Polypeptide formulations for oral delivery

Abstract

The present disclosure provides, among other things, methods and compositions for the oral administration of semaglutide polypeptides. The present disclosure provides methods and compositions that include a semaglutide polypeptide formulation, e.g., tablet formulation suitable for oral administration, where the formulation includes a crystallized polypeptide composition and a pharmaceutically acceptable carrier.

Description

POLYPEPTIDE FORMULATIONS FOR ORAL DELIVERYCROSS-REFERENCE TO RELATED APPLICATION

[0001] This application claims priority to Indian Application No. 202441030520, filed April 16, 2024, the entirety of each of which is incorporated herein by reference.BACKGROUND

[0002] Semaglutide is an important therapeutic, approved under the trade names Ozempic®, Rybelsus®, and Wegovy®, and, as adjuncts to diet and exercize to approve glycemic control for indications relating to weight management, glycemic control and / or risk of adverse cardiovascular events, in some cases in adults with type 2 diabetes mellitus, and in others in obese individuals. Ozempic®, and Wegovy® are administered by injection;Rybelsus® is an oral tablet.SUMMARY

[0003] The present disclosure provides, among other things, technologies (e.g., methods and compositions) for the oral administration of crystalline semaglutide, and documents surprising and unexpected properties of provided such technologies. Thus, the present disclosure provides technologies (e.g., methods and compositions) for the administration (e.g., oral administration) of crystalline semaglutide polypeptide compositions.

[0004] Among other things, the present disclosure identifies the source of a problem with conventional oral semaglutide formulations, such as, for example, inefficient delivery of semaglutide to the bloodstream.

[0005] The present disclosure provides an insight that technologies which achieve rapid oral delivery of a semaglutide are particularly valuable. The present disclosure provides the surprising finding that certain formulations of semaglutide polypeptides, e.g., specifically formulations of crystalline semaglutide polypeptides, can achieve rapid delivery of semaglutide polypeptides via oral administration of the formulations.

[0006] Among other things, the present disclosure provides a surprising finding that certain formulations (.e.g, certain tablet formulations) of semaglutide polypeptides, e.g.,specifically formulations of crystalline semaglutide polypeptides, can achieve particularly rapid delivery of semaglutide polypeptides via oral administration of the formulations.

[0007] The present disclosure further provides a surprising finding that certain formulations (e.g., certain tablet formulations) of semaglutide polypeptides, e.g., specifically formulations of crystalline semaglutide polypeptides, wherein the tablet includes (e.g., is coated with) Vitamin E, can achieve particularly rapid delivery of semaglutide polypeptides via oral administration of the formulations.

[0008] The present disclosure further provides a surprising finding that certain formulations of semaglutide polypeptides, e.g., specifically formulations of crystalline semaglutide polypeptides, can achieve effective systemic delivery of semaglutide polypeptides via oral administration of the formulations.

[0009] Thus, the present disclosure provides, among other things, semaglutide polypeptide formulations (e.g., tablets) for oral delivery. In many embodiments, provided formulations comprise or otherwise utilize a semaglutide polypeptide in a crystalline (i.e., crystallized) form. Alternatively or additionally, in many embodiments, provided formulations include (e.g., are coated with) Vitamin E.

[0010] In some embodiments, the present disclosure provides formulations that comprise a carrier polypeptide. In many such embodiments, the carrier polypeptide is crystallized. In some embodiments, a crystalline semaglutide polypeptide composition is a composition in which semaglutide polypeptide is dispersed within such crystalline carrier polypeptide. Thus, in some embodiments, the present disclosure encompasses an insight that during a process of crystallizing a carrier polypeptide composition, a semaglutide polypeptide composition can be embedded in such carrier polypeptide composition, thereby creating a composition comprising both a carrier polypeptide composition and a semaglutide polypeptide composition. In some embodiments, carrier polypeptide and semaglutide polypeptide form a co-crystal during crystallization. In some embodiments, such composition comprising both a carrier polypeptide composition and a semaglutide polypeptide composition (e.g., a co-crystal of carrier polypeptide composition and a polypeptide composition) is amenable for formulation.

[0011] Those skilled in the art, reading the present disclosure, will appreciate that teachings provided herein significantly improve efficiency of developing semaglutide polypeptide formulations (e.g., beyond explicitly exemplified formulations).

[0012] In some embodiments, the present disclosure provides oral formulations (e.g., tablet) that comprise (1) a semaglutide polypeptide composition (e.g., a crystalline semaglutide polypeptide composition which may, in some embodiments, be a powder); and (2) one or more pharmaceutically acceptable carriers or excipients. In some embodiments, provided oral formulations include (3) a carrier polypeptide composition (e.g., a crystalline carrier polypeptide composition).

[0013] In some embodiments of those formulations that include a carrier polypeptide composition, such carrier polypeptide composition co-exists with a semaglutide polypeptide composition. In some embodiments, a semaglutide polypeptide composition is embedded in a carrier polypeptide composition. In some embodiments, a carrier polypeptide composition is embedded in a semaglutide polypeptide composition.

[0014] In some embodiments of those formulations that include a carrier polypeptide composition, such carrier polypeptide composition is an amorphous composition and a semaglutide polypeptide composition is a crystallized composition. In some embodiments, a carrier polypeptide composition is a crystallized composition and a semaglutide polypeptide composition is crystallized composition. In some embodiments, a carrier polypeptide composition and a semaglutide polypeptide composition form a co-crystal.

[0015] In many embodiments, a carrier polypeptide composition and a semaglutide polypeptide composition of provided formulations each independently are or comprise a polypeptide in a non-liquid phase - e.g., such formulations may be tablets, gels, solids, suspensions (e.g., of solid particles), etc.

[0016] In some embodiments, a semaglutide polypeptide formulation is or comprises a truffle formulation. In some embodiments, a truffle formulation comprises a pharmaceutically acceptable shell and a core that comprises (1) a semaglutide polypeptide composition (e.g., a crystalline semaglutide polypeptide composition which may, in some embodiments, be a powder); and (2) one or more pharmaceutically acceptable carriers or excipients. In some embodiments, provided oral formulations include (3) a carrier polypeptide composition (e.g., a crystalline carrier polypeptide composition).

[0017] In some embodiments, a semaglutide polypeptide formulation is or comprises a tablet formulation that comprises (1) a semaglutide polypeptide composition (e.g., a crystalline semaglutide polypeptide composition which may, in some embodiments, be a powder); and (2) one or more pharmaceutically acceptable carriers or excipients. In someembodiments, provided oral formulations include (3) a carrier polypeptide composition (e.g., a crystalline carrier polypeptide composition).

[0018] In some embodiments, a semaglutide polypeptide formulation is or comprises a globule formulation that comprises (1) a semaglutide polypeptide composition (e.g., a crystalline semaglutide polypeptide composition which may, in some embodiments, be a powder); and (2) one or more pharmaceutically acceptable carriers or excipients. In some embodiments, provided oral formulations include (3) a carrier polypeptide composition (e.g., a crystalline carrier polypeptide composition).

[0019] In some embodiments, a semaglutide polypeptide formulation is or comprises a candy formulation that comprises (1) a semaglutide polypeptide composition (e.g., a crystalline semaglutide polypeptide composition which may, in some embodiments, be a powder); and (2) one or more pharmaceutically acceptable carriers or excipients. In some embodiments, provided oral formulations include (3) a carrier polypeptide composition (e.g., a crystalline carrier polypeptide composition).

[0020] In some embodiments, a semaglutide polypeptide formulation is or comprises a capsule formulation that comprises (1) a semaglutide polypeptide composition (e.g., a crystalline semaglutide polypeptide composition which may, in some embodiments, be a powder); and (2) one or more pharmaceutically acceptable carriers or excipients. In some embodiments, provided oral formulations include (3) a carrier polypeptide composition (e.g., a crystalline carrier polypeptide composition). .

[0021] In some embodiments, a provided oral composition, whether formulated, for example, as a tablet or capsule, or to be or include a truffle or globule or candy, includes a Vitamin E agent (e.g., Vitamin E). In some embodiments, a Vitamin E agent may be considered to be a pharmaceutically acceptable carrier. In some particular embodiments, a provided composition (e.g., a provided pharmaceutical composition) is coated with a Vitamin E agent. In certain specific embodiments, a provided tablet composition comprises (e.g., is coated with) a Vitamin E agent; in certain specific embodiment, a provided capsule formulation comprises a Vitamin E agent.

[0022] The present disclosure includes the surprising discovery that oral administration of formulations disclosed herein, e.g., of truffle, tablet, globule, candy, and / or capsule formulations as described herein can successfully deliver semaglutide polypeptides to the bloodstream with particularly desirable characteristics. The present disclosure furtherincludes the surprising discovery that semaglutide polypeptides orally administered in a formulation as disclosed herein e.g., a tablet formulation, a truffle formulation, a globule formulation, a candy formulation, a capsule formulation are particularly efficiently delivered to the bloodstream, lymphatic system, and thoracic duct.

[0023] Advantages of formulations provided herein include that they can effectively deliver polypeptides (e.g., of therapeutic semaglutide polypeptides) to the bloodstream after oral administration of such formulations. Provided technologies have broad applicability and offer a new platform for semaglutide polypeptide formulation. Upon reading the present disclosure, those skilled in the art will appreciate its generality and applicability to semaglutide polypeptides of interest.

[0024] The present disclosure demonstrates unexpectedly advantageous characteristics of provided formulations, including, in some embodiments, effectiveness in one or more of various modes of oral administration. Indeed, the present disclosure documents that provided formulations can achieve desirable pharmacokinetic properties, including, for example, one or more of rate of delivery into the bloodstream, increased halflife, etc., and in some embodiments can achieve such properties via one or more of such oral administration modes.

[0025] In some embodiments, the present disclosure provides polypeptide formulations for oral delivery, which formulations include (1) a semaglutide polypeptide composition (e.g., a crystalline semaglutide polypeptide composition which may, in some embodiments, be a powder); and (2) one or more pharmaceutically acceptable carriers or excipients, and, optionally (3) a carrier polypeptide composition (e.g., a crystalline carrier polypeptide composition) , optionally wherein the formulation is a truffle formulation comprising a core and a pharmaceutically acceptable shell, wherein the core comprises a semaglutide polypeptide composition (e.g., a crystalline polypeptide composition which, in some embodiments, may be in powder form).

[0026] In some embodiments, the present disclosure provides polypeptide formulations for oral delivery that are tablet semaglutide polypeptide formulations, which tablet semaglutide polypeptide formulation comprises 1) a semaglutide polypeptide composition (e.g., a crystalline semaglutide polypeptide composition which may, in some embodiments, be a powder); and (2) one or more pharmaceutically acceptable carriers orexcipients, and, optionally (3) a carrier polypeptide composition (e.g., a crystalline carrier polypeptide composition).

[0027] In some embodiments a polypeptide formulation for oral delivery is a tablet (e.g., tablet formulation).

[0028] In some embodiments, the present disclosure provides polypeptide formulations for oral delivery, which formulations are globule formulations that comprise (1) a semaglutide polypeptide composition (e.g., a crystalline semaglutide polypeptide composition which may, in some embodiments, be a powder); and (2) one or more pharmaceutically acceptable carriers or excipients, and, optionally (3) a carrier polypeptide composition (e.g., a crystalline carrier polypeptide composition).

[0029] In some embodiments, the present disclosure provides polypeptide formulations for oral delivery, which formulations are candy formulations that comprise 1) a semaglutide polypeptide composition (e.g., a crystalline semaglutide polypeptide composition which may, in some embodiments, be a powder); and (2) one or more pharmaceutically acceptable carriers or excipients, and, optionally (3) a carrier polypeptide composition (e.g., a crystalline carrier polypeptide composition) , optionally wherein the candy formulation is in a form, for example, of candy gems, chewing gum, gummy candy, hard candy (e.g., drops, lollipops, lozenges, rock candy, stick candy, etc.) marshmallows, syrup, toffee, etc.. In some embodiments, a candy formulation may be in the form of a drop, film, gel, patch, spray, wafer, etc.

[0030] In some embodiments, a provided formulation may be in the form of a dry powder (e.g., a dry spray).

[0031] In some embodiments, a provided formulation may be in the form of a fastdissolving tablet.

[0032] In some embodiments, a provided formulation may be in the form of a fastdissolving film.

[0033] In some embodiments, the present disclosure provides polypeptide formulations for oral delivery, which formulations are capsule formulations that comprise (1) a semaglutide polypeptide composition (e.g., a crystalline semaglutide polypeptide composition which may, in some embodiments, be a powder); (2) one or more pharmaceutically acceptable carriers or excipients; optionally (3) a carrier polypeptidecomposition (e.g., a crystalline carrier polypeptide composition) , optionally wherein a pharmaceutical acceptable carrier comprises a Vitamin E agent.

[0034] In certain embodiments, a provided semaglutide polypeptide formulation comprises about 1 mg to about 2,000 mg of a Vitamin E agent (e.g., when a total formulation is about 5 mg to about 200 g), optionally where a provided semaglutide polypeptide formulation comprises about 1 mg to about 1,000 mg, about 1 mg to about 500 mg, about 1 mg to about 400 mg, about 1 mg to about 300 mg, about 1 mg to about 200 mg, about 1 mg to about 100 mg, about 1 mg to about 50mg, or about 1 mg to about 25 mg of a Vitamin E agent.

[0035] In certain embodiments, a provided semaglutide polypeptide formulation comprises a molar excess or excess by weight of a Vitamin E agent relative to the amount of active semaglutide polypeptide in the semaglutide polypeptide formulation, optionally where in the excess is a fold excess of at least 1.1, 1.5, 2, 3, 4, 5, 6, 7, 8, 9, 10, 20, 30, 40, 50, 60, 70, 80, 90, 100, 500, or 1,000 fold.

[0036] In some embodiments, the present disclosure provides methods of delivering a semaglutide polypeptide to the bloodstream of a subject, which methods comprise orally administering a semaglutide polypeptide composition as described herein. For example, the present disclosure provides methods comprising a step of orally administering to a subject in need thereof a semaglutide polypeptide formulation comprising 1) a semaglutide polypeptide composition (e.g., a crystalline semaglutide polypeptide composition which may, in some embodiments, be a powder); (2) one or more pharmaceutically acceptable carriers or excipients; optionally (3) a carrier polypeptide composition (e.g., a crystalline carrier polypeptide composition) , optionally wherein a pharmaceutical acceptable carrier comprises a Vitamin E agent. . In some embodiments, such an administered formulation is or comprises a truffle formulation. Alternatively or additionally, in some embodiments, an administered formulation is or comprises a globule formulation, and / or an administered formulation is or comprises a tablet formulation, and / or an administered formulation is or comprises a candy formulation, and / or an administered formulation is or comprises a capsule formulation. In many embodiments (e.g., of the foregoing embodiments), an administered composition includes a Vitamin E agent (e.g., as a pharmaceutically acceptable carrier); in various such embodiments, such composition is coated with such Vitamin E agent..

[0037] In some embodiments, the present disclosure provides methods of producing a semaglutide polypeptide formulation for oral delivery, the method comprising a step of mixing (1) a semaglutide polypeptide composition (e.g., a crystalline semaglutide polypeptide composition which may, for example, be in powder form); and (2) a pharmaceutically acceptable carrier. In some embodiments, a semaglutide formulation produced by a provided method is or comprises a tablet formulation. In some embodiments, a semaglutide formulation produced by a provided method is or comprises a globule formulation. In some embodiments, a semaglutide formulation produced by a provided method is or comprises a capsule formulation In some embodiments, a semaglutide formulation produced by a provided method is or comprises a truffle formulation. In some embodiments, a semaglutide formulation produced by a provided method is or comprises a candy formulation.

[0038] In some embodiments, the present disclosure provides methods of producing a truffle formulation for oral delivery, including, for example methods comprising a step of placing within a pharmaceutically acceptable shell a core comprising (a crystalline semaglutide polypeptide composition (e.g., a powder form that includes a crystallized polypeptide).

[0039] In some embodiments, the present disclosure provides methods of producing a candy formulation for oral delivery, the method comprising a step of mixing (1) a semaglutide polypeptide composition (e.g., a crystalline semaglutide polypeptide composition which may, for example, be in powder form); and (2) a pharmaceutically acceptable carrier, optionally wherein a pharmaceutically acceptable carrier comprises food ingredients, e.g., chocolate, cocoa, milk, milk product, natural dye, artificial dye, gum base, flavors, sweeteners, gelatin, starch, syrup, citric acid, sugar, etc.

[0040] In some embodiments, provided composition includes a carrier polypeptide composition and in some such embodiments, a carrier polypeptide composition and a semaglutide polypeptide composition of a provided formulation (e.g., truffle, globule, tablet, candy, capsule, etc.) are produced together in a crystallization process. In some embodiments, a carrier polypeptide composition and a semaglutide polypeptide composition of a provided formulation (e.g., truffle, globule, tablet, candy, capsule, etc.) are produced together during a process of crystallizing a carrier polypeptide composition, wherein a semaglutide polypeptide composition is embedded in a carrier polypeptide composition in such process.

[0041] In certain embodiments, a semaglutide polypeptide composition amenable to formulation as described herein may be or comprise a therapeutic semaglutide polypeptide. In some embodiments, a semaglutide polypeptide composition amenable to formulation as described herein may include one or more covalent modifications such as, for example, one or more of acetylation, acylation, amidation, fatty acids, glycation, glycosylation, lipidation, mannosylation, methylation, nitrosylation, phosphorylation, sulfation, palmitoylation, pegylation, prenylation, or a combination thereof.

[0042] In certain embodiments, a carrier polypeptide composition amenable to formulation as described herein may be or comprise a therapeutic semaglutide polypeptide. In some embodiments, a carrier polypeptide composition amenable to formulation as described herein may include one or more covalent modifications such as, for example, one or more of acetylation, acylation, amidation, fatty acids, glycation, glycosylation, lipidation, mannosylation, methylation, nitrosylation, phosphorylation, sulfation, palmitoylation, pegylation, prenylation, or a combination thereof.

[0043] In certain embodiments, a carrier polypeptide composition amenable to formulation as described herein may be or comprise a non-therapeutic polypeptide. In certain embodiments, a carrier polypeptide composition amenable to formulation as described herein may be or comprise a non-therapeutic polypeptide which is otherwise beneficial to therapeutic outcome when utilized in combination of a provided active semaglutide polypeptide composition.

[0044] In some embodiments, a polypeptide included in a carrier polypeptide composition is or comprises, for example, selected from albumins, gluten, globulins, glutenin, prolamin, legumin, vicillin, glycinin, conglycinin, gliadins, myoglobulins, collagen, elastin, myosin, actin, tropomysin, troponin, alpha, beta and kappa caseins, alpha and beta zein, convicillin, gelatin, ovalbumin, sericin, fibroin, beta lactoglobulin, glutelins, helianthinin, immunoglobulins, ovomucoid, ovomucin lysozyme, or ovotransferrin, or a combination thereof.

[0045] In some embodiments, a carrier polypeptide is or comprises amylase. In some embodiments, a carrier polypeptide is or comprises albumin.

[0046] Among other contributions, the present disclosure provides an insight that provided formulations include a polypeptide component (e.g., a carrier polypeptide composition, a semaglutide polypeptide composition) that, in some embodiments, is a singlepolypeptide that is in a crystallized form. Alternatively, in some embodiments, a polypeptide in a provided formulation (e.g., a carrier polypeptide, a semgalutide polypeptide ) is a combination of two or more polypeptides; in some such embodiments, polypeptides are in a crystalline or amorphous form but in some embodiments, not all polypeptides in such polypeptide component need to be crystalline. That is, in some embodiments, less than all of the polypeptides in a polypeptide component of a provided formulation are crystalline. Without wishing to be bound by any particular theory, the present disclosure proposes that one (or more than one) polypeptide in a polypeptide component can act as a crystallization carrier with respect to one (or more than one) other polypeptide in the component. In many provided embodiments, however, a semaglutide polypeptide is crystalline.

[0047] In certain embodiments, a polypeptide component of a provided formulation is or comprises a semaglutide polypeptide, or derivative variant thereof, optionally where the polypeptide is natural, synthetic, or engineered.

[0048] In certain embodiments, a carrier polypeptide amenable to formulation as described herein (e.g., a carrier polypeptide or a semaglutide polypeptide) has a molecular weight between about 100 Da and about 25 kDa, optionally where the molecular weight is between about 100 Da and about 1 kDa, about 100 Da and about 2 kDa, about 100 Da and about 3 kDa, about 100 Da and about 4 kDa, about 100 Da and about 5 kDa, about 100 Da and about 10 kDa, about 100 Da and about 15 kDa, or about 100 Da and about 20 kDa. In certain embodiments, a polypeptide has a molecular weight between about 25 kDa and about 1,000 kDa, optionally where the molecular weight is between about 25 kDa and about 500 kDa, about 100 kDa and about 500 kDa, about 120 kDa and about 250 kDa, or about 150 kDa and about 300 kDa.

[0049] Wild Type semaglutide has a molecular weight of about 4113 Da and in some embodiments, a semagltuide polypeptide used in accordance with the present disclosure may, for example, have a molecular weight between about 3 kDa and about 6 kDA.

[0050] In certain embodiments, a polypeptide formulation provided herein includes about 1 pg to about 2,000 mg of carrier polypeptide (e.g., of a carrier polypeptide composition, which may, in some embodiments, be a single polypeptideand, and when a total formulation is about 5 mg to about 200 g) or active semaglutide polypeptide (e.g., of a polypeptide composition, which may, in some embodiments, be a single polypeptide, and when a total formulation is about 5 mg to about 200 g), optionally where the polypeptideformulation includes about 1 pg to about 1,000 mg, about 1 pg to about 500 mg, about 1 pg to about 400 mg, about 1 pg to about 300 mg, about 1 pg to about 200 mg, about 1 pg to about 100 mg, about 1 pg to about 50 mg, about 1 pg to about 25 mg, about 1 pg to about 20 mg, about 1 pg to about 15 mg, about 1 pg to about 10 mg, about 1 pg to about 5 mg, about 1 gig to about 1 mg, about 1 pg to about 500 pg, about 1 pg to about 250 pg, about 1 pg to about 200 |ag, about 1 pg to about 150 pg, about 1 pg to about 100 pg, about 1 pg to about 50 |jg, about 1 mg to about 1,000 mg, about 1 mg to about 500 mg, about 1 mg to about 400 mg, about 1 mg to about 300 mg, about 1 mg to about 200 mg, about 1 mg to about 100 mg, about 1 mg, to about 50 mg, about 1 mg to about 25 mg of carrier polypeptide (e.g., of a carrier polypeptide composition, which may, in some embodiments, be a single polypeptide) or active semaglutide polypeptide (e.g., of a semaglutide polypeptide composition, which may, in some embodiments, be a single polypeptide).

[0051] In certain embodiments, a provided formulation includes crystals of one or more polypeptides (e.g., those in a carrier polypeptide composition, those in a semaglutide polypeptide composition) having an average particle size of less than 25 microns, e.g., less than 20, 15, 10, 5, 4, 3, 2, 1, or 0.5 microns, optionally where the crystals of semaglutide polypeptide have an average particle size that is between 0.5 microns and 1, 2, 3, 4, 5, 10, 15, 20, or 25 microns or where the crystals of semaglutide polypeptide have an average particle size that is between 1 micron and 1, 2, 3, 4, 5, 10, 15, 20, or 25 microns.

[0052] In certain embodiments, a provided formulation includes crystallized semaglutide polypeptide particles, e.g., having an average particle size of less than 25 microns, e.g., less than 20, 15, 10, 5, 4, 3, 2, 1, or 0.5 microns, optionally where the particles have an average particle size that is between 0.5 microns and 1, 2, 3, 4, 5, 10, 15, 20, or 25 microns or where such particles have an average particle size that is between 1 micron and 1, 2, 3, 4, 5, 10, 15, 20, or 25 microns.

[0053] In certain embodiments, a carrier polypeptide composition of a provided polypeptide formulation includes lyophilized polypeptide. In certain embodiments, a carrier polypeptide composition of a provided polypeptide formulation includes microcrystals of polypeptide. In certain embodiments, a carrier polypeptide composition of a provided polypeptide formulation includes a powder including crystallized polypeptide.

[0054] In certain embodiments, a semaglutide polypeptide composition of a provided polypeptide formulation includes lyophilized semaglutide polypeptide. In certainembodiments, a semaglutide polypeptide composition of a provided polypeptide formulation includes microcrystals of semaglutide polypeptide. In certain embodiments, a semaglutide polypeptide composition of a provided polypeptide formulation includes a powder including crystallized semaglutide polypeptide.

[0055] In certain embodiments, a core (e.g., of a truffle, tablet, or capsule formulation) is a viscous solution. In certain embodiments, such a core comprises crystallized carrier polypeptide composition. In certain embodiments, such a core comprises crystallized semaglutide polypeptide composition. In certain embodiments, a core comprises amorphous carrier polypeptide composition.

[0056] In certain embodiments, a shell (e.g., of a truffle, table, or a capsule formulation) is or comprises sugar. In certain embodiments, a shell is or comprises cane sugar. In certain embodiments, a shell is or comprises palm sugar. In certain embodiments, a shell is or comprises lactose. In certain embodiments, a shell is or comprises xylitol. In certain embodiments, a shell is or comprises milk sugar. In certain embodiments, a shell is cane sugar. In certain embodiments, a shell is palm sugar. In certain embodiments, a shell is hollow. In certain embodiments, a shell is hollow and a core is situated in empty space of a shell. In certain embodiments, a truffle formulation comprises a shell and a core, wherein a shell is hollow and a core is situated in empty space of a shell. In certain embodiments, a truffle shell is hollow and shell is fully filled with space of a core. In certain embodiments, a truffle shell is hollow and the shell is partially filled with space of a core.

[0057] In certain embodiments, a semaglutide polypeptide formulation is formulated for delivery via the gut, optionally where a semaglutide polypeptide formulation is a tablet. In certain embodiments, a semaglutide polypeptide formulation is formulated for delivery via the stomach and / or intestine. In certain embodiments, a semaglutide polypeptide formulation includes an enteric coating. In certain embodiments, a semaglutide polypeptide formulation includes a Vitamin E coating.

[0058] In certain embodiments, a provided polypeptide formulation, in addition to a semaglutide polypeptide composition; and optionally a pharmaceutically acceptable carrier (and further optionally a carrier polypeptide composition) comprises one or more excipients or additives selected from the group consisting of aggregation-reducing agents, sugars or sugar alcohols, polysaccharides, stabilizers, hyaluronidase, buffering agents, preservatives, carriers, antioxidants, chelating agents, natural or synthetic polymers, cryoprotectants,lyoprotectants, surfactants, bulking agents, acidifying agents, ingredients to reduce injection site discomfort, antifoaming agents, alkalizing agents, vehicles, aggregation inhibitors, solubilizing agents, tonicity modifiers, permeation enhancers, muco bioadhesive agents, and stabilizing agents and combinations thereof.

[0059] In certain embodiments, one or more excipients or additives are individually or cumulatively present in a provided formulation at a concentration between 0.1 mM and about 1,000 mM, between about 0.1 mM and about 500 mM, between about 0.1 mM and about 200 mM, or between about 0.1 mM and about 100 mM.

[0060] In certain embodiments, a provided semaglutide formulation includes one or more aggregation-reducing agent(s)s, such as may be selected from the group consisting of nicotinic acid, caffeine citrate, caffeine nicotinate, caffeine, octyl- P-D-glucopyranoside, and n-dodecyl-P-D-maltoside and optionally in combination with one or more of arginine, tryptophan, histidine, proline, cysteine, methionine, P-alanine, Potassium Glutamate, Arginine Ethylester, lysine, aspartic acid, glutamic acid, glycine, DTPA (diethylenetriaminepentaacetic acid), EGTA(aminopolycarboxylic acid), EDTA (Ethylenediaminetetraacetic acid), hydroxy propyl beta (HP-Beta) cyclodextrins, hydroxy propyl gamma (HP-Gamma) cyclodextrins, sulfo-butyl ether (SBE) cyclodextrins, TMAO (trimethylamine N-oxide), trehalose, ethylene glycol, betaine, xylitol, sorbitol, 6-(N-(7- nitrobenz-2-oxa-l,3-diazol-4-yl)amino)hexanoic acid (NBD-X), methyl acetyl phosphate (MAP), citraconic anhydride, pyrophosphate, citrate, and combinations thereof.

[0061] Alternatively or additionally, in some embodiments, a provided semaglutide formulation may include a tonicity modifier(s), for example which may be selected from the group consisting of arginine, cysteine, histidine, glycine, sodium chloride, potassium chloride, sodium citrate, saccharides such as sucrose, glucose, dextrose, glycerin or mannitol, and combinations thereof.

[0062] Alternatively or additionally, in some embodiments, a provided semaglutide formulation may include an antioxidant(s), for example which may be selected from the group consisting of glycine, lysine, EDTA, DTPA, sorbitol, mannitol, ascorbic acid, ascorbyl palmitate, butylated hydroxyanisole, butylated hydroxytoluene, hypopho sphorous acid, monothioglycerol, propyl gallate, sodium bisulfite, sodium formaldehyde sulfoxylate, sodium metabisulfite, sodium thiosulfate, sufur dioxide, tocopherol, and combinations thereof.

[0063] Alternatively or additionally, in some embodiments, a provided semaglutide formulation may include a lyoprotectant(s), for example which may be selected from the group consisting of sucrose, lactose, trehalose, dextran, erythritol, arabitol, xylitol, sorbitol, maltose, lactulose, maltulose, glucitol, maltitol, lactitol, isomaltulose and mannitol; amino acids, such as arginine or histidine or proline or glycine; lyotropic salts, such as magnesium sulfate; propylene glycol, glycerol, poly(ethylene glycol), or polypropylene glycol); gelatin, dextrins, modified starch, carboxymethyl cellulose, and combinations thereof.

[0064] Alternatively or additionally, in some embodiments, a provided semaglutide formulation may include a permeation enhancer(s), for example which may be selected from the group consisting of bile salts, e.g., tri-hydroxy salts sodium cholate, sodium glycocholate, sodium taurocholate and di-hydroxy salt, sodium deoxy cholate, sodium glyco-deoxy Cholate, sodium tauro-deoxy cholate; fatty acids, their salt and esters, e.g., oleic acid, lauric acid, cod liver oil extract, sodium laurate, sodium caprate, glyceryl monostearate, di-ethylene glycol mono ethyl ether and various sucrose fatty acid esters, medium-chain fatty acid glycerides, polycaprolactoneomega-3 fatty acids, lecithin (phosphatidylcholine), lysophosphatidylcholine; surfactants, e.g., sodium dodecyl (lauryl) sulphate, poly sorbates (polysorbate 80), laureths, brijs and benzalkonium chloride; complexing agents, e.g., cyclodextrins, dextran sulphate, dextran sulphate, sodium edetate; complexing agents, e.g., cyclodextrins, dextran sulphate, dextran sulphate, sodium edetate, co-solvents, e.g., ethanol and propylene glycol, combination of 1% oleic acid and 5% / 10% polyethylene glycol 200, 2% glyceryl mono laurate and 40% alcohol, sodium caprate and alcohol or propylene glycol, 10% lauric acid in propylene glycol, polyoxyethylene, 2,3-lauryl ether, menthol, sodium caprate, sodium caprylate, sodium glycodeoxycholate, glycol; polysaccharides, e.g., chitosan and chitosan glutamate; and others such as, aprotinin, benzalkonium chloride, cetylpyridinium chloride, cetyltrimethyl ammonium bromide, sodium salicylate, lysophosphatidylcholine, methoxysalicylate, methyloleate , sodium edta, sulfoxides, various alkyl glycosides, ethylene-diamide tetra acetic acid (edta), tartaric acid; lyotropic salts, such as magnesium sulfate; propylene glycol, glycerol, poly(ethylene glycol), or polypropylene glycol); gelatin, dextrins, modified starch, carboxymethyl cellulose, and combinations thereof.

[0065] Alternatively or additionally, in some embodiments, a provided semaglutide formulation may include an absorption enhancer(s), for example which may be selected from the group consisting of surfactants, cholesterol, glycerides, salicylates, bile salts, chelatingagents, sodium caprate, a salt of capric acid and other includes N-(5-chlorosalicylol)-8- aminocaprylic acid (5-CNAC), 4-((4-chloro-2-hydroxybenzoyl))-amino) butanoic acid (4- CNAB) and N-(8-(2-hydroxybenzoyl))-amino) caprylic acid, also known as salcaprozate sodium (SNAC, caprylic acid, C8, castor oil, medium chain, acyl carnitine, EDTA, glyceryl monolaurate, bovine P-casein, tocopherol succinate glycol chitosan conjugates, lecithins, glyceryl monostearate (GMS), chitosan and alginate, PLGA, silica, stearic acid, oleic acid, hydrogenated castor oil, and glyceryl trimyristate, etoposide phosphate (Vepesid®), sulindac (Clinoril®), enalapril maleate (Vasotec®), ramipril (Altace®), olmesartan medoxomil (Benicar®), valacyclovir (Valtrex®), midodrine (Amatine®), gabapentin enacarbil (Horizant®), or sulfasalazine (Azulfidine®).

[0066] Alternatively or additionally, in some embodiments, a provided semaglutide formulation may include a muco bioadhesive agent(s), for example which may be selected selected from the group consisting of sucrose, lactose, trehalose, dextran, erythritol, arabitol, xylitol, sorbitol, maltose, lactulose, maltulose, glucitol, maltitol, lactitol, isomaltulose and mannitol; amino acids, such as arginine or histidine or proline or glycine; lyotropic salts, such as magnesium sulfate; propylene glycol, glycerol, poly(ethylene glycol), or polypropylene glycol); gelatin, dextrins, modified starch, carboxymethyl cellulose, and combinations thereof, mucoadhesive system such as from naturae, e.g., gelatin, agarose, chitosan, hyaluronic acid and synthetic polymers, e.g., polyvinylpyrolidone (PVP), polycrylates, polyvinyl alcohol, sodium carboxymethyl cellulose (SCMC) and pectin, all anionic-type polymers, chitosan (cationic type), and hydroxypropyl methylcellulose (HPMC) as a nonionic polymer, polyacrylic acid (PAA) derivatives (CP934, CP940, PCP), 15% CMC and 35% CP, copolymers of acrylic acid and poly(ethylene glycol) monomethylether monomethacrylate (PEGMM), eudragitl NE40D is a neutral poly(ethylacrlate methylmethacrylate, hydrophilic polymers, e.g., methocel K4M, methocel K15M, SCMC 400, Cekol 700, Cekol 10000, CP934P, CP971P and CP974P, carboxyvinyl polymer and triethanolamine, HPC(hydroxy propyl celluose), CP (carbopol 934P), carbopol (CP) EX-55 CMC (sodium carboxymethyl cellulose), HPMC (hydroxy propyl methyl cellulose), HEC (hydroxy ethyl cellulose), PIP [poly(isoprene)], PIB [poly(Isobutylene)], xanthum gum, locust bean gum, pectin, polycarbophil, benzyl esters, hydroxyethylcellulose, poly(acrylic acid), poly(acrylic acid-co- acrylamide), poly(acrylic acid-co-methyl methacrylate), poly(acrylic acid-co-butylacrylate), HEMA copolymerized with Polymeg® (polytetramethylene glycol), Cydot® (bioadhesive polymeric blend of CP and PIB), formulation consisting of PVP, cetylpyridinium chloride (asstabilizer), chitosan chloride, polyethylene oxide, polymethylvinylether / maleic anhydride (PME / MA), and tragacanth, poly ethyleneglycol monomethylether monomethacrylate, drum dried waxy maize starch (DDWM), carbopol 974P, and sodium stearylfumarate, and cellulose derivatives; hyderogels-acrylic acid (polar) and butyl acrylate (apolar), and combinations thereof.DEFINITIONS

[0067] A, An, The: As used herein, “a”, “an”, and “the” refer to one or to more than one (z.e., to at least one) of the grammatical object of the article. By way of example, “an element” discloses embodiments of exactly one element and embodiments including more than one element.

[0068] About: As used herein, term “about”, when used in reference to a value, refers to a value that is similar, in context to the referenced value. In general, those skilled in the art, familiar with the context, will appreciate the relevant degree of variance encompassed by “about” in that context. For example, in some embodiments, the term “about” may encompass a range of values that within 25%, 20%, 19%, 18%, 17%, 16%, 15%, 14%, 13%, 12%, 11%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, or less of the referenced value.

[0069] Administration: As used herein, the term “administration” typically refers to administration of a composition to a subject or system to achieve delivery of an agent that is, or is included in, the composition.

[0070] Agent. As used herein, the term “agent” may refer to any chemical entity, including without limitation any of one or more of an atom, molecule, compound, amino acid, polypeptide, nucleotide, nucleic acid, polypeptide complex, liquid, solution, saccharide, polysaccharide, lipid, or combination or complex thereof.

[0071] Amino acid: In its broadest sense, as used herein, refers to any compound and / or substance that can be incorporated into a polypeptide chain, e.g., through formation of one or more peptide bonds. In some embodiments, an amino acid has the general structure H2N-C(H)(R)-COOH. In some embodiments, an amino acid is a naturally -occurring amino acid. In some embodiments, an amino acid is a non-natural amino acid; in some embodiments, an amino acid is a D-amino acid; in some embodiments, an amino acid is an L- amino acid. “Standard amino acid” refers to any of the twenty standard L-amino acids commonly found in naturally occurring peptides. “Nonstandard amino acid” refers to any amino acid, other than the standard amino acids, regardless of whether it is preparedsynthetically or obtained from a natural source. In some embodiments, an amino acid, including a carboxy- and / or amino-terminal amino acid in a polypeptide, can contain a structural modification as compared with a typical or canonical amino acid structure. For example, in some embodiments, an amino acid can be modified by methylation, amidation, acetylation, pegylation, glycosylation, phosphorylation, and / or substitution (e.g., of the amino group, the carboxylic acid group, one or more protons, and / or the hydroxyl group) as compared with the general structure. In some embodiments, such modification can, for example, alter the circulating half-life of a polypeptide containing the modified amino acid as compared with one containing an otherwise identical unmodified amino acid. In some embodiments, such modification does not significantly alter a relevant activity of a polypeptide containing the modified amino acid, as compared with one containing an otherwise identical unmodified amino acid. As will be clear from context, in some embodiments, the term “amino acid” can be used to refer to a free amino acid; in some embodiments it can be used to refer to an amino acid residue of a polypeptide.

[0072] Amorphous’. As used herein, the term “amorphous” generally refers to a noncrystalline solid form of polypeptide, sometimes referred to as an amorphous solid” or “amorphous precipitate”, which typically has no, or essentially no, molecular lattice structure characteristic of the crystalline solid state.

[0073] Analog: As used herein, the term “analog” refers to a substance that shares one or more particular structural features, elements, components, or moieties with a reference substance. Typically, an “analog” shows significant structural similarity with the reference substance, for example sharing a core or consensus structure, but also differs in certain discrete ways. In some embodiments, an analog is a substance that can be generated from the reference substance, e.g., by chemical manipulation of the reference substance. In some embodiments, an analog is a substance that can be generated through performance of a synthetic process substantially similar to (e.g., sharing a plurality of steps with) one that generates the reference substance. In some embodiments, an analog is or can be generated through performance of a synthetic process different from that used to generate the reference substance.

[0074] Associated with: Two events or entities are “associated” with one another, as that term is used herein, if the presence, level and / or form of one is correlated with that of the other. For example, a particular entity (e.g., polypeptide, genetic signature, metabolite, microbe, etc.) is considered to be associated with a particular disease, disorder, or condition, if its presence, level and / or form correlates with incidence of and / or susceptibility to thedisease, disorder, or condition (e.g., across a relevant population). In some embodiments, two or more entities are physically “associated” with one another if they interact, directly or indirectly, so that they are and / or remain in physical proximity with one another. In some embodiments, two or more entities that are physically associated with one another are covalently linked to one another; in some embodiments, two or more entities that are physically associated with one another are not covalently linked to one another but are non- covalently associated, for example by means of hydrogen bonds, van der Waals interaction, hydrophobic interactions, magnetism, or a combination thereof.

[0075] Between or From’. As used herein, the term “between” refers to content that falls between indicated upper and lower, or first and second, boundaries (or “bounds”), inclusive of the boundaries. Similarly, the term “from”, when used in the context of a range of values, indicates that the range includes content that falls between indicated upper and lower, or first and second, boundaries, inclusive of the boundaries.

[0076] Bioavailability’. As used herein, the term “bioavailability” can refer to the degree to which a substance, e.g., a polypeptide such as smeaglutide or a semaglutide fragment, administered to an in vivo subject, becomes available to a tissue to which the substance is targeted (e.g., the bloodstream and / or plasma). Bioavailability can refer to the degree to which a substance that has been administered to an in vivo subject is delivered to blood of the subject. Bioavailability can refer to the ability of a substance to perform a function in the subject. Bioavailability can be measured in a number of ways, e.g., as the concentration of a substance in the bloodstream or plasma. In some embodiments, bioavailability can be assessed, for example, by comparing the “area under the curve” (AUC) in a plot of the plasma concentration as a function of time (area under the plasma concentration curve from time zero to a time where the plasma concentration returns to baseline levels). AUC can be calculated, for example, using the linear trapezoidal rule.“AUCO-f ’ refers to the area under the plasma concentration curve from time zero to a time, t, later, for example to the time of reaching baseline.

[0077] Cancer: As used herein, the term “cancer” refers to a disease, disorder, or condition in which cells exhibit relatively abnormal, uncontrolled, and / or autonomous growth, so that they display an abnormally elevated proliferation rate and / or aberrant growth phenotype characterized by a significant loss of control of cell proliferation. In some embodiments, a cancer can include one or more tumors. In some embodiments, a cancer can be or include cells that are precancerous (e.g., benign), malignant, pre-metastatic, metastatic,and / or non-metastatic. In some embodiments, a cancer can be or include a solid tumor. In some embodiments, a cancer can be or include a hematologic tumor.

[0078] Carrier polypeptide: As used herein, the term “carrier polypeptide” refers to a polypeptide that is crystallized or otherwise crystalline as used in the present disclosure. In some embodiments, a carrier polypeptide may have biological activity (e.g., may be considered an “active” in a drug product). In some embodiments, a carrier polypeptide may not have biological activity (e.g., may not have biological activity sufficient to be considered an “active” in a drug product”). In some particular embodiments, a biologically inactive carrier polypeptide may be selected from the group consisting of actin, albumins, caseins (e.g., alpha, beta and kappa caseins), collagen, conglycinin, convicillin, elastin, fibroin, gelatin, gliadins, globulins, glutelins, gluten, glutenin, glycinin, helianthinin, immunoglobulins, lactoglobulin (e.g., beta lactoglobulin), legumin, lysozyme, myoglobulins, myosin, ovalbumin, ovomucin, ovomucoid, ovotransferrin, prolamin, sericin, tropomysin, troponin, vicillin, zein (e.g., alpha and beta zein).

[0079] Crystalline’. As used herein, the term “crystalline” or “crystal” refers to a material whose constituents (such as atoms, molecules, or ions) are arranged in a highly ordered microscopic structure, forming a crystal lattice that extends in all directions. In many embodiments, presence of crystalline material can be detected, for example, by X-ray diffraction (e.g., by X-ray Powder Diffraction analysis, XRPD).

[0080] Engineered’. As used herein, the term “engineered” refers to the aspect of having been manipulated by the hand of man. For example, a polynucleotide is considered to be “engineered” when two or more sequences, that are not linked together in that order in nature, are manipulated by the hand of man to be linked to one another in the engineered polynucleotide. Those of skill in the art will appreciate that an “engineered” nucleic acid or amino acid sequence can be a recombinant nucleic acid or amino acid sequence. In some embodiments, an engineered polynucleotide includes a coding sequence and / or a regulatory sequence that is found in nature operably linked with a first sequence but is not found in nature operably linked with a second sequence, which is in the engineered polynucleotide and operably linked in with the second sequence by the hand of man. In some embodiments, a cell or organism is considered to be “engineered” if it has been manipulated so that its genetic information is altered (e.g., new genetic material not previously present has been introduced, for example by transformation, mating, somatic hybridization, transfection, transduction, or other mechanism, or previously present genetic material is altered or removed, for example by substitution, deletion, or mating). As is common practice and is understood by those ofskill in the art, progeny or copies, perfect or imperfect, of an engineered polynucleotide or cell are typically still referred to as “engineered” even though the direct manipulation was of a prior entity.

[0081] Excipient: As used herein, “excipient” refers to a non-therapeutic agent that may be included in a pharmaceutical composition, for example to provide or contribute to a desired consistency or stabilizing effect. In some embodiments, suitable pharmaceutical excipients may include, for example, starch, glucose, lactose, sucrose, gelatin, malt, rice, flour, chalk, silica gel, sodium stearate, glycerol monostearate, talc, sodium chloride, dried skim milk, glycerol, propylene, glycol, water, ethanol, or the like.

[0082] "Improve " "increase " "inhibit " or “reduce”' As used herein, the terms “improve”, “increase”, “inhibit”, and “reduce”, and grammatical equivalents thereof, indicate qualitative or quantitative difference from a reference.

[0083] Muco bioadhesive agent: As used herein, the term “muco bioadhesive agent” refers to an agent which increases the contact time between a material (e.g., a drug, an active ingredient of a drug, a polypeptide formulation as described herein or polypeptide released therefrom) and mucus or mucous membrane. In some embodiments, a muco bioadhesive agent prolongs the retention of a material at a site of application. In some embodiments, a muco bioadhesive agent alters the rate of release of a material for improved therapeutic outcome.

[0084] Oral administration: As used herein, the term “oral administration” refers to a route of administration where a substance is taken through the mouth. In some embodiments, oral administration is or comprises buccal administration. In some embodiments, oral administration is or comprises sublingual administration. In some embodiments, oral administration is or comprises spray to mouth. In some embodiments, oral administration is via a patch. In some embodiments, oral administration is via application of a film. In some embodiments, oral administration involves administration of drops. In some embodiments, oral administration is or comprises application of a gels. In some embodiments, oral administration is via a wafer. In some embodiments, oral administration is via a capsule. In some embodiments, oral administration is via a tablet. In some embodiments, oral administration is via a suspension. In some embodiments, oral administration is via a formulation as described herein.

[0085] Permeation enhancer: As used herein, the term “permeation enhancer” refers to an agent whose presence or level correlates with improved transport of a material e.g., a drug product or active ingredient thereof, a polypeptide formulation as described herein or apolypeptide component thereof, etc, across an epithelial barrier. In some embodiments, an epithelial barrier is or comprises a mucosal membrane.

[0086] Pharmaceutically acceptable: As used herein, the term “pharmaceutically acceptable,” as applied to one or more, or all, component(s) for formulation of a composition as disclosed herein, means that each component must be compatible with the other ingredients of the composition and not deleterious to the recipient thereof.

[0087] Pharmaceutical composition or formulation’. As used herein, the term“pharmaceutical composition” or “formulation” refers to a composition in which a therapeutic agent is formulated together with one or more pharmaceutically acceptable carriers.

[0088] Polypeptide: As used herein, “polypeptide” refers to any polymeric chain of two or more amino acids. In some embodiments, a polypeptide has an amino acid sequence that occurs in nature. In some embodiments, a polypeptide has an amino acid sequence that does not occur in nature. In some embodiments, a polypeptide has an amino acid sequence that is engineered in that it is designed and / or produced through action of the hand of man. In some embodiments, a polypeptide may be or include of natural amino acids, non-natural amino acids, or both. In some embodiments, a polypeptide may be or include only natural amino acids or only non-natural amino acids. In some embodiments, a polypeptide can include D-amino acids, L-amino acids, or both. In some embodiments, a polypeptide may include only L-amino acids. In some embodiments, a polypeptide may include one or more pendant groups or other modifications, e.g., one or more amino acid side chains, e.g., at the polypeptide’s N-terminus, at the polypeptide’s C-terminus, at non-terminal amino acids, or at any combination thereof. In some embodiments, such pendant groups or modifications may be selected from acetylation, amidation, lipidation, methylation, phosphorylation, glycosylation, glycation, sulfation, mannosylation, nitrosylation, acylation, palmitoylation, prenylation, pegylation, etc., including combinations thereof. In some embodiments, a polypeptide may be cyclic, and / or may include a cyclic portion.

[0089] In some embodiments, the term “polypeptide” may be appended to a name of a reference polypeptide, activity, or structure to indicate a class of polypeptides that share a relevant activity or structure. For such classes, the present specification provides and / or those skilled in the art will be aware of exemplary polypeptides within the class whose amino acid sequences and / or functions are known. In some embodiments, a member of a polypeptide class or family shows significant sequence homology or identity with, shares a common sequence motif (e.g., a characteristic sequence element) with, and / or shares acommon activity (in some embodiments at a comparable level or within a designated range) with a reference polypeptide of the class. For example, in some embodiments, a member polypeptide shows an overall degree of sequence homology or identity with a reference polypeptide that is at least about 30-40%, and is often greater than about 50%, 60%, 70%, 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more and / or includes at least one region (e.g., a conserved region that can in some embodiments be or include a characteristic sequence element) that shows very high sequence identity, often greater than 90% or even 95%, 96%, 97%, 98%, or 99%. Such a conserved region usually encompasses at least 3-4 and in some instances up to 20 or more amino acids; in some embodiments, a conserved region encompasses at least one stretch of at least 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15 or more contiguous amino acids. In some embodiments, a relevant polypeptide can be or include a fragment of a parent polypeptide. In some embodiments, a useful polypeptide may be or include a plurality of fragments, each of which is found in the same parent polypeptide in a different spatial arrangement relative to one another than is found in the polypeptide of interest (e.g., fragments that are directly linked in the parent may be spatially separated in the polypeptide of interest or vice versa, and / or fragments may be present in a different order in the polypeptide of interest than in the parent), so that the polypeptide of interest is a derivative of its parent polypeptide.

[0090] Reference: As used herein, “reference” refers to a standard or control relative to which a comparison is performed. For example, in some embodiments, an agent, sample, sequence, subject, animal, or individual, or population thereof, or a measure or characteristic representative thereof, is compared with a reference, an agent, sample, sequence, subject, animal, or individual, or population thereof, or a measure or characteristic representative thereof. In some embodiments, a reference is a measured value. In some embodiments, a reference is an established standard or expected value. In some embodiments, a reference is a historical reference. A reference can be quantitative of qualitative. Typically, as would be understood by those of skill in the art, a reference and the value to which it is compared represents measure under comparable conditions. Those of skill in the art will appreciate when sufficient similarities are present to justify reliance on and / or comparison. In some embodiments, an appropriate reference may be an agent, sample, sequence, subject, animal, or individual, or population thereof, under conditions those of skill in the art will recognize as comparable, e.g., for the purpose of assessing one or more particular variables (e.g., presence or absence of an agent or condition), or a measure or characteristic representative thereof.

[0091] Small molecule: As used herein, the term “small molecule” means a low molecular weight organic and / or inorganic compound. In general, a “small molecule” is a molecule that is less than about 5 kilodaltons (kD) in size. In some embodiments, a small molecule is less than about 4 kD, 3 kD, about 2 kD, or about 1 kD. In some embodiments, the small molecule is less than about 800 daltons (D), about 600 D, about 500 D, about 400 D, about 300 D, about 200 D, or about 100 D. In some embodiments, a small molecule is less than about 2,000 g / mol, less than about 1500 g / mol, less than about 1,000 g / mol, less than about 800 g / mol, or less than about 500 g / mol. In some embodiments, a small molecule is not a polymer. In some embodiments, a small molecule does not include a polymeric moiety. In some embodiments, a small molecule is not and / or does not include a polypeptide. In some embodiments, a small molecule is not and / or does not include a polynucleotide (e.g., is not an oligonucleotide). In some embodiments, a small molecule is not and / or does not include a polysaccharide; for example, in some embodiments, a small molecule is not a glycoprotein, proteoglycan, glycolipid, etc.). In some embodiments, a small molecule is not a lipid. In some embodiments, a small molecule is a modulating agent (e.g., is an inhibiting agent or an activating agent). In some embodiments, a small molecule is biologically active. In some embodiments, a small molecule is detectable (e.g., includes at least one detectable moiety). In some embodiments, a small molecule is a therapeutic agent.

[0092] Subject: As used herein, the term “subject” refers to an organism, typically a mammal (e.g., a human, rat, or mouse). In some embodiments, a subject is suffering from a disease, disorder or condition. In some embodiments, a subject is susceptible to a disease, disorder, or condition. In some embodiments, a subject displays one or more symptoms or characteristics of a disease, disorder or condition. In some embodiments, a subject is not suffering from a disease, disorder or condition. In some embodiments, a subject does not display any symptom or characteristic of a disease, disorder, or condition. In some embodiments, a subject has one or more features characteristic of susceptibility to or risk of a disease, disorder, or condition. In some embodiments, a subject is a subject that has been tested for a disease, disorder, or condition, and / or to whom therapy has been administered. In some instances, a human subject can be interchangeably referred to as a “patient” or “individual.” A subject administered an agent associated with treatment of a disease, disorder, or condition with which the subject is associated can be referred to as a subject in need of the agent, i.e., as a subject in need thereof.

[0093] Therapeutic agent: As used herein, the term “therapeutic agent” refers to any agent that elicits a desired pharmacological effect when administered to a subject. In someembodiments, an agent is considered to be a therapeutic agent if it demonstrates a statistically significant effect across an appropriate population. In some embodiments, the appropriate population can be a population of model organisms or a human population. In some embodiments, an appropriate population can be defined by various criteria, such as a certain age group, gender, genetic background, preexisting clinical conditions, etc. In some embodiments, a therapeutic agent is a substance that can be used for treatment of a disease, disorder, or condition. In some embodiments, a therapeutic agent is an agent that has been or is required to be approved by a government agency before it can be marketed for administration to humans. In some embodiments, a therapeutic agent is an agent for which a medical prescription is required for administration to humans.

[0094] Therapeutically effective amount: As used herein, “therapeutically effective amount” refers to an amount that produces the desired effect for which it is administered. In some embodiments, the term refers to an amount that is sufficient, when administered to a population suffering from or susceptible to a disease, disorder, and / or condition in accordance with a therapeutic dosing regimen, to treat the disease, disorder, and / or condition. In some embodiments, a therapeutically effective amount is one that reduces the incidence and / or severity of, and / or delays onset of, one or more symptoms of the disease, disorder, and / or condition. Those of ordinary skill in the art will appreciate that a therapeutically effective amount does not necessarily achieve successful treatment in every particular treated individual. Rather, a therapeutically effective amount may be that amount that provides a particular desired pharmacological response in a significant number of subjects when administered to patients in need of such treatment. In some embodiments, reference to a therapeutically effective amount may be a reference to an amount as measured in one or more specific tissues (e.g., a tissue affected by the disease, disorder or condition) or fluids (e.g., blood, saliva, serum, sweat, tears, urine, etc.). Those of ordinary skill in the art will appreciate that, in some embodiments, a therapeutically effective amount of a particular agent or therapy may be formulated and / or administered in a single dose. In some embodiments, a therapeutically effective agent may be formulated and / or administered in a plurality of doses, for example, as part of a dosing regimen.

[0095] Treatment. As used herein, the term “treatment” (also “treat” or “treating”) refers to administration of a therapy that partially or completely alleviates, ameliorates, relieves, inhibits, delays onset of, reduces severity of, and / or reduces incidence of one or more symptoms, features, and / or causes of a particular disease, disorder, or condition, or is administered for the purpose of achieving any such result. In some embodiments, suchtreatment can be of a subject who does not exhibit signs of the relevant disease, disorder, or condition and / or of a subject who exhibits only early signs of the disease, disorder, or condition. Alternatively or additionally, such treatment can be of a subject who exhibits one or more established signs of the relevant disease, disorder and / or condition. In some embodiments, treatment can be of a subject who has been diagnosed as suffering from the relevant disease, disorder, and / or condition. In some embodiments, treatment can be of a subject known to have one or more susceptibility factors that are statistically correlated with increased risk of development of the relevant disease, disorder, or condition.BRIEF DESCRIPTION OF THE DRAWINGFIG. 1 is a set of two images showing hollow sugar truffles.DETAILED DESCRIPTION

[0096] Extensive effort has been expended by the pharmaceutical industry to bring polypeptide therapeutics to market. Clinical trials have included hundreds of polypeptides for treatment of multiple different conditions. However, clinical use of polypeptides has been hampered by numerous obstacles to obtaining suitable forms (e.g., polymorphs of solids) for formulation and their successful delivery. Bioavailability, stability, and therapeutic efficacy are among the factors considered in the development of formulations for polypeptide delivery.

[0097] Semaglutide polypeptides are often administered by parenteral routes, and particularly by subcutaneous injection.

[0098] The present disclosure identifies a source of a problem with prior technologies for administration of semaglutide polypeptides, in their reliance primarily on parenteral strategies. The present disclosure further identifies a source of a problem with conventional strategies (specifically, Rybelsus® tablets) for oral administration of semaglutide, for example relating to inefficient delivery to the bloodstream.

[0099] The present disclosure provides an insight that oral administration is often preferable to injection of semaglutide polypeptide therapeutics, among other things for patient acceptance, home use, and compliance with long-term regimens. Further, the present disclosure provides an insight that use of crystalline semaglutide polypeptide compositions in oral formulations can provide surprising delivery benefits, particularly when a Vitamin E agent is included in the formulation and / or when the formulation is arranged and constructed to rapidly dissolve. Among other things, the present disclosure provides a specific insight that rapidly dissolving formulations, specifically including rapidly dissolving tablets, may be particularly useful for oral administration of semaglutide polyeptides.

[0100] Additionally, the present disclosure encompasses an insight that during a process of crystallizing a carrier polypeptide composition, a semaglutide polypeptide composition can be embedded in such carrier polypeptide composition, thereby creating a composition comprising both a carrier polypeptide composition and a semaglutide polypeptide composition. In some embodiments, a carrier polypeptide and a semaglutide polypeptide form a co-crystal during crystallization. In some embodiments, such composition comprising both a carrier polypeptide composition and a polypeptide composition (e.g., a co-crystal of carrier polypeptide composition and a semaglutide polypeptide composition) is amenable for formulation.

[0101] The present disclosure provides a variety of formulations that achieve systemic delivery of semaglutide polypeptides via oral administration of the formulation. For example, in some embodiments, the present disclosure provides formulations that comprise (1) a semaglutide polypeptide composition (e.g., a crystalline semaglutide polypeptide composition which may, in some embodiments, be a powder); and (2) one or more pharmaceutically acceptable carriers or excipients. In some embodiments, provided oral formulations include (3) a carrier polypeptide composition (e.g., a crystalline carrier polypeptide composition).

[0102] In some embodiments, the present disclosure provides a tablet formulation, optionally wherein a pharmaceutically acceptable carrier of a capsule formulation comprises a Vitamin E agent. In some embodiments, the present disclosure provides a globule formulation. In some embodiments, the present disclosure provides a candy formulation. In some embodiments, the present disclosure provides a capsule formulation, optionally wherein a pharmaceutically acceptable carrier of a capsule formulation comprises a Vitamin E agent.

[0103] The present disclosure provides an insight that polypeptide compositions as described herein, e.g., truffle, globule, tablet, candy, capsule, etc. optionally comprise food ingredients and can be particularly desirable and / or effective for oral administration. In some embodiments, polypeptide compositions comprising food ingredients as described herein have improved flavor, more attractive appearance, and / or are easier to handle compared to other formulations for oral administration, and compared to parental formulations. These advantages among others could cause oral formulations of polypeptide products to have therapeutic and commercial value distinct from and / or greater than those of other formulations, specifically including parenteral formulations.SEMAGLUTIDE

[0104] Semaglutide is a glucagon-like peptide- 1 (GLP-1) receptor agonist that is central to the recent revolution in treatment of diabetes, obesity, and associated conditions.

[0105] Semaglutide has been approved by the United States Food and Drug Administration (FDA) in three different medications: Ozempic®, Rybelsus®, and Wegovy®. Ozempic® is an injectible product, formulated in three different doses (0.5mg, Img, and 2mg) and approved (i) as an adjunct to diet and exercise to improve glycemic control in adults with type 2 diabeted; and (2) to reduce risk of magor adverse cardiovascularevents in adults with type 2 diabetes mellitus and established carfiovascular disease. Rybelsus® is an oral tablet, formulated in threeo different doses (3mg, 7mg and 14mg), also approved as an adjunct to diet and exercise to improve glyceminc control in adults with type 2 diabetes mellitus. Wegovy® is an injectable product, formulated in five different doses (0.25 mg, 0.5 mg, Img, 1.7mg, and 2.4mg), and approved for use in combination with a redsuced calorie diet and increased physical activity (i) to reduce risk of magor adverse cardiovascular events (cardiocascular death, non-fatal myocardial infarction, or non-fatal stroke) in adults with established cardiovascular disease who are either obese or overweight; and (ii) to reduce excels body weight and maintain weight reduction long term in (a) obese subjects 12 years of age and older; and (b) adults who are overweight and have at least one weight-related comorbid condition.

[0106] The present disclosure provides new oral semaglutide polypeptide compositions, and demonstrates surprising characteristics, including relative to Rybelsus®. Among other things, the present disclosure demonstrates that certain compositions that utilize crystalline semaglutide polypeptide rather than amorphous, as is utilized in Rybelsus®, achieve surprising results relative to Rybelsus®. Alternatively or additionally, the present disclosure demonstrates that semaglutide peptide composition that include (e.g., are coated with) Vitamin E provide remarkable attributes, including specifically unexpected delivery attributes.VITAMIN E

[0107] Vitamin E agents, as used herein, refer to compounds or entities that, when administered to a subject, deliver to that subject a Vitamin E active moiety. In some embodiments, a Vitamin E agent is provided and / or utilized as a salt, co-crystal, free acid or base, solvate, ester, hydrate, polymorph, or anhydrous form. In some embodiments, a Vitamin E agent is provided and / or utilized in a particular stereoisomeric form, or as amixture of stereoisomeric forms. In some embodiments, a Vitamin E agent is a prodrug of Vitamin E wherein Vitamin E is the intended metabolite for a therapeutic effect.

[0108] In some embodiments, Vitamin E agents can be or include one or more Vitamin E tocols selected from one or more Vitamin E tocopherols and / or one or more Vitamin E tocotrienols. Natural Vitamin E tocols include two series of compounds: tocopherols with a saturated side chain and tocotrienols with an unsaturated side chain. Tocopherols and tocotrienols have a similar chemical structure, which is characterized by a long isoprenoid side chain attached at the 2 position of a 6-chromanol ring. Tocopherols include a chromanol ring and a 16-carbon tail. Tocotrienols differ from tocopherols in that they possess a farnesyl rather than a saturated isoprenoid C16 side chain. Vitamin E tocols (e.g., tocopherols and tocotrienols) are designated as a, P, y, or 5 based on the methylation pattern of the chromanol ring, p- and y- are dimethylated in the 5- and 8-positions or the 7- and 8-positions of the chromanol ring, respectively, a-tocols are trimethylated in the 5-, 7-, and 8-positions of the chromanol ring, and 5- are monomethylated in the 8-position of the chromanol ring. Vitamin E agents can include an a-, P-, y-, or 5-tocol, or any mixture thereof. For example, Vitamin E tocopherols can include any of one or more of tocopherols a, P, y, and 5, as well as derivatives thereof. Vitamin E tocotrienols can include any of one or more of tocotrienols a, P, y, and 5, as well as derivatives thereof. Of these Vitamin E tocols, a-tocopherol and y-tocopherol are the most abundant in nature. Certain commercially available Vitamin E supplements can commonly include a-tocopherol. Certain commercially available Vitamin E supplements can commonly include a mixture of Vitamin E tocols having a variety of particular structures.

[0109] In some embodiments, a Vitamin E agent may include one or more sterocenters. Typically, the term “Vitamin E agent” is understood in encompass stereoisomeric (e.g., enantiomeric or diastereomeric) forms of a particular Vitamin E agent, as well as all geometric or conformational isomeric forms. For example, the R and S configurations of each stereocenter of a Vitamin E agent are contemplated in accordance with the presentdisclosure. In some embodiments, a relevant Vitamin E agent may be utilized as a single stereochemical or geometric isomer; alternatively, in some embodiments, a Vitamin E agent may be utilized as an enantiomeric, diastereomeric, or geometric (or conformational) mixture..

[0110] For example, in some embodiments a Vitamin E agent can be or include a stereoisomer of a Vitamin E tocopherol or tocotrienol. Tocopherol molecules have at leastthree stereocenters, at C-2, C-4' and C-8', making possible at least eight stereoisomers. For instance, in some embodiments, a Vitamin E agent can be or include a stereoisomer of a- tocopherol selected from RRR, RRS, RSS, SSS, RSR, SRS, SRR, and SSR. In various embodiments, the naturally occurring RRR stereoisomer is utilized. In some embodiments, a Vitamin E agent includes a mixture of one or more stereoisomers, e.g., one or more a- tocopherol stereoisomers (e.g., 1, 2, 3, 4, 5, 6, 7, or 8 stereoisomers). Natural tocopherols occur in the RRR-configuration while synthetic forms can include eight different stereoisomers and is called all-rac-alpha-tocopherol. Tocotrienols possess only the stereocenter at C-2 and naturally occurring tocotrienols are exclusively in the 2R,3’E,7’E configuration.

[0111] In some embodiments, a Vitamin E agent may be or comprise a derivatives and / or analog of a reference Vitamin E tocols (e.g., of a naturally occurring Vitamin E tocol). Those skilled in the art will appreciate that various Vitamin E tocols are amenable to modification, e.g., at the chroman moiety of a Vitamin E tocols. Examples of derivatives of Vitamin E tocols include 2-, 5- or 6-substituted chroman derivatives. Vitamin E tocols can be derivatized by conjugation with an agent such as a peg moiety (pegylation) or an acid (e.g., hyaluronic acid).

[0112] In some embodiments, a Vitamin E agent can be or include a prodrug of a Vitamin E tocol, e.g., an ester of a Vitamin E tocol (e.g., an ester of acetic acid, succinic acid, or nicotinic acid). In certain embodiments, an ester of a Vitamin E agent is prepared from a phenol form of a Vitamin E agent using conventional methods known in the art. Tocopheryl esters (e.g., alpha-tocopheryl acetate, tocopheryl succinate, tocopheryl nicotinate, tocopheryl linolate, alpha-tocopheryl phosphates, etc.) can demonstrate decreased susceptibility to oxidation. Tocopheryl esters can be de-esterified in the gut (e.g., by the enzyme esterase) and absorbed as free tocopherol. In some embodiments, a Vitamin E agent can be or include an unesterified Vitamin E tocol. In various embodiments, free and esterified Vitamin E tocols are understood to have comparable bioavailability.

[0113] In some embodiments, Vitamin E agents are absorbed, e.g., by the human body. The present disclosure includes both liquid and solid forms of Vitamin E. Exemplary solid forms of Vitamin E can be white to tan-white granular powders. Liquid forms of Vitamin E can be insoluble in water, soluble in alcohol, and / or miscible (e.g., with ether, acetone, vegetable oils, and / or chloroform). Various liquid forms of Vitamin E can be clear, yellow to brownish red, and / or viscous oils. The present disclosure provides Vitamin Eagents that are fat soluble. For example, in various embodiments, solid forms such as alpha tocopheryl acid succinate areinsoluble in water but soluble in fat (e.g., vegetable oil), and in various embodiments can coat crystallized semaglutide polypeptide and / or semaglutide polypeptide particles.

[0114] In some embodiments, a provided composition may include a Vitamin E agent as a pharmaceutically acceptable carrier; in various embodiments such Vitamin E agent may also provide nutritional supplementation (which does not necessarily alter its status or description as a carrier). Alternatively or additionally, in some embodiments, a provided composition may include an oil component as a pharmaceutically acceptable carrier. Thus, in various embodiments, the present disclosure provides compositions that include a pharmaceutically acceptable carrier comprising a Vitamin E agent (e.g., a fat soluble Vitamin E agent) and an oil.

[0115] In various embodiments, provided composition may includea a Vitamin E agent that is naturally present in or has been added to an oil component included in the composition. In some embodiments, an oil component is a plant-derived oil, optionally wherein the plant derived oil is selected from wheat germ oil, hazelnut oil, canola / rapeseed oil, sunflower oil, safflower oil, almond oil, grapeseed oil, sunflower seed kernels, almonds, almond butter, wheat germ, canola oil, palm oil, peanut oil, margarine, tub, hazelnuts, corn oil, olive oil, soybean oil, pine nuts, peanut butter, and peanuts. Exemplary concentrations of Vitamin E agent in certain such compositions are provided in Table 1. These provided concentrations are merely exemplary and in some instances reflect the amount of Vitamin E agent typically or naturally present, while the present disclosure includes that the amount of Vitamin E agent in oil compositions can be modified, e.g., by addition of Vitamin E agent to achieve a Vitamin E agent amount or concentration disclosed herein for use in a provided composition.

[0116] In some embodiments, a polypeptide composition (e.g., a carrier polypeptide composition, a semaglutide polypeptide composition) includes about 1%, 1.5%, 2%, 2.5%, 3%, 3.5%, 4%, 4.5%, 5%, 5.5%, or 6% weight per weight (w / w) Vitamin E agent in the composition.

[0117] In some embodiments, a polypeptide composition (e.g., a carrier polypeptide composition, a semaglutide polypeptide composition) includes at least 1%, 1.5%, 2%, 2.5%, 3%, or 3.5% weight per weight (w / w) Vitamin E agent in the composition.

[0118] In some embodiments, a polypeptide composition (e.g., a carrier polypeptide composition, a semaglutide polypeptide composition) includes about 3.5% w / w Vitamin E agent in the composition.

[0119] In some embodiments, a polypeptide composition (e.g., a carrier polypeptide composition, a semaglutide polypeptide composition) includes a coating or shell that comprises a Vitamin E agent.Table 1: Oils and Vitamin E Agent ConcentrationsPHARMACEUTICALLY ACCEPTABLE CARRIERS

[0120] Formulations provided herein can include a pharmaceutically acceptable carrier. As used herein, the term “pharmaceutically acceptable carrier” refers to a pharmaceutically-acceptable material, composition, or vehicle, such as a liquid or solid filler, diluent, excipient, or solvent encapsulating material, that facilitates formulation of an agent (e.g., a pharmaceutical agent), modifies bioavailability of an agent, or facilitates transport of an agent from one organ or portion of a subject to another. Some examples of materials which can serve as pharmaceutically-acceptable carriers include: Vitamin E agents, sugars, such as lactose, glucose and sucrose; starches, such as com starch and potato starch; cellulose, and its derivatives, such as sodium carboxymethyl cellulose, ethyl cellulose and cellulose acetate; powdered tragacanth; malt; gelatin; talc; excipients; oils, such as peanut oil, cottonseed oil, virgin coconut oil, almond oil, wheatgerm oil, any edible oil, safflower oil, sesame oil, olive oil, corn oil and soybean oil; glycols, such as propylene glycol; polyols, such as glycerin, sorbitol, mannitol and polyethylene glycol; esters, such as ethyl oleate and ethyl laurate; agar; buffering agents, such as magnesium hydroxide and aluminum hydroxide; alginic acid; pyrogen-free water; isotonic saline; Ringer’s solution; ethyl alcohol; pH buffered solutions; polyesters, polycarbonates and / or poly anhydrides; food ingredients, e.g., chocolate, cocoa, milk, milk product, natural dye, artificial dye, gum base, flavors, sweeteners, gelatin, starch, syrup, citric acid, and other non-toxic compatible substances employed in pharmaceutical formulations.

[0121] In some embodiments, a pharmaceutically acceptable carrier of a provided semaglutide polypeptide formulation comprises a Vitamin E agent. In some embodiments, a pharmaceutically acceptable carrier of a provided semaglutide polypeptide formulation is a Vitamin E agent. In some embodiments, the present disclosure provides formulations that comprise (1) a semaglutide polypeptide composition (e.g., a crystalline semaglutide polypeptide composition which may, in some embodiments, be a powder); and (2) one or more pharmaceutically acceptable carriers or excipients comprising Vitamin E. In some embodiments, provided oral formulations include (3) a carrier polypeptide composition (e.g., a crystalline carrier polypeptide composition).

[0122] Excipients can include a non-therapeutic agent that may be included in a pharmaceutical composition, for example to provide or contribute to a desired consistency or stabilizing effect. In some embodiments, suitable pharmaceutical excipients may include, for example, starch, glucose, lactose, sucrose, gelatin, malt, rice, flour, chalk, silica gel, sodium stearate, glycerol monostearate, talc, sodium chloride, dried skim milk, cocoa butter,suppository waxes, glycerol, propylene, glycol, water, ethanol, or the like. In various embodiments, one or more pharmaceutically acceptable carriers are selected from the group consisting of aggregation-reducing agents, sugars or sugar alcohols, polysaccharides, stabilizers, hyaluronidase, buffering agents, preservatives, carriers, antioxidants, chelating agents, natural or synthetic polymers, cryoprotectants, lyoprotectants, surfactants, bulking agents, acidifying agents, ingredients to reduce injection site discomfort, antifoaming agents, alkalizing agents, vehicles, aggregation inhibitors, permeation enhancers, muco bioadhesive agents, solubilizing agents, tonicity modifiers, and stabilizing agents and combinations thereof.

[0123] In various embodiments, aggregation-reducing agents can include one or more of nicotinic acid, caffeine citrate, caffeine nicotinate, caffeine, octyl- P-D-glucopyranoside, and n-dodecyl-P-D-maltoside and optionally in combination with one or more of arginine, tryptophan, histidine, proline, cysteine, methionine, P-alanine, Potassium Glutamate, Arginine Ethylester, lysine, aspartic acid, glutamic acid, glycine, DTPA (diethylenetriaminepentaacetic acid), EGTA(aminopolycarboxylic acid), EDTA (Ethylenediaminetetraacetic acid), hydroxy propyl beta (HP-Beta) cyclodextrins, hydroxy propyl gamma (HP-Gamma) cyclodextrins, sulfo-butyl ether (SBE) cyclodextrins, TMAO (trimethylamine N-oxide), trehalose, ethylene glycol, betaine, xylitol, sorbitol, 6-(N-(7- nitrobenz-2-oxa-l,3-diazol-4-yl)amino)hexanoic acid (NBD-X), methyl acetyl phosphate (MAP), citraconic anhydride, pyrophosphate, citrate, and combinations thereof.

[0124] In various embodiments, tonicity modifiers can include one or more of arginine, cysteine, histidine, glycine, sodium chloride, potassium chloride, sodium citrate, saccharides such as sucrose, glucose, dextrose, glycerin or mannitol, and combinations thereof.

[0125] In various embodiments, antioxidants can include one or more of glycine, lysine, EDTA, DTPA, sorbitol, mannitol, ascorbic acid, ascorbyl palmitate, butylated hydroxyanisole, butylated hydroxytoluene, hypopho sphorous acid, monothioglycerol, propyl gallate, sodium bisulfite, sodium formaldehyde sulfoxylate, sodium metabisulfite, sodium thiosulfate, sufur dioxide, tocopherol, and combinations thereof.

[0126] In various embodiments, lyoprotectants can include one or more of sucrose, lactose, trehalose, dextran, erythritol, arabitol, xylitol, sorbitol, maltose, lactulose, maltulose, glucitol, maltitol, lactitol, isomaltulose and mannitol; amino acids, such as arginine or histidine or proline or glycine; lyotropic salts, such as magnesium sulfate; propylene glycol,glycerol, poly(ethylene glycol), or polypropylene glycol); gelatin, dextrins, modified starch, carboxymethyl cellulose, and combinations thereof.

[0127] In various embodiments, permeation enhancer(s) can include one or more of bile salts, e.g., tri-hydroxy salts sodium cholate, sodium glyco-cholate, sodium taurocholate and di-hydroxy salt, sodium deoxy cholate, sodium glyco-deoxy Cholate, sodium tauro- deoxy cholate; fatty acids, their salt and esters, e.g., oleic acid, lauric acid, cod liver oil extract, sodium laurate, sodium caprate, glyceryl monostearate, di-ethylene glycol mono ethyl ether and various sucrose fatty acid esters, medium-chain fatty acid glycerides, polycaprolactoneomega-3 fatty acids, lecithin (phosphatidylcholine), lysophosphatidylcholine; surfactants, e.g., sodium dodecyl (lauryl) sulphate, poly sorbates (polysorbate 80), laureths, brijs and benzalkonium chloride; complexing agents, e.g., cyclodextrins, dextran sulphate, dextran sulphate, sodium edetate; complexing agents, e.g., cyclodextrins, dextran sulphate, dextran sulphate, sodium edetate, co-solvents, e.g., ethanol and propylene glycol, combination of 1% oleic acid and 5% / 10% polyethylene glycol 200, 2% glyceryl mono laurate and 40% alcohol, sodium caprate and alcohol or propylene glycol, 10% lauric acid in propylene glycol, polyoxyethylene, 2,3-lauryl ether, menthol, sodium caprate, sodium caprylate, sodium glycodeoxy cholate, glycol; polysaccharides, e.g., chitosan and chitosan glutamate; and others such as, aprotinin, benzalkonium chloride, cetylpyridinium chloride, cetyltrimethyl ammonium bromide, sodium salicylate, lysophosphatidylcholine, methoxysalicylate, methyloleate , sodium edta, sulfoxides, various alkyl glycosides, ethylene-diamide tetra acetic acid (edta), tartaric acid; lyotropic salts, such as magnesium sulfate; propylene glycol, glycerol, poly(ethylene glycol), or polypropylene glycol); gelatin, dextrins, modified starch, carboxymethyl cellulose, and combinations thereof.

[0128] In various embodiments, absorption enhancer(s) can include surfactants, cholesterol, glycerides, salicylates, bile salts, chelating agents, sodium caprate, a salt of capric acid and other includes N-(5-chlorosalicylol)-8-aminocaprylic acid (5-CNAC), 4-((4-chloro- 2-hydroxybenzoyl))-amino) butanoic acid (4-CNAB) and N-(8-(2-hydroxybenzoyl))-amino) caprylic acid, also known as salcaprozate sodium (SNAC, caprylic acid, C8, castor oil, medium chain, acyl carnitine, EDTA, glyceryl monolaurate, bovine P-casein, tocopherol succinate glycol chitosan conjugates, lecithins, glyceryl monostearate (GMS), chitosan and alginate, PLGA, silica, stearic acid, oleic acid, hydrogenated castor oil, and glyceryl trimyristate, etoposide phosphate (Vepesid®), sulindac (Clinoril®), enalapril maleate (Vasotec®), ramipril (Altace®), olmesartan medoxomil (Benicar®), valacyclovir (Valtrex®),midodrine (Amatine®), gabapentin enacarbil (Horizant®), sulfasalazine (Azulfidine®), and combinations thereof.

[0129] In various embodiments, muco bioadhesive agent(s) can include sucrose, lactose, trehalose, dextran, erythritol, arabitol, xylitol, sorbitol, maltose, lactulose, maltulose, glucitol, maltitol, lactitol, isomaltulose and mannitol; amino acids, such as arginine or histidine or proline or glycine; lyotropic salts, such as magnesium sulfate; propylene glycol, glycerol, poly(ethylene glycol), or polypropylene glycol); gelatin, dextrins, modified starch, carboxymethyl cellulose, and combinations thereof, mucoadhesive system such as from naturae, e.g., gelatin, agarose, chitosan, hyaluronic acid and synthetic polymers, e.g., polyvinylpyrolidone (PVP), polycrylates, polyvinyl alcohol, sodium carboxymethyl cellulose (SCMC) and pectin, all anionic-type polymers, chitosan (cationic type), and hydroxypropyl methylcellulose (HPMC) as a nonionic polymer, polyacrylic acid (PAA) derivatives (CP934, CP940, PCP), 15% CMC and 35% CP, copolymers of acrylic acid and poly(ethylene glycol) monomethylether monomethacrylate (PEGMM), eudragitl NE40D is a neutral poly(ethylacrlate methylmethacrylate, hydrophilic polymers, e.g., methocel K4M, methocel K15M, SCMC 400, Cekol 700, Cekol 10000, CP934P, CP971P and CP974P, carboxyvinyl polymer and triethanolamine, HPC(hydroxy propyl celluose), CP (carbopol 934P), carbopol (CP) EX-55 CMC (sodium carboxymethyl cellulose), HPMC (hydroxy propyl methyl cellulose), HEC (hydroxy ethyl cellulose), PIP [poly(isoprene)], PIB [poly (Isobutylene)], xanthum gum, locust bean gum, pectin, polycarbophil, benzyl esters, hydroxyethylcellulose, poly(acrylic acid), poly(acrylic acid-co-acrylamide), poly(acrylic acid-co-methyl methacrylate), poly(acrylic acid-co-butylacrylate), HEMA copolymerized with Polymeg® (poly tetramethylene glycol), Cydot® (bioadhesive polymeric blend of CP and PIB), formulation consisting of PVP, cetylpyridinium chloride (as stabilizer), chitosan chloride, polyethylene oxide, polymethylvinylether / maleic anhydride (PME / MA), and tragacanth, poly ethyleneglycol monomethylether monomethacrylate, drum dried waxy maize starch (DDWM), carbopol 974P, and sodium stearylfumarate, and cellulose derivatives; hyderogels-acrylic acid (polar) and butyl acrylate (apolar), and combinations thereof.

[0130] In various embodiments, pharmaceutically acceptable carriers expressly exclude a pharmaceutically active agent. In various embodiments, pharmaceutically acceptable carriers expressly exclude one or more, or all, biologically active polypeptides(e.g., those polypeptides sufficiently active to be considered an “active” agent in a drug product.)CRYSTALLIZED POLYPEPTIDES

[0131] The present disclosure includes oral formulations that include a polypeptide component that is or comprises one or more polypeptides. Typically, a polypeptide component will be or comprise a polypeptide in a crystalline (e.g., crystallized) form or in an amorphous form. Thus, in many embodiments, provide formulations include a semaglutide polypeptide (e.g., present in a polypeptide composition or a crystallized polypeptide composition) and a pharmaceutically acceptable carrier.

[0132] Those skilled in the art will appreciate that pharmaceutical agents, e.g., polypeptides (e.g., those in a carrier polypeptide composition, those in a polypeptide composition), often can exist in a variety of solid forms, including polymorph, solvate, hydrate, salt, co-crystal and amorphous forms. In certain embodiments, provided formulations include a polypeptide component (e.g., polypeptide in a carrier polypeptide composition, polypeptide in a polypeptide composition) in which at least one, and in some embodiments all, polypeptide(s) is in such a form. In some embodiments, a polypeptide component consists of crystalline polypeptide(s). In some embodiments, a polypeptide component is amorphous (e.g., consists of amorphous polypeptide(s)).

[0133] In some embodiments, a carrier polypeptide composition of a provided formulation is crystalline. In some embodiments, a semaglutide polypeptide composition of a provided formulation is crystalline. In some embodiments, a semaglutide polypeptide embedded in a carrier polypeptide composition of a provided formulation is crystalline. In some embodiments, a semaglutide polypeptide embedded in a carrier polypeptide composition, together with such carrier polypeptide composition form a co-crystal.

[0134] In some embodiments, a carrier polypeptide composition of a provided formulation is crystalline. In some embodiments, a semaglutide polypeptide composition of a provided formulation is crysattline (e.g., crystallized). In some embodiments, a semaglutide polypeptide embedded in a carrier polypeptide composition of a provided formulation is crystalline.

[0135] Amorphous polypeptide compositions can include compositions in which carrier polypeptide molecules are disordered or essentially disordered. Amorphous carrier polypeptides can lack or essentially lack long-range order of the positions of the atoms. In various embodiments, an amorphous carrier polypeptide can be more soluble than a crystallized form of the same polypeptide. In certain embodiments, an amorphous carrier polypeptide is a composition that has not been crystallized and / or has not been processed according to a method of crystallization. In certain embodiments, an amorphous carrierpolypeptide composition can include short-range order, residual crystallinity, polymorphic states, and regions of different density, none of which necessarily constitute long-range order. In various embodiments an amorphous carrier polypeptide composition can include a fraction of crystallized polypeptide, e.g., a fraction of crystallized polypeptide that is less than 20% total polypeptide (e.g., less than 20%, 15%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, 0.5%, or 0.1% crystallized polypeptide) by mass, volume, or mols. Techniques for determining the degree of crystallinity include XRD, DSC, solution calorimetry, water sorption, isothermal calorimetry, and thermally stimulated current (TSC). In various embodiments, an amorphous carrier polypeptide compositions does not diffract X-rays in a coherent manner and / or powder X-ray diffraction patterns are broad halos with no or very few characteristic peaks.

[0136] In various embodiments, a polypeptide composition (e.g., a carrier polypeptide composition, a semaglutide polypeptide composition) includes an average and / or maximum particle size of less than 25 microns, e.g., less than 20, 15, 10, 5, 4, 3, 2, 1, 0.5, or 0.1 microns, optionally wherein the particles have an average and / or maximum particle size that is between 0.1 microns and 0.5, 1, 2, 3, 4, 5, 10, 15, 20, or 25 microns or wherein the particles of polypeptide have an average and / or maximum particle size that is between 0.1 micron and 0.1, 0.5, 1, 2, 3, 4, 5, 10, 15, 20, or 25 microns. In various embodiments, particles of a polypeptide composition are microparticles or nanoparticles of polypeptide.

[0137] In some embodiments, polypeptide compositions (e.g., a carrier polypeptide composition, a semaglutide polypeptide composition) include compositions that include a high proportion of amorphous and / or non-crystallized polypeptide (e.g., of a polypeptide characterized by a particular amino acid sequence) relative to other agents or types of agents. In various embodiments, a polypeptide composition (e.g., a carrier polypeptide composition, a polypeptide composition) includes at least 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97% ,98%, 99%, or 100% amorphous and / or non-crystallized polypeptide (e.g., of a polypeptide characterized by a particular amino acid sequence) by weight, mole ratio, or volume of the composition or of polypeptide present in the composition. In various embodiments, a polypeptide composition (e.g., a carrier polypeptide composition, a semaglutide polypeptide composition) can be characterized by an amount of amorphous and / or non-crystallized polypeptide (e.g., of a polypeptide characterized by a particular amino acid sequence) that is in a range between a lower bound of 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 75%, 80%, 85%, or 90% by weight, mole ratio, or volume of the composition or ofpolypeptide present in the composition and an upper bound of 10%, 20%, 30%, 40%, 50%, 60%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97% ,98%, 99%, or 100% by weight, mole ratio, or volume of the composition or of polypeptide present in the composition.

[0138] In various embodiments, a polypeptide (e.g., a carrier polypeptide composition, a semaglutide polypeptide composition) composition is free, or substantially free, of non-polypeptide agents (and / or of agents other than a polypeptide), e.g., where the polypeptide composition includes no more than 1%, 2%, 3%, 4%, 5%, 10%, 15%, 20%, 25%, or 30% of non-polypeptide agents (and / or of agents other than a polypeptide) by weight, by mole ratio, or by volume. In various embodiments, a polypeptide composition (e.g., a carrier polypeptide composition, a polypeptide composition) includes no more than 1%, 2%, 3%, 4%, 5%, 10%, 15%, 20%, 25%, 30%, 40%, 50%, 60%, 70%, 75%, 80%, 85%, 90%, or 95% of non-polypeptide agents (and / or of agents other than a polypeptide) by weight, by mole ratio, or by volume. In various embodiments, a polypeptide composition (e.g., a carrier polypeptide composition, a semaglutide polypeptide composition) includes particles of one or more particular polypeptides and is free or substantially free of other polypeptide agents (optionally including crystallized form(s) of the one or more particular polypeptides), e.g., where the polypeptide composition includes no more than 1%, 2%, 3%, 4%, 5%, 10%, 15%, 20%, 25%, or 30% of other polypeptide agents by weight, by mole ratio, or by volume. In various embodiments, a polypeptide composition (e.g., a carrier polypeptide composition, a polypeptide composition) includes no more than 1%, 2%, 3%, 4%, 5%, 10%, 15%, 20%, 25%, 30%, 40%, 50%, 60%, 70%, 75%, 80%, 85%, 90%, or 95% of other polypeptide agents by weight, by mole ratio, or by volume. In various embodiments, an polypeptide composition (e.g., a carrier polypeptide composition, a polypeptide composition) includes particles of one or more particular polypeptides and is free or substantially free of other agents (optionally including crystallized form(s) of the one or more particular polypeptides), e.g., where the amorphous polypeptide composition includes no more than 1%, 2%, 3%, 4%, 5%, 10%, 15%, 20%, 25%, or 30% of other agents by weight, by mole ratio, or by volume. In various embodiments, a polypeptide composition (e.g., a carrier polypeptide composition, a polypeptide composition) includes no more than 1%, 2%, 3%, 4%, 5%, 10%, 15%, 20%, 25%, 30%, 40%, 50%, 60%, 70%, 75%, 80%, 85%, 90%, or 95% of other agents by weight, by mole ratio, or by volume.

[0139] In various embodiments, a polypeptide composition (e.g., a carrier polypeptide composition, a polypeptide composition) includes particles of one or more particular polypeptides where the particles of the one or more particular polypeptides are free, orsubstantially free, of non-polypeptide agents, e.g., where the particles include no more than 1%, 2%, 3%, 4%, 5%, 10%, 15%, 20%, 25%, or 30% of non-polypeptide agents by weight, by mole ratio, or by volume. In various embodiments, a polypeptide (e.g., a carrier polypeptide composition, a polypeptide composition) composition includes particles of one or more particular polypeptides where the particles of the one or more particular polypeptides include no more than 1%, 2%, 3%, 4%, 5%, 10%, 15%, 20%, 25%, 30%, 40%, 50%, 60%, 70%, 75%, 80%, 85%, 90%, or 95% of non-polypeptide agents by weight, by mole ratio, or by volume. In various embodiments, a polypeptide composition (e.g., a carrier polypeptide composition, a polypeptide composition) includes particles of one or more particular polypeptides where the particles of the one or more particular polypeptides are free, or substantially free, of other polypeptides, e.g., where the particles of the one or more particular polypeptides include no more than 1%, 2%, 3%, 4%, 5%, 10%, 15%, 20%, 25%, or 30% of other polypeptides by weight, by mole ratio, or by volume. In various embodiments, a polypeptide composition (e.g., a carrier polypeptide composition, a polypeptide composition) includes particles of one or more particular polypeptides where the particles of the one or more particular polypeptides include no more than 1%, 2%, 3%, 4%, 5%, 10%, 15%, 20%, 25%, 30%, 40%, 50%, 60%, 70%, 75%, 80%, 85%, 90%, or 95% of other polypeptides by weight, by mole ratio, or by volume. In various embodiments, a polypeptide composition (e.g., a carrier polypeptide composition, a polypeptide composition) includes particles of one or more particular polypeptides where the particles of the one or more particular polypeptides are free, or substantially free, of other agents (optionally including non-crystallized form(s) of the one or more particular polypeptides), e.g., where the particles of the one or more particular polypeptides include no more than 1%, 2%, 3%, 4%, 5%, 10%, 15%, 20%, 25%, or 30% of other agents by weight, by mole ratio, or by volume. In various embodiments, a polypeptide composition (e.g., a carrier polypeptide composition, a polypeptide composition) includes particles of one or more particular polypeptides where the particles of the one or more particular polypeptides include no more than 1%, 2%, 3%, 4%, 5%, 10%, 15%, 20%, 25%, 30%, 40%, 50%, 60%, 70%, 75%, 80%, 85%, 90%, or 95% of other agents by weight, by mole ratio, or by volume.

[0140] In some embodiments, processes for production of crystallized polypeptide compositions can include one or more of, for example, molecular quenching of melts, rapid precipitation by antisolvent addition, freeze-drying, spray-drying, spray-freeze-drying, precipitation in supercritical fluids, solid-dispersion, and solid-state chemical reactions(degradation) of crystalline precursors. For example, freeze drying of a protein / PEG blendsolution and subsequent removal of PEG from the matrix has proven to yield precipitated protein particles in amorphous form. Processes that introduce mechanical or chemical stress (grinding, milling, and wet granulation) can render crystalline materials fully or partially amorphous.

[0141] In some embodiments, a polypeptide composition can be a hydrated form or prepared from a hydrated form. Hydrated forms can include an alcohol (e.g., ethanol). In some embodiments, a polypeptide composition is a solvated form or prepared form a solvated form. Exemplary solvents can include, for example, an acidic solvent or an organic solvent. In some embodiments, a solvent can include DMSO, DMF, acetic acid, acetonitrile, methanol, propanol, isopropanol, acetone, anisole, 1 -butanol, 2-butanol, butyl acetate, fer / - butylmethyl ether, cumene, dimethyl sulfoxide, ethanol, ethyl acetate, ethyl ether, ethyl formate, formic acid, heptane, isobutyl acetate, isopropyl acetate, methyl acetate, 3 -methyl- 1 -butanol, methylethyl ketone, methylisobutyl ketone, 2-methyl-l-propanol, pentane, 1- pentanol, 1 -propanol, 2-propanol, propyl acetate, tetrahydrofuran, ethanol, and / or water.Certain methods of preparing crystallized polypeptide compositions can include removal of a solvent by rapid solvent evaporation from a solvated form, spray drying, roller drying, solvent precipitation, or freeze drying. In some embodiments, a polypeptide composition includes a cation such as a 2+ charged cation (e.g., Ba2+, Ca2+, Cr2+, Co2+, Cu2+, Fe2+, Pb2+, Mg2+, Mn2+, Ni2+, Sr2+, Sn2+, or Zn2+).

[0142] In some embodiments, crystal formation can include assembly of noncrystalline solid agents into a crystalline solid form. In some embodiments, crystal formation can utilize various molecular interactions, including, e.g., hydrogen bonding, p stacking, and Vander Waals Forces. Hydrogen bond formation is often responsible for intermolecular interactions in molecular solids. Crystallization is typically considered with respect to small molecule agents, and rarely considered with respect to polypeptides. Polypeptides typically lack well-defined conformation in solution. In some embodiments a polypeptide composition is a crsyallized polypeptide composition. In certain embodiments, a crystallized polypeptide composition is a powder form that includes crystals of polypeptide.

[0143] In various embodiments, a crystallized polypeptide composition (e.g., a carrier polypeptide composition, a polypeptide composition) includes crystals of polypeptide having an average and / or maximum particle size of less than 25 microns, e.g., less than 20, 15, 10, 5, 4, 3, 2, 1, 0.5, or 0.1 microns, optionally wherein the crystals of polypeptide have an average and / or maximum particle size that is between 0.1 microns and 0.5, 1, 2, 3, 4, 5, 10, 15, 20, or 25 microns or wherein the crystals of polypeptide have an average and / or maximum particlesize that is between 0.1 micron and 0.1, 0.5, 1, 2, 3, 4, 5, 10, 15, 20, or 25 microns. In various embodiments, crystals of a crystallized polypeptide composition (e.g., a carrier polypeptide composition, a polypeptide composition) are microcrystals or nanocrystals of polypeptide.

[0144] In some embodiments, crystallized polypeptide compositions include compositions that include a high proportion of crystallized polypeptide (e.g., of a polypeptide characterized by a particular amino acid sequence) relative to other agents or types of agents. In various embodiments, a crystallized polypeptide composition includes at least 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 75%, 80%, 85%, 90%, 95%, or 100% crystallized polypeptide (e.g., of a polypeptide characterized by a particular amino acid sequence) by weight, by mole ratio, or by volume of the composition or of polypeptide present in the composition. In various embodiments, a crystallized polypeptide composition can be characterized by an amount of crystallized polypeptide (e.g., of a polypeptide characterized by a particular amino acid sequence) that is between that is between a lower bound of 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 75%, 80%, 85%, or 90% by weight, mole ratio, or volume of the composition or of polypeptide present in the composition and an upper bound of 10%, 20%, 30%, 40%, 50%, 60%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97% ,98%, 99%, or 100% by weight, mole ratio, or volume of the composition or of polypeptide present in the composition.

[0145] In various embodiments, a crystallized polypeptide composition is free, or substantially free, of non-polypeptide agents e.g., where the crystallized polypeptide composition includes no more than 1%, 2%, 3%, 4%, 5%, 10%, 15%, 20%, 25%, or 30% of non-polypeptide agents by weight, by mole ratio, or by volume. In various embodiments, a crystallized polypeptide composition includes no more than 1%, 2%, 3%, 4%, 5%, 10%, 15%, 20%, 25%, 30%, 40%, 50%, 60%, 70%, 75%, 80%, 85%, 90%, or 95% of non- polypeptide agents by weight, by mole ratio, or by volume. In various embodiments, a crystallized polypeptide composition includes crystals of one or more particular polypeptides and is free or substantially free of other polypeptide agents (optionally including noncrystallized form(s) of the one or more particular polypeptides), e.g., where the crystallized polypeptide composition includes no more than 1%, 2%, 3%, 4%, 5%, 10%, 15%, 20%, 25%, or 30% of other polypeptide agents by weight, by mole ratio, or by volume. In various embodiments, a crystallized polypeptide composition includes crystals of one or more particular polypeptides and includes no more than 1%, 2%, 3%, 4%, 5%, 10%, 15%, 20%,25%, 30%, 40%, 50%, 60%, 70%, 75%, 80%, 85%, 90%, or 95% of other polypeptide agents by weight, by mole ratio, or by volume. In various embodiments, a crystallized polypeptide composition includes crystals of one or more particular polypeptides and is free or substantially free of other agents (optionally including non-crystallized form(s) of the one or more particular polypeptides), e.g., where the crystallized polypeptide composition includes no more than 1%, 2%, 3%, 4%, 5%, 10%, 15%, 20%, 25%, or 30% of other agents by weight, by mole ratio, or by volume. In various embodiments, a crystallized polypeptide composition includes crystals of one or more particular polypeptides and includes no more than 1%, 2%, 3%, 4%, 5%, 10%, 15%, 20%, 25%, 30%, 40%, 50%, 60%, 70%, 75%, 80%, 85%, 90%, or 95% of other agents by weight, by mole ratio, or by volume.

[0146] In various embodiments, a crystallized polypeptide composition includes crystals of one or more particular polypeptides where the crystals of the one or more particular polypeptides are free, or substantially free, of non-polypeptide agents, e.g., where the crystals include no more than 1%, 2%, 3%, 4%, 5%, 10%, 15%, 20%, 25%, or 30% of non-polypeptide agents by weight, by mole ratio, or by volume. In various embodiments, a crystallized polypeptide composition includes crystals of one or more particular polypeptides where the crystals include no more than 1%, 2%, 3%, 4%, 5%, 10%, 15%, 20%, 25%, 30%, 40%, 50%, 60%, 70%, 75%, 80%, 85%, 90%, or 95% of non-polypeptide agents by weight, by mole ratio, or by volume. In various embodiments, a crystallized polypeptide composition includes crystals of one or more particular polypeptides where the crystals of the one or more particular polypeptides are free, or substantially free, of other polypeptides, e.g., where the crystals of the one or more particular polypeptides include no more than 1%, 2%, 3%, 4%, 5%, 10%, 15%, 20%, 25%, or 30% of other polypeptides by weight, by mole ratio, or by volume. In various embodiments, a crystallized polypeptide composition includes crystals of one or more particular polypeptides where the crystals of the one or more particular polypeptides include no more than 1%, 2%, 3%, 4%, 5%, 10%, 15%, 20%, 25%, 30%, 40%, 50%, 60%, 70%, 75%, 80%, 85%, 90%, or 95% of other polypeptides by weight, by mole ratio, or by volume. In various embodiments, a crystallized polypeptide composition includes crystals of one or more particular polypeptides where the crystals of the one or more particular polypeptides are free, or substantially free, of other agents (optionally including non-crystallized form(s) of the one or more particular polypeptides), e.g., where the crystals of the one or more particular polypeptides include no more than 1%, 2%, 3%, 4%, 5%, 10%, 15%, 20%, 25%, or 30% of other agents by weight, by mole ratio, or by volume. In various embodiments, a crystallized polypeptide composition includes crystals of one or moreparticular polypeptides where the crystals of the one or more particular polypeptides include no more than 1%, 2%, 3%, 4%, 5%, 10%, 15%, 20%, 25%, 30%, 40%, 50%, 60%, 70%, 75%, 80%, 85%, 90%, or 95% of other agents by weight, by mole ratio, or by volume.

[0147] In some embodiments, polypeptide crystals can be prepared from a polypeptide sample. Polypeptide crystals can be prepared form a polypeptide sample that is substantially pure of other agents and / or contaminants. Polypeptide crystals can be prepared from an amorphous material, e.g., a lyophilized material that is amorphous or an amorphous solid that has been precipitated. In some embodiments, polypeptide crystals can be prepared from a mixture of amorphous and crystalline material, e.g., a lyophilized material that is a mixture of amorphous and crystalline material or a mixture of amorphous and crystalline solids that has been precipitated. In various embodiments, polypeptide crystals can be prepared form a polypeptide sample having a polypeptide concentration between 0.1 and 200 mg / mL, e.g., having a polypeptide concentration between a lower bound of 0.1, 1, 5, 10, 15, 20, 25, 50, 75, or 100 mg / mL and an upper bound of 15, 20, 25, 50, 75, 100, 125, 150, 175, or 200 mg / mL.

[0148] In some embodiments, means of polypeptide crystallization can include, without limitation, one or more of, evaporation, slow diffusion (e.g., vapor diffusion at ambient or low temperature), slow cooling, slurrying, hanging drop, sitting drop, seeded crystal development, and / or other crystallization methods known in the art. Those of skill in the art will appreciate that many crystallization methods are well understood and known in the art, such that polypeptide crystallization is generally straightforward. Those of skill in the art will further appreciate that polypeptide crystallization is especially straight forward where a particular crystal size is not required, where large crystals are not required, where a particular crystalline form is not required, and / or where perfectly regular crystals are not required, any or all of which can characterize various embodiments as described herein.

[0149] In some embodiments, crystalline polypeptides can be prepared by mixing a polypeptide in a suitable solvent (e.g., water) and then causing the polypeptide to return to the solid phase. For example, polypeptides can form crystals when precipitated from an aqueous solution (e.g., of ammonium sulfate). In certain exemplary embodiments, a polypeptide- saturated solution is prepared by increasing the concentration of the polypeptide in the solution. At maximal solubility, polypeptide precipitation can occur and the precipitant can be crystalline. Slow precipitation can produce small numbers of larger crystals while more rapid precipitation can produce very large numbers of small crystals, such that the rate of precipitation is therefore not critical to the production of crystals in general.

[0150] In some embodiments, evaporation (e.g., slow evaporation) is a common means of crystallizing polypeptides. Precipitation of a polypeptide can occur by allowing the solvent of a solution of polypeptide to evaporate (e.g., slowly evaporate) until the solution reaches saturation, thereby allowing polypeptide precipitation to occur.

[0151] In some embodiments, cooling (e.g., slow cooling) is another method of crystallizing polypeptides. Precipitation of a polypeptide can occur by allowing a solution of polypeptide to cool (e.g., slowly cool), thereby reducing the maximum solubility of the polypeptide in the solution and inducing precipitation to occur.

[0152] In some embodiments, vapor diffusion and batch methods are also commonly employed in polypeptide crystallization. In vapor diffusion, a drop containing a mixture of precipitant and unprecipitated polypeptide is sealed in a chamber with pure precipitant. Water vapor then diffuses out of the drop until the osmolarity of the drop and the precipitant are equal. The dehydration of the drop causes a slow concentration of both polypeptide and precipitant until equilibrium can be achieved, favoring crystallization. Vapor diffusion can be performed in either hanging-drop or sitting-drop format. A hanging-drop method can involve a drop of polypeptide solution placed on an inverted cover slip, which is then suspended above the reservoir. A sitting-drop method can position a drop on a pedestal that is separated from the reservoir. Both of these methods require sealing of the environment so that equilibration between the drop and reservoir can occur.

[0153] In some embodiments, a batch method relies on bringing a polypeptide directly into the nucleation zone by mixing polypeptide with the appropriate amount of precipitant. Various Examples provided herein include batch crystallization. Batch crystallization is different from continuous crystallization in that the withdrawal of crystal product for the batch system is made only once at the end of the batch run. Batch crystallization may also include the semibatch system, in which one or more feed solutions are added to the crystallizer at a constant or variable rate throughout all or part of the batch. In various embodiments, batch crystallization can vary in volume from, e.g., 1 microliter (e.g., in an Eppendorf tube) to a liter or more (e.g., thousands of liters). In various embodiments, no vapor diffusion method is involved. In various embodiments, no evaporation is involved. In various embodiments, batch crystallization includes slowly adding precipitating reagents (e.g., with stirring if necessary, depending on batch size).

[0154] Typically, seeds of crystallizing material can be added early in the batch process in order to improve reproducibility and product quality. When a desired amount of solid has been formed, slurry is typically transferred to a solid-liquid separation unit.

[0155] In some embodiments, dialysis is another method commonly employed in polypeptide crystallization. This technique utilizes diffusion and equilibration of precipitant molecules through a semi-permeable membrane as a means of gradually approaching the concentration at which the macromolecule crystallizes. Dialysis tubes can be used in the case of large amounts of polypeptide being available.

[0156] In some embodiments, microdialysis buttons, also known as Cambridge buttons, offer a convenient way to produce crystals from a small amount of sample. A polypeptide sample is placed inside a small chamber on top of the button and the sample is covered with a dialysis membrane of appropriate molecular weight cut-off. The apparatus is then immersed in a reservoir containing precipitant solution. Equilibration of precipitant molecules can occur through the membrane.

[0157] In some embodiments, free interface diffusion can also be used to crystallize polypeptide. This technique can include carefully layering precipitant solution on top of concentrated polypeptide solution in a capillary, the ends of which are then sealed with wax. Narrow diameter of the capillary minimizes mixing from natural convection in the system. Thus, precipitant and polypeptide slowly inter-diffuse and the system reaches the equilibrium by a phenomenon called counter-diffusion. When the solutions initially come into contact and diffusive mixing occurs, the region of the polypeptide solution in the neighborhood of the interface becomes supersaturated and ideal conditions for nuclei formation are created. As time proceeds, the two solutions inter-diffuse along the axis of the capillary and dilute each other, thus promoting the dissolution of the smaller nuclei and the growth of the larger ones. The achievement, by the free liquid diffusion, of transient nucleation conditions in most cases allows to obtain high quality crystals. Thus free interface diffusion can be view as a rational crystallization approach to minimize supersaturation and impurity levels at the crystal growth front and to ensure steadiness of both values. A variant of free interface diffusion method is referred to as liquid bridge method, in which method a drop of polypeptide sample and a drop of precipitant solution are placed in close proximity on a cover glass and connected by a thin liquid bridge. The liquid diffusion between the two droplets, sealed from air, may induce crystal growth.

[0158] In some instances, crystallization nucleation can be induced by use of a material such as a nucleating agent, nucleant, or seed. Nucleation can occur on the surface of a nucleating agent, nucleant, or seed, which induces a higher local concentration of macromolecules, lowers the energy barrier for nucleation and bypasses kinetic barriers ofspontaneous nucleation; a lower level of supers aturation can be required under such circumstances.

[0159] In some embodiments, pharmaceutical co-crystals can be crystalline materials comprised of a pharmaceutically active ingredient and one or more co-crystal formers (“coformers”), such that the active ingredient and coformers are together in the same crystal lattice. Co-crystals are distinguished from salts because unlike salts, the components that coexist in the co-crystal lattice with a defined stoichiometry interact nonionically. In addition, co-crystals differ from polymorphs, which are defined as including 1) single-component crystalline forms that have different arrangements or conformations of the molecules in the crystal lattice, 2) amorphous forms, and 3) multicomponent phases such as solvate and hydrate forms. Co-crystals are similar to solvates at least in that both contain more than one component in the lattice. In some embodiments, a crystallized polypeptide composition does not include co -crystallized polypeptide. In some embodiments, a crystallized polypeptide composition includes co-crystallized polypeptide.

[0160] In some embodiments, a polypeptide as described herein can be incorporated into a co-crystal during the process of crystallizing carrier polypeptide. In some embodiments, a polypeptide is a coformer during the crystallization of a carrier polypeptide. In some embodiments, a carrier polypeptide is a coformer during the crystallization of a polypeptide . In some embodiments, a carrier polypeptide and a polypeptide are together in the same crystal lattice. .

[0161] The present disclosure provides the recognition that a co-crystal comprising (1) a carrier polypeptide and (2) a polypeptide , can be formulated into a formulation as described herein. In some embodiments, a co-crystal is formulated into a truffle formulation. In some embodiments, a co-crystal is formulated into a globule formulation. In some embodiments, a co-crystal is formulated into a tablet formulation. In some embodiments, a co-crystal is formulated into a candy formulation. In some embodiments, a co-crystal is formulated into a formulation suitable for oral administration as described herein. In some embodiments, a co-crystal is formulated into a suspension formulation. In some embodiments, a co-crystal is formulated into a capsule formulation. In some embodiments, a co-crystal is formulated into a formulation optionally comprising a further pharmaceutically acceptable carrier. In some embodiments, a co-crystal is formulated into a formulation optionally comprising Vitamin E. In some embodiments, a co-crystal is formulated into a capsule formulation optionally comprising Vitamin E.

[0162] Those of skill in the art will appreciate that a number of crystallization conditions can be adjusted to increase or decrease efficiency and / or purity of crystallization. For instance, crystallization conditions that can be adjusted include solubilization systems (aqueous systems and / or organic solvent systems), pH, counterions, salts, temperature, excipients, coformers, and polypeptide concentration. Modulating and testing ranges for these factors is trivial for those of skill in the art. In the art of crystallization, it is considered typical to sample a wide variety of crystallization conditions. Moreover, as those of skill in the art will appreciate, the present disclosure does not necessarily require that the most efficient form of crystallization be identified, only that crystals can be formed.

[0163] In some embodiments, crystallized polypeptide compositions can include a plurality of crystallized polypeptides. In various embodiments, crystallized polypeptides can be characterized by high concentration, high purity, and / or high stability. Any suitable methods known in the art can be used to characterize provided crystallized polypeptide compositions, including but not limited to X-ray powder diffraction (XRPD), differential scanning calorimetry (DSC), thermogravimetric analysis (TGA), high-performance liquid chromatography (HPLC), liquid chromatography-mass spectrometry (LCMS), laser diffraction, hot stage microscopy, polarized light microscopy, and the like.

[0164] In various embodiments, a crystallized polypeptide composition (e.g., a carrier polypeptide composition, a polypeptide composition) is free, or substantially free, of one or more, or any, precipitation reagents, optionally wherein the precipitation reagents includes one or more of salts, organic solvents, or polymers, optionally where the salts can include a salt selected from ammonium sulfate, citrate salts, and cetyltrimethylammonium salts, the organic solvents can include an organic solvent selected from 2-methyl-2,4-pentanediol or 2- Methyl-2, 4-pentanediol, and / or the polymers can include a polymer that is a polyethylene glycol.

[0165] In various embodiments, a crystallized polypeptide composition (e.g., a carrier polypeptide composition, a polypeptide composition) includes polypeptide crystals that are free, or substantially free, of one or more, or any, precipitation reagents, optionally wherein the precipitation reagents includes one or more of salts, organic solvents, or polymers, optionally where the salts can include a salt selected from ammonium sulfate, citrate salts, and cetyltrimethylammonium salts, the organic solvents can include an organic solvent selected from 2-methyl-2, 4-pentanediol or 2-Methyl-2, 4-pentanediol, and / or the polymers can include a polymer that is a polyethylene glycol.

[0166] Except where otherwise specified, those of skill in the art will appreciate that references relating to crystallization in the art refer to crystallization of molecules that are not polypeptides, e.g., to crystallization of molecules that are small molecules, e.g., small molecule therapeutics.

[0167] In various applications, peptide crystallization can produce well-ordered crystals with generally uniform content. While the present disclosure includes polypeptide compositions that include well-ordered crystals, the present disclosure also specifically includes the recognition that formulations do not require crystals having a particular or consistent size or character. In various embodiments, crystallized polypeptide compositions include one or more crystalline forms of a polypeptide (e.g., one or more polymorphs or hydrates of a polypeptide).

[0168] Those of skill in the art will appreciate from the present disclosure that compositions and methods provided herein are useful for the delivery of a wide variety of polypeptides. Among other things, the present disclosure provides the recognition that a wide range of polypeptides with different characteristics (e.g., molecular weight, molecule size, hydrophilicity, lipophilicity, modification patterns, preparation methods, etc.) can be utilized in formulations and methods as described herein. Those skilled in the art will appreciate that polypeptides useful in formulations, compositions, and methods as described herein can be grouped into different categories in a variety of ways.

[0169] In some embodiments, a polypeptide is a stapled peptide. In some embodiments, a polypeptide is a cyclic peptide generated through cyclization. In some embodiments, a polypeptide is a peptide comprising bicycles through chemical linkage.

[0170] In some embodiments, a polypeptide is a hydrophobic peptide. In some embodiments, a polypeptide is a hydrophilic peptide.

[0171] In some embodiments, a polypeptide is a peptide comprising dual agonists.

[0172] In some embodiments, a polypeptide is generated through Phage display technology. In some embodiments, a polypeptide is generated through peptidomimetics approach. In some embodiments, a polypeptide is a polypeptide is generated through genetic engineering. In some embodiments, a polypeptide is generated through non-ribosomal method. In some embodiments, a polypeptide is rationally designed based on protein-protein interaction.

[0173] In some embodiments, a polypeptide is a polypeptide comprising sustained release properties.

[0174] In some embodiments, a polypeptide is able to establish intramolecular hydrogen bonding.

[0175] In some embodiments, a polypeptide is a peptide comprising pH dependent solubility and / or stability.

[0176] Those skilled in the art will appreciate that formulations or methods described herein provide surprisingly improved properties and such surprisingly improved properties are observed across a wide range of polypeptide formulations comprising polypeptides with different characteristics (e.g., molecular weight, molecule size, hydrophilicity, lipophilicity, modification patterns, preparation methods, etc.). For example, those of skill in the art will appreciate from the present disclosure that compositions and methods provided herein are advantageous at least in part because they deliver polypeptides to the bloodstream and / or plasma after oral administration, e.g., buccal / sublingual administration, e.g., with advantageous pharmacokinetic properties.

[0177] In some embodiments, a polypeptide amenable to formulation as described herein has a molecular weight that may be, for example, at least about 4 kDa, 25 kDa, 50 kDa, 100 kDa, 150 kDa, 200 kDa, 250 kDa, 300 kDa, 400 kDa, 500 kDa, 1,000 kDa, 2,000 kDa, or greater. In some embodiments, a polypeptide has a molecular weight within a range, for example, from about IDa to about 1,000 kDa, e.g., from about IkDa to about 5kDa, about 25 kDa to about 1,000 kDa, about 25 kDa to about 500 kDa, about 100 kDa to about 500 kDa, about 125 kDa to about 250 kDa, about 125 kDa to about 175 kDa, or about 150 kDa to about 300 kDa. In various embodiments, a polypeptide can have a molecular weight having a lower bound of e.g., about 100, 150, 200, 250, 300, 350, 400, 450, or 500 kDa and an upper abound of, e.g., about 100, 150, 200, 250, 300, 350, 400, 450, 500, 750, or 1,000 kDa.

[0178] In some embodiments, a polypeptide amenable to formulation as described herein has a molecular weight that may be, for example, between about 100 Da and 25 kDa, optionally wherein the molecular weight is between about 100 Da and about 1 kDa, about 100 Da and about 2 kDa, about 100 Da and about 3 kDa, about 100 Da and about 4 kDa, about 100 Da and about 5 kDa, about 100 Da and about 10 kDa, about 100 Da and about 15 kDa, or about 100 Da and about 20 kDa, In various embodiments, a polypeptide can have a molecular weight within a range having a lower bound of e.g., about 100 Da, 150 Da, 200 Da, 250 Da, 300 Da, 400 Da, 500 Da, 600 Da, 700 Da, 800 Da, 900 Da, 1 kDa, 2 kDa, 3 kDa, 4 kDa, or 5 kDa and an upper abound of, e.g., about 250 Da, 300 Da, 400 Da, 500 Da, 600 Da, 700 Da, 800 Da, 900 Da, 1 kDa, 2 kDa, 3 kDa, 4 kDa, 5 kDa, 10 kDa, 15 kDa, 20 kDa, 25 kDa, 50 kDa, 100 kDa, 150 kDa, 200 kDa, 300 kDa, 500 kDa, 800 kDa, 1,000 kDa, 1,500kDa, or 2,000 kDa, wherein the upper bound of the range is higher than the lower bound of the range.

[0179] In some embodiments, a polypeptide amenable to formulation as described herein has a molecular weight that may be, for example, between about 500 Da and 70 kDa. In some embodiments, a polypeptide has a molecular weight that may be, for example, between about 5 kDa and 2,000 kDa. In some embodiments, a polypeptide has a molecular weight that may be, for example, between about 10 kDa and 500 kDa. In some embodiments, a polypeptide has a molecular weight that may be, for example, between about 20 kDa and 500 kDa. In some embodiments, a polypeptide has a molecular weight that may be, for example, between about 100 kDa and 150 kDa. In some embodiments, a polypeptide has a molecular weight that may be, for example, between about 150 kDa and 200 kDa.

[0180] In some embodiments, a polypeptide amenable to formulation as described herein can include, e.g., about 3 to about 1,000 amino acids, e.g., about 10 to about 500, about 10 to about 250, about 10 to about 100, about 100 to about 1,000, about 100 to about 750, about 100 to about 500, about 100 to about 250, about 250 to about 1,000, about 250 to about 750, or about 250 to about 500 amino acids. In various embodiments, polypeptides can include a number of amino acids within a range having a lower bound of, e.g., about 3, 10, 20, 30, 40, 50, 75, 100, 150, 200, 250, 300, 350, 400, 450, or 500 amino acids and an upper abound of, e.g., about 100, 150, 200, 250, 300, 350, 400, 450, 500, 750, or 1,000 amino acids, the lower bound being smaller than the upper bound.

[0181] In various embodiments, a polypeptide amenable to formulation as described herein can be a therapeutic polypeptide, e.g., a semaglutide polypeptide, for administration to a subject in need thereof.

[0182] In various embodiments, polypeptides can be or include recombinant polypeptides, isolated or synthetic polypeptides, glycoproteins, phosphoproteins, lipoproteins, antibodies, pegylated polypeptides, therapeutic polypeptides, storage polypeptides, and combinations thereof.

[0183] The present disclosure includes analogs and modified forms of polypeptides disclosed herein, e.g., that include one or more pendant groups or modifications, e.g., pegylation, acetylation, amidation, lipidation, methylation, phosphorylation, glycosylation, glycation, sulfation, mannosylation, nitrosylation, acylation, palmitoylation, prenylation, or combinations thereof. In various particular embodiments, a polypeptide includes one or more pendant groups or modifications selected from pegylation and / or fatty acids. Those skilled in the art will appreciate that the modified polypeptides of the present disclosure encompasspolypeptides with non-natural amino acids, with altered amino acids, with altered intramolecular binding properties, with chemical modification of the backbone.

[0184] In some embodiments, a polypeptide is conjugated with another molecule, e.g., small molecules (e.g., fluorophores, chelating agents and therapeutics), oligosaccharides, lipids (e.g., ionizable cationic lipids), oligonucleotides, antibodies, etc. In some embodiments, a polypeptide is conjugated to a ligand that targets a site of therapeutic relevance. In some embodiments, a polypeptide is conjugated to a ligand that targets a cell surface receptor, e.g., carbohydrate receptors, lipoprotein receptors, transferring receptors, receptors involved cell adhesion, etc. In some embodiments, a polypeptide is conjugated to a cell penetrating peptide. In some embodiments, a polypeptide is conjugated to an albumin binding peptide. In some embodiments, a polypeptide is conjugated to a passive or active transport enhancers. In some embodiments, a polypeptide is conjugated to a mucoadhesive device.

[0185] In some embodiments, a polypeptide of the present disclosure is semaglutide.

[0186] The present disclosure further relates to engineered and / or biosimilar forms of therapeutic polypeptides, which such forms may, for example, show at least 80% sequence identity with their reference polypeptide, e.g., at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% identity with such reference polypeptide.

[0187] The present disclosure further includes the recognition that polypeptides, includingare generally not referred to as “small molecules” or “compounds” in the art. Accordingly, because small molecules and compounds can have very different physical and pharmacokinetic properties than polypeptides as disclosed herein, prior art disclosures relating to small molecules and compounds would not be understood as applicable by those of skill in the art to the formulations and methods relating to polypeptides as disclosed herein.

[0188] In various embodiments, a carrier polypeptide composition is or comprises, for example, selected from antibodies, contractile proteins, enzymes, hormonal proteins, structural proteins, storage proteins, and transport protein, or combinations thereof.

[0189] In some embodiments, a carrier polypeptide composition is or comprises, for example, selected from albumins, gluten, globulins, glutenin, prolamin, legumin, vicillin, glycinin, conglycinin, gliadins, myoglobulins, collagen, elastin, myosin, actin, tropomysin, troponin, alpha, beta and kappa caseins, alpha and beta zein, convicillin, gelatin, ovalbumin, sericin, fibroin, beta lactoglobulin, glutelins, helianthinin, immunoglobulins, ovomucoid, ovomucin lysozyme, or ovotransferrin, or a combination thereof.

[0190] In some embodiments, a carrier polypeptide composition is or comprises amylase.

[0191] In some embodiments, a carrier polypeptide composition is or comprises albumin.FORMULATIONS

[0192] As appreciated by those skilled in the art, in some aspects, a polypeptide formulation as described herein is in a form suitable for oral administration, e.g., buccal / sublingual administration. In some embodiments, a provided formulation is a truffle formulation. In some embodiments, a provided formulation is a tablet formulation. In some embodiments, a provided formulation is a globule formulation. In some embodiments, a provided formulation is a candy formulation. In some embodiments, a provided formulation is a capsule formulation. In some embodiments, a provided formulation may be in the form of a dry powder (e.g., a dry spray). In some embodiments, a provided formulation may be in the form of a solid particle suspension. In some embodiments, a provided formulation may be in the form of a fast-dissolving tablet. In some embodiments, a provided formulation may be in the form of a fast-dissolving film.

[0193] In various embodiments, a polypeptide formulation as described herein, e.g., a truffle formulation, a tablet formulation, a globule formulation, a candy formulation, a capsule formulation, can include a coating. An enteric coating is typically a barrier that controls the location of a recipients body (e.g., digestive system, e.g., gut, e.g., small and / or large intestine) in which an oral formulation’s core is exposed and / or to which the oral formulation’s core is delivered. Many enteric coatings are insoluble at a low pH but dissolve, swells, or becomes soluble at a higher pH in the intestinal tract. Typical materials used for enteric coatings can include CAP (cellulose acetate phthalate), CAT (cellulose acetate trimellitate), PVAP (poly(vinyl acetate phthalate)) and HPMCP (hydroxypropyl methylcellulose phthalate), poly(methacrylic acid-co-methyl methacrylate), fatty acids, waxes, shellac (e.g., esters of aleurtic acid), plastics, and plant fibers. In various embodiments, the dissolution pH of an enteric coating can be between, e.g., about 4.5 and about 7, e.g., about 4.5 to about 5.5, about 4.5 to about 6.0, about 5.5 to about 7.0, about 5.0, about 6.2, or about 7.0. In various embodiments an enteric coating has a pH release that is within a range that has a lower bound of about 4.5, 5.0, 5.5, or 6.0 and an upper bound of about 5.0, 5.5, 6.0, 6.5, 7.0, or 7.5. In some embodiments, a coating is or comprises an enteric coating. In some embodiments, a coating is or comprises Vitamin E.

[0194] The present disclosure expressly includes the recognition that enteric coatings are not necessary for delivery of a polypeptide disclosed herein according to various embodiments methods and compositions as described herein to reach the bloodstream, plasma, lymphatic system, and / or thoracic duct, e.g., in therapeutically effective amounts.

[0195] In various embodiments, a formulation that includes a crystallized polypeptide composition is or includes a bioadhesive formulation, e.g., in or on the surface of a formulation, e.g., truffle, tablet, globule, candy, or capsule formulation. In various embodiments, a bioadhesive formulation adheres to a specific biological location such as a mucosal lining (mucoadhesion). Bioadhesive dosage forms can improve the oral absorption of polypeptide agent by delivering it in small doses over an extended period and / or localizing it in the intestine by bioadhesion. Various bioadhesive polymers can be broadly as specific or nonspecific. Specific bioadhesive polymers (e.g., lectins, and fimbrins) have the ability to adhere to specific chemical structures within the biological molecules while the nonspecific bioadhesive polymers (e.g., polyacrylic acid [PAA] and cyanoacrylates) have the ability to bind with both the cell surfaces and the mucosal layer. Further examples of bioadhesive polymers include CMC sodium, Carbopol, Polycarbophil, Tragacanth, Sodium alginate, HPMC, Gum karaya, Gelatin, Guar gum, Pectin, Acacia, Chitosan, and hydroxypropyl cellulose. Examples of bioadhesive polymers can include Hydrophilic polymers (e.g., Methyl Cellulose, hydroxyethyl cellulose, HPMC, Na CMC, and carbomers), Thiolated polymers (e.g., Chitosan-iminothiolane, PAA-cysteine, PAA-homocysteine, chitosan- thioglycolic acid, chitosan-thioethylamidine, alginate-cysteine, poly (methacrylic acid)- cysteine and sodium carboxymethylcellulose-cysteine), Lectin-based polymers (e.g., Lentil lectin, peanut agglutinin, and ulex europaeus agglutinin), Polyox WSR (e.g., WSR N-10, WSR N-80, WSR N-205, and WSR N-750), and other polymers such as tomato lectin, PAA- co-PEG, and PSA.

[0196] The present disclosure includes methods and compositions that include a polypeptide formulation for oral delivery including a core within a pharmaceutically acceptable shell, where the core includes a polypeptide composition (e.g., a crystallized polypeptide composition).

[0197] In various embodiments, a core comprises about 1 mg to 2,000 mg of carrier polypeptide (e.g., crystalized polypeptide or amorphous polypeptide). In various embodiments, a core comprises about 1 mg to 1,000 mg, 1 mg to 500 mg, 1 mg to 400 mg, 1 mg to 300 mg, 1 mg to 200 mg, or 1 mg to 100 mg of crystalized polypeptide or amorphous carrier polypeptide. In some embodiments the formulation includes an amount of crystalizedpolypeptide or amorphous polypeptide that is within a range having a lower bound of, e.g., 1, 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 75, 100, 150, 200, 250, 300, 350, 400, 450, or 500 mg crystalized polypeptide or amorphous polypeptide and an upper bound of 100, 150, 200, 250, 300, 350, 400, 450, 500, 1,000, or 2,000 mg crystalized polypeptide or amorphous polypeptide. In various embodiments, a core comprises about 50 pg to 2,000 mg of carrier polypeptide, optionally wherein the polypeptide formulation includes about 50 pg to 1,000 mg, 50 pg to 500 mg, 50 pg to 400 mg, 50 pg to 300 mg, 50 pg to 200 mg, 50 pg to 100 mg, 50 pg to 50 mg, 50 pg to 25 mg, 50 pg to 20 mg, 50 pg to 15 mg, 50 pg to 10 mg, 50 pg to 5 mg, 50 pg to 1 mg, 50 pg to 500 pg, 1 mg to 1,000 mg, 1 mg to 500 mg, 1 mg to 400 mg, 1 mg to 300 mg, 1 mg to 200 mg, 1 mg to 100 mg, 1 mg, to 50 mg, 1 mg to 25 mg crystalized polypeptide or amorphous polypeptide. In various embodiments, a core comprises about 1 pg to 2,000 mg of carrier polypeptide, optionally wherein the polypeptide formulation includes about 1 pg to 1,000 mg, 1 pg to 500 mg, 1 pg to 400 mg, 1 pg to 300 mg, 1 pg to 200 mg, 1 pg to 100 mg, 1 pg to 50 mg, 1 pg to 25 mg, 1 pg to 20 mg, 1 pg to 15 mg, 1 pg to 10 mg, 1 pg to 5 mg, 1 pg to 1 mg, 1 pg to 500 pg, 1 pg to 250 pg, 1 pg to 200 pg, 1 pg to 150 pg, 1 pg to 100 pg, 1 pg to 50 pg of the crystalized polypeptide or amorphous polypeptide. In some embodiments the formulation includes an amount of crystalized carrier polypeptide or amorphous carrier polypeptide that is within a range having a lower bound of, e.g., 1 pg, 5 pg, 10 pg, 15 pg, 20 pg, 25 pg 50 pg, 100 pg, 200 pg, 300 pg, 400 pg, 500 pg, 600 pg, 700 pg, 800 pg, 900 pg, 1 mg, 2 mg, 3 mg, 4 mg, 5 mg, 10 mg, or 25 mg crystalized polypeptide or amorphous polypeptide and an upper bound of 500 pg, 600 pg, 700 pg, 800 pg, 900 pg, 1 mg, 2 mg, 3 mg, 4 mg, 5 mg, 10 mg, 25 mg, 50 mg, 100 mg, 500 mg, 1,000 mg, or 2,000 mg crystalized polypeptide or amorphous polypeptide.

[0198] In various embodiments, a core comprises about 1 mg to 200 mg of polypeptide (e.g., crystalized semaglutide polypeptide). In various embodiments, a core comprises about 1 mg to 100 mg, 1 mg to 50 mg, 1 mg to 25 mg, 1 mg to 15 mg, 1 mg to 10 mg, or 1 mg to 5 mg of crystalized polypeptide. In some embodiments the formulation includes an amount of crystalized polypeptide that is within a range having a lower bound of, e.g., 1, 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 75, 100, 150, 200, 250, 300, 350, 400, 450, or 500 mg crystalized polypeptide and an upper bound of 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 100, or 200 mg crystallized polypeptide.

[0199] In various embodiments, a core comprises about 500 pg to 200 mg of active polypeptide, optionally wherein the polypeptide formulation includes about 500 pg to 150 mg, 500 pg to 100 mg, 500 pg to 75 mg, 500 pg to 50 mg, 500 pg to 25 mg, 500 pg to 15mg, 500 pg to 10 mg, 500 pg to 5 mg, 1 mg to 150 mg, 1 mg to 100 mg, 1 mg to 75 mg, 1 mg to 50 mg, 1 mg to 25 mg, 1 mg to 15 mg, 1 mg, to 10 mg, 1 mg to 5 mg crystallized polypeptide. In various embodiments, a core comprises about 100 pg to 200 mg of active polypeptide, optionally wherein the polypeptide formulation includes about 100 pg to 150 mg, 100 pg to 100 mg, 100 pg to 75 mg, 100 pg to 50 mg, 100 pg to 25 mg, 100 pg to 15 mg, 100 pg to 10 mg, 100 pg to 5 mg, of the crystallized polypeptide. In some embodiments the formulation includes an amount of crystalized active polypeptide that is within a range having a lower bound of, e.g., 10 pg, 50 pg, 100 pg, 150 pg, 200 pg, 250 pg, 500 pg, 1 mg, 2 mg, 3 mg, 4 mg, 5 mg, 6 mg, 7 mg, 8 mg, 9 mg, 10 mg, or 25 mg crystallized polypeptide and an upper bound of 500 pg, 600 pg, 700 pg, 800 pg, 900 pg, 1 mg, 2 mg, 5 mg, 10 mg, 25 mg, 50 mg, 100 mg, or 200 mg crystallized polypeptide.

[0200] In various embodiments, one or more pharmaceutically acceptable carriers are present in a formulation (e.g., a truffle, a tablet, a globule, a candy, or a capsule formulation) or a core thereof at a concentration between 0.1 mM and about 1,000 mM, between about 0.1 mM and about 500 mM, between about 0.1 mM and about 200 mM, or between about 1 mM and about 100 mM. In various embodiments, one or more pharmaceutically acceptable carriers are present in a formulation (e.g., a truffle, a tablet, a globule, or a candy formulation) or a core thereof at a concentration within a range having a lower bound of 0.1, 1, 5, 10, 20, 30, 40, 50, 75, or 100 mM and an upper bound of 10, 20, 30, 40, 50, 75, 100, 150, 200, 250 or 500 mM.

[0201] In some embodiments, a polypeptide formulation as described herein is in a shape of sphere, cube, cone, cylinder, half sphere, torus, pyramid, triangular prism, hexagonal prism, cuboid, hexagonal pyramid, hallow cylinder, octahedron, diamond, star prism, hexagonal diamond, star pyramid, pentagonal prism, L shape prism, dodecahedron, tetrahedron, or icosahedron, or a modification thereof, or a combination thereof.

[0202] Those skilled in the art, reading the present disclosure will appreciate that, in some embodiments, a suitable means of administration for a particular provided formulation to a particular subject may be selected upon consideration, among other things, of, for example age and / or condition of a subject. Similarly, in some embodiments, unit dose, appropriate dosing regimen, and / or total dose to be administered may be selected based on sound medical judgment taking into consideration, for example, any designated or approved range is provided, weight, age, condition, and other characteristics of a patient, etc.

[0203] Those skilled in the art, reading the present disclosure will appreciate that, in some embodiments, it may be feasible to administer certain provided formulations by otherroutes. In some embodiments, certain provided formulations (e.g., truffle, tablet, globule, candy formulations) can be administered rectally as suppository formulations.EXEMPLARY FORMULATIONSTruffle Formulations

[0204] In some embodiments, the present disclosure provides truffle formulations comprising a core and a pharmaceutically acceptable shell.

[0205] In some embodiments, a core comprises (1) a semaglutide polypeptide composition (e.g., a crystalline semaglutide polypeptide composition which may, in some embodiments, be a powder); and (2) one or more pharmaceutically acceptable carriers or excipients. In some embodiments, provided oral formulations include (3) a carrier polypeptide composition (e.g., a crystalline carrier polypeptide composition). In a provided truffle formulation, such a core can be situated in a shell. In some embodiments, a core comprises crystallized carrier polypeptide composition. In some embodiments, a core comprises crystallized active polypeptide composition. In some embodiments, a core comprises an amorphous carrier polypeptide composition.

[0206] In some embodiments, a shell is or comprises sugar. In some embodiments, a shell is or comprises cane sugar. In some embodiments, a shell is or comprises palm sugar. In certain embodiments, a shell is or comprises lactose. In certain embodiments, a shell is or comprises xylitol. In certain embodiments, a shell is or comprises milk sugar. In some embodiments, a shell is cane sugar. In some embodiments, a shell is palm sugar.

[0207] In some embodiments, a shell includes hardshell and soft shell. In various embodiments, a shell is a gelatin shell or hydroxypropyl methylcellulose (HPMC) shell. Exemplary shell materials can include polymers such as poly(glycolic acid) (PGA), poly(lactic acid) (PLA) and its copolymers, poly(lactide-co-glycolide) (PLGA), and nonionic cellulose ethers such as hydroxypropylcellulose (HPC) and hydroxypropyl methylcellulose (HPMC). In some embodiments a capsule is a vegetable capsule.

[0208] In some embodiments, shell of a truffle formulation can be produced from a single piece of gelatin. Gelatin can be used to surround a core not based on water, as water would dissolve the gelatin. Once ingested, the shell dissolves, exposing the core.

[0209] In some embodiments, shell of a truffle formulation can be filled with a core including dry ingredients in powder form. In some such embodiments, a shell may be formed, its body is first filled with a core composition, and then the shell closed with the cap. Once ingested, the shell dissolves, exposing the core.

[0210] In some embodiments, a shell is hollow. In some embodiments, a shell is partially filled with space of a core. In some embodiments, a shell is partially fully with space of a core.

[0211] In some embodiments, a polypeptide formulation as described herein is in a shape of sphere, or a modification thereof. In some embodiments, a truffle formulation as described herein is in a shape of sphere, or a modification thereof.Tablet formulations

[0212] In some embodiments, the present disclosure provides formulations comprising (1) a semaglutide polypeptide composition (e.g., a crystalline semaglutide polypeptide composition which may, in some embodiments, be a powder); and (2) one or more pharmaceutically acceptable carriers or excipients. In some embodiments, provided formulations include (3) a carrier polypeptide composition (e.g., a crystalline carrier polypeptide composition). In some embodiments, a tablet formulation as described herein is prepared by pressing a mixture of (1) a semaglutide polypeptide composition (e.g., a crystalline semaglutide polypeptide composition which may, in some embodiments, be a powder); and (2) one or more pharmaceutically acceptable carriers or excipients, and optionally (3) a carrier polypeptide composition (e.g., a crystalline carrier polypeptide composition).

[0213] In some embodiments, the present disclosure provides formulations that include one or more of crystalline semaglutide, microcrystalline cellulose, providone K90, Vitamin E, magnesium stearate, dextrate, and crospovidon, or any combinations thereof.

[0214] In some embodiments, the present disclosure provides formulations that include crystalline semaglutide, microcrystalline cellulose, providone K90, Vitamin E, and magnesium stearate.

[0215] In some embodiments, the present disclosure provides formulations that include crystalline semaglutide, microcrystalline cellulose, providone K90, and magnesium stearate.

[0216] In some embodiments, the present disclosure provides formulations that include crystalline semaglutide, dextrate, and crospovidon.

[0217] In some embodiments, the present disclosure provides formulations that include crystalline semaglutide, microcrystalline cellulose, and Vitamin E.

[0218] In some embodiments, the present disclosure provides formulations that include about .5%, 1%, 1.5%, 2%, 2.5%, 3%, 4%, or 4.5%, weight per weight (w / w) crystalline semaglutide.

[0219] In some embodiments, the present disclosure provides formulations that include at least .5%, 1%, 1.5%, 2%, weight per weight (w / w) crystalline semaglutide.

[0220] In some embodiments, the present disclosure provides formulations that include about 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, or 25% weight per weight (w / w) crystalline semaglutide.

[0221] In some embodiments, the present disclosure provides formulations that include at least 8%, 9%, 10%, 11%, 12%, or 13% weight per weight (w / w) crystalline semaglutide.

[0222] In some embodiments, the present disclosure provides formulations that include between .5% and 4% weight per weight (w / w) crystalline semaglutide.

[0223] In some embodiments, the present disclosure provides formulations that include between 8% and 15% weight per weight (w / w) crystalline semaglutide.

[0224] In some embodiments, the present disclosure provides formulations that include between .5% and 25% weight per weight (w / w) crystalline semaglutide.

[0225] In some embodiments, the present disclosure provides formulations that include about 2.0% w / w crystalline semaglutide.

[0226] In some embodiments, the present disclosure provides formulations that include about 13.0% w / w crystalline semaglutide.

[0227] In some embodiments, the present disclosure provides formulations that include about 1%, 1.5%, 2%, 2.5%, 3%, 3.5%, 4%, 4.5%, 5%, 5.5%, or 6% weight per weight (w / w) Vitamin E agent.

[0228] In some embodiments, the present disclosure provides formulations that include at least 1%, 1.5%, 2%, 2.5%, 3%, or 3.5% weight per weight (w / w) Vitamin E agent.

[0229] In some embodiments, the present disclosure provides formulations that include between 1% and 6% w / w Vitamin E agent.

[0230] In some embodiments, the present disclosure provides formulations that include about 3.7% w / w Vitamin E agent.

[0231] In some embodiments, the present disclosure provides formulations that include about 60%, 65%, 70%, 75%, 80%, 85%, or 90% weight per weight (w / w) microcrystalline cellulose.

[0232] In some embodiments, the present disclosure provides formulations that include at least 60%, 65%, 70%, 75%, 80%, or 85% weight per weight (w / w) microcrystalline cellulose.

[0233] In some embodiments, the present disclosure provides formulations that include between 60% and 95% w / w microcrystalline cellulose.

[0234] In some embodiments, the present disclosure provides formulations that include about 89% w / w microcrystalline cellulose.

[0235] In some embodiments, the present disclosure provides formulations that include about 93% w / w microcrystalline cellulose.

[0236] In some embodiments, the present disclosure provides formulations that include about .5%, 1%, 1.5%, 2%, 2.5%, 3%, 3.5%, 4%, 4.5%, or 5% weight per weight (w / w) providone K90.

[0237] In some embodiments, the present disclosure provides formulations that include at least .5%, 1%, 1.5%, or 2% weight per weight (w / w) providone K90.

[0238] In some embodiments, the present disclosure provides formulations that include between .5% and 5% w / w providone K90.

[0239] In some embodiments, the present disclosure provides formulations that include about 2% w / w providone K90 .

[0240] In some embodiments, the present disclosure provides formulations that include about .5%, 1%, 1.5%, 2%, 2.5%, 3%, 3.5%, 4%, 4.5%, 5%, 5.5%, or 6% weight per weight (w / w) magnesium stearate.

[0241] In some embodiments, the present disclosure provides formulations that include at least .5%, 1%, 1.5%, or 2% weight per weight (w / w) magnesium stearate.

[0242] In some embodiments, the present disclosure provides formulations that include between .5% and 6% w / w magnesium stearate.

[0243] In some embodiments, the present disclosure provides formulations that include about 2.5% w / w magnesium stearate.

[0244] In some embodiments, the present disclosure provides formulations that include about 60%, 65%, 70%, 75%, 80%, 85%, 90%, or 95% weight per weight (w / w) dextrose.

[0245] In some embodiments, the present disclosure provides formulations that include at least 60%, 65%, 70%, or 75%, weight per weight (w / w) dextrate.

[0246] In some embodiments, the present disclosure provides formulations that include between 60% and 95% w / w dextrate.

[0247] In some embodiments, the present disclosure provides formulations that include about 80% w / w dextrate.

[0248] In some embodiments, the present disclosure provides formulations that include about 3%, 4%, 4.5%, 5%, 5.5%, 6%, 6.5%, 7%, 7.5%, 8%, 9%, 9.5%, 10%, 10.5%, 11%, 11.5%, 12%, 12.5%, or 13% weight per weight (w / w) crospovidon.

[0249] In some embodiments, the present disclosure provides formulations that include at least 3%, 4%, 4.5%, 5%, 5.5%, 6%, 6.5%, 7% weight per weight (w / w) crospovidon.

[0250] In some embodiments, the present disclosure provides formulations that include between 3% and 13% w / w crospovidon.

[0251] In some embodiments, the present disclosure provides formulations that include about 7.3% w / w crospovidon.

[0252] .Alternatively or additionally, in some embodiments, a tablet formulation as described herein is coated with a pharmaceutically acceptable coating. In some embodiments, a coating is or comprises food ingredient. In some embodiments, a coating is or comprises an enteric coating. In some embodiments, a coating is or comprises Vitamin E.Globule formulations

[0253] In some embodiments, the present disclosure provides a globule formulation comprising (1) a semaglutide polypeptide composition (e.g., a crystalline semaglutide polypeptide composition which may, in some embodiments, be a powder); and (2) one or more pharmaceutically acceptable carriers or excipients. In some embodiments, provided oral formulations include (3) a carrier polypeptide composition (e.g., a crystalline carrier polypeptide composition). In some embodiments, a globule formulation comprises asuspension of solid particles, wherein such solid particles comprise a crystallized or an amorphous carrier and active polypeptide compositions and optionally one or more pharmaceutically acceptable carriers.

[0254] Alternatively or additionally, in some embodiments, a globule formulation as described herein is coated with a pharmaceutically acceptable coating. In some embodiments, a coating is or comprises food ingredient. In some embodiments, a coating is or comprises an enteric coating.Candy formulations

[0255] In some embodiments, the present disclosure provides a candy formulation comprising (1) a semaglutide polypeptide composition (e.g., a crystalline semaglutide polypeptide composition which may, in some embodiments, be a powder); and (2) one or more pharmaceutically acceptable carriers or excipients. In some embodiments, provided oral formulations include (3) a carrier polypeptide composition (e.g., a crystalline carrier polypeptide composition).

[0256] In some embodiments, a candy formulation comprises one or more food ingredients, e.g., chocolate, cocoa, milk, milk product, natural dye, artificial dye, gum base, flavors, sweeteners, gelatin, starch, syrup, citric acid, sugar, etc.

[0257] In some embodiments, a candy formulation comprises about 50% w / w of sugar. In some embodiments, a candy formulation comprises about 60% w / w of sugar. In some embodiments, a candy formulation comprises about 70% w / w of sugar. In some embodiments, a candy formulation comprises about 80% w / w of sugar. In some embodiments, a candy formulation comprises about 90% w / w of sugar. In some embodiments, a candy formulation comprises about 91% w / w of sugar. In some embodiments, a candy formulation comprises about 92% w / w of sugar. In some embodiments, a candy formulation comprises about 93% w / w of sugar. In some embodiments, a candy formulation comprises about 94% w / w of sugar. In some embodiments, a candy formulation comprises about 95% w / w of sugar. In some embodiments, a candy formulation comprises about 96% w / w of sugar. In some embodiments, a candy formulation comprises about 97% w / w of sugar. In some embodiments, a candy formulation comprises about 98% w / w of sugar. In some embodiments, a candy formulation comprises about 99% w / w of sugar.

[0258] In some embodiments, a candy formulation comprises more than about 50% w / w of sugar. In some embodiments, a candy formulation comprises more than about 60w / w of sugar. In some embodiments, a candy formulation comprises more than about 70% w / w of sugar. In some embodiments, a candy formulation comprises more than about 80% w / w of sugar. In some embodiments, a candy formulation comprises more than about 90% w / w of sugar. In some embodiments, a candy formulation comprises more than about 95% w / w of sugar. In some embodiments, a candy formulation comprises more than about 99% w / w of sugar. In some embodiments, a candy formulation comprises more than about 99.9% w / w of sugar.

[0259] In some embodiments, a candy formulation comprises less than about 50% w / w of sugar. In some embodiments, a candy formulation comprises less than about 40 w / w of sugar. In some embodiments, a candy formulation comprises less than about 30% w / w of sugar. In some embodiments, a candy formulation comprises less than about 20% w / w of sugar. In some embodiments, a candy formulation comprises less than about 10% w / w of sugar. In some embodiments, a candy formulation comprises less than about 5% w / w of sugar. In some embodiments, a candy formulation comprises less than about 3% w / w of sugar. In some embodiments, a candy formulation comprises less than about 2% w / w of sugar. In some embodiments, a candy formulation comprises less than about 1% w / w of sugar. In some embodiments, a candy formulation comprises less than about 0.5% w / w of sugar. In some embodiments, a candy formulation comprises less than about 0.1% w / w of sugar.

[0260] In some embodiments, a candy formulation is in a form of, for example, of candy gems, chewing gum, gummy candy, hard candy (e.g., drops, lollipops, lozenges, rock candy, stick candy, etc.) marshmallows, syrup, toffee, etc.. In some embodiments, a candy formulation may be in the form of a drop, film, gel, patch, spray, wafer, etc.Capsule formulations

[0261] In some embodiments, the present disclosure provides a capsule formulation comprising (1) a semaglutide polypeptide composition (e.g., a crystalline semaglutide polypeptide composition which may, in some embodiments, be a powder); and (2) one or more pharmaceutically acceptable carriers or excipients , optionally wherein a pharmaceutically acceptable carrier comprises a Vitamin E agent. In some embodiments, provided oral formulations include (3) a carrier polypeptide composition (e.g., a crystalline carrier polypeptide composition).

[0262] In some embodiments, a core including (1) a semaglutide polypeptide composition (e.g., a crystalline semaglutide polypeptide composition which may, in someembodiments, be a powder); and (2) one or more pharmaceutically acceptable carriers or excipients can be encapsulated in a polymer capsule. In some embodiments, a capsule formulation includes pharmaceutically acceptable capsules suitable for oral administration or rectal administration. In some embodiments, provided capsule formulations are capsules suitable for delivery, e.g., specific delivery, of a polypeptide to the gut. In various embodiments, capsules of the present disclosure are capsules suitable for delivery, e.g., specific delivery, of a polypeptide to one or both the small intestine and / or large intestine.

[0263] Capsules of the present disclosure include hard-shell capsules and soft-shell capsules. In various embodiments, a capsule is a gelatin capsule or hydroxypropyl methylcellulose (HPMC) capsule. Exemplary capsule materials can include polymers such as poly(glycolic acid) (PGA), poly(lactic acid) (PLA) and its copolymers, poly(lactide-co- glycolide) (PLGA), and non-ionic cellulose ethers such as hydroxypropylcellulose (HPC) and hydroxypropyl methylcellulose (HPMC). In some embodiments a capsule is a vegetable capsule.

[0264] Soft gelatin capsules, also called softgels, can be produced from a single piece of gelatin. They can be used to encapsulate solutions not based on water, as water would dissolve the gelatin. Once ingested, the capsule dissolves, exposing the core.

[0265] Hard gelatin capsules are made of two parts, a body and a cap. Hard gelatin capsules can be filled with a core including dry ingredients in powder form. The body is first filled with a core composition, and the capsule is then closed with the cap. Once ingested, the hard capsules dissolves, exposing the core.Suppository formulations

[0266] The present disclosure further includes suppositories suitable for rectal delivery. In various embodiments, suppositories melt or dissolve upon administration. In various embodiments, a suppository includes a shell (e.g., a cane sugar shell or a capsule shell disclosed herein) that surrounds a core. In various embodiments, a suppository does not include a shell-core structure. Suppositories can be formed from waxy matter, structured glycerine, hydrogenated vegetable oil, polyethylene glycol wax derivative, or poloxamer- based mixtures. In various embodiments, a suppository is solid at ambient temperature but rapidly melts at body temperature. Emulsifiers may be used to increase the solubility of crystallized polypeptide or amorphous polypeptide in the suppository mass and / or accelerate the dispersal of crystallized polypeptide or amorphous polypeptide after the suppository melts.

[0267] Certain suppository formulations can include a crystallized polypeptide composition and suppository excipients, e.g., a lipophilic base (e.g., cocoa butter, coconut oil, virgin coconut oil, almond oil, wheatgerm oil, any edible oil, hydrogenated vegetable oils, and hard fats) or hydrophilic base (e.g., glycerinated gelatin and polyethylene glycols). Lipophilic bases are immiscible with body fluids and readily melt at body temperature to release the drug on the mucosal surface, whereas hydrophilic bases need to dissolve in the physiological fluids for drug release. Suppository formulations can include solid, semi-solid, and liquid forms.

[0268] In various embodiments, a suppository formulation includes a semi-solid dosage form such as a gel or foam. A rectal gel can be a semi-solid formulations that contain a solvent trapped within a polymer network to create a viscous consistency. Viscosity of a gel can be modified by the addition of co-solvents (e.g., glycerin and propylene glycol) and electrolytes.

[0269] In various embodiments, a suppository formulation includes a liquid suppository, e.g., a liquid suppository including thermosensitive polymers (e.g., poloxamers), mucoadhesive polymers (e.g., carbopol, sodium alginate, polycarbophil, hydroxypropyl methylcellulose, hydroxyethyl cellulose, and methylcellulose), or a combination of thermosensitive and mucoadhesive polymers. Suppositories can further include, e.g., cellulose ether polymers (e.g., hydroxypropyl methylcellulose, hydroxyethyl cellulose, and methy Icellulo se) .

[0270] In various embodiments, a suppository formulation includes a foam such as a colloidal dosage form with a hydrophilic liquid continuous phase containing a foaming agent and a gaseous dispersion phase distributed throughout. Following rectal administration, certain such formulations transition from a foam state to a liquid or semi- solid state on the mucosal surface. Foaming agents include amphiphilic substances that are important for foam generation and stabilization.

[0271] Polypeptide formulations as described herein are suitable for oral administration or rectal administration.Table 2: Exemplary semaglutide sequencesPOLYPEPTIDE DELIVERY AND APPLICATIONS

[0272] Compositions and methods provided by present disclosure include formulations for oral administration to a subject, which oral administration, in various embodiments, results in delivery of a polypeptide to e.g., thoracic duct, lymphatic system, small and / or large intestine, and / or bloodstream of a subject.

[0273] Without wishing to be bound by any particular scientific theory, the present inventors have surprisingly discovered that polypeptides orally administered polypeptide formulations as described herein, e.g., truffle, tablet, globule, candy, or capsule formulations, are efficiently delivered to the bloodstream, e.g., via buccal / sublingual administration.Accordingly, the present disclosure provides a platform or system of general, including compositions and methods disclosed herein, for delivery of diverse polypeptides to the thoracic duct, lymphatic system, small and / or large intestine, and / or blood stream.

[0274] The present disclosure provides compositions and methods, e.g., for delivery of a polypeptide to a subject, e.g., to the thoracic duct, lymphatic system, small and / or largeintestine, and / or blood stream of the subject. In various embodiments, composition or method as described herein delivers polypeptide to bloodstream with beneficial pharmacokinetic characteristics. In various embodiments, beneficial pharmacokinetic characteristics can result from distribution of a polypeptide through mouth tissue, e.g., via buccal / sublingual administration. Accordingly, the present disclosure specifically includes methods and compositions for delivery of a polypeptide to mouth tissue, e.g., via buccal / sublingual administration.

[0275] In various embodiments, a polypeptide of a provided formulation is characterized in that after administration according to the present disclosure, a polypeptide delivered to bloodstream thereby has a median, mean, or modal half-life in a plurality of serum samples, systems, and / or across polypeptide molecules after administration (e.g., across a plurality of subjects after oral administration of a polypeptide formulation to each subject) of at least 0.5 hours, optionally wherein the half-life is at least 1, 2, 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60 hours, or more, optionally wherein the median, mean, or modal half-life is in a range having a lower bound of 1, 2, 5,10, or 15 hours and an upper bound of 20, 25, 30, 35, 40, 45, 50, 55, 60 hours, or more. In various embodiments, after oral administration to a subject of a polypeptide formulation as described herein, the active polypeptide delivered to bloodstream by a provided formulation has a half-life of at least 0.5 hours, optionally wherein the half-life is at least 1, 2, 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, or 60 hours, or more, optionally wherein the half-life is in a range having a lower bound of 1, 2, 5, 10, or 15 hours and an upper bound of 20, 25, 30, 35, 40, 45, 50, 55, 60 hours, or more.

[0276] In various embodiments, a polypeptide of a provided formulation is characterized in that after administration according to the present disclosure, a polypeptide delivered to bloodstream by such provided formulation has a median, mean, or modal halflife across a plurality of samples, systems, and / or polypeptide molecules (e.g., across a plurality of subjects after oral administration of the polypeptide formulation to each subject) that is at least 10% greater than the median, mean, or modal half-life achieved by injection (e.g., intravenous administration) of the polypeptide (e.g., according to a standard of care route of administration for the polypeptide), optionally wherein the median, mean, or modal half-life of the polypeptide delivered to bloodstream by a provided formulation (e.g., after oral administration) is at least 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 2-fold, 3- fold, or 4-fold, greater than the median, mean, or modal half-life achieved by injection (e.g., intravenous administration) of the polypeptide (e.g., according to a standard of care route of administration for the polypeptide). In various embodiments, after oral administration to asubject of the polypeptide formulation, the active polypeptide delivered to bloodstream by a provided formulation has a half-life that is at least 10% greater than the half-life achieved by injection (e.g., intravenous administration) of the polypeptide (e.g., according to a standard of care route of administration for the polypeptide), optionally wherein the half-life of the polypeptide delivered to bloodstream by a provided formulation after oral administration to the subject is at least 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 2-fold, 3-fold, or 4-fold, greater than the half-life achieved by injection (e.g., intravenous administration) of the polypeptide (e.g., according to a standard of care route of administration for the polypeptide).

[0277] In various embodiments, a polypeptide of a provided formulation is characterized in that after administration according to the present disclosure, a polypeptide delivered thereby has a median, mean, or modal Tmaxacross a plurality of samples, systems, and / or polypeptide molecules (e.g., across a plurality of subjects after oral administration of the polypeptide formulation to each subject) that is at least 10% greater than the median, mean, or modal Tmaxachieved by injection (e.g., intravenous administration) of the polypeptide (e.g., according to a standard of care route of administration for the polypeptide), optionally wherein the median, mean, or modal Tmaxof the polypeptide of the polypeptide formulation (e.g., after oral administration) is at least 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 2-fold, 3 -fold, or 4-fold, 5-fold, 10-fold, or 20-fold greater than the median, mean, or modal Tmaxachieved by injection (e.g., intravenous administration) of the polypeptide (e.g., according to a standard of care route of administration for the polypeptide). In various embodiments, after oral administration to a subject of the polypeptide formulation, the active polypeptide of the polypeptide formulation has a Tmaxthat is at least 10% greater than the Tmaxachieved by injection (e.g., intravenous administration) of the polypeptide (e.g., according to a standard of care route of administration for the polypeptide), optionally wherein the Tmaxof the polypeptide of the polypeptide formulation after oral administration to the subject is at least 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 2-fold, 3-fold, or 4-fold, 5-fold, 10-fold, or 20-fold greater than the Tmaxachieved by injection (e.g., intravenous administration) of the polypeptide (e.g., according to a standard of care route of administration for the polypeptide).

[0278] In various embodiments, activea polypeptide formulation is characterized in that after administration according to the present disclosure, an activepolypeptide delivered thereby has a median, mean, or modal bioavailability across a plurality of samples, systems, and / or polypeptide molecules (e.g., across a plurality of subjects after oral administration of the polypeptide formulation to each subject) that is at least 1% ofthe median, mean, or modalbioavailability achieved by injection (e.g., subcutaneous administration) of the polypeptide (e.g., according to a standard of care route of administration for the polypeptide), optionally wherein the median, mean, or modal bioavailability of the polypeptide of the polypeptide formulation (e.g., after oral administration) is at least 1%, 1.5%, 2%, 3%, 4%, 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 2-fold, 3-fold, 4-fold, or 5-fold of the median, mean, or modal bioavailability achieved by injection (e.g., subcutaneous administration) of the polypeptide (e.g., according to a standard of care route of administration for the polypeptide). In various embodiments, after oral administration to a subject of a polypeptide formulation, the active polypeptide of the polypeptide formulation has an bioavailability that is at least 1% of the bioavailability achieved by injection (e.g., subcutaneous administration) of the polypeptide (e.g., according to a standard of care route of administration for the polypeptide) , optionally wherein the bioavailability of the polypeptide of the polypeptide formulation after oral administration to the subject is at least 1%, 1.5%, 2%, 3%, 4%, 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 2-fold, 3-fold, 4-fold, or 5-fold of the bioavailability achieved by injection (e.g., subcutaneous administration) of the polypeptide (e.g., according to a standard of care route of administration for the polypeptide). In various embodiments, a polypeptide of a polypeptide formulation is characterized in that after administration according to the present disclosure, a polypeptide delivered thereby has a median, mean, or modal bioavailability across a plurality of samples, systems, and / or polypeptide molecules (e.g., across a plurality of subjects after oral administration of the polypeptide formulation to each subject) that is at least 1%, e.g., at least 1%, 1.5%, 2%, 3%, 4%, 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%. In some embodiments, provided formulations achieve oral bioavailability above about 5%, or within a range of about 10%-20% oral bioavailability, or about 15%.

[0279] In various embodiments, a polypeptide of a polypeptide formulation is characterized in that after administration according to the present disclosure, a polypeptide delivered thereby has a median, mean, or modal Cmax across a plurality of samples, systems, and / or polypeptide molecules (e.g., across a plurality of subjects after oral administration of the polypeptide formulation to each subject) that is at least 10% less than the median, mean, or modal Cmax achieved by injection (e.g., subcutaneous administration) of the polypeptide (e.g., according to a standard of care route of administration for the polypeptide), optionally wherein the median, mean, or modal Cmax of the polypeptide of the polypeptide formulation (e.g., after oral administration) is at least 20%, 30%, 40%, 50%, 60%, 70%, 80%, or 90%, less than the median, mean, or modal Cmax achieved by injection (e.g., subcutaneousadministration) of the polypeptide (e.g., according to a standard of care route of administration for the polypeptide). In various embodiments, after oral administration to a subject of a polypeptide formulation, the active polypeptide of the polypeptide formulation has a Cmax that is at least 10% less than the Cmax achieved by injection (e.g., subcutaneous administration) of the polypeptide (e.g., according to a standard of care route of administration for the polypeptide), optionally wherein the Cmax of the active polypeptide of the polypeptide formulation after oral administration to the subject is at least 20%, 30%, 40%, 50%, 60%, 70%, 80%, or 90%, less than the Cmax achieved by injection (e.g., subcutaneous administration) of the polypeptide (e.g., according to a standard of care route of administration for the polypeptide).

[0280] In various embodiments, a method can include administering a composition and / or formulation as described herein to a subject in need thereof, where the composition and / or formulation includes a polypeptide for treatment of a disease or condition from which the subject suffers.

[0281] Further disclosed herein are methods of treating a disease or disorder in mammals, including administering to the mammal a composition and / or formulation as described herein that includes a therapeutically effective amount of a polypeptide, and wherein a composition and / or formulation further includes a pharmaceutically acceptable viscosity-reducing agent, aggregation-reducing agent, or other additive as described above; and wherein a therapeutic formulation is effective for treatment of a diseases or disorder. In some embodiments, an excipient compound is essentially pure.

[0282] Those skilled in the art will be aware of appropriate applications of any particular provided formulation, for example based on the polypeptide(s) it delivers and / or its therapeutic usage.

[0283] In some embodiments, a therapeutic use for a composition and / or formulation as described herein can include treatment for chronic weight management, or for improving glycemic control in adults with type 2 diabetes mellitus, and combinations thereof.

[0284] In various embodiments, the present disclosure provides a method of producing a formulation as described herein, including, e.g., steps of (1) preparing a core including a carrier polypeptide composition (e.g., an amorphous or a crystallized polypeptide composition) and a polypeptide composition (e.g., a crystallized semaglutide polypeptide composition), and (2) placing the core in a pharmaceutically acceptable shell. In various embodiments, the present disclosure provides a method of producing a tablet formulation for oral delivery, the method including mixing (i) a carrier polypeptide composition (e.g., anamorphous or a crystallized polypeptide composition) and a polypeptide composition (e.g., a crystallized semaglutide polypeptide composition) with (ii) a pharmaceutically acceptable carrier. In various embodiments, the present disclosure provides a method of producing a capsule formulation for oral delivery, the method including mixing (i) a carrier polypeptide composition (e.g., an amorphous or a crystallized polypeptide composition) and a polypeptide composition (e.g., a crystallized semaglutide polypeptide composition) with (ii) a pharmaceutically acceptable carrier. In various embodiments, the present disclosure provides a method of producing a formulation as described herein, including, e.g., steps of (1) cocrystallizing a carrier polypeptide and a polypeptide to produce a crystallized polypeptide composition; (2) preparing a core including a composition comprising a carrier polypeptide composition and a polypeptide composition (e.g., a co-crystal), and (3) placing the core in a pharmaceutically acceptable shell.EXAMPLES

[0285] The following Examples demonstrate methods and compositions for oral administration of active polypeptides that include a crystalline semaglutide polypeptide composition and a pharmaceutically acceptable carrier, which pharmaceutically acceptable carrier may be or comprise a carrier polypeptide as described herin. The following Examples further demonstrate advantages of oral formulations as described herein including improved fast dissolving tablets, half-life and bioavailability (e.g., faster onset of action), even achieving oral bioavailability that is at least about 1% or more (e.g., at least about 5%, or even 10% or more) of that observed when the semaglutide polypeptide is administered using a commereically available drug (e.g., Rybelsus®).

[0286] Furthermore, present Examples document a particularly surprising achievement of provided technologies: useful oral bioavailability of semaglutide polypeptide formulations. As is well established in the art, a variety of challenges have plagued efforts to develop orally bioavailable polypeptide formulations.

[0287] The present Examples utilize batch crystallization to produce crystallized semaglutide polypeptide compositions. As those of skill in the art will appreciate from the present disclosure that the specific production method, size, size distribution, shape, and shape distribution of crystals is not an essential feature for the successful formulation of polypeptides in accordance with methods and compositions provided herein. The presentExamples expressly demonstrate that oral formulations as described herein can achieve advantageous outcomes.Example 1: Production of Cane Sugar / Palm Sugar Truffle Shells

[0288] In order to deliver a semaglutide polypeptide through buccal or sublingual route to systemic circulation, the peptide / protein were enclosed in sugar truffles or globules. The sugar truffles or globules, which are commercially available, are made up of 100% cane sugar. Sugar truffles or globules, grade 60 (SBL Quality Globules, India), which are pharmaceutical grade, were taken and drilled to make a hole in order to produce truffle shells (Figure 1). A known amount of semaglutide polypeptide would be weighed and transferred to these shells and then sealed using palm sugar in order to create a perfect sugar truffle or globule. The semagltuide polypeptide would be inside the shell covered with sugar at this point. The shells able to hold up to 5 mg of pro tein / pep tide.

[0289] Truffles or globules work as a vehicle to transfer a semaglutide polypeptide to systemic circulation. Sugar truffles or globules do not alter the properties of drug substances. Due to their readily soluble nature, they are easy to administer in any age group. Commercial truffles or globules are available in different sizes. Their sizes are denoted in No., which start with No. 10, 20, 30, 40, etc. The diameter of size no. 10 measures 1 mm, of no. 20 measures 2 mm, and so on. So, the size of the sugar truffle shell can be altered in order to fit different amounts of semaglutide polypeptide.

[0290] Alternatively, instead of cane sugar, either lactose or xylitol or milk sugar can be used in the preparation of sugar globules / truffle shells. Cocoa truffle shells - such as 100% dark chocolate or Callebaut®’s caramelly and creamy milk chocolate, Callebaut’s creamy white chocolate or any other truffle shells can also be used to prepare the peptide / protein containing shells or tablets.Example 2: Crystallization of Semaglutide

[0291] The present Example demonstrates formation of crystallized semaglutide for use in an oral formulation as disclosed herein. Semaglutide, sold under the brand names Wegovy® and Ozempic® among others, is an antidiabetic medication used for the treatment of type 2 diabetes and long-term weight management. Semaglutide is a GLP-1 receptor agonist, meaning that it mimics the action of the human incretin glucagon-like peptide- 1 (GLP-1), thereby increasing insulin secretion and increasing blood sugar disposal andimproving glycemic control. Semaglutide is indicated as an adjunct to diet and exercise to improve glycemic control in adults with type 2 diabetes. Semaglutide is also indicated as an adjunct to diet and exercise for long-term weight management in adults with obesity. The subcutaneous injection is administered once weekly and the tablet is administered once a day. Semaglutide offers a competitive advantage over other drugs used to manage diabetes, which may require several daily doses.

[0292] Semaglutide is supplied as one ml of solution which contains 1.34 mg of semaglutide. One pre-filled pen contains 2 mg Semaglutide in 1.5 ml solution. Each dose contains 0.25 mg of Semaglutide in 0.19 ml solution. Oral Semaglutide is a tablet formulation for once-daily administration. It is co-formulated with an absorption enhancer, sodium N- (8- [2-hydroxybenzoyl] amino) caprylate (SNAC). The present Example includes the recognition that Semaglutide is exemplary of peptide / proteins for which an oral formulation would be advantageous.

[0293] The present Example utilizes semaglutide. To produce semaglutide microparticles, semaglutide solution was dialyzed against water for 24 hr with three changes and lyophilized to get dry powder. The lyophilized powder of semaglutide was then dissolved in water at a concentration of 80 mg / mL. Semaglutide aliquots were then crystalized using a variety of methods described below.

[0294] Crystallization Method 1 : A 500 pl aliquot of semaglutide (80 mg / ml), in water was mixed with 1000 pl of reagent containing 50 mM Magnesium chloride, 100 mM Tis / Hydrochloric acid pH 8.5, 40% v / v reagent alcohol and incubated at room temperature overnight. The final concentration of the semaglutide in solution was 26.67 mg / ml. This mixture was then mixed using a votex and left at room temperature. Semaglutide Micro Particles were obtained on the following day. About 80% of the input semaglutide was formed microparticles by this method.

[0295] Crystallization Method 2: A 500 pl aliquot of semaglutide (80 mg / ml), in water was mixed with 1000 pl of reagent containing 200 mM Magnesium chloride, 100 mM sodium cacodylate / Hydrochloric acid pH 6.5, 10% w / v PEG 3000 and incubated at room temperature overnight. The final concentration of the semaglutide in solution was 26.67 mg / ml. This mixture was then mixed using a votex and left at room temperature. Semaglutide Micro Particles were obtained on the following day. About 70% of the input semaglutide was formed microparticles by this method.

[0296] Crystallization Method 3: A 500 pl aliquot of semaglutide (80 mg / ml), in water was mixed with 1000 pl of reagent containing 200 mM Magnesium chloride, 100 mM Tris / Hydrochloric acid pH 7.0, 10% w / v PEG 8000 and incubated at room temperature overnight. The final concentration of the semaglutide in solution was 26.67 mg / ml. This mixture was then mixed using a votex and left at room temperature. Semaglutide Micro Particles were obtained on the following day. About 70% of the input semaglutide was formed microparticles by this method.Crystallization

[0297] Crystallization Method 4: A 500 pl aliquot of semaglutide (80 mg / ml), in water was mixed with 1000 pl of reagent containing 200 mM Magnesium chloride, 100 mM sodium cacodylate / Hydrochloric acid pH 6.5, 20% w / v PEG 1000 and incubated at room temperature overnight. The final concentration of the semaglutide in solution was 26.67 mg / ml. This mixture was then mixed using a votex and left at room temperature. Semaglutide Micro Particles were obtained on the following day. About 70% of the input semaglutide was formed microparticles by this method.

[0298] Crystallization Method 5: A 500 pl aliquot of semaglutide (80 mg / ml), in water was mixed with 1000 pl of reagent containing 200 mM Sodium chloride, 100 mM sodium phosphate dibasic / potassium phosphate monobasic pH 6.2, 40% w / v PEG 400 and incubated at room temperature overnight. The final concentration of the semaglutide in solution was 26.67 mg / ml. This mixture was then mixed using a votex and left at room temperature. Semaglutide Micro Particles were obtained on the following day. About 90% of the input semaglutide was formed microparticles by this method.

[0299] Crystallization Method 6: A 500 pl aliquot of semaglutide (80 mg / ml), in water was mixed with 1000 pl of reagent containing 200 mM Sodium chloride, 100 mM sodium phosphate dibasic / potassium phosphate monobasic pH 6.2, 35% v / v ethoxyethanol and incubated at room temperature overnight. The final concentration of the semaglutide in solution was 26.67 mg / ml. This mixture was then mixed using a votex and left at room temperature. Semaglutide Micro Particles were obtained on the following day. About 85% of the input semaglutide was formed microparticles by this method.

[0300] Crystallization Method 7: A 500 pl aliquot of semaglutide (80 mg / ml), in water was mixed with 1000 pl of reagent containing 200 mM Zinc acetate, 100 mM imidazole / Hydrochloric acid pH 8.0, 20% w / v PEG 3000 and incubated at room temperature overnight. The final concentration of the semaglutide in solution was 26.67 mg / ml. Thismixture was then mixed using a votex and left at room temperature. Semaglutide Micro Particles were obtained on the following day. About 80% of the input semaglutide was formed microparticles by this method.

[0301] Crystallization Method 8: A 500 pl aliquot of semaglutide (80 mg / ml), in water was mixed with 1000 pl of reagent containing 2500 mM Sodium chloride, 100 mM imidazole / Hydrochloric acid pH 8.0, 200 mM zinc acetate and incubated at room temperature overnight. The final concentration of the semaglutide in solution was 26.67 mg / ml. This mixture was then mixed using a votex and left at room temperature. Semaglutide Micro Particles were obtained on the following day. About 85% of the input semaglutide was formed microparticles by this method.

[0302] Crystallization Method 9: A 500 pl aliquot of semaglutide (80 mg / ml), in water was mixed with 1000 pl of reagent containing 1300 mM Sodium chloride, 100 mM imidazole / Hydrochloric acid pH 6.5, 100 mM calcium chloride, 30% v / v isopropanol and incubated at room temperature overnight. The final concentration of the semaglutide in solution was 26.67 mg / ml. This mixture was then mixed using a votex and left at room temperature. Semaglutide Micro Particles were obtained on the following day. About 80% of the input semaglutide was formed microparticles by this method.

[0303] Crystallization Method 10: A 500 pl aliquot of semaglutide (80 mg / ml), in water was mixed with 1000 pl of reagent containing 871 mM Sodium chloride, 100 mM imidazole / Hydrochloric acid pH 6.5, 100 mM calcium chloride, 20.1% v / v isopropanol and incubated at room temperature overnight. The final concentration of the semaglutide in solution was 26.67 mg / ml. This mixture was then mixed using a votex and left at room temperature. Semaglutide Micro Particles were obtained on the following day. About 70% of the input semaglutide was formed microparticles by this method.

[0304] Crystallization Method 11: A 500 pl aliquot of semaglutide (80 mg / ml), in water was mixed with 1000 pl of reagent containing 9.9% v / v MPD, 100 mM sodium acetate / acetic acid acid pH 5.5, 100 mM calcium chloride, 4.95% PEG 8000 and incubated at room temperature overnight. The final concentration of the semaglutide in solution was 26.67 mg / ml. This mixture was then mixed using a votex and left at room temperature. Semaglutide Micro Particles were obtained on the following day. About 85% of the input semaglutide was formed microparticles by this method.

[0305] Crystallization Method 12: A 500 pl aliquot of semaglutide (80 mg / ml), in water was mixed with 1000 pl of reagent containing 10% v / v isopropanol, 100 mM imidazole / Hydrochloric acid pH 6.5, 100 mM calcium chloride, 30% v / v PEG 1500 and incubated at room temperature overnight. The final concentration of the semaglutide in solution was 26.67 mg / ml. This mixture was then mixed using a votex and left at room temperature. Semaglutide Micro Particles were obtained on the following day. About 90% of the input semaglutide was formed microparticles by this method.

[0306] Crystallization Method 13: A 500 pl aliquot of semaglutide (80 mg / ml), in water was mixed with 1000 pl of reagent containing 15% v / v isopropanol, 100 mM ammonium citrate / ammonium hydroxide pH 8.5 and incubated at room temperature overnight. The final concentration of the semaglutide in solution was 26.67 mg / ml. This mixture was then mixed using a votex and left at room temperature. Semaglutide Micro Particles were obtained on the following day. About 80% of the input semaglutide was formed microparticles by this method.

[0307] Crystallization Method 14: A 500 pl aliquot of semaglutide (80 mg / ml), in water was mixed with 1000 pl of reagent containing 2100 mM Sodium formate, 100 mM sodium acetate / acetic acid pH 4.5, 100 mM calcium chloride, 25% v / v PEG 3350 and incubated at room temperature overnight. The final concentration of the semaglutide in solution was 26.67 mg / ml. This mixture was then mixed using a votex and left at room temperature. Semaglutide Micro Particles were obtained on the following day. About 75% of the input semaglutide was formed microparticles by this method.Example 3: A comparative studies between commercially available drug, oral amorphous Semaglutide (Rybelsus®) Tablets, and Tablets containing crystalline Semaglutide with Vitamin E Formulation in Guinea pigs

[0308] The present Example demonstrates that oral formulations of Semaglutide exemplary of oral formulations disclosed herein have therapeutically effective pharmacokinetic profiles, e.g., with advantageous properties as compared to oral administration of the commercially available amorphous semaglutide Rybelsus® drug. The present example demonstrates a crystalline semaglutide Tablet formulation containing Vitamin E can be more effective than Rybelsus® containing intestinal permeation enhancer salcaprozate sodium (SNAC) (which is present as an intestinal permeation enhancer inRybelsus®) demonstrating that intestinal permeation enhancer SNAC is not required for advantageous application of formulations of the present disclosure.

[0309] The present Example describes an oral formulation of crystalline peptide / protein embedded in Tablet prepared as follows: Commercially obtained semaglutide (synthetic semaglutide) from PRIVEEL PEPTIDES is a lyophilized white powder (>95% purity). The semaglutide was processed to prepare crystalline microparticles containing semaglutide according to the procedure mentioned under Example 1. The crystalline microparticle of semaglutide was then lyophilized after washing with cold isopropanol. The amount of lyophilized semaglutide was quantified against a reference standard using a C18 reverse phase HPLC column (gradient elution, solvent A (0.1% TFA in water), solvent B (0.1% TFA in acetonitrile)). The lyophilized semaglutide (dosage mentioned under Table 2) was then used to prepare tablets containing following composition: The tablets were then stored at 4°C until further use.Table 3. Tablet composition of Semaglutide

[0310] Each 3 mg of Rybelsus® tablet contained magnesium stearate, microcrystalline cellulose, povidone along with 300 mg of intestinal absorption enhancer salcaprozate sodium (SNAC) and 3 mg of amorphous semaglutide.Table 4. Experimental Design for Pharmacokinetics Analysis

[0311] Pharmacokinetic analysis was conducted in male Guinea pigs weighing 300 to 400 grams for 4 days. Experimental design is shown in Table 2. Animals were acclimatized for 5-7 days under laboratory conditions. On the day of experiment all the animals were fasted overnight prior to dosing (food but not water withheld). Following the fasting period, the animals were weighed and the test items were administered via oral route as per the groupings. In the oral route of administration, animals were dosed 1 tablet / animal and tablets were suspended in 0.5% CMC. Blood samples were collected at aforementioned time points 0 min, 15 min, Ihr, 2hrs, 6hrs, 24 hrs, 48 hrs, 72 hrs and 96 hrs into an EDTA-blood collection tubes for the separation of plasma to determine Semaglutide levels by ELISA. Plasma samples collected at different time points were analyzed by ELISA kit according to Krishgen Biosystems protocol. This kit is based on sandwich enzyme-linked immuno -sorbent assay technology. An antibody, anti-semaglutide was pre-coated onto a 96-well plate.Standards, test samples, and biotin conjugated reagent were added to the wells and incubated. The HRP-conjugated reagent was then added, and the whole plate was incubated. Unbound conjugates were removed using wash buffer at each stage. TMB substrate was used to quantify the HRP enzymatic reaction. After TMB substrate was added, only wells that contain sufficient Semaglutide will produce a blue colored product, which then changes to yellow after adding the acidic stop solution. The intensity of the yellow color is proportional to the Semaglutide amount bound on the plate. The Optical Density (OD) was measuredspectrophotometrically at 450 nm in a microplate reader, from which the concentration of Semaglutide can be calculated.Table 5: Mean Semaglutide levels summary table (Guinea pigs) in ng / mlTable 6: Pharmacokinetic profile of Rybelsus® Tablet in Guinea pigsLambda_z 1 / h 0.032269991 tl / 2 h 21.47962116Tmax h 2Cmax ng / ml 1328.295CO ng / ml 14.59566667Clast_obs / Cmax 0.031522365AUC O-t ng / ml*h 31843.68958AUC 0-inf_obs ng / ml*h 33141.21086AUC 0-t / 0- inf_obs 0.960848706AUMC 0- inf_obs ng / ml*hA2 1008633.6MRT 0-inf_obs h 30.4344221Vz_obs (mg) / (ng / ml) 0.002805137Cl_obs (mg) / (ng / ml) / h 9.05217E-05Vss_obs (mg) / (ng / ml) 0.002754977Table 7: Pharmacokinetic profile of Crystalline Semaglutide Tablet with Vitamin E inGuinea pigsLambda_z 1 / h 0.0413005 tl / 2 h 16.78302167Tmax h 6Cmax ng / ml 4595.672333CO ng / ml 18.72666667Clast_obs / Cmax 0.037491141AUC O-t ng / ml*h 211970.0503AUC 0-inf_obs ng / ml*h 216141.84AUC 0-t / 0- inf_obs 0.980698833AUMC 0- inf_obs ng / ml*hA2 6636653.616MRT 0-inf_obs h 30.70508521Vz_obs (mg) / (ng / ml) 0.000336068Cl_obs (mg) / (ng / ml) / h 1.38798E-05Vss_obs (mg) / (ng / ml) 0.00042618

[0312] The data in Table 3 demonstrate average plasma concentration of Semaglutide at different time points. The pharmacokinetic profile are shown in Tables 4 and 5. Orally administered crystalline Semaglutide formulation with Vitamin E showed almost 7 fold increase in AUC when compared to Rybelsus®. Semaglutide was analyzed in plasma using a specific enzyme linked immunosorbent assay (ELISA). Pharmacokinetic endpoints were determined using noncompartmental methods using PK Solutions software. AUC was approximated using the trapezoidal rule on the observed concentrations. C max for each Semaglutide dose was derived as the maximum of all valid concentrations, and t max was then determined as the corresponding time point to C max. The terminal elimination rate constant (X z) was estimated by log-linear regression on the terminal part of the concentration-time curve, and t * was then calculated as t * = In 2 / A Z.Example 4: Comparative studies between oral Semaglutide Tablets containing crystalline semaglutide with Vitamin E vs Tablets containing crystalline Semaglutide without Vitamin E Formulation in Guinea pigs

[0313] The present Example demonstrates that oral formulations of crystalline Semaglutide with Vitamin E exemplary of oral formulations disclosed herein have therapeutically effective pharmacokinetic profiles, e.g., with advantageous properties as compared to oral administration of the crystalline semaglutide without Vitamin E. Thepresent example demonstrates a crystalline semaglutide Tablet formulation containing Vitamin E can be more effective than crystalline semaglutide formulation without Vitamin E formulation demonstrating that the Vitamin E is required for advantageous application of formulations of the present disclosure.

[0314] The present Example includes an oral formulation of crystalline peptide / protein embedded in Tablet prepared as follows: Commercially obtained semaglutide (synthetic semaglutide) from PRIVEEL PEPTIDES is a lyophilized white powder (>95% purity). The semaglutide was processed to prepare crystalline microparticles containing semaglutide according to the procedure mentioned under Example 1. The crystalline microparticle of semaglutide was then lyophilized after washing with cold isopropanol. The amount of lyophilized semaglutide was quantified against a reference standard using a C18 reverse phase HPLC column (gradient elution, solvent A (0.1% TFA in water), solvent B (0.1% TFA in acetonitrile)). The lyophilized semaglutide (dosage mentioned under Table 8) was then used to prepare tablets containing following composition: The tablets were then stored at 4°C until further use.Table 8. Tablet composition of Crystalline Semaglutide with Vitamin E

[0315] Each 3 mg of crystalline semaglutide used for making tablet without VitaminE contains following formulation as follows:Table 9. Tablet composition of Crystalline Semaglutide without Vitamin ETable 10. Experimental Design for Pharmacokinetics Analysis

[0316] Pharmacokinetic analysis was conducted in male Guinea pigs weighing 300 to 400 grams for 4 days. Experimental design is shown in Table 8. Animals were acclimatized for 5-7 days under laboratory conditions. On the day of experiment all the animals were fasted overnight prior to dosing (food but not water withheld). Following the fasting period, the animals were weighed and the test items were administered via oral route as per the groupings. In the oral route of administration, animals were dosed 1 tablet / animal and tablets were suspended in 0.5% CMC. Blood samples were collected at aforementioned time points 0 min, 15 min, Ihr, 2hrs, 6hrs, 24 hrs, 48 hrs, 72 hrs and 96 hrs into an EDTA-blood collection tubes for the separation of plasma to determine Semaglutide levels by ELISA. Plasma samples collected at different time points were analyzed by ELISA kit according to Krishgen Biosystems protocol. This kit is based on sandwich enzyme-linked immuno -sorbent assay technology. An antibody, anti-semaglutide was pre-coated onto a 96-well plate.Standards, test samples, and biotin conjugated reagent were added to the wells and incubated. The HRP-conjugated reagent was then added, and the whole plate was incubated. Unbound conjugates were removed using wash buffer at each stage. TMB substrate was used to quantify the HRP enzymatic reaction. After TMB substrate was added, only wells that contain sufficient Semaglutide will produce a blue colored product, which then changes to yellow after adding the acidic stop solution. The intensity of the yellow color is proportional to the Semaglutide amount bound on the plate. The Optical Density (OD) was measured spectrophotometrically at 450 nm in a microplate reader, from which the concentration of Semaglutide can be calculated.Table 11: Mean Semaglutide levels summary table (Guinea pigs) in ng / mlTable 12: Pharmacokinetic profile of Crystalline Semaglutide Tablet without Vitamin E in Guinea pigsLambda_z 1 / h 0.009478012 tl / 2 h 73.13212382Tmax h 2Cmax ng / ml 291.7913333CO ng / ml 35.67633333Clast_obs / Cmax 0.497091529AUC 0-t ng / ml*h 17080.87158AUC 0-inf_obs ng / ml*h 32384.39652AUC 0-t / 0- inf_obs 0.527441404AUMC 0-inf_obs ng / ml*hA2 3769632.412MRT 0-inf_obs h 116.4027377Vz_obs (mg) / (ng / ml) 0.009773906Cl_obs (mg) / (ng / ml) / h 9.26372E-05Vss_obs (mg) / (ng / ml) 0.010783224Table 13: Pharmacokinetic profile of Crystalline Semaglutide Tablet with Vitamin E in Guinea pigsLambda_z 1 / h 0.0413005 tl / 2 h 16.78302167Tmax h 6Cmax ng / ml 4595.672333CO ng / ml 18.72666667Clast_obs / Cmax 0.037491141AUC O-t ng / ml*h 211970.0503AUC 0-inf_obs ng / ml*h 216141.84AUC 0-t / 0- inf_obs 0.980698833AUMC 0- inf_obs ng / ml*hA2 6636653.616MRT 0-inf_obs h 30.70508521Vz_obs (mg) / (ng / ml) 0.000336068Cl_obs (mg) / (ng / ml) / h 1.38798E-05Vss_obs (mg) / (ng / ml) 0.00042618

[0317] The data in Table 9 demonstrate average plasma concentration of Semaglutide at different time points. The full pharmacokinetic profile are shown in Tables 10 and 11. Orally administered crystalline Semaglutide formulation without Vitamin E showed lower AUC when compared with one containing Vitamin E formulation. Semaglutide was analyzed in plasma using a specific enzyme linked immunosorbent assay (ELISA). Pharmacokinetic endpoints were determined using noncompartmental methods using PK Solutions software. AUC was approximated using the trapezoidal rule on the observed concentrations. C max for each Semaglutide dose was derived as the maximum of all valid concentrations, and t max was then determined as the corresponding time point to C max. The terminal elimination rate constant (X z) was estimated by log-linear regression on the terminal part of the concentration-time curve, and t * was then calculated as t * = In 2 / A Z.Example 5: Comparative studies between oral Semaglutide Tablets containing crystalline semaglutide with Vitamin E vs Tablets containing commercial amorphous Semaglutide with Vitamin E Formulation in Guinea pigs

[0318] The present Example demonstrates that oral formulations of crystalline Semaglutide with Vitamin E exemplary of oral formulations disclosed herein have therapeutically effective pharmacokinetic profiles, e.g., with advantageous properties as compared to oral administration of the commercially available amorphous semaglutide with Vitamin E. The present example demonstrates a crystalline semaglutide Tablet formulation containing Vitamin E can be more effective than amorphous semaglutide formulation with Vitamin E formulation demonstrating that crystalline semaglutide is required for advantageous application of formulations of the present disclosure.

[0319] The present Example includes an oral formulation of crystalline peptide / protein embedded in Tablet prepared as follows: Commercially obtained semaglutide (synthetic semaglutide) from PRIVEEL PEPTIDES is a lyophilized white powder (>95% purity). The semaglutide was processed to prepare crystalline microparticles containing semaglutide according to the procedure mentioned under Example 1. The crystalline microparticle of semaglutide was then lyophilized after washing with cold isopropanol. The amount of lyophilized semaglutide was quantified against a reference standard using a C18 reverse phase HPLC column (gradient elution, solvent A (0.1% TFA in water), solvent B (0.1% TFA in acetonitrile)). The lyophilized semaglutide (dosage mentioned under Table 14) was then used to prepare tablets containing following composition: The tablets were then stored at 4°C until further use.Table 14. Tablet composition of Crystalline Semaglutide with Vitamin E

[0320] Each 3 mg of commercially available amorphous semaglutide used for making tablet containing magnesium stearate, microcrystalline cellulose, povidone, Vitamin E and 3 mg of amorphous semaglutide as follows.Table 15. Tablet composition of Crystalline Semaglutide with Vitamin ETable 16. Experimental Design for Pharmacokinetics Analysis

[0321] Pharmacokinetic analysis was conducted in male Guinea pigs weighing 300 to 400 grams for 4 days. Experimental design is shown in Table 14. Animals were acclimatized for 5-7 days under laboratory conditions. On the day of experiment all the animals were fasted overnight prior to dosing (food but not water withheld). Following the fasting period, the animals were weighed and the test items were administered via oral route as per the groupings. In the oral route of administration, animals were dosed 1 tablet / animal and tablets were suspended in 0.5% CMC. Blood samples were collected at aforementioned time points 0 min, 15 min, Ihr, 2hrs, 6hrs, 24 hrs, 48 hrs, 72 hrs and 96 hrs into an EDTA-blood collection tubes for the separation of plasma to determine Semaglutide levels by ELISA. Plasma samples collected at different time points were analyzed by ELISA kit according to Krishgen Biosystems protocol. This kit is based on sandwich enzyme-linked immuno -sorbent assay technology. An antibody, anti-semaglutide was pre-coated onto a 96-well plate.Standards, test samples, and biotin conjugated reagent were added to the wells and incubated. The HRP-conjugated reagent was then added, and the whole plate was incubated. Unbound conjugates were removed using wash buffer at each stage. TMB substrate was used to quantify the HRP enzymatic reaction. After TMB substrate was added, only wells that contain sufficient Semaglutide will produce a blue colored product, which then changes to yellow after adding the acidic stop solution. The intensity of the yellow color is proportional to the Semaglutide amount bound on the plate. The Optical Density (OD) was measured spectrophotometrically at 450 nm in a microplate reader, from which the concentration of Semaglutide can be calculated.Table 17: Mean Semaglutide levels summary table (Guinea pigs) in ng / mlTable 18: Pharmacokinetic profile of amorpohous Semaglutide Tablet with Vitamin E inGuinea pigsLambda.z 1 / h 0.023285093 tl / 2 h 29.76785082Tmax h 6Cmax ng / ml 591.589CO ng / ml 48.06166667Clast_obs / Cmax 0.12686116AUC 0-t ng / ml*h 24472.22867AUC 0-inf_obs ng / ml*h 27695.30645AUC 0-t / 0- inf_obs 0.883623682AUMC 0- inf_obs ng / ml*hA2 1190929.327MRT 0-inf_obs h 43.00112471Vz_obs (mg) / (ng / ml) 0.004651972Cl_obs (mg) / (ng / ml) / h 0.000108322Vss_obs (mg) / (ng / ml) 0.004657951Table 19: Pharmacokinetic profile of crystalline Semaglutide Tablet with Vitamin E in Guinea pigsLambda_z 1 / h 0.0413005 tl / 2 h 16.78302167Tmax h 6Cmax ng / ml 4595.672333CO ng / ml 18.72666667Clast_obs / Cmax 0.037491141AUC 0-t ng / ml*h 211970.0503AUC 0-inf_obs ng / ml*h 216141.84AUC 0-t / 0- inf_obs 0.980698833AUMC 0- inf_obs ng / ml*hA2 6636653.616MRT 0-inf_obs h 30.70508521Vz_obs (mg) / (ng / ml) 0.000336068Cl_obs (mg) / (ng / ml) / h 1.38798E-05Vss_obs _ (mg) / (ng / ml) 0.00042618

[0322] The data in Table 15 demonstrate average plasma concentration of Semaglutide at different time points. The full pharmacokinetic profile are shown in Tables 16 and 17. Orally administered crystalline Semaglutide formulation showed 9 fold increase in AUC when compared to amorphous semaglutide formulation with Vitamin E. Semaglutide was analyzed in plasma using a specific enzyme linked immunosorbent assay (ELISA). Pharmacokinetic endpoints were determined using noncompartmental methods using PK Solutions software. AUC was approximated using the trapezoidal rule on the observed concentrations. C max for each Semaglutide dose was derived as the maximum of all valid concentrations, and t max was then determined as the corresponding time point to C max. The terminal elimination rate constant (X z) was estimated by log-linear regression on the terminal part of the concentration-time curve, and t * was then calculated as t * = In 2 / A z.Example 6: Pharmacokinetic Study of Semaglutide Formulation Enclosed in Fast Dissolving Tablets or capsule with Vitamin E in mini pigs

[0323] The present Example demonstrates that oral formulations of crystalline Semaglutide exemplary of oral formulations disclosed herein have therapeutically effective pharmacokinetic profiles, e.g., with advantageous properties as compared to oral administration of the commercially available Rybelsus® Tablets. The present example demonstrates either a crystalline semaglutide fast dissolving Tablet formulation or a capsule formulation with Vitamin E can be more effective than Rybelsus® demonstrating that crystalline semaglutide either in fast dissolving tablet form for sublingual delivery or in capsule form with Vitamin E is required for advantageous application of formulations of the present disclosure.

[0324] The present Example describes an oral formulation of crystalline peptide / protein embedded in Tablet prepared as follows: Commercially obtained semaglutide (synthetic semaglutide) from PRIVEEL PEPTIDES is a lyophilized white powder (>95% purity). The semaglutide was processed to prepare crystalline microparticles containing semaglutide according to the procedure mentioned under Example 1. The crystalline microparticle of semaglutide was then lyophilized after washing with cold isopropanol. The amount of lyophilized semaglutide was quantified against a reference standard using a C18 reverse phase HPLC column (gradient elution, solvent A (0.1% TFA in water), solvent B(0.1% TFA in acetonitrile)). The lyophilized semaglutide (dosage mentioned under Table 20)was then used to prepare tablets containing following composition: The tablets were then stored at 4°C until further use.Table 20. Tablet composition of Fast Dissolving crystalline Semaglutide TabletTable 21. Capsule composition of crystalline SemaglutideTable 22. Experimental Design for Pharmacokinetics Analysis

[0325] Pharmacokinetic analysis was conducted in female mini pigs weighing 11+1 kgs for 7 days. Experimental design is shown in Table 20. Animals were acclimatized for 5-7 days under laboratory conditions. On the day of experiment all the animals were fasted overnight prior to dosing (food but not water withheld). Following the fasting period, the animals were weighed and the test items were administered via SC or IV or oral route as per the groupings. In the oral route of administration, animals were dosed 1 tablet / animal and tablets were suspended in 0.5% CMC. Blood samples were collected at aforementioned time points 0 hr, 0.5 hr, 1 hr, 2 hrs, 4hrs, 6 hrs, 8 hrs, 12 hrs, 24 hrs, 48 hrs, 96 hrs, 120 hrs, 144 hrs and 168 hrs into an EDTA-blood collection tubes for the separation of plasma to determine Semaglutide levels by ELISA. Plasma samples collected at different time points were analyzed by ELISA kit according to Krishgen Biosystems protocol. This kit is based on sandwich enzyme-linked immuno- sorbent assay technology. An antibody, anti-semaglutide was pre-coated onto a 96-well plate. Standards, test samples, and biotin conjugated reagent were added to the wells and incubated. The HRP-conjugated reagent was then added, and the whole plate was incubated. Unbound conjugates were removed using wash buffer at each stage. TMB substrate was used to quantify the HRP enzymatic reaction. After TMB substrate was added, only wells that contain sufficient Semaglutide will produce a blue colored product, which then changes to yellow after adding the acidic stop solution. The intensity of the yellow color is proportional to the Semaglutide amount bound on the plate. The Optical Density (OD) was measured spectrophotometrically at 450 nm in a microplate reader, from which the concentration of Semaglutide can be calculated.Table 23: Subcutaneous PK profile of Ozempic injectionTable 24: Full PK analysis of SC administration of Ozempic injectionTable 25: Intravenous PK profile of Ozempic injectionTable 26: Full PK analysis of intravenous administration of Ozempic injectionTable 27: Oral PK profile of Rybelsus tablet (amorphous semaglutide)Table 28: Sublingual PK profile of Crystalline semaglutide tabletTable 29: Full PK analysis oral administration of Rybelsus and Crystalline semaglutide tabletTable 30: Oral PK profile of Crystalline semaglutide capsuleTable 31: Full PK analysis oral administration of Crystalline semaglutide capsule

[0326] The data in Tables 21, 23, 25, 26 and 28 demonstrate average plasma concentration of Semaglutide at different time points. The full pharmacokinetic profile are shown in Tables 22, 24, 27 and 29. Orally administered crystalline Semaglutide formulation either through sublingual route or in capsule form orally showed better pharmacokinetic profile than Rybelsus®. Semaglutide was analyzed in plasma using a specific enzyme linked immunosorbent assay (ELISA). Pharmacokinetic endpoints were determined using noncompartmental methods using PK Solutions software. AUC was approximated using the trapezoidal rule on the observed concentrations. C max for each Semaglutide dose was derived as the maximum of all valid concentrations, and t max was then determined as the corresponding time point to C max. The terminal elimination rate constant (X z) wasestimated by log-linear regression on the terminal part of the concentration-time curve, and t * was then calculated as t * = In 2 / A z.Example 7: Preparation of Fast Dissolving Tablets or capsule containing sugars or albumin or amylase - semaglutide

[0327] The present Example descibes an oral formulation of crystalline peptide / protein embedded in Tablet is prepared as follows: Commercially available semaglutide (synthetic semaglutide) from PRIVEEL PEPTIDES is a lyophilized white powder (>95% purity). The semaglutide is processed to prepare crystalline microparticles containing semaglutide with sugar / sugars (such as Trehalose, mannitol, sorbitol, sucrose, mannose etc. either alone or in combination of various sugars) or with proteins (such as albumin or amylase) according to the procedure mentioned under Example 1. The crystalline microparticle of semaglutide containing sugar or protein is then lyophilized after washing with cold isopropanol. The amount of lyophilized semaglutide is quantified against a reference standard using a C18 reverse phase HPLC column (gradient elution, solvent A (0.1% TFA in water), solvent B (0.1% TFA in acetonitrile)). The lyophilized semaglutide containing either sugar or protein is then used to prepare tablets or capsules containing following composition: The tablets or capsules are then stored at 4°C until further use.Table 32. Tablet composition of Fast Dissolving crystalline Semaglutide TabletTable 33. Capsule composition of crystalline SemaglutideExample 8: Preparation of Tablets containing sugars or albumin or amylase - semaglutide

[0328] The present Example includes an oral formulation of crystalline peptide / protein embedded in Tablet is prepared as follows: Commercially obtained semaglutide (synthetic semaglutide) from PRIVEEL PEPTIDES is a lyophilized white powder (>95% purity). The semaglutide is processed to prepare crystalline microparticles containing semaglutide with sugar / sugars (such as Trehalose, mannitol, sorbitol, sucrose, mannose etc. either alone or in combination of various sugars) or with proteins (such as albumin or amylase) according to the procedure mentioned under Example 1. The crystalline microparticle of semaglutide containing sugar or protein is then lyophilized after washing with cold isopropanol. The amount of lyophilized semaglutide is quantified against a reference standard using a C18 reverse phase HPLC column (gradient elution, solvent A (0.1% TFA in water), solvent B (0.1% TFA in acetonitrile)). The lyophilized semaglutide containing either sugar or protein is then used to prepare tablets or capsules containing following composition: The tablets or capsules are then stored at 4°C until further use.Table 34. Tablet composition of Crystalline Semaglutide with Vitamin ETable 35. Tablet composition of Crystalline Semaglutide without Vitamin EExample 9: Crystallization of Albumin Carrier Protein Containing Semaglutide

[0329] The present Example demonstrates formation of semaglutide embedded in crystallized albumin for use in an oral formulation as disclosed herein.

[0330] Semaglutide is supplied as one ml of solution which contains 1.34 mg of semaglutide. One pre-filled pen contains 2 mg Semaglutide in 1.5 ml solution. Each dose contains 0.25 mg of Semaglutide in 0.19 ml solution. Oral Semaglutide is a tablet formulation for once-daily administration. It is co-formulated with an absorption enhancer, sodium N- (8- [2-hydroxybenzoyl] amino) caprylate (SNAC). The present Example includes the recognitionthat Semaglutide is exemplary of peptide / proteins for which an oral formulation would be advantageous.

[0331] The present Example utilizes Semaglutide. To produce albumin carrier protein containing Semaglutide microparticles, Semaglutide in the lyophilized form is dissolved in water at a concentration of 80 mg / mL. Semaglutide aliquots are then mixed with human albumin (20% solution of ALBUCEL® - Human Albumin - 20 g / 100 mL, INTAS) solution to make albumin crystals containing Semaglutide using methods described below.

[0332] Crystallization Method for albumin containing embedded Semaglutide: A 500 pL aliquot of albumin (-200 mg / mL), in water is mixed 500 pL of Semaglutide solution (~ 80 mg / mL) with 1000 pL of reagent containing 20% (w / v) PEG 3350, and is incubated at room temperature overnight. The final concentration of the Semaglutide in solution is 40 mg / mL. This mixture is then mixed using a votex mixer and is left at room temperature. Albumin microparticles containing Semaglutide are obtained on the following day. About 90% of the input albumin is formed microparticles containing embedded Semaglutide by this method.

[0333] In order to estimate the amount of Semaglutide embedded in albumin crystals, the albumin crystals containing Semaglutide are harvested by centrifugation of the sample at 4000 x g. The supernatant is discarded. The crystals are then washed twice with 1 mL of albumin crystallization reagent (20% (w / v) PEG 3350). The washed crystals are then lyophilized after the final wash with cold isopropanol. The amount of Semaglutide in the lyophilized sample is then quantified against a reference standard using a C18 reverse phase 30 HPLC column (gradient elution, solvent A (0.1% TFA in water), solvent B (0.1% TFA in acetonitrile)). About 8 to 10% of the Semaglutide is getting embedded / trapped inside the lyophilized albumin crystals.Example 10: Crystallization of Amylase Carrier Protein Containing Semaglutide

[0334] The present Example demonstrates formation of crystallized amylase carrier protein embedded with semaglutide for use in an oral formulation as disclosed herein. The present Example includes the recognition that semaglutide is exemplary of peptide / proteins for which an oral formulation would be advantageous.

[0335] The present Example utilizes Semaglutide. To produce amylase carrier protein containing Semaglutide microparticles, Semaglutide in the lyophilized form is dissolved in water at a concentration of 80 mg / mL. Semaglutide aliquots are then mixed with amylasefrom Aspergillus oryzae (Sigma Chemical Company) solution to make amylase crystals containing semaglutide using methods described below.

[0336] Crystallization method for amylase containing embedded Semaglutide: A 500 pL aliquot of amylase (-200 mg / mL), in water is mixed 500 pL of semaglutide solution (~ 80 mg / mL) with 1000 pL of reagent containing 30% (w / v) PEG 8000, 100 mM Sodium acetate / Acetic acid pH 4.5 and 200 mM Lithium sulfate, and is incubated at room temperature overnight. The final concentration of the semaglutide in solution is 20 mg / mL. This mixture is then mixed using a votex and is left at room temperature. Amylase microparticles containing semaglutide are obtained on the following day. About 90% of the input amylase is formed microparticles containing embedded semaglutide by this method.

[0337] In order to estimate the amount of semaglutide embedded in amylase crystals, the amylase crystals containing semaglutide are harvested by centrifugation of the sample at 4000 x g. The supernatant is discarded. The crystals are then washed twice with 1 mL of amylase crystallization reagent (30% (w / v) PEG 8000, 100 mM sodium acetate / acetic acid pH 4.5 and 200 mM Lithium sulfate). The washed crystals are then lyophilized after the final wash with cold isopropanol. The amount of semaglutide in the lyophilized sample is then quantified against a reference standard using a Cl 8 reverse phase HPLC column (gradient elution, solvent A (0.1% TFA in water), solvent B (0.1% TFA in acetonitrile)). About 8 to 10% of the semaglutide is getting embedded / trapped inside the lyophilized amylase crystals.Example 11. Crystallization of Semaglutide

[0338] The present Example demonstrates formation of crystallized semaglutide for use in an oral formulation as disclosed herein. Semaglutide, sold under the brand names Wegovy® and Ozempic® among others, is an antidiabetic medication used for the treatment of type 2 diabetes and long-term weight management. Semaglutide is a GLP-1 receptor agonist, meaning that it mimics the action of the human incretin glucagon-like peptide- 1 (GLP-1), thereby increasing insulin secretion and increasing blood sugar disposal and improving glycemic control. Semaglutide is indicated as an adjunct to diet and exercise to improve glycemic control in adults with type 2 diabetes. Semaglutide is also indicated as an adjunct to diet and exercise for long-term weight management in adults with obesity. The subcutaneous injection is administered once weekly and the tablet is administered once a day. Semaglutide offers a competitive advantage over other drugs used to manage diabetes, which may require several daily doses.

[0339] Semaglutide is supplied as one ml of solution which contains 1.34 mg of semaglutide. One pre-filled pen contains 2 mg Semaglutide in 1.5 ml solution. Each dose contains 0.25 mg of Semaglutide in 0.19 ml solution. Oral Semaglutide is a tablet formulation for once-daily administration. It is co-formulated with an absorption enhancer, sodium N- (8- [2-hydroxybenzoyl] amino) caprylate (SNAC). The present Example includes the recognition that Semaglutide is exemplary of peptide / proteins for which an oral formulation would be advantageous.

[0340] The present Example utilizes semaglutide. To produce semaglutide microparticles, semaglutide solution was dialyzed against water for 24 hr with three changes and lyophilized to get dry powder. The lyophilized powder of semaglutide was then dissolved in water at a concentration of 80 mg / mL. Semaglutide aliquots were then crystalized using a variety of methods described below.

[0341] Crystallization Method 1:

[0342] A 500 pl aliquot of semaglutide (80 mg / ml), in water was mixed with 1000 pl of reagent containing 50 mM Magnesium chloride, 100 mM Tis / Hydrochloric acid pH 8.5, 40% v / v reagent alcohol and incubated at room temperature overnight. The final concentration of the semaglutide in solution was 26.67 mg / ml. This mixture was then mixed using a votex and left at room temperature. Semaglutide Micro Particles were obtained on the following day. About 80% of the input semaglutide was formed microparticles by this method.

[0343] Crystallization Method 2:

[0344] A 500 pl aliquot of semaglutide (80 mg / ml), in water was mixed with 1000 pl of reagent containing 200 mM Magnesium chloride, 100 mM sodium cacodylate / Hydrochloric acid pH 6.5, 10% w / v PEG 3000 and incubated at room temperature overnight. The final concentration of the semaglutide in solution was 26.67 mg / ml. This mixture was then mixed using a votex and left at room temperature. Semaglutide Micro Particles were obtained on the following day. About 70% of the input semaglutide was formed microparticles by this method.

[0345] Crystallization Method 3:

[0346] A 500 pl aliquot of semaglutide (80 mg / ml), in water was mixed with 1000 pl of reagent containing 200 mM Magnesium chloride, 100 mM Tris / Hydrochloric acid pH 7.0, 10% w / v PEG 8000 and incubated at room temperature overnight. The final concentration ofthe semaglutide in solution was 26.67 mg / ml. This mixture was then mixed using a votex and left at room temperature. Semaglutide Micro Particles were obtained on the following day. About 70% of the input semaglutide was formed microparticles by this method.Crystallization

[0347] Crystallization Method 4:

[0348] A 500 pl aliquot of semaglutide (80 mg / ml), in water was mixed with 1000 pl of reagent containing 200 mM Magnesium chloride, 100 mM sodium cacodylate / Hydrochloric acid pH 6.5, 20% w / v PEG 1000 and incubated at room temperature overnight. The final concentration of the semaglutide in solution was 26.67 mg / ml. This mixture was then mixed using a votex and left at room temperature. Semaglutide Micro Particles were obtained on the following day. About 70% of the input semaglutide was formed microparticles by this method.

[0349] Crystallization Method 5:

[0350] A 500 pl aliquot of semaglutide (80 mg / ml), in water was mixed with 1000 pl of reagent containing 200 mM Sodium chloride, 100 mM sodium phosphate dibasic / potassium phosphate monobasic pH 6.2, 40% w / v PEG 400 and incubated at room temperature overnight. The final concentration of the semaglutide in solution was 26.67 mg / ml. This mixture was then mixed using a votex and left at room temperature. Semaglutide Micro Particles were obtained on the following day. About 90% of the input semaglutide was formed microparticles by this method.

[0351] Crystallization Method 6:

[0352] A 500 pl aliquot of semaglutide (80 mg / ml), in water was mixed with 1000 pl of reagent containing 200 mM Sodium chloride, 100 mM sodium phosphate dibasic / potassium phosphate monobasic pH 6.2, 35% v / v ethoxyethanol and incubated at room temperature overnight. The final concentration of the semaglutide in solution was 26.67 mg / ml. This mixture was then mixed using a votex and left at room temperature. Semaglutide Micro Particles were obtained on the following day. About 85% of the input semaglutide was formed microparticles by this method.

[0353] Crystallization Method 7:

[0354] A 500 pl aliquot of semaglutide (80 mg / ml), in water was mixed with 1000 pl of reagent containing 200 mM Zinc acetate, 100 mM imidazole / Hydrochloric acid pH 8.0,20% w / v PEG 3000 and incubated at room temperature overnight. The final concentration of the semaglutide in solution was 26.67 mg / ml. This mixture was then mixed using a votex and left at room temperature. Semaglutide Micro Particles were obtained on the following day. About 80% of the input semaglutide was formed microparticles by this method.

[0355] Crystallization Method 8:

[0356] A 500 pl aliquot of semaglutide (80 mg / ml), in water was mixed with 1000 pl of reagent containing 2500 mM Sodium chloride, 100 mM imidazole / Hydrochloric acid pH 8.0, 200 mM zinc acetate and incubated at room temperature overnight. The final concentration of the semaglutide in solution was 26.67 mg / ml. This mixture was then mixed using a votex and left at room temperature. Semaglutide Micro Particles were obtained on the following day. About 85% of the input semaglutide was formed microparticles by this method.

[0357] Crystallization Method 9:

[0358] A 500 pl aliquot of semaglutide (80 mg / ml), in water was mixed with 1000 pl of reagent containing 1300 mM Sodium chloride, 100 mM imidazole / Hydrochloric acid pH 6.5, 100 mM calcium chloride, 30% v / v isopropanol and incubated at room temperature overnight. The final concentration of the semaglutide in solution was 26.67 mg / ml. This mixture was then mixed using a votex and left at room temperature. Semaglutide Micro Particles were obtained on the following day. About 80% of the input semaglutide was formed microparticles by this method.

[0359] Crystallization Method 10:

[0360] A 500 pl aliquot of semaglutide (80 mg / ml), in water was mixed with 1000 pl of reagent containing 871 mM Sodium chloride, 100 mM imidazole / Hydrochloric acid pH 6.5, 100 mM calcium chloride, 20.1% v / v isopropanol and incubated at room temperature overnight. The final concentration of the semaglutide in solution was 26.67 mg / ml. This mixture was then mixed using a votex and left at room temperature. Semaglutide Micro Particles were obtained on the following day. About 70% of the input semaglutide was formed microparticles by this method.

[0361] Crystallization Method 11:

[0362] A 500 pl aliquot of semaglutide (80 mg / ml), in water was mixed with 1000 pl of reagent containing 9.9% v / v MPD, 100 mM sodium acetate / acetic acid acid pH 5.5, 100mM calcium chloride, 4.95% PEG 8000 and incubated at room temperature overnight. The final concentration of the semaglutide in solution was 26.67 mg / ml. This mixture was then mixed using a votex and left at room temperature. Semaglutide Micro Particles were obtained on the following day. About 85% of the input semaglutide was formed microparticles by this method.

[0363] Crystallization Method 12:

[0364] A 500 pl aliquot of semaglutide (80 mg / ml), in water was mixed with 1000 pl of reagent containing 10% v / v isopropanol, 100 mM imidazole / Hydrochloric acid pH 6.5, 100 mM calcium chloride, 30% v / v PEG 1500 and incubated at room temperature overnight. The final concentration of the semaglutide in solution was 26.67 mg / ml. This mixture was then mixed using a votex and left at room temperature. Semaglutide Micro Particles were obtained on the following day. About 90% of the input semaglutide was formed microparticles by this method.

[0365] Crystallization Method 13:

[0366] A 500 pl aliquot of semaglutide (80 mg / ml), in water was mixed with 1000 pl of reagent containing 15% v / v isopropanol, 100 mM ammonium citrate / ammonium hydroxide pH 8.5 and incubated at room temperature overnight. The final concentration of the semaglutide in solution was 26.67 mg / ml. This mixture was then mixed using a votex and left at room temperature. Semaglutide Micro Particles were obtained on the following day. About 80% of the input semaglutide was formed microparticles by this method.

[0367] Crystallization Method 14:

[0368] A 500 pl aliquot of semaglutide (80 mg / ml), in water was mixed with 1000 pl of reagent containing 2100 mM Sodium formate, 100 mM sodium acetate / acetic acid pH 4.5, 100 mM calcium chloride, 25% v / v PEG 3350 and incubated at room temperature overnight. The final concentration of the semaglutide in solution was 26.67 mg / ml. This mixture was then mixed using a votex and left at room temperature. Semaglutide Micro Particles were obtained on the following day. About 75% of the input semaglutide was formed microparticles by this method.Example 12: A comparative studies between commercially available drug, oral amorphous Semaglutide (Rybelsus®) Tablets, and Tablets containing crystalline Semaglutide with Vitamin E Formulation in Guinea pigs

[0369] The present Example demonstrates that oral formulations of Semaglutide exemplary of oral formulations disclosed herein have therapeutically effective pharmacokinetic profiles, e.g., with advantageous properties as compared to oral administration of the commercially available amorphous semaglutide Rybelsus® drug. The present example demonstrates a crystalline semaglutide Tablet formulation containing Vitamin E can be more effective than Rybelsus® containing intestinal permeation enhancer salcaprozate sodium (SNAC) (which is present as an intestinal permeation enhancer in Rybelsus®) demonstrating that intestinal permeation enhancer SNAC is not required for advantageous application of formulations of the present disclosure.

[0370] The present Example includes an oral formulation of crystalline peptide / protein embedded in Tablet prepared as follows: Commercially obtained semaglutide (synthetic semaglutide) from PRIVEEL PEPTIDES is a lyophilized white powder (>95% purity). The semaglutide was processed to prepare crystalline microparticles containing semaglutide according to the procedure mentioned under Example 1. The crystalline microparticle of semaglutide was then lyophilized after washing with cold isopropanol. The amount of lyophilized semaglutide was quantified against a reference standard using a C18 reverse phase HPLC column (gradient elution, solvent A (0.1% TFA in water), solvent B (0.1% TFA in acetonitrile)). The lyophilized semaglutide (dosage mentioned under Table 2) was then used to prepare tablets containing following composition: The tablets were then stored at 4°C until further use.Table 36. Tablet composition of Semaglutide

[0371] Each 3 mg of Rybelsus® tablet contained magnesium stearate, microcrystalline cellulose, povidone along with 300 mg of intestinal absorption enhancer salcaprozate sodium (SNAC) and 3 mg of amorphous semaglutide.Table 37. Experimental Design for Pharmacokinetics Analysis

[0372] Pharmacokinetic analysis was conducted in male Guinea pigs weighing 300 to 400 grams for 4 days. Experimental design is shown in Table 2. Animals were acclimatized for 5-7 days under laboratory conditions. On the day of experiment all the animals were fasted overnight prior to dosing (food but not water withheld). Following the fasting period, the animals were weighed and the test items were administered via oral route as per the groupings. In the oral route of administration, animals were dosed 1 tablet / animal and tablets were suspended in 0.5% CMC. Blood samples were collected at aforementioned time points 0 min, 15 min, Ihr, 2hrs, 6hrs, 24 hrs, 48 hrs, 72 hrs and 96 hrs into an EDTA-blood collection tubes for the separation of plasma to determine Semaglutide levels by ELISA. Plasma samples collected at different time points were analyzed by ELISA kit according to Krishgen Biosystems protocol. This kit is based on sandwich enzyme-linked immuno -sorbent assay technology. An antibody, anti-semaglutide was pre-coated onto a 96-well plate.Standards, test samples, and biotin conjugated reagent were added to the wells and incubated. The HRP-conjugated reagent was then added, and the whole plate was incubated. Unbound conjugates were removed using wash buffer at each stage. TMB substrate was used toquantify the HRP enzymatic reaction. After TMB substrate was added, only wells that contain sufficient Semaglutide will produce a blue colored product, which then changes to yellow after adding the acidic stop solution. The intensity of the yellow color is proportional to the Semaglutide amount bound on the plate. The Optical Density (OD) was measured spectrophotometrically at 450 nm in a microplate reader, from which the concentration of Semaglutide can be calculated.Table 38: Mean Semaglutide levels summary table (Guinea pigs) in ng / mlTable 39: Pharmacokinetic profile of Rybelsus® Tablet in Guinea pigsLambda.z 1 / h 0.032269991 tl / 2 h 21.47962116Tmax h 2Cmax ng / ml 1328.295CO ng / ml 14.59566667Clast_obs / Cmax 0.031522365AUC O-t ng / ml*h 31843.68958AUC 0-inf_obs ng / ml*h 33141.21086AUC 0-t / 0- inf_obs 0.960848706AUMC 0- inf_obs ng / ml*hA2 1008633.6MRT 0-inf_obs h 30.4344221Vz_obs (mg) / (ng / ml) 0.002805137Cl_obs (mg) / (ng / ml) / h 9.05217E-05Vss obs (mg) / (ng / ml) 0.002754977Table 40: Pharmacokinetic profile of Crystalline Semaglutide Tablet with vitamin E inGuinea pigsLambda_z 1 / h 0.0413005 tl / 2 h 16.78302167Tmax h 6Cmax ng / ml 4595.672333CO ng / ml 18.72666667Clast_obs / Cmax 0.037491141AUC O-t ng / ml*h 211970.0503AUC 0-inf_obs ng / ml*h 216141.84AUC 0-t / 0- inf_obs 0.980698833AUMC 0- inf_obs ng / ml*hA2 6636653.616MRT 0-inf_obs h 30.70508521Vz_obs (mg) / (ng / ml) 0.000336068Cl_obs (mg) / (ng / ml) / h 1.38798E-05Vss_obs (mg) / (ng / ml) 0.00042618

[0373] The data in Table 3 demonstrate average plasma concentration of Semaglutide at different time points. The pharmacokinetic profile are shown in Tables 4 and 5. Orally administered crystalline Semaglutide formulation with vitamin E showed almost 7 fold increase in AUC when compared to Rybelsus®. Semaglutide was analyzed in plasma using a specific enzyme linked immunosorbent assay (ELISA). Pharmacokinetic endpoints were determined using noncompartmental methods using PK Solutions software. AUC was approximated using the trapezoidal rule on the observed concentrations. C max for each Semaglutide dose was derived as the maximum of all valid concentrations, and t max was then determined as the corresponding time point to C max. The terminal elimination rate constant (X z) was estimated by log-linear regression on the terminal part of the concentration-time curve, and t * was then calculated as t * = In 2 / A Z.Example 13: A comparative studies between oral Semaglutide Tablets containing crystalline semaglutide with Vitamin E vs Tablets containing crystalline Semaglutide without Vitamin E Formulation in Guinea pigs

[0374] The present Example demonstrates that oral formulations of crystalline Semaglutide with Vitamin E exemplary of oral formulations disclosed herein havetherapeutically effective pharmacokinetic profiles, e.g., with advantageous properties as compared to oral administration of the crystalline semaglutide without vitamin E. The present example demonstrates a crystalline semaglutide Tablet formulation containing Vitamin E can be more effective than crystalline semaglutide formulation without vitamin E formulation demonstrating that the vitamin E is required for advantageous application of formulations of the present disclosure.

[0375] The present Example includes an oral formulation of crystalline peptide / protein embedded in Tablet prepared as follows: Commercially obtained semaglutide (synthetic semaglutide) from PRIVEEL PEPTIDES is a lyophilized white powder (>95% purity). The semaglutide was processed to prepare crystalline microparticles containing semaglutide according to the procedure mentioned under Example 1. The crystalline microparticle of semaglutide was then lyophilized after washing with cold isopropanol. The amount of lyophilized semaglutide was quantified against a reference standard using a C18 reverse phase HPLC column (gradient elution, solvent A (0.1% TFA in water), solvent B (0.1% TFA in acetonitrile)). The lyophilized semaglutide (dosage mentioned under Table 8) was then used to prepare tablets containing following composition: The tablets were then stored at 4°C until further use.Table 41. Tablet composition of Crystalline Semaglutide with Vitamin E

[0376] Each 3 mg of crystalline semaglutide used for making tablet without vitamin E contains following formulation as follows:Table 42. Tablet composition of Crystalline Semaglutide without Vitamin ETable 43. Experimental Design for Pharmacokinetics Analysis

[0377] Pharmacokinetic analysis was conducted in male Guinea pigs weighing 300 to 400 grams for 4 days. Experimental design is shown in Table 8. Animals were acclimatized for 5-7 days under laboratory conditions. On the day of experiment all the animals were fasted overnight prior to dosing (food but not water withheld). Following the fasting period, the animals were weighed and the test items were administered via oral route as per the groupings. In the oral route of administration, animals were dosed 1 tablet / animal and tablets were suspended in 0.5% CMC. Blood samples were collected at aforementioned time points 0 min, 15 min, Ihr, 2hrs, 6hrs, 24 hrs, 48 hrs, 72 hrs and 96 hrs into an EDTA-blood collection tubes for the separation of plasma to determine Semaglutide levels by ELISA. Plasma samples collected at different time points were analyzed by ELISA kit according to Krishgen Biosystems protocol. This kit is based on sandwich enzyme-linked immuno -sorbent assay technology. An antibody, anti-semaglutide was pre-coated onto a 96-well plate.Standards, test samples, and biotin conjugated reagent were added to the wells and incubated. The HRP-conjugated reagent was then added, and the whole plate was incubated. Unbound conjugates were removed using wash buffer at each stage. TMB substrate was used to quantify the HRP enzymatic reaction. After TMB substrate was added, only wells that contain sufficient Semaglutide will produce a blue colored product, which then changes toyellow after adding the acidic stop solution. The intensity of the yellow color is proportional to the Semaglutide amount bound on the plate. The Optical Density (OD) was measured spectrophotometrically at 450 nm in a microplate reader, from which the concentration of Semaglutide can be calculated.Table 44: Mean Semaglutide levels summary table (Guinea pigs) in ng / mlTable 45: Pharmacokinetic profile of Crystalline Semaglutide Tablet without vitamin E in Guinea pigsLambda_z 1 / h 0.009478012 tl / 2 h 73.13212382Tmax h 2Cmax ng / ml 291.7913333CO ng / ml 35.67633333Clast_obs / Cmax 0.497091529AUC O-t ng / ml*h 17080.87158AUC 0-inf_obs ng / ml*h 32384.39652AUC 0-t / 0- inf_obs 0.527441404AUMC 0- inf_obs ng / ml*hA2 3769632.412MRT 0-inf_obs h 116.4027377Vz_obs (mg) / (ng / ml) 0.009773906Cl_obs (mg) / (ng / ml) / h 9.26372E-05Vss_obs (mg) / (ng / ml) 0.010783224Table 46: Pharmacokinetic profile of Crystalline Semaglutide Tablet with vitamin E in Guinea pigsLambda_z 1 / h 0.0413005 tl / 2 h 16.78302167Tmax h 6Cmax ng / ml 4595.672333CO ng / ml 18.72666667Clast_obs / Cmax 0.037491141AUC O-t ng / ml*h 211970.0503AUC 0-inf_obs ng / ml*h 216141.84AUC 0-t / 0- inf_obs 0.980698833AUMC 0- inf_obs ng / ml*hA2 6636653.616MRT 0-inf_obs h 30.70508521Vz_obs (mg) / (ng / ml) 0.000336068Cl_obs (mg) / (ng / ml) / h 1.38798E-05Vss_obs (mg) / (ng / ml) 0.00042618

[0378] The data in Table 9 demonstrate average plasma concentration of Semaglutide at different time points. The full pharmacokinetic profile are shown in Tables 10 and 11. Orally administered crystalline Semaglutide formulation without vitamin E showed lower AUC when compared with one containing vitamin E formulation. Semaglutide was analyzed in plasma using a specific enzyme linked immunosorbent assay (ELISA). Pharmacokinetic endpoints were determined using noncompartmental methods using PK Solutions software. AUC was approximated using the trapezoidal rule on the observed concentrations. C max for each Semaglutide dose was derived as the maximum of all valid concentrations, and t max was then determined as the corresponding time point to C max. The terminal elimination rate constant (X z) was estimated by log-linear regression on the terminal part of the concentration-time curve, and t * was then calculated as t * = In 2 / A Z.Example 14: A comparative studies between oral Semaglutide Tablets containing crystalline semaglutide with Vitamin E vs Tablets containing commercial amorphous Semaglutide with Vitamin E Formulation in Guinea pigs

[0379] The present Example demonstrates that oral formulations of crystalline Semaglutide with Vitamin E exemplary of oral formulations disclosed herein have therapeutically effective pharmacokinetic profiles, e.g., with advantageous properties ascompared to oral administration of the commercially available amorphous semaglutide with vitamin E. The present example demonstrates a crystalline semaglutide Tablet formulation containing Vitamin E can be more effective than amorphous semaglutide formulation with vitamin E formulation demonstrating that crystalline semaglutide is required for advantageous application of formulations of the present disclosure.

[0380] The present Example includes an oral formulation of crystalline peptide / protein embedded in Tablet prepared as follows: Commercially obtained semaglutide (synthetic semaglutide) from PRIVEEL PEPTIDES is a lyophilized white powder (>95% purity). The semaglutide was processed to prepare crystalline microparticles containing semaglutide according to the procedure mentioned under Example 1. The crystalline microparticle of semaglutide was then lyophilized after washing with cold isopropanol. The amount of lyophilized semaglutide was quantified against a reference standard using a C18 reverse phase HPLC column (gradient elution, solvent A (0.1% TFA in water), solvent B (0.1% TFA in acetonitrile)). The lyophilized semaglutide (dosage mentioned under Table 14) was then used to prepare tablets containing following composition: The tablets were then stored at 4°C until further use.Table 47. Tablet composition of Crystalline Semaglutide with Vitamin E

[0381] Each 3 mg of commercially available amorphous semaglutide used for making tablet containing magnesium stearate, microcrystalline cellulose, povidone, vitamin E and 3 mg of amorphous semaglutide as follows.Table 48. Tablet composition of Amorphous Semaglutide with Vitamin ETable 49. Experimental Design for Pharmacokinetics Analysis

[0382] Pharmacokinetic analysis was conducted in male Guinea pigs weighing 300 to 400 grams for 4 days. Experimental design is shown in Table 14. Animals were acclimatized for 5-7 days under laboratory conditions. On the day of experiment all the animals were fasted overnight prior to dosing (food but not water withheld). Following the fasting period, the animals were weighed and the test items were administered via oral route as per thegroupings. In the oral route of administration, animals were dosed 1 tablet / animal and tablets were suspended in 0.5% CMC. Blood samples were collected at aforementioned time points 0 min, 15 min, Ihr, 2hrs, 6hrs, 24 hrs, 48 hrs, 72 hrs and 96 hrs into an EDTA-blood collection tubes for the separation of plasma to determine Semaglutide levels by ELISA. Plasma samples collected at different time points were analyzed by ELISA kit according to Krishgen Biosystems protocol. This kit is based on sandwich enzyme-linked immuno -sorbent assay technology. An antibody, anti-semaglutide was pre-coated onto a 96-well plate. Standards, test samples, and biotin conjugated reagent were added to the wells and incubated. The HRP-conjugated reagent was then added, and the whole plate was incubated. Unbound conjugates were removed using wash buffer at each stage. TMB substrate was used to quantify the HRP enzymatic reaction. After TMB substrate was added, only wells that contain sufficient Semaglutide will produce a blue colored product, which then changes to yellow after adding the acidic stop solution. The intensity of the yellow color is proportional to the Semaglutide amount bound on the plate. The Optical Density (OD) was measured spectrophotometrically at 450 nm in a microplate reader, from which the concentration of Semaglutide can be calculated.Table 50: Mean Semaglutide levels summary table (Guinea pigs) in ng / mlTable 51: Pharmacokinetic profile of amorpohous Semaglutide Tablet with vitamin E inGuinea pigsLambda.z 1 / h 0.023285093 tl / 2 h 29.76785082Tmax h 6Cmax ng / ml 591.589CO ng / ml 48.06166667Clast_obs / Cmax 0.12686116AUC 0-t ng / ml*h 24472.22867AUC 0-inf_obs ng / ml*h 27695.30645AUC 0-t / 0- inf_obs 0.883623682AUMC 0- inf_obs ng / ml*hA2 1190929.327MRT 0-inf_obs h 43.00112471Vz_obs (mg) / (ng / ml) 0.004651972Cl_obs (mg) / (ng / ml) / h 0.000108322Vss_obs (mg) / (ng / ml) 0.004657951Table 52: Pharmacokinetic profile of crystalline Semaglutide Tablet with vitamin E inGuinea pigsLambda_z 1 / h 0.0413005 tl / 2 h 16.78302167Tmax h 6Cmax ng / ml 4595.672333CO ng / ml 18.72666667Clast_obs / Cmax 0.037491141AUC 0-t ng / ml*h 211970.0503AUC 0-inf_obs ng / ml*h 216141.84AUC 0-t / 0- inf_obs 0.980698833AUMC 0- inf_obs ng / ml*hA2 6636653.616MRT 0-inf_obs h 30.70508521Vz_obs (mg) / (ng / ml) 0.000336068Cl_obs (mg) / (ng / ml) / h 1.38798E-05Vss_obs (mg) / (ng / ml) 0.00042618

[0383] The data in Table 15 demonstrate average plasma concentration of Semaglutide at different time points. The full pharmacokinetic profile are shown in Tables 16 and 17. Orally administered crystalline Semaglutide formulation showed 9 fold increase in AUC when compared to amorphous semaglutide formulation with vitamin E. Semaglutide was analyzed in plasma using a specific enzyme linked immunosorbent assay (ELISA).Pharmacokinetic endpoints were determined using noncompartmental methods using PK Solutions software. AUC was approximated using the trapezoidal rule on the observed concentrations. C max for each Semaglutide dose was derived as the maximum of all valid concentrations, and t max was then determined as the corresponding time point to C max. The terminal elimination rate constant (X z) was estimated by log-linear regression on the terminal part of the concentration-time curve, and t * was then calculated as t * = In 2 / A z.Example 15: Pharmacokinetic Study of Semaglutide Formulation Enclosed in Fast Dissolving Tablets or capsule with vitamin E in mini pigs

[0384] The present Example demonstrates that oral formulations of crystalline Semaglutide exemplary of oral formulations disclosed herein have therapeutically effective pharmacokinetic profiles, e.g., with advantageous properties as compared to oral administration of the commercially available Rybelsus® Tablets. The present example demonstrates either a crystalline semaglutide fast dissolving Tablet formulation or a capsule formulation with vitamin E can be more effective than Rybelsus® demonstrating that crystalline semaglutide either in fast dissolving tablet form for sublingual delivery or in capsule form with vitamin E is required for advantageous application of formulations of the present disclosure.

[0385] The present Example includes an oral formulation of crystalline peptide / protein embedded in Tablet prepared as follows: Commercially obtained semaglutide (synthetic semaglutide) from PRIVEEL PEPTIDES is a lyophilized white powder (>95% purity). The semaglutide was processed to prepare crystalline microparticles containing semaglutide according to the procedure mentioned under Example 1. The crystalline microparticle of semaglutide was then lyophilized after washing with cold isopropanol. The amount of lyophilized semaglutide was quantified against a reference standard using a C18 reverse phase HPLC column (gradient elution, solvent A (0.1% TFA in water), solvent B (0.1% TFA in acetonitrile)). The lyophilized semaglutide (dosage mentioned under Table 20) was then used to prepare tablets containing following composition: The tablets were then stored at 4°C until further use.Table 53. Tablet composition of Fast Dissolving crystalline Semaglutide TabletTable 54. Capsule composition of crystalline SemaglutideTable 55. Experimental Design for Pharmacokinetics Analysis

[0386] Pharmacokinetic analysis was conducted in female mini pigs weighing 11+1 kgs for 7 days. Experimental design is shown in Table 20. Animals were acclimatized for 5-7 days under laboratory conditions. On the day of experiment all the animals were fasted overnight prior to dosing (food but not water withheld). Following the fasting period, the animals were weighed and the test items were administered via SC or IV or oral route as per the groupings. In the oral route of administration, animals were dosed 1 tablet / animal and tablets were suspended in 0.5% CMC. Blood samples were collected at aforementioned time points 0 hr, 0.5 hr, 1 hr, 2 hrs, 4hrs, 6 hrs, 8 hrs, 12 hrs, 24 hrs, 48 hrs, 96 hrs, 120 hrs, 144 hrs and 168 hrs into an EDTA-blood collection tubes for the separation of plasma to determine Semaglutide levels by ELISA. Plasma samples collected at different time points were analyzed by ELISA kit according to Krishgen Biosystems protocol. This kit is based on sandwich enzyme-linked immuno- sorbent assay technology. An antibody, anti-semaglutide was pre-coated onto a 96-well plate. Standards, test samples, and biotin conjugated reagentwere added to the wells and incubated. The HRP-conjugated reagent was then added, and the whole plate was incubated. Unbound conjugates were removed using wash buffer at each stage. TMB substrate was used to quantify the HRP enzymatic reaction. After TMB substrate was added, only wells that contain sufficient Semaglutide will produce a blue colored product, which then changes to yellow after adding the acidic stop solution. The intensity of the yellow color is proportional to the Semaglutide amount bound on the plate. The Optical Density (OD) was measured spectrophotometrically at 450 nm in a microplate reader, from which the concentration of Semaglutide can be calculated.Table 56: Subcutaneous PK profile of Ozempic injectionTable 57: Full PK analysis of SC administration of Ozempic injectionTable 58: Intravenous PK profile of Ozempic injectionTable 59: Full PK analysis of intravenous administration of Ozempic injectionTable 60: Oral PK profile of Rybelsus tablet (amorphous semaglutide)Table 61: Sublingual PK profile of Crystalline semaglutide tabletTable 62: Full PK analysis oral administration of Rybelsus and Crystalline semaglutide tabletTable 63: Oral PK profile of Crystalline semaglutide capsuleTable 64: Full PK analysis oral administration of Crystalline semaglutide capsule

[0387] The data in Tables 21, 23, 25, 26 and 28 demonstrate average plasma concentration of Semaglutide at different time points. The full pharmacokinetic profile are shown in Tables 22, 24, 27 and 29. Orally administered crystalline Semaglutide formulation either through sublingual route or in capsule form orally showed better pharmacokinetic profile than Rybelsus®. Semaglutide was analyzed in plasma using a specific enzyme linked immunosorbent assay (ELISA). Pharmacokinetic endpoints were determined using noncompartmental methods using PK Solutions software. AUC was approximated using the trapezoidal rule on the observed concentrations. C max for each Semaglutide dose was derived as the maximum of all valid concentrations, and t max was then determined as the corresponding time point to C max. The terminal elimination rate constant (X z) was estimated by log-linear regression on the terminal part of the concentrationtime curve, and t * was then calculated as t * = In 2 / A z.Example 16: Preparation of Fast Dissolving Tablets or capsule containing sugars or albumin or amylase semaglutide

[0388] The present Example includes an oral formulation of crystalline peptide / protein embedded in Tablet is prepared as follows: Commercially available semaglutide (synthetic semaglutide) from PRIVEEL PEPTIDES is a lyophilized white powder (>95% purity). The semaglutide is processed to prepare crystalline microparticles containing semaglutide with sugar / sugars (such as Trehalose, mannitol, sorbitol, sucrose, mannose etc. either alone or in combination of various sugars) or with proteins (such as albumin or amylase) according to the procedure mentioned under Example 1. The crystalline microparticle of semaglutide containing sugar or protein is then lyophilized after washing with cold isopropanol. The amount of lyophilized semaglutide is quantified against a reference standard using a C18 reverse phase HPLC column (gradient elution, solvent A (0.1% TFA in water), solvent B (0.1% TFA in acetonitrile)). The lyophilized semaglutide containing either sugar or protein is then used to prepare tablets or capsules containing following composition: The tablets or capsules are then stored at 4°C until further use.Table 65. Tablet composition of Fast Dissolving crystalline Semaglutide TabletTable 66. Capsule composition of crystalline SemaglutideExample 17: Preparation of Tablets containing sugars or albumin or amylase - semaglutide

[0389] The present Example includes an oral formulation of crystalline peptide / protein embedded in Tablet is prepared as follows: Commercially obtained semaglutide (synthetic semaglutide) from PRIVEEL PEPTIDES is a lyophilized white powder (>95% purity). The semaglutide is processed to prepare crystalline microparticles containing semaglutide with sugar / sugars (such as Trehalose, mannitol, sorbitol, sucrose, mannose etc. either alone or in combination of various sugars) or with proteins (such as albumin or amylase) according to the procedure mentioned under Example 1. The crystalline microparticle of semaglutide containing sugar or protein is then lyophilized after washing with cold isopropanol. The amount of lyophilized semaglutide is quantified against a reference standard using a C18 reverse phase HPLC column (gradient elution, solvent A (0.1% TFA in water), solvent B (0.1% TFA in acetonitrile)). The lyophilized semaglutide containing either sugar or protein is then used to prepare tablets or capsules containing following composition: The tablets or capsules are then stored at 4°C until further use.Table 67. Tablet composition of Crystalline Semaglutide with Vitamin ETable 68. Tablet composition of Crystalline Semaglutide without Vitamin EExample 18: Crystallization of Amylase Carrier Protein Containing Semaglutide

[0390] The present Example demonstrates formation of crystallized amylase carrier protein embedded with semaglutide for use in an oral formulation as disclosed herein. The present Example includes the recognition that semaglutide is exemplary of peptide / proteins for which an oral formulation would be advantageous.

[0391] The present Example utilizes Semaglutide. To produce amylase carrier protein containing Semaglutide microparticles, Semaglutide in the lyophilized form is dissolved in water at a concentration of 80 mg / mL. Semaglutide aliquots are then mixed with amylase from Aspergillus oryzae (Sigma Chemical Company) solution to make amylase crystals containing semaglutide using the methods described below.

[0392] Crystallization method for amylase containing embedded Semaglutide: A 500 pL aliquot of amylase (-200 mg / mL), in water is mixed 500 pL of semaglutide solution (~ 80 mg / mL) with 1000 pL of reagent containing 30% (w / v) PEG 8000, 100 mM Sodium acetate / Acetic acid pH 4.5 and 200 mM Lithium sulfate, and is incubated at room temperature overnight. The final concentration of the semaglutide in solution is 20 mg / mL. This mixture is then mixed using a votex and is left at room temperature. Amylase microparticles containing semaglutide are obtained on the following day. About 90% of the input amylase is formed microparticles containing embedded semaglutide by this method.

[0393] In order to estimate the amount of semaglutide embedded in amylase crystals, the amylase crystals containing semaglutide are harvested by centrifugation of the sample at 4000 x g. The supernatant is discarded. The crystals are then washed twice with 1 mL of amylase crystallization reagent (30% (w / v) PEG 8000, 100 mM sodium acetate / acetic acid pH 4.5 and 200 mM Lithium sulfate). The washed crystals are then lyophilized after the final wash with cold isopropanol. The amount of semaglutide in the lyophilized sample is then quantified against a reference standard using a C18 reverse phase HPLC column (gradient elution, solvent A (0.1% TFA in water), solvent B (0.1% TFA in acetonitrile)). About 8 to 10% of the semaglutide is getting embedded / trapped inside the lyophilized amylase crystals.Example 19: Pharmacokinetic Study of Semaglutide Formulation either in powder form in sachet or in suspension in vitamin D3 in mini pigs

[0394] The present Example demonstrates that oral formulations of crystalline Semaglutide exemplary of oral formulations disclosed herein have therapeutically effective pharmacokinetic profiles, e.g., with advantageous properties as compared to oral administration of the commercially available Rybelsus® Tablets. The present example demonstrates either a crystalline semaglutide in powder formulation in sachet or a suspension formulation with vitamin D3 can be more effective than Rybelsus® demonstrating that crystalline semaglutide either in powder form sachet for sublingual delivery or in suspension form with vitamin D3 is required for advantageous application of formulations of the present disclosure.

[0395] The present Example includes an oral formulation of crystalline peptide / protein embedded in powder form in sachet prepared as follows: Commercially obtained semaglutide (synthetic semaglutide) from PRIVEEL PEPTIDES is a lyophilized white powder (>95% purity). The semaglutide was processed to prepare crystalline microparticles containing semaglutide according to the procedure mentioned under Example 1. The crystalline microparticle of semaglutide was then lyophilized after washing with cold isopropanol. The amount of lyophilized semaglutide was quantified against a reference standard using a C18 reverse phase HPLC column (gradient elution, solvent A (0.1% TFA in water), solvent B (0.1% TFA in acetonitrile)). The lyophilized semaglutide (dosage mentioned under Table 37) was then used to prepare powder form in sachet containing following composition: The powder forms in sachet were then stored at 4°C until further use.Table 69. Suspension formulation composition of crystalline Semaglutide powder in Vitamin D3 for oral deliveryTable 70. Sachet formulation composition of crystalline Semaglutide powder for sublingual delivery (Formulation 1)Table 71. Sachet formulation composition of crystalline Semaglutide powder for sublingual delivery(Formulation 2)Table 72. Experimental Design for Pharmacokinetics AnalysisExperimental design is shown in Table 37. Animals were acclimatized for 5-7 days under laboratory conditions. On the day of experiment all the animals were fasted overnight prior to dosing (food but not water withheld). Following the fasting period, the animals were weighed and the test items were administered via oral route as per the groupings. In the oral route of administration, animals were dosed 1 tablet (sachet) / animal and tablets were suspended in Vitamin D3. Blood samples were collected at aforementioned time points O hr, 0.33 hr, 1 hr, 2 hrs, 4hrs, 8 hrs, 12 hrs, 24 hrs, 48 hrs, and96 hrs into an EDTA-blood collection tubes for the separation of plasma to determine Semaglutide levels by ELISA. Plasma samples collected at different time points were analyzed either by LCMS (SPE-LC / MS Bioanalytical Quantification of the Bio therapeutic Peptide, Semaglutide From Plasma by Waters Corporation 2023) or by ELISA kit according to Krishgen Biosystems protocol. This kit is based on sandwich enzyme-linked immuno- sorbent assay technology. An antibody, anti- semaglutide was pre-coated onto a 96-well plate. Standards, test samples, and biotin conjugated reagent were added to the wells and incubated. The HRP-conjugated reagent was then added, and the whole plate was incubated. Unbound conjugates were removed using wash buffer at each stage. TMB substrate was used to quantify the HRP enzymatic reaction. After TMB substrate was added, only wells that contain sufficient Semaglutide will produce a blue colored product, which then changes to yellow after adding the acidic stop solution. The intensity of the yellow color is proportional to the Semaglutide amount bound on the plate. The Optical Density (OD) was measured spectrophotometrically at 450 nm in a microplate reader, from which the concentration of Semaglutide can be calculated.Table 73: Oral PK profile of Rybelsus tablet (amorphous semaglutide)Table 74: Oral PK profile of Crystalline semaglutide Suspension in Vitamin D3Table 75: Sublingual PK profile of Crystalline semaglutide tablet formulation 1.Table 76: Sublingual PK profile of Crystalline semaglutide tablet formulation 2.

[0397] The data in Tables 38, 39, 40, and 41 demonstrate average plasma concentration of Semaglutide at different time points. The AUC pharmacokinetic profile are shown in Tables 38, 39, 40, and 41. Orally administered crystalline Semaglutide formulation either through sublingual route or in oral suspension form in Vitamin D3 form orally showed better pharmacokinetic profile than Rybelsus®. Semaglutide was analyzed in plasma using a specific enzyme linked immunosorbent assay (ELISA). Pharmacokinetic endpoints were determined using noncompartmental methods using PK Solutions software. AUC was approximated using the trapezoidal rule on the observed concentrations. C max for each Semaglutide dose was derived as the maximum of all valid concentrations, and t max was then determined as the corresponding time point to C max. The terminal elimination rate constant (X z) was estimated by log-linear regression on the terminal part of the concentration-time curve, and t * was then calculated as t * = In 2 / A Z.Example 20: Pharmacokinetic Study of Semaglutide Formulation either in powder form in sachet or in suspension in vitamin D3 in mini pigs

[0398] The present Example demonstrates that oral formulations of crystalline Semaglutide exemplary of oral formulations disclosed herein have therapeutically effective pharmacokinetic profiles, e.g., with advantageous properties as compared to oral administration of the commercially available Rybelsus® Tablets. The present example demonstrates either a crystalline semaglutide in powder formulation in sachet or a suspension formulation with vitamin D3 can be more effective than Rybelsus® demonstrating that crystalline semaglutide either in powder form sachet for sublingual delivery or in suspension form with vitamin D3 is required for advantageous application of formulations of the present disclosure.

[0399] The present Example includes an oral formulation of crystalline peptide / protein embedded in powder form in sachet prepared as follows: Commercially obtained semaglutide (synthetic semaglutide) from PRIVEEL PEPTIDES is a lyophilized white powder (>95% purity). The semaglutide was processed to prepare crystalline microparticles containing semaglutide according to the procedure mentioned under Example 1. The crystalline microparticle of semaglutide was then lyophilized after washing with cold isopropanol. The amount of lyophilized semaglutide was quantified against a reference standard using a C18 reverse phase HPLC column (gradient elution, solvent A (0.1% TFA in water), solvent B (0.1% TFA in acetonitrile)). The lyophilized semaglutide (dosage mentioned under Table 46) was then used to prepare powder form in sachet containing following composition: The powder forms in sachet were then stored at 4°C until further use.Table 77. Tablet formulation composition of amorphous Rybelsus for oral deliveryTable 78. Suspension formulation composition of Rybelsus Semaglutide powder in Vitamin D3 for oral deliveryTable 79. Suspension formulation composition of crystalline Semaglutide powder for Oral delivery inVitamin D3Table 80. Sachet formulation composition of crystalline Semaglutide powder for sublingual deliveryTable 81. Experimental Design for Pharmacokinetics Analysis

[0400] Pharmacokinetic analysis was conducted in female mini pigs weighing 11+1 kgs for 4 days. Experimental design is shown in Table 46. Animals were acclimatized for 5-7 days under laboratory conditions. On the day of experiment all the animals were fasted overnight prior to dosing (food but not water withheld). Following the fasting period, the animals were weighed and the test items were administered via oral route as per the groupings. In the oral route of administration, animals were dosed 1 tablet (sachet) / animal and tablets were suspended in Vitamin D3. Blood samples were collected at aforementioned time points O hr, 0.33 hr, 1 hr, 2 hrs, 4hrs, 8 hrs, 12 hrs, 24 hrs, 48 hrs, and96 hrs into an EDTA-blood collection tubes for the separation of plasma to determine Semaglutide levels by ELISA. Plasma samples collected at different time points were analyzed either by LCMS (SPE-LC / MS Bioanalytical Quantification of the Bio therapeutic Peptide, Semaglutide From Plasma by Waters Corporation 2023) or by ELISA kit according to Krishgen Biosystems protocol. This kit is based on sandwich enzyme-linked immuno- sorbent assay technology. An antibody, anti- semaglutide was pre-coated onto a 96-well plate. Standards, test samples, and biotin conjugated reagent were added to the wells and incubated. The HRP-conjugated reagent was then added, and the wholeplate was incubated. Unbound conjugates were removed using wash buffer at each stage. TMB substrate was used to quantify the HRP enzymatic reaction. After TMB substrate was added, only wells that contain sufficient Semaglutide will produce a blue colored product, which then changes to yellow after adding the acidic stop solution. The intensity of the yellow color is proportional to the Semaglutide amount bound on the plate. The Optical Density (OD) was measured spectrophotometrically at 450 nm in a microplate reader, from which the concentration of Semaglutide can be calculated.Table 82: Oral PK profile of Rybelsus tablet (amorphous semaglutide)Table 83: Oral PK profile of Rybelsus tablet (amorphous semaglutide) in Vitamin D3 suspensionTable 84: Oral PK profile of powder sachet (crystalline semaglutide) in Vitamin D3 suspension.Table 85: Sublingual PK profile of Crystalline semaglutide tablet formulation.

[0401] The data in Tables 47, 48, 49, and 50 demonstrate average plasma concentration of Semaglutide at different time points. The AUC pharmacokinetic profile are shown in Tables 47, 48, 49, and 50. Orally administered crystalline Semaglutide formulation either through sublingual route or in oral suspension form in Vitamin D3 form orally showed better pharmacokinetic profile than Rybelsus®. Semaglutide was analyzed in plasma using a specific enzyme linked immunosorbent assay (ELISA). Pharmacokinetic endpoints were determined using noncompartmental methods using PK Solutions software. AUC was approximated using thetrapezoidal rule on the observed concentrations. C max for each Semaglutide dose was derived as the maximum of all valid concentrations, and t max was then determined as the corresponding time point to C max. The terminal elimination rate constant (X z) was estimated by log-linear regression on the terminal part of the concentration-time curve, and t * was then calculated as t * = In 2 / A Z.Example 21: Pharmacokinetic Study of Semaglutide Formulation either in powder form in sachet or in suspension in vitamin D3 in mini pigs

[0402] The present Example demonstrates that oral formulations of crystalline Semaglutide exemplary of oral formulations disclosed herein have therapeutically effective pharmacokinetic profiles, e.g., with advantageous properties as compared to oral administration of the commercially available Rybelsus® Tablets. The present example demonstrates either a crystalline semaglutide in powder formulation in sachet or a suspension formulation with vitamin D3 can be more effective than Rybelsus® demonstrating that crystalline semaglutide either in powder form sachet for sublingual delivery or in suspension form with vitamin D3 is required for advantageous application of formulations of the present disclosure.

[0403] The present Example includes an oral formulation of crystalline peptide / protein embedded in powder form in sachet prepared as follows: Commercially obtained semaglutide (synthetic semaglutide) from PRIVEEL PEPTIDES is a lyophilized white powder (>95% purity). The semaglutide was processed to prepare crystalline microparticles containing semaglutide according to the procedure mentioned under Example 1. The crystalline microparticle of semaglutide was then lyophilized after washing with cold isopropanol. The amount of lyophilized semaglutide was quantified against a reference standard using a C18 reverse phase HPLC column (gradient elution, solvent A (0.1% TFA in water), solvent B (0.1% TFA in acetonitrile)). The lyophilized semaglutide (dosage mentioned under Table 56) was then used to prepare powder form in sachet containing following composition: The powder forms in sachet were then stored at 4°C until further use.Table 86. Rybelsus Semaglutide Tablet for oral deliveryTable 87. Suspension formulation composition of Rybelsus Semaglutide powder in Vitamin D3 for oral deliveryTable 88. Suspension formulation composition of crystalline Semaglutide powder for Oral delivery in Vitamin D3 (Formulationl)Table 89. Suspension formulation composition of crystalline Semaglutide powder for Oral delivery inVitamin D3 (Formulation 2)Table 90. Sachet formulation composition of crystalline Semaglutide powder for sublingual deliveryTable 91. Experimental Design for Pharmacokinetics Analysis

[0404] Pharmacokinetic analysis was conducted in female mini pigs weighing 11+1 kgs for 4 days. Experimental design is shown in Table 56. Animals were acclimatized for 5-7 days under laboratory conditions. On the day of experiment all the animals were fasted overnight prior to dosing (food but not water withheld). Following the fasting period, the animals were weighed and the test items were administered via oral route as per the groupings. In the oral route of administration, animals were dosed 1 tablet (sachet) / animal and tablets were suspended in Vitamin D3. Blood samples were collected at aforementioned time points O hr, 0.33 hr, 1 hr, 2 hrs, 4hrs, 8 hrs, 12 hrs, 24 hrs, and 48 hrs into an EDTA-blood collection tubes for the separation of plasma to determine Semaglutide levels by ELISA. Plasma samples collected at different time points wereanalyzed either by LCMS (SPE-LC / MS Bioanalytical Quantification of the Bio therapeutic Peptide, Semaglutide From Plasma by Waters Corporation 2023) or by ELISA kit according to Krishgen Biosystems protocol. This kit is based on sandwich enzyme-linked immuno- sorbent assay technology. An antibody, anti- semaglutide was pre-coated onto a 96-well plate. Standards, test samples, and biotin conjugated reagent were added to the wells and incubated. The HRP-conjugated reagent was then added, and the whole plate was incubated. Unbound conjugates were removed using wash buffer at each stage. TMB substrate was used to quantify the HRP enzymatic reaction. After TMB substrate was added, only wells that contain sufficient Semaglutide will produce a blue colored product, which then changes to yellow after adding the acidic stop solution. The intensity of the yellow color is proportional to the Semaglutide amount bound on the plate. The Optical Density (OD) was measured spectrophotometrically at 450 nm in a microplate reader, from which the concentration of Semaglutide can be calculated.Table 92: Oral PK profile of Rybelsus tablet (amorphous semaglutide)Table 93: Oral PK profile of Rybelsus tablet (amorphous semaglutide) in Vitamin D3 suspensionTable 94: Oral PK profile of Crystalline formulation of semaglutide in Vitamin D3 suspension (Formulation 1)Table 95: Oral PK profile of powder sachet (crystalline semaglutide) in Vitamin D3 suspension (Formulation 2).Table 96: Sublingual PK profile of Crystalline semaglutide tablet formulation.

[0405] The data in Tables 57, 58, 59, 60 and 61 demonstrate average plasma concentration of Semaglutide at different time points. The AUC pharmacokinetic profile are shown in Tables 57, 58, 59, 60 and 61. Orally administered crystalline Semaglutide formulation either through sublingual route or in oral suspension form in Vitamin D3 form orally showed better pharmacokinetic profile than Rybelsus®. Semaglutide was analyzed in plasma using a specific enzyme linked immunosorbent assay (ELISA). Pharmacokinetic endpoints were determined using noncompartmental methods using PK Solutions software. AUC was approximated using the trapezoidal rule on the observed concentrations. C max for each Semaglutide dose was derived as the maximum of all valid concentrations, and t max was then determined as the corresponding time point to C max. The terminal elimination rate constant (A z) was estimated by log-linear regression on the terminal part of the concentration-time curve, and 116 was then calculated as 116 = In 2 / A z.OTHER EMBODIMENTS

[0406] While we have described a number of embodiments, it is apparent that our basic disclosure and examples may provide other embodiments that utilize or are encompassed by the compositions and methods described herein. Therefore, it will be appreciated that the scope of is to be defined by that which may be understood from the disclosure and the appended claims rather than by the specific embodiments that have been represented by way of example.

[0407] All references cited herein are hereby incorporated by reference.

Claims

CLAIMSWhat is claims is:

1. A tablet that comprises or delivers semaglutide, which tablet comprises:(i) semaglutide polypeptide; and(ii) a pharmaceutically acceptable carrier.

2. The tablet of claim 1 , wherein all of the semaglutide polypeptide is crystallized.

3. The tablet of claim 1, comprising one or more of crystalline semaglutide, microcrystalline cellulose, providone K90, Vitamin E, magnesium stearate, dextrate, and crospovidon, or any combinations thereof.

4. The tablet of claim 1, comprising crystalline semaglutide, microcrystalline cellulose, providone K90, Vitamin E, and magnesium stearate.

5. The tablet of claim 1, comprising crystalline semaglutide, microcrystalline cellulose, providone K90, and magnesium stearate.

6. The tablet of claim 1, comprising crystalline semaglutide, dextrate, and crospovidon.

7. The tablet of claim 1, comprsing crystalline semaglutide, microcrystalline cellulose, and Vitamin E.

8. The tablet of any one of the preceding claims, wherein the tablet comprises .5-25% w / w crystalline semaglutide.

9. The tablet of any one of the preceding claims, wherein the tablet comprises .60-95% w / w microcrystalline cellulose.

10. The tablet of any one of the preceding claims, wherein the tablet comprises .5-5% w / w providone K90.

11. The tablet of any one of the preceding claims, wherein the tablet comprises 1-6% w / w Vitamin E.

12. The tablet of any one of the preceding claims, wherein the tablet comprises .5-6% w / w magnesium stearate.

13. The tablet of any one of the preceding claims, wherein the tablet comprises 60-95% w / w dextrate.

14. The tablet of any one of the preceding claims, wherein the tablet comprises 3-13% w / w crospovidon.

15. The tablet of claim 1, wherein the tablet comprises one or more .5-18% w / w crystalline semaglutide, 70-95% w / w microcrystalline cellulose, .5-5% w / w providone K90, 1-6% w / w Vitamin E, 60-95% w / w dextrate, .5-6% w / w magnesium stearate, and 3-13% w / w crospovidon, or any combinations thereof.

16. The tablet of claim 1, wherein the tablet comprises .5-18% w / w crystalline semaglutide, 70-95% w / w microcrystalline cellulose, .5-5% w / w providone K90, 1-6% w / w Vitamin E, and .5-6% w / w magnesium stearate.

17. The tablet of claim 1, wherein the tablet comprises .5-18% w / w crystalline semaglutide, 70-95% w / w microcrystalline cellulose, .5-5% w / w providone K90, and .5-6% w / w magnesium stearate.

18. The tablet of claim 1, wherein the tablet comprises .5-18% w / w crystalline semaglutide, 60-95% w / w dextrate, and 3-13% w / w crospovidon.

19. The tablet of claim 1, wherein the tablet comprises .5-18% w / w crystalline semaglutide, 70-95% w / w microcrystalline cellulose, and .1-6% w / w Vitamin E.

20. The tablet of any one of the preceding claims, wherein the capsule formulation comprises a pharmaceutically acceptable carrier comprising a Vitamin E agent.

21. The tablet of any one of the preceding claims, wherein the semaglutide polypeptide composition is embedded in a carrier polypeptide composition.

22. A method of delivering a semaglutide polypeptide to the bloodstream of a subject, the method comprising orally administering to the subject a tablet of any one of the preceding claims.

23. The method of claim 22, wherein the semaglutide polypeptide is delivered to the bloodstream through mouth tissue, optionally wherein the polypeptide is delivered to the blood stream via sublingual administration.

24. A method of delivering a semaglutide polypeptide to mouth tissue of a subject, the method comprising orally administering to the subject a tablet of any one of the preceding claims.

25. A method of producing a tablet of any one of the preceding claims, the method comprising crystallizing the carrier polypeptide composition and the semaglutide polypeptide composition, wherein the semaglutide polypeptide is embedded in the carrier polypeptide during crystallization.

26. The tablet or method of any one of the preceding claims, wherein the tablet is or comprises a therapeutic polypeptide / peptide, optionally wherein the therapeutic polypeptide / peptide is or comprises:(i) semaglutide or fragment thereof; or(ii) an analog of (i), optionally wherein the polypeptide / peptide is modified by one or more of pegylation, acetylation, amidation, lipidation, methylation, phosphorylation,glycosylation, glycation, sulfation, mannosylation, nitrosylation, acylation, palmitoylation, prenylation, fatty acids, or a combination thereof.

27. The tablet or method of any one of the preceding claims, wherein the semaglutide polypeptide has a molecular weight between about 1 kDa and 6 kDa.

28. The tablet or method of any one of the preceding claims, wherein the semaglutide polypeptide comprises about 1 mg to about 200 mg of the tablet.

29. The tablet or method of any one of the preceding claims, wherein the carrier polypeptide composition is or comprises amylase.

30. The tablet or method of any one of the preceding claims, wherein the carrier polypeptide composition is or comprises albumin.

31. The tablet of any one of the preceding claims, wherein the carrier polypeptide composition or the semaglutide polypeptide composition comprises a crystallized polypeptide composition wherein the crystals have an average particle size of less than 25 microns, e.g., less than 20, 15, 10, 5, 4, 3, 2, 1, 0.5 or 0.1 microns, optionally wherein the crystals of polypeptide have an average particle size that is between 0.1 microns and 0.5, 1, 2, 3, 4, 5, 10, 15, 20, or 25 microns or wherein the crystals of polypeptide have an average particle size that is between 0.1 micron and 0.1, 0.5, 1, 2, 3, 4, 5, 10, 15, 20, or 25 microns.

32. The tablet of any one of the preceding claims , wherein the carrier polypeptide composition comprises an amorphous polypeptide composition wherein the particles of polypeptide have an average particle size of less than 25 microns, e.g., less than 20, 15, 10, 5, 4, 3, 2, 1, 0.5 or 0.1 microns, optionally wherein the particles of polypeptide have an average particle size that is between 0.1 microns and 0.5, 1, 2, 3, 4, 5, 10, 15, 20, or 25 microns or wherein the particles of polypeptide have an average particle size that is between 0.1 micron and 0.5, 1, 2, 3, 4, 5, 10, 15, 20, or 25 microns.

33. The tablet or method of any one of any one of the preceding claims, wherein the cocrystal of carrier polypeptide composition and semaglutide polypeptide composition comprises lyophilized polypeptide.

34. The tablet or method of any one of the preceding claims, wherein the co-crystal of carrier polypeptide composition and semaglutide polypeptide composition comprises microcrystals or nanocrystals of polypeptide.

35. The tablet or method of any one of the preceding claims, further comprising a coating or shell.

36. The tablet or method of claim 35, wherein the coating or shell is or comprises Vitamin E.

37. The tablet or method of claim 35, wherein the coating or shell is or comprises cane or palm sugar.

38. The tablet or method of any one of claims 35-37, wherein the shell is hollow.

39. The tablet or method of any one of claims 35-37, wherein the shell is hollow and the shell is fully filled with space of a core.

40. The tablet or method of any one of claims 35-37, wherein the shell is hollow and the shell is partially filled with space of a core.

41. The tablet or method of any one of the preceding claims, wherein the tablet is in a shape of sphere, cube, cone, cylinder, half sphere, torus, pyramid, triangular prism, hexagonal prism, cuboid, hexagonal pyramid, hallow cylinder, octahedron, diamond, star prism, hexagonal diamond, star pyramid, pentagonal prism, L shape prism, dodecahedron, tetrahedron, or icosahedron, or a modification thereof, or a combination thereof.

42. The tablet or method of any one of the preceding claims, wherein the tablet is in a shape of sphere, or a modification thereof.

43. The tablet or method of claim 42, wherein the sphere is hollow.

44. The tablet or method of any one of the preceding claims, wherein the tablet is formulated for delivery to the gut, optionally wherein the tablet is formulated for delivery to stomach and / or intestine.

45. The tablet or method of any one of the preceding claims, wherein the tablet is administered orally, optionally wherein the tablet is administered buccally or sublingually.

46. The tablet or method of claim 35, wherein the tablet further comprises an enteric coating.

47. The tablet or method of any one of the preceding claims, wherein the tablet further comprises one or more excipients or additives selected from the group consisting of aggregationreducing agents, sugars or sugar alcohols, polysaccharides, stabilizers, hyaluronidase, buffering agents, preservatives, carriers, antioxidants, chelating agents, natural or synthetic polymers, cryoprotectants, lyoprotectants, surfactants, bulking agents, acidifying agents, ingredients to reduce injection site discomfort, antifoaming agents, alkalizing agents, vehicles, aggregation inhibitors, solubilizing agents, permeation enhancers, muco bioadhesive agents, tonicity modifiers, and stabilizing agents and combinations thereof, optionally wherein the one or more excipients or additives are individually or cumulatively present at a concentration between 0.1 mM and about 1,000 mM, between about 0.1 mM and about 500 mM, between about 0.1 mM and about 200 mM, or between about 0.1 mM and about 100 mM.

48. The tablet or method of claim 47, wherein:(i) the aggregation-reducing agent(s)s are selected from the group consisting of nicotinic acid, caffeine citrate, caffeine nicotinate, caffeine, octyl- P-D-glucopyranoside, and n-dodecyl-P- D-maltoside and optionally in combination with one or more of arginine, tryptophan, histidine,proline, cysteine, methionine, P-alanine, Potassium Glutamate, Arginine Ethylester, lysine, aspartic acid, glutamic acid, glycine, DTPA (diethylenetriaminepentaacetic acid), EGTA(aminopolycarboxylic acid), EDTA (Ethylenediaminetetraacetic acid), hydroxy propyl beta (HP-Beta) cyclodextrins, hydroxy propyl gamma (HP-Gamma) cyclodextrins, sulfo-butyl ether (SBE) cyclodextrins, TMAO (trimethylamine N-oxide), trehalose, ethylene glycol, betaine, xylitol, sorbitol, 6-(N-(7-nitrobenz-2-oxa-l,3-diazol-4-yl)amino)hexanoic acid (NBD-X), methyl acetyl phosphate (MAP), citraconic anhydride, pyrophosphate, citrate, and combinations thereof;(ii) the tonicity modifier(s) are selected from the group consisting of arginine, cysteine, histidine, glycine, sodium chloride, potassium chloride, sodium citrate, saccharides such as sucrose, glucose, dextrose, glycerin or mannitol, and combinations thereof;(iii) the antioxidant(s) are selected from the group consisting of glycine, lysine, EDTA, DTPA, sorbitol, mannitol, ascorbic acid, ascorbyl palmitate, butylated hydroxy anisole, butylated hydroxytoluene, hypophosphorous acid, mono thioglycerol, propyl gallate, sodium bisulfite, sodium formaldehyde sulfoxylate, sodium metabisulfite, sodium thiosulfate, sufur dioxide, tocopherol, and combinations thereof;(iv) the lyoprotectant(s) are selected from the group consisting of sucrose, lactose, trehalose, dextran, erythritol, arabitol, xylitol, sorbitol, maltose, lactulose, maltulose, glucitol, maltitol, lactitol, isomaltulose and mannitol; amino acids, such as arginine or histidine or proline or glycine; lyotropic salts, such as magnesium sulfate; propylene glycol, glycerol, poly(ethylene glycol), or poly(propylene glycol); gelatin, dextrins, modified starch, carboxymethyl cellulose, and combinations thereof; or(v) the permeation enhancer(s) are selected from the group consisting of bile salts, e.g., tri-hydroxy salts sodium cholate, sodium glyco-cholate, sodium taurocholate and di-hydroxy salt, sodium deoxy cholate, sodium glyco-deoxy cholate, sodium tauro-deoxy cholate; fatty acids, their salt and esters, e.g., oleic acid, lauric acid, cod liver oil extract, sodium laurate, sodium caprate, glyceryl monostearate, di-ethylene glycol mono ethyl ether and various sucrose fatty acid esters, medium-chain fatty acid glycerides, polycaprolactoneomega-3 fatty acids, lecithin (phosphatidylcholine), lysophosphatidylcholine; surfactants, e.g., sodium dodecyl (lauryl) sulphate, poly sorbates (polysorbate 80), laureths, brijs and benzalkonium chloride; complexing agents, e.g., cyclodextrins, dextran sulphate, dextran sulphate, sodium edetate; co-solvents, e.g., ethanol and propylene glycol, combination of 1% oleic acid and 5% / 10% polyethylene glycol200, 2% glyceryl mono laurate and 40% alcohol, sodium caprate and alcohol or propylene glycol, 10% lauric acid in propylene glycol, polyoxyethylene, 2,3-lauryl ether, menthol, sodium caprate, sodium caprylate, sodium glycodeoxycholate, glycol; polysaccharides, e.g., chitosan and chitosan glutamate; and others such as, aprotinin, benzalkonium chloride, cetylpyridinium chloride, cetyltrimethyl ammonium bromide, sodium salicylate, lysophosphatidylcholine, methoxysalicylate, methyloleate , sodium edta, sulfoxides, various alkyl glycosides, ethylenediamide tetra acetic acid (edta), tartaric acid; lyotropic salts, such as magnesium sulfate; propylene glycol, glycerol, poly(ethylene glycol), or poly(propylene glycol); gelatin, dextrins, modified starch, carboxymethyl cellulose, and combinations thereof;(vi) the absorption enhancer(s) are selected from the group consisting of surfactants, cholesterol, glycerides, salicylates, bile salts, chelating agents, sodium caprate, a salt of capric acid and others include N-(5-chlorosalicylol)-8-aminocaprylic acid (5-CNAC), 4-((4-chloro-2- hydroxybenzoyl))-amino) butanoic acid (4-CNAB) and N-(8-(2-hydroxybenzoyl))-amino) caprylic acid, also known as salcaprozate sodium (SNAC, caprylic acid, C8, castor oil, medium chain, acyl carnitine, EDTA, glyceryl monolaurate, bovine P-casein, tocopherol succinate glycol chitosan conjugates, lecithins, glyceryl monostearate (GMS), chitosan and alginate, PLGA, silica, stearic acid, oleic acid, hydrogenated castor oil, and glyceryl trimyristate, etoposide phosphate (Vepesid®), sulindac (Clinoril®), enalapril maleate (Vasotec®), ramipril (Altace®), olmesartan medoxomil (Benicar®), valacyclovir (Valtrex®), midodrine (Amatine®), gabapentin enacarbil (Horizant®), sulfasalazine (Azulfidine®), and combinations thereof; or(vii) the muco bioadhesive agent(s) are selected from the group consisting of sucrose, lactose, trehalose, dextran, erythritol, arabitol, xylitol, sorbitol, maltose, lactulose, maltulose, glucitol, maltitol, lactitol, isomaltulose and mannitol; amino acids, such as arginine or histidine or proline or glycine; lyotropic salts, such as magnesium sulfate; propylene glycol, glycerol, poly(ethylene glycol), or poly(propylene glycol); gelatin, dextrins, modified starch, carboxymethyl cellulose, and combinations thereof. Mucoadhesive system such as from naturae, e.g., gelatin, agarose, chitosan, hyaluronic acid and synthetic polymers, e.g., polyvinylpyrolidone (PVP), polycrylates, polyvinyl alcohol, sodium carboxymethyl cellulose (SCMC) and pectin, all anionic-type polymers, chitosan (cationic type), and hydroxypropyl methylcellulose (HPMC) as a nonionic polymer, polyacrylic acid (PAA) derivatives (CP934, CP940, PCP), 15% CMC and 35% CP, copolymers of acrylic acid and poly(ethylene glycol) monomethylethermonomethacrylate (PEGMM), eudragitl NE40D as a neutral poly(ethylacrlate methylmethacrylate, hydrophilic polymers, e.g., methocel K4M, methocel K15M, SCMC 400, Cekol 700, Cekol 10000, CP934P, CP971P and CP974P, carboxyvinyl polymer and triethanolamine, HPC(hydroxy propyl celluose), CP (carbopol 934P), carbopol (CP) EX- 55 CMC (sodium carboxymethyl cellulose), HPMC (hydroxy propyl methyl cellulose), HEC (hydroxy ethyl cellulose), PIP [poly(isoprene)], PIB [poly (Isobutylene)], xanthum gum, locust bean gum, pectin, polycarbophil, benzyl esters, hydroxyethylcellulose, poly(acrylic acid), poly(acrylic acid-co-acrylamide), poly(acrylic acid-co-methyl methacrylate), poly(acrylic acid- co-butylacrylate), HEMA copolymerized with Polymeg® (polytetramethylene glycol), Cydot® (bioadhesive polymeric blend of CP and PIB), formulation consisting of PVP, cetylpyridinium chloride (as stabilizer), chitosan chloride, polyethylene oxide, polymethylvinylether / maleic anhydride (PME / MA), and tragacanth, poly ethyleneglycol monomethylether monomethacrylate, drum dried waxy maize starch (DDWM), carbopol 974P, and sodium stearylfumarate, and cellulose derivatives; hyderogels-acrylic acid (polar) and butyl acrylate (apolar), and combinations thereof.

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