3D printing of biologics

Abstract

The present disclosure provides pharmaceutical compositions and methods of preparing pharmaceutical compositions containing an amino acid-based biologic through the use of 3D printing. These pharmaceutical compositions may be used in the treatment of a disease or disorder.

Description

DESCRIPTION 3D PRINTING OF BIOLOGICS

[0001] This application claims the benefit of priority to United States Provisional Application No. 63 / 579,852, filed August 31, 2023, the contents of which are hereby incorporated by referenced. BACKGROUND 1. Field

[0002] The present disclosure relates generally to the field of laser based additive manufacturing of biologics. More particularly, it concerns compositions and methods relating to a powder bed additive manufacturing process of biological molecules. 2. Description of Related Art

[0003] 3D printing of personalized medication and pharmaceutical dosage form is a current and promising field of research. In the last decade, this topic of research has attracted the attention of multiple research groups and its potential has been exploited by a few pharmaceutical industries. 3D printing or additive manufacturing can be defined as a process of making 3-dimensional objects from a digital file. Even though the process of 3D printing can be defined by its final product, multiple technologies work on different principles to manufacture said 3-dimensional objects. Manufacturing of 3D printed objects using a powder bed is one of these approaches. Powder bed-based 3D printing platforms have two chambers, namely the reservoir chamber and a print chamber. The reservoir chamber is filled with the powder which is used to prepare the object. This bulk of powder is transferred to the built chamber layer-by-layer where it is exposed to a stimulus in a pattern fed by the software and the digital file, forming a 3-dimensional object. Platforms operating on this mechanism include Selective laser Sintering (SLS) based 3D printing, and Binder Spraying / Jetting based 3D printing.

[0004] Both selective laser sintering and binder jetting 3D printing require powders which have excellent flow characteristics for the process to function smoothly and print to be successful. In other fields of manufacturing where the powder usually comprises a singlecomponent, these processes are easy to optimize and implement. In contrast, pharmaceutical dosage forms usually contain more than one component which complicates the process for pharmaceutical 3D printing. Apart from the active pharmaceutical ingredients, components in a pharmaceutical dosage form include processing or performance aids. Processing aids such as binders, lubricants, glidants, fillers, and taste-masking agents, whereas the performance aids include release controlling agents, release modifying agents, and pH modifying agents. Processing a physical blend with multiple components can be challenging and can lead to several manufacturing issues such as print failure, nonuniformity of drug content, product variability (weight, %assay, dimensions), performance variability (dissolution rate, disintegration time).

[0005] Conventional formulations of biologics often require the need for solvents which has the potential to cause instability in biologic formulations. The printing of biologics circumvents that need for the use of solvents and hence has the capability of enhancing the stability of the formulation. The selective laser sintering of biologics is yet to be reported in literature. 2 4865-0355-2478, v. 1SUMMARY

[0006] The present disclosure encompasses a novel additive manufacturing process wherein biologics are used as the active pharmaceutical ingredient in a selective laser sintering process. In particular, the present disclosure provides methods of three dimensional printing of biologics which show enhanced stability with the ability to form complex three dimensional structures using powder-based 3D printing technologies for the manufacturing of pharmaceutical dosage forms. The invention encompasses a novel additive manufacturing process wherein biologics are used as the active pharmaceutical ingredient in a selective laser sintering process. In some embodiments, the biologic molecule is physically mixed with a polymer capable of melting and fed manually or mechanically into the powder bed of the selective laser sintering printer.

[0007] In some aspects, the present disclosure provides methods of preparing a pharmaceutical composition comprising: (A) obtaining a composition comprising: (i) an amino acid based biologic active pharmaceutical ingredient; and (ii) a polymer, wherein the polymer is in the semi-crystalline or crystalline state; (B) applying the composition to a surface of a chamber; wherein the chamber has been set to a first temperature; and (C) melting the composition with a laser to obtain the pharmaceutical composition.

[0008] In some embodiments, the amino acid based biologic active pharmaceutical ingredient is a peptide. In other embodiments, the amino acid based biologic active pharmaceutical ingredient is a protein. In other embodiments, the amino acid based biologic active pharmaceutical ingredient is an antibody.

[0009] In some embodiments, the composition comprises an amount from about 1% w / w to about 90% w / w of the amino acid based biologic active pharmaceutical ingredient. In some embodiments, the amount of the amino acid based biologic active pharmaceutical ingredient is from about 2.5% w / w to about 50% w / w of the composition. In some embodiments, the amount of the amino acid based biologic active pharmaceutical ingredient is from about 5% w / w to about 25% w / w of the composition. In some embodiments, the amount of the amino acid based biologic active pharmaceutical ingredient is from about 5% w / w to about 25% w / w of the composition. 3 4865-0355-2478, v. 1

[0010] In some embodiments, the composition comprises an amount from about 10% w / w to about 99% w / w of the polymer. In some embodiments, the amount of the polymer is from about 50% w / w to about 97.5% w / w of the composition. In some embodiments, the amount of the polymer is from about 75% w / w to about 95% w / w of the composition. In some embodiments, the amount of the polymer is from about 85% w / w to about 95% w / w of the composition.

[0011] In some embodiments, the polymer is in a semi-crystalline state. In some embodiments, the polymer is in a crystalline state. In some embodiments, the polymer comprises one or more blocks of polyethylene oxide or polypropylene oxide. In some embodiments, the polymer comprises one or more polyethylene oxide blocks. In some embodiments, the polymer is a diblock or a triblock copolymer. In some embodiments, the polymer is a triblock copolymer. In some embodiments, the triblock copolymer comprises two polyethylene glycol blocks. In some embodiments, the triblock copolymer comprises a polypropylene glycol block. In some embodiments, the triblock copolymer comprises two polyethylene glycol blocks on either side of a polypropylene glycol block. In some embodiments, the triblock copolymer comprises one or more polyethylene glycol block with a number of repeating units from 60 to about 140. In some embodiments, the number of repeating units in the polyethylene glycol block is from about 80 to about 120. In some embodiments, the triblock copolymer comprises a polypropylene glycol block with a number of repeating units from about 30 to about 80 repeating units. In some embodiments, the number of repeating units in the polypropylene glycol block is from about 50 to about 60. In some embodiments, the polymer is a triblock copolymer comprising two polyethylene glycol with a number of repeating units from about 80 to about 120 and a polypropylene glycol with a number of repeating units from about 50 to about 60.

[0012] In some embodiments, the laser is a CO2 laser. In some embodiments, the methods comprise using a laser with a speed from about 25 mm / s to about 300 mm / s. In some embodiments, the speed is from about 50 mm / s to about 250 mm / s. In some embodiments, the speed is from about 100 mm / s to about 200 mm / s. In some embodiments, the methods comprise a hatch spacing between the laser from about 10 µm to about 1 mm. In some embodiments, the hatch spacing is from about 50 µm to 500 µm. In some embodiments, the hatch spacing is from about 100 µm to 300 µm.

[0013] In some embodiments, the first temperature is below 100 °C. In some embodiments, the first temperature is from about −120 °C to about 80 °C. In some embodiments, the first temperature is from about −100 °C to about 60 °C. In some 4 4865-0355-2478, v. 1embodiments, the first temperature is from about 0 °C to about 50 °C. In some embodiments, the methods comprise adjusting the temperature of the surface to a surface temperature that is distinct from the first temperature. In some embodiments, the surface temperature is from about −120 °C to about 80 °C. In some embodiments, the surface temperature is from about −100 °C to about 60 °C. In some embodiments, the surface temperature is from about 0 °C to about 50 °C. In some embodiments, the difference between the first temperature and the surface temperature is from about 0.1 °C to about 10 °C. In some embodiments, the difference is from about 0.5 °C to about 5 °C. In some embodiments, the laser is used in an additive manufacturing technique. In some embodiments, the additive manufacturing technique is a melting process. In some embodiments, the additive manufacturing technique is powder bed fusion. In some embodiments, the methods further comprise one or more processing steps. In some embodiments, the processing step is blending. In some embodiments, the processing step is milling. In some embodiments, the processing step is tablet pressing. In some embodiments, the processing step is coating.

[0014] In still yet another aspect, the present disclosure provides pharmaceutical compositions comprising (A) an amino acid based biologic active pharmaceutical ingredient; and (B) a pharmaceutically acceptable polymer, wherein the pharmaceutically acceptable polymer comprises at least 80% of the composition and the pharmaceutically acceptable polymer is in the semi-crystalline or crystalline state.

[0015] In some embodiments, the amino acid based biologic active pharmaceutical ingredient is a peptide. In other embodiments, the amino acid based biologic active pharmaceutical ingredient is protein. In other embodiments, the amino acid based biologic active pharmaceutical ingredient is an antibody.

[0016] In some embodiments, the composition comprises an amount from about 1% w / w to about 90% w / w of the amino acid based biologic active pharmaceutical ingredient. In some embodiments, the amount of the amino acid based biologic active pharmaceutical ingredient is from about 2.5% w / w to about 50% w / w of the composition. In some embodiments, the amount of the amino acid based biologic active pharmaceutical ingredient is from about 5% w / w to about 25% w / w of the composition. In some embodiments, the amount of the amino acid based biologic active pharmaceutical ingredient is from about 5% w / w to about 25% w / w of the composition. 5 4865-0355-2478, v. 1

[0017] In some embodiments, the composition comprises an amount from about 10% w / w to about 99% w / w of the polymer. In some embodiments, the amount of the polymer is from about 50% w / w to about 97.5% w / w of the composition. In some embodiments, the amount of the polymer is from about 75% w / w to about 95% w / w of the composition. In some embodiments, the amount of the polymer is from about 85% w / w to about 95% w / w of the composition.

[0018] In some embodiments, the polymer is in a semi-crystalline state. In some embodiments, the polymer is in a crystalline state. In some embodiments, the polymer comprises one or more blocks of polyethylene oxide or polypropylene oxide. In some embodiments, the polymer comprises one or more polyethylene oxide blocks. In some embodiments, the polymer is a diblock or a triblock copolymer. In some embodiments, the polymer is a triblock copolymer. In some embodiments, the triblock copolymer comprises two polyethylene glycol blocks. In some embodiments, the triblock copolymer comprises a polypropylene glycol block. In some embodiments, the triblock copolymer comprises two polyethylene glycol blocks on either side of a polypropylene glycol block. In some embodiments, the triblock copolymer comprises one or more polyethylene glycol block with a number of repeating units from 60 to about 140. In some embodiments, the number of repeating units in the polyethylene glycol block is from about 80 to about 120. In some embodiments, the triblock copolymer comprises a polypropylene glycol block with a number of repeating units from about 30 to about 80 repeating units. In some embodiments, the number of repeating units in the polypropylene glycol block is from about 50 to about 60. In some embodiments, the polymer is a triblock copolymer comprising two polyethylene glycol with a number of repeating units from about 80 to about 120 and a polypropylene glycol with a number of repeating units from about 50 to about 60.

[0019] In still yet another aspect, the present disclosure provides methods of treating or preventing a disease or disorder comprising administering a pharmaceutical composition described herein to a patient in need thereof, wherein the pharmaceutical composition comprises an amino acid based active pharmaceutical ingredient capable of treating or preventing the disease or disorder.

[0020] The use of the word “a” or “an” when used in conjunction with the term “comprising” in the claims and / or the specification may mean “one,” but it is also consistent with the meaning of “one or more,” “at least one,” and “one or more than one.” The word “about” means plus or minus 5% of the stated number. 6 4865-0355-2478, v. 1

[0021] It is contemplated that any method or composition described herein can be implemented with respect to any other method or composition described herein. Other objects, features and advantages of the present disclosure will become apparent from the following detailed description. It should be understood, however, that the detailed description and the specific examples, while indicating specific embodiments of the disclosure, are given by way of illustration only, since various changes and modifications within the spirit and scope of the disclosure will become apparent to those skilled in the art from this detailed description. 7 4865-0355-2478, v. 1BRIEF DESCRIPTION OF THE DRAWINGS

[0022] The following drawings form part of the present specification and are included to further demonstrate certain aspects of the present disclosure. The disclosure may be better understood by reference to one or more of these drawings in combination with the detailed description of specific embodiments presented herein.

[0023] FIG.1 shows the process schematic for the manufacturing of the biologics.

[0024] FIG.2 shows circular dichroism of the printed construct to analyze the stability of the biologic molecule within the structure.

[0025] FIG. 3 shows an analysis of the circular dichroism spectra to understand the secondary structure of the biologic.

[0026] FIG. 4 shows a Fourier transform characterization of the biologic to confirm stability of biologics post laser exposure. DESCRIPTION OF ILLUSTRATIVE EMBODIMENTS

[0027] The present disclosure relates to the laser based additive manufacturing of biologics. Briefly, the present disclosure comprises a powder bed based additive manufacturing process based on the selective fusion of the material to form 3D structures. In some embodiments, this process is a powder bed fusion process which involves a powder bed-based feedstock layer being selectively fused, utilizing electromagnetic radiation and thermal stimulus. In some embodiments, the present disclosure may involve the principle of heating up a solid and fusing it on the surface to form consolidated structures (Davis et al., 2021; Thakkar et al., 2021; Fina et al., 2017). I. Biologics A. Production

[0028] Biologic molecules or biologics are produced from a living organism or component of a living organism and are capable of being used as therapeutics. In the U.S. alone, biologics account for 40% of prescription drug spending, despite only 2% of the population using biologic drugs (Mulcahy et al., 2018). The cost of these medications is quicklyrising from expensive to simply unaffordable. This global issue will likely become more problematic with the steady growth of the world population and the advent of new biologic therapies. Some strategic measures have been implemented to mediate the rising prices such as a cost-effectiveness analysis methodology which appraises health interventions and selects technologies based on the returns expected for a financial investment, therefore urging pharmaceutical companies to provide biologics at competitive rates. Many governments have passed legislature aimed at reducing expenditures on follow-on biologics such as biosimilars which decreases the costs of a product entering the market by shortening the approval pathway, ensuring compulsory licensing and encouraging data sharing. These strategic initiatives are expected to moderate the high costs of new biologics.

[0029] In some embodiments, the biologic described herein may be derived from a standard manufacturing scheme for biologics. In some embodiments, this scheme may be optimized for fed-batch bioreactors that are well adapted for suspension cell culture processes, followed by a combination of different filtration and chromatography unit operations to achieve the target purity requirement and yield. Both fed-batch bioreactors and resin column chromatography operations experience an over-sizing problem when scaled up to pilot and manufacturing process scales. Fed-batch reactors, which comprise about 90% of commercial biologics production, require large-volume tanks to counter the low productivity per unit volume (Rose et al., 2003) and resin chromatography columns, limited by intrinsically slow mass transfer rates, have to be over-sized in order to operate at a higher flow rate to achieve desirable productivity (Reichert et al., 2002). This approach of attaining better productivity via enlarged working units occurs at the expense of process economics and flexibility.

[0030] Although monoclonal antibody (mAb) production has experienced improvements from some single-use (SU) upstream and downstream technologies, strategies are required to combine and implement these advances for more efficient and economical production. Typical mAb manufacturing practices involve several stages of inoculum development in small reactors followed by cell cultivation in larger stainless steel bioreactors (5000 L to 25,000 L). The WAVE bioreactors, now offered by GE Healthcare, were the first SU bioreactors designed for large scale manufacturing. Introduced in 1996, the WAVE bioreactor consisted of a plastic bag on rocker platform that provides agitation and gas transfer. Despite the evolution over the years, the WAVE bioreactor is only available in volume sizes 9 4865-0355-2478, v. 1of up to 500 L and therefore usually operated as part of seed expansion in mammalian cell culture-based bio-therapeutics manufacturing (Eibl et al., 2010).

[0031] For an upstream process using fed-batch cell culture, process scale-up can be achieved by a faster cell culture production rate using multiple bioreactors that are harvested at a certain frequency per week. A more common scale-up method to increase manufacturing capability is via larger working vessels, which requires developing a dedicated bioprocess facility through expansion or new construction (Shimoni et al., 2013). For example, for commercial scale mAb production, the traditional fed-batch bioreactor usually cultures cells in 10,000–25,000 L stainless steel tanks for 7–21 days with a product yield of 2–6 g / L (Rose et al., 2003).

[0032] In some aspects, different chromatography techniques (affinity, ion exchange, hydrophobic interaction) may be implemented in biologics purification in the format of resin columns. The majority of functional ligands are grafted within internal pores of polymeric chromatography resins. Simulated moving bed chromatography provides a way of continuous processing in a capture unit operation. The continuous purification of a capture target is aided by adoption of integrated valve modules, such as the valve manifold featured in the BioSMB™ chromatography system (Tarpon Biosystems, Inc.). The computer-assisted valve manifold can be programmed for automatic switching of column feeds between feed stream, wash buffer, elution buffer, cleaning buffer, and equilibration buffer, to create the simulated moving bed separation.

[0033] In some embodiments, the biologic described herein may be derived from an engineered cell. An engineered cell be derived from any mammalian organ or tissue, including the brain, nerves, ganglia, spine, eye, heart, liver, kidney, lung, spleen, bone, thymus, lymphatic system, skin, muscle, pancreas, stomach, intestine, blood, ovary, uterus, or testes. A cell may be derived from a donor (e.g., an allogeneic cell), derived from a subject (e.g., an autologous cell), or from another species (e.g., a xenogeneic cell). In an embodiment, a cell can be grown in cell culture, or prepared from an established cell culture line, or derived from a donor (e.g., a living donor or a cadaver). In an embodiment, a cell is genetically engineered. In another embodiment, a cell is not genetically engineered. A cell may include a stem cell, such as a reprogrammed stem cell, or an induced pluripotent cell. Exemplary cells include mesenchymal stem cells (MSCs), fibroblasts (e.g., primary fibroblasts). HEK cells (e.g., HEK293T), Jurkat cells, HeLa cells, retinal pigment epithelial (RPE) cells, HUVEC cells, NIH3T3 cells, CHO- 10 4865-0355-2478, v. 1K1 cells, COS-1 cells, COS-7 cells, PC-3 cells, HCT 116 cells, A549MCF-7 cells, HuH-7 cells, U-2 OS cells, HepG2 cells, Neuro-2a cells, and SF9 cells.

[0034] A cell included in a device or capsule may produce or secrete a therapeutic agent. In an embodiment, a cell included in a device or capsule may produce or secrete a single type of therapeutic agent or a plurality of therapeutic agents. In an embodiment, a device or capsule may comprise a cell that is transduced or transfected with a nucleic acid (e.g., a vector) comprising an expression sequence of a therapeutic agent. A nucleic acid introduced into a cell (e.g., by transduction or transfection) may be incorporated into a nucleic acid delivery system, such as a plasmid, or may be delivered directly. In an embodiment, a nucleic acid introduced into a cell (e.g., as part of a plasmid) may include a region to enhance expression of the therapeutic agent and / or to direct targeting or secretion, for example, a promoter sequence, an activator sequence, or a cell-signaling peptide, or a cell export peptide. A. Formulation

[0035] Because biologics are often prone to degradation through heat and fusion processes, they are often overlooked. The mechanisms of degradation include aggregation, denaturation and surface adsorption of proteins which often occur in aqueous media. Further in liquid media, proteins undergo either chemical degradation (e.g., deamidation or oxidation) or physical degradation (aggregation and precipitation) (Narasimhan et al., 2016). This degradation in aqueous media may be due to heat, agitation, pH changes or freezing. Further, biologics are also instable in the presence of aqueous media at room temperature. Hence, proteins are formulated in solid form to preserve protein native structure and extend shelf life (Chen et al., 2021). Most protein formulations attempt to avoid protein unfolding and hence process such as lyophilization, spray drying, thin film freeing and solvent casting are often used to produce dry biologic formulations (Geraldes et al., 2020). The present disclosure describes a method to minimize the degradation of biologics within the printing process and further stabilize them through the addition of polymers capable of undergoing the laser based additive manufacturing process. Polymers (e.g., Pluronic, PEG) can undergo the melting process and in turn capable of forming consolidated structures using laser based additive manufacturing 3D printing. A powder mixture containing the biologic and polymer are used as the feedstock material. Further excipients can be used to maintain protein stability such as polyols, surfactants, buffering agents, inorganic salts, antioxidants, bulking agents, sugars and protectants. The technology ultimately utilizes this solvent free additive manufacturing 11 4865-0355-2478, v. 1platform to print biologic dosage forms for reconstitution and our ability to control specific process parameters allows us to use powder bed fusion printing to manufacture consolidated structures containing biologic molecules. These biologics may be used for disease areas such as oncology, cardiovascular health, metabolic disease, infectious disease, neurology, autoimmune disease, and others. These and other aspects of the disclosure are described in detail below.

[0036] The current technologies utilizing biologics are lyophilization, spray drying, solvent casting and thin film freezing. These technologies are associated with certain levels of degradation due to the stresses associated with each process (Chen et al., 2021). The technology described by the present disclosure utilizes a solvent free additive manufacturing technique to produce dry solid biologic dosage forms. Laser based additive manufacturing processes have found their way into drug delivery research and the printing of dosage forms, but these are related to tablets and drug delivery devices. Previous research regarding printing of drug molecules primarily deals with small molecular weight drugs (Davis et al., 2021; Thakkar et al., 2021; Fina et al., 2017).

[0037] The printing of biologics using laser assisted additive manufacturing is still yet to be explored and hence requires investigation. Stresses undergone during the drying process have the potential to alter the native conformation of the protein. These processes can cause interactions among proteins as well as interaction of the protein with the surrounding environment which leads to an energetically unfavorable state and ultimately to further structural disruption. They can also disrupt the secondary, tertiary as well as quaternary structure of the protein which could lead to further denaturation (Chen et al., 2021).

[0038] Biologics are prepared in their solid form to maintain their stability and extend their shelf life as they undergo degradation in aqueous media (Chen et al., 2021). Considering that the technique being utilized is a solvent free process, the chances of water based degradation is minimized. Further biologic molecules also need to be gently swirled during reconstitution and this process can further lead to damage of the biologic molecule (Karri et al., 2014). The use of excipients during the process allows for reduced swirling to be applied to solubilize the biologic molecule in aqueous media and further reduces the chances of damage. 12 4865-0355-2478, v. 1

[0039] The current technology described by the present disclosure is solvent free, which is advantageous in the manufacturing of biologic products. The current technology also allows for use at room temperature to manufacture consolidated structures with biologics. Further the gives the ability to rapid prototype different dosage forms containing biologic molecules. Lastly, it is a high throughput process which leads to expedited processing times.

[0040] The exposure to heat still leads to slight degradation of the biologic being printed but this can be overcome by improving the selection of polymers being used for the process. Increased selection of polymers will allow for printing at lower temperatures and further reduce or eliminate the heat associated degradation. Building a new prototype which can house multiple laser sources or allow switching of laser sources can help eliminate this problem. Often single laser process only allows the sintering of limited materials. Interchangeability of the laser source allows for increased selection of materials for printing.

[0041] This process may be used for continuous manufacturing of films and other dosage forms containing biologic formulations. Furthermore, this process has the potential to be used for the development of oral dosage forms for biologics. II. 3D Printing

[0042] The inventors contemplate using 3D printing processes that can fabricate 3D materials. Three-dimensional (3D) printing, also known variously as additive manufacturing (AM), layered manufacturing, rapid prototyping (RP), or solid freeform fabrication, represents the direct fabrication of parts layer-by-layer, guided by digital information from a computer- aided design (CAD) file without any part-specific tooling. In 3D printing, CAD models of parts to be manufactured are first sliced in a virtual environment to create a stack of two-dimensional (2D) slices. A 3D printing machine then builds the parts one layer at a time based on the 2D slice information, stacking and joining successive layers to make the final 3D object. With the advent of 3D printing, such virtual designs can now be rendered into physical 3D objects that can serve as prototypes or be directly used as functional parts for a variety of applications.

[0043] Three-dimensional printing technology offers significant advantages for biomedical devices and TE due to the ability to manufacture low-volume or one-of-a-kind parts on-demand based on patient-specific needs. For example, surgical implants are currently manufactured by making a near-net-shape part via forging, casting, or machining operations, followed by specialized surface finishing or treatments for the desired surface, mechanical 13 4865-0355-2478, v. 1properties, and aesthetic effects. 3D printing methods include material jetting, material extrusion, light polymerization (such as DLP, SLA and CLIP), powder bed (including SLS, discussed below), laminated object manufacturing, powder fed (DED, EHLA) and wire (EBF).

[0044] Selective laser sintering, or SLS, is an additive manufacturing (AM) technique that uses a laser as the power and heat source to melt or sinter powdered material (typically nylon or polyamide), aiming the laser automatically at points in space defined by a 3D model, binding the material together to create a solid structure. It is similar to selective laser melting; the two are instantiations of the same concept but differ in technical details.

[0045] An additive manufacturing layer technology, SLS involves the use of a high power laser (for example, a carbon dioxide laser) to fuse small particles of plastic, metal, ceramic, or glass powders into a mass that has a desired three-dimensional shape. The laser selectively fuses powdered material by scanning cross-sections generated from a 3D digital description of the part (for example from a CAD file or scan data) on the surface of a powder bed. After each cross-section is scanned, the powder bed is lowered by one layer thickness, a new layer of material is applied. Because finished part density depends on peak laser power, rather than laser duration, a SLS machine may uses a pulsed laser. It is contemplated that the laser can be either a The SLS machine preheats the bulk powder material in the powder bed somewhat below its melting point, to make it easier for the laser to raise the temperature of the selected regions the rest of the way to the melting point.

[0046] In contrast with SLA and FDM, which most often require special support structures to fabricate overhanging designs, SLS does not need a separate feeder for support material because the part being constructed is surrounded by unsintered powder at all times. This allows for the construction of previously impossible geometries. Also, since the machine's chamber is always filled with powder material the fabrication of multiple parts has a far lower impact on the overall difficulty and price of the design because through a technique known as 'Nesting', where multiple parts can be positioned to fit within the boundaries of the machine. One design aspect which should be observed however is that with SLS it is 'impossible' to fabricate a hollow but fully enclosed element. This is because the unsintered powder within the element could not be drained.

[0047] The quality of printed structures depends on the various factors include powder properties such as particle size and shape, density, roughness, and porosity. Furthermore, the particle distribution and their thermal properties affect a lot on the flowability of the powder. 14 4865-0355-2478, v. 1Commercially-available materials used in SLS come in powder form and include, but are not limited to, polymers such as polyamides (PA), polystyrenes (PS), thermoplastic elastomers (TPE), and polyaryletherketones (PAEK). Polyamides are the most commonly used SLS materials due to their ideal sintering behavior as a semi-crystalline thermoplastic, resulting in parts with desirable mechanical properties. Polycarbonate (PC) is a material of high interest for SLS due to its high toughness, thermal stability, and flame resistance; however, such amorphous polymers processed by SLS tend to result in parts with diminished mechanical properties, dimensional accuracy and thus are limited to applications where these are of low importance. Metal materials are not commonly used in SLS since the development of selective laser melting.

[0048] Powder particles are typically produced by cryogenic grinding in a ball mill at temperatures well below the glass transition temperature of the material, which can be reached by running the grinding process with added cryogenic materials such as dry ice (dry grinding), or mixtures of liquid nitrogen and organic solvents (wet grinding) (Schmidt et al., 2012). The process can result in spherical or irregular shaped particles as low as five microns in diameter. Powder particle size distributions are typically gaussian and range from 15 to 100 microns in diameter, although this can be customized to suit different layer thicknesses in the SLS process. Chemical binder coatings can be applied to the powder surfaces post-process; these coatings aid in the sintering process and are especially helpful to form composite material parts such as with alumina particles coated with thermoset epoxy resin (Yang et al., 2018).

[0049] Sintering in SLS primarily occurs in the liquid state when the powder particles forms a micro-melt layer at the surface, resulting in a reduction in viscosity and the formation of a concave radial bridge between particles, known as necking (Kruth et al., 2005) due to the material's response to lower its surface energy. In the case of coated powders, the purpose of the laser is to melt the surface coating which will act as a binder. Solid state sintering is also a contributing factor, albeit with a much reduced influence, and occurs at temperatures below the melting temperature of the material. The principal driving force behind the process is again the material's response to lower its free energy state resulting in diffusion of molecules across particles.

[0050] In some embodiments, the laser may be a CO2laser. The most basic form of a CO2 laser consists of a gas discharge (with a mix close to that specified above) with a total reflector at one end, and an output coupler (a partially reflecting mirror) at the output end. The CO2 laser can be constructed to have continuous wave (CW) powers between milliwatts (mW) 15 4865-0355-2478, v. 1and hundreds of kilowatts (kW). It is also very easy to actively Q-switch a CO2laser by means of a rotating mirror or an electro-optic switch, giving rise to Q-switched peak powers of up to gigawatts (GW).

[0051] Because the laser transitions are actually on vibration-rotation bands of a linear triatomic molecule, the rotational structure of the P and R bands can be selected by a tuning element in the laser cavity. Prisms are not practical as tuning elements because most media that transmit in the mid-infrared absorb or scatter some of the light, so the frequency tuning element is almost always a diffraction grating. By rotating the diffraction grating, a particular rotational line of the vibrational transition can be selected. The finest frequency selection may also be obtained through the use of an etalon. In practice, together with isotopic substitution, this means that a continuous comb of frequencies separated by around 1 cm−1(30 GHz) can be used that extend from 880 to 1090 cm−1. The laser's output wavelength is affected by the particular isotopes contained in the carbon dioxide molecule, with heavier isotopes causing longer wavelength emission. III. Pharmaceutical Compositions

[0052] In some aspects, the present disclosure provides methods of preparing pharmaceutical compositions containing a biologic active pharmaceutical ingredient. These compositions may be used to prepare a pharmaceutical composition from a starting material such as a filament or powder that exhibits one or more favorable properties such as exhibiting a free-flowing property as an angle of repose, sufficient strength, sufficient stress, bend angle, diameter, viscosity, or Carr’s Index. The polymer and the active pharmaceutical ingredient may comprise the active pharmaceutical ingredient and the polymer in a molar ratio. The amount of the composition which contains the polymer is greater than about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, 96%, 97%, 98%, or 99%. The pharmaceutical composition comprises from about 10%, 20%, 30%, 40%, 50%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 92.5%, 95%, 96%, 97%, 97.5%, 98%, 98.5%, 99%, 99.5%, to about 99.9%, or any range derivable therein of the polymer.

[0053] These compositions may exhibit one or more free-flowing properties such as having a flowability as measured by the angle of repose of less than 25. These compositions may exhibit a flowability as measured by the angle of repose of less than about 25, less than about 27.5, less than about 30, less than about 32.5, less than about 35, less than about 37.5, or less than about 40. The flowability may be from about 25 to about 40, or from about 25 to 16 4865-0355-2478, v. 1about 30. The flowability may be from about 2, 4, 5, 6, 8, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, or any range derivable therein. The flowability of the pharmaceutical composition is measured by, the simplest method for the determination of the angle of repose is the “poured” angle. A funnel with a wide outlet is affixed at a distance of 10 cm above the bench, where a piece of paper is placed directly beneath the funnel. The granules are added while the funnel is closed. The contents flow through and collect on the paper. The diameter of the cone (D) and two opposite sides (l1+ l2) are measured with rulers. The angle of repose (θ) is calculated from the equation arc cos[D / (l1 + l2)]. The relationship between flow properties and angle of repose has been established. When the angle of repose is less than 25 degrees, the flow is said to be excellent; on the other hand, if the angle of repose is more than 40 degrees, the flow is considered to be poor. These pharmaceutical compositions may be present as agglomerations and used in either a batch, semi-continuous, continuous manufacturing process. The active pharmaceutical ingredient may act as a binder between the absorbent particles within the pharmaceutical composition.

[0054] In other aspects, the present pharmaceutical compositions may exhibit a mean or average particle size distribution greater than 25 µm, greater than 50 µm, or greater than 60 µm. In some embodiments, the pharmaceutical compositions exhibit a mean or average particle size from about 25 µm to about 500 µm, 30 µm to about 400 µm, 35 µm to about 350 µm, 40 µm to about 300 µm, 50 µm to about 250 µm, 50 µm to about 200 µm, 50 µm to about 150 µm, 55 µm to about 125 µm, or from about 60 µm to about 100 µm. The mean or average particle size of the pharmaceutical composition comprises from about 25 µm, 30 µm, 35 µm, 40 µm, 45 µm, 50 µm, 55 µm, 60 µm, 65 µm, 70 µm, 75 µm, 80 µm, 85 µm, 90 µm, 95 µm, 100 µm, 105 µm, 110 µm, 115 µm, 120 µm, 125 µm, 150 µm, 175 µm, 200 µm, 250 µm, 300 µm, 350 µm, 400 µm, 450 µm, 500 µm, 600 µm, 700 µm, 800 µm, 900 µm, to about 1000 µm, or any range derivable therein. The mean or average particle size of the pharmaceutical composition may be determined by mesh analysis using a sonic sifter. The particle size distribution of the dried granules can also be determined by a dry laser diffraction technique or scanning electron microscopy. IV. Methods of Treatment

[0055] Described herein are methods of preparing a pharmaceutical composition (or medicament) for the treatment of a disease. 17 4865-0355-2478, v. 1

[0056] In some embodiments, the disease is a proliferative disease. In an embodiment, the proliferative disease is cancer. A cancer may be an epithelial, mesenchymal, or hematological malignancy. A cancer includes primary malignant cells or tumors (e.g., those whose cells have not migrated to sites in the subject's body other than the site of the original malignancy or tumor) and secondary malignant cells or tumors (e.g., those arising from metastasis, the migration of malignant cells or tumor cells to secondary sites that are different from the site of the original tumor). In an embodiment, the cancer is a solid tumor (e.g., carcinoid, carcinoma or sarcoma), a soft tissue tumor (e.g., a heme malignancy), or a metastatic lesion, e.g., a metastatic lesion of any of the cancers disclosed herein. In an embodiment, the cancer is a fibrotic or desmoplastic solid tumor. In some embodiments, the tumor is a mesothelioma tumor.

[0057] Exemplary cancers that can be treated by the methods provided for herein include, but are not limited to, carcinoma, lymphoma, blastoma, sarcoma, and leukemia or lymphoid malignancies. In an embodiment, the cancer affects a system of the body, e.g., the nervous system (e.g., peripheral nervous system (PNS) or central nervous system (CNS)), vascular system, skeletal system, respiratory system, endocrine system, lymph system, reproductive system, or gastrointestinal tract. In some embodiments, cancer affects a part of the body, e.g., blood, eye, brain, skin, lung, stomach, mouth, ear, leg, foot, hand, liver, heart, kidney, bone, pancreas, spleen, large intestine, small intestine, spinal cord, muscle, ovary, uterus, vagina, or penis. More particular examples of such cancers include squamous cell cancer (e.g., epithelial squamous cell cancer), lung cancer including small-cell lung cancer, non-small cell lung cancer, adenocarcinoma of the lung and squamous carcinoma of the lung, cancer of the peritoneum, hepatocellular cancer, gastric or stomach cancer including gastrointestinal cancer, pancreatic cancer, glioblastoma, cervical cancer, ovarian cancer, liver cancer, bladder cancer, hepatoma, breast cancer, colon cancer, rectal cancer, colorectal cancer, endometrial cancer or uterine carcinoma, salivary gland carcinoma, kidney or renal cancer, prostate cancer, vulval cancer, thyroid cancer, hepatic carcinoma, anal carcinoma, penile carcinoma, as well as head and neck cancer.

[0058] Other examples of cancers include, but are not limited to: Acute Childhood Lymphoblastic Leukemia, Acute Lymphoblastic Leukemia, Acute Lymphocytic Leukemia, Acute Myeloid Leukemia, Adrenocortical Carcinoma, Adult (Primary) Hepatocellular Cancer, Adult (Primary) Liver Cancer, Adult Acute Lymphocytic Leukemia, Adult Acute Myeloid Leukemia, Adult Hodgkin's Disease, Adult Hodgkin's Lymphoma, Adult Lymphocytic 18 4865-0355-2478, v. 1Leukemia, Adult Non-Hodgkin's Lymphoma, Adult Primary Liver Cancer, Adult Soft Tissue Sarcoma, AIDS-Related Lymphoma, AIDS-Related Malignancies, Anal Cancer, Astrocytoma, Bile Duct Cancer, Bladder Cancer, Bone Cancer, Brain Stem Glioma, Brain Tumors, Breast Cancer, Cancer of the Renal Pelvis and Ureter, Central Nervous System (Primary) Lymphoma, Central Nervous System Lymphoma, Cerebellar Astrocytoma, Cerebral Astrocytoma, Cervical Cancer, Childhood (Primary) Hepatocellular Cancer, Childhood (Primary) Liver Cancer, Childhood Acute Lymphoblastic Leukemia, Childhood Acute Myeloid Leukemia, Childhood Brain Stem Glioma, Childhood Cerebellar Astrocytoma, Childhood Cerebral Astrocytoma, Childhood Extracranial Germ Cell Tumors, Childhood Hodgkin's Disease, Childhood Hodgkin's Lymphoma, Childhood Hypothalamic and Visual Pathway Glioma, Childhood Lymphoblastic Leukemia, Childhood Medulloblastoma, Childhood Non-Hodgkin's Lymphoma, Childhood Pineal and Supratentorial Primitive Neuroectodermal Tumors, Childhood Primary Liver Cancer, Childhood Rhabdomyosarcoma, Childhood Soft Tissue Sarcoma, Childhood Visual Pathway and Hypothalamic Glioma, Chronic Lymphocytic Leukemia, Chronic Myelogenous Leukemia, Colon Cancer, Cutaneous T-Cell Lymphoma, Endocrine Pancreas Islet Cell Carcinoma, Endometrial Cancer, Ependymoma, Epithelial Cancer, Esophageal Cancer, Ewing's Sarcoma and Related Tumors, Exocrine Pancreatic Cancer, Extracranial Germ Cell Tumor, Extragonadal Germ Cell Tumor, Extrahepatic Bile Duct Cancer, Eye Cancer, Female Breast Cancer, Gaucher's Disease, Gallbladder Cancer, Gastric Cancer, Gastrointestinal Carcinoid Tumor, Gastrointestinal Tumors, Germ Cell Tumors, Gestational Trophoblastic Tumor, Hairy Cell Leukemia, Head and Neck Cancer, Hepatocellular Cancer, Hodgkin's Disease, Hodgkin's Lymphoma, Hypergammaglobulinemia, Hypopharyngeal Cancer, Intestinal Cancers, Intraocular Melanoma, Islet Cell Carcinoma, Islet Cell Pancreatic Cancer, Kaposi's Sarcoma, Kidney Cancer, Laryngeal Cancer, Lip and Oral Cavity Cancer, Liver Cancer, Lung Cancer, Lymphoproliferative Disorders, Macroglobulinemia, Male Breast Cancer, Malignant Mesothelioma, Malignant Thymoma, Medulloblastoma, Melanoma, Mesothelioma, Metastatic Occult Primary Squamous Neck Cancer, Metastatic Primary Squamous Neck Cancer, Metastatic Squamous Neck Cancer, Multiple Myeloma, Multiple Myeloma / Plasma Cell Neoplasm, Myelodysplastic Syndrome, Myelogenous Leukemia, Myeloid Leukemia, Myeloproliferative Disorders, Nasal Cavity and Paranasal Sinus Cancer, Nasopharyngeal Cancer, Neuroblastoma, Non-Hodgkin's Lymphoma During Pregnancy, Nonmelanoma Skin Cancer, Non-Small Cell Lung Cancer, Occult Primary Metastatic Squamous Neck Cancer, Oropharyngeal Cancer, Osteo- / Malignant Fibrous Sarcoma, Osteosarcoma / Malignant Fibrous Histiocytoma, Osteosarcoma / Malignant Fibrous 19 4865-0355-2478, v. 1Histiocytoma of Bone, Ovarian Epithelial Cancer, Ovarian Germ Cell Tumor, Ovarian Low Malignant Potential Tumor, Pancreatic Cancer, Paraproteinemias, Purpura, Parathyroid Cancer, Penile Cancer, Pheochromocytoma, Pituitary Tumor, Plasma Cell Neoplasm / Multiple Myeloma, Primary Central Nervous System Lymphoma, Primary Liver Cancer, Prostate Cancer, Rectal Cancer, Renal Cell Cancer, Renal Pelvis and Ureter Cancer, Retinoblastoma, Rhabdomyosarcoma, Salivary Gland Cancer, Sarcoidosis Sarcomas, Sezary Syndrome, Skin Cancer, Small Cell Lung Cancer, Small Intestine Cancer, Soft Tissue Sarcoma, Squamous Neck Cancer, Stomach Cancer, Supratentorial Primitive Neuroectodermal and Pineal Tumors, T-Cell Lymphoma, Testicular Cancer, Thymoma, Thyroid Cancer, Transitional Cell Cancer of the Renal Pelvis and Ureter, Transitional Renal Pelvis and Ureter Cancer, Trophoblastic Tumors, Ureter and Renal Pelvis Cell Cancer, Urethral Cancer, Uterine Cancer, Uterine Sarcoma, Vaginal Cancer, Visual Pathway and Hypothalamic Glioma, Vulvar Cancer, Waldenstrom's Macroglobulinemia, Wilms' Tumor, pleural cancer, pleural metastatic disease, pleurisy, lung infection, viral pneumonia, bacterial pneumonia, idiopathic pulmonary fibrosis, acute respiratory distress syndrome, pleural thickening, pleural pseudotumor, pleural plaque, extrapleural hematoma, Castleman disease, hemangioendothelioma, splenosis, paramalignang effusion, pleural effusion, pneumothorax, hemothorax, reactive pleuritis. In some embodiments, pleural cancer includes, but is not limited to lung cancer, metastases, mesothelioma, malignant mesothelioma, lymphoma, malignant fibrous tumor, sarcoma, askin tumor, extraskeletal osteosarcoma, malignant fibrous histiocytoma, solitary fibrous tumor, lipoma, mesothelial cyst, calcifying fibrous pseudotumor, primary effusion lymphoma and any other hyperproliferative disease, besides neoplasia, located in an organ system listed above.

[0059] In some embodiments, the cancer is mesothelioma. In some embodiments, the mesothelioma is a pleural mesothelioma, peritoneal mesothelioma, pericardial mesothelioma, testicular mesothelioma, epithelioid mesothelioma, sarcomatoid mesothelioma, biphasic mesothelioma, small cell mesothelioma, deciduoid mesothelioma, cystic and papillary mesothelioma, desmoplastic mesothelioma, adenomatoid mesothelioma, heterologous mesothelioma, well-defined papillary cell mesothelioma, or any combination thereof. In some embodiments, the mesothelioma is a pleural mesothelioma. In some embodiments, the mesothelioma is a peritoneal mesothelioma. In some embodiments, the mesothelioma is a pericardial mesothelioma. In some embodiments, the mesothelioma is a testicular mesothelioma. In some embodiments, the mesothelioma is a epithelioid mesothelioma. In some embodiments, the mesothelioma is a sarcomatoid mesothelioma. In some embodiments, the 20 4865-0355-2478, v. 1mesothelioma is a biphasic mesothelioma. In some embodiments, the mesothelioma is a small cell mesothelioma. In some embodiments, the mesothelioma is a deciduoid mesothelioma. In some embodiments, the mesothelioma is a cystic and papillary mesothelioma. In some embodiments, the mesothelioma is a desmoplastic mesothelioma. In some embodiments, the mesothelioma is a adenomatoid mesothelioma. In some embodiments, the mesothelioma is a heterologous mesothelioma. In some embodiments, the mesothelioma is a well-defined papillary cell mesothelioma.

[0060] In some embodiments, the disease is an autoimmune disease. An autoimmune disease a disease in which the body’s own immune system mistakenly attacks its own healthy cells and tissues. In one embodiment, the disease is multiple sclerosis. In one embodiment, the disease is Crohn’s disease. In one embodiment, the disease is ulcerative colitis. V. Definitions

[0061] The use of the word “a” or “an”, when used in conjunction with the term “comprising” in the claims and / or the specification, may mean “one,” but it is also consistent with the meaning of “one or more,” “at least one,” and “one or more than one.” As used herein “another” may mean at least a second or more.

[0062] As used herein, the terms “active pharmaceutical ingredient”, “drug”, “pharmaceutical”, “active agent”, “therapeutic agent”, and “therapeutically active agent” are used interchangeably to represent a compound which invokes a therapeutic or pharmacological effect in a human or animal and is used to treat a disease, disorder, or other condition. In some embodiments, these compounds have undergone and received regulatory approval for administration to a living creature.

[0063] The use of the term “or” in the claims is used to mean “and / or” unless explicitly indicated to refer to alternatives only or the alternatives are mutually exclusive. As used herein “another” may mean at least a second or more.

[0064] The terms “compositions,” “pharmaceutical compositions,” “formulations,” “pharmaceutical formulations,” “preparations”, and “pharmaceutical preparations” are used synonymously and interchangeably herein.

[0065] “Treating” or treatment of a disease or condition refers to executing a protocol, which may include administering one or more drugs to a patient, to alleviate signs or symptoms of the disease. Desirable effects of treatment include decreasing the rate of disease progression, 21 4865-0355-2478, v. 1ameliorating or palliating the disease state, and remission or improved prognosis. Alleviation can occur before signs or symptoms of the disease or condition appearing, as well as after their appearance. Thus, “treating” or “treatment” may include “preventing” or “prevention” of disease or undesirable condition. In addition, “treating” or “treatment” does not require complete alleviation of signs or symptoms, does not require a cure, and specifically includes protocols that have only a marginal effect on the patient.

[0066] The term “therapeutic benefit” or “therapeutically effective” as used throughout this application refers to anything that promotes or enhances the well-being of the subject with respect to the medical treatment of this condition. This includes, but is not limited to, a reduction in the frequency or severity of the signs or symptoms of a disease. For example, treatment of cancer may involve, for example, a reduction in the size of a tumor, a reduction in the invasiveness of a tumor, a reduction in the growth rate of the cancer, or prevention of metastasis. Treatment of cancer may also refer to prolonging the survival of a subject with cancer.

[0067] “Subject” and “patient” refer to either a human or non-human, such as primates, mammals, and vertebrates. In particular embodiments, the subject is a human.

[0068] As generally used herein “pharmaceutically acceptable” refers to those compounds, materials, compositions, and / or dosage forms which are, within the scope of sound medical judgment, suitable for use in contact with the tissues, organs, and / or bodily fluids of human beings and animals without excessive toxicity, irritation, allergic response, or other problems or complications commensurate with a reasonable benefit / risk ratio.

[0069] “Pharmaceutically acceptable salts” means salts of compounds disclosed herein which are pharmaceutically acceptable, as defined above, and which possess the desired pharmacological activity. Such salts include acid addition salts formed with inorganic acids such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, and the like; or with organic acids such as 1,2-ethanedisulfonic acid, 2-hydroxyethanesulfonic acid, 2-naphthalenesulfonic acid, 3-phenylpropionic acid, 4,4′-methylenebis(3-hydroxy-2-ene- 1-carboxylic acid), 4-methylbicyclo[2.2.2]oct-2-ene-1-carboxylic acid, acetic acid, aliphatic mono- and dicarboxylic acids, aliphatic sulfuric acids, aromatic sulfuric acids, benzenesulfonic acid, benzoic acid, camphorsulfonic acid, carbonic acid, cinnamic acid, citric acid, cyclopentanepropionic acid, ethanesulfonic acid, fumaric acid, glucoheptonic acid, gluconic acid, glutamic acid, glycolic acid, heptanoic acid, hexanoic acid, hydroxynaphthoic acid, lactic 22 4865-0355-2478, v. 1acid, laurylsulfuric acid, maleic acid, malic acid, malonic acid, mandelic acid, methanesulfonic acid, muconic acid, o-(4-hydroxybenzoyl)benzoic acid, oxalic acid, p-chlorobenzenesulfonic acid, phenyl-substituted alkanoic acids, propionic acid, p-toluenesulfonic acid, pyruvic acid, salicylic acid, stearic acid, succinic acid, tartaric acid, tertiarybutylacetic acid, trimethylacetic acid, and the like. Pharmaceutically acceptable salts also include base addition salts which may be formed when acidic protons present are capable of reacting with inorganic or organic bases. Acceptable inorganic bases include sodium hydroxide, sodium carbonate, potassium hydroxide, aluminum hydroxide, and calcium hydroxide. Acceptable organic bases include ethanolamine, diethanolamine, triethanolamine, tromethamine, N-methylglucamine, and the like. It should be recognized that the particular anion or cation forming a part of any salt of this invention is not critical, so long as the salt, as a whole, is pharmacologically acceptable. Additional examples of pharmaceutically acceptable salts and their methods of preparation and use are presented in Handbook of Pharmaceutical Salts: Properties, and Use (P. H. Stahl & C. G. Wermuth eds., Verlag Helvetica Chimica Acta, 2002).

[0070] The term “derivative thereof” refers to any chemically modified compound, wherein at least one of the compounds is modified by substitution of atoms or molecular groups or bonds. In one embodiment, a derivative thereof is a salt thereof. Salts are, for example, salts with suitable mineral acids, such as hydrohalic acids, sulfuric acid or phosphoric acid, for example, hydrochlorides, hydrobromides, sulfates, hydrogen sulfates or phosphates, salts with suitable carboxylic acids, such as optionally hydroxylated lower alkanoic acids, for example, acetic acid, glycolic acid, propionic acid, lactic acid or pivalic acid, optionally hydroxylated and / or oxo-substituted lower alkane dicarboxylic acids, for example, oxalic acid, succinic acid, fumaric acid, maleic acid, tartaric acid, citric acid, pyruvic acid, malic acid, ascorbic acid, and also with aromatic, heteroaromatic or araliphatic carboxylic acids, such as benzoic acid, nicotinic acid or mandelic acid, and salts with suitable aliphatic or aromatic sulfonic acids or N-substituted sulfamic acids, for example, methanesulfonates, benzenesulfonates, p-toluenesulfonates or N-cyclohexylsulfamates (cyclamates).

[0071] As used in this specification and claim(s), the words “comprising” (and any form of comprising, such as “comprise” and “comprises”), “having” (and any form of having, such as “have” and “has”), “including” (and any form of including, such as “includes” and “include”), or “containing” (and any form of containing, such as “contains” and “contain”) are inclusive or open-ended and do not exclude additional, unrecited elements or method steps. 23 4865-0355-2478, v. 1

[0072] As used in this specification, the term “significant” (and any form of significance such as “significantly”) is not meant to imply statistical differences between two values but only to imply importance or the scope of the difference of the parameter.

[0073] Throughout this application, the term “about” is used to indicate that a value includes the inherent variation of error for the device, the method being employed to determine the value or the variation that exists among the study subjects or experimental studies. Unless another definition is applicable, the term “about” refers to ±10% of the indicated value.

[0074] As used herein, the term “substantially free of” or “substantially free” in terms of a specified component, is used herein to mean that none of the specified components has been purposefully formulated into a composition and / or is present only as a contaminant or in trace amounts. The total amount of all containments, by-products, and other material is present in that composition in an amount of less than 2%. The term “essentially free of” or “essentially free” is used to represent that the composition contains less than 1% of the specific component. The term “entirely free of” or “entirely free” contains less than 0.1% of the specific component.

[0075] The term “homogenous” is used to mean a composition in which the components are mixed in such a way that the components are uniformly distributed amongst the composition. In a preferred embodiment, the composition is uniformly distributed in such a manner that there are no regions of a single component that are greater than 1 µm or more preferably less than 0.1 µm. In one embodiment, the composition is so homogeneously mixed in such a manner that there are no atoms of the thermally conductive excipient are adjacent to another atom of the thermally conductive excipient.

[0076] The terms “substantially” or “approximately” as used herein may be applied to modify any quantitative comparison, value, measurement, or other representation that could permissibly vary without resulting in a change in the basic function to which it is related. Such an amount, when referring to the amount of the composition that is in the liquid phase, refers to an amount of composition such that the compostion appears liquid or soft to the naked eye.

[0077] A temperature, when used without any other modifier, refers to room temperature, preferably 23 °C unless otherwise noted. An elevated temperature is a temperature that is more than 5 °C greater than room temperature; preferably more than 10 °C greater than room temperature.

[0078] The term “unit dose” refers to a formulation of the pharmaceutical composition such that the formulation is prepared in a manner sufficient to provide a single therapeutically 24 4865-0355-2478, v. 1effective dose of the active pharmaceutical ingredient to a patient in a single administration. Such unit dose formulations that may be used include but are not limited to a single tablet, capsule, or other oral formulations, or a single vial with a syringeable liquid or other injectable formulations. The resulting product can then undergo further downstream processing to create an intermediate product, such as granules, that can then be further formulated into a unit dose such as one prepared for oral delivery as tablets, capsules, three-dimensionally printed selective laser sintered (3DPSLS) or suspensions; pulmonary and nasal delivery; topical delivery as emulsions, ointments or creams; transdermal delivery; and parenteral delivery as suspensions, microemulsions or depot. In some forms, the final pharmaceutical composition that is produced is no longer a powder and is further produced as a homogenous final product. This final product has the capability of being processed into granules and being compressed or 3DPSLS into a final pharmaceutical unit dose form.

[0079] Notwithstanding that the numerical ranges and parameters setting forth the broad scope of the invention are approximations, the numerical values set forth in the specific examples are reported as precisely as possible. Any numerical value, however, inherently contains certain errors necessarily resulting from the standard deviation found in their respective testing measurements and parameters.

[0080] Other objects, features, and advantages of the present disclosure will become apparent from the following detailed description. It should be understood, however, that the detailed description and the specific examples, while indicating preferred embodiments of the disclosure, are given by way of illustration only, since various changes and modifications within the spirit and scope of the disclosure will become apparent to those skilled in the art from this detailed description. VI. Examples

[0081] To facilitate a better understanding of the present disclosure, the following examples of specific embodiments are given. It should be appreciated by those of skill in the art that the techniques disclosed in the examples which follow represent techniques discovered by the inventor to function well in the practice of the disclosure, and thus can be considered to constitute preferred modes for its practice. However, those of skill in the art should, in light of the present disclosure, appreciate that many changes can be made in the specific embodiments which are disclosed and still obtain a like or similar result without departing from the spirit and 25 4865-0355-2478, v. 1scope of the disclosure. In no way should the following examples be read to limit or define the entire scope of the disclosure. Example 1 – Preparation of Formulations with HME and Dielectric Heating In Parallel Table 1: Formulation composition Formulation 1 Ingredient Purpose % weight

[0082] First, the biologic active pharmaceutical ingredient, bovine serum albumin was physically mixed with Kolliphor P 407®. This physical mixture was then fed into the SLS printer powder bed and subjected to sintering to form consolidated structures. Table 2: Printing parameters: Printing parameters Surface temperature (°C) 43the attached FIGS.1-4. * * *

[0084] All of the compositions and methods disclosed and claimed herein can be made and executed without undue experimentation in light of the present disclosure. While the compositions and methods of this disclosure have been described in terms of preferred embodiments, it will be apparent to those of skill in the art that variations may be applied to the methods and in the steps or in the sequence of steps of the method described herein without departing from the concept, spirit, and scope of the disclosure. More specifically, it will be apparent that certain agents which are both chemically and physiologically related may be substituted for the agents described herein while the same or similar results would be achieved. All such similar substitutes and modifications apparent to those skilled in the art are deemed to be within the spirit, scope, and concept of the disclosure as defined by the appended claims. 26 4865-0355-2478, v. 1References

[0001] The following references, to the extent that they provide exemplary procedural or other details supplementary to those set forth herein, are specifically incorporated herein by reference. Chen et al., Adv. Drug Deliv. Rev.172:211–233, 2021. Davis et al., J. Pharm. Sci.110:1432–1443, 2021. Eibl et al., Appl Microbiol Biotechnol., 86(1):41–9, 2010. Fina et al., Int. J. Pharm.529:285–293, 2017. Geraldes et al., J. Drug Target.28:339–355, 2020. Hill, BCM Med., 10(10):1–3, 2012. Karri et al., J. Surg. Res.190:655–661, 2014. Kruth et al., Rapid Prototyping Journal, 11(1): 26–36, 2005. Mulcahy et al., Rand Health Q., 7(4):3, 2018. Narasimhan et al., Annu. Rev. Biomed. Eng.18:25–49, 2016. Reichert et al., J Membr Sci, 199:161–6, 2002. Rose et al., Humana Press; 2003. Sarpatwari et al., N Engl J Med., 372(25):2380–2, 2015. Schmidt et al., Powder Technology, 228:84–90. Shimoni et al., Bioprocess Int, 11(10):26–31, 2013. Thakkar et al., Eur. J. Pharm. Biopharm.163:141–156, 2021. Yang et al., Rapid Prototyping Journal, 24(6): 1059–1066, 2018.

Claims

What Is Claimed Is:

1. A method of preparing a pharmaceutical composition comprising: (A) obtaining a composition comprising: (i) an amino acid based biologic active pharmaceutical ingredient; and (ii) a polymer, wherein the polymer is in the semi-crystalline or crystalline state; (B) applying the composition to a surface of a chamber; wherein the chamber has been set to a first temperature; and (C) melting the composition with a laser to obtain the pharmaceutical composition.

2. The method of claim 1, wherein the amino acid based biologic active pharmaceutical ingredient is a peptide.

3. The method of claim 1, wherein the amino acid based biologic active pharmaceutical ingredient is a protein.

4. The method of claim 1, wherein the amino acid based biologic active pharmaceutical ingredient is an antibody.

5. The method according to any one of claims 1-4, wherein the composition comprises an amount from about 1% w / w to about 90% w / w of the amino acid based biologic active pharmaceutical ingredient.

6. The method of claim 5, wherein the amount of the amino acid based biologic active pharmaceutical ingredient is from about 2.5% w / w to about 50% w / w of the composition.

7. The method of either claim 5 or claim 6, wherein the amount of the amino acid based biologic active pharmaceutical ingredient is from about 5% w / w to about 25% w / w of the composition.

8. The method according to any one of claims 5-7, wherein the amount of the amino acid based biologic active pharmaceutical ingredient is from about 5% w / w to about 25% w / w of the composition.

9. The method according to any one of claims 1-8, wherein the composition comprises an amount from about 10% w / w to about 99% w / w of the polymer.

10. The method of claim 9, wherein the amount of the polymer is from about 50% w / w to about 97.5% w / w of the composition.

11. The method of either claim 9 or claim 10, wherein the amount of the polymer is from about 75% w / w to about 95% w / w of the composition. 28 4865-0355-2478, v.

112. The method according to any one of claims 9-11, wherein the amount of the polymer is from about 85% w / w to about 95% w / w of the composition.

13. The method according to any one of claims 1-12, wherein the polymer is in a semi- crystalline state.

14. The method according to any one of claims 1-12, wherein the polymer is in a crystalline state.

15. The method according to any one of claims 1-14, wherein the polymer comprises one or more blocks of polyethylene oxide or polypropylene oxide.

16. The method according to any one of claims 1-15, wherein the polymer comprises one or more polyethylene oxide blocks.

17. The method according to any one of claims 1-16, wherein the polymer is a diblock or a triblock copolymer.

18. The method of claim 17, wherein the polymer is a triblock copolymer.

19. The method of claim 18, wherein the triblock copolymer comprises two polyethylene glycol blocks.

20. The method of either claim 18 or claim 19, wherein the triblock copolymer comprises a polypropylene glycol block.

21. The method according to any one of claims 18-20, wherein the triblock copolymer comprises two polyethylene glycol blocks on either side of a polypropylene glycol block.

22. The method according to any one of claims 18-21, wherein the triblock copolymer comprises one or more polyethylene glycol block with a number of repeating units from 60 to about 140.

23. The method of claim 22, wherein the number of repeating units in the polyethylene glycol block is from about 80 to about 120.

24. The method according to any one of claims 18-23, wherein the triblock copolymer comprises a polypropylene glycol block with a number of repeating units from about 30 to about 80 repeating units.

25. The method of claim 24, wherein the number of repeating units in the polypropylene glycol block is from about 50 to about 60.

26. The method according to any one of claims 1-25, wherein the polymer is a triblock copolymer comprising two polyethylene glycol with a number of repeating units from about 80 to about 120 and a polypropylene glycol with a number of repeating units from about 50 to about 60. 29 4865-0355-2478, v.

127. The method according to any one of claims 1-26, wherein the laser is a CO2laser.

28. The method according to any one of claims 1-27, wherein the method comprises using a laser with a speed from about 25 mm / s to about 300 mm / s.

29. The method of claim 28, wherein the speed is from about 50 mm / s to about 250 mm / s.

30. The method of either claim 28 or claim 29, wherein the speed is from about 100 mm / s to about 200 mm / s.

31. The method according to any one of claims 1-30, wherein the method comprises a hatch spacing between the laser from about 10 µm to about 1 mm.

32. The method of claim 31, wherein the hatch spacing is from about 50 µm to 500 µm.

33. The method of either claim 31 or claim 32, wherein the hatch spacing is from about 100 µm to 300 µm.

34. The method according to any one of claims 1-27, wherein the first temperature is below 100 °C.

35. The method according to any one of claims 1-34, wherein the first temperature is from about −120 °C to about 80 °C.

36. The method of claim 35, wherein the first temperature is from about −100 °C to about 60 °C.

37. The method of either claim 35 or claim 36, wherein the first temperature is from about 0 °C to about 50 °C.

38. The method according to any one of claims 1-37, wherein the method comprises adjusting the temperature of the surface to a surface temperature that is distinct from the first temperature.

39. The method according to any one of claims 1-34, wherein the surface temperature is from about −120 °C to about 80 °C.

40. The method of claim 35, wherein the surface temperature is from about −100 °C to about 60 °C.

41. The method of either claim 39 or claim 40, wherein the surface temperature is from about 0 °C to about 50 °C.

42. The method according to any one of claims 38-41, wherein the difference between the first temperature and the surface temperature is from about 0.1 °C to about 10 °C.

43. The method of claim 42, wherein the difference is from about 0.5 °C to about 5 °C.

44. The method according to any one of claims 1-43, wherein the laser is used in an additive manufacturing technique. 30 4865-0355-2478, v.

145. The method of claim 44, wherein the additive manufacturing technique is a melting process.

46. The method of either claim 44 or claim 45, wherein the additive manufacturing technique is powder bed fusion.

47. The method according to any one of claims 1-46, wherein the method further comprises one or more processing steps.

48. The method of claim 47, wherein the processing step is blending.

49. The method of claim 47, wherein the processing step is milling.

50. The method of claim 47, wherein the processing step is tablet pressing.

51. The method of claim 47, wherein the processing step is coating.

52. A pharmaceutical composition comprising (A) an amino acid based biologic active pharmaceutical ingredient; and (B) a pharmaceutically acceptable polymer, wherein the pharmaceutically acceptable polymer comprises at least 80% of the composition and the pharmaceutically acceptable polymer is in the semi-crystalline or crystalline state.

53. The pharmaceutical composition of claim 52, wherein the amino acid based biologic active pharmaceutical ingredient is a peptide.

54. The pharmaceutical composition of claim 52, wherein the amino acid based biologic active pharmaceutical ingredient is protein.

55. The pharmaceutical composition of claim 52, wherein the amino acid based biologic active pharmaceutical ingredient is an antibody.

56. The pharmaceutical composition according to any one of claims 52-55, wherein the composition comprises an amount from about 1% w / w to about 20% w / w of the amino acid based biologic active pharmaceutical ingredient.

57. The pharmaceutical composition of claim 56, wherein the amount of the amino acid based biologic active pharmaceutical ingredient is from about 2.5% w / w to about 17.5% w / w of the composition.

58. The method of either claim 56 or claim 57, wherein the amount of the amino acid based biologic active pharmaceutical ingredient is from about 5% w / w to about 15% w / w of the composition.

59. The pharmaceutical composition according to any one of claims 56-58, wherein the amount of the amino acid based biologic active pharmaceutical ingredient is from about 7.5% w / w to about 12.5% w / w of the composition. 31 4865-0355-2478, v.

160. The pharmaceutical composition according to any one of claims 52-59, wherein the composition comprises an amount from about 80% w / w to about 99% w / w of the polymer.

61. The pharmaceutical composition of claim 60, wherein the amount of the polymer is from about 82.5% w / w to about 97.5% w / w of the composition.

62. The pharmaceutical composition of either claim 60 or claim 61, wherein the amount of the polymer is from about 85% w / w to about 95% w / w of the composition.

63. The pharmaceutical composition according to any one of claims 60-62, wherein the amount of the polymer is from about 87.5% w / w to about 95% w / w of the composition.

64. The pharmaceutical composition according to any one of claims 52-63, wherein the polymer is in a semi-crystalline state.

65. The pharmaceutical composition according to any one of claims 52-63, wherein the polymer is in a crystalline state.

66. The pharmaceutical composition according to any one of claims 52-65, wherein the polymer comprises one or more blocks of polyethylene oxide or polypropylene oxide.

67. The pharmaceutical composition according to any one of claims 52-66, wherein the polymer comprises one or more polyethylene oxide blocks.

68. The pharmaceutical composition according to any one of claims 52-67, wherein the polymer is a diblock or a triblock copolymer.

69. The pharmaceutical composition of claim 68, wherein the polymer is a triblock copolymer.

70. The pharmaceutical composition of claim 69, wherein the triblock copolymer comprises two polyethylene glycol blocks.

71. The pharmaceutical composition of either claim 69 or claim 70, wherein the triblock copolymer comprises a polypropylene glycol block.

72. The pharmaceutical composition according to any one of claims 69-71, wherein the triblock copolymer comprises two polyethylene glycol blocks on either side of a polypropylene glycol block.

73. The pharmaceutical composition according to any one of claims 69-72, wherein the triblock copolymer comprises one or more polyethylene glycol block with a number of repeating units from 60 to about 140.

74. The pharmaceutical composition of claim 73, wherein the number of repeating units in the polyethylene glycol block is from about 80 to about 120. 32 4865-0355-2478, v.

175. The pharmaceutical composition according to any one of claims 69-74, wherein the triblock copolymer comprises a polypropylene glycol block with a number of repeating units from about 30 to about 80 repeating units.

76. The pharmaceutical composition of claim 75, wherein the number of repeating units in the polypropylene glycol block is from about 50 to about 60.

77. The pharmaceutical composition according to any one of claims 52-76, wherein the polymer is a triblock copolymer comprising two polyethylene glycol with a number of repeating units from about 80 to about 120 and a polypropylene glycol with a number of repeating units from about 50 to about 60.

78. A method of treating or preventing a disease or disorder comprising administering a pharmaceutical composition according to any one of claims 52-77 to a patient in need thereof, wherein the pharmaceutical composition comprises an amino acid based active pharmaceutical ingredient capable of treating or preventing the disease or disorder. 33 4865-0355-2478, v. 1

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