Injectable high concentration pharmaceutical formulations and methods of making and using same

Abstract

The present invention provides compositions comprising one or more active pharmaceutical ingredients in the form of a paste with a high solids content that can be injected into animals in relatively small amounts using standard commercially available syringes. The present invention also provides methods for making such compositions, particularly compositions comprising high molecular weight active ingredients (e.g., antibodies, enzymes, and other proteins and peptides) at relatively high therapeutic concentrations in a paste with a high solids content. The present invention further provides methods for using such formulations in treating, preventing, and / or ameliorating certain diseases and disorders in animals, including humans in need thereof. The present invention also provides kits comprising the formulations of the present invention and a suitable syringe, which in some aspects may be preloaded or prefilled with the composition of the present invention.

Description

[Technical field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application claims the benefit of priority to U.S. Provisional Patent Application No. 63 / 242,405, filed September 9, 2021, and U.S. Provisional Patent Application No. 63 / 351,786, filed June 13, 2022, the disclosures of which are incorporated by reference herein in their entireties.

[0002] Statement of Prior Disclosure by Inventor or Co-Inventors Some of the materials disclosed herein are disclosed in U.S. Patent Nos. 8,110,209, 8,790,679 and 9,314,424, and U.S. Patent Application Publication No. 2017 / 0216529.

[0003] FIELD OF THEINVENTION The present invention generally relates to parenteral, i.e., intradermal, subcutaneous and / or intramuscular injection of pharmaceutical formulations containing at least one active pharmaceutical ingredient in high concentrations, particularly in the form of a paste, and provides such formulations, methods of making and using such formulations, and kits containing such formulations. [Background technology]

[0004] 2. Description of Related Art Parenteral injection refers to the administration of a drug, agent or vaccine via injection under or through one or more layers of the skin or mucosa of an animal. Standard injections are given to the subcutaneous or intramuscular regions of an animal, e.g., a human patient. These deeper sites are targeted because the tissue expands more readily than shallow skin sites to accommodate the 0.1-3.0 cc (ml) injection volumes required to deliver most therapeutic agents.

[0005] In general, injections have been classified into various categories, including: (1) ready-to-inject solutions; (2) dry soluble products (solutes) that can be combined with a solvent immediately prior to injection into a patient; (3) dry insoluble products that can be combined with a suitable injection vehicle prior to administration; (4) ready-to-inject suspensions; and (5) ready-to-inject emulsions. Such injectable formulations are administered by routes including intravenous, subcutaneous, intradermal, intramuscular, intraspinal, intracisternal, and intrathecal. The nature of the therapeutic agent and the disease or disorder being treated will readily dictate the route of administration. However, the desired route of administration constrains the therapeutic formulation itself. For example, solutions intended for subcutaneous administration require careful attention to isotonicity to avoid irritation to nerves and tissues in the area surrounding the injection. Similarly, suspensions are not administered directly into the bloodstream due to the possibility that insoluble particles may block capillaries.

[0006] Compared to other dosage forms and routes of administration (e.g., oral, transdermal), injectables have certain advantages, including immediate physiological action (e.g., via intravenous injection), avoidance of intestinal absorption problems associated with many drugs, and precise administration of the desired dose into the patient's bloodstream. On the other hand, one of the disadvantages of injectables is the pain and discomfort present at the administration site associated with certain pharmacoactive agents, and the trauma of needle insertion subcutaneously or intravenously. Each injection administered causes some degree of discomfort to the patient.

[0007] Currently, biopharmaceuticals are typically reconstituted into a sterile solution and administered into the subcutaneous or intramuscular region using a large gauge needle, for example, in the range of 18-30 gauge. Pain is caused by, among other things, the depth of needle penetration, the size of the "gauge" of the needle, the large volume injected, and diffusion of the drug from the injection site. In addition to the issues with pain associated with administering injectables, current embodiments of injections also have other drawbacks. For example, many proteins and sustained release drugs must be reconstituted immediately prior to administration. Drug administration can be inflexible and imprecise. Furthermore, many formulations must be refrigerated to protect the drug from physical and / or chemical degradation (e.g., hydrolysis). Additionally, current administration systems are wasteful in that the injection device retains a significant amount of drug product. Furthermore, injectable formulations must typically be concentrated and stabilized to deliver the required amount required. Standard injectables are given in liquid form. Products sold as liquids or lyophilized powders must be reconstituted with an aqueous carrier prior to injection. Many therapeutic protein and vaccine products are manufactured in dry solid form to enhance stability during storage. Prior to injection, these formulations are diluted / reconstituted in a pharma- ceutically acceptable vehicle, including sterile water for injection (SWFI), phosphate buffer, or isotonic saline, until a solution or suspension is obtained.

[0008] More recently, the preparation and use of highly concentrated pharmaceutical formulations in the form of low water suspensions, colloids or pastes has been described (see, e.g., U.S. Pat. Nos. 8,110,209, 8,790,679 and 9,314,424, all of the disclosures of which are incorporated herein by reference in their entireties, and U.S. Patent Application Publication No. 2017 / 0216529). Such formulations contain the active pharmaceutical ingredient at substantially higher concentrations than found in conventional aqueous pharmaceutical formulations. Pastes, i.e., biphasic mixtures of solids dispersed in a non-solvent liquid, can be effective dosage forms (e.g., compared to solids) for drug delivery (e.g., intradermally). For example, pastes can potentially achieve much higher solids (e.g., drug) concentrations than typical solutions (e.g., aqueous solutions), while also providing superior stability compared to aqueous solutions because the active ingredient in the paste can be formulated in the solid state (e.g., as a powder). This approach can be particularly advantageous for formulating active pharmaceutical ingredients that are poorly soluble in aqueous solutions or that are prone to chemical degradation (e.g., hydrolysis) and / or physical instability (e.g., aggregation) when formulated into low-concentration, highly aqueous formulations for delivery to patients.

[0009] A paste is a semi-solid dosage form that contains a high percentage of finely dispersed solids (e.g., powder particles) in an oily substance (e.g., oil or hydrocarbon base) with a relatively hard and high consistency. The actual solids content (or solids concentration - in both cases describing the amount of solids in the formulation, "solids content" refers to the weight percent of solids relative to the total weight of the formulation (solids plus liquids), while "solids concentration" refers to the concentration of solids per unit volume in the formulation (e.g., g / mL, mg / mL, etc.)) of a paste mainly depends on the characteristics of the constituent powders and may be less than or greater than those in the USP-NF definition (see, for example, U.S. Patent No. 8,110,209 (Patent Document 1), U.S. Patent No. 8,790,679 (Patent Document 2), U.S. Patent No. 9,314,424 (Patent Document 3), and U.S. Patent No. 11,129,940 (Patent Document 5), all of the disclosures of which are incorporated herein by reference in their entirety). To prepare a paste, the minimum amount of fluid added to the powder must be sufficient to coat the powder particles. In an ideal situation, all powder-powder contacts are completely blocked and each powder particle is not glued / attached / agglomerated with any other particle. However, in practice, many finely divided powders are highly cohesive and even when high shear mixing techniques are applied, it may not be possible to completely block all direct powder-powder contacts. Additional fluid is then added to the mixture to fill the gaps (i.e., void volume) between the powder particles, allowing the particles to flow as a fluid when the yield stress of the paste is exceeded. Thus, powders with very low density (i.e., powders with a high surface area to volume ratio) require more fluid to form a paste than powders with a lower surface area to volume ratio. Thus, as discussed further, the percent solids content of a paste may vary widely and may vary depending on multiple factors, including the process by which the paste is prepared (e.g., non-limiting examples include freeze drying, spray drying, spray freeze drying, thin film freezing, solvent extraction / exchange, coacervation, and further particle engineering techniques known in the art).

[0010] Although often classified as a suspension because they are two-phase systems (containing both a solid (particulate) phase dispersed in a liquid (diluent / non-solvent) phase), pastes are physically different from traditional suspensions, and other formulations with high solids concentrations, such as gels, in that the concentration of particulate matter (e.g., powder) in the composition is high enough to prevent the particles from settling in the fluid over the storage conditions and periods associated with commercial pharmaceuticals. This gives pastes a firm consistency, making them highly viscous, compared to gels, creams, foams, and other "semi-solid" pharmaceutical dosage forms.

[0011] Thus, parenteral (e.g., intradermal, subcutaneous and / or intramuscular) delivery (e.g., injection) of such pastes can be difficult. In particular, such pastes typically have a significantly higher apparent viscosity when compared to conventional aqueous solutions, and injection of such highly viscous pastes using conventional syringes is generally considered difficult, if not impossible (e.g., requiring excessive force and / or causing excessive pain, e.g., due to the use of large needles). Furthermore, being two-phase mixtures of liquids containing homogeneously dispersed particulate matter, these compositions are particularly susceptible to partial and / or complete clogging of the delivery device, resulting in further limitations to the possibility of intradermal delivery of therapeutic pastes.

[0012] Methods for injecting pastes have been disclosed previously. For example, US Patent No. 8,790,679, US Patent No. 8,110,209 and US Patent No. 9,314,424, as well as US Patent Application Publication No. 2017 / 0007675 and US Patent Application Publication No. 2017 / 0216529 (all of which disclosures are incorporated herein by reference in their entirety), disclose the preparation of therapeutic pastes for intradermal administration, showing that paste formulations typically have poor flow characteristics in standard syringes, and therefore require novel needle / syringe designs to deliver such formulations. To achieve delivery, the injection device preferably incorporates a plunger that can fit into the needle lumen and acts such that the entire amount of pharmaceutical formulation loaded into the device is loaded into the needle lumen and pushed out to the patient during administration using a positive displacement design. However, this particular type of configuration would require a plunger that fits within the lumen of the needle and that, upon actuation, moves toward the end of the needle in a manner such that substantially all (e.g., approaching or equal to 100%) of the loaded pharmaceutical formulation is expelled through the needle to the injection site.

[0013] As is well known in the art, commercially available syringes have an inner diameter several times larger than the inner diameter of the needle lumen. For example, a standard 1 mL long syringe used in many commercially available injectable pharmaceutical products has an inner diameter of approximately 6.4 mm (compared to approximately 0.26 mm for a 25 G needle). Furthermore, the injection devices described in the prior art are only capable of delivering very small amounts of paste and / or fluid through a standard needle. As an example, a typical needle used for subcutaneous injection is a 27 gauge (i.e., 27 G) ultra-thin wall (UTW) 6 mm long needle. This needle has an inner diameter of approximately 300 μm (0.300 mm). If the internal volume of the needle is modeled as a cylinder with a height of 6 mm and a diameter of 0.300 mm, the amount of paste that can be contained inside such a needle is 4.24×10 -4 cm 3, or approximately 0.42 μL. Typical injection volumes for intradermal delivery are often in the range of 100-1000 μL (0.1-1.0 mL), with larger volumes to be delivered (e.g., 2000 or 3000 μL), depending on the indication, drug, etc. Thus, a very long and very large (in terms of internal diameter) needle would be required to deliver the most therapeutically relevant volumes.

[0014] As further discussed in the art, "the needle portion of the injection device is about 6 to about 8 cm in length, thereby providing a lumen with an internal volume sufficient to accommodate a dose of the semi-solid therapeutic formulation and a plunger." U.S. Patent Application Publication No. 2006 / 0211982 (Patent Document 7), paragraph

[0115] . Typical needle lengths for intradermal (ID) and subcutaneous (SC) administration are ≥ 0.5 inches (i.e., 1.3 cm). For deeper intramuscular (IM) injections, needles of only 1.0 to 1.5 inches (i.e., 2.5 to 3.8 cm) are commonly employed. Thus, needles envisioned for administration of viscous therapeutic pastes would have to be at least twice as long as commercially available needles. However, even with these long and specially designed needles, and assuming a relatively large internal diameter, the amount that can be placed within the lumen may still be significantly below the amount required to achieve a therapeutic dose. For example, an 8 cm long 18G needle (0.84 mm internal diameter) has an internal volume of only 4.4 × 10 -2 cm 3 , or approximately 44 μL.

[0015] In addition to the small amounts that can be administered from configurations in which the entire dose is contained within the needle lumen, such long needles typically must be specially manufactured and may be frightening or repulsive to certain patients due to their length. Furthermore, because the pain of an injection may be related to the overall diameter (gauge) of the needle, such large needles can be very painful, thus adversely affecting patient compliance with administration regimens that require multiple injections with such large needles.

[0016] Thus, there is a need in the art for storage-stable compositions, methods, kits and devices for use in parenteral delivery of highly concentrated, viscous non-Newtonian fluids (such as pastes) containing high concentrations of one or more therapeutic agents, particularly therapeutic agents that are themselves relatively high molecular weight (e.g., biologics including antibodies (monoclonal and / or polyclonal) and fragments or complexes thereof, vaccines, enzymes, receptor agonistic or antagonistic peptides and proteins, oligonucleotides and vectors containing them, etc.), using a standard syringe connected to a needle typically used for administration. There is a further need for compositions, methods, kits and / or devices for the delivery of such therapeutic fluids (including pastes) in amounts that may exceed the volume of the needle lumen. [Prior art documents] [Patent documents]

[0017] [Patent Document 1] U.S. Patent No. 8,110,209 [Patent Document 2] U.S. Patent No. 8,790,679 [Patent Document 3] U.S. Patent No. 9,314,424 [Patent Document 4] US Patent Application Publication No. 2017 / 0216529 [Patent Document 5] U.S. Patent No. 11,129,940 [Patent Document 6] US Patent Application Publication No. 2017 / 0007675 [Patent Document 7] US Patent Application Publication No. 2006 / 0211982 Summary of the Invention

[0018] The present invention provides compositions suitable for parenteral administration, i.e., intradermal, subcutaneous and / or intramuscular administration, of highly concentrated pharmaceutical formulations, and provides such formulations, methods of preparation and use of such formulations, and kits containing such formulations. Certain aspects of the invention described herein are directed to the discovery that non-Newtonian fluids and viscoelastic semi-solid compositions, such as pastes (and even high viscosity Newtonian fluids), containing high concentrations of active pharmaceutical ingredients can be easily delivered parenterally from standard (i.e., commercially available) syringe / needle combinations. Thus, the present invention provides pharmaceutical formulations that contain a large amount of active pharmaceutical ingredient in a relatively small amount of diluent or carrier (compared to conventional aqueous pharmaceutical formulations), and in particular, pharmaceutical formulations that are manufactured in a manner that allows the formulation to be administered to a patient other than intravenously, e.g., subcutaneously, intradermally or parenterally, that provides a ready-to-use formulation (i.e., one that does not need to be reconstituted or diluted before administration to a patient), and that can also provide longer shelf life than previously achieved. Thus, the present invention facilitates the manufacture, storage, and parenteral delivery of drugs that were previously only delivered intravenously - i.e., instead of large volumes of drug (regardless of route of administration), smaller intradermal, subcutaneous, or intramuscular injections can be used to achieve the same therapeutic effect. In certain embodiments, this approach is coupled with a reduction in injection site adverse reactions that are often associated with parenteral injection of large volumes of pharmaceutical formulations.

[0019] In one aspect, the invention provides a method for producing a high concentration / high viscosity injectable formulation for parenteral injection of a therapeutic agent or active pharmaceutical ingredient in the form of a high solids content paste (e.g., a formulation having an apparent viscosity of more than about 50 cP, more than about 100 cP, more than about 200 cP, or more than about 250 cP). A particular method according to this aspect of the invention includes spray drying and freeze drying an aqueous formulation containing one or more active pharmaceutical ingredients, and then processing (e.g., milling, sieving, etc.) the resulting powder to break up larger aggregates and produce powders and powder particles with relatively small diameters and narrow size distributions so that they can be delivered through small diameter needles suitable for administration by parenteral injection. Such powders are then mixed with one or more non-solvent diluents to produce a high solids content and high active ingredient concentration paste formulation suitable for injection in small volumes into an animal (e.g., a human or veterinary animal) to treat, ameliorate, prevent, or diagnose a disease or disorder in the animal. The invention also provides such paste formulations produced by such methods of the invention.

[0020] In a further aspect, the present invention provides highly concentrated / viscosity injectable formulations for parenteral (e.g., intradermal, subcutaneous and / or intramuscular) administration of therapeutic agents or active pharmaceutical ingredients, as well as methods for producing such formulations in a manner that results in highly concentrated / viscosity, storage stable, ready-to-use (i.e., no need for reconstitution and / or dilution prior to use) formulations. For purposes of the present invention, a "therapeutic agent" or "active pharmaceutical ingredient" or "pharmaceutical active ingredient" (these terms are used interchangeably and equivalently herein and will be readily understood by those of skill in the art) includes drugs, vaccines, hormones (particularly peptide hormones, e.g., insulin, glucagon, pramlintide, human growth hormone, prolactin, mammary trophic hormone, vasopressin, oxytocin, thyroxine, cortisol, etc.), antibodies (including monoclonal and polyclonal antibodies) or fragments thereof, used in the prevention, diagnosis, mitigation, treatment or cure of a condition, illness or disease. (e.g., Fab fragments, Fc fragments, etc.), antibody conjugates (including antibodies or fragments thereof conjugated, i.e., directly or indirectly linked, to another active pharmaceutical ingredient), antibody complexes (e.g., multimeric immunoglobulin complexes), antibiotics, enzymes, and other biologics (e.g., growth factors, colony stimulating factors, interleukins, interferons, etc.), or small molecule active pharmaceutical ingredients (including, but not limited to, anti-cancer small molecule active ingredients, antibiotics, antifungals, anti-inflammatory agents, anticonvulsants, anticoagulants and antithrombotic agents, anti-seizure treatments and prophylactics, anti-migraine treatments and prophylactics, etc.). In certain embodiments, the formulation includes one or more polymer or copolymer carriers that provide sustained release of the therapeutic compound, such as poly(ethylene glycol) ("PEG"), poly(lactic-co-glycolic acid) ("PLGA"), etc. In certain such formulations, the therapeutic agent itself may be complexed or conjugated to one or more such polymers or copolymers.In a further aspect, the formulations of the invention generally include one or more excipients, carriers or buffers, such as one or more sugars (e.g., trehalose, dextrose, sucrose, mannose, fructose, etc.), one or more sugar alcohols (e.g., mannitol, xylitol, glycerol, erythritol, maltitol, sorbitol, etc.), one or more buffers (e.g., histidine, citrate, succinate, lactate, etc.), one or more surfactants (e.g., Span 20, Polysorbate 20, Polysorbate 80, Coliform, etc.), one or more saccharide ... surfactants (e.g., Span 20, Polysorbate 20, Polysorbate 80, Coliform, etc.), one or more saccharides (e.g., trehalose, dextrose, sucrose, mannose, fructose, etc.), one or more sugar alcohols (e.g., mannitol, xylitol, glycerol, ery Miglyol® HS15), triglycerides (e.g., Miglyol® 810, Miglyol® 812, Miglyol® 818, Miglyol® 829, Miglyol® 840), one or more amino acids (which may be any naturally occurring amino acid, such as histidine, proline, glycine, methionine, tryptophan, phenylalanine, arginine, and cysteine), and other pharma- ceutically acceptable carriers, excipients, and fillers that will be familiar to those of skill in the art.

[0021] The formulations provided by the invention are stable and typically do not require reconstitution prior to use, and include concentrated semi-solid or solid formulations of from about 0.1 microliters up to about 3 mL for single dose injectable formulations and up to about 10 mL for infusion formulations, containing an effective amount of at least one therapeutic agent (and in some embodiments, more than one therapeutic agent, e.g., 2, 3, 4 or more therapeutic agents, in mixtures, particularly in combinations where there are two or more therapeutic agents that are not compatible with each other in a typical aqueous formulation) homogenously contained within a pharma- ceutically acceptable carrier. In certain such aspects, the formulation comprises from about 10% to about 95% solids by weight, from about 15% to about 90% solids, or from about 20% to about 85% solids by weight, and in certain preferred embodiments, from about 40% to about 70% by weight, particularly about 40%, about 42%, about 45%, about 50%, about 55%, about 60%, about 65%, about 67% or about 70% by weight. In certain such aspects, the therapeutic or pharma- ceutical active agent has an average particle size ranging from about 10 nanometers (0.01 micrometers) to about 100 micrometers, with no particles larger than about 1 mm, and in certain such embodiments, an average particle size of about 0.1 micrometers to about 25 micrometers, with no particles larger than about 25 micrometers, and in certain other embodiments, an average particle size of about 1 to about 15 micrometers, with at least about half of the particles ranging in size from about 2 micrometers to about 8 micrometers. In particular, the process used to manufacture the present formulations results in formulations in which the particles are relatively uniform in size, although not necessarily considered to be monodisperse. For example, the measured size distribution of the particles (e.g., as measured by standard techniques such as laser diffraction, D 10 , D 50 and D. 90 (reported as D) spans the range of 0.5 to 5.0, or 1.0 to 3.0, or 1.5 to 2.5 (typically reported in the art as D 90 -D 10 ) / D 50Ideally, the particles of the therapeutic agent are of a size and size distribution that promotes high loading efficiency and minimal surface area, properties that can be controlled using the manufacturing processes provided by the invention as described elsewhere herein.

[0022] In certain embodiments, the formulation further comprises one or more carriers (e.g., one or more diluents, additives and / or polymers) that impart thixotropic properties to the formulation. The therapeutic agent is preferably homogeneously incorporated into the pharma- ceutically acceptable carrier, such that the formulation is thixotropic or non-Newtonian in the form of a paste or slurry.

[0023] In certain preferred such embodiments, the therapeutic agent is in powder form and is homogeneously contained in a pharma- ceutically acceptable carrier. The carrier is preferably biocompatible and a non-solvent for the therapeutic agent powder (so that no or minimal dissolution of the powder occurs in the carrier), and in certain preferred embodiments, fills the space between particles of the therapeutic agent powder in a manner that allows the particles to flow. In certain such embodiments, the carrier is selected from the group consisting of alkyl benzoates, aryl benzoates, aralkyl benzoates, triacetin, aprotic polar solvents (e.g., N-methyl-2-pyrrolidine 5 (NMP), dimethyl sulfoxide (DMSO)), medium chain triglycerides (MCTs, e.g., Miglyol® 810, Miglyol® 812N, Miglyol® 818, Miglyol® 829, Miglyol® 840, etc.), alkanes, cyclic alkanes, chlorinated alkanes, fluorinated alkanes, perfluorinated alkanes, and mixtures thereof. The carrier may be a single fluid or semi-solid, or it may be a mixture of two or more fluids (or semi-solids) that may be partially or completely miscible with each other or that are immiscible with each other, such as a mixture of two or more fluids that form an emulsion.

[0024] In certain embodiments, the injectable formulation may provide controlled (delayed) or sustained release. In such embodiments, for example, the formulation may include an effective amount of a pharma- ceutically acceptable polymer to delay the release of the therapeutic agent from the formulation upon administration via injection into the epidermis, dermis or subcutaneous layer of an animal. Agents that facilitate controlled or sustained release of the active pharmaceutical (therapeutic) component in such formulations may be incorporated into the continuous (diluent) phase and / or the disperse (particulate matter) phase of the composition. Additionally or alternatively, the therapeutic agent may be incorporated into liposomes or conjugated or incorporated into polysaccharides and / or other polymers to provide controlled release of the therapeutic agent from the formulation upon administration via injection into the epidermis, dermis or subcutaneous layer of an animal. In certain preferred embodiments, the therapeutic agent may be incorporated into a biocompatible polymer and a poorly water-miscible biocompatible solvent that forms a viscous gel with the polymer and limits water uptake by the composition. Such compositions are disclosed, for example, in U.S. Pat. No. 6,130,200, the entirety of which is incorporated herein by reference, and utilize, for example, a PEG polymer or a PLGA copolymer together with an effective plasticizing amount of a solvent (including, for example, a lower alkyl or aralkyl ester of benzoic acid) to form a gel with the polymer.

[0025] In a further aspect, the present invention also provides a method of parenteral, e.g., intradermal (into the epidermis or dermis), subcutaneous or intramuscular administration of an injectable formulation to an animal (e.g., a human or a veterinary or agricultural animal) to deliver a higher concentration or amount of an active pharmaceutical ingredient to the animal in a smaller amount than would be possible, or to provide painless or substantially painless administration of a therapeutic agent, comprising the step of injecting about 0.1 to about 50 microliters of a concentrated semi-solid or solid formulation (e.g., a slurry or paste) containing about 20 to about 85% solids by weight and containing an effective amount of a therapeutic agent into the epidermal, dermal or subcutaneous skin layer of the animal.

[0026] In preferred embodiments, the therapeutic agent is processed, e.g., via spray drying or freeze drying, to provide a particle size suitable for injection through a narrow gauge needle (e.g., 25-30 gauge). The therapeutic agent is typically processed into a powder with one or more excipients included, e.g., to promote stability, achieve a desired pharmacokinetic profile, and / or improve the ease of manufacture of the therapeutic agent.

[0027] Exemplary processes for producing such formulations are provided elsewhere herein, particularly in the Examples below.

[0028] In certain preferred embodiments, the therapeutic agent is incorporated into a non-aqueous or semi-aqueous pharma- ceutically acceptable carrier, hi further preferred embodiments, the formulation exhibits shear thinning properties upon injection from an injection device.

[0029] The present invention is further directed, in part, to methods of treating animals, eg, human patients or veterinary or agricultural animals, utilizing the injectable formulations, injection devices and preparation methods of the present invention.

[0030] The term "intradermal" includes administration to the skin, ie, the epidermal or dermal skin layers, of an animal, for example a human or veterinary or agricultural animal.

[0031] The term "subcutaneous" refers to administration below the skin layer but above the muscle layer in an animal, such as a human or veterinary or agricultural animal.

[0032] The term "intramuscular" means administration into the muscle layer of an animal, such as a human or a veterinary or agricultural animal.

[0033] The term "pharmaceutically acceptable carrier" refers to a pharmaceutically acceptable solvent, suspending agent, diluent or vehicle for delivering a compound of the present invention to an animal or human. The carrier may be liquid, semi-solid or solid, and may be a Newtonian or non-Newtonian fluid.

[0034] The term "pharmaceutical acceptable" ingredient, excipient or component is one that is suitable for use in humans and / or animals without (or with reduced) undue adverse side effects (such as toxicity, irritation, and allergic reactions) commensurate with a reasonable benefit / risk ratio.

[0035] The term "therapeutic agent" means an agent that, alone or in combination with other pharmaceutical excipients or inactive ingredients, provides a desired, beneficial, often pharmacological, effect upon administration to a human or animal.

[0036] The term "chemical stability" means, with respect to a therapeutic agent, that the proportion of degradation products generated by chemical pathways such as oxidation or hydrolysis is within an acceptable range. In particular, a formulation is considered chemically stable if, after one year of storage at the intended storage temperature of the product (e.g., room temperature); or one year of storage of the product at 30°C / 60% relative humidity; or one month, preferably 3-6 months of storage of the product at 40°C / 75% relative humidity, a maximum of about 50% or less of degradation products are formed, e.g., about 10%, about 20%, about 30%, about 40% or about 50% or less.

[0037] The term "physical stability" means, with respect to a therapeutic agent, that the percentage of aggregates formed (e.g., dimers, trimers and larger forms) is within an acceptable range. In particular, a formulation is considered physically stable if no more than about 15% aggregates are formed after one year of storage at the intended storage temperature of the product (e.g., room temperature); or one year of storage of the product at 30°C / 60% relative humidity; or one month, preferably 3-6 months of storage of the product at 40°C / 75% relative humidity.

[0038] The term "stable formulation" means that at least about 65% of the chemically and physically stable therapeutic agent remains after storage at room temperature for two months. Particularly preferred formulations are those that retain at least about 80% of the chemically and physically stable therapeutic agent under these conditions. Particularly preferred stable formulations are those that do not exhibit degradation after sterilizing irradiation (e.g., gamma, beta, or electron beam).

[0039] The term "bioavailability," for purposes of the present invention, is defined as the extent to which a therapeutic agent is absorbed from a formulation into the bloodstream and / or tissues of an animal or human to which the formulation is administered.

[0040] The term "systemic" means, with respect to delivery or administration of a beneficial agent to a subject, that the beneficial agent is detectable at biologically significant levels in the plasma of the subject.

[0041] The term "paste" refers to a concentrate of a therapeutic agent dispersed in a pharma- ceutically acceptable carrier having a thick consistency to form a viscous, injectable semi-solid. Pastes may be classified as two-phase systems with particulate matter (i.e., solid phase) comprising the "dispersed phase" and a diluent (i.e., non-solvent) comprising the "continuous phase."

[0042] The term "slurry" means a thin paste.

[0043] The terms "controlled release" and "sustained release" are defined for purposes of this invention as the release of a therapeutic agent at a rate such that blood (e.g., plasma) concentrations are maintained within the therapeutic range but below toxic concentrations for a period of about 1 hour or more, preferably 12 hours or more.

[0044] In certain aspects, the invention provides a syringe pre-filled with the high concentration / high viscosity pharmaceutical paste formulation of the invention. In certain such embodiments, the pre-filled syringe comprises a syringe body defining a reservoir and a viscous liquid (e.g., a 250-3000 mg / mL, ... The syringe body includes a paste having a solids concentration of about 675 mg / mL, about 700 mg / mL, about 750 mg / mL, about 800 mg / mL, about 850 mg / mL, about 900 mg / mL, about 950 mg / mL, and about 1000 mg / mL, most specifically about 300 mg / mL to about 850 mg / mL, a plunger and / or piston disposed within the reservoir and configured to move to expel the paste from the reservoir, a luer fitting disposed in the syringe body and in fluid communication with the reservoir, and a closure cap disposed on the luer fitting to seal the reservoir. Some embodiments include a needle defining a lumen, the needle configured to be coupled to the syringe body via the luer fitting to enable intradermal delivery of the paste, and the reservoir having an interior first lateral dimension greater than an interior second lateral dimension of the lumen. This embodiment of the pre-filled syringe may have a needle secured to the syringe via a Luer lock or Luer slip ("slip tip") fitting. Alternative embodiments of the invention may have a needle permanently secured to the syringe body using, for example, a fixed needle configuration in which the needle cannot be removed from the syringe body as with a Luer fitting.

[0045] In certain embodiments, the pre-filled syringe includes a syringe body defining a reservoir having an internal first lateral dimension, a paste disposed within the reservoir and having a solids concentration of at least about 300-600 mg / mL, about 300-600 mg / mL, or greater than about 300-600 mg / mL, a needle defining a lumen having an internal second lateral dimension smaller than the first lateral dimension and configured to be in fluid communication with the reservoir to enable intradermal delivery of the paste, and a plunger disposed within the reservoir and configured to move to expel the paste from the reservoir through the lumen.

[0046] In some embodiments of this pre-filled syringe, the paste has a volume of 15, 40, 50, 100, 150, 250, or 500 μL to 1000, 2000, or 3000 μL. In certain aspects, the paste may have a volume of 15 μL to 1000 μL. In some embodiments, the paste has a volume of up to about 40 μL. In some embodiments, the paste has a volume of up to about 50 μL. In some embodiments, the paste has a volume of up to about 100 μL or up to about 150 μL. In some embodiments, the paste has a volume of up to about 200 μL to about 1000 μL, e.g., about 200 μL, about 300 μL, about 350 μL, about 400 μL, about 450 μL, about 500 μL, about 550 μL, about 600 μL, about 650 μL, about 700 μL, about 750 μL, about 800 μL, about 850 μL, about 900 μL, about 950 μL, or about 1000 μL.

[0047] Some embodiments of the pre-filled syringe are configured to eject paste at a flow rate of at least about 15, about 15, or greater than 15 microliters per second (μL / s) under a force applied to the plunger having a magnitude of about 50, 60, or 70, or at most 50, 60, or 70 Newtons (N). In certain embodiments, the force applied to the plunger may be less than 5, 10, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, or 70 N. In further embodiments, the force applied to the plunger may be less than 25 N. Some embodiments are configured to eject paste at a flow rate of greater than 65 μL / s under a force applied to the plunger having a magnitude of about 50-70 N, or at most 50-70 N. In another aspect, particularly those using an auto-injector or other auxiliary delivery device (e.g., a reusable auto-injector), the pre-filled syringe or device is configured to eject the paste at a flow rate of the above forces, or at a force of greater than about 70 N, e.g., about 75 N, about 80 N, about 85 N, about 90 N, about 95 N or about 100 N.

[0048] Some embodiments of the kit include a syringe body defining a reservoir having an internal first transverse dimension, and a needle configured to be coupled to the syringe body and defining a lumen having an internal second transverse dimension smaller than the first transverse dimension. In some embodiments, the paste is disposed in the reservoir. In some embodiments, the syringe body includes a luer fitting (e.g., a luer lock or luer slip fitting) in communication with the reservoir, and a closure cap disposed on the luer fitting to seal the reservoir, and the needle is configured to be coupled to the syringe body via the luer fitting. In another embodiment, the needle is integral with the syringe body without being removably connected. In some embodiments, the reservoir has a volume of 50, 75, or 100 μL to 1000, 2000, or 3000 μL.

[0049] Some embodiments of the kit include a plunger disposed within the reservoir and configured to move to expel the paste from the reservoir through the lumen at a flow rate of greater than 30 μL / s under a force applied to the plunger of a magnitude as described elsewhere herein. Some embodiments include a plunger disposed within the reservoir and configured to move to expel the paste from the reservoir through the lumen at a flow rate of greater than 65 μL / s under a force applied to the plunger of a magnitude as described elsewhere herein.

[0050] An alternative embodiment is the use of bolus injectors, also known as patch pumps or large volume injectors. In certain aspects, patch pumps can be employed for long-term delivery of viscous pastes to patients. Examples of these injectors include the SmartDose™ electronic wearable bolus injector (West Pharmaceutical Services, Inc.) and the Lapas® bolus injector (Bespak), as well as others known in the art (see, e.g., Badkar AV et al., Drug Des. Devel. Ther. 15: 159-170 (2021), doi:10.2147 / DDDT.S287323). These devices can be worn on the body and can provide automatic subcutaneous or intradermal delivery of highly concentrated pastes at slower injection rates than traditional auto-injectors or manually operated syringes. In these devices, the paste is loaded into an internal reservoir and slowly injected into the patient at a low volumetric flow rate (compared to manual syringes and auto-injectors). These devices can be worn like a patch applied to the skin and deliver the drug for a few minutes or up to about an hour. As a non-limiting example of the volumetric flow rates that may be employed with these systems, delivery of 3 mL of therapeutic paste in 10 minutes would require a delivery rate of 5 μL / sec. Delivery of a volume of paste of 3 mL in 1 hour would require a delivery rate of 0.83 μL / sec.

[0051] Some embodiments of the present method for intradermal injection of a quantity of paste include displacing a plunger of a syringe to expel the paste from a reservoir of the syringe through a lumen of a needle of the syringe, the reservoir having an internal first transverse dimension greater than an internal second transverse dimension of the lumen, e.g., the second transverse dimension being 0.1-0.9 mm, the paste having a solids content of about 20% to about 80% (including all values ​​and ranges therebetween) and a solids concentration of greater than about 100 mg / mL, e.g., about 300 to about 800 mg / mL (including all values ​​and ranges therebetween), particularly a concentration of active pharmaceutical ingredient of about 300 to about 600 mg / mL (including all values ​​and ranges therebetween), and the paste being expelled at a flow rate of greater than 30 μL / s as the plunger is displaced at a rate of 0.5-50 millimeters per second (mm / s). Some embodiments include placing a needle into and / or through the skin tissue of the patient. Some embodiments include removing the sealing cap from a luer fitting of the reservoir. Some embodiments include coupling the needle to the reservoir via a luer fitting disposed on at least one of the needle and the reservoir. In some embodiments, the flow rate of the paste is substantially linearly proportional to the speed of movement of the plunger.

[0052] In some embodiments of the method, the paste injection volume is greater than about 1 μL. In some embodiments, the paste injection volume is from 15, 30, or 100 μL to 1200, 2000, or 3000 μL for a single injection dose, and up to about 10 mL for infusion applications. In some embodiments of the syringe, kit, and / or method, the first lateral dimension is greater than the second lateral dimension. In some embodiments, the first lateral dimension is 1, 2, 3, 4 to 5, 6, 7, 8, 9, 10, 11, 12 mm (including all values ​​and ranges therebetween). In some embodiments, the second lateral dimension is 0.1, 0.2, 0.3, or 0.4 to 0.5, 0.6, 07, 0.8, or 0.9 mm (including all values ​​and ranges therebetween).

[0053] In some embodiments of the present syringes, kits, and / or methods, the needle is 18 gauge or larger in size (where larger gauge refers to a physically smaller needle in terms of outer and / or inner diameter of the needle). In some embodiments, the needle is 23 gauge or smaller in size. In some embodiments, the needle is 25 gauge or 27G or smaller in size (i.e., larger gauge). In some embodiments, the needle has an exposed length of about 50 mm or less. In some embodiments, the needle has an exposed length of about 40 mm or less. In some embodiments, the needle has an exposed length of about 13 mm or less. In some embodiments, the needle has an exposed length of approximately 8 mm. In some embodiments, the needle has an exposed length of approximately 6 mm.

[0054] In some embodiments of the present syringes, kits, and / or methods, the paste has a solids concentration of greater than 200 mg / mL. In some embodiments, the paste has a solids concentration of 200-800 mg / mL. In some embodiments, the paste has a solids concentration of 300-750 mg / mL. In some embodiments, the paste has a solids content of 1%-99%. In some embodiments, the paste has a solids content of 30%-75%. In some embodiments, the paste has a solids content of 40%-65% or 50%-60%. In some embodiments, the paste has a density of about 0.5, 0.7, 0.75, 1.0, 1.1, 1.2, 1.3 to about 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, 2.0 g / mL (including all values ​​and ranges therebetween).

[0055] As used in this disclosure, a paste is a two-phase mixture of a solid (e.g., a powder containing a drug and optionally a stabilizer and / or excipient) dispersed in a liquid (e.g., a biocompatible diluent) that is a non-solvent for the solid or only slightly solubilizes it (e.g., and therefore the diluent is typically, but not always, lipophilic in nature). A paste behaves as a solid until a sufficiently large load or stress (typically referred to as "yield stress") is applied, at which point the paste flows like a liquid (e.g., a paste may be defined as a semi-solid). A paste may exhibit non-Newtonian fluid behavior, particularly shear-thinning properties and / or viscoelastic behavior.

[0056] The term "coupled" is defined as connected, although not necessarily directly, and not necessarily mechanically, such that two items that are "coupled" may be integral with one another.

[0057] The terms "a" and "an" are defined as one or more, unless the disclosure expressly states otherwise.

[0058] The term "substantially" is defined as being largely, but not necessarily all, of what is specified (and includes what is specified, e.g., substantially 90 degrees includes 90 degrees, substantially parallel includes parallel), as will be understood by one of ordinary skill in the art. In any disclosed embodiment, the terms "substantially," "approximately," and "about" may be substituted for "within [percent]" of what is specified, where percent includes .1, 1, 5, 10, and 20 percent.

[0059] As used herein, the term "intracutaneous injection" includes epidermal, intradermal, subcutaneous or intramuscular injections.

[0060] As used herein, a "phase" is defined as a homogenous, physically distinct portion of a system that is separated from the rest of the system by an interface. Three major phases of matter are known to exist: solid, liquid, and gas. As an example, a system containing particulate matter suspended in a liquid that is a non-solvent for the particulate matter is considered to be a two-phase system. Conversely, a system consisting of organic macromolecules uniformly distributed throughout the liquid such that there is no clear boundary between the macromolecules and the liquid molecules is considered to be a single-phase solution.

[0061] As used herein, "semi-solid" is an attribute of a material that exhibits plastic flow behavior. A semi-solid material is not pourable, does not easily conform to its container at room temperature, and does not flow at low shear stresses. Thus, a semi-solid has a yield stress that must be overcome before plastic (i.e., irreversible) deformation occurs. Semi-solids typically have a viscoelastic rheological flow profile.

[0062] Thus, semi-solid refers not to a specific physical composition or pharmaceutical dosage form, but rather to the physical characteristics of a material. Thus, various materials can be considered semi-solid because they have the attributes of a semi-solid material, despite being physically distinct compositions. For example, the USP-NF describes both creams and medicated foams as having a semi-solid consistency, and thus, both can be considered semi-solid fluids or semi-solids, despite being otherwise physically distinct compositions. Similarly, gels and pastes are often both referred to as semi-solids, despite being physically distinct. Gels are defined by the USP-NF as a dosage form that is a semi-solid dispersion of small particles or a solution of large molecules interpenetrated with a solution containing a gelling agent that provides rigidity. Thus, gels can be either single-phase or two-phase systems. Remington: The Science and Practice of Pharmacy(2006), gel systems can be either transparent or opaque since the components that make up the gel may not be completely soluble or insoluble, or may form aggregates and scatter light. Gels are defined as "semi-rigid systems in which the motion of the dispersion medium is restricted by an entangled three-dimensional network of particles or solvated macromolecules in the dispersed phase...the entanglement and resulting internal friction cause an increased viscosity and a semi-solid state."

[0063] Gels in which the polymers are dispersed throughout the liquid in a manner that does not present a clear boundary between the polymers and the liquid are called single-phase gels. When the gel mass consists of aggregates of small, separate particles, the gel is classified as a two-phase system and is often called magma or milk. Gels and magmas each contain particles of colloidal dimensions and are therefore considered colloidal dispersions. The generally accepted size range for "colloidal" materials is when the particles are in the range of 1 nm to 0.5 μm.

[0064] In contrast, a paste may be defined as a semi-solid dosage form containing a high percentage of finely dispersed solids with a stiff consistency. As mentioned above, the actual solids content of a paste will vary primarily depending on the properties of the constituent powders. To prepare a paste, the minimum amount of fluid added to the powder must be sufficient to coat and produce a monolayer of fluid around each individual powder particle. It should be noted that this is an ideal situation where all powder-powder contact is completely blocked, and in reality, many finely divided powders are highly cohesive and it may not be possible to completely block all direct powder-powder contact, even when high shear mixing techniques are applied. Additional fluid is then added to the mixture to fill the gaps (i.e., void volume) between the powder particles, allowing the particles to flow as a fluid when the yield stress of the paste is exceeded. Thus, powders with very low density (i.e., high surface area to volume ratio) and / or poor packing (i.e., large gaps between particles) require a larger volume / mass of fluid to form a paste than powders with low surface area to volume ratios and / or good packing properties. Thus, although both gels and pastes may have semi-solid properties and may both be referred to as semi-solids, they are physically different dosage forms. In particular, the solids concentration of pastes is typically much higher, and the particles are often much larger than the upper limit of the colloidal region (0.5 μm). Overall, USP-NF defines at least six different dosage forms as semi-solid, including creams, foams, gels, jellies, ointments, and pastes. However, it will be easily recognized and understood by those skilled in the art that, although these pharmaceutical dosage forms are different physical compositions with different physical properties, they all have the rheological properties of semi-solids, and are therefore broadly referred to as semi-solids.

[0065] "Solid content" as used herein refers to the weight percentage (wt%) of the solid phase (e.g., powder) in a paste as a percentage of the total mass of the two phases (solid plus liquid) that make up the paste. Solid content is typically expressed / discussed in terms of %. As an example, if 1 g of paste is prepared by blending 0.65 g of the solid phase and 0.35 g of the liquid phase, the solid content of the paste is 65%.

[0066] "Solids concentration" as used herein refers to the mass of solid phase per unit volume of paste. Typical units of solids concentration include mg / mL and g / mL. The solids concentration of a paste is related to the solids content and can be obtained by multiplying the solids content of the paste by the density of the paste (measured using a suitable method such as helium pycnometer). For example, a paste with a density of 1250 mg / mL (1.25 g / mL) and a solids content of 60% would have a solids concentration of approximately 750 mg / mL (0.75 g / mL). It should be noted that the drug concentration in the solid phase of the paste can be equal to or less than the solids concentration of the paste, and is typically lower than the solids concentration due to the presence of additional components (e.g., bulking or stabilizing excipients) in the solid phase that dilute the drug concentration.

[0067] "Non-Newtonian fluid," as used herein, defines a fluid whose viscosity varies with shear rate or shear rate history, as opposed to a Newtonian fluid, whose viscosity is typically independent of the applied shear rate.

[0068] "Thixotropic," as used herein, defines a fluid that exhibits shear-thinning properties. More specifically, thixotropic fluids exhibit time-dependent shear-thinning properties, as opposed to pseudoplastic fluids, which may be characterized as fluids that exhibit time-independent shear-thinning properties. However, for purposes of this application, thixotropic fluids generally describe shear-thinning fluids.

[0069] The term "pharmaceutical acceptable" as used herein means suitable for normal pharmaceutical use, i.e. causing no serious adverse events in patients.

[0070] The term "pharmaceutically acceptable carrier" refers to a pharmaceutically acceptable solvent, suspending agent or vehicle for delivering a compound of the invention to an animal or human. The carrier may be liquid, semi-solid or solid.

[0071] The term "pharmaceutical acceptable" ingredient, excipient or component is one that is suitable for use in humans and / or animals without undue adverse side effects (such as toxicity, irritation, and allergic reactions) commensurate with a reasonable benefit / risk ratio.

[0072] The term "therapeutic agent," which is used interchangeably herein with the terms "pharmaceutical active ingredient," "active ingredient," or "active pharmaceutical ingredient," means an agent that, alone or in combination with other pharmaceutical excipients or inactive ingredients, provides a desired, beneficial, and often pharmacological, effect upon administration to a human or animal. In certain aspects of the invention, therapeutic agents include drugs (e.g., small molecules, peptides, proteins, biologics), vaccines, oligonucleotides, gene therapy vehicles / vectors, and the like, used in the prevention, diagnosis, mitigation, treatment, or cure of a condition, illness, or disease.

[0073] The term "chemical stability" means, with respect to a therapeutic agent, that the percentage of degradation products generated by chemical pathways such as oxidation or hydrolysis is within an acceptable range. In particular, a formulation is considered chemically stable if no more than about 20% degradation products are formed after one year of storage at the intended storage temperature of the product (e.g., 4°C (refrigerated) or 25°C (room temperature)); or one year of storage of the product at 30°C / 60% relative humidity; or one month, preferably 3-6 months of storage of the product at 40°C / 75% relative humidity.

[0074] The term "physical stability" means, with respect to a therapeutic agent, that the percentage of aggregates formed (e.g., dimers, trimers and larger forms) is within an acceptable range. In particular, a formulation is considered physically stable if, after one year of storage at the intended storage temperature of the product (e.g., refrigerated or room temperature); or one year of storage of the product at 30°C / 60% relative humidity; or one month, preferably 3-6 months of storage of the product at 40°C / 75% relative humidity, no more than about 15% aggregates are formed, preferably no more than about 1-10% or 1-5%.

[0075] The term "stable formulation" means that at least about 65% of the therapeutic agent remains chemically and physically stable after storage at room temperature for two months. Particularly preferred formulations are those that retain at least about 80% of the therapeutic agent chemically and physically stable under these conditions.

[0076] The term "bioavailability," for purposes of the present invention, is defined as the extent to which a therapeutic agent is absorbed from a formulation.

[0077] The term "systemic" means, with respect to delivery or administration of a beneficial agent to a subject, that the beneficial agent is detectable at biologically significant levels in the plasma of the subject.

[0078] The term "slurry" means a thin paste (the term "paste" is defined below).

[0079] The term "controlled release" is defined for purposes of this invention as the release of a therapeutic agent at a rate such that blood (e.g., plasma) concentrations are maintained within the therapeutic range but below toxic concentrations for a period of about 1 hour or more, preferably 12 hours or more.

[0080] Moreover, a device or system configured in a particular manner may be configured in at least that manner, but may be configured in other manners than those specifically described.

[0081] The terms "comprise" (and all forms of comprise, such as "comprises" and "comprising"), "have" (and all forms of have, such as "has" and "having"), "include" (and all forms of include, such as "includes" and "including"), and "contain" (and all forms of contain, such as "contains" and "containing") are open-ended linking verbs. As a result, an apparatus that "comprises," "has," "includes," or "contains" one or more elements includes those one or more elements, but is not limited to including only those elements. Similarly, a method that "comprises," "has," "includes," or "contains" one or more steps includes those one or more steps, but is not limited to including only those steps.

[0082] Any aspect of any of the devices, systems, and methods may consist of or consist essentially of any of the described steps, elements, and / or features, rather than comprise / include / contain / have. Thus, in any claim, the terms "consisting of" or "consisting essentially of" may be substituted with any of the above open-ended linking verbs to change the scope of a given claim from that which would result from the use of the open-ended linking verb.

[0083] Even if not described or illustrated, a feature or features of one embodiment may be applied to other embodiments unless expressly prohibited by the nature of the disclosure or the embodiment.

[0084] Other objects, advantages, and features of the present invention will become readily apparent to those of ordinary skill in the art upon review of the description, drawings, examples, and claims set forth herein. [Brief description of the drawings]

[0085] The following drawings illustrate by way of example and not limitation. For the sake of brevity and clarity, every feature of a given structure is not always labeled in every drawing in which that structure appears. The same reference numbers do not necessarily refer to the same structures. Rather, the same reference numbers may be used to refer to similar features, or features with similar functionality, while different reference numbers may do so. The drawings are drawn to scale (unless otherwise noted), which means that the dimensions of the depicted elements are accurate relative to each other, at least with respect to the aspects depicted in the drawings. [Figure 1] FIG. 1 is a pair of scanning electron micrographs showing particles of therapeutic protein powder (in this case, monoclonal antibodies (mAbs)) prepared according to the inventive spray drying method using different spray dryer (Buchi B290) equipment settings. FIG. 1A: Inlet temperature 90° C., aspirator 65% (27 m3 / hr), nozzle gas flow rate 473 L / hr (pressure drop 0.41 bar), feed pump flow rate 3% (approximately 1 mL / min), and no secondary drying or processing (e.g., sieving) after spray drying. FIG. 1B: Inlet temperature 70° C., aspirator 85% (34 m3 / hr), nozzle gas flow rate 473 L / hr (pressure drop 0.41 bar), feed pump flow rate 10% (approximately 3 mL / min), and secondary drying (lyophilization) and sieving were performed after spray drying. [Diagram 2] Figure 2 is a series of scanning electron micrographs showing particles of a therapeutic protein powder (in this case, a monoclonal antibody) prepared according to the inventive spray drying method using different spray dryer equipment settings, as shown in Table 4 below: Figure 2A: Formulation 1; Figure 2B: Formulation 2; Figure 2C: Formulation 3; Figure 2D: Formulation 4; Figure 2E: Formulation 5; Figure 2F: Formulation 6; Figure 2G: Formulation 7; Figure 2H: Formulation 8. [Diagram 3]Figure 3 is a series of bar graphs showing particle size distribution (assessed by visual inspection via scanning electron microscope) of therapeutic protein powders (in this case, monoclonal antibodies) prepared according to the inventive spray drying method using different spray dryer apparatus process settings. Formulation numbers and spray dryer settings correspond to those described in the legend to Figure 2 above. Figure 3A: Formulation 1; Figure 3B: Formulation 2; Figure 3C: Formulation 3; Figure 3D: Formulation 4; Figure 3E: Formulation 5; Figure 3F: Formulation 6; Figure 3G: Formulation 7; Figure 3H: Formulation 8. [Figure 4] FIG. 4 is a chart (top) showing the percent aggregation of certain formulations prepared according to the methods of the invention before and after spray drying (at time zero after spray drying, and after storage at 50° C. for 1 day after spray drying), and a bar graph illustrating these results. [Diagram 5] Figure 5 is a chart (top) showing percent aggregation of certain formulations prepared according to the methods of the invention before and after spray drying (at time zero after spray drying, and after storage at 50°C for 1 day after spray drying), and a bar graph illustrating these results. Secondary drying was performed under reduced pressure in a freeze dryer (lyo) after spray drying. [Figure 6] FIG. 6 is a chart (top) showing the percent aggregation of certain formulations prepared according to the methods of the invention before and after spray drying (at time zero after spray drying, and after storage at 50° C. for 1 day after spray drying), as well as a bar graph illustrating these results. [Figure 7] FIG. 7 is a chart showing the percent aggregation of certain formulations prepared according to the methods of the invention before and after spray drying (at time zero (T=0) after spray drying, and after post-drying by storage at 40° C. for 5 days). [Figure 8] FIG. 8 is a chart (top) showing the percent aggregation of certain formulations prepared according to the methods of the invention before and after spray drying (at time zero after spray drying, and after storage at 50° C. for 1 day after spray drying), and a bar graph illustrating these results. [Figure 9]FIG. 9 is a chart (top) showing the percent aggregation of certain formulations prepared according to the methods of the invention before and after spray drying (at time zero after spray drying, and after storage at 50° C. for 1 day after spray drying), as well as a bar graph illustrating these results. [Figure 10] FIG. 10 is a chart (top) showing the percent aggregation of certain formulations prepared according to the methods of the invention before and after spray drying (at time zero after spray drying, and after storage at 50° C. for 1 day after spray drying), as well as a bar graph illustrating these results. [Figure 11] FIG. 11 is a chart (top) showing the percent aggregation of certain formulations prepared according to the methods of the invention before and after spray drying (at time zero after spray drying, and after storage at 50° C. for 1 day after spray drying), as well as a bar graph illustrating these results. [Figure 12] FIG. 12 is a chart (top) showing the percent aggregation of certain formulations prepared according to the methods of the invention before and after spray drying (at time zero after spray drying, and after storage at 50° C. for 1 day after spray drying), as well as a bar graph illustrating these results. [Figure 13] FIG. 13 is a series of charts showing the main peak ( FIG. 13A ), acidic variant ( FIG. 13B ), and basic variant ( FIG. 13C ) observed upon ion exchange chromatography of a cysteine-containing formulation prepared according to the methods of the invention before ("pre-SD") and after (time zero (t0) after spray drying and after storage at 50° C. for 1 day after spray drying (50C×1d)). [Figure 14] FIG. 14 is a comparison of representative traces of two formulations, one containing 1.5 mg / mL cysteine ​​(FIG. 14A) and the other containing 6 mg / mL cysteine ​​(FIG. 14B), prepared according to the methods of the invention before spray drying ("Pre-SD") and after (time zero after spray drying ("t0") and after storage at 50° C. for 1 day after spray drying ("50C×1d")). [Figure 15]FIG. 15 is a chart (top) showing the percent aggregation of certain formulations prepared according to the methods of the invention before and after spray drying at different inlet temperatures (at time zero after spray drying, and after storage at 50° C. for 1 day after spray drying), and a bar graph illustrating these results. [Figure 16] FIG. 16 is a series of scanning electron micrographs showing particles of therapeutic peptide powders (in this case monoclonal antibodies) spray dried from commercial formulations, showing particles observed in spray dried formulations of Herceptin (TmAb) (FIG. 16A), Erbitux (cetuximab) (FIG. 16B), and Pirivigen (immunoglobulin) (FIG. 16C). [Figure 17] FIG. 17 is a chart showing particle size distribution in the three formulations shown in FIG. 16, measured using laser diffraction. D10: 10th percentile particle size; D50: 50th percentile particle size; D90: 90th percentile particle size. Span=(D90-D10 / D50). [Figure 18] FIG. 18 is a line graph showing the injection force (measured using a texture analyzer) required to expel 1 mL of an inventive paste formulation using a 1 mL long syringe equipped with a 23 G needle (lower trace) or a 27 G needle (upper trace). [Figure 19] FIG. 19 is an ion exchange chromatogram showing peaks obtained with formulations of trastuzumab (Herceptin®) in commercial aqueous form (red trace), as a powder prepared by the inventive spray drying method and then reconstituted with water (green trace), or as an inventive XeriJect™ paste formulation (pink trace). [Figure 20] FIG. 20 is a line graph (top) showing the pharmacokinetics (PK) of Trastuzumab (Herceptin®) formulations injected into test animals and evaluated for plasma antibody concentrations, the commercial aqueous form (blue trace) injected intravenously into test animals, as well as two inventive Xeriject™ paste formulations, one using a 10 mg / Kg dose and the other using a 20 mg / Kg dose, injected subcutaneously (FIG. 20A); and a chart showing certain PK parameters in tabular form (FIG. 20B). [Figure 21] FIG. 21 is a pair of scanning electron micrographs of human enzyme preparation powders prepared by lyophilization from a commercial aqueous formulation (FIG. 21A) or by the spray drying method of the present invention (FIG. 21B). [Figure 22] FIG. 22 is a series of line graphs showing the pharmacokinetic (PK) results of intravenous injection of the commercially available aqueous formulation of the enzyme used in FIG. 21 (FIG. 22A), or subcutaneous injection of the aqueous enzyme formulation (FIG. 22B, "Group 2")) or the Xeriject™ paste of the enzyme prepared according to the methods of the present invention (FIG. 22B, "Group 3"). [Diagram 23] FIG. 23 is a series of line graphs showing the pharmacodynamic results of intravenous injection of the commercially available aqueous formulation of the enzyme used in FIG. 21 (FIG. 23A), or subcutaneous injection of the aqueous enzyme formulation (FIG. 23B, "Group 2")) or the Xeriject™ paste of the enzyme prepared according to the methods of the present invention (FIG. 23B, "Group 3"). [Figure 24] FIG. 24 is a series of line graphs showing the pharmacokinetic (FIG. 24A) and pharmacodynamic (FIG. 24B) results of subcutaneous injection of a commercially available aqueous glucagon formulation ("GEK" in FIGS. 24A and 24B) or Zelliject™ Paste of glucagon prepared according to the methods of the present invention ("Zellis Paste" in FIGS. 24A and 24B). [Diagram 25] Figure 25 is a pair of scanning electron micrographs of human recombinant protein formulation powders prepared by the spray drying method of the present invention from a commercially available aqueous formulation. Figure 25A: low concentration feed; Figure 25B: high concentration feed. [Figure 26] FIG. 26 is a bar graph showing the injection force (measured using a texture analyzer) required to inject approximately 150 μL of recombinant protein paste prepared from the powder shown in FIG. 25 at a volumetric flow rate of 30 μL per second using commercially available large and small syringes fitted with either standard or thin walled 27 G needles. [Figure 27] Figure 27 is a pair of scanning electron micrographs of human monoclonal antibody (Bevacizumab, BmAb) formulation powders prepared by the spray drying method of the present invention: Figure 27A: Formulation XJ-1 (pH 4.0); Figure 27B: Formulation XJ-2 (pH 6.0). [Figure 28] Figure 28 is a pair of pharmacokinetic line graphs showing plasma concentrations over time of various formulations of Xeriject bevacizumab (BmAb) after injection into minipigs. Figure 28A: Linear scale; Figure 28B: Same results but semi-logarithmic scale. [Figure 29] FIG. 29 is a bar graph showing the time to maximum plasma concentration (Tmax) for various formulations of Xeriject bevacizumab following injection into minipigs. [Diagram 30] FIG. 30 is a pair of bar graphs showing maximum plasma concentrations (Cmax) for various formulations of Xeriject bevacizumab following injection into minipigs, uncorrected (FIG. 30A) or corrected (FIG. 30B) for dose. [Diagram 31] FIG. 31 is a bar graph showing the plasma half-life (T1 / 2) for various formulations of Xeriject bevacizumab following injection into minipigs. [Diagram 32] Figure 32 is a pair of bar graphs showing dose-corrected total animal exposure for various formulations of Xeriject bevacizumab following injection into minipigs. Figure 32A: dose-corrected AUClast; Figure 32B: dose-corrected AUC∞. [Diagram 33] FIG. 33 is a bar graph showing dose-adjusted fractional animal exposure (AUC336) 14 days after injection for various formulations of bevacizumab following injection into minipigs. [Diagram 34] FIG. 34 is a line graph showing the mean (±SEM) plasma insulin concentrations following subcutaneous administration of Humulin R and Xeriject insulin formulations in Yucatan minipigs. [Diagram 35] FIG. 35 is a line graph showing the mean (±SEM) blood glucose concentrations following subcutaneous administration of Humulin R and Xeriject insulin formulations in Yucatan minipigs. [Diagram 36] Figure 36 is a pair of scanning electron micrographs of an exemplary spray-dried IgG powder formulation produced by the methods of the invention. Images are provided at different magnifications. Two different magnifications are shown: high (Figure 36A; scale bar = 10 μm) and low (Figure 36B; scale bar = 20 μm) magnification. [Figure 37] FIG. 37 is a line graph showing particle size distribution analysis of an exemplary spray dried IgG powder formulation produced by the methods of the invention. [Figure 38] Figure 38 is a series of scanning electron micrographs of an exemplary spray-dried IgG paste produced by the methods of the invention. Images are provided at different magnifications. Three different magnifications are shown: high (Figure 38A; scale bar = 10 μm and 38B; scale bar = 8 μm) and low (Figure 38C; scale bar = 20 μm) magnification. [Figure 39] Figure 39 is a line graph showing particle size distribution analysis of an exemplary spray dried IgG powder formulation produced by the methods of the invention. X-axis: distance (mm); Y-axis: force (N). DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0086] Detailed Description of the Invention Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by those skilled in the art to which this invention belongs. Although any methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present invention, the preferred methods and materials are described below.

[0087] Previous work by several of the present inventors described methods for producing formulations prepared in the form of pastes with high solids content that can contain relatively high concentrations of pharma- ceutically active compounds (see, for example, U.S. Patent Nos. 8,790,679, 8,110,209, and 9,314,424, and U.S. Patent Application Publication Nos. 2017 / 0007675 and 2017 / 0216529, all of whose disclosures are incorporated herein by reference in their entirety). The present invention represents a dramatic development and improvement of this previous work, resulting in formulations that not only have a higher solids content and thus can contain higher concentrations of active ingredients (resulting in reduced injection volumes and easier control of the pharmacokinetic and pharmacodynamic properties of the active substance upon delivery to the patient, especially in the case of subcutaneous injection into the patient), but can also be storage stable for extended periods of time. Furthermore, the methods provided by the present invention allow for the production of formulations containing active ingredients with higher molecular weights and / or greater solids contents than previously believed possible and thus previously demonstrated.

[0088] The present invention has at least four elements that result in the administration of a concentrated therapeutic agent to the intradermal space of a patient, which, in the case of a vaccine or targeted therapeutic product, results in exposure of the antigen to the systemic circulation of the drug or to the immune system. These elements are: (i) a small injection volume compared to the large injection volumes required with the corresponding (typically aqueous) solutions prepared at significantly lower therapeutic agent concentrations; (ii) a concentrated therapeutic agent (e.g., drug) particle or dispersed solid formulation surrounded by a protective solution (typically non-aqueous); (iii) a small diameter needle suitable for intradermal administration; and (iv) a shallow injection of a concentrated dispersion (e.g., paste, slurry) of the therapeutic agent into the epidermal, dermal, subcutaneous, or intramuscular layers of the skin. A composition that meets all of these elements is provided by the manufacturing method of the present invention, which results in an improvement in both the achievable solids loading and the storage stability of the resulting paste formulation when compared to previous compositions and methods.

[0089] Compositions and methods for making same In a first aspect, the present invention provides a method for producing or manufacturing a high solids content and viscous paste containing one or more active pharmaceutical ingredients that can be delivered through a typical needle used for intradermal and / or intramuscular injection, and a composition produced by such a method. In certain such aspects, the present invention provides a method for use in the preparation of a high solids content paste formulation that, when an aqueous formulation is dried into a powder formulation and mixed with a non-aqueous diluent, can have a relatively high active ingredient concentration and is suitable for administration to an animal (e.g., a human or veterinary animal) in a relatively small amount that delivers a therapeutic bolus of one or more active pharmaceutical ingredients in a manner that minimizes the discomfort and / or injection reaction experienced by the animal after injection. Previous such methods have utilized certain freeze-drying, spray-drying, and other particle engineering approaches to prepare the powder used in making the paste formulation, but the inventors have found that such methods have undesirable upper limits on the solids content and active ingredient concentration of the paste. Thus, one objective of the present invention was to identify a suitable manufacturing method that allows for the formation of paste formulations with higher solids concentrations and increased active agent content that can be delivered intradermally and / or intramuscularly in relatively small amounts using commercially available syringe / needle combinations without clogging the needle or requiring the use of excessive injection forces and / or delivery times that often cause discomfort to the animal receiving the therapeutic agent.

[0090] Thus, in one aspect, the present invention provides a method for preparing a paste formulation suitable for use according to the object of the invention. The inventors have discovered that the preparation of a high solids paste requires the preparation of a starting powder material containing both an active pharmaceutical ingredient and one or more pharma- ceutically acceptable excipients or carriers in a manner that results in the formation of powder particles that are preferably spherical and have a preferred size and size distribution (characterized using standard techniques such as laser diffraction). As described in the examples herein, such powders can advantageously form pastes with higher solids concentrations (potentially higher therapeutic agent loadings) than have been available so far.

[0091] Suitable manufacturing methods for making such powders are described in detail in Examples 1 and 2 below. Briefly, an aqueous formulation of one or more active pharmaceutical ingredients is buffer exchanged (e.g., at 4-25°C) against a suitable solution to facilitate protein concentration and / or buffer exchange. Such suitable solutions may include one or more sugars (e.g., trehalose or sucrose), one or more buffering and / or stabilizing agents (e.g., lactate, citrate, succinate, histidine, phosphate, glycine, arginine, proline, methionine, etc.), one or more surfactants / surface active agents (e.g., polysorbate 20, polysorbate 80), etc. Passive and / or active methods for facilitating protein concentration and / or buffer exchange can be used, including but not limited to dialysis, filtration, and centrifugation. For example, aqueous formulations may be prepared using tangential flow filtration (TFF) at room temperature followed by adding one or more sugars (e.g., trehalose or sucrose), one or more buffering and / or stabilizing agents (e.g., lactate, citrate, succinate, glycine, proline, histidine, arginine, methionine, etc.), one or more surfactants (e.g., polysorbate 20, polysorbate 80), etc. directly to the solution. Following concentration and / or buffer exchange using a suitable method (e.g., dialysis, TFF, etc.), the aqueous formulation is subjected to a two-stage drying: (1) spray drying the powder using certain settings of the spray drying apparatus (e.g., BUCHI B-290 Mini Spray Dryer) - i.e., an inlet temperature of about 70°C to about 90°C, preferably about 70°C to 80°C; a drying gas flow rate of about 40-60 mm (equivalent to approximately 470-800 L / hr) measured with a B-290 ball flow meter; an aspirator flow rate of 70% to 100% of the B-290 control (approximately 30-40 mm); 3 / hr), preferably about 85%, 90%, 95% or 100%; a feed flow rate of about 3-20% (corresponding to 1-6 mL / min) relative to the B-290 control; and (2) drying the spray-dried powder under reduced pressure (vacuum) for a suitable time to reduce the moisture content of the powder to a specified level (e.g., less than 5% (w / w), less than 4% (w / w), less than 3% (w / w), less than 2% (w / w) or less than 1% (w / w)). Subsequent drying steps after spray drying are sometimes referred to as secondary drying steps and can be carried out using a variety of techniques known in the art (e.g., under reduced pressure at ambient or other temperatures, under a continuous flow of inert gas at ambient or other temperatures, etc.).

[0092] After production, the resulting powders are treated (e.g., sieved, milled, etc.) if desired to reduce agglomerates present in the bulk solid phase. Using this approach, we have observed a more uniform spherical morphology and generally polydispersity (e.g., polydispersity is measured by the particle size span (D 90 -D 10 / D 50 In an effort to produce a powder starting material having a particle size distribution of 0.1 μm to 1.00 μm, the powder starting material was successfully prepared, which appears to have a particle size distribution of 0.1 μm to 1.00 μm, resulting in a more suitable starting material for the preparation of high solids paste formulations of the invention, as discussed in the Examples below.

[0093] A paste can generally be described as a two-phase composition in which a solid phase (e.g., particulate matter, powder) is mixed with a liquid phase (e.g., diluent) and the solid phase is generally insoluble or at least not completely soluble in the liquid phase. In such a mixture, the liquid phase may be referred to as the continuous phase and the solid phase as the dispersed phase. As will be understood by those skilled in the art, the terms "continuous phase" and "dispersed phase" may also be used to describe compositions prepared from mixtures having similar phases, for example, from two or more liquids that are not completely miscible with each other, including, as non-limiting examples, oil-in-water (O / W) and water-in-oil-in-water (W / O / W) emulsions. A two-phase composition is different from a single-phase composition, such as a solution, which is generally described as a preparation containing one or more chemicals dissolved in a suitable solvent or in a mixture of mutually miscible solvents.

[0094] Broadly, pastes can be defined as multicomponent formulations that fall on the spectrum between solutions and wet solids. In solutions, the solids concentration is low enough that particles eventually begin to settle (e.g., under gravity) when the vial sits undisturbed for shelf-life relevant to commercial pharmaceutical products (e.g., 1 month, 6 months, 12 months, 18 months, 24 months). At the opposite end of the spectrum are wet solids, where there is an excess of solid phase relative to the liquid phase, resulting in a texture that can be broadly described as wet sand, loam, silt, and / or clay. In contrast to pastes, wet solids do not flow easily through syringe-needle combinations suitable for intradermal and / or intramuscular injection and may be prone to fracture and / or disintegration due to applied shear forces (e.g., using oscillatory and / or rotational rheometers), whereas pastes are flowable and generally spread uniformly under similar shear conditions.

[0095] To prepare a paste, a non-solvent fluid (diluent) is added to the powder in an amount sufficient to produce at least a coating of fluid around the powder particles. While this is an ideal situation in which all direct powder-to-powder contact between individual powder particles is completely blocked, it should be noted that in practice, many finely divided powders are highly cohesive (including within the particle size ranges described herein) and, despite powder processing (e.g., milling, sieving, crushing, etc.) and / or application of high shear mixing techniques, it may not be possible to completely block all direct powder-to-powder contact. Additional fluid may then be added to the mixture to fill the interstices (i.e., void volume) between the powder particles, thereby allowing the particles to flow as a fluid when the yield stress of the paste is exceeded. Thus, a powder with a very low density (i.e., a high surface area to volume ratio) will require a larger amount of fluid to form a paste than a powder with a lower surface area to volume ratio. In the spectrum between suspensions and wet solids described above, pastes can be formed over a range of solids contents (e.g., 48-55%) within which the consistency of the paste may differ (e.g., increased firmness in the high solids content region compared to the low solids content region of the range), but the composition remains distinct from a suspension or wet solid (e.g., with respect to its flow characteristics and / or the resistance of the solid phase to settling over the relevant storage period). The solids content region / range in which pastes are observed / formed will vary depending on the properties of its constituent liquid and solid phases, with the solid phase generally having the greatest influence. As a non-limiting example, as described above, powders with lower density and / or larger specific surface area (e.g., as measured by nitrogen adsorption) may form pastes in the lower solids content range (e.g., 15-23%) than powders with higher density and / or smaller specific surface area that may form pastes in the higher solids content range (e.g., 58-64%). Despite the significant difference in solids content, both compositions may be distinguished from suspensions or wet solids, as discussed above.

[0096] Pastes are typically referred to as semi-solid and / or viscoelastic compositions, which are broad terms intended to describe the rheological properties / behavior of compositions / materials. These terms ("semi-solid" and / or "viscoelastic") can encompass a wide range of pharmaceutical dosage forms. Thus, although gels and pastes both have semi-solid properties and both may be referred to as semi-solids, they are physically different dosage forms. In particular, the solids concentration of pastes is typically much higher and the particles are generally much larger than the upper limit of the colloidal region (0.5 μm). Overall, the USP-NF defines at least six different dosage forms as semi-solid, including creams, foams, gels, jellies, ointments, and pastes. However, it will be readily recognized and understood by those skilled in the art that, despite their different physical compositions, these pharmaceutical dosage forms all have semi-solid properties and are therefore broadly referred to as semi-solids.

[0097] Pastes can be formed from the powders provided by the methods of the invention by mixing the powders with one or more fluid non-solvent materials. Non-solvent fluids suitable for use in preparing pastes according to this aspect of the invention include oils such as Miglyol 810 or 812, or other pharma- ceutically acceptable compounds, non-limiting examples of which include triacetin or benzyl benzoate. Preferred for use according to the methods of the invention are triacetin and / or Miglyol 812, which are mixed / blended with a powder containing one or more active pharmaceutical ingredients to produce a paste formulation, resulting in a paste having a solids concentration of at least about 30%, preferably at least about 40%, preferably at least about 50%, at least about 55%, at least about 60%, at least about 65%, or at least about 70%. Such pastes can then be loaded into commercially available syringes (either wide or narrow bore) fitted with commercially available needles of a gauge, wall thickness and length appropriate for the intended route of administration (e.g., 23G, 25G, 27G or 30G) and used to deliver relatively small amounts of the therapeutic paste via intradermal and / or intramuscular injection into animals.

[0098] As known in the art (and particularly as described elsewhere herein), it may be shown that the injection force and / or flow resistance for a given syringe, needle, and / or substance combination may be substantially dominated by viscous effects near the reservoir outlet, for example due to the abrupt change in cross-sectional area near this region as the needle lumen becomes much smaller as the syringe barrel becomes wider. Furthermore, it has been observed that pastes containing cohesive finely divided powders that tend to form robust aggregates (aggregates include two or more powder particles that are not fully dispersed during processing, mixing, and / or may form during long-term storage) may exhibit partial and / or complete clogging upon delivery of the paste from the syringe reservoir to the needle. Complete clogging completely prevents the flow of fluid from the device. In contrast, partial clogging does not completely prevent the flow of fluid, but may mean a sudden increase in force / pressure during delivery, which may result in stuttering during fluid delivery.

[0099] Thus, the present invention addresses these limitations by providing a method of producing both the starting material and the formulation that ultimately results in better flow of the paste from the syringe without resulting in complete clogging and particle agglomeration in the formulation.

[0100] In certain embodiments, the paste formulation is placed in a syringe reservoir, which may be made of any material suitable for the intended use and compatible with the paste formulation. Non-limiting examples of reservoir materials include glass (e.g., borosilicate glass) and plastic (e.g., polypropylene, polycarbonate, polystyrene, cyclic olefin polymers and copolymers, etc.). As described above, the reservoir may have any suitable dimensions, and any suitable volume of the reservoir may contain the paste. For example, in some embodiments, the paste is 15 μL to 1000 μL in volume. In some embodiments, the paste may be greater than 50 μL in volume, and in some embodiments, the paste may be greater than 100 μL in volume. In some embodiments, the paste may be greater than 1000 μL in volume, and in some embodiments, the paste may be greater than 2000 μL in volume. The volume of paste placed in the reservoir may sometimes be referred to as the injection volume (e.g., when substantially the entire amount of the paste is injected and / or ejected from the syringe).

[0101] In certain embodiments, a syringe with an integrated needle (i.e., attached to the syringe as a permanent fixture, not removably attached, such as via a luer lock) can be suitably used to deliver the formulation of the present invention. Such syringe / needle combinations appropriately take into account the above-mentioned syringe and needle size requirements. Syringe / needle combinations useful according to such aspects of the invention are commercially available, for example, from Becton Dickinson or Medtronic / Covidien.

[0102] Pastes suitable for use in accordance with the present invention may have any suitable material characteristics (e.g., solids concentration, solids content, viscosity profile, density, and / or the like). For example, in some embodiments, the pastes may have a solids concentration of greater than 100 mg / mL, greater than 200 mg / mL, or between 300 and 500 mg / mL (e.g., greater than any one of or between any two of 100, 125, 150, 175, 200, 225, 250, 275, 300, 325, 350, 375, 400, 425, 450, 475, 500, 525, 550, 575, 600, 625, 650, 675, 700, 725, 750, 775, 800, 825, 850, 900, 950, 1000 mg / mL or more). As a further example, the paste may have a solids content (e.g., weight of powder relative to the total weight of the paste) of about 30% to about 70% (e.g., greater than or between any two of 1, 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85% or more, preferably about 30%, about 35%, about 40%, about 45%, about 50%, about 55%, about 60%, about 65% or about 70%). As yet another example, the paste may have a density of 1.0 to 1.4 g / mL (e.g., 1.20 g / mL) (e.g., greater than or between any two of 1.0, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, 2 g / mL or more). With respect to the active pharmaceutical ingredient ("API") of the paste, the paste of the present invention may have an API content of at least about 20% by weight of the paste, e.g., from about 20% to about 70% API, from about 30% to about 65% API, from about 35% to about 60% API, from about 20% to about 50% API, from about 25% to about 50% API, from about 20% to about 40% API, from about 30% to about 40% API, or from about 30% to about 35% API (including all values ​​within these ranges).

[0103] In certain aspects, a suitable paste may be a protein paste with a solids content of 35%, a density of 1.15 g / mL, and a solids concentration of approximately 400 mg / mL. In certain other aspects, a suitable paste may be a protein paste with a solids content of 50%, a density of 1.25 g / mL, and a solids concentration of approximately 625 mg / mL. In certain other aspects, a suitable paste may be a protein paste with a solids content of 65%, a density of 1.3 g / mL, and a solids concentration of approximately 845 mg / mL. In one example, such pastes can be tested, characterized, or optimized by dispensing the paste from a variety of syringes with needles of different exposed lengths (e.g., 0.25, 0.5, 1.0 inches), such as 21, 22, 23, 25, or 27 gauge standard wall, thin wall, extra thin wall, special thin wall needles, etc., to determine the optimal commercially available needle and syringe combination for delivering small volumes of a particular paste while minimizing the injection force required to subcutaneously deliver the paste to an animal.

[0104] Volumetric flow rates (e.g., microliters per second (μL / s)) may vary depending on the cross-sectional dimensions of the reservoir and the speed of the plunger. Thus, volumetric flow rates between syringes with different reservoirs may be matched by varying the plunger speed applied between the syringes. For example, a syringe with a reservoir with a smaller internal lateral dimension (e.g., a 100 μL capacity reservoir) may require a higher plunger speed to achieve a given volumetric flow rate than a syringe with a reservoir with a larger internal lateral dimension (e.g., a 1000 μL capacity reservoir). As a further example, for four exemplary syringes with reservoirs of various capacities and internal dimensions, Table 1 provides the respective plunger speeds required to achieve two specific volumetric flow rates of 33.3 μL / s and 67.0 μL / s.

[0105] Table 1. Plunger speeds at two exemplary flow rates for reservoirs matching specific syringes TIFF2024534963000001.tif53128

[0106] As shown, in some embodiments, the syringe is configured to dispense paste at a flow rate of greater than 30 μL / s when the plunger moves at a speed between 2 and 40 mm / s, and, as shown in the illustrated example, the paste flow rate is substantially linearly proportional to the speed of plunger movement.

[0107] Some embodiments of the present methods for intradermally injecting a quantity of paste include displacing a plunger of a syringe to expel the paste from a reservoir of the syringe through a lumen of a needle of the syringe, the reservoir having an internal first lateral dimension greater than an internal second lateral dimension of the lumen, the second lateral dimension being between 0.1 and 0.9 mm, the paste having a solids concentration greater than 100 mg / L, and the paste expelled at a flow rate greater than 30 μL / s when the plunger moves at a speed between 2 and 40 mm / s. Some methods include removing a sealing cap from a fitting of the reservoir (e.g., a luer fitting). Some methods include coupling a needle to the reservoir via a luer fitting disposed on at least one of the needle and the reservoir. Some methods include placing the needle into and / or through the skin tissue of the patient.

[0108] In some methods, the paste injection volume is greater than 10 μL. In some methods, the paste injection volume is between 15, 500, or 1000 μL and 1200, 2000, or 3000 μL. In some methods, the paste injection volume is between 30 μL and 100 μL.

[0109] Pharmaceutically Active Ingredients The compositions of the present invention suitably contain one or more (e.g., 1, 2, 3, 4, 5 or more) pharma- ceutically active ingredients (used interchangeably herein as "active pharmaceutical ingredients" or "therapeutic ingredients"). By "pharma-ceutically active ingredients" is intended an ingredient in a composition that has a physiological, metabolic, physical, or mechanical effect when introduced into an animal (e.g., a human or veterinary animal), and is therefore useful in therapeutic and diagnostic methods for treating, ameliorating, preventing, and / or diagnosing a disease or disorder in the animal into which the pharma-ceutically active ingredient is introduced. Examples of suitable pharma-ceutically active ingredients for use in preparing the paste formulations provided by the present invention include, but are not limited to, peptide, protein, and small molecule therapeutic or diagnostic agents.

[0110] In certain embodiments, the pharma- ceutical active ingredient is a peptide or protein therapeutic. Exemplary peptide or protein therapeutics include those approved for use in human and / or veterinary animal therapeutic and / or diagnostic applications, such as therapeutic peptides and proteins listed in the online "THPdb" database (available at http: / / crdd.osdd.net / raghava / thpdb / ). Such peptide and protein therapeutic agents include enzymes (such as dornase alfa, velaglucerase alfa, taliglucerase alfa, asparaginase, glucarpidase, asfotase alfa, elosulfase alfa, sebelipase alfa, sacrolase, and pegloticase), antithrombin agents (such as lepirudin, bivalirudin, defibrotide, and sulodexide), thrombolytic agents (such as reteplase, anistreplase, tenecteplase, streptokinase, and urokinase), peptide or protein hormones (such as parathyroid hormone, amylin, angiotensin, growth hormone, growth hormone releasing factor, glatiramer, exenatide, insulin-like growth factor, cosinotropin, chorionic gonadotropin (e.g., human chorionic gonadotropin), and growth hormone), bone active peptides or proteins (calcitonin, e.g., salmon calcitonin, diabetogenic peptides or proteins, such as insulin (which may be human or porcine) and analogues thereof, including insulin lispro, insulin glargine, insulin aspart, insulin detemir, and insulin glulisine, pramlintide, and glucagon and analogues thereof, including dasiglucagon; antibodies or fragments thereof, which may be monoclonal antibodies or fragments thereof, such as cetuximab, thrombin, rastuzumab, bevacizumab, rituximab, obinutuzumab, gemtuzumab, canakinumab, ipilimumab, daratumumab, vedolizumab, ustekinumab, siltuximab, ramucirumab, pembrolizumab, ofatumumab, nivolumab, mepolizumab, brodalumab, pertuzumab, denosumab, golimumab, belimumab, raxibacumab, blinatuomab, dinutuximab, and ibritumomab), non-antibody antitumor agents (leuprolide,such as denileukin diftitox, aldesleukin, asparaginase, pegaspargase, interferon beta, afibercept, lenograstim, and sipuleucel-T), infertility drugs (such as leuprolide, menotropins, lutropin alfa, follitropin beta, urofollitropin, and chorionic gonadotropin alfa), and immunosuppressants (such as etanercept, peginterferon alfa, interferon alfa, filgrastim, pegfilgrastim, sargramostim, anakinra, interferon beta, interferon gamma, adalimumab, infliximab, basiliximab, muromonab, efalizumab, daclizumab, abatacept, rilonacept, belatacept, natalizumab, brinzumomab, ustekinumab, and human immunoglobulin). Other protein and peptide therapeutics suitable for use in the compositions and methods of the invention will be familiar to those of skill in the art. Protein and peptide therapeutics advantageously used in accordance with the present invention may be naturally derived, synthetically produced, or recombinantly produced using methods for producing peptides and proteins well known in the art.

[0111] Other active pharmaceutical ingredients suitable for use in preparing the compositions of the invention by the methods of the invention are small molecule therapeutic and / or diagnostic agents and their salts. Such agents are typically low molecular weight (e.g., less than about 1000 daltons) organic or inorganic compounds that, when introduced into an animal (e.g., a human or veterinary animal), have a desired biological activity that renders them useful in treating, ameliorating, preventing and / or diagnosing a disease or disorder. Examples of such small molecule active pharmaceutical ingredients (and their salts) suitable for use in the present invention include epinephrine, benzodiazepines, catecholamines, "triptans," sumatriptan, novantrone, chemotherapeutic small molecules (e.g., mitoxantrone), corticosteroid small molecules (e.g., methylprednisolone, beclomethasone dipropionate), immunosuppressive small molecules (e.g., azathioprine, cladribine, cyclophosphamide monohydrate, etc.). , methotrexate), anti-inflammatory small molecules (e.g., salicylic acid, acetylsalicylic acid, lisofylline, diflunisal, choline magnesium trisalicylate, salicylate, benorylate, flufenamic acid, mefenamic acid, meclofenamic acid, triflumic acid, diclofenac, fenclofenac, alclofenac, fentiazac, ketorolac, ibuprofen, flurbiprofen, ketoprofen, naproxen, fenoprofen, fenoprofen, ambufen, suprofen, indoprofen, tiaprofenic acid, benoxaprofen, pirprofen, tolmetin, zomepirac, clopinac, indomethacin, sulindac, phenylbutazone, oxyphenbutazone, azapropazone, feprazone, piroxicam, isoxicam), small molecules used to treat neurological disorders (e.g., cimetidine, ranitidine, famotidine, nizatidine, tacrine, metrifonate, rivastigmine, amine, selegiline, imipramine, fluoxetine, olanzapine, sertindole, risperidone, valproate semisodium, gabapentin, carbamazepine, topiramate, phenytoin), small molecules used to treat cancer (e.g., vincristine, vinblastine, paclitaxel, docetaxel, cisplatin, irinotecan, topotecan, gemcitabine, temozolomide, imatinib, bortezomib), statins (e.g.,atorvastatin, amlodipine, rosuvastatin, sitagliptin, simvastatin, fluvastatin, pitavastatin, lovastatin, pravastatin, simvastatin), and other taxane derivatives, small molecules used to treat tuberculosis (e.g., rifampicin), small molecule antifungals (e.g., fluconazole, ketoconazole), small molecule anxiolytics and small molecule anticonvulsants (e.g., lorazepam), small molecule anticholinergics (e.g., atropine, riboflavin), small molecule antimycotics (e.g., rifampicin), and other anticancer drugs (e.g., rifampicin). Small molecule therapeutic and diagnostic agents suitable for use in the compositions and methods of the present invention include, but are not limited to, small molecule beta agonists (e.g., albuterol sulfate), small molecule mast cell stabilizers and small molecule drugs used to treat allergies (e.g., cromolyn sodium), small molecule anesthetics and small molecule antiarrhythmics (e.g., lidocaine), small molecule antibiotics (e.g., tobramycin, ciprofloxacin), small molecule antimigraine drugs (e.g., sumatriptan), and small molecule antihistamines (e.g., diphenhydramine). Other small molecule therapeutic and diagnostic agents and salts thereof suitable for use in the compositions and methods of the present invention will be familiar to those of skill in the art. Additional formulations include combinations of such agents, including at least two of the small molecule therapeutic and diagnostic agents described herein and other agents familiar to those of skill in the art. Small molecules and their salts that can be advantageously used according to the present invention may be commercially obtained from a wide variety of sources (e.g., ThermoFisher, Aldrich Chemical, etc.) or may be synthesized using chemical and biochemical synthesis methods well known in the art.

[0112] How to use The compositions of the present invention can be used to treat, ameliorate, prevent or diagnose various diseases and disorders in animals, including veterinary animals or humans, in need of treatment, amelioration, prevention and diagnosis. A preferred such method involves administration of a relatively small amount (e.g., 1 μL to 10,000 μL or less) of one or more paste compositions of the present invention, preferably by intradermal, subcutaneous or intramuscular injection, resulting in potentially less and / or more rapid delivery of a bolus of therapeutic compound than can be achieved using other methods of administration of aqueous therapeutic formulations having lower concentrations of active ingredients, for example, via intravenous injection. These methods of use may reduce discomfort in animals after injection and may also exhibit certain pharmacokinetic and pharmacodynamic advantages, as detailed in the examples below. Diseases and disorders that are preferably treated, prevented, ameliorated or diagnosed using the paste compositions of the present invention will be readily apparent to those of skill in the art, and the selection of active ingredients to be used as starting materials in producing the pastes of the invention will also be familiar to those of skill in the art based on the disease, disorder or condition to be treated, prevented, ameliorated or diagnosed using the paste compositions of the present invention.

[0113] In some embodiments, the exemplary method of the present invention comprises administering to a subject having or at risk of hypoglycemia a paste formulation or composition as described herein in an amount effective to treat or prevent hypoglycemia. In some embodiments, the subject is administered a paste formulation comprising glucagon. In certain aspects, hypoglycemia may be caused by, or the patient may be at high risk of hypoglycemia due to, diabetes or non-diabetic related diseases, conditions, and disorders.

[0114] As described by the American Diabetes Association and Endocrine Society workgroup on hypoglycemia (Seaquist et al., Diabetes Care 36: 1384 - 1395 (2013)), a single plasma glucose concentration threshold that defines hypoglycemia in diabetes typically does not apply because hypoglycemic episode-specific glycemic thresholds shift to lower plasma glucose concentrations after recent preceding hypoglycemia (among other responses) and to higher plasma glucose concentrations in patients with poorly controlled diabetes and infrequent hypoglycemia.

[0115] However, warning values ​​can be defined that draw the attention of both patients and caregivers to the potential harms associated with hypoglycemia. Patients at risk for hypoglycemia (i.e., those treated with sulfonylureas, glinides, or insulin) should be aware that hypoglycemia may occur if self-measured plasma glucose concentrations or subcutaneous glucose concentrations from continuous glucose monitoring are ≦70 mg / dL (≦3.9 mmol / L). This is higher than the symptomatic glycemic threshold in both nondiabetic and well-controlled patients, and generally provides a time window to prevent clinical hypoglycemic episodes and some margin for the limited accuracy of monitoring devices at low glucose levels.

[0116] Severe hypoglycemic states are events that require the assistance of others to aggressively administer carbohydrates, glucagon, or take other corrective measures. Although there may be no plasma glucose concentration during the event, neurological recovery after plasma glucose returns to normal is considered sufficient evidence that the event was triggered by a low plasma glucose concentration. Typically, these events begin to occur at plasma glucose concentrations of ≤50 mg / dL (≤2.8 mmol / L). Documented symptomatic hypoglycemia is an event with typical symptoms of hypoglycemia accompanied by a measured plasma glucose concentration of ≤70 mg / dL (≤3.9 mmol / L). Asymptomatic hypoglycemia is an event without typical symptoms of hypoglycemia, but with a measured plasma glucose concentration of ≤70 mg / dL (≤3.9 mmol / L). Presumed symptomatic hypoglycemia is an event with typical symptoms of hypoglycemia, but without a plasma glucose measurement, possibly triggered by a plasma glucose concentration of ≤70 mg / dL (≤3.9 mmol / L). Pseudohypoglycemia is an event in which a diabetic patient reports any of the classic symptoms of hypoglycemia while the measured plasma glucose concentration is >70 mg / dL (>3.9 mmol / L) but approaching that level.

[0117] Indications that may be treated by the disclosed invention further include hypoglycemia-associated autonomic failure (HAAF). As described in Philip E. Cryer, Perspectives in Diabetes, Mechanisms of Hypoglycemia-Associated Autonomic Failure and Its Component Syndromes in Diabetes, Diabetes, Vol. 54, pp. 3592-3601 (2005), "recent antecedent iatrogenic hypoglycemia leads to both erroneous glucose counterregulation (by reducing epinephrine response to a given level of subsequent hypoglycemia in the absence of insulin reduction and glucagon increase) and hypoglycemia unawareness (by reducing sympathoadrenal and resulting neurogenic symptom response to a given level of subsequent hypoglycemia), thus resulting in a vicious cycle of hypoglycemia." HAAF affects type 1 diabetes and advanced type 2 diabetes patients. In addition, the disclosed invention may also treat hypoglycemia in patients after islet cell transplantation.

[0118] The compositions of the present invention can also be used to treat or prevent hyperinsulinemic hypoglycemia, which broadly refers to the state and effects of low blood glucose levels caused by excess insulin. The most common type of severe, but typically transient, hyperinsulinemic hypoglycemia occurs in type 1 diabetic patients with exogenous insulin administration. This type of hypoglycemia can be defined as iatrogenic hypoglycemia and is a limiting factor in glycemic control of type 1 and type 2 diabetes. Night-time hypo is a common type of iatrogenic hypoglycemia that occurs in patients taking exogenous insulin. However, hyperinsulinemic hypoglycemia can also be caused by endogenous insulin, for example, congenital hyperinsulinism, insulinoma (insulin-secreting tumor), exercise-induced hypoglycemia, and reactive hypoglycemia. Reactive hypoglycemia is a non-diabetic form of hypoglycemia that is caused by hypoglycemia that occurs after a meal, typically within 4 hours after a meal. Reactive hypoglycemia is also called postprandial hypoglycemia. Symptoms and signs of reactive hypoglycemia include hunger, weakness, tremors, drowsiness, sweating, confusion, and anxiety. Gastric surgery (e.g., bariatric surgery) is one possible cause, as food may reach the small intestine too quickly after surgery (e.g., post-bariatric hypoglycemia (PBH)). Other causes include enzyme deficiencies that make it difficult for the body to break down food, or increased sensitivity to the hormone epinephrine.

[0119] In some embodiments, the disease, condition, or disorder treated or prevented by the paste composition 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 another type of hypoglycemia as described herein and known to those skilled in the art. In some embodiments, the disease, condition, or disorder is diabetes.

[0120] In some embodiments, the method of the present invention comprises treating diabetes by administering to a subject having diabetes a therapeutic agent in a paste formulation described herein in an amount effective to treat diabetes. In some embodiments, the subject is administered a paste formulation comprising insulin. In some embodiments, the subject is administered a paste formulation comprising pramlintide. In some embodiments, the subject is administered a paste formulation comprising insulin and pramlintide. In some embodiments, the subject is administered a paste formulation comprising exenatide. In some embodiments, the subject is administered a paste formulation comprising glucagon and exenatide.

[0121] In certain aspects, the paste formulation of the present invention that contains epinephrine can be administered to subjects who are at risk of anaphylaxis or suspected of anaphylaxis.Epinephrine is indicated as an emergency treatment for type I allergic reaction, which can be caused by multiple causes, including but not limited to food, drug and / or other allergens, allergen immunotherapy, diagnostic test materials, insect stings and bites, and idiopathic or exercise-induced anaphylaxis.

[0122] Other diseases, disorders and conditions that are suitably treated, prevented, ameliorated or diagnosed using the compositions and methods of the present invention will be familiar to those of skill in the art and include, but are not limited to, cancer, infectious diseases, bacterial diseases, fungal diseases, viral diseases, and other diseases, disorders and conditions involving inflammatory, neurological, osteological, gastrointestinal, circulatory, cardiovascular, skin, muscular, developmental and other symptoms, signs or dysfunction.

[0123] The present invention has been described herein with reference to exemplary embodiments. Although certain embodiments have been described above with a certain degree of specificity or with reference to one or more individual embodiments, those skilled in the art will be able to make numerous modifications to the disclosed embodiments without departing from the scope of the present invention. Thus, the various exemplary embodiments of the method and system are not intended to be limited to the particular forms disclosed. Rather, they include all modifications and alternatives that fall within the scope of the claims, and embodiments other than those shown may include some or all of the features of the embodiments shown. It will be readily apparent to those skilled in the art that other suitable modifications and adaptations to the methods and applications described herein may be made without departing from the scope of the invention or any of its aspects. Similarly, it will be understood that the benefits and advantages described above may relate to one or several aspects. Although the present invention has been described in detail above, the present invention will be more clearly understood by reference to the following examples, which are included herein for illustrative purposes only and are not intended to be limiting of the present invention. EXAMPLES

[0124] The following examples and drawings are included to illustrate preferred embodiments of the invention. Those skilled in the art should recognize that the techniques disclosed in the examples or drawings represent techniques discovered by the inventor to function well in the practice of the invention, and thus can be considered to constitute preferred embodiments for its practice. However, those skilled in the art should recognize that in light of this disclosure, changes can be made to the specific embodiments disclosed herein without departing from the spirit and scope of the invention.

[0125] Example 1: Effect of manufacturing process on the stability of spray-dried powders containing proteins Previous work by some of the present inventors established initial processes for preparing injectable paste formulations containing relatively high concentrations of certain proteins and peptides (see, e.g., U.S. Patent Nos. 8,790,679, 8,110,209, and 9,314,424, and U.S. Patent Application Publication Nos. 2017 / 0007675 and 2017 / 0216529, all of the disclosures of which are incorporated herein by reference in their entireties). In this work, the effect of various manufacturing parameters on the preparation and stability of spray-dried powders and pastes containing high concentrations of antibodies was investigated with the goal of optimizing the manufacturing process for use in the preparation of storage-stable commercial formulations containing high concentrations of therapeutically active ingredients, particularly high molecular weight proteins such as antibodies.

[0126] Different paste formulations containing spray-dried powders containing IgG antibodies were first prepared by spray drying aqueous IgG solutions (e.g., aqueous "feed" solutions containing 20 mg / mL IgG) containing various carriers and excipients at different pH values ​​(measured before spray drying), details of which are shown in Table 2 below.

[0127] (Table 2) Representative powder compositions TIFF2024534963000002.tif59164

[0128] The two formulations were then spray dried using the following settings and under the conditions shown in Table 3.

[0129] Table 3: Spray drying conditions for two exemplary formulations TIFF2024534963000003.tif58164

[0130] Unless otherwise noted, all spray drying studies discussed in this application were performed using a BUCHI Corporation B-290 Mini Spray Dryer equipped with a standard two-fluid nozzle. This spray dryer contains a built-in floating ball flow meter for the nozzle (atomization) gas flow rate on a scale of 0-60 mm. Conversions to standard units of air flow rate (L / hr) and liquid flow rate (mL / min) are provided in the B-290 operating manual and the aspirator air flow rate and liquid feed pump settings are entered into the B-290 instrument on a scale of 0-100%.

[0131] After preparation of these two spray-dried powder formulations, samples were subjected to scanning electron microscopy (SEM) to examine the particle morphology of the two formulations. As seen in Figure 1, SEM micrographs of the two formulations showed clear differences in particle morphology. The particles of Formulation 1 exhibited an irregular shape with extensive pitting, whereas the particles of Formulation 2 exhibited a more regular spherical shape without apparent pitting. Additionally, on average, the particles of Formulation 1 were slightly larger than those of Formulation 2. Taken together, these physical characteristics indicate that the particles of Formulation 1 have a greater surface area than those of Formulation 2.

[0132] Pastes were then prepared from these two formulations by adding Miglyol 812N to them to the point where a paste composition was obtained but before the particles were converted into a suspension that could settle over time due to the relative excess of liquid to the solid phase. In formulation 1, the solids content range in which pastes were formed was significantly lower than the range in which pastes were formed in formulation 2. Formulation 1 formed pastes with solids contents ranging from approximately 38% to 42% w / w, exhibiting a solids concentration of 475 mg / mL solids and an IgG content of approximately 400 mg / mL. In contrast, in formulation 2, a paste containing a solids content of 65% w / w was prepared, exhibiting a solids concentration of approximately 820 mg / mL and an IgG concentration of approximately 630 mg / mL. These results suggest that formulation 1 produced a lower concentration of paste (in terms of both solids concentration and IgG concentration) than formulation 2 due to the physical characteristics of the spray-dried powder of formulation 1 (e.g., a larger surface area due to surface wrinkles).

[0133] To assess the effect of solids content on the injection force required to deliver the pastes through a hypodermic needle (modeling the injection of therapeutic paste formulations into animals), these pastes were loaded at 1 mL into a glass syringe (inner diameter approx. 4.6 mm) and delivered through a 27G ultra-thin walled ¼ inch (exposed length approx. 6 mm) needle secured to the syringe via a Luer lock fitting. The force required to deliver 1 mL of paste from the syringe in 30 seconds (e.g., volumetric flow rate of 33.3 μL / sec) was measured using a texture analyzer (force is plotted against distance traveled by the plunger). Formulation 1 (42% w / w solids, approx. 400 mg / mL IgG) required an injection force of approximately 36 N to expel 1 mL of paste in 30 seconds, whereas formulation 2 (65% w / w solids, 630 mg / mL IgG) required an injection force of nearly 60 N to expel 1 mL of paste in 30 seconds, partially reflecting their difference in overall solids content.

[0134] Lower injection forces may facilitate delivery and improve overall ease of administration. Therefore, it would be desirable to produce paste formulations with high active ingredient (e.g., mAb) concentrations that can be delivered with relatively low injection forces using commercially available syringe / needle combinations. To evaluate the impact of other spray drying process parameters on the ability to produce stable, syringeable therapeutic protein paste formulations, the inventors evaluated the levels of protein aggregation in nonspecific IgG formulations before and after spray drying of initial solutions at different pH. A 20.0 mg / mL IgG aqueous feed solution was prepared in the "Formulation 2" solution described above in Table 3, either at pH 4 (using citrate buffer) or pH 6 (using histidine buffer). Size exclusion chromatography was then used to determine the relative percentage of mAb aggregates after spray drying. In both formulations, the aggregation levels after spray drying were approximately 2.6% for the pH 6 formulation and 1.9% for the pH 4 formulation, indicating that the pH of the starting feed solution can drive a measurable difference in the % protein aggregation rate of the final spray-dried powder. For certain IgG formulations, the preferred pH to minimize aggregate formation may range from 3.5 to 4.0.

[0135] To further evaluate the effect of the spray drying process parameters themselves, a series of experiments were performed to investigate the effect of inlet temperature, aspirator flow rate, and nozzle pressure gas flow rate during the spray drying process of IgG solutions on the resulting morphology (sphericity), size, and size distribution of IgG particles in the resulting spray-dried powder. Eight sets of process parameters were evaluated using a 20 mg / mL nonspecific IgG starting solution in the above formulation 2 solution at pH 6.0 (Table 4). All samples were filtered prior to spray drying and then subjected to secondary drying under vacuum.

[0136] Table 4. Spray drying process parameters TIFF2024534963000004.tif59161

[0137] After spray drying, the appearance of the resulting powder was visually assessed and then solubilized (dissolved) in water for injection (WFI) at 1 mg / mL and assessed for dissolution time and percent aggregation via size exclusion chromatography (SEC). The results are shown in Table 5.

[0138] Table 5. Effect of spray drying parameters on the appearance of IgG powder TIFF2024534963000005.tif93134

[0139] As can be seen in Table 5, the powders prepared from the different feed solutions evaluated in this study contained similar percentages of protein aggregates measured upon dissolution, and each powder also showed similar dissolution times within a narrow range of 22–27 seconds.

[0140] To examine these powders more closely at the particle level, samples of powder from each of the eight formulations listed in Table 5 were examined using Scanning Electron Microscopy (SEM) to assess particle size, particle shape, and the distribution of each within a given sample. The results are shown in Figure 2.

[0141] As seen in Figures 2A-2D, formulations 5-8, all with a spray dryer inlet temperature of 90°C, showed a range of particle sizes and morphologies, with a mixture of small and large particles that exhibited a mixture of spheres and donut shapes. In contrast, as seen in Figures 2E-2H, formulations 5-8, all with a spray dryer inlet temperature of 70°C, showed more uniform particles in terms of size and shape, with most particles appearing spherical, with only a few donut-shaped particles observed. Of these formulations, formulations 7 and 8 appeared to be the most uniform in size, exhibiting a majority of small spherical particles.

[0142] These results were confirmed when the particle size distribution was measured by individually measuring the diameter (on a representative SEM micrograph) of approximately 200 IgG particles per formulation, and the results of these measurements are shown in Figures 3-4. As can be seen in Figure 3, formulations 1-4, which had a spray dryer inlet temperature of 90 °C, exhibited a larger particle size distribution (Figures 3A-3D) than formulations 5-8, which had a spray dryer inlet temperature of 70 °C (Figures 3E-3H). Interestingly, for a given spray dryer inlet temperature, higher gas flow rates tend to produce smaller and more uniform particles; for example, compare formulations dried at 40 mm gas flow (Figures 3A, 3C, 3E, and 3F) with their corresponding formulations dried at 60 mm gas flow (Figures 3B, 3D, 3G, and 3H, respectively).

[0143] Taken together, the results of these studies indicate that spray drying settings can affect the production of spherical particles with an appropriate size distribution. In particular, inlet temperatures of about 70-90°C, more specifically about 70-80°C, and gas flow rates of about 40-60 mm, especially about 60 mm, appeared to produce smaller and more spherical particles in the powder. Indeed, an inlet temperature of about 70°C was determined to be the lowest temperature that could be used to obtain spherical particles in the B-290 spray dryer; however, using lower temperatures with the feed and process parameters described in this example, which would have been expected to make the protein or peptide less susceptible to temperature denaturation, actually had the effect of producing more donut-shaped particles with a non-uniform size distribution (data not shown). The ability to produce smaller spherical particles with an appropriate size distribution is important for the ultimate production of high solids content, high protein concentration pastes that can be suitably injected in relatively small amounts into animals, particularly humans, for therapeutic and diagnostic purposes.

[0144] Example 2: Effect of formulation excipients on the manufacture and injectability of spray-dried proteins Beyond the spray dryer settings, our results showed that the components of the formulation being spray dried can affect the physical and performance properties of the spray-dried powder prepared from a given formulation. To further explore and optimize the impact of these formulation components, a representative monoclonal antibody (Trastuzumab or "TmAb", the API in the marketed drug Herceptin®) was formulated in the presence of various excipients and buffers, pharma- ceutically acceptable carriers or bulking agents, surfactants, etc., and the impact of each excipient on the degree of formulation aggregation both at time 0 after powder production by spray drying and upon powder storage was evaluated. TmAb is a protein with a size of 148 kDa in monomeric form, but when aggregated, it forms larger molecular weight dimers and other multimers that are not only immunogenic themselves, but also act as nucleation centers for the formation of even larger aggregates in solution; such aggregates may be immunogenic and / or may be removed by the immune system before the antibody has a chance to exert a therapeutic effect. From a therapeutic perspective, it would therefore be ideal to be able to prepare (e.g. by spray drying) a powder containing TmAb that has a low degree of aggregation on storage, which could be suitable for use in high solids content / high concentration pastes for injection into animals, including humans.

[0145] For preparation, the commercially available TmAb solution was dialyzed overnight at 4° C. against the desired formulation buffer (see table below) with constant stirring (50 kDa molecular weight cut-off). The dialyzed TmAb solution was then spray dried at an inlet temperature of 70° C., nozzle flow rate of 40 mm, aspirator setting of 85%, and feed pump at 10% (approximately 3 mL / min). Once the powder was produced, the powder was secondary dried (e.g., under vacuum) at 150 mT, 5° C. for 1 day, then at 30° C. for 3 days to reduce the water content to a target level of <1% (w / w), and then the powder was stored in a glass vial that was backfilled with nitrogen and stoppered to create a closed system. Samples were stored at 40° C. for 4 days or at 50° C. for 4 hours and compared to the t=0 sample (immediately after lyophilization) for the extent of aggregation and other stability parameters by dissolving the powder sample in water to a concentration of 1 mg / mL TmAb and then analyzing the solution by size exclusion, ion exchange, and reverse phase chromatography.

[0146] In the first experiment, five formulations were prepared, each containing 20 mg / mL TmAb, 5.15 mg / mL trehalose, 0.05 mg / mL polysorbate 20, 0.412 mg / mL methionine, and the remainder excipients, with the pH adjusted to the values ​​shown in Table 6.

[0147] Table 6. pH and buffer type formulation (first time) TIFF2024534963000006.tif105170

[0148] These formulations were then evaluated by size exclusion chromatography for the extent of protein aggregation in the pre-spray dried formulation and in the solubilized spray dried powder. The results are shown in Table 7.

[0149] Table 7. Effect of pH and buffer type on TmAb aggregation (1st run) TIFF2024534963000007.tif36170

[0150] To expand on these studies, a second formulation was prepared containing the same levels of TmAb, trehalose and polysorbate 80 as described for the first one above, but containing either succinate or lactate buffer and with a pH between 4 and 6, as shown in Table 8.

[0151] Table 8. pH and buffer type formulation (2nd run) TIFF2024534963000008.tif104169

[0152] These formulations were then evaluated for the extent of protein aggregation in the pre-spray dried formulation and in the solubilized spray dried powder via size exclusion chromatography, comparing samples tested before spray drying with one taken at t=0 after spray drying and another stored at 50° C. for 1 day. The results are shown in FIG. 4. Combined with those from the first formulation, these results indicate that the optimal buffer and pH conditions for the pre-spray dried solution to minimize the amount of aggregation in both the pre-spray solution and the powder after spraying and drying are the use of a lactate buffer and a pH of about 3.5 to about 4.5, e.g., about 4.0, particularly for IgG-containing formulations.

[0153] Next, the effect of the amount of trehalose in the formulation before spray drying on aggregation in solution and in the powder after spray drying was evaluated. TmAb formulations (20 mg / mL) in 5 mM lactate buffer at pH 4.0 or 6.0 were prepared containing the amounts of trehalose and other excipients shown in Table 9.

[0154] Table 9. Formulations with different sugar contents TIFF2024534963000009.tif217160

[0155] These formulations were then evaluated for the extent of protein aggregation in the aqueous formulation (feed) before spray drying and in the powder after spray drying via size exclusion chromatography, comparing samples tested before spray drying with those taken at t=0 after spray drying and freeze drying and others stored at 50°C for 1 day. The results are shown in Figure 5. These results indicate that trehalose has a stabilizing effect on spray drying and storage after spray drying in that it reduces powder aggregation. However, a relatively high amount of trehalose is required in the formulation to achieve such stability, which reduces the active ingredient (in this case TmAb) in the paste formulation. Thus, the amount of trehalose included must be optimized along with other excipients and formulation components to increase stability while allowing for a relatively high active agent content in the paste formulation. The inventors also have results for formulations prepared under the conditions of this example that suggest that the use of sucrose in place of trehalose at similar concentrations may have a greater stabilizing effect on the powder formulation. Other excipients that may be advantageously used in a similar manner include amino acids, advantageously one or more naturally occurring amino acids, e.g., hydrophobic amino acids (which may help prevent the hydrophobic core of one TmAb molecule from binding to a second TmAb, thus reducing aggregation), acidic / basic amino acids such as arginine, which have been reported to have the ability to stabilize protein and peptide formulations even in the dry state, and sugar combinations such as dextran / trehalose co-formulations.

[0156] These studies continued to evaluate the effect of polysorbate 20 content in the formulation prior to spray drying on aggregation in the feed solution and in the powder following spray drying. In the first of these studies, TmAb formulations (20 mg / mL) were prepared in 4.8 mM histidine buffer at pH 6.0 with the polysorbate 20 and other excipient contents shown in Table 10.

[0157] Table 10: Evaluation of polysorbate 20 content TIFF2024534963000010.tif104170

[0158] Other formulations containing either polysorbate 20 or polysorbate 80 were also prepared as shown in Table 11.

[0159] Table 11: Polysorbate formulations (continued) TIFF2024534963000011.tif83153

[0160] These formulations were then evaluated for the extent of protein aggregation in the aqueous formulations prior to spray drying and in the solubilized spray-dried powders via size exclusion chromatography, comparing samples tested prior to spray drying with those taken at time 0 (e.g., within 1 day) after spray drying and freeze drying and others stored at 50°C for 1 day. The results are shown in Figures 6-7. These results indicate that polysorbate has a stabilizing effect on spray drying and post-spray drying storage stability (i.e., reducing powder aggregation). Under the conditions evaluated in this study, no significant difference was seen between polysorbate 20 and polysorbate 80 in stabilizing the powder. Furthermore, increasing the amount of polysorbate added to the formulation prior to spray drying only resulted in a slight increase in stability; approximately 0.5 mg / mL polysorbate 20 appears to provide a suitable improvement in stabilization without increasing the solids mass too much to the formulation (which would adversely affect (dilute) the amount of active agent, in this case TmAb, that may be included in the paste formulation, as described above for trehalose).

[0161] Next, the effect of including various amino acids as excipients on aggregation and storage stability was evaluated. First, various methionine-containing formulations were prepared according to the component amounts shown in Table 12.

[0162] Table 12. Methionine-containing formulations TIFF2024534963000012.tif108170

[0163] Using a similar procedure, the formulations prepared as shown in Tables 13 and 14 were evaluated when proline or glycine was included.

[0164] Table 13. Proline / glycine-containing formulations TIFF2024534963000013.tif113169

[0165] Table 14. Proline-containing formulations (continued) TIFF2024534963000014.tif80170

[0166] These formulations were then evaluated for the degree of aggregation in the formulations prior to spray drying and in the solubilized spray dried powder via size exclusion chromatography, comparing samples tested prior to spray drying with samples after spray drying and freeze drying (time 0 (T0) samples) and others stored at 50° C. for 1 day. The results are shown in FIG. 8 (for methionine), FIG. 9 (for proline / glycine 1st run), and FIG. 10 (for proline 2nd run). Taken together, these results indicate that while methionine (FIG. 8), proline (FIGS. 9 and 10), and glycine (FIG. 9) all act as stabilizing excipients under the conditions evaluated in this study, relatively large amounts of these amino acids may be required to see more than a minor effect on stability. Because increasing the content of these excipients in the feed solution would dilute the active ingredient in the powder and resulting paste formulations (while holding the mAb content relatively constant), these amino acids may not be the preferred excipients for these formulations manufactured and evaluated under the conditions of this example, as the trade-off of reducing the active agent content in these formulations in exchange for only a slight increase in stability is not worth it.

[0167] Finally, formulations containing cysteine ​​at various concentrations according to Tables 15, 16, and 17 were prepared.

[0168] Table 15. Cysteine-containing formulations TIFF2024534963000015.tif96163

[0169] Table 16. Cysteine-containing formulations (continued) TIFF2024534963000016.tif113163

[0170] Table 17. Cysteine-containing formulations (continued) TIFF2024534963000017.tif91163

[0171] These formulations were then evaluated for mAb aggregation in the pre-spray-dried formulation and in the solubilized spray-dried powder via size exclusion chromatography, comparing samples tested before spray drying with those taken at t=0 after spray drying and lyophilization and others stored at 50° C. for 1 day. The results are shown in FIGS. 11-12 and suggest that the inclusion of even small amounts of cysteine ​​in the pre-spray-dried formulation can have a significant positive effect on the storage stability of spray-dried powders prepared under the conditions described in this example.

[0172] To further evaluate the effect of cysteine, powders prepared from various cysteine-containing formulations were dissolved and then analyzed by ion exchange chromatography. These results are shown in Figures 13 and 14; Figure 13 shows the levels (as a percentage of the total as shown by AUC measurements) of the main peak (Figure 13A), acidic variants (Figure 13B) and basic variants (Figure 13C) present in cysteine-containing formulations of TmAb, while Figure 14 provides representative traces of the stability profile of two formulations, one containing 1.5 mg / mL cysteine ​​(Figure 14A) and the other containing 6 mg / mL cysteine ​​(Figure 14B). Taken together, these results indicate that high levels of cysteine ​​can disrupt the internal Cys-Cys bonds present within the TmAb molecule, which, in addition to loss of storage stability, can lead to inactivation of the antibody and, therefore, loss of its therapeutic efficacy. Therefore, low cysteine ​​content (e.g., 1.5 mg / mL) in the formulation before spray drying may improve stability while avoiding the loss of storage stability and potential loss of biological activity caused by the inclusion of high amounts of cysteine ​​in the formulation.

[0173] Finally, a dialysis solution of TmAb in the optimized excipient formulation was prepared and used to optimize certain spray drying parameters, specifically the optimal feed concentration and inlet temperature settings to maximize protein concentration and storage stability while minimizing aggregation. The formulations were prepared as shown in Table 18.

[0174] Table 18. Formulations for optimizing spray drying parameters TIFF2024534963000018.tif185160

[0175] Additional formulations were evaluated by ion exchange chromatography for aggregation both before spray drying and after spray drying at t=0 and t=1 day at 50° C. The results are shown in FIG. 15. These results indicate that using a higher feed concentration (e.g., 30 mg / mL TmAb vs. 20 mg / mL TmAb) produces a less aggregated starting solution, a less aggregated spray dried powder at a given spray dryer inlet temperature, and a more storage stable powder (e.g., compare the results in FIG. 15 for batch 52a vs. batch 29a; batch 52b vs. batch 29b; and batch 52c vs. batch 29c). Furthermore, for a given feed concentration, an inlet temperature of 70° C. appears to produce powders that are less agglomerated (t=0) and more storage stable (t=1 day at 50° C.) than those prepared at higher inlet temperatures (e.g., compare the results in FIG. 15 for batch 29a vs. batches 29b and 29c; and batch 52a vs. batches 52b and 52c); this result confirms the results reported above in Example 1 regarding the optimal inlet temperature.

[0176] Based on the studies described above, the composition detailed in Table 19 represents an example of a formulation that also provides high antibody drug concentrations (>400 mg / mL) while exhibiting good antibody stability in the resulting paste formulation.

[0177] Table 19. Xerreject™ Paste Formulation for High Concentration TmAb Paste TIFF2024534963000019.tif82159

[0178] Taken together, these studies provide examples of representative aqueous feed formulations and spray drying and process conditions for producing high-concentration, high-solids content, and storage-stable dry powder formulations of therapeutic proteins, such as monoclonal antibodies, suitable for use in preparing high-concentration injectable paste formulations that allow intradermal, subcutaneous, and / or intramuscular injection of therapeutic peptides and proteins that previously could only be administered intravenously over much longer periods. Indeed, using the approach described in this example, it is possible to prepare a matrix of formulations and equipment parameters that facilitate the screening of excipients and process parameters to facilitate the development of formulations suitable for the preparation of high-solids concentration formulations that can be advantageously spray dried into flowable, storage-stable paste formulations for therapeutic applications. This approach to preparing high-solids concentration injectable paste-like therapeutic formulations, a technology developed by Xeris Pharmaceuticals, Inc. based on its Xeriject™ technology platform, described herein, therefore provides many benefits to patients, including ease of administration, avoidance of discomfort, and potentially efficacy of therapeutic peptide / protein formulations.

[0179] Example 3: Preparation of a high-concentration paste containing therapeutic monoclonal antibodies Xeriject™ (XJ) is a proprietary formulation technology that can significantly increase the concentration and / or thermal stability of the active pharmaceutical ingredient (API) in a dose. With Xeriject™ technology, dry particles of the active pharmaceutical ingredient (API), preferably prepared by spray drying according to the methods described in Examples 1 and 2 above, are blended and mixed with a non-solvent liquid to form a paste. As previously described, pastes are biphasic compositions that exist in the spectrum between suspensions and wet solids, and solid concentrations in the powder can achieve drug concentrations of 30% w / w or more (250 mg / mL or more) with this approach. This technology represents a significant improvement over current therapies that often must be administered in the clinic as long-term IVs of low-concentration solutions. This technology can be used to deliver proteins, such as antibodies, or small molecules, in high doses to patients in a bolus subcutaneous administration. Furthermore, in certain formulations, Xeriject™ technology can result in improved thermal stability of the formulation (particularly the active pharmaceutical ingredient in the formulation) even at standard lower dose concentrations.

[0180] These studies evaluated Xeriject™ technology as a platform for the subcutaneous delivery of small amounts of therapeutic agents that previously could only be administered intravenously. Specifically, several commercially available monoclonal antibody products were formulated into a Xeriject™ paste formulation.

[0181] Table 20. Commercially available pharmaceutical products prepared into paste form using Xerreject™ technology TIFF2024534963000020.tif801701)5mg / kg, 70kg patient 2)3mg / kg, 70kg patient 3)3.6mg / kg, 70kg patient 4)3mg / kg, 70kg patient

[0182] To set a baseline, samples of three other commercial products, Herceptin® (TmAb; see Examples 1-2), Erbitux® (cetuximab; Eli Lilly and Company), and Pirivgen® (immunoglobulin; CSL Behring AG), were spray dried from the commercial formulations with minimal modifications and then examined under a scanning electron microscope to observe the morphology and size of the antibody particles in the powder products. Representative micrographs are shown in FIG. 16, which show particles observed in spray dried formulations of TmAb (FIG. 16A), cetuximab (FIG. 16B), and immunoglobulin (FIG. 16C), which are similar to those observed for the other therapeutic proteins in Example 1. Most of these commercial products exhibit a wide range of particle sizes, with the TmAb commercial formulation also exhibiting highly donut-shaped particles, which, based on the results shown in Example 1, would result in a powder material that is not optimal for use in preparing therapeutic pastes using the Xerivect™ formulation technology. Indeed, when the particle size distributions of these three formulations were obtained by laser diffraction, all formulations had polydisperse properties characterized by a range of particle sizes observed from the 10th to the 90th percentile, as shown in Figure 17.

[0183] To prepare Xerreject™ paste formulations of these commercial products, small batches of each were spray dried to prepare pastes according to the methods described above in Examples 1 and 2. Sucrose was added to the commercial cetuximab and immunoglobulin formulations instead of trehalose, and the cetuximab salt content was reduced by dialysis. The TmAb formulations were spray dried from the commercial formulations. Pastes were prepared from the spray-dried powders and triacetin (density = 1.16 g / mL) was included to give a final estimated paste density of approximately 1.24 g / mL. The total solids and active ingredient content in each paste formulation were then calculated to the values ​​shown in Table 21.

[0184] Table 21. Solids and active ingredient content of Xeriject™ mAb paste TIFF2024534963000021.tif26162

[0185] After preparation of the pastes, the formulations were evaluated for the injection force required to deliver 1 mL of a given paste in two different syringe / needle combinations: a 1 mL syringe with a stacked 23 gauge ½ inch standard wall (RW) needle (Gerresheimer AG; Bunde, Germany) and a syriQ BioPure® 1 mL long syringe with a 27 gauge ½ inch standard wall (RW) needle (Schott AG; Mainz, Germany). For each configuration, an injection force profile was generated using a plunger speed of 3.0 mm / sec (corresponding to a volumetric flow rate of approximately 98 μL / sec). Representative results for a preparation of Pirivigen® (immunoglobulin) prepared at 42% solids content with triacetin as diluent (continuous phase) are shown in FIG. 18. As expected, the injection force required to deliver 1 mL of paste with the larger (lower gauge) needle was significantly lower, confirming previous results with lower concentration paste formulations. Importantly, however, the entire 1 mL of paste product was delivered without the need for excessively large injection forces, even from a 27G needle. The ability to inject small volumes of therapeutic antibody paste formulations intradermally with relatively small gauge needles suitable for intradermal and / or intramuscular injection and relatively low injection forces significantly enhances the patient benefit of such injections in terms of reduced discomfort and the reduced time required to receive antibody therapy treatment, as compared to standard IV administration of such antibodies.

[0186] Next, the ability to formulate the TmAb into a usable, highly concentrated paste was evaluated. The commercial product was reconstituted with water and samples were either kept in solution and spray dried to a powder, or formulated into a paste (45.2% solids in triacetin) from the spray dried powder using the Xeriject™ technology described above. Evaluation of these samples by size exclusion chromatography demonstrated little difference between them (Figure 19) - all samples had both a main peak and fragments with identical retention profiles, while the Xeriject™ paste sample contained an additional peak corresponding to triacetin in the paste formulation. Thus, the spray drying and paste formation process did not result in aggregation of the TmAb protein when compared to the commercial product reconstituted in solution.

[0187] To evaluate the pharmacokinetics of these various TmAb formulations, particularly that of the Xeriject™ paste formulation, a commercially available antibody solution was spray dried as described in the previous examples to produce a spray-dried powder that was used to prepare a high solids paste as described above. The API loading in this formulation was 259 mg / mL and was administered by subcutaneous injection to male Sprague-Dawley rats at two different doses using a commercially available syringe / needle combination. A commercially available formulation of trastuzumab was also administered IV to control animals for comparison. As seen in Figure 20, IV administration of trastuzumab showed an initial spike in drug levels, while the Xeriject™ formulation had no spike and rose to steady drug levels after about 24 hours (Figure 20A). Over the next 6 days of sampling, the Xeriject™ formulation and IV administration maintained steady drug levels. As shown in Figure 20B, the pharmacokinetics of the Xeriject™ formulation was dose-dependent, with steady-state drug levels of 20 mg / kg Xeriject™ similar to the pharmacokinetics of 10 mg / kg IV administration. Cmax was slowed by approximately 15% for both the 10 mg / kg and 20 mg / kg doses of Xeriject™ TmAb compared to IV TmAb, while bioavailability (AUCo-t) of the 10 mg / kg and 20 mg / kg doses of Xeriject™ TmAb compared to 10 mg / kg IV TmAb was 39 and 45%, respectively (dose normalized). Finally, the T1 / 2 of the IV TmAb was approximately 10 days, while the T1 / 2 of the Xeriject™ formulation was longer than the time period allotted for the study and was therefore not determined here. As will be readily appreciated by those skilled in the pharmaceutical and medical arts, a blunted Cmax and sustained exposure (i.e., longer T1 / 2) may be favorable for compounds with a toxicity profile by Cmax and efficacy by AUC, providing another advantage of pastes prepared using the Xerreject™ technology of the present invention. These results demonstrate that bolus injections can administer large amounts of drug in relatively small volumes while achieving therapeutically beneficial circulating drug levels with kinetics similar to those achieved by IV administration (albeit for longer durations).The Xeriject™ approach thus significantly improves this particular drug therapy / delivery and may be similarly useful for other therapies that use large molecular weight peptides and proteins, such as therapeutic antibodies and enzymes.

[0188] Example 4: Preparation of a high solids paste containing a therapeutic enzyme To further explore the utility of the Xerreject™ technology provided by the present invention, pastes with high solids concentrations containing therapeutic enzymes were prepared. In these exemplary studies, multimeric PEGylated therapeutic enzymes were used as the active ingredient. In the first step, the PEGylated enzyme solution was converted to dry powder by freeze-drying and by the spray-drying process described in the previous examples. Samples of each powder preparation were then examined for morphology and size distribution by scanning electron microscopy. Representative micrographs are shown in Figure 21. As seen in Figure 21A, the PEGylated enzyme powder prepared by freeze-drying the feed solution exhibited large, irregular particles with a relatively high specific surface area. In contrast, as seen in Figure 21B, the powder prepared by the spray-drying process described herein exhibited small, spherical particles with a relatively low specific surface area. Furthermore, when pastes were prepared from both of these powders, the paste prepared from the spray-dried powder exhibited a higher solids concentration, and thus a higher enzyme concentration, than the paste prepared from the freeze-dried powder, as shown in Table 22.

[0189] Table 22. Enzyme paste properties TIFF2024534963000022.tif40168

[0190] These results are consistent with the previous examples herein, which show that powders containing more regular spherical particles with smaller size and surface area distributions are more suitable for the production of flowable pastes of active ingredients and ultimately for therapeutic subcutaneous injection into animals. Therefore, the remainder of these studies were carried out using spray-dried powders as the starting material for the production of therapeutic Xerreject™ pastes.

[0191] To study the pharmacokinetic parameters of the Xeriject™ enzyme paste formulation, the spray-dried paste enzyme preparation described above was administered by subcutaneous injection to male Sprague-Dawley rats using a commercially available syringe / needle combination. For comparison, an aqueous formulation of the enzyme (in PBS) was also administered IV and subcutaneously to control animals. Plasma enzyme concentrations in each group of animals were then determined over time, and the results are shown in Figure 22. The intravenously administered aqueous (PBS) formulation showed an initial spike of enzyme in plasma, which then rapidly declined over the next 48-72 hours (Figure 22A). In contrast, the subcutaneous aqueous (PBS) formulation and the Xeriject™ paste formulation showed similar pharmacokinetic profiles of a gradual rise to peak plasma concentration, followed by a longer and more gradual decline (Figure 22B). As shown in Table 23, the T1 / 2, Tmax, Cmax and AUC(0-t) values ​​for the subcutaneous formulations were also similar but significantly different from those of the IV administered enzyme, reminiscent of the results obtained with the IV aqueous vs. subcutaneous Xeriject™ paste formulations of TmAb described in Example 2 herein.

[0192] Table 23. Pharmacokinetic profiles of IV and subcutaneous enzyme formulations TIFF2024534963000023.tif52159

[0193] Pharmacodynamic analysis of plasma samples from treated animals showed differences not only between the IV (FIG. 23A) and subcutaneous (FIG. 23B) enzyme formulations, but also between the aqueous (PBS) and Xeriject™ paste formulations administered subcutaneously (FIG. 23B). Specifically, the aqueous and paste formulations exhibited similar pharmacokinetic profiles as described above, but the Xeriject® paste formulation exhibited a greater peak target reduction in the pharmacodynamic study (6 μM vs. 2 μM). Furthermore, results of the post-sacrifice pathology report (not shown) indicated that no significant injection site reactions (and thus post-injection discomfort) were observed in animals subcutaneously injected with the Xeriject™ paste formulation.

[0194] Taken together, these results demonstrate that paste formulations of highly concentrated, high molecular weight protein therapeutics provided by the present invention are useful in delivering controlled or sustained release depots of small amounts of therapeutic proteins subcutaneously in a manner that improves the patient experience beyond that obtained with conventional intravenous administration of such therapeutic proteins.

[0195] Example 5: Preparation of a high solids paste containing high concentrations of glucagon In further studies, a Xeriject™ paste formulation containing high concentrations of glucagon was prepared by thin film freezing (a particle engineering technique that produces a powder with a relatively high surface area) from an aqueous solution containing glucagon, trehalose, and a buffering agent (glycine) and the pH was adjusted to 3.0. The thin film lyophilized powder was formulated into a paste by gently grinding and mixing with sufficient triacetin to create a paste. 1 mg of glucagon was administered subcutaneously to rats in 5 μl of paste at the same dose as a larger volume (1 ml) of a commercially available aqueous glucagon formulation (Glucagon Emergency Kit or "GEK"; Eli Lilly). This allowed for a direct comparison of the pharmacokinetic and pharmacodynamic properties of the Xeriject™ paste formulation to the commercially available aqueous solution. The results are shown in FIG. 24.

[0196] Examination of the pharmacokinetic profiles of the two formulations (Figure 24A) showed that the Xeriject™ (Xeris) paste formulation, despite being injected in 200x smaller volumes, exhibited comparable pharmacokinetics to the aqueous formulation of glucagon injected in larger volumes. The same was true for the pharmacodynamic profile (Figure 24B), where the Xeriject™ (Xeris) paste formulation exhibited similar pharmacodynamic properties to those observed with the aqueous GEK formulation, despite being more slowly disintegrated over time as seen with other therapeutic peptides (see previous examples). These results indicate that the higher concentration peptide (glucagon) paste formulations provided by the present invention are useful in delivering small amounts of glucagon subcutaneously in a manner that improves the patient experience over that obtained with conventional intramuscular administration of aqueous formulations of glucagon, which requires injection of relatively large volumes.

[0197] Example 6: Preparation of a high solids paste containing insulin In addition to enabling injectable formulations with very high concentrations of therapeutic agents, the injectable pastes described herein may be utilized to significantly enhance the thermal stability of therapeutic agents, including those administered at relatively lower concentrations. One such example is human insulin, which is commercially available in concentrations ranging from u100 (approximately 3.5 mg / mL) to u500 (approximately 17.4 mg / mL). Insulin dosages vary from patient to patient, but generally range from 30 to 100 μL for u100 formulations. Thus, this therapeutic protein does not require a significant increase in drug concentration to reduce dosage. However, commercially available insulin pharmaceuticals are formulated as aqueous solutions that require refrigerated (2-8°C) transport and long-term storage conditions. This cold chain requirement may limit the availability of commercially available insulin in third world countries and compromise its quality. There is therefore a need for injectable insulin formulations with significantly improved thermal stability that can exhibit long-term stability at temperatures of at least 25, 30, or 35° C. (i.e., storage stability for at least one year, more preferably at least 18, 24, 30, or 36 months).

[0198] An example of the improved thermal stability of spray-dried insulin powders compared to commercially available aqueous insulin pharmaceuticals is provided herein. Insulin powders were spray-dried from an aqueous feed solution containing a buffer selected from glycine or histidine (in the range of about 5-20 mM), a surfactant selected from PS20 or PS80 (in the range of about 0.001-0.1% (w / v)), and a sugar / trehalose type disaccharide (dihydrate form) at pH 8.5. Due to the high solids content of the paste combined with the relatively low insulin concentration required in the final formulation (e.g., u100 = 3.5 mg / mL), the excipient concentration in the feed solution was greater than 99% (by weight) of the total. This high excipient-to-insulin ratio allowed the paste composition (solids content of about 55-65% (w / w) (powder concentration of approximately 600-780 mg / mL) and measured density of about 1.1-1.2 g / mL) to have a final insulin content of approximately 3.5 mg / mL, equivalent to u100.

[0199] The Buchi B-290 mini spray dryer used to prepare the powder had an inlet temperature of 140°C, a spray nozzle pressure of 60 (measured from a floating ball flow meter built into the instrument), a liquid feed rate of 10% (approximately 3 mL / min), and an aspirator setting of 90%. The spray dried powder was further dried under vacuum (i.e., secondary drying) to reduce the measured moisture content to approximately less than 1% (w / w) and then stored in glass vials and placed in a stability chamber at 40°C / 75% RH. The chemical stability of the insulin powder was evaluated after 123 days (approximately 4 months) and revealed less than 2% loss in insulin peak purity during the storage period.

[0200] As a comparison, commercially available Humulin® R insulin (in aqueous solution) stored at 40°C in glass vials revealed a purity loss of nearly 7% over one month when measured using the same UHPLC method used for the powder. The USP monograph for insulin injectables is 95-105% of label claim, and indicates that commercially available products fall below their stability specifications within one month at accelerated conditions. Thus, the ability to prepare heat stable spray-dried insulin powders for use in therapeutic pastes could improve the heat stability of currently available commercial formulations while still allowing for comparable injection volumes.

[0201] Example 7: Preparation of high solids paste containing high dose human protein or peptide In further studies, Xeriject paste formulations of high concentration human recombinant proteins were prepared according to the spray drying method described in the previous examples. In the first step, the solution of human recombinant proteins prepared and optimized according to the methods described herein (see Examples 1 and 2) was converted into dry powder by the spray drying process described in the previous examples. Samples of the powder preparations were then examined for morphology and size distribution by scanning electron microscopy. Representative micrographs are shown in Figure 25.

[0202] As discussed in previous examples, the ability to inject high concentrations and large amounts of protein through subcutaneous injection of a relatively small amount of paste formulation requires the formation of small and generally spherical particles of appropriate size in the powder used to prepare therapeutic paste.As seen in Figures 25A and 25B, the protein powder prepared by the spray drying process described herein produces small spherical particles with a relatively small specific surface area per particle.Based on the population statistics (not shown) of the entire powder preparation and the studies described elsewhere herein (see, for example, previous examples), it was determined that the median particle size is optimal for paste formation.

[0203] These powders were then used to prepare Xerreject™ paste formulations using the methods described above, resulting in pastes with a solids content of about 45% and over 300 mg / mL protein in the paste. The therapeutic dose of paste with this concentration of active agent was calculated to be only about 150 μL. The injection force required to deliver this amount of paste prepared in this example at a volumetric flow rate of 30 μL per second in commercially available small and large syringes fitted with either standard or thin-walled 27G needles was then measured as described above. The results of these studies are shown in FIG. 26 and demonstrate that injection forces as low as 6 N are possible with the optimal syringe / needle configuration (i.e., small syringes with thin-walled needles).

[0204] Example 8: Preparation of a high-concentration paste containing bevacizumab In these studies, the Xerreject™ technology was evaluated as a platform for the subcutaneous delivery of small volumes of bevacizumab (marketed by Genentech under the brand Avastin®), a commonly used therapeutic monoclonal antibody formulation that was previously only available for intravenous administration. Bevacizumab is a human monoclonal antibody that binds to vascular endothelial growth factor (VEGF), preventing the interaction of VEGF with its receptor and delaying or preventing angiogenesis, particularly in cancer tissues. As such, it has been approved for a number of indications in humans, including the treatment of metastatic colorectal cancer, non-squamous non-small cell lung cancer, glioblastoma, and metastatic renal cell carcinoma. It is typically administered intravenously (IV) at a dose of 10 mg / kg in an aqueous solution of 25 mg / mL via a 30-90 minute infusion approximately every 2 weeks. The half-life of this product is approximately 20 days, and the mean clearance rate is approximately 0.262 L / day, depending on patient-specific parameters such as weight, sex, and tumor burden. Thus, the inventors evaluated whether Xeriject™ technology could be used to create a formulation of bevacizumab that could be administered in smaller / higher doses and that had at least similar, if not more favorable, pharmacokinetics, both compared to IV products. As described elsewhere herein, such a formulation would provide significant benefits to patients and caregivers, including the use of very small injections, both of which would be less uncomfortable for the patient, but also easier to administer as a subcutaneous dose than via an IV route.

[0205] To conduct these studies, commercially available bevacizumab drug substance was formulated into two different therapeutic paste formulations designated XJ-1 and XJ-2. The compositions (mg / mL) of the aqueous solutions before spray drying and the approximate weight % of the spray-dried powder are shown in Table 24.

[0206] Table 24. Bevacizumab formulations prepared using Xeriject™ technology TIFF2024534963000024.tif79152

[0207] To examine these powders more closely at the particle level, samples of powder from each of these two formulations were examined using SEM to assess particle size, size distribution, and shape (morphology), as discussed in Examples 1-3 above. Representative micrographs are shown in Figure 27, which show that the particles observed in the spray-dried formulations of XJ-1 (Figure 27A) and XJ-2 (Figure 27B) demonstrated a range of spherical particles with an average size of about 3.3 μm (for XJ-1) or about 3.0 μm (for XJ-2). The XJ-2 formulation also showed slightly more donut-shaped particles than the XJ-1 formulation.

[0208] Based on these results, these formulations were used to prepare Xerreject™ formulations of bevacizumab for use in animal pharmacokinetic studies. For each paste, XJ-1 and XJ-2 powders were mixed separately with Miglyol 812 in an HDPE container using a planetary mixer. The solids content of each of the two paste formulations was 62% for XJ-1 and 55% for XJ-2, respectively. The measured mAb content (absorbance at 280 nm) for the two formulations was 429 mg / mL for XJ-1 and 328 mg / mL for XJ-2.

[0209] In these studies, four groups of female Göttingen minipigs were administered one of four different bevacizumab (BmAb) formulations at a single 100 mg dose either intravenously (IV) or subcutaneously (SC).

[0210] Table 25. Bevacizumab PK study design TIFF2024534963000025.tif29162

[0211] After injection of each formulation, plasma was collected from the animals at 5 and 30 minutes, 2, 4, 8, 12 and 24 hours, and 3, 5, 7, 10, 14, 17, 24, 28, 31, 35, 38, 42, 45, 49, 56 and 60 days after injection. Circulating bevacizumab concentrations were then measured in the plasma samples to determine maximum plasma concentrations (T max ) and the time to maximum absorption concentration (C max ), plasma half-life (T1 / 2 ), dose-adjusted exposure (AUC), and fractional exposure (AUC) were assessed. The results of these studies are shown in Figures 28-33.

[0212] As can be seen in FIG. 28, the two Xeriject™ bevacizumab formulations, XJ-1 and XJ-2, exhibited plasma concentration kinetics very similar to the subcutaneously delivered Avastin® formulation, regardless of whether these results were plotted on a linear scale (FIG. 28A) or semi-logarithmic scale (FIG. 28B). Intravenously administered Avastin exhibited the typical bolus effect at early time points commonly seen with IV administered therapeutics. Next, the maximum plasma concentration (T max The time required to reach T ) for each formulation was evaluated, and the results are shown in Figure 29. max was also found to be shorter than the subcutaneously administered Avastin formulations (18 hours for XJ-1 and 24 hours for XJ-2 vs. 72 hours for SC Avastin), but, as expected, was longer than that seen with IV administered Avastin (0.08 hours).

[0213] Next, the maximum absorption (C max ) and evaluate the uncorrected C max values ​​(Figure 30A) or C corrected for dose and body weight of the animals. max The results of the T values ​​(Figure 30B) are shown in Figure 30. These results are consistent with those shown in Figure 29. max Together, these results suggest that the Xerreject formulation of bevacizumab may have a shorter T max (Figure 29) and higher dose-corrected C max (Figure 30B) values, indicating that bevacizumab formulations administered subcutaneously were absorbed more quickly than subcutaneously administered Avastin. Evaluation of the half-life of each of these formulations showed that the subcutaneously administered bevacizumab formulations (both Xeliject and Avastin) demonstrated a shorter half-life than that observed for IV administered Avastin (Figure 31).

[0214] To assess the total exposure of the animals to the antibody, the dose-adjusted exposure (AUC lastand AUC ∞ The results are shown in Figure 32 and show dose-adjusted exposure (AUC last (Figure 32A) and AUC ∞ (Figure 32B)) were similar between the Xerreject and subcutaneously administered Avastin formulations, demonstrating that both assessments showed slightly higher exposure values ​​in animals administered Avastin intravenously. Dose-adjusted 14-day fractional exposure (AUC 0-336hr ; Figure 33), although one of the Xerreject formulations (XJ-2) demonstrated slightly higher fractional exposure values ​​compared to subcutaneously administered Avastin.

[0215] The combined results of these studies, unadjusted for dose and adjusted for dose, are presented in Tables 26 and 27.

[0216] Table 26. Overall Pharmacokinetic Results, Xerreject vs. Avastin (Unadjusted) TIFF2024534963000026.tif36170*T max Values ​​are medians, all other values ​​represent means.

[0217] Table 27. Overall Pharmacokinetic Results, Xerreject vs. Avastin (Dose Corrected) TIFF2024534963000027.tif51170All values ​​represent averages.

[0218] In summary, the results presented in this Example show that bevacizumab plasma concentration-time curves in minipigs are similar for the two different Xeriject™ bevacizumab formulations administered subcutaneously and for Avastin administered subcutaneously, but intravenously administered Avastin has higher peak exposures. Both bevacizumab paste formulations showed shorter T max and significantly higher dose-corrected C max As shown by the values, the absorption of Avastin was faster than that of subcutaneously administered Avastin. Furthermore, the dose-adjusted exposure (AUC last and AUC∞ ) were similar between the Xerreject formulation and subcutaneously administered Avastin. Dose-adjusted partial 2-week exposure (AUC 336 ) tended to be higher for the XJ-2 bevacizumab formulation than for subcutaneously administered Avastin. Finally, the total bioavailability of the subcutaneously administered Avastin and Xeriject bevacizumab formulations was lower than that observed for intravenously administered Avastin, although both could still be within therapeutic dose levels. Thus, the Xeriject technology could be used to prepare highly concentrated formulations of bevacizumab that can be injected subcutaneously in smaller volumes and less frequently than those currently used to deliver Avastin intravenously, while still delivering therapeutic levels of the antibody that appear to be well tolerated and rapidly absorbed in subjects.

[0219] Example 8: Non-clinical evaluation of high solids paste containing insulin To evaluate the pharmacokinetic (PK) and pharmacodynamic (PD) profiles of insulin paste, two paste formulations (XJ-6 and XJ-8) were prepared as described in Example 6 and evaluated in Yucatan minipigs. Both XJ-6 and XJ-8 were formulated to have an insulin content equivalent to u200 (7.5 mg / mL). Insulin powder was prepared by spray drying using a feed solution with total solids (dissolved material) of 50 mg / mL and containing recombinant human insulin (0.52 mg / mL), trehalose (from dihydrate: 49.0 mg / mL), histidine (0.3 mg / mL), PS80 (0.01 mg / mL), EDTA (0.1 mg / mL), and pH adjusted to 9.0 (±0.1) with NaOH and / or HCl.

[0220] The solution was filtered (0.2 μm PVDF membrane) and spray dried in a BUCHI B-290 apparatus with the following settings: inlet temperature = 140 °C / liquid feed rate = 8% / nozzle gas pressure ball meter reading = 60 mm / aspirator setting = 90%. The powder was secondary dried under vacuum to reduce the moisture content of the powder to < 2% (w / w). The powder was mixed with either pure Miglyol 812 (XJ-6) or Miglyol 812 containing 1% (v / v) PS80 (XJ-8). The solids content of XJ-6 and XJ-8 was 57% and 56%, corresponding to a solids concentration of approximately 680 mg / mL and 670 mg / mL, respectively.

[0221] These test products (XJ-6 (Zeriject-6) and XJ-8 (Zeriject-8)) were evaluated for their pharmacokinetic (PK) and pharmacodynamic (PD) properties (changes in blood glucose levels) in Yucatan minipigs and compared with the commercial product Humulin R (u100). Each formulation was subcutaneously injected at a dose of 0.5 U / kg insulin (0.017 mg / kg insulin) into six male Yucatan minipigs.

[0222] The insulin pharmacokinetic profiles of all three formulations were similar, although insulin exposure was lower with XJ-6 as shown in Figure 34. SC administration of 0.5 U / kg Humulin R (3.5 mg / mL insulin) to minipigs resulted in mean (±SD) C max was 13±3ng / mL, and the mean (±SD) AUC last resulted in plasma insulin exposure of 1175±144ng*min / mL. Median T max The mean (±SD) C half-life was 30 min (range: 10 min to 45 min) and the mean (±SD) half-life was 79 ± 12 min. SC administration of 0.5 U / kg XJ-6 (17.4 mg / mL insulin) to minipigs resulted in a mean (±SD) C max was 10±3ng / mL and the mean (±SD) AUC last The insulin exposure was slightly lower than with Humulin R, which had a median t of 902±139ng*min / mL, but similar results were obtained. maxThe mean (±SD) C max was 7.0±1.1ng / mL, and the mean (±SD) AUC last This resulted in lower insulin exposure than Humulin R, which had a median T of 643±92ng*min / mL. max (30 min, range: 20 to 45 min) and half-life (67 min) were similar to that of Humulin R.

[0223] Administration of Humulin R, XJ-6 and XJ-8 produced similar PD responses by rapidly lowering blood glucose levels as shown in Figure 35. Humulin R reduced blood glucose from a mean (±SD) baseline of 72±2 mg / dL to a mean (±SD) of 21±4 mg / dL in a median time of 38 minutes (range: 30-45 minutes). Xereject-6 reduced blood glucose from a mean (±SD) baseline of 78±3 mg / dL to a mean (±SD) of 14±6 mg / dL in a median time of 75 minutes (range: 20-360 minutes). Xereject-8 reduced blood glucose from a mean (±SD) baseline of 78±3 mg / dL to a mean (±SD) of 17±6 mg / dL in a median time of 83 minutes (range: 30-120 minutes). In some cases, hypoglycemia occurred after insulin administration and the animals were treated with oral administration of 50% glucose. The pharmacodynamic effects of the test articles on glucose levels shown here are only approximate for minipigs given glucose for the treatment of hypoglycemia.

[0224] Example 9: Production, characterization and preparation of an injectable paste containing immunoglobulin G (IgG) The following example describes the preparation of an injectable paste formulation containing high concentrations of polyclonal antibody (pAb) immunoglobulin G (IgG). The solid phase in this example comprised an IgG powder prepared by spray drying an aqueous feed solution having a solids content of approximately 51 mg / mL, of which 40 mg / mL was IgG protein (corresponding to 78% (by weight) of the total solids content). Prior to preparation of the final feed solution, the IgG was buffer exchanged with an aqueous solution to obtain the final feed solution composition listed in Table 28.

[0225] Table 28. Formulation feed composition for IgG powder preparation TIFF2024534963000028.tif43164

[0226] This formulation feed was spray dried using a BUCHI B-290 spray dryer parameters and conditions shown below in Table 29.

[0227] Table 29. Spray dryer parameters and conditions for IgG powder preparation TIFF2024534963000029.tif27164

[0228] The spray dried powder was secondary dried under reduced pressure to reduce the moisture content (<1% (w / w)) and then evaluated by scanning electron microscopy (SEM) to examine particle morphology. As can be seen in Figures 36A and 36B, the particles exhibited a generally spherical shape with relatively smooth surfaces, minimal to no observed surface dimples, and a moderately polydisperse size distribution (span approximately 2.0).

[0229] The particle size and particle size distribution of the IgG powder was then determined by laser diffraction (samples were dispersed in a non-solvent (e.g., propyl alcohol) while continuously sonicating the samples to break up powder agglomerations). As shown in Figure 37, the small particle population with D90<10 μm has moderate polydispersity (span about 2) and there is a bimodal distribution with a distinct fine particle (<1 μm) population and a larger particle (1-10 μm) population. It is noted that a relatively bimodal particle size distribution was observed under the conditions described in this example. However, alterations to the formulation and / or process parameters and / or equipment (e.g., larger spray dryers, etc.) and / or characterization methods can shift the observed particle size distribution to a generally unimodal or multimodal (e.g., bimodal, trimodal, etc.) profile that is still suitable for preparing pastes with high solids concentrations.

[0230] Following characterization by SEM, laser diffraction, and moisture content analysis (data not shown), a paste was prepared by mixing (using a planetary mixer) the powder with Miglyol 812N until the solids content was 65%. The density of the powder, measured by helium pycnometer, was approximately 1.2 g / mL, corresponding to a solids concentration of approximately 780 mg / mL. The solids content of the feed solution, specifically the weight percent of protein in the feed solution as a percentage of the total solids (approximately 78% w / w), can be converted to the approximate weight percent of IgG in the spray-dried powder (approximately 78% (w / w)), which can then be used to determine the approximate protein concentration in the paste (approximately 600 mg / mL).

[0231] The IgG paste was also imaged by SEM to determine any changes in particle morphology and / or size distribution (after mixing with Miglyol) and to observe particle packing within the paste. As shown in Figures 38A, 38B, and 38C, the morphology and size distribution of particles comprising the solid phase of the paste remained relatively unchanged after preparation of the IgG paste. SEM analysis of the IgG paste showed good particle packing / arrangement resulting from the high solid phase of the paste, which imparts semi-solid and viscoelastic properties to the paste and may sterically inhibit particle settling over time over pharmaceutical relevant storage conditions and periods.

[0232] Finally, to evaluate the injectability of the high solids concentration IgG paste, 1 mL of paste was loaded into a cyclic olefin copolymer (COC) syringe (Schott) with a 5.0 mm inner barrel diameter and delivered through a 25 gauge, very thin wall, 6 mm needle. Figure 39 provides the injection force profile (plotted as force in Newtons (N; Y axis) against plunger travel distance in millimeters (mm, X axis)) of an IgG paste with a solids content of 65% as measured using a TA.XT Plus model texture analyzer (Stable Micro Systems). The measured injection force (average glide force) was less than 30 N at a volumetric flow rate of 31 μL / sec, as measured by the texture analyzer.

[0233] Thus, this example demonstrates that paste compositions having both high solids concentrations (>700 mg / mL) and high protein concentrations (>500 mg / mL) can be prepared and delivered with modest injection forces through syringes and needles associated with intradermal injection.

[0234] These results, taken together with the preceding examples, demonstrate that paste formulations of high concentration therapeutic proteins provided by the present invention are useful for delivering controlled or sustained release depots of therapeutic proteins, including those of relatively high molecular weight, in small subcutaneous doses in a manner that enhances the patient experience beyond that achieved with larger doses of aqueous formulations of recombinant human proteins.

[0235] The present invention has been described above using functional components that illustrate the implementation of specific functions and their relationships. The boundaries of these functional components are arbitrarily defined in this specification for the convenience of description. Alternative boundaries can be defined as long as the specific functions and their relationships are appropriately implemented.

[0236] The foregoing description of the specific embodiments will sufficiently clarify the basic nature of the invention so that others may easily modify and / or adapt such specific embodiments for various applications without departing from the basic concept of the invention, without undue experimentation, by applying knowledge within the skill of the art. Thus, in addition to those specifically described herein, other suitable embodiments of the invention will be readily apparent to those skilled in the art based on the foregoing description and examples and the knowledge generally available in the relevant technical field. Thus, such adaptations and modifications are intended to be within the meaning and range of equivalents of the disclosed embodiments, based on the teaching and guidance provided herein. It should be understood that the expressions or terms in this specification are for the purpose of description, not limitation, as the terms or terms in this specification will be interpreted by those skilled in the art in light of the teaching and guidance.

[0237] The breadth and scope of the present invention should not be limited by any of the above-described exemplary embodiments, but should be defined only in accordance with the following claims and their equivalents.

[0238] All documents cited herein, including U.S. patents and published patent applications, international patents and patent applications, as well as publications or other publicly available documents, are incorporated by reference in their entirety to the same extent as if the portions of such documents available in the section of this application to which they pertain were specifically cited.

Claims

1. 1. A composition comprising a paste having a solids concentration greater than about 350 mg / mL, the paste comprises one or more active pharmaceutical ingredients, one or more pharmaceutically acceptable excipients, and one or more non-solvent fluids, and the paste can be injected subcutaneously, intradermally, or intramuscularly into an animal in a volume of 3 ml or less at a flow rate of at least 30 μL / s using a commercially available needle / syringe combination; The composition.

2. 10. The composition of claim 1, wherein the paste has a solids concentration of about 350 mg / mL to about 850 mg / mL.

3. 10. The composition of claim 1, wherein the paste has a relative content of active pharmaceutical ingredient of about 20% to about 70%.

4. 10. The composition of claim 1, wherein the active pharmaceutical ingredient is selected from the group consisting of a peptide therapeutic, a protein therapeutic, and a small molecule therapeutic.

5. The composition of claim 4, wherein the peptide or protein therapeutic agent is selected from the group consisting of enzymes, antithrombin agents, thrombolytic agents, peptide hormones, bone-active peptides, diabetes-active peptides, antibodies, non-antibody antitumor agents, pregnancy-promoting agents, and immunosuppressants.

6. The composition of claim 5, wherein the enzyme is selected from the group consisting of dornase alfa, velaglucerase alfa, taliglucerase alfa, asparaginase, glucarpidase, asfotase alfa, elosulfase alfa, sebelipase alfa, sacrosidase, and pegloticase.

7. The composition of claim 5, wherein the antithrombin agent is selected from the group consisting of lepirudin, bivalirudin, defibrotide, and sulodexide.

8. The composition of claim 5, wherein the thrombolytic agent is selected from the group consisting of reteplase, anistreplase, tenecteplase, streptokinase, and urokinase.

9. The composition of claim 5, wherein the peptide hormone is selected from the group consisting of cosinotropin, chorionic gonadotropin, and somatotropin.

10. 6. The composition of claim 5, wherein the bone-active peptide is calcitonin.

11. 6. The composition of claim 5, wherein the diabetes active peptide is selected from the group consisting of insulin, pramlintide, glucagon, and analogs thereof.

12. 12. The composition according to claim 11, wherein the diabetes active peptide is insulin or an analog thereof.

13. 13. The composition of claim 12, wherein the insulin analog is selected from the group consisting of insulin lispro, insulin glargine, insulin aspart, insulin detemir, and insulin glulisine.

14. 12. The composition of claim 11, wherein the diabetes active peptide or protein is glucagon or an analog thereof.

15. 15. The composition of claim 14, wherein the glucagon analog is dasiglucagon.

16. The composition of claim 5, wherein the antibody is a monoclonal antibody or a fragment thereof.

17. 17. The composition of claim 16, wherein the monoclonal antibody is selected from the group consisting of cetuximab, trastuzumab, bevacizumab, rituximab, obinutuzumab, gemtuzumab, canakinumab, ipilimumab, daratumumab, vedolizumab, ustekinumab, siltuximab, ramucirumab, pembrolizumab, ofatumumab, nivolumab, mepolizumab, brodalumab, pertuzumab, denosumab, golimumab, belimumab, raxibacumab, blinatuomab, dinutuximab, and ibritumomab.

18. The composition of claim 5, wherein the non-antibody antitumor agent is selected from the group consisting of leuprolide, denileukin diftitox, aldesleukin, asparaginase, pegaspargase, interferon beta, afibercept, lenograstim, and sipuleucel-T.

19. The composition of claim 5, wherein the immunosuppressant is selected from the group consisting of etanercept, peginterferon alpha, interferon alpha, filgrastim, pegfilgrastim, sargramostim, anakinra, interferon beta, interferon gamma, adalimumab, infliximab, basiliximab, muromonab, efalizumab, daclizumab, abatacept, rilonacept, belatacept, natalizumab, brinzumomab, ustekinumab, and human immunoglobulin.

20. 5. The composition of claim 4, wherein the peptide or protein therapeutic is a recombinant peptide or protein.

21. 5. The composition of claim 4, wherein the small molecule therapeutic is selected from the group consisting of epinephrine or analogs thereof, benzodiazepines, catecholamines, "triptans," novantrones, chemotherapeutic small molecules, corticosteroid small molecules, immunosuppressive small molecules, anti-inflammatory small molecules, small molecules used to treat neurological disorders, small molecules used to treat cancer, statins, taxol and other taxane derivatives, small molecules used to treat tuberculosis, small molecule antifungals, small molecule anxiolytics, small molecule anticonvulsants, small molecule anticholinergics, small molecule beta-agonists, small molecule mast cell stabilizers, small molecule drugs used to treat allergies, small molecule anesthetics / small molecule antiarrhythmics, small molecule antibiotics, small molecule antimigraine drugs, and small molecule antihistamines, and salts or analogs thereof.

22. 22. The composition of claim 21, wherein the small molecule therapeutic agent is a benzodiazepine.

23. 22. The composition of claim 21, wherein the small molecule therapeutic agent is epinephrine or an analog thereof.

24. 10. The composition of claim 1, wherein the one or more pharmaceutically acceptable excipients are selected from the group consisting of sugars, surfactants, amino acids, and buffering agents.

25. 25. The composition of claim 24, wherein the sugar is selected from the group consisting of trehalose, dextrose, sucrose, mannose, and fructose.

26. The composition of claim 24, wherein the surfactant is polysorbate 20 or polysorbate 80.

27. 25. The composition of claim 24, wherein the amino acid is a naturally occurring amino acid selected from the group consisting of proline, cysteine, tryptophan, phenylalanine, arginine, and histidine.

28. 25. The composition of claim 24, wherein the buffering agent is selected from the group consisting of histidine, citrate, succinate, and lactate.

29. The composition of claim 1, wherein the non-solvent fluid is triacetin, Miglyol 810, Miglyol 840, or Miglyol 812.

30. 10. The composition of claim 1 for use in a method for treating, preventing, ameliorating, or diagnosing a disease or disorder in an animal or human suffering from or susceptible to the disease or disorder, the method comprising injecting the composition into the animal or human subcutaneously, intradermally, or intramuscularly.

31. The method comprising the steps of preparing a combination device comprising a needle attached to a syringe, the device containing the composition within the barrel of the syringe in an amount sufficient to deliver a therapeutic dose of at least one active pharmaceutical ingredient to the animal or human; introducing the needle of the syringe-needle combination into a dermal, subcutaneous, or muscular layer of the animal or human; and moving the plunger of the syringe to expel the paste from a reservoir of the syringe through a lumen of a needle attached to the syringe, thereby expelling the paste through the needle and into the animal or human, the reservoir having an internal first transverse dimension that is greater than an internal second transverse dimension of the lumen, the second transverse dimension being between 0.1 and 0.9 mm; Including, the paste has a solids concentration greater than 350 mg / L; and The paste is ejected at a flow rate of more than 30 μL / s.

31. The composition of claim 30.

32. The composition of claim 31, wherein the method includes connecting the needle to the reservoir via a luer fitting disposed on at least one of the needle and the reservoir.

33. 32. The composition of claim 31, wherein the flow rate of the paste is substantially linearly proportional to the speed of movement of the plunger.

34. 32. The composition of claim 31, wherein the first lateral dimension is between 3 and 40 times greater than the second lateral dimension.

35. 32. The composition of claim 31, wherein the first lateral dimension is 1 to 5 mm.

36. 32. The composition of claim 31, wherein the second lateral dimension is 0.1 to 0.9 mm.

37. 32. The composition of claim 31, wherein the needle has a size of 18 gauge or smaller.

38. 38. The composition of claim 37, wherein the needle has a size of 27 gauge.

39. 32. The composition of claim 31, wherein the injection volume of the paste is greater than 10 μL.

40. 40. The composition of claim 39, wherein the injection volume of the paste is 15 μL to 3000 μL.

41. 32. The composition of claim 31, wherein the paste has a solids concentration of 350 mg / mL to 850 mg / mL.

42. 32. The composition of claim 31, wherein the paste has a solids content of 1% to 99%.

43. 32. The composition of claim 31, wherein the paste has a density of 1.0 to 1.5 g / mL.

44. 32. The composition of claim 31, wherein the disease or disorder is selected from the group consisting of a diabetic disease or disorder, an inflammatory disease or disorder, a neurological disease or disorder, cancer, an infectious disease, a bacterial disease, a fungal disease, a viral disease, and a disease, disorder, or condition with inflammatory, neurological, osteological, gastrointestinal, circulatory, cardiovascular, cutaneous, muscular, or developmental signs or symptoms.

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