component. The ionization-stabilizing excipient can be, but is not limited to, a mineral acid. In certain embodiments, the mineral acid can be selected from hydrochloric acid, sulfuric acid, nitric acid, and phosphoric acid. The ionization-stabilizing excipient can also be an organic acid (acids having a carboxylic -COOH functional group). Non-limiting examples of organic acids include acetic acid, citric acid, malic acid, lactic acid, and amino acids. In certain aspects, the aprotic solvent is DMSO. In a particular aspect, the ionization-stabilizing excipient is a mineral acid, particularly sulfuric acid or hydrochloric acid, and the aprotic solvent is DMSO.

[0014] In other embodiments, the present invention provides a method of treating, ameliorating, or preventing a disease, physical condition, or disorder in a patient, such as a veterinary animal or a human, who has or is susceptible to the disease, physical condition, or disorder. Suitable such methods according to certain aspects of the present invention include introducing an effective amount of the storage-stable sustained-release formulation of the present invention into a patient in need thereof in a manner suitable for promoting the release of the therapeutic compound from the formulation into the patient's bloodstream. In certain such embodiments, the formulation is introduced into the subject by parenteral administration, such as by injection (which can be subcutaneous, intradermal, or intramuscular) or infusion (which can be intravenous infusion, or can be accomplished by pump infusion, such as by continuous pump infusion or bolus pump infusion or a combination thereof). In certain aspects, the present invention provides a method of treating or preventing human hypothyroidism by administering to a human the storage-stable sustained-release levothyroxine formulation of the present invention.

[0015] In other embodiments, the present invention provides a method for preparing a storage-stable sustained-release therapeutic formulation, the method comprising mixing at least one ionization-stabilizing excipient, an aprotic polar solvent, and at least one therapeutic agent, particularly at least one therapeutic agent that is poorly soluble in an aqueous environment and soluble or highly soluble in an aprotic polar solvent system, thereby forming a storage-stable sustained-release therapeutic formulation which, when administered to a patient, results in the therapeutic agent at a therapeutic level being present in the blood of the patient for an extended period of time in some cases as compared to an immediate-release formulation containing the same therapeutic agent.

[0016] To produce the storage-stable sustained-release therapeutic formulation of the present invention, at least one ionization-stabilizing excipient can be dissolved in the aprotic solvent in an amount sufficient to stabilize the ionization of the therapeutic agent. Suitable such ionization-stabilizing excipients (including but not limited to mineral acids) and the desired concentrations included in the formulations of the present invention include those described in detail elsewhere herein.

[0017] The formulations of the present disclosure can also contain less than 10% weight / volume, 5% weight / volume, or 3% weight / volume of a preservative. In certain embodiments, the preservative is benzyl alcohol.

[0018] The formulations of the present disclosure can also contain one or more disaccharides in an amount less than 10% weight / volume, 5% weight / volume, or 3% weight / volume. In certain embodiments, the disaccharide is about 5.5% weight / volume of trehalose dihydrate.

[0019] The formulations can also contain one or more sugar alcohols in an amount less than 10% weight / volume, 5% weight / volume, or 3% weight / volume. In certain embodiments, the sugar alcohol is about 2.9% weight / volume of mannitol.

[0020] In certain embodiments, the freezing point of the formulation can be about 10 °C or less than 10 °C, such as about 10 °C, about 5 °C, about 0 °C, or less than about 0 °C, such as less than -20 °C or -50 °C to -70 °C.

[0021] Therapeutic molecules generally require an optimal or beneficial ionization profile in order to exhibit extended stability when dissolved in an aprotic polar solvent system. Maintaining a beneficial ionization profile of a therapeutic molecule dissolved in an aprotic polar solvent system can be achieved by using at least one ionization-stabilizing excipient. In some aspects, it is not necessary to pre-dry the therapeutic molecule from a buffered aqueous solution prior to reconstitution in the aprotic polar solvent system. The ability to use existing (e.g., commercially available) equipment and the ability to circumvent the need to dry therapeutic molecules (e.g., peptides) from buffered aqueous solutions can save considerable time and cost at multiple product development stages.

[0022] A stable solution of one or more therapeutic agents dissolved in a non-aqueous aprotic polar solvent (e.g., DMSO) can be prepared by adding a certain amount of a compound or combination of compounds that act as ionization-stabilizing excipients. Without wishing to be bound by theory, it is believed that the ionization-stabilizing excipient can act as a proton source in the aprotic polar solvent system (e.g., a molecule that can donate a proton to the therapeutic molecule), protonating the ionizable groups on the therapeutic molecule, such that the therapeutic molecule has an ionization profile with improved physical and chemical stability in the aprotic polar solvent system. Alternatively, the ionization-stabilizing excipient acts as a proton sink (e.g., a molecule or moiety that can accept / remove a proton from the therapeutic molecule), such that the therapeutic molecule has an ionization profile with improved physical and chemical stability in the aprotic polar solvent system.

[0023] Certain embodiments relate to formulations comprising an ionization-stabilizing excipient at a concentration of at least, at most, or about 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, 1000 mM, or up to the solubility limit of the ionization-stabilizing excipient in the aprotic polar solvent system. In certain aspects, the ionization-stabilizing excipient concentration is from about 0.1 mM to about 100 mM, particularly from about 1 mM to about 10 mM, such as about 1 mM, about 2 mM, about 3 mM, about 4 mM, about 5 mM, about 6 mM, about 7 mM, about 8 mM, about 9 mM, and about 10 mM. In certain embodiments, the ionization-stabilizing excipient can be a suitable mineral acid, such as hydrochloric acid, sulfuric acid, nitric acid, etc. In certain aspects, the ionization-stabilizing excipient can be an organic acid, such as an amino acid, an amino acid derivative, or a salt of an amino acid or amino acid derivative (examples include glycine, trimethylglycine (betaine), glycine hydrochloride, and trimethylglycine (betaine) hydrochloride). In other aspects, the amino acid can be glycine or the amino acid derivative trimethylglycine. In certain aspects, the peptide is less than 150, 100, 75, 50, or 25 amino acids. In other aspects, the aprotic solvent system includes DMSO. The aprotic solvent can be deoxygenated, such as deoxygenated DMSO. In certain embodiments, the formulation can be prepared by the steps of: first adding the ionization-stabilizing excipient to the aprotic polar solvent system, and subsequently adding the therapeutic molecule. Alternatively, the therapeutic molecule can first be dissolved in the aprotic polar solvent system, and then the ionization-stabilizing excipient is added. In other aspects, the ionization-stabilizing excipient and the therapeutic molecule can be dissolved simultaneously in the aprotic polar solvent system. In certain aspects, the therapeutic agent is glucagon, a glucagon analog, or a salt thereof.

[0024] Other embodiments of the present invention relate to methods of stably formulating therapeutic agents (e.g., peptides or small molecules), which include: (a) calculating or determining a suitable ionization-stabilizing excipient (e.g., proton concentration) required to achieve a stable ionization profile of a target therapeutic agent (e.g., one or more peptides or one or more small molecules) in an aprotic polar solvent system; (b) mixing at least one ionization-stabilizing excipient with the aprotic polar solvent system to obtain a suitable ionization environment that provides the ionization profile determined in (a); and (c) dissolving one or more target therapeutic agents in an aprotic solvent having a suitable environment for physically and chemically stabilizing the therapeutic agent. In certain non-limiting aspects, at room temperature, at refrigerated temperatures (e.g., from about 2°C to about 10°C, or from about 2°C to about 8°C, or about 2°C, about 3°C, about 4°C, about 5°C, about 6°C, about 7°C, about 8°C, about 9°C or about 10°C), or at sub-zero temperatures (e.g., from about -4°C to about -80°C, or about -4°C, about -10°C, about -15°C, about -20°C, about -25°C, about -40°C, about -45°C, about -50°C, about -60°C, about -70°C or about -80°C), the therapeutic agent is chemically or physically stable for at least or about 0.25 years, about 0.5 years, about 1 year, about 2 years, about 3 years, about 4 years or about 5 years, and more preferably from about 0.25 years to about 3 years, and even more preferably from at least about 6 months to about 2 years. In certain aspects, the dissolution of the therapeutic agent and the addition of the ionization-stabilizing excipient to the aprotic polar solvent system can be carried out in any order or simultaneously. However, the inventors have found that in order to maximize the dissolution, stability, and / or bioactivity of some therapeutic agents such as levothyroxine, it is preferred to first mix the ionization-stabilizing excipient with the aprotic polar solvent and then dissolve the therapeutic agent in the mixture. However, other formulations of the present invention can be prepared by first dissolving the therapeutic agent in an aprotic polar solvent and then adding the ionization-stabilizing excipient to the solution shortly thereafter (e.g., within about 5 minutes), or the ionization-stabilizing excipient and the therapeutic agent can be added to or dissolved in the aprotic polar solvent system simultaneously. One or more other formulation components (e.g., sustained-release modifiers (e.g., polymeric compounds such as poly(lactic-co-glycolic acid) or PLGA; see U.S. Patent Publication No. US2021 / 0401945A1, which is incorporated herein by reference in its entirety), preservatives, surfactants, polysaccharides, sugar alcohols, etc.) can be incorporated into the formulation before or after the addition of the therapeutic agent. The concentration of the therapeutic agent and / or ionization-stabilizing excipient added to the solution can be between 0.01 mM, 0.1 mM, 1 mM, 10 mM, 100 mM, 1000 mM, or up to its solubility limit and includes all values and all ranges described in more detail elsewhere herein. In certain aspects, the aprotic polar solvent system is deoxygenated. In other aspects, the aprotic polar solvent in the solvent system comprises, consists essentially of, or consists of DMSO or deoxygenated DMSO.

[0025] In other aspects of the present invention, methods for treating or preventing a condition, disease, disorder, etc. are disclosed, which include administering an effective amount of one or more formulations of the present invention to a subject in need thereof to treat or prevent a condition, disease, disorder, etc. Any suitable dose of a therapeutic agent (e.g., a protein, peptide, or small molecule) can be administered in the methods of the present invention. Of course, the dose administered will vary depending on known factors such as the pharmacodynamic properties of the particular composition, salt, or combination thereof; the age, health, or weight of the subject; the nature and degree of the symptoms; the metabolic properties of the drug and the patient, the type of concurrent treatment; the frequency of treatment; or the desired effect. In certain aspects, hypothyroidism can be treated or prevented by administering a formulation described herein that contains an effective amount of levothyroxine. In other aspects, hypoglycemia can be treated by administering a formulation described herein that contains an effective amount of glucagon.

[0026] The stable formulations described herein can be used for parenteral injection of any therapeutic agent (protein, peptide, and / or small molecule) that has limited or poor stability or solubility in an aqueous environment. In certain aspects, the formulations described herein are provided in the form of injectable formulations. The injectable formulations can be administered into the epidermal layer, dermal layer, subcutaneous layer, or intramuscular layer of a patient. In certain aspects, the formulation is administered into the skin, particularly by intradermal or subcutaneous administration.

[0027] Definitions

[0028] The terms "container", "reservoir", "infuser", "pump", "formulation flow path", "fluid flow path", etc. should be interpreted as interchangeable and equivalent, as these components will come into direct contact with the formulation being administered or stored and have the potential to interact with the components and their surfaces. The term means any and all components that the formulation will contact during storage (e.g., pump reservoir) and delivery (e.g., fluid flow path in the pump and infuser when in series with the pump). As used herein, the term "infuser" can be interpreted to include an internal infuser (i.e., the infuser contained within a patch pump) as well as the complete tubing system that connects the pump to the pump user and is typically located outside the pump. In certain configurations, the external infuser includes a cannula (e.g., for subcutaneous administration), an adhesive mount, a quick disconnect, and a pump barrel connector (e.g., a Luer type connector).

[0029] The terms "formulation" and "composition" are used interchangeably herein and, as used herein, refer to a mixture of at least two components to produce a formulation that contains all of these components, some of these components, or a complex or reaction mixture or a reaction resulting from the mixture of the components.

[0030] As used herein, the term "dissolution" refers to the process by which one or more materials in a gaseous, solid, or liquid state become one or more solutes, one or more dissolved components of a solvent, forming a solution of a gas, liquid, or solid in the solvent. In certain aspects, a therapeutic agent or excipient, such as an ionization-stabilizing excipient or a sustained-release modifier or other component, is present in an amount up to its solubility limit or is completely dissolved. The term "dissolution" refers to the incorporation of a gas, liquid, or solid into a solvent to form a solution.

[0031] As used herein, the term "elastomer" refers to a natural or synthetic polymer having elastic properties. The terms "elastomer" and "rubber" are used interchangeably herein.

[0032] As used herein, the term "excipient" refers to a natural or synthetic substance (a component of an inactive ingredient) formulated with the active or therapeutic ingredient of a drug, which includes stabilizing, swelling, or therapeutically enhancing the active ingredient in the final dosage form, such as facilitating drug absorption, reducing viscosity, enhancing or reducing aqueous or non-aqueous solubility, adjusting tonicity, alleviating injection site discomfort, lowering the freezing point, or enhancing stability. In addition to contributing to in vitro stability, such as preventing denaturation or aggregation during the intended storage period, excipients can also be used in the manufacturing process to facilitate handling of the relevant active substance, such as promoting powder flowability or non-sticky properties.

[0033] In the context of the present invention, a "small molecule drug" is a bioactive compound (and its salts) that can bring about a desired, beneficial, and / or pharmacological effect in a subject. These "small molecule drugs" are organic or inorganic compounds. Thus, in the context of the present invention, small molecule drugs are not polymeric compounds. Generally, small molecule drugs have a molecular weight of less than about 1000 daltons. Some small molecule drugs are "moisture-sensitive" because they become increasingly unstable in the presence of water. Similarly, salts that can be used with small molecule drugs are known to those skilled in the art and include salts formed with inorganic acids, organic acids, inorganic bases, or organic bases.

[0034] The term "therapeutic agent" or "therapeutic drug" encompasses proteins, peptides, small molecule drugs, and their pharmaceutically acceptable salts. Useful salts are known to those skilled in the art and include salts formed with inorganic acids, organic acids, inorganic bases, or organic bases. Therapeutic agents useful in the present invention are those proteins, peptides, and small molecule compounds that, when administered to a human or animal, either alone or in combination with other pharmaceutical excipients or inert ingredients, affect a desired beneficial and generally pharmacological effect. Therapeutic agents may also be interchangeably described herein as "active pharmaceutical ingredients" ("API").

[0035] The terms "peptide" and "peptide compound" refer to amino acid polymers or peptidomimetic (peptide mimic) polymers of up to about 200 amino acid residues joined together by amide bonds (CONH) or other bonds. In some aspects, a peptide can be up to 150, 100, 80, 60, 40, 20, or 10 amino acids. "Protein" and "protein compound" refer to polymers of more than 200 amino acid residues joined together by amide bonds. These terms include analogs, derivatives, agonists, antagonists, and pharmaceutically acceptable salts of any peptide compound or any protein compound disclosed herein. The terms also include peptides, proteins, peptide compounds, and protein compounds having in their structure a modified, derivatized, or naturally occurring D-amino acid and / or a peptidomimetic unit of an amino acid having a D-configuration or an L-configuration in part.

[0036] "Analog" and "mimetic," when referring to a peptide or a protein, refer to a modified peptide or protein in which one or more than one amino acid residue has been replaced by other amino acid residues, or in which one or more than one amino acid residue has been deleted from the peptide or protein, or in which one or more than one amino acid residue has been added to the peptide or protein, or any combination of such modifications. Such addition, deletion, or replacement of amino acid residues can occur at any point or points along the primary structure of the peptide, including at the N-terminus and / or the C-terminus of the peptide or protein.

[0037] "Derivative" refers to a parent peptide or a parent protein or an analog thereof that has been chemically modified relative to the parent peptide or the parent protein, where at least one substituent is not present in the parent peptide or the parent protein or its analog. One such non-limiting example is a parent peptide or a parent protein that has been covalently modified. Typical modifications are amide, sugar, alkyl, acyl, ester, polyethylene glycolylation, etc.

[0038] "Single-phase solution" refers to a solution prepared by dissolving a therapeutic agent in a solvent or a solvent system (e.g., a mixture of two or more than two solvents (such as a solvent and a co-solvent)), where the therapeutic agent is completely dissolved in the solvent or the solvent system and there is no longer any visible particulate matter, such that the solution can be described as optically clear. A single-phase solution can 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 (such as powder) suspended in a fluid.

[0039] The term "inhibit" or "reduce" or any variant of these terms includes any measurable reduction or complete inhibition to achieve a desired result.

[0040] "Effective" or "therapeutic" or "preventive" or any variant of these terms means sufficient to accomplish a desired, intended, or targeted result.

[0041] When referring to a therapeutic agent, "chemical stability" means the formation of an acceptable percentage of degradation products through chemical pathways such as oxidation and / or hydrolysis and / or cleavage and / or other chemical degradation pathways. In particular, if after storage at the product's intended storage temperature (e.g., refrigerated storage or sub-zero storage) for at least one year; or the product is stored at accelerated conditions (25°C / 60% relative humidity) for one month, two months or preferably three months, and no more than about 20% of the degradation products are formed, then the formulations of the type described herein are considered to be chemically stable. In some embodiments, the chemically stable formulations form less than 30%, less than 25%, less than 20%, less than 15%, less than 10%, less than 5%, less than 4%, less than 3%, less than 2% or less than 1% of the degradation products after an extended storage time at the product's intended storage temperature.

[0042] When referring to a therapeutic agent, "physical stability" means the formation of an acceptable percentage of aggregates (e.g., dimers, trimers and larger forms). In particular, if after storage at the product's expected storage temperature (e.g., refrigerated storage or sub-zero storage) for at least one year; or the product is stored at 25°C / 60% relative humidity for one month, two months and preferably at least three months, and no more than about 15% of the aggregates are formed, then the formulation is considered to be physically stable. In some embodiments, the physically stable formulations form less than 15%, less than 10%, less than 5%, less than 4%, less than 3%, less than 2% or less than 1% of the aggregates after an extended storage time at the expected product storage temperature.

[0043] A "stable formulation" means a formulation in which at least about 65% of the therapeutic agent (e.g., small molecule, peptide or its salt) remains chemically and physically stable after storage at room temperature for at least one month, or remains stable after storage at refrigerated temperature or sub-zero temperature for up to one year. Those formulations in which at least about 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% of the therapeutic agent remains chemically and physically stable under these storage conditions are particularly preferred. Those stable formulations that do not exhibit degradation after sterilizing irradiation (e.g., gamma rays, beta rays or electron beams) are especially preferred.

[0044] As used herein, "parenteral injection" refers to the administration of a therapeutic agent (e.g., a peptide or small molecule) by a route other than through the digestive tract - any administration that does not pass through the digestive tract - for example, intravenous infusion, intranasal administration, oral administration, transdermal administration, or injection into or through one or more layers of skin or mucous membrane of an animal such as a human. Standard parenteral injection refers to delivery into the subcutaneous tissue, intramuscular tissue, or intradermal tissue of an animal such as a human. These deeper sites are targeted because the tissue is more easily distended relative to the superficial dermal site to accommodate the injection volume required to deliver most therapeutic agents, for example, from 0.1 cc (mL) to 3.0 cc (mL).

[0045] The term "intradermal" encompasses administration to the epidermal skin layer or the subcutaneous skin layer.

[0046] As used herein, the term "aprotic polar solvent" refers to a polar solvent that does not contain acidic hydrogens and thus does not act as a hydrogen bond donor. Aprotic polar solvents include, but are not limited to, dimethyl sulfoxide (DMSO), dimethylformamide (DMF), ethyl acetate, N-methylpyrrolidone (NMP), dimethylacetamide (DMA), and propylene carbonate.

[0047] As used herein, the term "aprotic polar solvent system" refers to a solution in which the solvent is a single aprotic polar solvent (e.g., pure DMSO) or a mixture of two or more aprotic polar solvents (e.g., a mixture of DMSO and NMP), or a mixture of at least one aprotic polar solvent and at least one other pharmaceutically acceptable solvent system, a non-limiting example being a solvent system comprising an aprotic polar solvent (e.g., DMSO) and a protic polar solvent (e.g., propylene glycol). In other aspects, the term "aprotic polar solvent system" refers to a solution in which the solvent is a mixture of one or more aprotic polar solvents and a quantity of water such as water in a volume / volume ratio of at least about 99.9% aprotic solvent to about 0.1% water, up to a volume / volume ratio of at least about 50% aprotic solvent to about 50% water.

[0048] As used herein, "residual moisture" may refer to the residual moisture (usually residual water) in the drug powder after preparation by the manufacturer / supplier. Typical powders generally have a residual water content of up to 10 weight / weight %. When these powders are dissolved in an aprotic polar solvent system, the residual moisture in the powder incorporates into the formulation. Additionally, the aprotic polar solvent may also contain a certain level of residual moisture. For example, a newly opened bottle of USP-grade DMSO can contain up to 0.1 weight / weight % of water. Residual moisture is different from "added moisture", which is water intentionally added to the formulation, for example, to act as a co-solvent or to lower the freezing point of the aprotic polar solvent system. Moisture can also be introduced into the formulation during the addition of an ionization-stabilizing excipient (e.g., by adding a mineral acid from an aqueous stock solution (e.g., 1N HCl or H 2 SO 4 )) or during the addition of water (e.g., water for injection). The total water content (weight / weight %, unless otherwise stated) in the formulation just after preparation is due to the combined effect of residual moisture and added moisture. Such formulations can still be defined as "non-aqueous", and for the purposes of this disclosure, the formulation encompasses formulations containing less than about 50% water.

[0049] As used herein, "device flow path" refers to the part of the device that can come into contact with the formulation / solution / solvent during the use of the device to administer the formulation / solution / solvent to a subject. In some aspects, the device can be an infusion set in series with a pump that is capable of parenterally administering the formulation / solution / solvent to the subject through various needles and / or tubes. In other aspects, the device can be a patch pump that is directly attached to the patient and does not require the use of an external infusion device in series with the pump.

[0050] The term "about" or "approximately" or "substantially invariant" is defined as being close to as understood by a person of ordinary skill in the art, and in a non-limiting embodiment, the term is defined as being within 10% deviation, preferably within 5% deviation, more preferably within 1% deviation, and most preferably within 0.5% deviation. Additionally, "substantially non-aqueous" means less than 5%, 4%, 3%, 2%, 1% or less than 1% water by weight or volume.

[0051] A "pharmaceutically acceptable" ingredient, excipient or component is an ingredient, excipient or component that is suitable for use in humans and / or animals, has no excessive adverse side effects (such as toxicity, irritation and allergic reactions), and has a reasonable benefit / risk ratio.

[0052] A "pharmaceutically acceptable carrier" refers to a pharmaceutically acceptable solvent, suspending agent or vehicle for delivering the pharmaceutical compounds of the present invention to a mammal such as a human.

[0053] As used herein, an "ionization-stabilizing excipient" is an excipient that establishes and / or maintains a specific ionization state for a therapeutic agent. In certain aspects, an ionization-stabilizing excipient can be or include a molecule that provides at least one cation, particularly at least one divalent cation, under appropriate conditions, or a molecule that is a cation (particularly a divalent cation) donor compound or cation source.

[0054] As used herein, a "mineral acid" is an acid derived from one or more inorganic compounds. Thus, a mineral acid may also be referred to as an "inorganic acid". A mineral acid can be monoprotic or polyprotic (e.g., diprotic, triprotic, etc.). Non-limiting examples of mineral acids include hydrochloric acid (HCl), nitric acid (HNO 3 ), sulfuric acid (H 2 SO 4 ), and phosphoric acid (H 3 PO 4 ).

[0055] As used herein, a "mineral base" (which can equivalently or alternatively be referred to as an "inorganic base") is a basic compound derived from one or more inorganic substances. Many but not all inorganic bases are typically classified as "strong bases", and non-limiting examples of inorganic bases include sodium hydroxide (NaOH), potassium hydroxide (KOH), magnesium hydroxide (Mg(OH) 2 ), and calcium hydroxide (Ca(OH) 2 ).

[0056] As used herein, an "organic acid" is an organic compound that has acidity (i.e., can act as a proton source). Carboxylic acids, such as acetic acid or citric acid, are an example of organic acids. Other 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.).

[0057] As used herein, an "organic base" is an organic compound that has basicity (i.e., can act as a proton acceptor / proton sink). Many but not all organic bases contain nitrogen atoms (e.g., amines), and non-limiting examples of organic bases include amino acids (e.g., histidine, arginine, lysine), pyridine, imidazole, and tromethamine. Each molecule of an organic base can accept one or more protons.

[0058] "Charge distribution", "charge state", "ionization", "ionization state", and "ionization profile" are used interchangeably and refer to the ionization state resulting from the protonation and / or deprotonation of the ionizable groups of a peptide.

[0059] As used herein, a "composite formulation" is a formulation comprising two or more therapeutic agents dissolved in an aprotic polar solvent system. The therapeutic agents may belong to the same class (e.g., a composite formulation comprising two or more therapeutic peptides, such as insulin and pramlintide, or glucagon and GLP-1), or the therapeutic agents may belong to different classes (e.g., a composite formulation comprising one or more than one therapeutic small molecule and one or more than one therapeutic peptide molecule, such as GLP-1 and lisofylline).

[0060] When used in conjunction with the term "comprising" in the claims and / or description, the quantifier preceding the element may mean "a", and also mean "one or more", "at least one", and "one or more than one".

[0061] The terms "comprising", "having", "including" or "containing" are inclusive or open-ended and do not exclude additional, unrecited elements or method steps.

[0062] Other objects, features and advantages of the present invention will become apparent through the following specific embodiments. However, it should be understood that the specific embodiments and examples, although describing specific embodiments of the present invention, are given by way of illustration only. In addition, it is expected that changes and modifications within the spirit and scope of the present invention will become apparent to those skilled in the art from the specific embodiments. BRIEF DESCRIPTION OF THE DRAWINGS

[0063] The following drawings constitute part of this specification and are included to further illustrate certain aspects of the present invention. The invention may be better understood by reference to one or more of these drawings in conjunction with the detailed description of the embodiments of the specification provided herein.

[0064] Figures 1A to 1B is a pair of line graphs showing the mean plasma levothyroxine concentrations of minipigs administered various formulations of the present invention, on a linear scale ( Figure 1A ) or logarithmic scale ( Figure 1B )draw.

[0065] Figure 2 is a box plot showing the mean Tmax values ​​(in hours) for levothyroxine formulations of the invention administered subcutaneously to minipigs.

[0066] Figure 3 is a box plot showing mean Cmax values ​​(ng / mL) of the levothyroxine formulations of the present invention administered subcutaneously to minipigs.

[0067] Figure 4 is a box plot showing mean AUClast (hours*ng / ml) of levothyroxine formulations of the present invention administered subcutaneously to minipigs.

[0068] Figure 5 It is a box plot showing the mean AUC144 - 336 (hour * ng / ml) of the levothyroxine preparation of the present invention administered subcutaneously to minipigs.

[0069] Figures 6A to 6B It is a pair of line graphs showing the mean baseline unadjusted plasma levothyroxine concentrations in minipigs administered the 5 mg / mL levothyroxine preparation or the 10 mg / mL levothyroxine preparation (XP - 8121 - 5 and XP - 8121 - 10 respectively) of the present invention, plotted on a linear scale ( Figure 6A ) or a logarithmic scale ( Figure 6B ).

[0070] Figures 7A to 7B It is a pair of line graphs showing the mean baseline adjusted plasma levothyroxine concentrations in minipigs administered the 5 mg / mL levothyroxine preparation or the 10 mg / mL levothyroxine preparation (XP - 8121 - 5 and XP - 8121 - 10 respectively) of the present invention, plotted on a linear scale ( Figure 7A ) or a logarithmic scale ( Figure 7B ).

[0071] Figure 8 It is a line graph showing the mean baseline adjusted plasma diazepam concentrations (ng / mL) in minipigs administered the 50 mg / mL diazepam preparation or the 100 mg / mL diazepam preparation of the present invention, plotted on a linear scale.

[0072] Figure 9 It is a graphical description for a human clinical trial study design, named XP - 8121 - 108, to determine the pharmacokinetics of certain levothyroxine - containing preparations of the present invention administered subcutaneously (SC) relative to oral (PO) administration of tablet formulations .

[0073] Figures 10A to 10B It is a pair of line graphs showing the mean baseline adjusted plasma levothyroxine concentrations in human subjects in the XP - 8121 - 108 study following subcutaneous (SC) administration of the 10 mg / mL levothyroxine preparation (XP - 8121) of the present invention at 600 μg, 1200 μg, or 1500 μg compared to oral (PO) administration at 600 μg . The results are plotted on a linear scale ( Figure 10A ) or a semi - logarithmic scale ( Figure 10B ).

[0074] Figures 11A to 11B : In the XP - 8121 - 108 clinical study for subcutaneous administration of the levothyroxine preparation of the present invention to human subjects, Figure 11Ais a box plot showing the baseline-adjusted mean Tmax (in hours), Figure 11B is a bar graph of the baseline-adjusted mean half-life (T 1 / 2 , in hours).

[0075] Figures 12A to 12B is a pair of bar graphs showing the baseline-adjusted mean Cmax (ng / mL) ( Figure 12A ) or baseline-adjusted mean AUClast (ng*hour / mL) following subcutaneous administration of the levothyroxine formulation of the present invention to human subjects in the XP-8121-108 clinical study ( Figure 12B ).

[0076] Figures 13A to 13B is a pair of line graphs showing the baseline-adjusted mean Cmax normalized to the levothyroxine dose in human subjects who received subcutaneous (SC) administration of the 10 mg / mL levothyroxine formulation of the present invention (XP-8121) at 600 μg, 1200 μg, or 1500 μg in the XP-8121-108 study. Results are plotted on a linear scale ( Figure 13A ) or a logarithmic scale ( Figure 13B ).

[0077] Figures 14A to 14B is a pair of line graphs showing the baseline-adjusted mean AUClast normalized to the levothyroxine dose in human subjects who received subcutaneous (SC) administration of the 10 mg / mL levothyroxine formulation of the present invention (XP-8121) at 600 μg, 1200 μg, or 1500 μg in the XP-8121-108 study. Results are plotted on a linear scale ( Figure 14A ) or a logarithmic scale ( Figure 14B ). DETAILED DESCRIPTION

[0078] When formulated as an aqueous solution, standard small molecules, peptides, and protein molecules can be vulnerable to a variety of physical and chemical degradation pathways. For many of these therapeutic molecules, degradation pathways catalyzed, mediated, and / or facilitated by water (such as hydrolysis, racemization, deamidation) cannot be avoided, and thus the molecules cannot be adequately stabilized. As a result, many therapeutic agents cannot be formulated as stable solutions for parenteral injection and are instead formulated as powders that are reconstituted immediately before use.

[0079] To address the physical and / or chemical instability exhibited by many therapeutic molecules in water, formulations can be prepared in which the therapeutic agent is dissolved in a biocompatible non-aqueous liquid such as an aprotic polar solvent (e.g., DMSO). Prior non-aqueous formulations were based at least in part on the premise that restricting the water content of the formulation promotes physical and chemical stability by inhibiting water-mediated degradation pathways. Many of these known formulations limit the water content to a maximum of 10 weight / weight %.

[0080] Preparing non-aqueous therapeutic formulations using aprotic polar solvents to inhibit many common degradation pathways, particularly those involving water, can significantly improve the stability of one or more soluble or dissolved therapeutic molecules. However, problems still exist with the compositions and methods disclosed in the art. In particular, directly dissolving a therapeutic molecule in an aprotic polar solvent is not a suitable method for preparing stable compositions of most therapeutic molecules. When directly dissolved in DMSO, various therapeutic agents, such as levothyroxine at a concentration of 5 mg / mL, discolor and form degradation products within a day even when refrigerated. For a composition containing only levothyroxine and DMSO, 5 mg / mL corresponds to approximately 0.45 weight / weight % of levothyroxine, indicating that even at relatively low concentrations, direct dissolution in an aprotic polar solvent system itself does not prevent degradation and / or gelation of the therapeutic molecule. In addition, therapeutic molecules that are not readily chemically degraded in an aprotic polar solvent system may still form insoluble aggregates when directly dissolved in the aprotic polar solvent system.

[0081] Without wishing to be bound by theory, it is believed that for a therapeutic molecule to exhibit enhanced or optimal stability and solubility when formulated in an aprotic polar solvent system, the therapeutic molecule may require a specific ionization profile. An ionization profile is the charge state obtained by protonation and / or deprotonation of the ionizable groups of a therapeutic molecule, such that the molecule has an overall positive charge state, negative charge state, or neutral charge state. For example, protonation of ionizable amino acid residues (e.g., arginine, lysine, aspartic acid, glutamic acid) contained in a therapeutic peptide can confer an overall positive charge on the molecule in solution. Alternatively, deprotonation of ionizable amino acid residues can render the molecule in solution overall negatively charged. Although deprotonation of ionizable amino acid residues in therapeutic peptide molecules is also contemplated within the scope of the present invention, for the non-limiting examples used herein, protonated (i.e., positively charged) molecules will be described. In such embodiments, relatively long-range electrostatic repulsion between positively charged API molecules can inhibit shorter-range hydrophobic interactions that lead to physical instabilities (e.g., precipitation, aggregation, and / or gelation). Thus, in the absence of sufficient protonation (i.e., an optimal or beneficial ionization profile), a therapeutic molecule dissolved in an aprotic polar solvent system can be physically unstable and result in the formation of soluble and / or insoluble aggregates or other degradation products. Accordingly, it may be necessary to include at least one excipient present in a concentration sufficient to act as an ionization stabilizer, which is capable of conferring an ionization profile on the active agent in the aprotic polar solvent system that improves physical stability and / or chemical stability. The appropriate concentration of one or more ionization-stabilizing excipients to be added to the solution depends on several factors, including but not limited to the chemical structure of the ionization-stabilizing excipient, the chemical structure of the one or more active agents, the concentration of the one or more active substances, the solvent system used, the presence of co-solvents, and the presence and respective concentrations of other excipients or formulation components.

[0082] Certain compositions and methods have been designed to establish an optimal ionization profile for a therapeutic molecule prior to its dissolution in an aprotic polar solvent system. For example, a peptide powder from a vendor / manufacturer is first dissolved in a buffered aqueous solution, wherein the pH of the buffered aqueous solution of the peptide is set to the optimal stability and solubility for the particular peptide. The peptide is then dried (e.g., by lyophilization or spray drying) from the aqueous solution into a powder such that the ionization profile of the peptide molecules in the powder can be approximately equal to the ionization profile of the peptide molecules dried from the aqueous solution. Then when the peptide powder is dissolved in an aprotic polar solvent system, the ionization profile of the peptide molecules can be approximately equal to the ionization profile of the peptide molecules in the powder. Thus, the ionization profile of the peptide molecules in the aprotic polar solvent system is approximately equal to the ionization profile of the peptide molecules in the buffered aqueous solution (see U.S. Pat. Nos. 9,649,364, 10,485,850, and 11,020,403, the disclosures of which are incorporated herein by reference in their entireties).

[0083] In order to optimize the ionization profile of a therapeutic molecule and confer pH memory prior to dissolution in a non-protic polar solvent, it is necessary to dry the therapeutic molecule from an aqueous buffer solution, which typically adds a significant cost to the formulation development pathway in terms of time and expense. In particular, it is well known that the drying process imposes several stresses on the therapeutic molecule, and a sufficient amount of additional excipients (e.g., lyoprotectants such as trehalose and sucrose, and / or surfactants such as polysorbate 80) must be added to the aqueous solution to protect the therapeutic molecule, thereby increasing the cost and complexity of the formulation. In addition, the drying process (e.g., spray drying, freeze drying) typically must be optimized for a given therapeutic molecule both at the laboratory scale during the initial development of the process and at the production scale as the process is scaled up and transferred to an instrument and facility capable of producing commercial scale batches. Thus, the initial development and optimization of the drying process for a given therapeutic molecule, along with the associated time and cost of transferring the method and integrating other steps in the manufacturing process can be very expensive. Without wishing to be bound by theory, it is believed that by providing a sufficient amount of at least one ionization-stabilizing excipient to achieve an appropriate or optimal ionization profile of the therapeutic molecule, the electrostatic repulsion between therapeutic molecules having the same charge polarity (i.e., negatively charged or positively charged) can be sufficient in magnitude to prevent physical degradation (e.g., by short-range hydrophobic interactions between molecules that cause aggregation). This is particularly important for molecules that exhibit a tendency to aggregate in solution, especially when the concentration of the molecules in the solution increases. In addition, by controlling and optimizing the degree of ionization (i.e., protonation or deprotonation) of the therapeutic agent, chemical degradation can be minimized. For example, excessive protonation can promote chemical instability through degradation reactions such as oxidation (e.g., oxidation of methionine residues) and fragmentation (e.g., cleavage of the peptide backbone). Thus, for some therapeutic molecules, there may be an optimal or beneficial ionization profile achieved through protonation or deprotonation, thereby minimizing physical and / or chemical degradation reactions. For small molecule or peptide therapeutics, the degree of ionization (i.e., protonation or deprotonation) required for stability and thus the amount of ionization-stabilizing excipient required in solution will depend on the structure of the therapeutic molecule and its concentration in solution.

[0084] Each molecule used as an ionization stabilizing excipient exhibits some tendency to donate or accept a proton to / from one or more therapeutic molecules and / or additional pharmaceutical substances / powder components (such as salts, counterions, buffer molecules, etc.) in a given solvent system; the tendency to donate a proton can be referred to as the relative acid strength of the molecule, and the tendency to accept a proton can be referred to as the relative base strength of the molecule. As a non-limiting example, for a fixed concentration of a proton donor molecule (for simplicity, only a monoprotic molecule is assumed in this example), a molecule with greater acid strength will protonate the therapeutic molecule to a greater extent than a weaker acid. Thus, the concentration of a given proton donor molecule (ionization stabilizing excipient) required to achieve an appropriate or optimal ionization profile for a therapeutic molecule will be inversely proportional to its acid strength. These and other non-limiting aspects of the invention are disclosed herein.

[0085] The formulation may contain an ionizing stabilizing excipient at a concentration of at least or about 0.01 mM, 0.1 mM, 0.5 mM, 1 mM, 10 mM, 15 mM, 20 mM, 25 mM, 30 mM, 35 mM, 40 mM, 45 mM, or 50 mM and all ranges and all concentrations therebetween up to at most or about 10 mM, 15 mM, 20 mM, 25 mM, 30 mM, 50 mM, 75 mM, 100 mM, 500 mM, or 1000 mM and all ranges and all concentrations therebetween, or up to the solubility limit of the ionizing stabilizing excipient in the aprotic polar solvent system. The appropriate amount of the ionizing stabilizing excipient to include can be readily determined based on the charge distribution of the therapeutic agent being dissolved in the aprotic polar system. In particular, the optimal amount of the ionizing stabilizing excipient added in the aprotic polar solvent is an ionizing stabilizing excipient sufficient to produce from about 2x to 3x moles of acid per mole of therapeutic agent (e.g., about 14 mM acid for a 5 mg / mL (about 6.4 mM) sodium levothyroxine solution; about 10 mM acid for a 5 mg / mL (about 5 mM) free levothyroxine solution), and the concentration of the API in the final formulation, i.e., the molar ratio of the ionizing stabilizing excipient to the therapeutic agent, is from about 2:1 to about 3:1, and preferably about 2.5:1. In certain aspects, the ionizing stabilizing excipient concentration is from about 0.1 mM to about 100 mM, particularly from about 1 mM to about 35 mM, e.g., about 1 mM, about 2 mM, about 5 mM, about 10 mM, about 15 mM, about 20 mM, about 25 mM, about 30 mM, and about 35 mM and all ranges and all concentrations therebetween. Certain such aspects include an ionizing stabilizing excipient at a concentration selected from about 15 mM to about 32 mM, including about 15 mM, about 16 mM, about 17 mM, about 18 mM, about 19 mM, about 20 mM, about 21 mM, about 22 mM, about 23 mM, about 24 mM, about 25 mM, about 26 mM, about 27 mM, about 28 mM, about 29 mM, about 30 mM, about 31 mM, and about 32 mM and all ranges and all concentrations therebetween. In certain such aspects, the formulation of the invention contains an ionizing stabilizing excipient at a concentration of from about 26 mM to about 32 mM, particularly about 26 mM, about 27 mM, about 28 mM, about 29 mM, about 30 mM, about 31 mM, and about 32 mM and all ranges and all concentrations therebetween. In certain embodiments, the ionizing stabilizing excipient can be a suitable mineral acid, such as hydrochloric acid, sulfuric acid, nitric acid, phosphoric acid, etc. Particularly preferred mineral acids are hydrochloric acid and sulfuric acid. In certain aspects, the ionizing stabilizing excipient can be an organic acid, e.g., an amino acid, an amino acid derivative, or a salt of an amino acid or an amino acid derivative (examples include glycine, trimethylglycine (betaine), glycine hydrochloride, and trimethylglycine (betaine) hydrochloride). In other aspects, the amino acid can be glycine or the amino acid derivative trimethylglycine.

[0086] In some aspects, an aprotic polar solvent can be deoxygenated prior to preparing a formulation. Many different techniques can be used in the context of the present invention to deoxygenate or remove oxygen (degas or deoxygenate) from an aprotic polar solvent. For example, it is contemplated that deoxygenation can, but is not limited to, removing dissolved oxygen in a liquid aprotic polar solvent by the liquid alone, by the liquid and other solute molecules (such as micelles, cyclodextrins, etc.), or by other solute molecules alone. Non-limiting examples of deoxygenation techniques include subjecting the aprotic polar solvent to reduced pressure and / or heating the liquid to reduce the solubility of dissolved gases, fractional distillation, membrane degassing, inert gas displacement, using reducing agents, freeze-pump-thaw cycles, or storing for an extended period in a gas-lock container. In one embodiment, the aprotic polar solvent is deoxygenated by vacuum degassing. In another embodiment, the aprotic polar solvent is deoxygenated by using a deaerator. In one case, the deaerator is a tray-type or cascade-type deaerator. In another case, the deaerator is a spray-type deaerator. In yet another embodiment, the aprotic polar solvent is deoxygenated using a gas-liquid separation membrane. In one case, the aprotic polar solvent is degassed using a gas-liquid separation membrane and reduced pressure. In one embodiment, a non-oxygen gas (such as N 2 ) is bubbled through the liquid to replace or reduce the oxygen in the aprotic polar solvent. In one case, the gas bubbled through the aprotic polar solvent is argon, helium, nitrogen, an inert gas, and / or hydrogen, preferably nitrogen. In another case, a degassing tower is used to bubble the gas through the aprotic polar solvent. In yet another embodiment, the aprotic polar solvent is deoxygenated by one or more reducing agents. Non-limiting examples of reducing agents include ammonium sulfite, hydrogen, active deoxygenating metals, copper, tin, cadmium, Wood's metal alloy (50% bismuth, 25% lead, 12.5% tin, and 12.5% cadmium), etc. In yet another embodiment, the aprotic polar solvent is degassed by a freeze-pump-thaw cycle (e.g., at least 1, 2, 3, or more than 3 cycles can be used). In one case, the freeze-pump-thaw cycle includes freezing the aprotic polar solvent under liquid nitrogen, applying a vacuum, and then thawing the solvent in warm water. In one embodiment, the aprotic polar solvent is deoxygenated by long-term storage in a steel container, a glass container, or a wooden container. In another embodiment, the aprotic polar solvent is sonicated, ultrasonically treated, or stirred during the deoxygenation process.

[0087] After being treated or deoxygenated, the aprotic polar solvent can have a dissolved oxygen of less than 0.1 mM, preferably less than 0.05 mM. Methods known to those skilled in the art can be used to determine the amount of dissolved oxygen in any given aprotic polar solvent (e.g., a dissolved oxygen meter or probe device can be used, such as a dissolved oxygen probe commercially available from Vernier (Beaverton, Oregon, USA)).

[0088] In some aspects, the formulations disclosed in the present application can be prepared and / or sealed under an inert gas atmosphere. Common methods include backfilling a primary container closure system (e.g., vial) to provide an inert gas (e.g., nitrogen, argon) headspace. A secondary container closure system (e.g., a sealed foil bag) can also be sealed under an inert gas environment.

[0089] I. Formulations

[0090] The formulations of the present invention comprise a therapeutic agent present in an aprotic polar solvent system containing at least one ionization-stabilizing excipient and optionally one or more additional pharmaceutically acceptable excipients. The therapeutic agent can be dissolved (e.g., completely or partially) or suspended (completely or partially) in the aprotic polar solvent system.

[0091] In some embodiments, the therapeutic agent is present in an aprotic polar solvent that further comprises one or more ionization-stabilizing excipients, such as those described above and in the concentrations described above. In other embodiments, the therapeutic agent is present in an aprotic polar solvent that is "pure", i.e., does not contain a co-solvent, or, if it contains a co-solvent, does not contain a co-solvent other than water. In other embodiments, the therapeutic agent is present in a solvent system that is a mixture of two or more aprotic polar solvents and water or moisture in an amount greater than 10 volume / volume% (i.e., an aprotic polar solvent system). For example, a solvent system comprising a 75 / 25 volume / volume% mixture of DMSO and NMP with a total water content greater than 10 volume / volume%. However, in some embodiments, a co-solvent can be used, wherein the co-solvent is mixed in one or more aprotic polar solvents. Non-limiting examples of co-solvents (explicitly excluding water) include ethanol, propylene glycol (PG), glycerol, and mixtures thereof. The co-solvent can be present in the formulation in an amount of from about 0.1 weight / volume% to about 50 weight / volume%, such as about 0.1 weight / volume%, about 0.5 weight / volume%, about 1 weight / volume%, about 5 weight / volume%, about 10 weight / volume%, about 15 weight / volume%, about 20 weight / volume%, about 25 weight / volume%, about 30 weight / volume%, about 35 weight / volume%, or about 40 weight / volume%. In some embodiments, the co-solvent is present in an amount of from about 10 weight / volume% to about 50 weight / volume%, from about 10 weight / volume% to about 40 weight / volume%, from about 10 weight / volume% to about 30 weight / volume%, from about 10 weight / volume% to about 25 weight / volume%, from about 15 weight / volume% to about 50 weight / volume%, from about 15 weight / volume% to about 40 weight / volume%, from about 15 weight / volume% to about 30 weight / volume%, or from about 15 weight / volume% to about 25 weight / volume%.

[0092] Certain preferred but non-limiting exemplary formulations of the present invention for treating or preventing hypothyroidism, or diseases or disorders associated with or caused thereby, comprising levothyroxine at a concentration of about 5 mg / mL or about 10 mg / mL, dissolved in a solution comprising DMSO and an ionization stabilizing component, such as sulfuric acid at a concentration of about 13 mM to about 35 mM, such as about 26 mM to about 32 mM and all ranges and all concentrations therebetween.

[0093] In addition, the formulations of the present invention may further comprise one or more other excipients in addition to at least one ionization-stabilizing excipient. In some embodiments, the other excipients are selected from the group consisting of sugars, salts, starches, sugar alcohols, antioxidants, chelating agents, and preservatives. Examples of suitable sugar excipients include, but are not limited to, trehalose, glucose, sucrose, etc. Examples of suitable starches for stabilizing excipients include, but are not limited to, hydroxyethyl starch (HES). Examples of suitable sugar alcohols (also known as polyols) for stabilizing excipients include, but are not limited to, mannitol and sorbitol. Examples of suitable antioxidants include, but are not limited to, ascorbic acid, cysteine, methionine, thioglycerol, 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 (sodium ethylenediaminetetraacetate), tartaric acid and its salts, glycerol, and citric acid and its salts. Examples of suitable inorganic salts include, but are not limited to, sodium chloride, potassium chloride, calcium chloride, magnesium chloride, calcium sulfate, magnesium sulfate, zinc sulfate, and zinc acetate. Examples of suitable preservatives include, but are not limited to, benzyl alcohol, methylparaben, m-cresol, propylparaben, and mixtures thereof. Other formulation components include local anesthetics such as lidocaine or procaine. In some embodiments, the other stabilizing excipients are present in the formulation in an amount of from about 0.01% weight / volume to about 60% weight / volume, from about 1% weight / volume to about 50% weight / volume, from about 1% weight / volume to about 40% weight / volume, from about 1% weight / volume to about 30% weight / volume, from about 1% weight / volume to about 20% weight / volume, from about 5% weight / volume to about 60% weight / volume, from about 5% weight / volume to about 50% weight / volume, from about 5% weight / volume to about 40% weight / volume, from about 5% weight / volume to about 30% weight / volume, from about 5% weight / volume to about 20% weight / volume, from about 10% weight / volume to about 60% weight / volume, from about 10% weight / volume to about 50% weight / volume, from about 10% weight / volume to about 40% weight / volume, from about 10% weight / volume to about 30% weight / volume, or from about 10% weight / volume to about 20% weight / volume.In some embodiments, other stabilizing excipients are present in the formulation in an amount of about, at most, or at least 0.01% weight / volume, 0.1% weight / volume, 0.5% weight / volume, 1% weight / volume, 2% weight / volume, 3% weight / volume, 4% weight / volume, 5% weight / volume, 6% weight / volume, 7% weight / volume, 8% weight / volume, 9% weight / volume, 10% weight / volume, 15% weight / volume, 20% weight / volume, 25% weight / volume, 30% weight / volume, 35% weight / volume, 40% weight / volume, 45% weight / volume, 50% weight / volume, 55% weight / volume, or 60% weight / volume.

[0094] II. Therapeutic Agent

[0095] Therapeutic agents in the context of the present invention include peptide or protein compounds, small molecule drugs, and pharmaceutically acceptable analogs and / or salts thereof. One of ordinary skill in the art knows which therapeutic agent is suitable for treating certain diseases or disorders and is able to administer an effective amount of the therapeutic agent in the formulation as described herein to treat the disease or disorder.

[0096] Non-limiting examples of small molecule drugs (and their salts) that can be used in the context of the present invention include, but are not limited to, levothyroxine, epinephrine, benzodiazepines Drugs, catecholamines, "triptans", sumatriptan, nosoline, small molecule chemotherapeutics (such as mitoxantrone), small molecule corticosteroids (such as methylprednisolone, beclomethasone dipropionate), small molecule immunosuppressants (such as azathioprine, cladribine, cyclophosphamide monohydrate, methotrexate), small molecule anti-inflammatories (such as salicylic acid, acetylsalicylic acid, lysophylline, diflunisal, choline magnesium trisalicylate, salicylates, benorilate, flufenamic acid, mefenamic acid, meclofenamic acid, triflumic acid, diclofenac, fenclofenac, alclofenac, fenclozic acid, ibuprofen, flurbiprofen, ketoprofen, naproxen, fenoprofen, fenbufen, sulprofen, indoprofen, tiaprofenic acid, benoxaprofen, pirprofen, tolmetin, zomepirac, clopinac, indomethacin, sulindac, phenylbutazone, oxyphenbutazone, azapropazone, metamizole, piroxicam, isoxicam), small molecules for treating neurological diseases (such as cimetidine, ranitidine, famotidine, nizatidine, tacrine, metrifonate, galantamine, selegiline, imipramine, fluoxetine, olanzapine, sertindole, risperidone, sodium valproate semisodium, gabapentin, carbamazepine, topiramate, phenytoin), small molecules for treating cancer (such as vincristine, vinblastine, paclitaxel, docetaxel, cisplatin, irinotecan, topotecan, gemcitabine, temozolomide, imatinib, bortezomib), statins (such as atorvastatin, amlodipine, rosuvastatin, sitagliptin, simvastatin, fluvastatin, pitavastatin, lovastatin, pravastatin, simvastatin) and other taxane derivatives, small molecules for treating tuberculosis (such as rifampicin), small molecule antifungals (such as fluconazole), small molecule anxiolytics and small molecule anticonvulsants (such as lorazepam), small molecule anticholinergics (such as atropine), small molecule β-agonist drugs (such as salbutamol sulfate), small molecule mast cell stabilizers and small molecule drugs for treating allergies (such as sodium cromoglycate), small molecule anesthetics and small molecule antiarrhythmics (such as lidocaine), small molecule antibiotics (such as tobramycin, ciprofloxacin), small molecule antimigraine drugs (such as sumatriptan) and small molecule antihistamines (such as diphenhydramine). In a preferred embodiment, the small molecule is levothyroxine. Based on the information provided herein and readily available in the art, those skilled in the art will generally be familiar with other suitable examples of such small molecule drugs (and their salts) that can be advantageously used in the compositions and methods of the present invention.

[0097] Non-limiting examples of peptides and proteins (and their salts) that can be used in the context of the present invention include, but are not limited to, glucagon, pramlintide, insulin, leuprolide, luteinizing hormone releasing hormone (LHRH) agonists, adrenocorticotropic hormone (ACTH), leuprolide, hirudin, parathyroid hormone (PTH), dextrin, angiotensin (1-7), botulinum toxin, hematide, amyloid peptides, gastric inhibitory polypeptide, antibodies (which can be monoclonal or polyclonal) or fragments thereof, immunogenic peptides (e.g., peptides or peptide complexes derived from viruses, bacteria, or any prokaryote or any eukaryote or their cells), insulin-like growth factors, growth hormone releasing factor, antimicrobial factors, glatiramer acetate, glucagon-like peptide-1 (GLP-1), GLP-1 agonists, exenatide, its analogs, dextrin analogs (pramlintide), and mixtures thereof. In some preferred aspects, the therapeutic agent is glucagon, insulin, and / or pramlintide. Based on the information provided herein and readily available in the art, those of ordinary skill in the art will generally be familiar with other suitable examples of such peptides, proteins, peptide complexes, and their derivatives that can be advantageously used in the compositions and methods of the present invention.

[0098] Each of the above-mentioned small molecule drugs, peptides, and proteins is well-known and commercially available from various manufacturers and sources. In addition, the amounts of small molecule drugs, peptides, and proteins in the dosage formulations provided by the present invention can vary according to currently acceptable amounts, the needs of the subject / patient (e.g., age, health status, weight, nature and extent of symptoms), etc.; such amounts can be readily determined by those of ordinary skill in the pharmaceutical and pharmacological arts based on readily available information.

[0099] Therapeutic agents provided by manufacturers or commercial sources are typically provided in powder form for dissolution into the formulations as described herein. Many known techniques can be used to form the powder for dissolution.

[0100] Any suitable dosage of small molecule drugs, peptides, and proteins (each being an "active pharmaceutical ingredient" or "API") can be formulated in the stable formulations of the present invention. Generally, the API (or each API in embodiments comprising two or more APIs) is present in the formulation in an amount from about 0.1 μg / mL to up to the solubility limit of the API. In some such embodiments, the dosage is from about 0.1 μg / mL to about 500 mg / mL, or up to about 1 mg / mL, about 2.5 mg / mL, about 5 mg / mL, about 7.5 mg / mL, about 10 mg / mL, about 15 mg / mL, about 20 mg / mL, about 25 mg / mL, about 50 mg / mL, about 75 mg / mL, about 100 mg / mL, about 150 mg / mL, about 200 mg / mL, about 250 mg / mL, about 300 mg / mL, about 350 mg / mL, about 400 mg / mL, about 450 mg / mL, or about 500 mg / mL and all dosages between the two ends of each range described herein. In some embodiments, the small molecule API is levothyroxine, which is present in the formulation in an amount from about 2 mg / mL to about 40 mg / mL, advantageously at about 5 mg / mL, about 10 mg / mL, about 15 mg / mL, about 20 mg / mL, or about 40 mg / mL. In other embodiments, levothyroxine is present in an amount from about 3 mg / mL to about 20 mg / mL, and preferably at 5 mg / mL or 10 mg / mL. In some embodiments of the present invention, for subcutaneous or intramuscular injection in humans, a volume sufficient to deliver from about 500 μg to about 1500 μg of levothyroxine and all amounts within this range of the formulation are introduced into a subject, such as a human or veterinary animal. In some such embodiments, levothyroxine is injected into the subject in a volume suitable to deliver about 600 μg, about 1200 μg, or about 1500 μg of levothyroxine per injection. Similarly, based on the information provided herein and readily available in the relevant art, it will be apparent to those skilled in the art that the dosage can vary depending on the active ingredient used and the disease, disorder, or condition to be treated.

[0101] In some embodiments, the formulations of the present invention further comprise an antioxidant. In other embodiments, the formulation further comprises a chelating agent. In still other embodiments, the formulations of the present invention further contain a preservative, a sugar (such as a monosaccharide, disaccharide, or polysaccharide, such as trehalose dihydrate), a sugar alcohol (such as mannitol, xylitol, or erythritol), a polyol, a surfactant, and / or a salt.

[0102] III. Method of treatment

[0103] In another aspect, the present invention provides a method for treating or preventing a disease, condition or disorder by administering a therapeutically effective amount of a therapeutic agent for treating or preventing a disease, condition or disorder in the formulations described herein to a subject. The therapeutic agent of the present invention can be administered intradermally, preferably subcutaneously or intradermally, and most preferably subcutaneously, in the prevention, diagnosis, alleviation, treatment or cure of diseases or physical conditions in humans and veterinary animals.

[0104] In some embodiments, the methods of treatment of the present invention include treating hypothyroidism or a disease or physical condition arising from or associated with hypothyroidism by administering to a subject having hypothyroidism or a disease or physical condition associated therewith an effective amount of a hypothyroidism therapeutic agent that treats hypothyroidism or a disease or physical condition arising from or associated with hypothyroidism, preferably levothyroxine. The compositions and methods of the present invention are advantageously used to treat hypothyroidism regardless of its etiology, i.e., whether it results from a primary or secondary cause. Any condition that affects the thyroid and results in its production of low levels of thyroid hormones is considered a primary cause of hypothyroidism, while secondary causes are any conditions, disorders or injuries that result in pituitary failure, causing it to be unable to secrete thyroid stimulating hormone (TSH), which signals the thyroid to secrete a greater amount of thyroid hormones to overcome the imbalance, which is characteristic of hypothyroidism. Such diseases or conditions associated with or attributable to hypothyroidism include, but are not limited to, thyroiditis, congenital hypothyroidism, autoimmune hypothyroidism (such as Hashimoto's disease), ablation of the thyroid by surgical, radiological or chemical means (e.g., in the treatment of hyperthyroidism or its effects, such as Grave's disease or thyroid eye disease), iodine deficiency, myxedema, myxedema coma / crisis, postpartum thyroiditis, certain viral diseases, etc. Patients with hypothyroidism report a number of symptoms, not all of which are associated with every case of hypothyroidism. These symptoms include, for example, fatigue, peripheral neuropathy (especially numbness or tingling sensations in the hands and / or fingers), weight gain, development of goiter, fibromyalgia, body aches including muscle weakness or joint pain, hypercholesterolemia, depression, low body temperature or increased sensitivity to cold, dry / rough skin and hair, hair loss, physical changes in facial appearance (e.g., drooping eyelids or puffiness of the eyes / facial area), cognitive problems or memory problems (often described as "brain fog"), etc. By treating and / or preventing hypothyroidism, the levothyroxine-containing compositions and methods of the present invention can improve or restore the thyroid hormone balance in the subject and thus alleviate or reduce the above symptoms. In the non-limiting specific exemplary embodiments described in the examples below, the formulations of the present invention can be administered to the subject by subcutaneous administration, overcoming the limitations of conventional daily oral (per os or "PO") levothyroxine solid dosage forms such as tablets, by allowing once-weekly, once-every-two-months or even once-monthly administration of levothyroxine to the subject, thereby improving the patient experience and increasing their compliance with the treatment regimen established and prescribed by the patient's medical practitioner or other healthcare provider.

[0105] In some embodiments, the methods of treatment of the present invention include treating hypoglycemia by administering to a subject suffering from hypoglycemia an effective amount of a therapeutic agent for treating hypoglycemia in a storage-stable, sustained-release formulation as described herein. In some embodiments, a storage-stable, sustained-release formulation comprising glucagon is administered to the subject in a manner such that glucagon is released from the site of administration into the bloodstream or tissue of the animal over an extended period of time (i.e., sustained release) as compared to an immediate-release formulation comprising glucagon. In such aspects, the disease, condition, or disorder to be treated using the stable formulations of the present invention is a diabetic condition. Examples of diabetic conditions include, but are not limited to, type 1 diabetes, type 2 diabetes, gestational diabetes, prediabetes, hyperglycemia, hypoglycemia, and metabolic syndrome. In some embodiments, the disease, condition, or disorder is hypoglycemia, including but not limited to diabetes-related hypoglycemia, exercise-induced hypoglycemia, and post-bariatric surgery hypoglycemia, or other types of hypoglycemia known to those of ordinary skill in the art as described herein (see, e.g., U.S. Patent Nos. 9,649,364, 10,485,850, and 11,020,403, the disclosures of which are incorporated herein by reference in their entireties). In some embodiments, the disease, condition, or disorder is diabetes.

[0106] In some embodiments, the methods of treatment of the present invention include treating diabetes by administering to a patient having diabetes an effective amount of a therapeutic agent in a stable formulation as described herein for treating diabetes. In some embodiments, a stable formulation comprising insulin is administered to the subject. In some embodiments, a stable formulation comprising pramlintide is administered to the subject. In some embodiments, a stable formulation comprising insulin and pramlintide is administered to the subject. In some embodiments, a stable formulation comprising exenatide is administered to the subject. In some embodiments, a stable formulation comprising glucagon and exenatide is administered to the subject.

[0107] In certain aspects, the formulations of the present invention comprising epinephrine can be administered to a subject at risk of anaphylaxis or suspected of being at risk of anaphylaxis. Epinephrine is designated for the emergency treatment of type I anaphylactic reactions that can arise from a variety of sources, including but not limited to food, drugs, and / or other allergens, allergen immunotherapy, diagnostic test substances, insect stings and insect bites, and idiopathic or exercise-induced anaphylaxis.

[0108] The dosage of a small molecule drug or a peptide drug for treating a disease, condition or disorder (such as hypothyroidism, diabetic conditions, hypoglycemia or allergic reactions) as described herein is in accordance with the dosages and scheduling regimens practiced by those skilled in the art and as described herein. Goodman and Gilman's The Pharmacological Basis of Therapeutics, 11th Edition, 2006, supra, and Physicians' Desk Reference (PDR), e.g., in the 65th Edition (2011) or 66th Edition (2012), PDR Network, LLC, provide general guidelines for the appropriate dosages of all pharmaceutical agents used in the present invention, each of which is hereby incorporated herein by reference. The appropriate dosage of a small molecule drug or a peptide drug for treating a disease, condition or disorder as described herein will vary depending on a variety of factors including the formulation of the composition, the patient response, the severity of the condition, the weight of the subject and the judgment of the prescribing physician. The formulation of the effective dosage delivers a medically effective amount of the small molecule drug or the peptide drug. Adjustments to the dosage (i.e., increasing or decreasing over time) can be determined according to the needs of the individual patient or by a medical professional. Thus, those skilled in the art are fully capable of determining the effective amount or dosage of the small molecule drug or the polypeptide drug and thus determining the appropriate amount or volume of the therapeutic formulation of the present invention to be administered to a subject, particularly in accordance with the detailed disclosure provided herein.

[0109] The formulations of the present invention can be used for parenteral administration, including but not limited to subcutaneous administration, intradermal administration, intramuscular administration, intranasal administration, oral administration, transdermal administration or intravenous administration (e.g., by injection or infusion). In some embodiments, the formulation is administered subcutaneously. The formulation can also be delivered transdermally, e.g., by topically applying the composition to the skin (e.g., spreading the application over the skin or loading the composition in a skin patch and attaching the skin patch to the skin).

[0110] The formulations of the present disclosure can be administered by infusion or by injection using any suitable device. For example, the formulations of the present invention can be placed in a syringe (e.g., a pre-filled syringe), a pen injection device, an auto-injector device, or a pump device. In some embodiments, the injection device is a multi-dose syringe pump device or a multi-dose auto-injector device. The formulation is presented in the device in such a way that the formulation can easily flow out of the needle under the drive of the injection device, such as an auto-injector, to deliver the peptide drug. Suitable pen injection devices / auto-injector devices include, but are not limited to, those pen injection devices / auto-injector devices manufactured by Becton-Dickinson, Swedish Healthcare Limited (SHL Group), YpsoMed Ag, etc. Suitable pump devices include, but are not limited to, those pump devices manufactured by Tandem Diabetes Care, Inc., Delsys Pharmaceuticals, etc.

[0111] In some embodiments, there is provided a formulation of the present invention ready to be administered in a vial, an infusion device, or a pre-filled syringe.

[0112] In other embodiments, the formulations provided by the present invention can be used in certain diagnostic procedures. In particular, the glucagon-containing formulations of the present invention can be administered to a mammal such as a human or veterinary animal before one or more than one diagnostic procedure, as an adjunct to the diagnostic procedure, as part of the diagnostic procedure, or in combination with the diagnostic procedure, thereby providing a method for diagnosing a disease or disorder in a patient suffering from or susceptible to a disease or disorder. Non-limiting examples of diagnostic procedures in which the glucagon-containing formulations of the present invention can be appropriately used include methods for diagnosing Alzheimer's disease (see U.S. Patent No. 4,727,041, incorporated herein by reference in its entirety) and growth hormone deficiency (see U.S. Patent No. 5,065,747; also see Boguszewski, C.L., Endocrine 57:361-363 (2017), and Yuen, K.C.J., ISRN Endocrinology, vol. 211, Article ID 608056, pp. 1-6 (2011), doi:10.5402 / 2011 / 608056; the entire disclosures of which are incorporated herein by reference in their entirety). Other examples of such uses include in certain radiological diagnostic procedures, particularly those for diagnosing gastrointestinal disorders (non-limiting examples of which include abdominal obstruction, appendicitis, Barrett's esophagus, celiac disease, cancer, cirrhosis, Crohn's disease, diverticulitis, diverticulosis, ulcers, gallstones, gastric prolapse, gastritis, gastroesophageal reflux disease, hepatitis (A / B / C), hiatal hernia, inflammatory bowel disease, hernia, irritable bowel syndrome, pancreatitis, perianal fistula, ulcerative colitis, etc.), during radiological examination of the gastrointestinal system, temporarily inhibiting the motility of the gastrointestinal organs and connective tissues of an adult patient (see, for example, the label of the glucagon lyophilized product (NDC code 63323-185-03), available at: https: / / dailymed.nlm.nih.gov / dailymed / drugInfo.cfm?setid=8c8acad6-44cc-43aa-966b-027e053be8f5; also see Glucagon in Gastroenterology, J. Picazo, ed., Lancaster, England: MTP Press Ltd. (1979), particularly Chapters 3-7, pages 39-120; the disclosure of which is incorporated herein by reference in its entirety).In such diagnostic methods, the glucagon-containing preparation of the present invention can be administered to a patient suffering from or susceptible to the disorder by any means suitable for introducing such a preparation into the patient, such as the means described herein. For example, about 1 minute to 10 minutes before a diagnostic test (e.g., a radiological examination), it can be intravenously injected at a dose of about 0.2 mg to about 0.75 mg, or about 5 minutes to 15 minutes before a diagnostic test (e.g., a radiological examination), it can be intramuscularly or intradermally injected at a dose of about 1 mg to about 2 mg. Based on the content disclosed herein and the information available in the art, other suitable methods of therapeutic and diagnostic use of the preparation of the present invention will be readily familiar to a clinician or pharmacist of ordinary skill in the art.

[0113] IV. Kit / Container

[0114] Kits are also contemplated to be provided by certain aspects of the present invention. For example, the preparation of the present invention can be included in a kit, which can include a container. In one aspect, for example, the preparation can be contained in a container ready for administration to a subject or incorporated into a device configured to administer to a subject without reconstitution or dilution of the preparation. That is, the preparation to be administered can be stored in the container and readily used as needed. In some embodiments, the container can be a device. The device can be a syringe (e.g., a prefilled syringe), a pen injection device, an autoinjector device, a device capable of pumping or administering the preparation (e.g., an automatic or non-automatic external pump (e.g., a patch pump or a pump requiring an external infusion set), an implantable pump, etc.) or an infusion bag. Suitable pen injection devices / autoinjector devices include, but are not limited to, those manufactured by Becton-Dickinson, Swedish Healthcare Limited (SHL Group), YpsoMed Ag, etc. Suitable pump devices include, but are not limited to, those manufactured by Tandem Diabetes Care, Inc., Delsys Pharmaceuticals, etc. Suitable infusion sets include, but are not limited to, those produced / distributed / sold by companies such as Tandem Diabetes Care, Inc., Medtronic, Disetronic, YpsoMed Ag, Unomedical A / S, etc. The kit of the present invention can also contain one or more other components, including but not limited to instructions for use of the kit and / or its components, one or more other containers or compartments for containing the kit components, multiple containers or devices containing the preparation of the present invention, etc.

[0115] Examples

[0116] Some embodiments of the present disclosure will be described in more detail by way of specific examples. The following examples are provided for illustrative purposes only and are not intended to limit any aspect of the present invention in any way. For example, those skilled in the art will readily identify various non-critical parameters that can be changed or modified without undue experimentation to produce substantially the same results.

[0117] Example 1: Effect of Concentration on the Pharmacokinetics of Levothyroxine Liquid Formulations

[0118] The formulation work conducted by the present inventors resulted in stable levothyroxine liquid formulations that were prepared by dissolving sodium levothyroxine in acidified dimethyl sulfoxide (DMSO). These formulation studies are described in detail in U.S. Application No. 63 / 380090, entitled "Stable Levothyroxine Formulations in Aprotic Polar Solvents," filed on the same day as the present application, which is incorporated herein by reference in its entirety. Briefly, exemplary such formulations having the following composition were prepared: 10 mg / mL sodium levothyroxine; 5.53 weight / volume % trehalose dihydrate; 2.9 weight / volume % mannitol; 26 mM to 28 mM sulfuric acid (from a 1.0 N stock solution) and DMSO. When these solutions were prepared, it was found that the formulations did not freeze when stored under refrigerated conditions, and accelerated and real-time stability tests completed as of the filing date of the present application showed that the formulations were expected to achieve at least 2 years of refrigerated storage stability (using a 95% API content stability specification). When using 1.0 N H 2 SO 4 during preparation, the total water content of the formulation was approximately 5%. The water content of the formulation was almost entirely due to the addition of sulfuric acid (a minor contribution was from trehalose dihydrate), which had a water content of approximately 5 weight / weight % at 26 mM (from a 1.0 N (0.5 M) stock solution). When using concentrated acid (e.g., 10 N) during preparation, the total water content of the formulation could be reduced to approximately 1% without any significant effect on storage stability. Other exemplary formulations were prepared at a concentration of 5 mg / mL levothyroxine in DMSO acidified with approximately 13 mM to 15 mM sulfuric acid. These formulations were then used in preclinical studies of levothyroxine pharmacokinetics using experimental animals.

[0119] These studies were conducted in minipigs (male, 4 to 6 months old) obtained from Marshall Bioresources (Syracuse, NY). As previously described for other active pharmaceutical ingredients, XeriSol was used TMThe formulation platform (Xeris Pharmaceuticals, Inc.; Chicago, IL) prepared nine levothyroxine formulations, all of which contained 10 mg / mL of levothyroxine in acidified DMSO (see, for example, U.S. Patent Nos. 9,649,364, 10,485,850, and 11,020,403, the disclosures of which are incorporated herein by reference in their entireties). To study the effect of adding a sustained-release excipient to the formulation on pharmacokinetics, several formulations also received a certain concentration of poly(lactic-co-glycolic acid) (PLGA) with different molecular weights and concentrations, which is known to be useful for producing sustained-release formulations of various therapeutic compounds in XeriSol formulations (see U.S. Patent Publication No. 2021 / 0401945A1, which is incorporated herein by reference in its entirety). The prepared formulations are listed in Table 1 below.

[0120] Table 1 Test Formulations

[0121]

[0122] Miniature pigs (n = 12) were randomly divided into Treatment Group 1, Treatment Group 2, and Treatment Group 3 according to body weight, as shown in Table 2. Each of the three groups of four miniature pigs received a single dose of one of the levothyroxine formulations described in Table 2. The washout period was at least 21 days, followed by readministration of the animals with the next group of formulations. After the washout of each administration, the animals were transferred to the subsequent group as follows: Group 1 → Group 4 → Group 7; Group 2 → Group 5 → Group 8; Group 3 → Group 6 → Group 9. All formulations were administered subcutaneously at a fixed dose of 10 mg per animal. Blood samples were collected at the following time points for determination of plasma levothyroxine concentration: before administration (baseline), and 0.083 hours, 0.25 hours, 0.5 hours, 1 hour, 2 hours, 4 hours, 6 hours, 8 hours, 24 hours, 48 hours, 72 hours, 96 hours, 120 hours, 144 hours, 192 hours, 240 hours, 288 hours, 366 hours after administration.

[0123] Pyxant Labs Inc. developed a bioanalytical method for determining the concentration of levothyroxine (L-thyroxine) in miniature pig plasma. The method was qualified in the range of 2.00 ng / mL to 4000 ng / mL. The method used a surrogate matrix method for quantifying L-thyroxine, in which a 5% aqueous solution of bovine serum albumin (BSA) was used because L-thyroxine is endogenous. Samples were prepared by a protein precipitation extraction procedure. The extracts were analyzed by liquid chromatography / tandem mass spectrometry (LC-MS / MS) in positive ion mode under optimized conditions for detecting the positive ions of levothyroxine and thyroxine- 13 C6 (internal standard).

[0124] Non-compartmental pharmacokinetic analysis of the baseline-adjusted levothyroxine levels for each animal was performed using Phoenix 64 software version 8.2.0.4383 (Certara, Inc.; Princeton, NJ). The derived pharmacokinetic parameters included Cmax, Tmax, AUClast, AUC144-336, λz, t 1 / 2 and the values of AUC∞ were not determined due to high fluctuations in plasma levels during the terminal phase. The results of these analyses are presented in Figures 1 to 7.

[0125] All animals showed a quantifiable increase in the baseline-adjusted plasma levothyroxine concentration after SC administration of the test article (Figure 1). XeriSol TM The plasma concentration-time curve of levothyroxine (XS-1) was characterized by a gradual absorption with a median Cmax of 16 hours post-administration (range: 8 hours to 24 hours) (Figure 1 and Figure 3 ). Xerisol TM The PLGA levothyroxine formulations XS-3, formulation XS-8, and formulation XS-9 reached Cmax faster, with a median of 8 hours post-administration (range: 8 hours to 24 hours) (Figure 1 and Figure 3 ), while the PLGA levothyroxine formulations XS-2, formulation XS-4, formulation XS-5, formulation XS-6, and formulation XS-7 reached Cmax slower, with a median of 24 hours post-administration (range: 24 hours to 24 hours) (Figure 1 and Figure 3 ). The number of animals in each group was small (n = 4), and the Tmax ranges ( Figure 2 ) overlapped in all groups, and these differences were considered not significant.

[0126] As Figure 3 shown, Xerisol TM levothyroxine (XS-1) (1784.2 ± 527.1 ng / mL) and Xerisol TM levothyroxine PLGA formulations XS-2 (1750.1 ± 389.7) and formulation XS-9 (1789.3 ± 386.1 ng / mL) had the highest mean ± SD of baseline-adjusted levothyroxine Cmax. The Cmax of XS-8 was approximately 10% lower than that of XS-1, and the Cmax of the remaining formulations was 27% to 50% lower than that of XS-1, in the following order: XS-7 (1305.9 ± 305.9 ng / mL) > XS-4 (1156.9 ± 69.8 ng / mL) > XS-3 (1130.3 ± 504.2 ng / mL) > XS-5 (1011.2 ± 227.2 ng / mL) > XS-6 (896.6 ± 166.2 ng / mL).

[0127] As Figure 4 shown, the results indicate that the baseline-adjusted mean plasma levothyroxine exposure of Xerisol TM levothyroxine (XS-1) (96461.3 ± 17426.8) determined by AUClast (hour * ng / mL) was the highest, followed by XS-2 (83732.2 ± 36156.9). The AUClast of the five formulations was 20% to 32% lower than that of XS-1, in the following order: XS-7, XS-9, XS-5, XS-4, XS-8. The exposure levels of formulation XS-6 (56709.6 ± 3976.1) and formulation XS-3 (52934.8 ± 6383.5) were the lowest, 41% and 45% lower than that of formulation XS-1, respectively.

[0128] As Figure 5 shown, compared with XS-1, the mean ± SD of the partial exposure of baseline-adjusted plasma levothyroxine (AUC144-336) was higher in all PLGA-containing formulations. The AUC144-336 of formulation XS-2 (3655.1 ± 1903.9), formulation XS-3 (3075.3 ± 1182.2), and formulation XS-8 (2721.3 ± 3325.6) was 12% to 50% higher than that of formulation XS-1 (2435.8 ± 2710.5). The AUC144-336 of formulation XS-4 (4237.9 ± 1266.5), formulation XS-7 (4938.8 ± 2515.6), and formulation XS-9 (4472.6 ± 4503.2) was 74% to 103% higher than that of formulation XS-1. The partial exposure of formulation XS-5 (8467.0 ± 9782.1) and formulation XS-6 (7161.6 ± 4582.0) was 194% to 248% higher than that of formulation XS-1.

[0129] The following Table 2 further details these results, which compiles all measured and calculated pharmacokinetic parameters for each formulation.

[0130] Table 2 Pharmacokinetic parameters of baseline-adjusted plasma levothyroxine in minipigs after subcutaneous injection of XeriSol TM levothyroxine and XeriSol / PLGA levothyroxine formulations

[0131]

[0132] Formulations containing PLGA are expected to have a lower maximum concentration because, as PLGA degrades, drug release is expected to be slower. A significant decrease in Cmax was observed from formulation XS-3 to formulation XS-7. The expected Tmax may also be longer, but no substantial difference was observed between different formulations. The exposure of all formulations containing PLGA was 10% to 45% lower than that of XeriSol without PLGA TM levothyroxine (XS-1). These results suggest that the test articles containing PLGA are incompletely absorbed or absorbed very slowly. Tables 2 and Figure 5 The partial exposure AUC144-336 shown indicates that, compared with XS-1, the formulations containing PLGA have a greater partial exposure from 144 hours to 336 hours, suggesting that the formulations containing PLGA may have slow and extended absorption.

[0133] Next, the inventors wished to examine the pharmacokinetics of two Xerisol TM levothyroxine formulations containing 5 mg / mL sodium levothyroxine (“XP-8121-5”) and containing 10 mg / mL sodium levothyroxine (“XP-8121-10”). These formulations were prepared using the acidified DMSO solution method described above, in which an ionizing stabilizing excipient (sulfuric acid in this example) was added to the formulation in a molar ratio of approximately 2.5 to 1 relative to the API content in a DMSO-based solvent system. The formulations described in the previous section were administered to minipigs, and the pharmacokinetic parameters were determined as described above. The results are shown in Figures 6 and 7. In the unadjusted baseline (Figure 6) and adjusted baseline (Figure 7) analyses, both the 5 mg / mL formulation and the 10 mg / mL formulation had similar Tmax (within about 2 hours), whether evaluated on a linear graph ( Figure 6A and Figure 6B ) or a semi-logarithmic graph ( Figure 6B and Figure 7B ). Interestingly, in all analyses, the Cmax of the 5 mg / mL formulation was higher than that of the 10 mg / mL formulation, although the administration of the 10 mg / mL formulation resulted in a longer duration of levothyroxine plasma concentration compared with the 5 mg / mL formulation. These results suggest that there is a concentration effect on the release kinetics of levothyroxine from the injection site into the blood - levothyroxine administered from the 5 mg / mL solution is released into the bloodstream faster than that from the 10 mg / mL solution and reaches Cmax at an earlier time, but the latter shows a longer period of elevated levothyroxine blood level after injection compared with the former.

[0134] The unadjusted baseline or adjusted baseline individual pharmacokinetic parameters and overall pharmacokinetic parameters determined as described above are shown in Tables 3 and 4, respectively.

[0135]

[0136]

[0137] In summary, these results indicate that there is a dose - dependent or concentration - dependent effect for the release of levothyroxine from the subcutaneous injection site into the bloodstream. Specifically, at a given injection volume, a higher concentration of levothyroxine in the formulation for injection provides a means to reach the therapeutic level of levothyroxine in plasma, which subsequently has a longer - lasting concentration compared to a lower - concentration formulation (e.g., a 10 mg / mL levothyroxine formulation shows a longer - lasting elevated plasma levothyroxine level compared to a 5 mg / mL formulation). Based on these results, the more persistent delivery of levothyroxine over time using such formulations should be able to reduce the administration frequency of levothyroxine in subjects being treated for hypothyroidism with this drug, from daily administration (such as currently with oral solid dosage forms containing levothyroxine, e.g., ) to a lower - frequency subcutaneous administration (e.g., using higher concentrations of levothyroxine, such as 15 mg / mL, 20 mg / mL, 40 mg / mL, etc., up to the solubility limit of the API in a non - protonic polar solvent system, once a week, or possibly once every other week or even less frequently).

[0138] Example 2: Concentration Effect on the Pharmacokinetics of a Diazepam - containing Liquid Formulation

[0139] To further examine the concentration effect on pharmacokinetics observed in Example 1, another compound, diazepam, which is poorly soluble in an aqueous medium, was formulated in the XeriSol TM acidified DMSO solution as described in Example 1 for levothyroxine, at concentrations of 50 mg / mL and 100 mg / mL. Then, as described for levothyroxine in Example 1 above, the pharmacokinetics of these formulations were evaluated in minipigs after SC administration (Xeris Pharmaceuticals, Inc. Study 12412). The results of these studies are as Figure 8 shown.

[0140] The pharmacokinetic characteristics of the higher (100 mg / mL) diazepam concentration formulation were lower mean Cmax (SD) (296 ng / mL (182 ng / mL) vs. 412 ng / mL (454 ng / mL)) and longer median (range) Tmax (240 minutes (60 minutes to 240 minutes) vs. 120 minutes (15 minutes to 20 minutes)) compared to the lower (50 mg / mL) diazepam concentration formulation. The mean (SD) diazepam exposure (AUClast) was similar for the 100 mg / mL formulation (102752 ng*min / mL (39313 ng*min / mL)) and the 50 mg / mL formulation (99775 ng*min / mL (54804 ng*min / mL)).

[0141] Thus, as shown by the observations for levothyroxine in Example 1, the diazepam-containing liquid injection formulation showed a dose-dependent or concentration-dependent effect on the release of diazepam from the subcutaneous injection site into the blood. Specifically, at a given injection volume, a higher concentration of diazepam in the formulation for injection provided a means to rapidly achieve the therapeutic level of diazepam in plasma, with subsequent sustained concentration and slower clearance kinetics compared to the lower concentration formulation (e.g., the 100 mg / mL diazepam formulation showed more prolonged sustained release kinetics compared to the 50 mg / mL formulation). Using such a diazepam formulation that delivers continuously over time should allow for a reduction in the frequency of diazepam administration in subjects being treated with the drug, from daily oral administration to a lower frequency (e.g., weekly) subcutaneous administration, for example, applicable to diseases or physical conditions treated with benzodiazepines such as diazepam, such as anxiety disorders, muscle spasms, and seizures.

[0142] Example 3: Concentration Effect on the Pharmacokinetics of a Tamsulosin-Containing Liquid Formulation

[0143] Another active pharmaceutical ingredient with solubility characteristics similar to levothyroxine and diazepam is tamsulosin, a drug currently administered orally once daily for the treatment of certain urogenital diseases / conditions in men, particularly benign prostatic hyperplasia and kidney stones. Tamsulosin has good solubility in DMSO (100 mM, >40 mg / ml) and poor solubility in water / physiological environment (about 10 mM, 4 mg / ml). The drug is typically administered to patients at a dose of 0.4 mg per day, which is equivalent to approximately 3 mg per week. Thus, as described for levothyroxine and diazepam in Example 1 and Example 2, tamsulosin was formulated in an acidified DMSO solution of XeriSol TM Then, a 0.1 ml volume of 30 mg / mL XeriSol containing tamsulosin was subcutaneously injected into patients TMTamsulosin formulations are thus administered to a patient in an amount of 3 mg per injection. Such an amount would far exceed the solubility limit of tamsulosin in a subcutaneous aqueous environment, and tamsulosin would precipitate substantially, thereby creating a depot for sustained delivery of tamsulosin, the pharmacokinetics of which would be similar to those of levothyroxine and diazepam described above. Thus, after using such formulations with a more extended sustained release time of tamsulosin, it should be possible to reduce the frequency of tamsulosin administration in subjects being treated with tamsulosin, from daily oral administration to a lower frequency (e.g., weekly) of subcutaneous administration.

[0144] Example 4: Human Pharmacokinetic Study Using a Liquid Formulation Containing Levothyroxine

[0145] In another set of studies, the inventors of the present invention wished to determine whether the pharmacokinetics observed in humans using the XP-8121 levothyroxine-containing formulation of the present invention were similar to those observed in preclinical studies using experimental animals as described in Example 1. These studies were designed for several purposes: (a) to characterize the pharmacokinetics of subcutaneous injection of XP-8121 (doses of 600 μg, 1200 μg, and 1500 μg, XP-8121 from the 10 mg / mL levothyroxine formulation described in Example 2); (b) to evaluate the bioavailability of XP-8121 (relative to an oral levothyroxine formulation, such as ) and the dose proportionality (600 μg, 1200 μg, and 1500 μg); and (c) to evaluate the safety and tolerability of XP-8121. A subsequent study of PK analysis (population PK model) was also conducted to compare the steady-state exposures (AUC) of weekly subcutaneous (SC) administration of XP-8121 and daily oral (PO) administration and to examine the dose conversion from PO to XP-8121 SC.

[0146] This study was designed according to the schematic diagram shown Figure 9 This study, designated XP-8121-108, examined the plasma levothyroxine pharmacokinetics in 30 subjects, who were evenly divided into two groups of 15 each, and received 600 μg of PO or XP - 8121SC, followed by a 42 - day washout period, after which the subjects received 600 μg of levothyroxine in another form, and then after another 42 - day washout period, both groups received a subcutaneous injection of 1200 μg of XP - 8121. After a brief pause in the study and review of safety and pharmacokinetic data by the US FDA, the study protocol was amended to evaluate levothyroxine at doses above 1200 μg. Thus, in the final part of the study, an additional 30 subjects received a subcutaneous injection of 1500 μg of levothyroxine in the XP - 8121 formulation of the present invention. Pharmacokinetic parameters were measured and determined as shown in Figure 1 and calculated as described in the pre - clinical pharmacokinetic analysis of Example 1. The results of these studies are shown in Figures 10 to 14.

[0147] Figure 10 shows the mean baseline - adjusted plasma levothyroxine concentration over time following subcutaneous (SC) administration of different doses (600 μg, 1200 μg, 1500 μg) of XP - 8121 and oral (PO) administration of 600 μg of compared to, plotted as a line graph ( Figure 10A ) or a log graph ( Figure 10B ). As Figure 10A and Figure 10B shown, PO 600 μg shows a rapid increase followed by a rapid decline in levothyroxine levels, while XP - 8121SC (all doses) shows a slow increase in levothyroxine levels followed by a sustained exposure above the levothyroxine levels obtained with PO. In addition, after reaching Cmax, the plasma T4 levels of XP - 8121 remain at a stable high level for 4 to 5 days, in contrast, they decline immediately. Finally, the Tmax of XP - 8121 also appears to be longer than that of PO.

[0148] Similar results were obtained for other pharmacokinetic parameters. As Figure 11A shown, the median Tmax of XP - 8121 (48 hours, 72 hours, 96 hours for doses of 600 μg, 1200 μg, and 1500 μg respectively) is greater than the observed median Tmax of PO (3 hours). The mean half - life (T 1 / 2 ) of XP - 8121 (177 hours, 112 hours, and 120 hours for doses of 600 μg, 1200 μg, and 1500 μg respectively) is similar to the observed mean half - life (T 1 / 2 ) of PO (144 hours) ( Figure 11B ). Figure 12 shows the use of SC XP - 8121 compared to PO The mean baseline levothyroxine exposure in the subjects being treated. The mean Cmax of XP-8121 600 μg was lower than the mean Cmax of PO 600 μg ( Figure 12A ). In addition, although the unadjusted mean levothyroxine AUClast of XP-8121 SC 600 μg and PO 600 μg were similar (data not shown), the baseline-adjusted mean levothyroxine AUClast of XP-8121 SC 600 μg was 35% higher than that of PO 600 μg ( Figure 12B ). XP-8121 SC also showed a dose-dependent increase in exposure (Cmax ( Figure 12A ) and AUClast ( Figure 12B ). Thus, compared with PO, single-dose SC XP-8121 had a significantly lower Cmax and similar or higher exposure as determined by AUClast.

[0149] To further examine the dose proportionality of baseline-adjusted XP-8121, the baseline-adjusted mean Cmax was normalized according to the levothyroxine dose and plotted on a linear scale ( Figure 13A ) or a logarithmic scale ( Figure 13B ). These parameters were calculated from the concentrations at the PK time points matched in Study Parts 1 and 2 (PK population), as Figure 9 shown. The results shown in Figure 13 indicate that the baseline-adjusted Cmax of XP-8121 is proportional to the dose. Similarly, it was also found that the baseline-adjusted AUClast of XP-8121 is proportional to the dose, whether plotted on a linear scale ( Figure 14A ) or a logarithmic scale ( Figure 14B ).

[0150] Combining those foregoing examples, these results indicate that storage-stable, ready-to-use injectable levothyroxine liquid formulations can be produced as described herein, and that these formulations, when administered subcutaneously to a subject (such as the human in Example 4), can promote an increase in the plasma levothyroxine level of the subject, with slower release kinetics (i.e., extended-release pharmacokinetics) compared to oral levothyroxine immediate-release solid formulations such as observed. More specifically, the results of Example 3 indicate that PO 600 μg exhibited a rapid rise and subsequent rapid decline in levothyroxine level, while XP-8121 SC (all doses) exhibited a slow rise and subsequent sustained exposure in levothyroxine level. In addition, the Tmax of XP-8121 SC (all doses) was longer than PO 600 μg. The plasma half-life of levothyroxine was similar in all groups, while the mean Cmax of XP-8121 600 μg was less than half of the Cmax of PO 600 μg (47%). In addition, the increase in levothyroxine exposure (Cmax and AUClast) of XP-8121 SC was considered to be proportional to the dose. Thus, as was seen in the animal studies of Example 1 and Example 2 above, there is a dose-dependent or concentration-dependent effect of the release of levothyroxine from the subcutaneous injection site into the human bloodstream. Specifically, subcutaneous injection of a higher dose of levothyroxine in humans can bring plasma levothyroxine to a therapeutic level, and compared with injection of a lower dose of levothyroxine, the subsequent concentration is sustained and the clearance kinetics are slower (e.g., injection of 1500 μg of levothyroxine showed longer sustained-release kinetics compared with injection of 1200 μg of levothyroxine; they showed longer sustained-release kinetics compared with a levothyroxine preparation at a dose of 600 μg, and these in turn showed longer sustained-release kinetics compared with oral levothyroxine at a dose of 600 μg ). Based on these results, more persistent delivery of levothyroxine over time using such a formulation should be able to reduce the administration frequency of levothyroxine in subjects being treated for hypothyroidism with this drug, from daily administration (such as current oral solid dosage forms containing levothyroxine, e.g., ) to a lower frequency of subcutaneous use (e.g., once a week).

[0151] All embodiments of the present invention, including the compositions and / or methods disclosed and claimed herein, can be made and implemented in accordance with this disclosure without undue experimentation. Although the compositions and methods of this disclosure have been described in accordance with certain exemplary embodiments, it will be apparent to those skilled in the art that changes can be made to the compositions and methods and to the steps or the order of the steps of the methods described herein without departing from the concept, spirit, and scope of the invention. More specifically, it is apparent that specific reagents, both chemically and physiologically relevant, can be substituted for the reagents described herein while achieving the same or similar results. All such similar substitutions and alterations that are apparent to those skilled in the art are considered to be within the spirit, scope, and concept of the invention as defined by the appended claims.

Claims

1. A storage-stable sustained-release therapeutic preparation, comprising: (a) at least one therapeutic agent; (b) at least one ionization-stabilizing excipient; and (c) an aprotic polar solvent system, wherein the at least one therapeutic agent is poorly soluble in an aqueous environment but readily soluble in the aprotic polar solvent system, wherein the preparation is stably stored at 2°C to 8°C for at least six months, and wherein when administered to a patient, as compared to an immediate-release preparation comprising the at least one therapeutic agent, the storage-stable sustained-release therapeutic preparation results in an extended period of presence of at least one therapeutic agent at a therapeutic level in the blood of the patient.

2. The preparation according to claim 1, wherein the therapeutic agent is selected from levothyroxine or its salts, benzodiazepines and tamsulosin.

3. The preparation according to claim 2, wherein the therapeutic agent is levothyroxine free acid or levothyroxine sodium.

4. The preparation according to claim 2, wherein the benzodiazepine drug is diazepam.

5. The preparation according to claim 1, wherein the aprotic polar solvent system comprises or is dimethyl sulfoxide (DMSO).

6. The preparation according to claim 1, wherein the ionization-stabilizing excipient is a mineral acid.

7. The preparation according to claim 6, wherein the mineral acid is selected from hydrochloric acid, sulfuric acid, nitric acid, and phosphoric acid.

8. The preparation according to claim 7, wherein the mineral acid is sulfuric acid.

9. The preparation according to claim 6, wherein the mineral acid is present in the storage-stable sustained-release therapeutic preparation at a molar concentration ratio of 2:1 to 3:1 relative to the concentration of the therapeutic agent in the storage-stable sustained-release therapeutic preparation.

10. The preparation according to claim 9, wherein the mineral acid is present in the storage-stable sustained-release therapeutic preparation at a molar concentration ratio of about 2.5:1 relative to the concentration of the therapeutic agent in the storage-stable sustained-release therapeutic preparation.

11. The preparation according to claim 3, wherein the storage-stable sustained-release therapeutic preparation comprises levothyroxine sodium at a concentration of about 5 mg / mL and sulfuric acid at a concentration of about 13 mM to about 15 mM.

12. The preparation according to claim 3, wherein the storage-stable sustained-release therapeutic preparation comprises levothyroxine sodium at a concentration of about 10 mg / mL and sulfuric acid at a concentration of about 26 mM to about 28 mM.

13. The preparation according to claim 1, which further comprises one or more other pharmaceutically acceptable excipients.

14. The preparation according to claim 13, wherein the one or more other pharmaceutically acceptable excipients are selected from sugars, sugar alcohols, preservatives, and polymers.

15. The preparation according to claim 14, wherein the sugar is trehalose or a salt or hydrate thereof.

16. The preparation according to claim 15, wherein the sugar is trehalose dihydrate.

17. The preparation according to claim 14, wherein the sugar alcohol is mannitol.

18. The preparation according to claim 14, wherein the preservative is benzyl alcohol.

19. The preparation according to claim 14, wherein the polymer is poly(lactic-co-glycolic acid) (PLGA).

20. The preparation according to claim 19, wherein the PLGA is ester-terminated PLGA or acid-terminated PLGA.

21. A method of treating or preventing a disease or disorder in a human or veterinary animal, the method comprising introducing a therapeutically effective amount of the preparation according to claim 1 into a human or veterinary animal in need thereof, monitoring the level of the preparation in the plasma of the human or veterinary animal over time, and repeating the introduction of the preparation into the subject as needed to treat or prevent the disease or disorder in the human or veterinary animal.

22. The method according to claim 21, wherein the preparation is introduced into the human or veterinary animal by parenteral administration.

23. The method according to claim 21, wherein the parenteral administration is by injection or infusion.

24. The method according to claim 23, wherein the injection is subcutaneous, intradermal or intramuscular injection.

25. The method according to claim 23, wherein the injection is subcutaneous injection.

26. The method according to claim 23, wherein the infusion is accomplished by pump infusion.

27. The method according to claim 26, wherein the pump infusion comprises continuous pump infusion or rapid pump infusion or a combination thereof.

28. The method according to claim 21, wherein the therapeutic agent is selected from levothyroxine or its salts, benzodiazepines and tamsulosin.

29. The method according to claim 28, wherein the therapeutic agent is levothyroxine free acid or levothyroxine sodium.

30. The method according to claim 29, wherein the disease or disorder is hypothyroidism or a disease or disorder associated with or characterized by hypothyroidism.

31. The method according to claim 30, wherein the disease or disorder associated with or characterized by hypothyroidism is thyroiditis, Hashimoto's disease, myxedema or myxedema coma / crisis.

32. The method according to claim 28, wherein the therapeutic agent is a benzodiazepine drug.

33. The method according to claim 32, wherein the benzodiazepine drug is diazepam.

34. The method according to claim 32, wherein the disease or disorder is anxiety disorder, muscle spasm or seizure.

35. The method according to claim 28, wherein the therapeutic agent is tamsulosin.

36. The method according to claim 35, wherein the disease or disorder is benign prostatic hyperplasia or kidney stone.

37. A method of preparing a storage-stable sustained-release therapeutic preparation of at least one therapeutic agent, the method comprising mixing at least one ionization-stable excipient, an aprotic polar solvent system and at least one therapeutic agent to form a storage-stable sustained-release therapeutic preparation, the at least one therapeutic agent being poorly soluble in an aqueous environment and readily soluble in the aprotic polar solvent system, such that when administered to a human or veterinary animal, the storage-stable sustained-release therapeutic preparation results in an extended period of presence of at least one therapeutic agent at a therapeutic level in the blood of the human or veterinary animal, as compared to an immediate-release preparation comprising the at least one therapeutic agent.

38. The method according to claim 37, wherein the therapeutic agent is selected from levothyroxine or its salts, benzodiazepines and tamsulosin.

39. The method according to claim 38, wherein the therapeutic agent is levothyroxine free acid or levothyroxine sodium.

40. The method according to claim 38, wherein the benzodiazepine drug is diazepam.

41. The method according to claim 37, wherein the aprotic polar solvent system comprises or is dimethyl sulfoxide (DMSO).

42. The method according to claim 37, wherein the ionization-stable excipient is a mineral acid.

43. The method according to claim 42, wherein the mineral acid is selected from hydrochloric acid, sulfuric acid, nitric acid and phosphoric acid.

44. The method according to claim 43, wherein the mineral acid is sulfuric acid.

45. The method according to claim 42, wherein the mineral acid is present in the storage-stable sustained-release therapeutic formulation in a molar concentration ratio of 2:1 to 3:1 relative to the concentration of the therapeutic agent in the storage-stable sustained-release therapeutic formulation.

46. The method according to claim 45, wherein the mineral acid is present in the storage-stable sustained-release therapeutic formulation in a molar concentration ratio of about 2.5:1 relative to the concentration of the therapeutic agent in the storage-stable sustained-release therapeutic formulation.

47. The method according to claim 39, wherein the storage-stable sustained-release therapeutic formulation comprises levothyroxine sodium at a concentration of about 5 mg / mL and sulfuric acid at a concentration of about 13 mM to about 15 mM.

48. The method according to claim 39, wherein the storage-stable sustained-release therapeutic formulation comprises levothyroxine sodium at a concentration of about 10 mg / mL and sulfuric acid at a concentration of about 26 mM to about 28 mM.

49. The method according to claim 37, further comprising one or more other pharmaceutically acceptable excipients.

50. The method according to claim 49, wherein the one or more other pharmaceutically acceptable excipients are selected from sugars, sugar alcohols, preservatives, and polymers.

51. The method according to claim 50, wherein the sugar is trehalose or its salt or hydrate.

52. The method according to claim 51, wherein the sugar is trehalose dihydrate.

53. The method according to claim 50, wherein the sugar alcohol is mannitol.

54. The method according to claim 50, wherein the preservative is benzyl alcohol.

55. The method according to claim 50, wherein the polymer is poly(lactic-co-glycolic acid) (PLGA).

56. The method according to claim 55, wherein the PLGA is ester-terminated PLGA or acid-terminated PLGA.

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