Dry powder formulations comprising leucine and trileucine

Abstract

To provide dry powder formulations suitable for pulmonary delivery.SOLUTION: Disclosed is a dry powder formulation comprising a plurality of microparticles, where the microparticles comprise: a. leucine; b. about 0.5% to about 10% by weight of trileucine; and c. an active agent, where the weight ratio of the leucine to the trileucine is about 0.1:1 to about 30:1.SELECTED DRAWING: Figure 1

Description

[Technical Field]

[0001] The present technology relates generally to dry powder formulations suitable for pulmonary delivery. Also provided are methods of preparing the dry powder formulations, and methods of administration and treatment using the dry powder formulations. [Background technology]

[0002] Advantages of pulmonary delivery of active agents include the convenience of patient self-administration, the potential for reduced drug side effects, ease of delivery by inhalation, the elimination of needles, and the like. Numerous clinical studies involving the inhalation of proteins, peptides, DNA, and small molecules have demonstrated that efficacy can be achieved both in the lungs and systemically. However, many molecules (especially biomolecules) that require high payloads for delivery present challenges in developing inhalable formulations. The formulation must provide stability to the biological payload and be scalable for manufacturing, while also maintaining the desired physical properties to facilitate delivery into the patient's lungs. Summary of the Invention [Means for solving the problem]

[0003] In view of the above, provided herein is a dry powder formulation comprising a plurality of microparticles, the microparticles comprising leucine; about 0.5% to about 10% by weight of trileucine; and an active agent, wherein the leucine and trileucine are present in a ratio of leucine:trileucine of about 0.1:1 to about 30:1 by weight. In certain embodiments, the dry powder formulation has a compressed bulk density of about 0.4 to 1.0 g / cm. 3 is.

[0004] Also provided herein is a method for preparing a dry powder formulation, comprising: preparing a liquid feedstock comprising leucine; about 0.1 mg / mL to about 6 mg / mL trileucine; an active agent; and a liquid solvent, wherein the leucine and trileucine are present in a concentration ratio of leucine:trileucine of about 0.1:1 to about 30:1; atomizing the liquid feedstock; and drying the atomized liquid feedstock to form a plurality of microparticles.

[0005] In a further embodiment, provided herein is a method of preparing a dry powder formulation comprising a plurality of particulates having a compressed bulk density of about 0.4 to about 1.0 g / cm, the method comprising incorporating into the dry powder formulation leucine and trileucine in a ratio of leucine:trileucine of about 0.1:1 to about 30:1 by weight.

[0006] Also provided herein is a polymer having a specific surface area of ​​about 5 to about 10 m 2 1. A method for preparing a dry powder formulation comprising a plurality of microparticles, each microparticle having a particle size of about 1 / g, the method comprising incorporating leucine and trileucine into the dry powder formulation in a ratio of leucine:trileucine of about 0.1:1 to about 30:1 by weight.

[0007] In a further embodiment, provided herein is a method of preparing a dry powder formulation comprising a plurality of microparticles, the microparticles having a mass median aerodynamic diameter (MMAD) of from about 2 μm to about 4 μm when provided in aerosol form, the method comprising incorporating leucine and trileucine into the dry powder formulation in a ratio of leucine:trileucine of from about 0.1:1 to about 30:1 by weight.

[0008] In a further embodiment, provided herein is a method of delivery of a dry powder formulation to the lungs of a mammalian patient, the method comprising administering to the mammalian patient by inhalation an aerosol form of a dry powder formulation provided herein.

[0009] Also provided herein is a method of treating a medical condition in a mammalian patient, comprising administering to the mammalian patient by inhalation an aerosol form of a dry powder formulation described herein.

[0010] In additional embodiments, the dry powder formulations described herein may be used in the methods of treatment, where the formulations are administered by inhalation.

[0011] The above and other features and aspects of the present technology may be better understood from the following description of embodiments and as illustrated in the accompanying drawings, which are incorporated in and constitute a part of this specification and further serve to explain the principles of the present technology. The drawings are not necessarily to scale. [Brief explanation of the drawings]

[0012] [Figure 1] 1 shows microparticles from a dry powder formulation according to an embodiment. [Figure 2A] 1 shows the results of compacted bulk density as a function of leucine and trileucine in dry powder formulations. [Figure 2B] 1 shows the filling of capsules with the dry powder formulation described herein. [Figure 3] 1 shows the specific surface area results (m / g) measured using BET for particulates of dry powder formulations according to embodiments. [Figure 4] 1 shows an indirect correlation between moisture content and leucine concentration. [Figure 5A] The surface roughness of the particles as detected by DSEM is shown. [Figure 5B] The surface roughness of the particles as detected by DSEM is shown. [Figure 5C] The surface roughness of the particles as detected by DSEM is shown. [Figure 5D] Figure 5A shows the surface roughness of the fine particles as detected by DSEM. [Figure 6]1 shows the correlation between fine particle fraction (FPF) and wt% of leucine and trileucine. [Figure 7] 1 shows the correlation between device deposition and wt% values ​​of leucine and trileucine. [Figure 8] 1 shows the correlation between fine particle fraction (FPF) and wt% values ​​of leucine and trileucine. [Figure 9A] 1 shows the saturation velocities modeled at various leucine and trileucine concentration combinations. [Figure 9B] 1 shows the saturation velocities modeled at various leucine and trileucine concentration combinations. [Figure 10A] The number of subvisible particles after reconstitution of formulations containing 40% (w / w) Fab1 and various concentrations of polysorbate-80 (PS-80) to a solution concentration of 30 mg / ml of Fab1 is shown (in the figure, "≧" includes an upper size limit of 200 μm). [Figure 10B] The number of subvisible particles after reconstitution of formulations containing 40% (w / w) Fab1 and various concentrations of PS-80 to a solution concentration of 2.5 mg / ml of Fab1 is shown (in the figure, "≧" includes an upper size limit of 200 μm). [Figure 11A] The number of subvisible particles after reconstitution of formulations containing 40% (w / w) Fab1 and various concentrations of poloxamer-188 to a solution concentration of 30 mg / ml Fab1 is shown (in the figure, "≧" includes an upper size limit of 200 μm). [Figure 11B] The number of subvisible particles after reconstitution of formulations containing 40% (w / w) Fab1 and various concentrations of poloxamer-188 to a solution concentration of 2.5 mg / ml of Fab1 is shown (in the figure, "≧" includes an upper size limit of 200 μm). [Figure 12A] Figure 1 shows the % moisture content of a formulation containing 40% (w / w) Fab1 and 1.1% PS-80 after storage for 1 month and 3 months at 40°C and 75% relative humidity (40 / 75) and 3 months at 25°C and 60% relative humidity (25 / 60). [Figure 12B]Figure 1 shows the particle size distribution (PSD) of a formulation containing 40% (w / w) Fab1 and 1.1% PS-80 after storage for 1 month and 3 months at 40°C and 75% relative humidity (40 / 75) and 3 months at 25°C and 60% relative humidity (25 / 60). [Figure 12C] FIG. 1 shows the particle morphology of a formulation containing 40% (w / w) Fab1 and 1.1% PS-80 after storage for 1 month and 3 months at 40° C. and 75% relative humidity (40 / 75) and 3 months at 25° C. and 60% relative humidity (25 / 60). [Figure 13A] Figure 1 shows the % moisture content of a formulation containing 1% (w / w) Fab1 and 1.1% PS-80 after storage for 1 month or 3 months at 40°C and 75% relative humidity (40 / 75) and 3 months at 25°C and 60% relative humidity (25 / 60). [Figure 13B] Figure 1 shows the particle size distribution (PSD) of a formulation containing 1% (w / w) Fab1 and 1.1% PS-80 after storage for 1 or 3 months at 40°C and 75% relative humidity (40 / 75) and 3 months at 25°C and 60% relative humidity (25 / 60). [Figure 13C] FIG. 1 shows the particle morphology of a formulation containing 1% (w / w) Fab1 and 1.1% PS-80 after storage for 1 or 3 months at 40° C. and 75% relative humidity (40 / 75) and 3 months at 25° C. and 60% relative humidity (25 / 60). [Figure 14A] Figure 1 shows the number of subvisible particles after reconstitution of a formulation containing 40% Fab1 and 1.1% PS-80 (w / w) to a solution concentration of 30 mg / ml Fab1 after storage at 40 / 75 for 1 month or 3 months and at 25 / 60 for 3 months. [Figure 14B] Figure 1 shows the number of subvisible particles after reconstitution of a formulation containing 1% Fab1 and 1.1% PS-80 (w / w) to a solution concentration of 0.75 mg / ml of Fab1 after storage at 40 / 75 for 1 month or 3 months and at 25 / 60 for 3 months. DETAILED DESCRIPTION OF THE INVENTION

[0013] It should be understood that the specific implementations shown and described herein are examples and are not intended to otherwise limit the scope of the application in any way.

[0014] Published patents, patent applications, websites, company names, and scientific literature referred to herein are incorporated by reference in their entirety to the same extent as if each were specifically and individually indicated to be incorporated by reference. Any conflict between any reference cited herein and the specific teachings of this specification shall be resolved in favor of the latter. Similarly, any conflict between an art-understood definition of a word or phrase and a definition of a word or phrase specifically taught herein shall be resolved in favor of the latter.

[0015] As used herein, the singular forms "a," "an," and "the" include the plural of the terms they specifically refer to, unless the context clearly dictates otherwise. The term "about" is used herein to mean approximately, in the region of, roughly, or approximately. When the term "about" is used in conjunction with a numerical range, it modifies that range by extending the limits above and below the stated numerical values. In general, the term "about" is used herein to modify a numerical value by a variance of 10% above and below the stated value.

[0016] Unless otherwise defined, technical and scientific terms used herein have the meaning commonly understood by one of ordinary skill in the art to which this application belongs. This specification refers to various methods and materials known to those skilled in the art.

[0017] As described herein, dry powder formulations are provided for the stabilization and delivery of pharmaceutically active agents. Preferably, the dry powder formulations are formulated for pulmonary delivery (e.g., by inhalation via a dry powder inhaler (DPI)).

[0018] As used herein, a "dry powder formulation" refers to a formulation comprising a plurality of solid particulates in a powder composition preferably containing less than about 20% moisture, more preferably less than 10% moisture, less than about 5-6% moisture, or less than about 3% moisture. As described herein, dry powder formulations may be utilized for delivery to a patient via inhalation. In other embodiments, dry powder formulations may be reconstituted into a liquid form and administered orally, intravenously, parenterally, or the like. As described herein, an advantage of the provided dry powder formulations is increased throughput for improved manufacturability. An additional advantage is that the formulation platforms described herein offer high compacted bulk densities, meaning that a greater mass of powder can be packaged per delivery unit (e.g., in a capsule). This means that a higher dose of active agent can be delivered to a subject per delivery unit. This surprising advantage can improve patient compliance by reducing the number of unit doses that need to be taken. Additionally, high compacted bulk densities can enable the delivery of higher doses of active agent, increasing the upper end of the administered dose range. This may allow delivery of active agents in therapeutically effective doses that were not previously possible.

[0019] "Microparticles," as used herein, refer to solid particles having a size mass mean diameter (MMD) of less than 20 μm. Mass mean diameter is a measure of the average particle size of the microparticles and is measured using a suitable method (e.g., centrifugal sedimentation, electron microscopy, light scattering, laser diffraction, etc.).

[0020] The dry powder formulations described herein preferably comprise a plurality of microparticles. As used herein, "plurality" refers to two or more items, preferably 5 or more, 10 or more, 50 or more, 100 or more, 500 or more, 1000 or more, etc.

[0021] In embodiments, the dry powder formulation comprises a plurality of microparticles, the microparticles preferably comprising leucine; about 0.5% to about 10% by weight trileucine; and an active agent. Figure 1 shows a scanning electron micrograph of microparticles of an exemplary dry powder formulation provided herein. In further embodiments, the dry powder formulation comprising a plurality of microparticles preferably comprises about 1% to about 25% leucine; about 1% to about 10% trileucine; and an active agent.

[0022] As used herein, "leucine" refers to the amino acid leucine (CH), whether present as a single amino acid or as an amino acid component of a peptide. 13 NO2), the amino acid leucine can be a racemic mixture or in either its D- or L-form, and can be modified forms of leucine (i.e., one or more atoms of leucine have been replaced with another atom or functional group). The chemical structure of leucine is shown below: [ka]

[0023] "Tri-leucine," as used herein, refers to a group of three leucine molecules, leucine-leucine-leucine (Leu-Leu-Leu), C 18 H 35 It refers to compounds that are linked together in a peptide-like manner as N3O4. The chemical structure of trileucine is shown below. [ka]

[0024] The amounts of leucine and trileucine provided herein are given as weight percentages (wt%) of the formulation unless otherwise stated. Because the dry powder formulations contain substantially little, if any, water, the weight components of the dry powder formulations are dry weight percentages of the final formulation.

[0025] In embodiments of formulations comprising leucine, trileucine, and an active agent, the leucine and trileucine are maintained in a desired ratio range that provides the improved compacted bulk density characteristics described herein, as well as desirable microparticle properties that allow for improved storage and delivery. In embodiments, the weight ratio of leucine to trileucine (i.e., leucine:trileucine) in the microparticles is from about 0.05:1 to about 40:1, and more preferably, the leucine and trileucine are present in a weight ratio of leucine:trileucine of from about 0.1:1 to about 30:1. In further embodiments, the leucine and trileucine are present in a weight ratio of leucine:trileucine of about 0.1:1 to about 25:1, about 0.5:1 to about 20:1, about 1:1 to about 20:1, about 1:1 to about 15:1, about 1:1 to about 12:1, about 1:1 to about 10:1, about 1:1 to about 7:1, about 1:1 to about 6:1, or about 1:1, about 2:1, about 3:1, about 4:1, about 5:1, about 5.1:1, about 5.2:1, about 5.25:1, about 5.3:1, about 5.4:1, about 5.5:1, about 5.75:1, or about 6:1.

[0026] Unless otherwise stated, the ratios described herein are expressed as weight percent ratios (w / w—also referred to as “weight ratios”), i.e., the weight of leucine:weight of trileucine in the formulations described herein. This ratio is achieved by providing the desired mg / mL concentrations of leucine and trileucine in the feedstock, and then drying to remove the feedstock solvent to obtain atomized microparticles in which the starting concentration ratio (expressed in mg / mL) is maintained as the final ratio of leucine:trileucine by weight.

[0027] Exemplary weight percentages of leucine and trileucine that can be used in the dry powder formulation to achieve this ratio are described herein. Preferably, the dry powder formulation contains about 5% to about 15% leucine and about 1% to about 5% trileucine. In embodiments, the dry powder formulation contains about 8% to about 11% leucine and about 2% to about 4% trileucine, and in embodiments, the dry powder formulation contains about 10.5% leucine and about 2% trileucine.

[0028] In exemplary embodiments, the dry powder formulation comprises about 0.5% to about 10% trileucine by weight, and more preferably about 1% to about 10%, 1% to about 9%, about 1% to about 8%, about 1% to about 7%, about 1% to about 6%, about 1% to about 5%, about 2% to about 10%, about 2% to about 9%, about 2% to about 8%, about 2% to about 7%, about 2% to about 6%, about 2% to about 5%, about 2% to about 4%, or about 1%, about 1.5%, about 2%, about 2.5%, about 3%, about 3.5%, about 4%, about 4.5%, about 5%, about 5.5%, or about 6% trileucine by weight.

[0029] In exemplary embodiments, the dry powder formulation comprises about 1% to about 25% leucine by weight, and more preferably about 2% to about 20%, about 3% to about 20%, about 4% to about 20%, about 5% to about 20%, about 5% to about 15%, about 7% to about 12%, about 8% to about 11%, about 9% to about 11%, about 10% to about 11%, or about 5%, about 6%, about 7%, about 8%, about 8.5%, about 9%, about 10%, about 10.5%, about 11%, about 11.5%, about 12%, about 12.5%, or about 13% leucine by weight.

[0030] In a preferred embodiment, the dry powder formulation contains about 8% to about 11% leucine and about 2% to about 4% trileucine by weight, and more preferably about 9% to about 11% leucine and about 2% to about 3% trileucine by weight. In an exemplary embodiment, the dry powder formulation contains about 10.5% leucine and about 2% trileucine by weight.

[0031] As described herein, it has been surprisingly discovered that the use of a combination of leucine and trileucine in dry powder formulations can reduce the total amount of leucine and trileucine required to prepare microparticles, compared with dry powder formulations that contain only one of these components, and still provide desired stability.In certain embodiments, the formulations described herein have increased compacted bulk density compared with formulations in the art, which can allow for the delivery of higher concentrations of active agent to patient's lungs after inhalation.This improvement in properties is believed to be related to the incorporation of leucine and trileucine into microparticles.

[0032] An exemplary process for preparing an embodiment dry powder formulation may be as follows: Liquid ingredients containing the desired final components of the dry powder formulation are sprayed into a fine mist using an atomizer. The mist is then dried as described herein. The sprayed droplets initially contain the dissolved components as liquid droplets. As the droplets dry, the various components of the formulation become saturated and begin to precipitate at different rates. As described herein, a shell begins to form around the outer surface of the microparticles of the dry powder formulation. The shell preferably contains leucine and trileucine components on the outer surface of the shell. It should be noted that while leucine and trileucine will be preferentially located on the outer surface of the microparticles, small amounts of leucine and trileucine may also be found throughout the microparticles. In embodiments, higher concentrations of leucine and trileucine are preferably found at or near the surface of the microparticles rather than near the center of the microparticles. In embodiments, the center of the microparticles contains a significant amount of the active agent, preferably in amorphous form, along with other additive components as described herein. As used herein, a "substantial amount" of active agent means that at least about 60% of the active agent (i.e., the total active agent in the formulation) is located at or near the center of the microparticle, preferably at least about 70% of the active agent, more preferably at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, and in one embodiment about 95% to 100% of the active agent is located at or near the center of the microparticle.

[0033] In further embodiments, the microparticles comprise leucine and trileucine located substantially throughout the microparticle, but in greater amounts at or near the surface of the microparticle. As used herein, "substantially throughout the microparticle" means that the leucine and / or trileucine is located in a gradient from the outer surface of the microparticle toward the center of the microparticle, preferably with a decreasing amount of leucine and / or trileucine toward the center, in embodiments where no leucine or trileucine is found in the center of the microparticle, but where the active agent is located. In other embodiments, the amount of leucine and trileucine can be substantially uniform across the cross-section of the microparticle.

[0034] In embodiments, substantially each of the microparticles in the dry powder formulation comprises leucine and trileucine. That is, preferably, at least about 60% of the microparticles comprise leucine and trileucine, or at least about 70%, more preferably at least 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, and in embodiments, about 95% to 100% of the microparticles comprise leucine and trileucine. In embodiments, each of the microparticles in the dry powder formulation comprises leucine and trileucine.

[0035] In additional embodiments, leucine and / or trileucine may be found in the dry powder formulation, but not contained in or associated with the microparticles of the formulation. Thus, in embodiments, free leucine and / or free trileucine not associated with microparticles may be found in the dry powder formulation. However, generally, the amount of free leucine and / or free trileucine (i.e., leucine and / or trileucine not associated with microparticles) is on the order of less than about 10%, less than about 5%, less than about 1%, and more preferably less than about 0.1% of the total amount of leucine and / or trileucine in the formulation.

[0036] In exemplary embodiments, the dry powder formulations described herein have a compressed bulk density that allows for the delivery of a large amount of active agent. "Compressed bulk density" refers to the mass per unit volume of a powder (preferably g / cm) when measured under the following conditions: 3) refers to the compressed bulk density (cBD or CBD). A suitable assay for measuring compressed bulk density (cBD or CBD) is described in the Examples (see, e.g., Example 1). Preferably, the compressed bulk density (CBD) of a powder is measured using a density analyzer (e.g., a GeoPyc® Model 1360 Density Analyzer (Micromeritics, Norcross, GA). A powder sample is preferably prepared in a low humidity environment (<5% RH) and then transferred to the sample chamber of the density analyzer, which is purged with nitrogen gas. The net weight of the powder sample is recorded, and then a compressive force of 10-14 N (preferably 12 N) is applied to the sample by a plunger at a rate of 250-350 compaction strokes per second (preferably 300 compaction strokes per second). The linear distance traveled by the plunger during each compaction stroke is converted to the volumetric displacement of the powder sample. The measurements from each compaction stroke are then averaged to produce a calculated bulk density value (g / cm) for the dry powder formulation. 3 (expressed as

[0037] In certain embodiments, the dry powder formulations described herein have a compressed bulk density of at least 0.4 g / cm 3 and preferably about 0.4 g / cm 3 ~Approx. 1.0g / cm 3 and more preferably about 0.4 to 0.9 gm / cm 3 , about 0.4~0.8gm / cm 3 , about 0.5~0.8gm / cm 3 , about 0.6~0.8gm / cm 3 , or about 0.4 gm / cm 3 , about 0.5gm / cm 3 , about 0.6gm / cm 3 , about 0.7gm / cm 3 , or about 0.8 gm / cm 3 In certain embodiments, the compressed bulk density of the dry powder formulations described herein is about 0.4 gm / cm 3 ~about 0.9gm / cm 3 In certain embodiments, the compressed bulk density of the dry powder formulations described herein is about 0.5 gm / cm 3 ~about 0.8gm / cm3 is.

[0038] FIG. 2A shows the results of compacted bulk density as a function of leucine and trileucine in the dry powder formulations described herein. Each column represents the amount of trileucine in the formulation. Within each column, the amount of leucine increases from about 1% to about 20%. As shown, increasing the amount of trileucine results in lower compacted bulk density, and increasing the amount of leucine within each group also decreases the compacted bulk density. Approximately 0.5 g / cm 3 ~about 0.8g / cm 3 To achieve a compacted bulk density of 1000 mg / kg, the amount of trileucine should be kept below 4% by weight.

[0039] A variety of active agents may be formulated into the dry powder formulations described herein. As used herein, "active agent" refers to a pharmaceutically active organic or inorganic compound that acts on a desired target in a mammalian patient, such as a human, to treat, ameliorate, ameliorate, or cure a symptom, disease, infection, condition, etc., of the patient.

[0040] The amount of active agent contained in the dry powder formulation is preferably in the range of about 10% to 80% by weight, more preferably about 20% to 70%, about 30% to 50%, or about 30%, about 35%, about 40%, about 45%, about 50%, about 55%, about 60%, etc.

[0041] Exemplary active agents include small molecules. As used herein, the term "small molecule" refers to chemically synthesized low-molecular-weight pharmaceutical, therapeutic, and / or diagnostic agents (examples of the latter are markers, dyes, etc.) generally having a molecular weight of less than about 10 kD, preferably less than about 5,000 daltons, more preferably less than about 1,000 daltons, e.g., about 100 to about 900 daltons, about 200 to about 800 daltons, about 300 to about 700 daltons, about 400 to about 600 daltons, or about 500 daltons, as well as salts, esters, and other pharmaceutically acceptable forms of such compounds.

[0042] In additional embodiments, the active agent may be a biologic. As used herein, "biologic" refers to isolated or synthetically produced naturally occurring products (e.g., nucleic acids, amino acids, peptides, polypeptides, and proteins), preferably including antibodies, antigen-binding fragments, and the like.

[0043] The term "polypeptide" refers to a molecule comprising a polymer of amino acids linked together by peptide bonds. Polypeptides include polypeptides of any length, such as proteins (e.g., proteins having more than 50 amino acids) and peptides (e.g., 2-49 amino acids). Polypeptides include proteins and / or peptides of any activity, function, or size, and include secreted, membrane-anchored, or intracellular proteins.

[0044] Exemplary polypeptides and recombinant polypeptides include enzymes (e.g., proteases, kinases, phosphatases), receptors, transporters, bactericidal and / or endotoxin-binding proteins, structural polypeptides, membrane-bound polypeptides, glycoproteins, globular proteins, immune polypeptides, toxins, antibiotics, hormones, growth factors, blood factors, vaccines, or the like. Polypeptides can be peptide hormones, interleukins, tissue plasminogen activators, cytokines, immunoglobulins (including antibodies or functional antigen-binding fragments or variants thereof), and Fc fusion proteins. Polypeptides can also be subunits or domains of polypeptides, such as the heavy or light chains of antibodies, or functional fragments or derivatives thereof.

[0045] In embodiments, the polypeptide is an immunoglobulin molecule, preferably an antibody or a subunit or domain thereof (e.g., an antibody heavy or light chain). The term "antibody," as used herein, refers to a protein comprising at least two heavy chains and two light chains connected by disulfide bonds. The term "antibody" includes naturally occurring antibodies as well as all recombinant antibodies (e.g., humanized antibodies, fully human antibodies, and chimeric antibodies). Each heavy chain typically consists of a heavy chain variable region (VH) and a heavy chain constant region (CH). Each light chain typically consists of a light chain variable region (VL) and a light chain constant region (CL). However, the term "antibody" also includes other types of antibodies (e.g., single-domain antibodies, heavy-chain antibodies, i.e., antibodies composed only of one or more (especially two) heavy chains, and nanobodies, i.e., antibodies composed only of a single monomeric variable domain). Examples of antibody fragments or derivatives include: (i) a Fab fragment, which is a monovalent fragment consisting of the variable regions of each heavy and light chain and the first constant domain; (ii) an F(ab)2 fragment, which is a bivalent fragment comprising two Fab fragments linked by disulfide bridges at the hinge regions; (iii) an Fd fragment consisting of a heavy chain variable region and the first constant domain, CH1; (iv) an Fv fragment consisting of the heavy and light chain variable regions of a single arm of an antibody; (v) an scFv fragment, which is an Fv fragment consisting of a single polypeptide chain; (vi) an (Fv)2 fragment consisting of two Fv fragments covalently linked to each other; (vii) a heavy chain variable domain; and (viii) a multibody consisting of a heavy chain variable region and a light chain variable region that are covalently linked to each other such that binding between the heavy chain variable region and the light chain variable region can occur only intermolecularly, not intramolecularly.

[0046] Examples of active agents include small molecules or biologics useful in treating patients suffering from or susceptible to any disease state, including, but not limited to, cancer (e.g., breast cancer, uterine cancer, ovarian cancer, prostate cancer, testicular cancer, lung cancer, leukemia, lymphoma, colon cancer, gastrointestinal cancer, pancreatic cancer, bladder cancer, renal cancer, bone cancer, neurological cancer, head and neck cancer, skin cancer, sarcoma, adenoma, carcinoma, and myeloma); infectious diseases (e.g., bacterial diseases, fungal diseases, parasitic diseases, and viral diseases (e.g., viral hepatitis, diseases caused by cardioactive viruses; HIV / AIDS, flu, SARS, and the like)); genetic disorders (e.g., For example, anemia, neutropenia, thrombocytopenia, hemophilia, dwarfism, and severe combined immunodeficiency ("SCID"); inflammatory and autoimmune disorders (e.g., psoriasis, systemic lupus erythematosus, and rheumatoid arthritis, asthma, e.g., severe, moderate, or mild asthma, chronic obstructive pulmonary disease, atopic dermatitis, and idiopathic pulmonary fibrosis), and neurodegenerative disorders (e.g., various forms and stages of multiple sclerosis, Creutzfeldt-Jakob disease, Alzheimer's disease, and the like). In certain embodiments, the active agent is for use in the treatment of severe asthma, such as eosinophilic or non-eosinophilic asthma, optionally hypoeosinophilic asthma.

[0047] Exemplary active agents that may be included in the dry powder formulations described herein include, but are not limited to, inhaled corticosteroids (ICS), long-acting beta-agonists (LABAs), leukotriene receptor antagonists (LTRAs), long-acting antimuscarinics (LAMAs), chromones, short-acting beta-agonists (SABAs), cytokines such as interleukins, hormones, interferons, tissue growth factors, endothelial growth factors, phosphodiesterase (PDE) compounds, VLA-4 inhibitors, bisphosphonates, macrolides, antibiotics, fluoroquinolones, aminoglycosides, polymyxins, antifungals, carbapenems, and the like, as well as analogs, agonists, antagonists, inhibitors, and pharmaceutically acceptable salt forms of the above, where applicable.

[0048] With respect to peptides and proteins, it is intended to encompass synthetic, natural, glycosylated, non-glycosylated, and pegylated forms, as well as biologically active fragments and analogs thereof. Active agents also include: nucleic acids as naked nucleic acid molecules, vectors, related viral particles, plasmid DNA or RNA, or other nucleic acid constructs of a type suitable for transfection or transformation of cells (i.e., suitable for gene therapy), such as antisense, siRNA, miRNA, etc. Additionally, active agents may include live, attenuated, or killed viruses suitable for use as vaccines.

[0049] In embodiments, the active agent is an anti-TSLP antibody or antibody variant, preferably an anti-TSLP antigen-binding fragment thereof. Thymic stromal lymphopoietin (TSLP) is an epithelial cell-derived cytokine involved in the initiation of allergic inflammation. The anti-TSLP antigen-binding fragments (designated Fab1) described herein may be useful in the treatment of asthma. Exemplary Fab1 sequences include:

[0050] HCDR1 FAB1 Thr Tyr Gly Met His (SEQ ID NO: 1)

[0051] HCDR2 FAB1 Val Ile Trp Tyr Asp Gly Ser Asn Lys His Ala Tyr Asp Ser Val Lys Gly (SEQ ID NO: 2)

[0052] HCDR3 FAB1 Ala Pro Gln Trp Glu Leu Val His Glu Ala Phe Asp Ile (SEQ ID NO: 3)

[0053] Heavy chain VH FAB1 [ka]

[0054] LCDR1 FAB1 Gly Gly Asn Asn Leu Gly Ser Lys Ser Val His (SEQ ID NO: 5)

[0055] LCDR2 FAB1 Asp Asp Ser Asp Arg Pro Ser (SEQ ID NO: 6)

[0056] LCDR3 FAB1 Gln Val Trp Asp Ser Ser Ser Asp His Val Val (SEQ ID NO: 7)

[0057] Light chain VL FAB1 [ka]

[0058] FAB1 variable heavy chain [ka]

[0059] FAB1 variable light chain [ka]

[0060] In preferred embodiments, the dry powder formulations described herein further comprise a glass stabilizer, which aids in stabilizing the formulation (particularly the active agent). A "glass stabilizer" refers to an additive that stabilizes an active agent (preferably a polypeptide) in a dry powder formulation, preferably by displacing water on the surface of the active agent during drying or otherwise preventing degradation processes, thereby forming an amorphous solid containing the active agent. Examples of glass stabilizers include amorphous sugars, polymeric sugars, buffers, salts, or synthetic polymers (e.g., poly-L-glycolic acid), as well as mixtures of such compounds. In preferred embodiments, the glass stabilizer is an amorphous sugar. In additional embodiments, the glass stabilizer is a buffer. In further embodiments, the formulations described herein may include both an amorphous sugar and a buffer, which may act together or separately as glass stabilizers.

[0061] Exemplary amorphous sugars for use in the formulations described herein include, but are not limited to, trehalose, sucrose, raffinose, inulin, dextran, mannitol, and cyclodextrin. Preferably, the amorphous sugar is present at about 30% to about 70% (by weight) of the dry powder formulation. In further embodiments, the amorphous sugar is present at about 30% to about 65%, about 35% to about 65%, about 35% to about 60%, about 40% to about 60%, about 30% to about 50%, or about 30%, about 35%, about 40%, about 45%, about 50%, about 55%, or about 60%. Preferably, the amorphous sugar is trehalose and is present in the dry powder formulation at about 30% to 60% by weight of the formulation, more preferably at about 35% to 55%, or about 35%, about 40%, about 45%, or about 50%.

[0062] Exemplary buffers that may be included in the dry powder formulations (preferably as glass stabilizers) include various citrate buffers (e.g., sodium citrate), phosphate buffers, histidine buffers, glycine buffers, acetate buffers, and tartrate buffers, as well as combinations of such buffers. The amount of buffer that may be included in the dry powder formulations may range from about 0.1% to about 20%, more preferably from about 0.5% to about 15%, from about 1% to about 10%, from about 2% to about 8%, from about 3% to about 7%, or about 1%, about 2%, about 3%, about 4%, about 5%, about 6%, about 7%, about 8%, about 9%, or about 10%.

[0063] The buffer also controls the pH of the dry powder formulation, preferably maintaining a pH of about pH 5 to about 8 (e.g., about pH 5 to about pH 6, about pH 5.5 to about pH 6.5, about pH 6 to about pH 7, about pH 6.5 to about pH 7.5, or about pH 7 to about pH 8).

[0064] In an additional embodiment, a dry powder formulation is provided comprising about 30%-50% trehalose, about 10%-11% leucine, about 1%-3% trileucine, about 8%-9% citrate buffer, and an active agent, more preferably about 39% trehalose, about 10.5% leucine, about 2% trileucine, about 8.5% citrate buffer, and an active agent.

[0065] In additional embodiments, a dry powder formulation is provided consisting essentially of about 30%-50% amorphous sugar, leucine, about 0.5%-10% trileucine, about 1%-10% buffer, and an active agent, wherein the leucine and trileucine are present in a concentration ratio of about 0.1:1 to about 30:1 leucine:trileucine. In additional embodiments, a dry powder formulation is provided consisting essentially of about 30%-50% amorphous sugar, about 8%-11% leucine, about 2%-4% trileucine, about 1%-10% buffer, and an active agent. An additional dry powder formulation is provided consisting essentially of about 35%-45% trehalose, about 9%-11% leucine, about 2%-3% trileucine, about 2%-85% citrate buffer, and an active agent. In a further embodiment, the dry powder formulation consists essentially of about 39% trehalose, about 10.5% leucine, about 2% trileucine, about 8.5% citrate buffer, and the active agent.

[0066] For compositions and formulations that "consist essentially" of the recited ingredients, such compositions and formulations include the recited ingredients plus those that do not substantially affect the basic and novel properties of the claimed formulation. Ingredients that do not substantially affect the basic and novel properties of the claimed formulation are those that do not limit the ability of leucine and trileucine to stabilize the dry powder formulations. Preferably, compositions and formulations that consist essentially of the recited ingredients specifically exclude other amino acids or tripeptide amino acids, but may include additional sugars, buffers, etc.

[0067] In an exemplary embodiment, a dry powder formulation is provided that contains about 30-50% trehalose, about 10-11% leucine, about 1-3% trileucine, about 8-9% citrate buffer, and about 30-50% anti-TSLP antibody antigen-binding fragment, more preferably about 39% trehalose, about 10.5% leucine, about 2% trileucine, about 8.5% citrate buffer, and about 40% anti-TSLP antibody antigen-binding fragment. In a specific embodiment, the anti-TSLP antibody antigen-binding fragment is Fab1.

[0068] In a further exemplary embodiment, a dry powder formulation is provided that consists essentially of about 30-50% trehalose, about 10-11% leucine, about 1-3% trileucine, about 8-9% citrate buffer, and about 30-50% anti-TSLP antibody antigen-binding fragment, more preferably about 39% trehalose, about 10.5% leucine, about 2% trileucine, about 8.5% citrate buffer, and about 40% anti-TSLP antibody antigen-binding fragment.

[0069] The microparticles comprising the dry powder formulations described herein preferably have a specific mass median aerodynamic diameter (MMAD) when provided in aerosol form. The microparticles may also have a specific optical volume mean diameter (oVMD). The oVMD may also be referred to as the particle size distribution (PSD or pPSD).

[0070] As used herein, "mass median aerodynamic diameter" or "MMAD" is a measure of the aerodynamic size of dispersed fine particles. Aerodynamic diameter is used to describe an aerosolized powder in terms of its settling behavior and is the diameter of a unit density sphere that has the same settling velocity in air as the fine particle. Aerodynamic diameter encompasses the particle shape, density, and physical size of the fine particle. As used herein, MMAD refers to the midpoint or median of the aerodynamic particle size distribution of an aerosolized powder as determined by cascade impaction, unless otherwise indicated. Preferably, the microparticles of the dry powder formulations provided herein have a mass median aerodynamic diameter (MMAD) of at least 1 μm or more, more preferably about 1 μm to about 10 μm, about 2 μm to about 8 μm, about 2 μm to about 7 μm, about 2 μm to about 6 μm, about 2 μm to about 5 μm, about 2 μm to about 4 μm, about 3 μm to about 7 μm, about 4 μm to about 7 μm, about 3 μm to about 6 μm, or about 2 μm, about 3 μm, about 4 μm, about 5 μm, about 6 μm, or about 7 μm.

[0071] Preferably, the fine particle fraction (the fraction of particles emitted from an inhalation device having an aerodynamic diameter of less than 5 μm for the dry powder formulations described herein) is 50% or greater, more preferably 60% or greater. This fine particle fraction (FPF) can contribute to low device retention, where less than 20% (preferably less than 15%, less than 10%, or less than 5%) of the dry powder formulation remains in the device after delivery to a patient.

[0072] In additional embodiments, the microparticles preferably have an equivalent optical volume mean diameter of about 0.5 μm to about 7 μm. Equivalent optical volume mean diameter (oVMD) refers to the average diameter of a sphere that best approximates a particular optical interaction of the microparticle with light, when measured using an appropriate optical technique, where half of the microparticles are best approximated by an equivalent sphere that is smaller, and half of the microparticles are best approximated by an equivalent sphere that is larger than the average. In exemplary embodiments, the microparticles have an equivalent optical volume mean diameter (oVMD) of about 0.5 μm to about 6 μm, or about 1 μm to about 5 μm, or about 1 μm to about 4 μm, or about 2 μm to about 4.5 μm, or about 2.5 μm to about 4 μm, or about 2 μm to about 4 μm, or about 2 μm to about 3 μm, or about 2 μm to about 3.5 μm, or about 1 μm, about 1.5 μm, about 2 μm, about 2.5 μm, about 3 μm, about 3.5 μm, about 4 μm, about 4.5 μm, or about 5 μm.

[0073] As described herein, a high compacted bulk density allows for the delivery of a larger amount of active agent using the same delivery volume. Certain biological agents may require a delivery payload of 50 mg / dose or more for effective treatment. As illustrated in Figure 2B, the combination of leucine and trileucine can result in a dry powder formulation with a higher bulk density, and therefore a substantially smaller volume for the same fill weight.

[0074] Exemplary platform formulations shown in Figure 2B are listed below: LTC indicates a formulation that does not contain trileucine (TLeu) but does contain leucine, trehalose, and a citrate buffer; TTC indicates a formulation that does not contain leucine (Leu) but does contain trileucine, trehalose, and a citrate buffer; TLTC indicates the inclusion of both leucine and trileucine, as well as trehalose and a citrate buffer. Cit refers to a citrate buffer. Tre refers to trehalose.

[0075] [Table 1]

[0076] The capsules for each formulation (size 3 capsules) are shown in Figure 2B with their respective fill weights. As shown, in the case of the TLTC formulation, the combination of trileucine and leucine allows for the filling of 100 mg of dry powder formulation into capsules while still maintaining some residual space within the capsule. The other formulations were unable to fill more than approximately 70-80 mg. This represents a dramatic improvement in the use of leucine and trileucine in preparing formulations with high compacted bulk densities, allowing for higher fill weights.

[0077] As described herein, the use of leucine and trileucine in the present dry powder formulations also results in microparticles with a desired size (MMAD), as well as a desired specific surface area (SSA) and roughness, resulting in microparticles that can flow adequately and be delivered to the lungs using a variety of inhalation platforms.

[0078] The specific surface area (SSA) of a particulate is defined as the total surface area of ​​the particulate per unit mass (preferably in units m 2 / g). Methods for measuring SSA are known in the art, including, for example, Brunauer-Emmett-Teller (BET) measurements, which use an evaluation of the specific surface area of ​​a material by nitrogen adsorption measured as a function of relative pressure. This surface area is determined by calculating the amount of adsorbed gas corresponding to a monolayer on the surface of the microparticle. This technique measures the external area and an evaluation of any pore area to determine the total specific surface area. Instruments for measuring BET are known in the art.

[0079] In embodiments, the specific surface area (SSA) of the microparticles of the dry powder formulation is about 3 m 2 / g~about 8m 2 In a preferred embodiment, the SSA of the plurality of particulates is about 3.5 m / g. 2 / g~7.5m 2 / g, or approximately 4m 2 / g~7m 2 / g, or approximately 4.5m 2 / g~7m 2 / g, or approximately 5m 2 / g~7m 2 / g, or approximately 4.5m 2 / g~6m 2 / g, or approximately 5m 2 / g~6m 2 / g, or approximately 4m 2 / g, approx. 4.5m 2 / g, approx. 5m 2 / g, approx. 5.5m 2 / g, approx. 6m 2 / g, approx. 6.5m 2 / g, or approximately 7m 2 / g.

[0080] Figure 3 shows the specific surface area (m 2 3 shows the SSA content of the microparticles at approximately 500 wt% of the trileucine in the formulation. Each column in FIG. 3 represents a different amount of trileucine in the formulation. Within each column, the amount of leucine increases from about 1% to about 20%. The inserted photomicrographs show the physical appearance of microparticles with low SSA (bottom left) and higher SSA (top right). As shown, at the lower wt% trileucine, the SSA is approximately 500 wt%. 2 / g, but increases with increasing leucine. Above about 1% trileucine, SSA is 3.0m 2 / g and increases as the proportion of leucine increases. 2 / g and is over 7.0m 2 SSA values ​​approaching 1 / g are achieved with trileucine levels above about 4%. 2 The desired range of specific surface area / g can be easily achieved by using amounts of about 1-6% trileucine and about 1-20% leucine. As shown, by using an amount of trileucine less than about 6%, the amount of leucine can be kept below 10%, or even below 5%, and still maintain microparticles with the desired SSA and surface roughness. The photomicrograph on the top left shows the shape of microparticles of the dry powder formulation described herein, exhibiting the desired size, specific surface area, and surface roughness.

[0081] In certain embodiments, the dry powder formulation has a compressed bulk density of about 0.4-1.0 g / cm 3 Preferably, the compressed bulk density of the dry powder formulation is about 0.5 to 0.8 g / cm 3 In embodiments, the compressed bulk density of the dry powder formulations described herein is about 0.4 to 0.9 gm / cm 3 , about 0.4~0.8gm / cm 3 , about 0.5~0.8gm / cm 3 , about 0.6~0.8gm / cm 3 , or about 0.4 gm / cm 3 , about 0.5gm / cm 3 , about 0.6gm / cm 3 , about 0.7gm / cm 3 , or about 0.8 gm / cm 3 In certain embodiments, the compressed bulk density of the dry powder formulations described herein is about 0.4 gm / cm 3 ~about 0.9gm / cm 3 In certain embodiments, the compressed bulk density of the dry powder formulations described herein is about 0.5 gm / cm 3 ~about 0.8gm / cm 3 is.

[0082] The dry powder formulations preferably include a glass stabilizer (e.g., an amorphous sugar or a buffer, or both an amorphous sugar and a buffer) as described herein. Exemplary amorphous sugars include those described herein, such as trehalose, sucrose, raffinose, inulin, dextran, and cyclodextrin. Preferably, the amorphous sugar is present at about 30% to about 70%, and in embodiments, is trehalose (preferably present at about 35% to 60% or 35% to 55%).

[0083] Exemplary buffers for use in the dry powder formulations are described herein and include citrate buffers, phosphate buffers, and tartrate buffers. Preferably, the buffer is present at about 1% to about 10%, and in embodiments, is a citrate buffer. In certain embodiments, the pH of the citrate buffer is about pH 5.5 to about pH 6.5, e.g., about pH 5.5, about pH 5.6, about pH 5.7, about pH 5.8, about pH 5.9, about pH 6.0, about pH 6.1, about pH 6.2, about pH 6.3, about pH 6.4, or about pH 6.5. In certain embodiments, the pH of the citrate buffer is about pH 6.4.

[0084] In certain embodiments, the dry powder formulations described herein contain a surfactant. As defined herein, "surfactant" refers to a molecule or compound that reduces particle clumping and adhesion to the surfaces of capsules, container walls, or valve components of inhalable delivery devices. Surfactants have also been shown to reduce the formation of subvisible particles (SVPs) upon reconstitution of the formulation. Removing or reducing SVP formation simplifies analytical characterization of the formulation by removing the burden of tracking SVP formation during manufacturing. Analytical characterization of SVPs may involve the development of orthogonal techniques to identify and quantify SVPs for quality control purposes. Therefore, removing SVPs or reducing them to acceptable levels eliminates this characterization step from the manufacturing process, streamlining manufacturing. Additionally, because the kinetics of drug release from SVPs are unknown, removal of SVPs may result in a more predictable dose range. Furthermore, removal of SVPs may increase the amount of active agent available to participate in pharmacological activity after reconstitution, which may mean that not only may higher delivered doses be achieved, but also more accurate delivered dose predictions may be calculated. Higher delivered doses may also benefit the patient, for example, by potentially reducing the number or frequency of doses that must be delivered to elicit pharmacological benefit.

[0085] "Sub-visible to the naked eye" ("SVP") refers to particles that are invisible to the naked eye, between about 1 μm and about 200 μm. The presence of sub-visible particles can be inferred upon reconstitution of a dry powder formulation by the cloudiness of the reconstituted liquid. Accurate determination of the presence of SVPs can be confirmed using techniques such as microflow imaging. Microflow imaging (i.e., MFI) combines microfluidic flow microscopy with high-resolution imaging particle analysis to quantify SVP counts. MFI can bin the number of SVPs across a range of particle sizes, e.g., by binning particle counts in size ranges of about 1 to about 200 μm, about 2 μm to about 200 μm, about 5 μm to about 200 μm, about 10 μm to about 200 μm, and about 25 μm to about 200 μm. The examples show that the inclusion of a surfactant reduces the presence of SVPs in each particle size range compared to a control formulation without the surfactant present (see Figure 10A). Thus, in certain embodiments, the dry powder formulations disclosed herein contain a surfactant, and upon reconstitution, the formulation has a reduced number of subvisible particles, hi some embodiments, the number of subvisible particles is reduced compared to an equivalent formulation that does not contain a surfactant.

[0086] In certain embodiments, the number of SVPs having a size of about 25 μm to about 200 μm is reduced to less than 30,000 particles per ml, e.g., 25,000 particles per ml, 20,000 particles per ml, 15,000 particles per ml, 10,000 particles per ml, or 5,000 particles per ml. In certain embodiments, the number of SVPs having a size of about 25 μm to about 200 μm is reduced to less than 1,000 particles per ml. In certain embodiments, the number of SVPs having a size of about 25 μm to about 200 μm is reduced to less than 1,000 particles per ml. In certain embodiments, the number of SVPs having a size of about 25 μm to about 200 μm is reduced to less than 100 particles per ml.

[0087] In certain embodiments, the number of SVPs having a size of about 10 μm to about 200 μm is reduced to less than 100,000 particles per ml, e.g., 90,000 particles per ml, 80,000 particles per ml, 70,000 particles per ml, 60,000 particles per ml, 50,000 particles per ml, 40,000 particles per ml, or 30,000 particles per ml. In certain embodiments, the number of SVPs having a size of about 10 μm to about 200 μm is reduced to less than 10,000 particles per ml. In certain embodiments, the number of SVPs having a size of about 10 μm to about 200 μm is reduced to less than 1,000 particles per ml. In certain embodiments, the number of SVPs between about 10 μm and about 200 μm in size is reduced to less than 100 particles per ml.

[0088] In certain embodiments, the number of SVPs having a size of about 5 μm to about 200 μm is reduced to less than 200,000 particles per ml, e.g., 180,000 particles per ml, 170,000 particles per ml, 160,000 particles per ml, 150,000 particles per ml, or 140,000 particles per ml. In certain embodiments, the number of SVPs having a size of about 5 μm to about 200 μm is reduced to less than 50,000 particles per ml. In certain embodiments, the number of SVPs having a size of about 5 μm to about 200 μm is reduced to less than 10,000 particles per ml. In certain embodiments, the number of SVPs having a size of about 5 μm to about 200 μm is reduced to less than 2,000 particles per ml.

[0089] In certain embodiments, the number of SVPs having a size of about 2 μm to about 200 μm is 1×10 per ml. 6 The concentration of particles has been reduced to less than 0.8 × 10 per ml. 6 0.7 x 10 particles per ml 6 0.6 x 10 particles per ml 6 particles, or 0.5 x 10 per ml6 In certain embodiments, the number of SVPs having a size of about 2 μm to about 200 μm is reduced to less than 100,000 particles per ml. In certain embodiments, the number of SVPs having a size of about 2 μm to about 200 μm is reduced to less than 50,000 particles per ml. In certain embodiments, the number of SVPs having a size of about 2 μm to about 200 μm is reduced to less than 10,000 particles per ml.

[0090] In certain embodiments, the number of SVPs having a size of about 1 μm to about 200 μm is 2×10 per ml. 6 Reduced to less than 1.8 x 10 particles per ml 6 1.7 x 10 particles per ml 6 1.6 x 10 particles per ml 6 particles, or 1.5 x 10 per ml 6 In certain embodiments, the number of SVPs between about 1 μm and about 200 μm in size is reduced to less than 200,000 particles per ml. In certain embodiments, the number of SVPs between about 1 μm and about 200 μm in size is reduced to less than 150,000 particles per ml.

[0091] In certain embodiments, the number of SVPs between about 25 μm and about 200 μm in size is reduced by more than 2-fold upon reconstitution compared to the reference control, e.g., by more than 3-fold, 4-fold, 5-fold, 6-fold, 7-fold, 8-fold, or 9-fold. In certain embodiments, the number of SVPs between about 25 μm and about 200 μm in size is reduced by more than 10-fold upon reconstitution compared to the reference control.

[0092] In certain embodiments, the number of SVPs between about 10 μm and about 200 μm in size is reduced by more than 2-fold upon reconstitution compared to the reference control, e.g., by more than 3-fold, 4-fold, 5-fold, 6-fold, 7-fold, 8-fold, or 9-fold. In certain embodiments, the number of SVPs between about 10 μm and about 200 μm in size is reduced by more than 10-fold upon reconstitution compared to the reference control.

[0093] In certain embodiments, the number of SVPs between about 5 μm and about 200 μm in size is reduced by more than 2-fold upon reconstitution compared to the reference control, e.g., by more than 3-fold, 4-fold, 5-fold, 6-fold, 7-fold, 8-fold, or 9-fold. In certain embodiments, the number of SVPs between about 5 μm and about 200 μm in size is reduced by more than 10-fold upon reconstitution compared to the reference control.

[0094] In certain embodiments, the number of SVPs between about 2 μm and about 200 μm in size is reduced by more than 2-fold upon reconstitution compared to the reference control, e.g., by more than 3-fold, 4-fold, 5-fold, 6-fold, 7-fold, 8-fold, or 9-fold. In certain embodiments, the number of SVPs between about 2 μm and about 200 μm in size is reduced by more than 10-fold upon reconstitution compared to the reference control. In certain embodiments, the number of SVPs between about 2 μm and about 200 μm in size is reduced by more than 100-fold upon reconstitution compared to the reference control.

[0095] In certain embodiments, the number of SVPs between about 1 μm and about 200 μm in size is reduced by more than 2-fold upon reconstitution compared to the reference control, e.g., by more than 3-fold, 4-fold, 5-fold, 6-fold, 7-fold, 8-fold, or 9-fold. In certain embodiments, the number of SVPs between about 1 μm and about 200 μm in size is reduced by more than 10-fold upon reconstitution compared to the reference control.

[0096] In certain embodiments, the reference control is an equivalent formulation lacking surfactant. In some embodiments, the formulation is reconstituted with water. In some embodiments, the formulation is reconstituted to an active agent concentration of 30 mg / ml. In some embodiments, the formulation is reconstituted to an active agent concentration of 2.5 mg / ml. In some embodiments, the number of SVPs is determined by microflow imaging (MFI).

[0097] Exemplary surfactants suitable for use in the dry powder formulations described herein include, but are not limited to, polysorbate-20 (PS-20), polysorbate-40 (PS-40), polysorbate-60 (PS-60), polysorbate-80 (PS-80), and poloxamer-188. In certain embodiments, the formulations described herein include PS-80, preferably at a concentration ranging from about 0.27% to about 2.7% by weight, preferably from about 0.27% to about 1.33% by weight, and preferably from about 0.67% to about 1.33% by weight. In certain embodiments, the formulations include PS-80 at a concentration ranging from about 0.3% to about 3% by weight. In certain embodiments, the formulations include PS-80 at a concentration ranging from about 0.3% to about 2.5% by weight. In certain embodiments, the formulation contains PS-80 at a concentration ranging from about 0.5% to about 2.5% by weight. In certain embodiments, the formulation contains PS-80 at a concentration ranging from about 0.5% to about 2% by weight. In certain embodiments, the formulation contains PS-80 at a concentration ranging from about 0.5% to about 1.5% by weight.

[0098] In an exemplary embodiment, the formulation comprises PS-80 at a concentration ranging from about 0.67% to about 1.33%.

[0099] In exemplary embodiments, the formulation comprises PS-80 at a concentration of about 0.7% (w / w), about 0.8% (w / w), about 0.9% (w / w), about 1.0% (w / w), about 1.1% (w / w), about 1.2% (w / w), or about 1.3% (w / w). In some embodiments, the formulation comprises PS-80 at a concentration of about 1.1% (w / w).

[0100] In exemplary embodiments, the composition comprises PS-80 at a concentration of 0.7%±0.35 (w / w), about 0.8%±0.4 (w / w), about 0.9%±0.45 (w / w), about 1.0%±0.5 (w / w), about 1.1%±0.55 (w / w), about 1.2%±0.6 (w / w), or about 1.3%±0.65 (w / w). In some embodiments, the formulation comprises PS-80 at a concentration of 1.1%±0.55 (w / w).

[0101] In exemplary embodiments, the composition comprises PS-80 at a concentration of 0.7%±0.35 (w / w), about 0.8%±0.4 (w / w), about 0.9%±0.45 (w / w), about 1.0%±0.5 (w / w), about 1.1%±0.55 (w / w), about 1.2%±0.6 (w / w), about 1.3%±0.65 (w / w), about 1.4%±0.7 (w / w), about 1.5%±0.75 (w / w), about 1.6%±0.8 (w / w), or about 1.7%±0.75 (w / w).

[0102] In certain embodiments, the formulations described herein include poloxamer-188, preferably at a concentration ranging from about 1% to about 10% by weight. In exemplary embodiments, the formulations include poloxamer-188 (P188) at a concentration ranging from about 0.67% to about 2.67%. In certain embodiments, the formulations include P188 at a concentration ranging from about 0.3% to about 3% by weight. In certain embodiments, the formulations include P188 at a concentration ranging from about 0.3% to about 2.5% by weight. In certain embodiments, the formulations include P188 at a concentration ranging from about 0.5% to about 2.5% by weight. In certain embodiments, the formulations include P188 at a concentration ranging from about 0.5% to about 2% by weight. In certain embodiments, the formulations include P188 at a concentration ranging from about 0.5% to about 1.5% by weight.

[0103] In an exemplary embodiment, the formulation comprises P188 at a concentration ranging from about 0.67% to about 1.67%.

[0104] In exemplary embodiments, the formulation comprises P188 at a concentration of about 0.7% (w / w), about 0.8% (w / w), about 0.9% (w / w), about 1.0% (w / w), about 1.1% (w / w), about 1.2% (w / w), about 1.3% (w / w), about 1.4% (w / w), about 1.5% (w / w), about 1.6% (w / w), or about 1.7% (w / w).

[0105] In an exemplary embodiment, the dry powder formulation comprises about 39% trehalose, about 10.5% leucine, about 2% trileucine, about 8.5% citrate buffer, and an active agent.

[0106] Exemplary active agents are described throughout the specification and include small molecules and biologics, such as antibodies and antigen-binding fragments thereof.

[0107] Suitable sizes of the microparticles of the dry powder formulations are described herein, and in embodiments, the plurality of microparticles have a mass median aerodynamic diameter (MMAD) of about 2 μm to about 4 μm when provided in aerosol form. Suitable specific surface areas (SSA) of the microparticles are described herein, for example, about 4 to 7 m. 2 / g. Preferably, the microparticles have an equivalent optical volume mean diameter (oVMD) of about 1 μm to about 5 μm.

[0108] In a further embodiment, provided herein is a method for preparing a dry powder formulation. In an embodiment, the method includes preparing a liquid feedstock preferably containing leucine, about 0.1 mg / mL to about 6 mg / mL trileucine, an active agent, and preferably further containing a glass stabilizer. The glass stabilizer described herein may be omitted from the dry powder formulation if desired. The liquid feedstock may also include a surfactant. The liquid feedstock is prepared by combining these components in a liquid solvent to create a feedstock in which each of these components is dissolved. Heating may be applied, if desired or necessary, to increase the solubility of the various components forming the liquid feedstock. Exemplary liquid solvents include water, e.g., deionized water, and dilute solutions of alcohols in water. In an embodiment, the active agent is preferably added to the liquid feedstock after the addition and dissolution of the remaining ingredients of the feedstock.

[0109] In a preferred embodiment of this preparation method, the leucine and trileucine are present in the liquid feedstock in a concentration ratio of leucine:trileucine of about 0.1:1 to about 30:1. When preparing the liquid feedstock as described herein, the leucine and trileucine are provided in mg / mL amounts. Thus, in such embodiments, a concentration ratio of leucine:trileucine of about 0.1:1 to about 30:1 in a given volume of liquid feedstock corresponds to the ratio of leucine:trileucine by weight in the liquid feedstock. In further embodiments, the leucine and trileucine are present in the liquid feedstock in a concentration ratio of leucine:trileucine of about 0.1:1 to about 25:1, about 0.5:1 to about 20:1, about 1:1 to about 20:1, about 1:1 to about 15:1, about 1:1 to about 12:1, about 1:1 to about 10:1, about 1:1 to about 7:1, about 1:1 to about 6:1, or about 1:1, about 2:1, about 3:1, about 4:1, about 5:1, about 5:1:1, about 5.2:1, about 5.25:1, about 5.3:1, about 5.4:1, about 5.5:1, about 5.75:1, or about 6:1.

[0110] The liquid feedstock may then be atomized. In certain embodiments, the liquid feedstock is filtered before atomization. In certain embodiments, the liquid feedstock is filtered through a 0.22 micron filter. In certain embodiments, the liquid feedstock containing leucine and trileucine is filtered before the addition of the activator. In certain embodiments, the liquid feedstock is filtered after the addition of the activator before atomization. Atomization refers to converting the liquid feedstock into fine droplets, preferably using pressurized gas (e.g., CO or an inert gas). Exemplary devices for producing atomized liquid feedstocks are known in the art and include the use of various spray nozzles having the desired size and flow characteristics. Exemplary parameters for atomization include an outlet temperature of about 50°C to 90°C (preferably about 60°C to 80°C or about 70°C); a feed rate of about 8 to 15 ml / min (preferably about 9 to 14 ml / min, about 10 to 13 ml / min, or about 12 ml / min); an atomizer gas feed rate of about 9 to 15 kg / hour (hr or h) (preferably about 10 to 14 kg / hr, about 12 to 14 kg / hr, or about 13 kg / hr); and a drying gas feed rate of about 60 to 100 kg / hr (preferably about 60 to 90 kg / hr, about 70 to 90 kg / hr, or about 80 kg / hr).

[0111] The atomized liquid feedstock may then be dried, preferably under heat and in combination with flowing air to aid drying. This drying results in a plurality of microparticles. Drying temperatures are typically in the range of about 50°C to 100°C, or about 60°C to 100°C, or about 70°C to 90°C, and air flow rates are typically in the range of about 10 to 40 m / s. 3 / It can be on the order of hours.

[0112] Exemplary glass stabilizers (e.g., amorphous sugars and buffers) are described herein, as are suitable amounts of the glass stabilizers. Suitable amounts of leucine and trileucine are also described throughout. The final dry powder formulation should contain the recited amounts of leucine and trileucine (as well as other ingredients), so such amounts are also used in the liquid ingredients. The drying process after spraying results in the removal of any liquid solvent, so that the total original dry weight of the ingredients corresponds to the final dry weight of the ingredients in the dry powder formulation. Exemplary active agents are also described herein.

[0113] The methods for preparing dry powder formulations described herein preferably result in microparticles having the desired physical properties noted (e.g., desired compacted bulk density, specific surface area, and size). Exemplary sizes, as well as exemplary SSAs, are described herein and are approximately 10 m 2 / g (preferably about 4 to 7 m 2 Preferably, the method produces a powder having an equivalent optical volume mean diameter (oVMD) of about 1 μm to about 5 μm; a mass median aerodynamic diameter (MMAD) of about 2 μm to about 4 μm when provided in aerosol form; and a compressed bulk density of about 0.4 g / cm, as described herein. 3 ~0.8g / cm 3 A plurality of microparticles having the formula:

[0114] A further advantage of the methods for preparing dry powder formulations described herein relates to the high throughput of the process. For example, when the spray feed rate was set at 20 ml / min, the following throughput (grams / hour) was determined:

[0115] [Table 2]

[0116] As noted, using only trileucine in the feedstock at a maximum trileucine concentration of 5 mg / mL resulted in a maximum solids loading of 25 mg / mL (related to maximum solubility). This resulted in a throughput of 30 g / hour. With only 60% leucine at a maximum leucine concentration of 20 mg / mL, a maximum solids loading of 33 mg / mL and a throughput of 40 g / hour were achieved. Further results for the use of leucine and trileucine alone are also shown. In contrast, for the three feedstocks tested containing both leucine and trileucine, a maximum solids loading of 250 mg / mL and a throughput of 300 g / hour were achieved using only 8% leucine and 2% trileucine. This was an unexpected and unpredictable finding of the advantages of the methods and formulations disclosed herein, in that dispersible particles can be obtained using relatively small amounts of leucine and trileucine, yet still allowing for high throughput. Such high throughput has a significant impact on the ability to scale up the production of the dry powder formulations described herein, where large amounts of formulation are required.

[0117] The methods and formulations described herein enable the manufacture of capsules, blister packs, and other containers suitable for dry powder formulations. Such containers may be manufactured with 10-200 mg of dry powder, preferably 10-100 mg, or 25-75 mg, or 50 mg of dry powder formulation. Such containers may preferably deliver 0.1-10 mg of dry powder formulation to a patient's lungs.

[0118] In some embodiments, use of the methods described herein results in dry powder formulations that may reduce the total number of capsules required for use in an inhalation device. For example, the amount required to deliver 50-100 mg of active agent may be reduced from two larger size 00 capsules to a single size 3 capsule.

[0119] The methods described herein also provide a mechanism for increasing the compacted bulk density and specific surface area of ​​dry powder formulations containing a plurality of microparticles. The incorporation of leucine and trileucine into the dry powder formulations increases the compacted bulk density and specific surface area to approximately 0.4-1.0 g / cm. 3 (Preferably about 0.5 to 0.8 g / cm 3 ) can easily achieve a compacted bulk density of about 5 to 10 m 2 / g (preferably about 5m 2 / g ~ approx. 7m 2 / g) can also be achieved. In additional embodiments, the size of the microparticles can be formed within the ranges described herein (e.g., microparticles having a mass median aerodynamic diameter (MMAD) of about 2 μm to about 4 μm when provided in aerosol form).

[0120] In an embodiment, the compressed bulk density is about 0.4 to about 1.0 g / cm 3 The present invention provides a method for preparing a dry powder formulation comprising a plurality of microparticles, the method comprising incorporating into the dry powder formulation leucine and trileucine in a ratio of leucine:trileucine of about 0.1:1 to about 30:1 by weight.

[0121] In a further embodiment, the specific surface area is about 5 to about 10 m 2 A method is provided for preparing a dry powder formulation comprising a plurality of particulates, where the particulates are soluble in water / g, the method comprising incorporating into the dry powder formulation leucine and trileucine in a ratio of leucine:trileucine of about 0.1:1 to about 30:1 by weight.

[0122] In a further embodiment, a method is provided for preparing a dry powder formulation comprising a plurality of microparticles, the microparticles having a mass median aerodynamic diameter (MMAD) of from about 2 μm to about 4 μm when provided in aerosol form, the method comprising incorporating leucine and trileucine into the dry powder formulation in a ratio of leucine to trileucine of from about 0.1:1 to about 30:1 by weight.

[0123] Suitable amounts of leucine and trileucine that can be utilized in methods to increase compacted bulk density, achieve specific contact area of ​​a dry powder formulation, and / or achieve a particular mass median aerodynamic diameter are described herein and include incorporating about 5% to about 15% leucine and about 1% to about 5% trileucine, preferably about 8% to about 11% leucine and about 2% to about 4% trileucine. In embodiments, the method includes incorporating about 10.5% leucine and about 2% trileucine.

[0124] Also provided herein are methods of delivering the dry powder formulations described herein to the lungs of a mammalian patient. Preferably, such methods involve inhalation of the dry powder formulation in aerosol form. Mammalian patients to which the dry powder formulations can be administered include humans and other mammals, such as dogs, cats, sheep, pigs, cattle, primates, etc.

[0125] Methods of producing aerosol forms of dry powder preparations are known in the art and include, for example, the use of an inhaler device such as a dry powder inhaler (DPI) (e.g., the Monodose RS01 DPI from PLASTIAPE, Osnago, Italy). The dry powder formulations described herein can be dispensed into a gas stream by either a passive or active inhalation device and can remain suspended in the gas for a sufficient time for at least a portion of the microparticles to be inhaled by the patient so that some of the microparticles reach the lungs.

[0126] Also provided herein is a method of treating a medical condition in a mammalian patient, comprising administering to the patient by inhalation (e.g., by a dry powder inhaler) a dry powder formulation described herein.

[0127] Medical conditions that may be treated using the methods described herein include those affecting the nervous system, endocrine system, muscular system, cardiovascular system, digestive system, respiratory system (especially the lungs), hormonal system, immune system, reproductive system, etc.

[0128] Additional Exemplary Embodiments Embodiment 1 is a dry powder formulation comprising a plurality of microparticles, the microparticles comprising leucine, about 0.5% to about 10% by weight trileucine, and an active agent, wherein the leucine and trileucine are present in a ratio of leucine:trileucine of about 0.1:1 to about 30:1 by weight.

[0129] In embodiment 2, the dry powder formulation is the dry powder formulation of embodiment 1, wherein the compressed bulk density is about 0.4 to about 1.0 g / cm 3 .

[0130] Embodiment 3 is a dry powder formulation of embodiment 1 or 2, further comprising a glass stabilizer.

[0131] Embodiment 4 is the dry powder formulation of embodiment 3, wherein the glass stabilizer is an amorphous sugar or a buffering agent.

[0132] Embodiment 5 is the dry powder formulation of embodiment 3, wherein the glass stabilizer comprises an amorphous sugar and a buffering agent.

[0133] Embodiment 6 is the dry powder formulation of embodiment 4 or 5, wherein the amorphous sugar is selected from the group consisting of trehalose, sucrose, raffinose, inulin, dextran, mannitol, and cyclodextrin.

[0134] Embodiment 7 is the dry powder formulation of any one of Embodiments 4 to 6, wherein the buffer is selected from the group consisting of citrate buffer, phosphate buffer, histidine buffer, glycine buffer, acetate buffer, and tartrate buffer.

[0135] Embodiment 8 is the dry powder formulation of any one of Embodiments 4 to 7, wherein the amorphous sugar is present in an amount of about 30% to about 70% by weight.

[0136] Embodiment 9 is the dry powder formulation of any one of Embodiments 4 to 8, wherein the amorphous sugar is trehalose.

[0137] Embodiment 10 is the dry powder formulation of embodiment 9, wherein the trehalose is present in about 30% to 65% by weight.

[0138] Embodiment 11 is the dry powder formulation of any one of Embodiments 4 to 10, wherein the buffering agent is present at about 1% to about 10% by weight.

[0139] Embodiment 12 is a dry powder formulation of any one of embodiments 1 to 11, wherein the active agent is a small molecule.

[0140] Embodiment 13 is a dry powder formulation of any one of embodiments 1 to 12, wherein the active agent is a biologic.

[0141] Embodiment 14 is the dry powder formulation of embodiment 13, wherein the biologic is an antibody or an antigen-binding fragment thereof.

[0142] Embodiment 15 is a dry powder formulation of any one of embodiments 1-14, wherein the ratio of leucine:trileucine is from about 1:1 to about 12:1 by weight.

[0143] Embodiment 16 is a powder dry formulation of any one of embodiments 1 to 15, wherein the ratio of leucine:trileucine is from about 1:1 to about 7:1 by weight.

[0144] Embodiment 17 is a dry powder formulation of any one of embodiments 1-16, wherein the ratio of leucine:trileucine is about 5.25:1 by weight.

[0145] Embodiment 18 is a dry powder formulation of any one of Embodiments 1-17 comprising about 1% to about 10% (optionally about 5%) trileucine by weight.

[0146] Embodiment 19 is the dry powder formulation of any one of Embodiments 1 to 18, comprising about 8% to about 11% by weight leucine and about 2% to about 4% by weight trileucine.

[0147] Embodiment 20 is a dry powder formulation of any one of Embodiments 1-19, comprising about 10.5% by weight leucine and about 2% by weight trileucine.

[0148] Embodiment 21 is the dry powder formulation of any one of embodiments 1 to 20, further comprising a surfactant, optionally selected from polysorbate-20 (PS-20), polysorbate-40 (PS-40), polysorbate-60 (PS-60), polysorbate-80 (PS-80), and poloxamer-188.

[0149] Embodiment 22 is the dry powder formulation of embodiment 21, wherein the surfactant is PS-80, and optionally, the PS-80 is present at a concentration ranging from about 0.27% to about 2.7% by weight.

[0150] Embodiment 23 is a dry powder formulation of embodiment 22, wherein PS-80 is present in a concentration of about 1.1% by weight.

[0151] Embodiment 23 is a dry powder formulation of embodiment 21, wherein the surfactant is poloxamer-188, and optionally, the poloxamer-188 is present in a concentration ranging from about 1% to about 10% by weight.

[0152] Embodiment 25 is a dry powder formulation of claim 23, wherein poloxamer-188 is present in a concentration ranging from about 0.67% to about 1.0% by weight.

[0153] Embodiment 26 is the dry powder formulation of any one of embodiments 1 to 25, wherein the plurality of microparticles has an equivalent optical volume mean diameter (oVMD) of about 1 μm to about 5 μm.

[0154] Embodiment 27 is a dry powder formulation of any one of embodiments 1 to 26, wherein the plurality of microparticles, when provided in aerosol form, has a mass median aerodynamic diameter (MMAD) of about 2 μm to about 4 μm.

[0155] Embodiment 28 is a dry powder formulation of any one of Embodiments 2 to 27, having a compressed bulk density of about 0.5 g / cm 3 to about 0.8 g / cm 3 .

[0156] Embodiment 29 is a dry powder formulation of any one of embodiments 1-28, comprising about 39% trehalose, about 10.5% leucine, about 2% trileucine, and about 8.5% citrate buffer.

[0157] Embodiment 30 is the dry powder formulation of any one of embodiments 1 to 29, wherein the plurality of microparticles has a specific surface area of ​​less than about 10 m2 / g.

[0158] Embodiment 31 is the dry powder formulation of embodiment 30, wherein the plurality of microparticles has a specific surface area of ​​about 4 mg2 / g to about 7 m2 / g.

[0159] Embodiment 32 is a dry powder formulation comprising a plurality of microparticles, the microparticles comprising about 10.5% by weight leucine, about 2% by weight trileucine, about 8.5% by weight citrate buffer, about 1% to about 40% by weight active agent, about 1.07% by weight polysorbate-80, and trehalose in amounts by weight that total 100%.

[0160] Embodiment 33 is a dry powder formulation of embodiment 32, wherein the active agent is a biologic.

[0161] Embodiment 34 is the dry powder formulation of embodiment 33, wherein the biologic is an antibody or an antigen-binding fragment thereof.

[0162] Embodiment 35 is a method of preparing a dry powder formulation, comprising: i. preparing a liquid feedstock comprising leucine, about 0.1 mg / mL to about 6 mg / mL trileucine, an active agent, and a liquid solvent, wherein the leucine and trileucine are present in a concentration ratio of about 0.1:1 to about 30:1 leucine:trileucine; ii. spraying the liquid feedstock; and iii. drying the spray-dried liquid feedstock to form a plurality of microparticles.

[0163] Embodiment 36 is the method of embodiment 35, wherein the liquid ingredient further comprises a glass stabilizer.

[0164] Embodiment 37 is the method of embodiment 36, wherein the glass stabilizer is an amorphous sugar or a buffering agent.

[0165] Embodiment 38 is the method of embodiment 37, wherein the glass stabilizer comprises an amorphous sugar and a buffering agent.

[0166] Embodiment 39 is the method of embodiment 37 or 38, wherein the amorphous sugar is selected from the group consisting of trehalose, sucrose, raffinose, inulin, dextran, mannitol, and cyclodextrin.

[0167] Embodiment 40 is the method of any one of embodiments 37 to 39, wherein the buffer is selected from the group consisting of citrate buffer, phosphate buffer, histidine buffer, glycine buffer, acetate buffer, and tartrate buffer.

[0168] Embodiment 41 is the method of embodiment 39 or 40, wherein the amorphous sugar is present at about 30% to about 70%.

[0169] Embodiment 42 is the method of any one of embodiments 39-41, wherein the amorphous sugar is trehalose.

[0170] Embodiment 43 is the method of embodiment 42, wherein the trehalose is present at about 30% to about 65%.

[0171] Embodiment 44 is the method of any one of embodiments 37 to 43, wherein the buffering agent is present at about 1% to about 10%.

[0172] Embodiment 45 is the method of any one of embodiments 35 to 44, wherein the active agent is a small molecule.

[0173] Embodiment 46 is the method of any one of embodiments 35-44, wherein the active agent is a biologic.

[0174] Embodiment 47 is the method of embodiment 46, wherein the biologic is an antibody or an antigen-binding fragment thereof.

[0175] Embodiment 48 is the method of any one of embodiments 35 to 47, wherein step (a) further comprises adding a surfactant to the liquid feedstock, wherein the surfactant is optionally selected from polysorbate-20 (PS-20), polysorbate-40 (PS-40), polysorbate-60 (PS-60), polysorbate-80 (PS-80), and poloxamer-188.

[0176] Embodiment 49 is the method of embodiment 48, wherein the surfactant is PS-80, and optionally the PS-80 is present in the liquid feed at a concentration ranging from about 0.02% to about 0.2% by weight.

[0177] Embodiment 50 is the method of embodiment 48, wherein the surfactant is poloxamer-188, and the poloxamer-188 is present in the liquid feed at a concentration ranging from about 0.75% to about 7.5% by weight.

[0178] Embodiment 51 is the method of any one of embodiments 35 to 450, wherein the concentration ratio of leucine:trileucine in the feedstock is from about 1:1 to about 12:1.

[0179] Embodiment 52 is the method of any one of embodiments 35 to 50, wherein the concentration ratio of leucine:trileucine in the feedstock is from about 1:1 to about 7:1.

[0180] Embodiment 53 is the method of any one of embodiments 35 to 50, wherein the concentration ratio of leucine:trileucine in the feedstock is about 5.25:1.

[0181] Embodiment 54 is the method of embodiment 53, wherein the feedstock comprises about 1 mg / mL to about 1.7 mg / mL trileucine.

[0182] Embodiment 55 is the method of any one of embodiments 35 to 54, wherein the feedstock comprises about 1% to about 5% trileucine.

[0183] Embodiment 56 is the method of any one of embodiments 35 to 55, wherein the feedstock comprises about 8% to about 11% leucine and about 2% to about 4% trileucine.

[0184] Embodiment 57 is the method of any one of embodiments 35-56, wherein the feedstock comprises about 10.5% leucine and about 2% trileucine.

[0185] Embodiment 58 is the method of any one of embodiments 35 to 57, wherein the plurality of microparticles has an equivalent optical volume mean diameter (oVMD) of about 1 μm to about 5 μm.

[0186] Embodiment 59 is the method of any one of embodiments 35 to 58, wherein the plurality of microparticles, when provided in aerosol form, have a mass median aerodynamic diameter (MMAD) of about 2 μm to about 4 μm.

[0187] Embodiment 60 is the method of any one of embodiments 35 to 59, wherein the plurality of particulates has a compressed bulk density of about 0.4 g / cm 3 to 0.8 g / cm 3 .

[0188] Embodiment 61 is the method of any one of embodiments 35 to 60, wherein the plurality of particulates has a specific surface area of ​​less than about 10 m2 / g.

[0189] Embodiment 62 is the method of embodiment 61, wherein the plurality of particulates has a specific surface area of ​​about 4 m 2 / g to about 7 m 2 / g.

[0190] Embodiment 63 is the method of any one of embodiments 35 to 62, wherein the liquid solvent is water.

[0191] Embodiment 64 is a method of preparing a dry powder formulation comprising a plurality of particulates having a compressed bulk density of about 0.4 to about 1.0 g / cm, comprising incorporating leucine and trileucine into the dry powder formulation in a ratio of leucine:trileucine of about 0.1:1 to about 30:1 by weight.

[0192] Embodiment 65 is a method of preparing a dry powder formulation comprising a plurality of microparticles having a specific surface area of ​​about 5 to about 10 m / g, the method comprising incorporating leucine and trileucine into the dry powder formulation in a ratio of leucine:trileucine of about 0.1:1 to about 30:1 by weight.

[0193] Embodiment 66 is a method of preparing a dry powder formulation comprising a plurality of microparticles, the microparticles having a mass median aerodynamic diameter (MMAD) of from about 2 μm to about 4 μm when provided in aerosol form, the method comprising incorporating leucine and trileucine into the dry powder formulation in a ratio of leucine:trileucine of from about 0.1:1 to about 30:1 by weight.

[0194] Embodiment 67 is the method of any one of embodiments 64 to 66, comprising incorporating about 1% to about 5% by weight trileucine.

[0195] Embodiment 68 is the method of any one of embodiments 64 to 67, comprising incorporating about 8% to about 11% leucine and about 2% to about 4% trileucine.

[0196] Embodiment 69 is the method of any one of embodiments 64 to 68, comprising incorporating about 10.5% leucine and about 2% trileucine.

[0197] Embodiment 70 is the method of any one of embodiments 64-69, wherein the compressed bulk density is from about 0.4 g / cm 3 to about 0.8 g / cm 3 .

[0198] Embodiment 71 is the method of any one of embodiments 64 to 70, wherein the specific surface area is from about 5 m2 / g to about 7 m2 / g.

[0199] Embodiment 72 is a method of delivering a dry powder formulation to the lungs of a mammalian patient, comprising administering to the mammalian patient by inhalation the dry powder formulation of any one of embodiments 1 to 34 in aerosol form.

[0200] Embodiment 73 is the method of embodiment 72, wherein the dry powder formulation is administered by a dry powder inhaler (DPI).

[0201] Embodiment 74 is a method of treating a medical condition in a mammalian patient, comprising administering to the mammalian patient by inhalation a dry powder formulation according to any one of embodiments 1 to 34 in aerosol form.

[0202] Embodiment 75 is the method of embodiment 74, wherein the dry powder formulation is administered by a dry powder inhaler (DPI).

[0203] Embodiment 76 is a formulation according to any one of embodiments 1 to 34 for use in a method of treatment, wherein the formulation is administered by inhalation. [Example]

[0204] Example 1: Evaluation of the physical properties of spray-dried formulations containing leucine and trileucine The following method is used to evaluate the effect of the ratio of trileucine to leucine in the dry powder formulation on particle properties.

[0205] A total of 24 powders varying in wt% of trileucine, leucine, and trehalose (TLT) were spray dried in a pilot-scale spray dryer using identical process parameters at a 10% total feedstock solids concentration. Because the feedstock was prepared at a 10% (100 mg / mL) total solids concentration, all wt% values ​​in this study are also equivalent to concentration values ​​(mg / mL). The range of concentration values ​​for each particle additive is shown in Table 3.

[0206] [Table 3]

[0207] Each raw material (Table 4) was prepared by dissolving these additives in water. Once all additives were completely dissolved, the raw material was spray-dried using the following process parameters: outlet temperature, 70°C; raw material feed rate, 12 ml / min; nebulizer gas flow rate, 13 kg / hr; and drying gas flow rate, 80 kg / hr. These parameters were selected to achieve the particle and aerosol characteristics of a dry powder formulation for inhalation. Each of the 24 formulations was manufactured in 18 g batch sizes to obtain sufficient powder for characterization and product performance evaluation. Batches were randomly assigned and manufactured over two days.

[0208] [Table 4]

[0209] All formulations were tested for the following physical powder properties:

[0210] [Table 5]

[0211] The compressed bulk density (CBD) of the powders was measured using a GeoPyc® Model 1360 density analyzer (Micromeritics, Norcross, GA). Powder samples were prepared in a low humidity environment (<5% RH) and then transferred to the nitrogen gas-purged sample chamber of the density analyzer. The net weight of the powder sample was recorded, and then a compressive force of 12 N was applied to the sample by a plunger at a rate of 300 compaction strokes per second. The linear distance traveled by the plunger during each compaction stroke was converted to the volumetric displacement of the powder sample. The measurements from each compaction stroke were then averaged to produce a calculated bulk density value (g / cm). 3 (expressed as

[0212] The results show that the content of leucine and trileucine was found to significantly affect particle properties. Trileucine was identified as the primary factor with the greatest effect, and leucine was identified as the secondary factor with a similarly significant effect. The results are summarized in Table 6.

[0213] [Table 6]

[0214] Example 2 - Aerosol Performance Characteristics of Leucine / Trileucine Formulations The following examples evaluate the aerosol performance of formulations containing leucine and trileucine in a dry powder inhaler device. Twenty of the 24 formulations listed in Table 4 were tested for the aerosol performance outputs listed in Table 7. All product performance characterization was completed using a Monodose RS01 device with size 3 capsules. Next Generation Impactor (NGI) analysis was performed at a flow rate of 60 L / min.

[0215] USP <601> A cascade impaction test was performed according to the method described in USP 41, Chapter 1 to measure the aerosol performance of the spray-dried formulation when delivered from a dry powder inhaler device. The cascade impactor used was a Next Generation Impactor (NGI; USP 41, Chapter 1). <601> ). For the aerosol measurements performed in these examples, one size 3 HPMC capsule containing the spray-dried powder formulation was dispersed from a spray-drying inhaler device and delivered to an NGI under a vacuum pulled at 60 L / min, according to USP methodology. Samples were collected from each stage of the NGI and assayed for protein content by UV absorbance at 280 nm. The key aerosol performance parameters calculated from these measurements were a) fine particle fraction <5 μm (FPF<5 μm) (defined as the fraction of powder emitted from the device that has a measured aerodynamic particle size of <5 μm), and b) mass median aerodynamic diameter MMAD.

[0216] [Table 7]

[0217] A summary of the aerosol analysis results is summarized in Table 8.

[0218] [Table 8]

[0219] Example 3 - Saturation Velocity Modeling To understand the crystallinity results in Example 1, modeling was performed to evaluate the saturation rates of leucine and trileucine during the expected particle formation process in a spray dryer. The modeling revealed that leucine crystallization on the surface is highly dependent on the concentration ratio of the two additives.

[0220] At a leucine:trileucine concentration ratio of 3.8, the saturation curves for both additives overlap, resulting in co-saturation during particle formation (Figures 9A-9B). This inhibits leucine crystallization, even when leucine is present at high concentrations. Particle formulations with high leucine concentrations (>15 mg / mL) and low trileucine concentrations (<1 mg / mL) showed increased particle crystallinity.

[0221] Example 4 - Evaluation of the effect of raw material factors on the physical properties of leucine / trileucine formulations A second set of leucine:trileucine formulations was generated to evaluate the effect of buffer wt% across a range of feedstock concentrations, to evaluate the effect of total leucine:trileucine wt%, and to evaluate the effect of various concentrations of leucine / trileucine at a set ratio of 3.8 (leucine:trileucine).

[0222] A total of 27 powders were produced, varying in tri-leucine, leucine, trehalose, and trehalose wt%. Three factors were evaluated at three levels to create a full factorial study design, as outlined in Table 9. The three factors were citrate buffer wt%; total ingredient concentration; and total tri-leucine and leucine concentration. All formulations were held constant at a leucine-to-tri-leucine ratio of 3.8.

[0223] [Table 9]

[0224] The 27 raw materials used are listed in Table 10. Each raw material was prepared by dissolving the excipients trehalose, trisodium citrate, leucine, and trileucine in water. Once all excipients were completely dissolved, the raw materials were spray-dried using the following process parameters: outlet temperature, 70°C; raw material feed rate, 18 ml / min; nebulizer gas flow rate, 13 kg / h; and drying gas flow rate, 155 kg / hr. These parameters were selected to achieve the target particle and aerosol characteristics of the dry powder formulations for inhalation. Each of the 27 formulations was manufactured in 65 g batch sizes to obtain sufficient powder for characterization. Batches were randomly assigned and manufactured over several days.

[0225] [Table 10]

[0226] All 27 formulations were tested for the following physical powder properties:

[0227] [Table 11]

[0228] The results showed that the total concentration and wt% of leucine and trileucine had the most significant effect on particle properties, with raw material concentration appearing to be a secondary factor. The results are summarized in Table 12.

[0229] [Table 12]

[0230] Example 5 - Production of an inhalable leucine / trileucine formulation containing an anti-interleukin-4 antibody binding fragment (Fab) The following example explores whether a biologically active ingredient could be included in the leucine / trileucine formulation system described above to create a product suitable for inhaled delivery. Described here is the use of an anti-interleukin-4 antibody binding fragment (Fab), which is a variable antigen-binding region of an IgG1 monoclonal that specifically binds to IL-4. The anti-IL-4 Fab was formulated as a spray-dried powder for inhaled delivery according to the mass concentrations outlined in Table 13.

[0231] [Table 13]

[0232] The anti-IL-4 Fab was first liquid formulated at a concentration of approximately 50 mg / mL in 105 mM trehalose, 30 mM citric acid, pH 6.0. Leucine, trileucine, trehalose, and citric acid were dissolved in separate aqueous solutions and then added to the anti-IL-4 Fab solution to create the bulk liquid feedstock solution for spray drying. Table 14 summarizes the feedstock compositions prepared to produce the target powder formulation compositions. The liquid feedstock solution was then spray-dried using the following process parameters: outlet temperature, 70°C; feedstock feed rate, 12 ml / min; nebulizer gas flow rate, 13 kg / hr; and drying gas flow rate, 130 kg / hr, which were selected to achieve the target particle and aerosol characteristics of the dry powder formulation for inhalation.

[0233] [Table 14]

[0234] A summary of the powder and aerosol performance characterization results for the spray-dried formulations are summarized in Table 15. Powder and aerosol characterization was performed according to the specified methodology. The aerosol performance of the 14.5 w / w% and 40 w / w% anti-IL-4 Fab spray-dried formulations was tested at 65 mg and 50 mg of powder, which were filled into size 3 HPMC capsules and dispensed from a dry powder inhaler device.

[0235] [Table 15]

[0236] The results show that a spray-dried formulation containing a 5.25:1 ratio of leucine and trileucine was effective in achieving the target powder properties for two different doses of anti-IL-4 Fab. 3 ) was observed, and a low specific surface area (4.52 m 2 / g) was also observed, and at the same time, very high powder dispersibility (MMAD 3.0-3.1 μm; FPF < 5 μm of approximately 60-70%) was also achieved.

[0237] Example 6 - Production of an inhalable leucine / trileucine formulation containing an anti-TSLP antibody binding fragment (Fab) The properties of other formulations containing different Fabs were tested. Anti-TSLP Fabs derived from human IgG1 monoclonal antibodies that specifically bind to TSLP (thymic stromal lymphopoietin) were used (see sequences set forth in SEQ ID NOS: 1-8 provided herein). Separate formulations were generated containing the mass concentrations outlined in Table 16.

[0238] [Table 16]

[0239] The anti-TSLP Fab was initially received in a liquid buffer containing 105 mM trehalose, 30 mM citric acid, pH 6.0. Leucine, trileucine, trehalose, and citric acid were dissolved in separate aqueous solutions and then added to the anti-TSLP Fab solution to create the bulk liquid feedstock solution for spray drying. Table 17 summarizes the feedstock compositions prepared to achieve the target powder formulation compositions. The liquid feedstock solutions were then spray-dried using the process parameters listed in Table 18. The parameters were selected to achieve the target particle and aerosol properties of the dry powder formulation for inhalation.

[0240] [Table 17]

[0241] [Table 18]

[0242] A summary of the powder and aerosol performance characterization results for the spray-dried formulations are summarized in Table 19. For aerosol performance measurements, all three formulations were tested with 20 mg of spray-dried powder filled into size 3 HPMC capsules and dispensed from a dry powder inhaler device.

[0243] [Table 19]

[0244] Of particular note is the successful filling of 50 mg of Formulation #3 into a single Size 3 HPMC capsule due to the high bulk density of the powder. The high compressed bulk density (CBD) allowed for the delivery of a very high payload from a single capsule (approximately 14 mg FPM<5 μm, 82% FPF, 2.4 μm MMAD).

[0245] Additionally, Formulation #3 exhibited similar CBD (0.58 g / cm) and SSA (4.6 m / g) to anti-IL-4 Fab Formulation #2 (cBD = 0.59 g / cm, SSA = 4.5 m / g), suggesting that this powder profile is transferable between pharmaceutical formulations containing different active ingredients of the same class of molecules.

[0246] Example 7 - Powder and aerosol properties of spray-dried anti-TSLP formulations at three batch sizes This example provides an analysis of the powder and aerosol properties of an anti-TSLP Fab leucine / trileucine formulation using larger batch sizes to enable non-GLP and GLP inhalation toxicity testing. Scale-up requires the use of alternative scale spray dryer equipment and adjustment of spray drying process parameters to account for the increased heat and mass passing through the system and the need for longer processing runs.

[0247] Three batches of spray-dried anti-TSLP Fab formulation were produced with increasing batch sizes. The batches contained the following: anti-TSLP Fab 40 w / w%, trehalose 39 w / w%, leucine 10.5 w / w%, trileucine 2 w / w%, and citric acid pH 6.0 8.5 w / w%. The process parameters selected for each batch are shown in Table 20.

[0248] [Table 20]

[0249] Aerosol performance testing of batch #1 was performed at a powder fill mass of 50 mg in size 3 HPMC capsules, while batches #2 and #3 were tested at a fill mass of 20 mg. As batch size increased from 8.5 g to 1.2 kg, there was a slight increase in oVMD. For batch 3, it was 0.45 to 0.85 g / cm. 3A compacted bulk powder density (cBD) of 0.001 was achieved. Due to the high payload delivery of anti-TSLP Fab from the capsule-based inhaler device, the aerosol performance of the powder was also maintained independent of batch size, demonstrating the scalability of the formulation with minimal adjustments to the spray dryer process. A summary of all the results of the powder characterization and aerosol performance testing is summarized in Table 21.

[0250] [Table 21]

[0251] Example 8 Further Characterization of Leucine / Trileucine Formulations with Surfactants Additional batches of trileucine / leucine formulations containing various amounts of PS-80 were produced. The formulation compositions and process parameters for the production of each batch are shown in Table 22. The formulations were otherwise produced as described in Example 6.

[0252] [Table 22]

[0253] The aerosol properties of the formulations in Table 22 were analyzed using the method disclosed in Example 7. The results of this analysis are shown in Table 23.

[0254] [Table 23]

[0255] The aggregate content, oVMD, residual moisture content, Tg, cBD, and SSA were also measured using the methods described in the previous examples. The results of the powder characterization are shown in Table 24.

[0256] [Table 24]

[0257] This analysis shows that regardless of the % (w / w) amount of PS-80, the powder properties are similar to the control formulation.

[0258] The formulations described in Table 22 were then analyzed for subvisible particle (SVP) content. Subvisible particle (SVP) content was measured using micro-flow imaging technology (MFI). MFI combines microfluidic flow microscopy with high-resolution imaging particle analysis to quantify SVP counts and bin these counts across a range of particle sizes. Prior to testing, powder samples were dissolved in water, gently swirled to ensure uniform particle dispersion, and then loaded into a Protein Simple MFI 5200 (CA, USA). Results were reported as the number of various particle sizes (≦1 μm, ≦2 μm, ≦5 μm, ≦10 μm, and ≦25 μm) per ml. Figure 10A shows that the inclusion of 0.27% (w / w) PS-80 in the dry powder formulation reduces the absolute number of SVPs per ml upon reconstitution. This reduction in SVP count is attenuated by increasing the concentration of PS-80. Upon addition of 0.67% (w / w) PS-80, a significant decrease in SVP was observed, with negligible amounts of SVP with particle sizes greater than 5 μm. This trend was observed when the formulation was reconstituted to a concentration of 30 mg / ml FAB1 or 2.5 mg / ml FAB1 (Figure 10B).

[0259] Formulation characterization and SVP analysis were performed as described above for the second excipient-containing formulation, which used poloxamer 188 as the excipient rather than PS-80.

[0260] Multiple w / w% amounts of poloxamer 188 were investigated. The formulation compositions and process parameters for the production of each formulation batch were as described in Table 22 for the PS-80-containing formulations. The amount of trehalose was varied to compensate for the variable amount of poloxamer 188.

[0261] Aggregate content, oVMD, residual moisture content, Tg, cBD, and SSA were also measured using the methods described in the previous examples. The results of the powder characterization are shown in Table 25.

[0262] [Table 25]

[0263] The aerosol properties of the poloxamer-188 formulations were also analyzed using the method disclosed in Example 7. The results are shown in Table 26.

[0264] [Table 26]

[0265] P188 formulations were analyzed for SVP content using the method described above. Figure 11A shows that inclusion of 0.67% (w / w) P188 in the dry powder formulation reduces the absolute number of SVPs per ml upon reconstitution. This trend was observed when the formulations were reconstituted to a concentration of 30 mg / ml FAB1 (Figure 11A) or 2.5 mg / ml FAB1 (Figure 11B).

[0266] Example 9 Characterization of Leucine / Tri-Leucine Formulations Containing 1.1% (w / w) PS-80 In this example, the powder properties of dry powder formulations containing 1% or 40% (w / w) Fab1 and 1.1% (w / w / ) PS-80 were analyzed. The complete composition of the formulations is shown in Table 27. The formulations were prepared as described in Example 6.

[0267] [Table 27]

[0268] The stability of the formulations was analyzed after storage for 1 or 3 months at 40°C and 75% relative humidity (40 / 75) or 25°C and 60% relative humidity (25 / 60). Particle size distribution, moisture content, and surface roughness were examined. Figures 12A and 12B show that the moisture content and particle size distribution remained stable over time for a formulation containing 40% (w / w) Fab1. Figure 12C shows that particle morphology remained constant over time. Figures 13A and 13B show that the moisture content and particle size distribution remained stable over time for a formulation containing 1% (w / w) Fab1. Figure 13C shows that particle morphology remained constant over time.

[0269] SVP formation was analyzed upon reconstitution after storage at 40 / 75 for 1 month or 3 months or at 25 / 60 for 3 months. Analysis was performed as described in Example 9. Figure 14A shows that the amount of SVP produced under each condition upon reconstitution of a 40% (w / w) Fab1 formulation to a Fab1 concentration of 30 mg / ml remains unchanged. Figure 14B shows that the amount of SVP produced under each condition upon reconstitution of a 1% (w / w) Fab1 formulation to a Fab1 concentration of 0.75 mg / ml remains unchanged.

[0270] Aerosol properties were also tested after storage, and the results are shown in Tables 28 and 29.

[0271] [Table 28]

[0272] [Table 29]

[0273] The percent delivered dose (DD) was also determined after storage of each formulation under each condition, and the results are shown in Tables 28 and 29.

[0274] The potency of Fab1 in each of the formulations described in Table 27 was also tested after storage at 40 / 75 for 1 month or 3 months or at 25 / 60 for 3 months.

[0275] The potency was determined using homogeneous time-resolved fluorometry (HTRF). HTRF combines fluorescence resonance energy transfer (FRET) and time-resolved fluorescence (TR) techniques. When two fluorophores (donor and acceptor) are close to each other, excitation of the donor promotes energy transfer to the acceptor, resulting in a FRET signal. In this assay, streptavidin-europium cryptate bound to biotinylated human TSLP is the donor, and d2-labeled anti-TSLP mAb is the acceptor. FAB1 binds to human TSLP and prevents binding of the labeled mAb. This, in turn, increases the distance between the donor and acceptor fluorophores, resulting in a decrease in the FRET signal.

[0276] After assessing parallelism between the reference standard and the assay control or between the reference standard and the test samples, a constrained four-parameter logistic (4PL) curve fit is performed and the relative potencies of the FAB1 assay control and test samples are calculated by dividing the IC50 value of the reference standard by the IC50 value of the assay control or each test sample and multiplying by 100%.

[0277] The potency levels of Fab1 were 85-110% of the potency of Fab1 immediately after reconstitution from comparable formulations (i.e., t=0).

[0278] While certain embodiments have been illustrated and described herein, it is to be understood that the claims are not limited to the specific forms or arrangements of elements that have been described and illustrated. Although exemplary embodiments have been disclosed and specific terms are employed herein, they are used in a generic and descriptive sense only and not for purposes of limitation. Modifications and variations of the embodiments are possible in light of the above teachings. It is therefore to be understood that the embodiments may be practiced otherwise than as specifically described.

[0279] While various embodiments have been described above, it should be understood that they are presented merely as illustrations and examples of the present technology, and not as limitations. It will be apparent to those skilled in the relevant art that various changes in form and detail can be made therein without departing from the spirit and scope of the present technology. Therefore, the breadth and scope of the present technology should not be limited by any of the above-described embodiments, but should be defined only in accordance with the appended claims and their equivalents. It will also be understood that each feature of each embodiment described herein and each reference cited herein can be used in combination with the features of any other embodiment. All patents and publications discussed herein are incorporated herein by reference in their entirety.

Claims

1. 1. A dry powder formulation comprising a plurality of microparticles, the microparticles comprising leucine, trileucine and an active agent, wherein the leucine and trileucine are present in a ratio of leucine:trileucine of 0.1:1 to 30:1 by weight.

2. 2. The dry powder formulation of claim 1, wherein the ratio of leucine to trileucine is 0.1:1 to 25:1, 0.5:1 to 20:1, 1:1 to 20:1, 1:1 to 15:1, 1:1 to 12:1, 1:1 to 10:1, 1:1 to 7:1, 1:1 to 6:1, or 1:1, 2:1, 3:1, 4:1, 5:1, 5.1:1, 5.2:1, 5.25:1, 5.3:1, 5.4:1, 5.5:1, 5.75:1, or 6:1 leucine:trileucine.

3. 3. The dry powder formulation of claim 1 or 2, wherein the ratio of leucine:trileucine is 5.25:

1.

4. The dry powder formulation has a compressed bulk density of 0.4 to 1.0 g / cm ３ The dry powder formulation according to any one of claims 1 to 3, wherein

5. 5. The dry powder formulation of any one of claims 1 to 4, further comprising a glass stabilizer.

6. 6. The dry powder formulation of claim 5, wherein the glass stabilizer is an amorphous sugar or a buffering agent.

7. 6. The dry powder formulation of claim 5, wherein the glass stabilizer comprises an amorphous sugar and a buffering agent.

8. 8. The dry powder formulation of claim 6 or 7, wherein the amorphous sugar is selected from the group consisting of trehalose, sucrose, raffinose, inulin, dextran, mannitol, and cyclodextrin.

9. 9. The dry powder formulation of claim 6, wherein the buffer is selected from the group consisting of a citrate buffer, a phosphate buffer, a histidine buffer, a glycine buffer, an acetate buffer, and a tartrate buffer.

10. The dry powder formulation according to any one of claims 6 to 9, wherein the amorphous sugar is trehalose.

11. The dry powder formulation of any one of claims 1 to 10, wherein the active agent is an antibody or an antigen-binding fragment thereof.

12. 11. The dry powder formulation of any one of claims 1 to 10, further comprising a surfactant, optionally selected from polysorbate-20 (PS-20), polysorbate-40 (PS-40), polysorbate-60 (PS-60), polysorbate-80 (PS-80), and poloxamer-188.

13. 13. The dry powder formulation of claim 12, wherein the surfactant is PS-80, and optionally the PS-80 is present at a concentration ranging from 0.27% to 2.7% by weight, optionally at a concentration ranging from 0.67% to 1.33% by weight.

14. 13. The dry powder formulation of claim 12, wherein PS-80 is present at a concentration of 1.1% by weight.

15. Compressed bulk density is 0.5 g / cm ３ ~0.8g / cm ３ The dry powder formulation of any one of claims 1 to 14, wherein

16. 16. The dry powder formulation of any one of claims 1 to 15, having a maximum solids loading of 190 to 250 mg / mL.

17. The dry powder formulation of any one of claims 1 to 16, for administration by inhalation.

18. 18. The dry powder formulation of claim 17, in an aerosol form for delivery of the dry powder formulation to the lungs.

19. 20. The dry powder formulation of claim 18 for treating a medical condition in a mammalian patient.

20. 20. The dry powder formulation of claim 19, which is administered by a dry powder inhaler (DPI).

21. A method for preparing the dry powder formulation of any one of claims 1 to 20, comprising the steps of: a. preparing a liquid feedstock comprising leucine, trileucine, and an active agent, wherein the leucine and trileucine are present in a ratio of leucine:trileucine of 0.1:1 to 30:1 by weight; b. Atomizing the liquid feedstock; and c. Spray-drying the atomized liquid feedstock to form a plurality of microparticles. A method comprising:

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