Compositions and methods for enhancing the systemic bioavailability of orally administered polypeptide therapeutics

Abstract

The present invention provides methods for increasing the systemic bioavailability of a polypeptide therapeutic administered orally, via oral gavage, via nasogastric gavage, or intragastric administration, comprising administering the polypeptide therapeutic to the stomach of a human or non-human animal as part of one or more dosage forms together with an alginate oligomer and, optionally, a gastrointestinal permeation enhancer, wherein the one or more dosage forms do not, in addition to the polypeptide therapeutic, the alginate oligomer, and, if used, the gastrointestinal permeation enhancer, have a coating that provides substantial protection from the gastric environment. Such methods allow for the systemic treatment or prevention of diseases, conditions, or complications thereof that are responsive to or prevented by the polypeptide therapeutic via oral, oral gavage, via nasogastric gavage, or intragastric administration. Further provided are compositions comprising an alginate oligomer, a polypeptide therapeutic, and, optionally, a gastrointestinal permeation enhancer for use in such methods.

Description

[Technical Field]

[0001] The present invention relates to new and improved compositions and methods for enhancing the absorption of therapeutic polypeptides from the stomach. This, in turn, enhances the systemic bioavailability of therapeutic polypeptides administered by routes requiring passage through the stomach, such as orally, by oral gavage, nasogastric gavage, and intragastric administration. More specifically, the present invention relates to the use of alginate oligomers in combination with therapeutic polypeptides to enhance absorption from the stomach. The present invention is particularly useful in the context of oral administration of therapeutic polypeptides, which are typically administered by injection due to their low oral bioavailability. Increasing the systemic bioavailability of orally administered therapeutic polypeptides can enhance the effectiveness of therapeutic interventions and enable more convenient and comfortable treatments than parenteral approaches. [Background technology]

[0002] The efficacy of polypeptide therapeutics is often limited by their ability to reach their target. The mucosal surfaces of the lower gastrointestinal tract are attractive target sites for drug delivery due to their vascularity and large surface area. However, the primary route to these surfaces involves passage through the stomach, making polypeptide therapeutics particularly susceptible to the harsh chemical and enzymatic conditions of the stomach. The very low pH and high concentration of pepsin proteases cause significant degradation of polypeptides passing through the stomach. Furthermore, the entire gastrointestinal tract (mouth, pharynx, esophagus, stomach, small intestine, large intestine, and anus) is lined with a mucus layer that serves as a physical barrier between the epithelial cell layer and the GI (digestive) tract lumen. To enter the systemic circulation, therapeutic polypeptides must traverse the charged and complex polymeric "mesh" that makes up the mucus layer. The thickness of the mucus barrier is not uniform but varies, ranging from approximately 1 mm in the stomach to 100–150 μm in the colon and rectum.

[0003] Therefore, therapeutic polypeptides have traditionally been administered by injection, particularly intravenous or subcutaneous injection, to maximize systemic bioavailability. However, administration by injection is uncomfortable and inconvenient, and therefore, for polypeptide therapeutics that require regular administration to maintain therapeutically effective levels throughout the body, patient adherence to the treatment regimen becomes a significant challenge. The manufacturing and administration costs of injectable pharmaceuticals are often higher than those of oral pharmaceuticals, at least in part due to the need for sterility and hardware and the need for medically trained professionals to administer the drug. The solid waste associated with injectable drugs also creates biohazards and environmental burdens. Therefore, it would be more ideal if therapeutic polypeptides could be administered orally to patients.

[0004] Alginate is a natural polysaccharide that has found multiple uses in both clinical (e.g., in wound dressings, as an excipient in drug formulations, and in anti-heartburn preparations) and non-clinical (e.g., in food preparations) applications. Alginate is a linear polymer of (1-4)-linked β-D-mannuronic acid (M) and / or its C-5 epimer, α-L-glucuronic acid (G). The primary structure of alginate can vary greatly. M and G residues can be organized as homopolymer blocks of adjacent M or G residues, as blocks of alternating M and G residues, or with single M or G residues interposed between these block structures. Alginate molecules can contain some or all of these structures, and such structures may not be uniformly distributed throughout the polymer. At the extremes, homopolymers of glucuronic acid (polyglucuronic acid) or homopolymers of mannuronic acid (polymannuronic acid) exist.

[0005] Alginate has been isolated from marine brown algae (e.g., certain species of Durvillea, Lessonia, and Laminaria) and bacteria such as Pseudomonas aeruginosa and Azotobacter vinelandii. Other Pseudomonads (e.g., Pseudomonas fluorescens, Pseudomonas putida, and Pseudomonas mendocina) retain the genetic ability to produce alginate and, although they do not produce detectable levels of alginate in the wild, can be induced to do so.

[0006] Alginate is synthesized as a polymannuronic acid, and G residues are formed by the action of epimerases (specifically C-5 epimerases) on M residues in this polymer. In alginate extracted from algae, the G residues are primarily organized as G-blocks because the enzymes involved in alginate biosynthesis in the algae preferentially introduce G adjacent to another G, thus converting the chain of M residues into G-blocks.

[0007] Alginates are typically isolated from natural sources as large, high molecular weight polymers (e.g., average molecular weights ranging from 300,000 to 500,000 daltons). However, it is known that such large alginate polymers can be degraded or broken down, for example, by chemical or enzymatic hydrolysis, to produce lower molecular weight alginate structures. Commercially used alginates typically have average molecular weights ranging from 100,000 to 300,000 daltons (and such alginates are still considered large polymers), although alginates with average molecular weights of approximately 35,000 daltons have been used as excipients in pharmaceuticals.

[0008] Alginate oligomers have been recognized for their ability to alter the physical properties of isolated sputum (from patients with cystic fibrosis and normal controls) and mucus samples from the lung and gastrointestinal tract (Non-Patent Document 1, Non-Patent Document 2, Non-Patent Document 3, Non-Patent Document 4, Non-Patent Document 5, Non-Patent Document 6, Non-Patent Document 7).

[0009] This has led to previous proposals to exploit such properties to enhance drug delivery across mucosal surfaces, including macromolecular drugs (see U.S. Patent Nos. 5,629,997 and 5,729,997). There are no particular obstacles to implementing such proposals in the context of mucosal surfaces that are accessible without the need to expose the alginate oligomer and drug to gastric conditions, such as exposed mucus membranes, the respiratory tract, the genitourinary tract, the colon, the rectum, the anus, the mouth, the pharynx, or the esophagus. However, in the context of administration of drugs, particularly polypeptide therapeutics, by routes that require passage through the stomach, such as oral, orogastric, nasogastric, and intragastric routes, it has thus far appeared essential to enhance uptake of these therapeutic polypeptides and alginate oligomers by protecting both the polypeptide and alginate oligomer from the harsh environment of the gastric environment (e.g., by providing an enteric coating to these elements), thereby enabling movement through the stomach and delivery to the intestine, where enhanced uptake can occur (see U.S. Patent No. 5,729,997).

[0010] The prevailing theory is that alginate oligomers can enhance polypeptide delivery through the mucus barrier due to (i) their electrostatic interactions with the mucin glycan moiety and peptide backbone of the mucin polymer, and (ii) cation chelation. The resulting effects lead to an increase in mucus pore size, reduced viscosity, and a decreased ability to inhibit polypeptide passage. These properties are thought to depend on the alginate oligomer remaining functionally and structurally intact and not precipitating or gelling. Alginate precipitates when the pH is lower than its pKa (3.6-3.8). The pH of the human stomach is 1.5-3.5, so it was thought that alginate oligomers would precipitate in gastric conditions, thus losing their mucus-altering function. Indeed, high-molecular-weight alginate readily "precipitates" in the stomach, forming a hydrogel barrier that is used in the treatment of GERD (gastroesophageal reflux disease), where alginate forms a physical barrier to prevent acid reflux. [Prior art documents] [Patent documents]

[0011] [Patent Document 1] International Publication No. 2007 / 039754 [Patent Document 2] International Publication No. 2008 / 125828 [Patent Document 3] International Publication No. 2010 / 109180 [Non-patent literature]

[0012] [Non-Patent Document 1] Pritchard et al., Mucin structural interactions with an alginate oligomer mucolytic in cystic fibrosis sputum, Vibrational Spectroscopy, pp. 102932 (2019) [Non-patent document 2] Ermund, A. et al., OligoG CF-5 / 20 normalizes cystic fibrosis mucus by chelating calcium, Clinical and Experimental Pharmacology and Physiology, 44, 639-647 (2017) [Non-patent document 3] Vitko, M. et al., A novel guluronate oligomer improves intestinal transit and survival in cystic fibrosis mice, Journal of Cystic Fibrosis, 6, 745-751 (November 15, 2016) [Non-patent document 4] Pritchard, M.F. et al., A New Class of Safe Oligosaccharide Polymer Therapy To Modify the Mucus Barrier of Chronic Respiratory Disease, Molecular Pharmaceutics, Vol. 13, No. 3, pp. 863-72 (March 7, 2016) [Non-Patent Document 5] Nordgaerd et al., Alterations in mucus barrier function and matrix structure induced by guluronate oligomers, Biomacromolecules, Vol. 15, No. 6, pp. 2294-300 (June 9, 2014). [Non-patent document 6] Sletmoen, M. et al., Oligoguluronate induced competitive displacement of mucin-alginate interactions: relevance for mucolytic function, Soft Matter, No. 8, p. 8413 (2012) [Non-Patent Document 7] Nordgaerd, CT, Draget, KI, Oligosaccharides as modulators of rheology in complex mucus systems, Biomacromolecules, Vol. 12, No. 8, pp. 3084-3090 (August 8, 2011) Summary of the Invention [Means for solving the problem]

[0013] As already mentioned, from the perspective of therapeutic polypeptides, the very low pH and high concentrations of proteases such as pepsin in the stomach can cause significant degradation and / or denaturation, resulting in loss of function and reduced dosage.

[0014] It has now surprisingly been discovered that alginates having an average molecular weight of less than 15,000 daltons, e.g., 2-100 monomer residues, referred to herein as alginic acid oligomers, may be used in conjunction with (or in combination with) therapeutic polypeptides without substantial additional protection from the stomach environment to facilitate uptake of therapeutic polypeptides from the stomach when administered via routes requiring passage through the stomach, such as orally, via oral gavage, nasogastric gavage, and intragastric routes. This, in turn, may increase the systemic bioavailability of the polypeptide, e.g., to the extent that therapeutically effective levels in the systemic circulation can be reached and / or such levels can be reached more quickly and / or maximum circulating levels can be increased. In certain embodiments, this may allow less polypeptide therapeutic to be administered to achieve the required therapeutic efficacy, which in turn may minimize side effects and increase patient convenience and compliance. The need to administer less polypeptide and potentially fewer associated components in the dosage forms used further translates into manufacturing efficiencies and cost savings.

[0015] Furthermore, it has been found that the use of gastrointestinal permeation enhancers (more specifically, gastrointestinal epithelial barrier permeability enhancers) in conjunction with alginate oligomers and therapeutic peptides when administered via routes requiring passage through the stomach, such as oral, oral gavage, nasogastric gavage, and intragastric routes, further enhances uptake of those therapeutic polypeptides from the stomach, without substantial additional protection from the stomach environment. This, in turn, may increase the systemic bioavailability of the polypeptide, for example, to the extent that it can reach therapeutically effective levels in the systemic circulation and / or reach such levels more quickly and / or increase maximum circulating levels. In certain embodiments, this may allow less polypeptide therapeutic to be administered to achieve the required therapeutic efficacy, which in turn may minimize side effects and increase patient convenience and retention. The need to administer less polypeptide and potentially fewer associated components in a dosage form further translates into manufacturing efficiencies and cost savings.

[0016] Thus, the present invention provides a method for increasing the systemic bioavailability of an orally, orogastrically, orally, orally, orally, orally, orally, orally, orally, orally, orally, orally, orally, administered polypeptide therapeutic agent, said method comprising administering said polypeptide therapeutic agent together with an alginate oligomer, and optionally a gastrointestinal permeation enhancer, to the stomach of a human or non-human subject as part of one or more dosage forms, wherein said one or more dosage forms, in addition to the polypeptide therapeutic agent, alginate oligomer, and, if used, the gastrointestinal permeation enhancer, do not have a coating that provides substantial protection from the gastric environment. [Brief explanation of the drawings]

[0017] [Figure 1] Representative DOSY spectra of tirzepatide (Tz) alone or in the presence of SNAC or C10 are shown. The top three traces are, from top to bottom, tirzepatide (light gray), tirzepatide + SNAC (black), and tirzepatide + C10 (dark gray). [Figure 2]Figure 1 shows the time course of diffusion of semaglutide through artificial mucus in a Transwell device in the presence of SNAC and / or Oligo G at 37°C. Bars from left to right: 15, 30, 45 and 240 minutes. [Figure 3] Diffusion of tirzepatide (TZ) through artificial mucus in a Transwell device after 60 minutes at 37° C. in the presence of alone or in combination with C10, SNAC and / or Oligo G (OG). [Figure 4] Figure 1 shows the diffusion of liraglutide (LZ) through artificial mucus in a Transwell device after 60 minutes at 37°C in the presence of SNAC and / or Oligo G (OG) alone or in combination. [Figure 5] Figure 1 shows the diffusion of semaglutide (SG) through artificial mucus in a Transwell device after 60 minutes at 37°C in the presence of SNAC and / or Oligo G (OG) alone or in combination. [Figure 6] Plasma concentrations of semaglutide in rats are shown after oral gavage of semaglutide alone (solid line, semaglutide (6.67 mg / kg)), or semaglutide with SNAC (long-dashed line, semaglutide (6.67 mg / kg) + SNAC (400 mg / kg)), or semaglutide with SNAC and Oligo G (short-dashed line, semaglutide (6.67 mg / kg) + SNAC (400 mg / kg) + Oligo G (333 mg / kg)). All doses are based on an assumed weight of 300 grams per animal. [Figure 7] Plasma concentrations of tirzepatide in rats after oral gavage of tirzepatide alone (solid line, tirzepatide (7.65 mg / kg)), tirzepatide with C10 (long dashed line, tirzepatide (7.65 mg / kg) + C10 (257.87 mg / kg)), or tirzepatide with C10 and oligo G (short dashed line, tirzepatide (7.65 mg / kg) + C10 (257.87 mg / kg) + oligo G (333 mg / kg)). All doses are based on an assumed weight of 300 grams per animal. [Figure 8]Plasma concentrations of semaglutide in rats after oral gavage of semaglutide (6.67 mg / kg) with SNAC (400 mg / kg) (solid line), semaglutide (6.67 mg / kg) with SNAC (180 mg / kg) and Oligo G (100 mg / kg) (short-dashed line), semaglutide (6.67 mg / kg) with SNAC (400 mg / kg) and Oligo G (100 mg / kg) (long-dashed line), and semaglutide (6.67 mg / kg) with SNAC (400 mg / kg) and Oligo G (333 mg / kg) (dotted line). All doses are based on an assumed weight of 300 grams per animal. [Figure 9] Figure 1 shows the systemic bioavailability of semaglutide within 90 minutes (calculated as the area under the curve) following oral administration of semaglutide with various combinations and levels of SNAC and oligoG: semaglutide 6.67 mg / kg, high-dose oligoG 333 mg / kg, low-dose oligoG 100 mg / kg, low-dose SNAC 180 mg / kg, high-dose SNAC 400 mg / kg. All doses are based on an assumed weight of 300 grams per animal. [Figure 10] Plasma concentrations of semaglutide in rats at 15 and 30 minutes after oral gavage of semaglutide (6.67 mg / kg) with SNAC (400 mg / kg) (grey bars) and semaglutide (6.67 mg / kg) with SNAC (400 mg / kg) and Oligo G (333 mg / kg) (black lines) are shown. All doses are based on an assumed weight of 300 grams per animal. [Figure 11] Plasma concentrations of tirzepatide in rats after oral gavage of tirzepatide (7.65 mg / kg) with C10 (128.9 mg / kg) (solid line) and tirzepatide (7.65 mg / kg) with C10 (192 mg / kg) and Oligo G (333 mg / kg) (dotted line). All doses are based on an assumed weight of 300 grams per animal. [Figure 12]Figure 1 shows the systemic bioavailability of tirzepatide within 90 minutes of oral administration of tirzepatide (7.65 mg / kg) with C10 (128.9 mg / kg) (solid line) and tirzepatide (7.65 mg / kg) with C10 (192 mg / kg) and Oligo G (333 mg / kg). All doses are based on an assumed weight of 300 grams per animal. [Figure 13] Plasma concentrations of tirzepatide in rats at 15 and 30 minutes after oral gavage of tirzepatide (7.65 mg / kg) with C10 (128.9 mg / kg) (gray bars) and tirzepatide (7.65 mg / kg) with C10 (192 mg / kg) and Oligo G (333 mg / kg) (black lines) are shown. All doses are based on an assumed weight of 300 grams per animal. [Figure 14] ) Systemic bioavailability of retatortide within 90 minutes following oral gavage of retatortide (6.67 mg / kg) with C10 (125 mg / kg) (gray line) and retatortide (6.67 mg / kg) with C10 (125 mg / kg) and Oligo G (250 mg / kg) (black line). All doses are based on an assumed weight of 300 grams per animal. [Figure 15] Figure 1 shows the plasma concentration of retatortide in rats 10 minutes after oral gavage of retatortide (6.67 mg / kg) with C10 (125 mg / kg) (gray bars) and retatortide (6.67 mg / kg) with C10 (125 mg / kg) and Oligo G (250 mg / kg) (black bars). All doses are based on an assumed weight of 300 grams per animal. [Figure 16] The chemical structures of liraglutide (A), semaglutide (B), tirzepatide (C), and retatortide (D) are shown. DETAILED DESCRIPTION OF THE INVENTION

[0018] The present invention further provides an alginate oligomer for use in a method for increasing the systemic bioavailability of an orally, orally gavaged, nasogastric, or intragastrically administered polypeptide therapeutic agent, the method comprising administering the polypeptide therapeutic agent together with the alginate oligomer, and optionally a gastrointestinal permeation enhancer, to the stomach of a human or non-human animal as part of one or more dosage forms, wherein the one or more dosage forms, in addition to the polypeptide therapeutic agent, alginate oligomer, and, if used, the gastrointestinal permeation enhancer, do not have a coating that provides substantial protection from the stomach environment.

[0019] Generally, the term "systemic bioavailability" as used herein refers to the proportion of an administered dose of an active pharmaceutical ingredient (API), e.g., a therapeutic polypeptide as defined herein, that reaches the systemic circulation in an active or reversibly reduced form. By definition, when an API is administered intravenously, its bioavailability is 100%. However, when it is administered by other routes, e.g., orally or subcutaneously, its bioavailability decreases (due to incomplete absorption and first-pass metabolism). Absolute systemic bioavailability is calculated as the relative exposure of an API in the systemic circulation after non-intravenous administration (e.g., oral or mucosal, e.g., inhalation), and is evaluated as the area under the plasma concentration versus time curve (AUC) compared to the API exposure after intravenous administration.

[0020] According to the present invention, the use of an alginate oligomer in combination with a therapeutic polypeptide, and optionally a gastrointestinal permeation enhancer, administered orally, by oral gavage, nasogastric gavage or intragastricly to the stomach of a human or non-human animal as part of one or more dosage forms results in a greater measure of absolute systemic bioavailability compared to administration of the therapeutic polypeptide, and optionally a gastrointestinal permeation enhancer, in an otherwise identical situation but without the alginate oligomer present.

[0021] AUC may be calculated as the definite integral of the concentration of drug in plasma as a function of time. Conveniently, this may be calculated using the trapezoidal rule.

[0022] In other words, the present invention provides a method for gastric uptake of an active or reversibly reduced-activity polypeptide therapeutic agent, said method comprising administering said polypeptide therapeutic agent together with an alginate oligomer, and optionally a gastrointestinal permeation enhancer, to the stomach of a human subject or non-human animal subject as part of one or more dosage forms, wherein said one or more dosage forms, in addition to the polypeptide therapeutic agent, alginate oligomer, and, if used, the gastrointestinal permeation enhancer, do not have a coating that provides substantial protection from the gastric environment.

[0023] Similarly, the present invention further provides an alginate oligomer for use in a method for gastric uptake of an active or reversibly reduced-activity polypeptide therapeutic agent, the method comprising administering the polypeptide therapeutic agent together with the alginate oligomer, and optionally a gastrointestinal permeation enhancer, to the stomach of a human subject or non-human animal subject as part of one or more dosage forms, wherein the one or more dosage forms, in addition to the polypeptide therapeutic agent, alginate oligomer, and, if used, the gastrointestinal permeation enhancer, do not have a coating that provides substantial protection from the gastric environment.

[0024] In other words, the present invention provides a method for absorbing an active or reversibly reduced-activity polypeptide therapeutic agent from the stomach of a human subject or non-human animal subject, said method comprising administering said polypeptide therapeutic agent together with an alginate oligomer, and optionally a gastrointestinal permeation enhancer, to the stomach of the human subject or non-human animal subject as part of one or more dosage forms, wherein said one or more dosage forms, in addition to the polypeptide therapeutic agent, alginate oligomer, and, if used, the gastrointestinal permeation enhancer, do not have a coating that provides substantial protection from the gastric environment.

[0025] Similarly, the present invention provides an alginate oligomer for use in a method for absorbing an active or reversibly reduced activity polypeptide therapeutic agent from the stomach of a human subject or non-human animal subject, the method comprising administering the polypeptide therapeutic agent together with the alginate oligomer, and optionally a gastrointestinal permeation enhancer, to the stomach of the human subject or non-human animal subject as part of one or more dosage forms, wherein the one or more dosage forms, in addition to the polypeptide therapeutic agent, alginate oligomer, and, if used, the gastrointestinal permeation enhancer, do not have a coating that provides substantial protection from the gastric environment.

[0026] In other words, the present invention provides a method for increasing the absorption of an active or reversibly reduced-activity polypeptide therapeutic agent from the stomach of a human subject or non-human animal subject, said method comprising administering said polypeptide therapeutic agent together with an alginate oligomer, and optionally a gastrointestinal permeation enhancer, to the stomach of the human subject or non-human animal subject as part of one or more dosage forms, wherein said one or more dosage forms, in addition to the polypeptide therapeutic agent, alginate oligomer, and, if used, the gastrointestinal permeation enhancer, do not have a coating that provides substantial protection from the gastric environment.

[0027] Similarly, the present invention further provides an alginate oligomer for use in a method for increasing absorption of an active or reversibly reduced-activity polypeptide therapeutic agent from the stomach of a human or non-human subject, the method comprising administering the polypeptide therapeutic agent together with the alginate oligomer, and optionally a gastrointestinal permeation enhancer, to the stomach of the human or non-human subject as part of one or more dosage forms, wherein the one or more dosage forms, in addition to the polypeptide therapeutic agent, alginate oligomer, and, if used, the gastrointestinal permeation enhancer, do not have a coating that provides substantial protection from the gastric environment.

[0028] Increased absorption from the stomach can be seen as an increase in systemic bioavailability over time, with a significant proportion of the target drug / dosage form remaining (retained) in the stomach upon administration. It may also be seen as a greater Cmax after entry into the stomach. It may also be seen as a more rapid rise in plasma concentration and / or a reduced lag in the time it takes for the plasma concentration of the therapeutic polypeptide to begin to rise. These observations are in comparison to administration of a therapeutic polypeptide, and optionally a gastrointestinal permeation enhancer, under otherwise identical circumstances, without the alginate oligomer.

[0029] In other words, the present invention provides a method for improving the effectiveness of an orally, orally, orally, orally, orally, orally, orally, or intragastrically, administered polypeptide therapeutic in the internal treatment or prevention of a disease or condition responsive to or prevented by a polypeptide therapeutic, or a complication thereof, said method comprising administering said polypeptide therapeutic together with an alginate oligomer, and optionally a gastrointestinal permeation enhancer, to the stomach of a human subject or non-human animal subject as part of one or more dosage forms, wherein said one or more dosage forms, in addition to the polypeptide therapeutic, the alginate oligomer, and, if used, the gastrointestinal permeation enhancer, do not have a coating that provides substantial protection from the gastric environment.

[0030] Similarly, the present invention further provides an alginate oligomer for use in a method for improving the effectiveness of an orally, orally, orally, orally, orally, orally, or intragastrically, administered polypeptide therapeutic in the internal treatment or prevention of a disease or condition responsive to or prevented by a polypeptide therapeutic, or a complication thereof, said method comprising administering said polypeptide therapeutic together with said alginate oligomer, and optionally a gastrointestinal permeation enhancer, to the stomach of a human or non-human subject as part of one or more dosage forms, wherein said one or more dosage forms, in addition to the polypeptide therapeutic, alginate oligomer, and, if used, the gastrointestinal permeation enhancer, do not have a coating that provides substantial protection from the gastric environment.

[0031] "Improving the efficacy of a polypeptide therapeutic" refers to any positive effect on the therapeutic use of the polypeptide in question. This may include greater potency or potency at its site of action or pharmacological receptor, longer duration of pharmacological effect, fewer or milder side effects (systemic or local), a wider therapeutic window, greater circulating plasma levels, prolonged circulating plasma levels, higher Cmax, faster uptake, etc. These observations are compared to the administration of a therapeutic polypeptide in the absence of alginate oligomer and, optionally, a gastrointestinal permeation enhancer under otherwise identical circumstances. Improved efficacy may be seen in the requirement that less polypeptide be administered to achieve the same therapeutic result in essentially the same manner.

[0032] In other words, the present invention provides a method for the systemic treatment or prevention of a disease or condition or a complication thereof that is responsive to or can be prevented by a polypeptide therapeutic, said method comprising administering said polypeptide therapeutic together with an alginate oligomer, and optionally a gastrointestinal permeation enhancer, as part of one or more dosage forms to the stomach of a human or non-human subject having, suspected of having, or at risk of having said disease or condition or a complication thereof, said one or more dosage forms not bearing, in addition to the polypeptide therapeutic, the alginate oligomer, and, if used, the gastrointestinal permeation enhancer, a coating that provides substantial protection from the gastric environment.

[0033] Similarly, the present invention further provides an alginate oligomer for use in a method for the systemic treatment or prevention of a disease or condition or a complication thereof that is responsive to or can be prevented by a polypeptide therapeutic, said method comprising administering said polypeptide therapeutic together with said alginate oligomer, and optionally a gastrointestinal permeation enhancer, to the stomach of a human or non-human subject having, suspected of having, or at risk of having said disease or condition or complication as part of one or more dosage forms, wherein said one or more dosage forms, in addition to the polypeptide therapeutic, alginate oligomer, and, if used, the gastrointestinal permeation enhancer, do not have a coating that provides substantial protection from the gastric environment.

[0034] According to the present invention, systemic treatment or prevention involves administration of a therapeutic polypeptide in a manner that results in the polypeptide entering the blood circulation and distributing to areas of the subject's body that are spatially distant from the site of administration, specifically the stomach. Preferably, circulating levels of the polypeptide are above the level required to achieve a therapeutic effect. Systemic treatment includes treatment of systemic conditions, conditions with lesions in several spatially distributed locations, and / or conditions with a single, localized lesion distant from the site of administration.

[0035] The methods of the invention may include, following administration of a polypeptide therapeutic together with an alginate oligomer and, optionally, a gastrointestinal permeation enhancer, measuring the amount or concentration of the polypeptide therapeutic in the subject's plasma, e.g., within 360, 330, 300, 270, 240, 210, 180, 120, 60, 30, 20, 15, 10, or 5 minutes after administration of the last administered polypeptide therapeutic, alginate oligomer, or gastrointestinal permeation enhancer. In other embodiments, the methods of the invention may include, or may further include, in addition to the measuring steps described above, a step in which the systemic bioavailability or absolute systemic bioavailability of the polypeptide therapeutic is calculated, e.g., within 360, 330, 300, 270, 240, 210, 180, 120, 60, 30, 20, 15, 10, or 5 minutes after administration of the last administered polypeptide therapeutic, alginate oligomer, or gastrointestinal permeation enhancer.

[0036] Further contemplated is the use of alginate oligomers in the manufacture of medicaments for use in the methods described above, which may be in one or more of the dosage forms described above for administration to a human or non-human animal subject during such methods.

[0037] By "substantial protection from the gastric environment" is meant that a substantial proportion of the targeted agent administered to the stomach, i.e., the agent under consideration (e.g., polypeptide therapeutic agent, alginate oligomer and / or gastrointestinal permeation enhancer), is not exposed to the gastric environment, or alternatively, a substantial proportion of the agent or dosage form of which the agent is a part remains in the stomach to which it is administered, while potentially deleterious effects of the gastric environment on the targeted agent are prevented, inhibited or mitigated.

[0038] Thus, "substantial protection from the gastric environment" may be considered to be a means, such as a coating as described above, that delays exposure of the polypeptide therapeutic agent, alginate oligomer and / or gastrointestinal permeation enhancer to the gastric environment or delays the potentially deleterious effects of the gastric environment on the targeted agent for a time period approaching or greater than the time that a significant proportion of the agent or dosage form of which the agent is a part remains in the stomach to which it is administered.

[0039] Alternatively, this may be expressed as meaning that a significant proportion of the agent or dosage form of which the agent is a part remains in the stomach to which it is administered, that only a small proportion of the targeted agent (e.g., polypeptide therapeutic agent, alginate oligomer and / or gastrointestinal permeation enhancer) administered to the stomach is exposed to the gastric environment, or that the potentially deleterious effects of the gastric environment on the targeted agent are limited to a small proportion of the agent.

[0040] In more specific terms, "substantial protection from the gastric environment" may be expressed as meaning that a substantial proportion of the targeted agent (e.g., polypeptide therapeutic agent, alginate oligomer and / or gastrointestinal permeation enhancer) administered to the stomach, or a substantial proportion of the dosage form of which the targeted agent is a part, is stabilized (becomes stable) in the gastric environment for the time that the agent, or a substantial proportion of the dosage form of which the agent is a part, remains in the stomach to which it is administered.

[0041] Alternatively, this may be expressed to mean that while a substantial proportion of the drug or dosage form of which the drug is a part remains in the stomach to which it is administered, a small proportion of the targeted drug (e.g., polypeptide therapeutic, alginate oligomer and / or gastrointestinal permeation enhancer) or dosage form of which the targeted drug is a part that is administered to the stomach is destabilized in the gastric environment (e.g., in the case of a drug, it is denatured, deteriorated, precipitated, solidified or inactivated / reduced in activity for any reason, and in the case of a dosage form, it is dissolved, dispersed or disintegrated).

[0042] Thus, in accordance with the present invention, the polypeptide therapeutic agent, alginate oligomer and / or gastrointestinal permeation enhancer are administered in one or more dosage forms that do not have a structure that may provide substantial protection from the gastric environment to the polypeptide therapeutic agent, alginate oligomer and / or gastrointestinal permeation enhancer of which they are a part.

[0043] A coating or any other dosage form structure that may provide "substantial protection from the gastric environment" may be considered to be a dosage form structure that combats, opposes, neutralizes, inhibits, or reduces the potentially harmful effects of the gastric environment on a targeted agent (e.g., a polypeptide therapeutic agent, an alginate oligomer, and / or a gastrointestinal permeation enhancer) or a dosage form of which the targeted agent is a part, particularly while a substantial proportion of the agent or dosage form of which the agent is a part remains in the stomach to which it is administered.

[0044] A "substantial proportion of the target drug" may be considered to be at least 70%, e.g., at least 75%, 80%, 85%, 90%, or 95% of the administered dose, measured by weight. A "substantial proportion of the dosage form" may be considered to be at least 70%, e.g., at least 75%, 80%, 85%, 90%, or 95% of the administered dosage form, measured by weight.

[0045] A "minor percentage of the target drug" may be considered to be 25% or less of the administered dose, e.g., 20%, 15%, 10%, or 5% or less, measured by weight. A "minor percentage of the dosage form" may be considered to be 25% or less of the administered dose, e.g., 20%, 15%, 10%, or 5% or less, measured by weight.

[0046] The time that a significant proportion of the target drug / dosage form remains (retains) in the stomach after administration varies between subjects, the drug in question, the components and properties of the dosage form of which the drug is a part, and any exogenous liquids or solids consumed by the dosage form, but can range from about 15 minutes to approximately 240 minutes after entry into the stomach, e.g., from about 15 minutes to about 20, 30, 40, 50, 60, 70, 80, 90, 100, 110, 120, 130, 140, 150, 160, 170, 180, 190, 200, 210, 220, 230, or 240 minutes, or from about 15, 20, 30, 40, 50, 60, 70, 80, 90, 100, 110, 120, 130, 140, 150, 160, 170, 180, 190, 200, 210, 220, or 230 minutes to about 240 minutes. Any range that may be comprised of the above values ​​is expressly contemplated.

[0047] This residence / retention time may be viewed as the time it takes for a drug, or a significant proportion of the dosage form of which the drug is a part, to pass through the stomach into which it has been administered and enter the intestine essentially unchanged (i.e., without exhibiting the deleterious effects of the gastric environment described above).

[0048] Thus, "substantial protection from the gastric environment" may be considered to be prolonged protection, i.e., protection for the polypeptide therapeutic agent, alginate oligomer and / or gastrointestinal permeation enhancer, or a substantial proportion of the dosage form of which it is a part, against exposure to the gastric environment or the potentially deleterious effects of the gastric environment on the target agent that lasts for a sufficient period of time to allow the agent, or a substantial proportion of the dosage form of which it is a part, to pass through the stomach to which it is administered and enter the intestine essentially unchanged.

[0049] Thus, the characteristic of "substantial protection from the gastric environment" may be considered to be met if the targeted agent (e.g., polypeptide therapeutic agent, alginate oligomer and / or gastrointestinal permeation enhancer), or a substantial proportion of the dosage form of which the targeted agent is a part, enters the intestine unchanged within about 240 minutes of entry into the stomach, e.g., within about 220, 200, 180, 160, 140, 120, 100, 80, or 60 minutes of entry into the stomach.

[0050] The term "gastric environment" refers to the physical and chemical conditions in the stomach lumen to which the polypeptide therapeutic agent, alginate oligomer, and / or gastrointestinal permeation enhancer are administered in accordance with the present invention. The exact conditions will vary between subjects, but typically have a pH of about 1.0 to about 3.5, e.g., about 1.2 to about 1.4, 1.6, 1.8, 2.0, 2.2, 2.4, 2.6, 2.8, 3.0, 3.2, 3.4, or 3.5, or about 1.4, 1.6, 1.8, 2.0, 2.2, 2.4, 2.6, 2.8, 3.0, 3.2, or 3.4 to about 3.5. Any range comprised of the above values ​​is expressly contemplated. The acidifying agent is typically hydrochloric acid. Digestive (gastric) enzymes, such as proteases (e.g., pepsinogen I, II, and III, pepsin, and rennin) and lipase (gastric lipase), are also present. The term "gastric juice" can refer to the liquid contents of the stomach lumen, which can be considered to have the same physical and chemical properties.

[0051] In certain embodiments, the reference gastric environment is a reference gastric environment of a subject undergoing treatment.

[0052] Thus, "potentially harmful gastric environment effects" refers to any negative impact these conditions may have on the physical, chemical, or pharmaceutical properties of the polypeptide therapeutic agent, alginate oligomer, and / or gastrointestinal permeation enhancer or on the structural integrity of the dosage form in the absence of any protection or mitigation. In the case of a polypeptide therapeutic agent, alginate oligomer, and / or gastrointestinal permeation enhancer, this includes denaturation, deterioration, precipitation, gelation, or inactivation / reduction of pharmaceutical activity of the treated agent for any reason. In the case of a dosage form, this includes dissolution, dispersion, disintegration, breakup, fragmentation, degradation, or any reduction or disruption in structural cohesion or integrity. Protection from such effects may occur by any mechanism, including physical shielding, inhibition of gastric enzyme activity, and / or local neutralization of acidic conditions.

[0053] In certain embodiments, the characteristic of "substantial protection from the stomach environment" may be considered to be met if a small percentage of the target agent denatures, degrades, precipitates, caking, or becomes inactivated / reduced in activity within about 240 minutes of entering the stomach, e.g., within about 220, 200, 180, 160, 140, 120, 100, 80, 60, 40, or 30 minutes of entering the stomach.

[0054] Similarly, in certain embodiments, the characteristic of "substantial protection from the gastric environment" may be considered to be met if a small percentage of the dosage form is lost due to dissolution, dispersion, disintegration, breakup, fragmentation, decomposition, degradation, or any type of reduction or disruption of structural cohesion or integrity within about 240 minutes of entry into the stomach, e.g., within about 220, 200, 180, 160, 140, 120, 100, 80, 60, 40, or 30 minutes of entry into the stomach.

[0055] In certain embodiments, the denaturation, degradation, precipitation, gelation, or inactivation / reduction of pharmaceutical or physiological activity of the treated agent is irreversible. In contrast, reversible denaturation, degradation, precipitation, gelation, or inactivation / reduction of pharmaceutical or physiological activity of the treated agent means that the treated agent can return to its essentially normal structure and exhibit essentially normal activity (physiological / pharmaceutical) when transferred to a less chemically extreme environment, such as the blood circulation or other neutral or mild body fluids. Expressed numerically, a return to essentially normal activity may be considered to be a recovery of at least 80%, e.g., at least 85%, 90%, 95%, or 95% of the relevant physiological or pharmacological activity of the same amount of untreated agent.

[0056] In certain embodiments, the characteristic of "substantial protection from the gastric environment" may be considered met if a small percentage of the target drug denatures, degrades, precipitates, caking, or becomes inactivated / degraded within about 240 minutes of exposure to USP simulated gastric fluid (consisting of 2.0 g NaCl, 3.2 g purified pepsin (having an activity of 800 to 2500 units / mg protein), and HCl (to a pH of 1.2) in 1000 ml water at 37°C with agitation at 100 rpm, e.g., within about 220, 200, 180, 160, 140, 120, 100, 80, 60, 40, or 30 minutes of exposure to simulated gastric fluid at such conditions. Dissolution testing may be performed in United States Pharmacopeia (USP) Apparatus I (100 rpm).

[0057] In certain embodiments, the characteristic of "substantial protection from the gastric environment" may be considered met if a small percentage of the dosage form is lost due to dissolution, dispersion, disintegration, breakup, fragmentation, degradation, or any type of reduction or disruption of structural cohesion or integrity within about 240 minutes of exposure to USP simulated gastric fluid (consisting of 2.0 g NaCl, 3.2 g purified pepsin (having an activity of 800 to 2500 units / mg protein), and HCl (to a pH of 1.2) in 1000 ml of water at 37°C with agitation at 100 rpm, e.g., within about 220, 200, 180, 160, 140, 120, 100, 80, 60, 40, or 30 minutes of exposure to simulated gastric fluid at such conditions. Dissolution testing may be performed in United States Pharmacopeia (USP) Apparatus I (100 rpm).

[0058] In certain embodiments, the methods described above comprise administering the polypeptide therapeutic agent together with an alginate oligomer, and optionally a gastrointestinal permeation enhancer, to the stomach of a human subject or non-human animal subject in one or more dosage forms, wherein the polypeptide therapeutic agent, alginate oligomer, and, if used, the gastrointestinal permeation enhancer are not administered as part of one or more dosage forms structured to provide substantial protection from the gastric environment to the polypeptide therapeutic agent, alginate oligomer, and / or gastrointestinal permeation enhancer that form part of the dosage form.

[0059] Alternatively, the polypeptide therapeutic agent, alginate oligomer and / or gastrointestinal permeation enhancer are administered to the stomach in a manner (e.g., as part of a susceptible dosage form) that renders them significantly susceptible to the effects of the potentially harmful gastric environment, e.g., in a manner that allows or enables exposure to the gastric environment and / or the effects of the potentially harmful gastric environment.

[0060] This may be described as a substantial proportion of the drug, or dosage form of which the drug is a part, being substantially susceptible to instability in the gastric environment (e.g., denaturation, deterioration, precipitation, caking, or inactivation / reduction in activity in some way in the case of a drug, or dissolution, dispersion, or disintegration in the case of a dosage form) while it remains in the stomach to which it is administered, which may further be described as unprotected.

[0061] In certain embodiments, the dosage form is one that substantially loses all bonds or structural integrity, e.g., substantially dissolves, disperses, disintegrates, breaks up, fragments, degrades, or deteriorates, within about 240 minutes of entering the stomach, e.g., within about 220, 200, 180, 160, 140, 120, 100, 80, 60, 40, 30, 20, 15, 10, or 5 minutes of entering the stomach.

[0062] Similarly, in certain embodiments, the dosage form is one that allows a substantial proportion of the target agent to denature, degrade, precipitate, caking, or become inactivated / degraded within about 240 minutes of entering the stomach, e.g., within about 220, 200, 180, 160, 140, 120, 100, 80, 60, 40, 30, 20, 15, 10, or 5 minutes of entering the stomach.

[0063] In certain embodiments, the dosage form is one that substantially loses all cohesiveness or structural integrity (i.e., a substantial proportion of the dosage form loses cohesiveness or structural integrity), e.g., substantially dissolves, disperses, disintegrates, breaks up, fragments, degrades, or deteriorates, within about 240 minutes of exposure to USP simulated gastric fluid (consisting of 2.0 g NaCl, 3.2 g purified pepsin (having an activity of 800-2500 units / mg protein), and HCl (to a pH of 1.2) in 1000 ml of water at 37°C with agitation at 100 rpm, e.g., within about 220, 200, 180, 160, 140, 120, 100, 80, 60, 40, 30, 20, 15, 10, or 5 minutes of exposure to simulated gastric fluid at such conditions. Dissolution testing may be performed in United States Pharmacopeia (USP) Apparatus I (100 rpm).

[0064] Similarly, in certain embodiments, the dosage form is one that allows a substantial proportion of the target drug to denature, degrade, precipitate, caking, or become inactivated / inactive within about 240 minutes of exposure to USP simulated gastric fluid (consisting of 2.0 g NaCl, 3.2 g purified pepsin (having an activity of 800-2500 units / mg protein), and HCl (to a pH of 1.2) in 1000 ml of water at 37°C with agitation at 100 rpm, e.g., within about 220, 200, 180, 160, 140, 120, 100, 80, 60, 40, 30, 20, 15, 10, or 5 minutes of exposure to simulated gastric fluid at such conditions. Dissolution testing may be performed in United States Pharmacopeia (USP) Apparatus I (100 rpm).

[0065] The dosage form for use in the present invention can be any configuration common to oral, oral gavage, nasogastric gavage, or intragastric administration techniques. Thus, the dosage form can be any pharmaceutically acceptable composition, for example, in the form of a solution, dispersion, emulsion, powder, tablet, capsule, gel, etc. Conveniently, the dosage form is in the form of a tablet or a capsule filled with liquid or powder. These are considered examples of solid dosage forms. In certain embodiments, the dosage form has dimensions that allow it to be swallowed by the subject receiving treatment.

[0066] In certain embodiments, the dosage form does not comprise, e.g., is substantially free of, alginate oligomers in solid form, e.g., gelling or partially gelling form. Specifically, in certain embodiments, the therapeutic polypeptide is not encapsulated by or otherwise associated with alginate oligomers in solid form, e.g., gelling or partially gelling form, such that the therapeutic polypeptide is substantially protected from the gastric environment. In certain embodiments, the dosage form is provided such that, after entry into the stomach, no alginate oligomers in the dosage form encapsulate or otherwise associate with the polypeptide therapeutic agent, e.g., in solid form, particularly in gelling or partially gelling form, such that the therapeutic polypeptide is substantially protected from the gastric environment.

[0067] In certain embodiments, the various types of components of the dosage form are not cross-linked to themselves and / or to each other. Such cross-links may be covalent or ionic, e.g., involving divalent cations. In certain embodiments, the alginate oligomer molecules in the dosage form are not cross-linked to each other. In certain embodiments, the alginate oligomer molecules in the dosage form are not cross-linked to other components in the dosage form, e.g., polypeptide therapeutic agents.

[0068] In certain embodiments, the polypeptide therapeutic agent and the alginate oligomer are not provided in the dosage form as a pre-formed stable non-covalent complex.

[0069] In certain embodiments, the polypeptide therapeutic agent is not present in its granular form as part of the dosage form. In certain embodiments, the alginate oligomer is not present in its granular form in the dosage form. In other embodiments, the polypeptide therapeutic agent and / or the alginate oligomer and / or the gastrointestinal permeation enhancer are not present in or on particles as part of the dosage form. In other embodiments, the dosage form does not comprise particles.

[0070] The above references to particles include microparticles and nanoparticles. A microparticle may be considered to be any particle having a particle size in the micrometer range, i.e., from about 1 μm to about 1000 μm, for example, from about 1 μm to about 900 μm, 800 μm, 700 μm, 600 μm, 500 μm, 400 μm, 300 μm, 200 μm, 100 μm, 50 μm, 40 μm, 30 μm, 20 μm, 10 μm, or 5 μm, or from about 1 μm, 5 μm, 10 μm, 20 μm, 30 μm, 40 μm, 50 μm, 100 μm, 200 μm, 300 μm, 400 μm, 500 μm, 600 μm, 700 μm, 800 μm, or 900 μm to about 1000 μm. A nanoparticle may be considered any particle having a particle size in the nanometer range, i.e., from about 1 nm to about 1000 nm, e.g., from about 1 nm to about 900 nm, 800 nm, 700 nm, 600 nm, 500 nm, 400 nm, 300 nm, 200 nm, 100 nm, or 50 nm, or from about 50 nm, 100 nm, 200 nm, 300 nm, 400 nm, 500 nm, 600 nm, 700 nm, 800 nm, or 900 nm to about 1000 nm. Any ranges having endpoints formed from any of the above values ​​are also expressly disclosed. In the context of nanoparticles, the term "particle size" refers to the size of the particle as measured using dynamic light scattering (e.g., quasi-elastic light scattering). For example, particle size can be measured using dynamic light scattering instruments (e.g., the Zetasizer Nano ZS model manufactured by Malvern Instruments Ltd. and the ELS-8000 manufactured by Otsuka Electronics Co., Ltd.). These instruments measure the Brownian motion of particles, and particle size is measured based on established dynamic light scattering theory. In the context of microparticles, the term "particle size" refers to the size of the particle as measured using laser diffraction spectroscopy. Commercially available instruments include the Mastersizer 3000 instrument manufactured by Malvern Instruments.

[0071] In certain embodiments, the dosage form is not bioadhesive and does not include a bioadhesive component. In certain embodiments, the dosage form is not mucoadhesive and does not include a mucoadhesive component.

[0072] The polypeptide therapeutic agent, alginate oligomer, and / or gastrointestinal permeation enhancer may be located within and / or on any portion of the dosage form, either together or separately. They may be intimately intermixed or present in multiple discrete compartments, e.g., layers. Indeed, it is expressly contemplated that multilayer dosage forms may be provided in which an alginate oligomer-containing layer surrounds an inner compartment or layer containing the polypeptide therapeutic agent and / or gastrointestinal permeation enhancer. A further configuration is a multilayer dosage form having an alginate oligomer-containing outer layer, an innermost compartment or layer containing the polypeptide therapeutic agent, and an intermediate layer containing the gastrointestinal permeation enhancer. All other physical configurations of these components, including alternating layers / compartments of two or more components, are expressly contemplated. The presence of excipients, such as any of those listed herein, along with the polypeptide therapeutic agent, alginate oligomer, and / or gastrointestinal permeation enhancer, if present, is also expressly contemplated. In these embodiments, the excipients may vary between layers / compartments. In yet another arrangement, there may be a layer of excipient, such as an outer coating layer.

[0073] The dosage forms of which the therapeutic polypeptide, alginate oligomer and optionally gastrointestinal permeation enhancer for use in the present invention are a part do not include a coating (or barrier, layer or film) that provides substantial protection from the gastric environment in addition to the polypeptide therapeutic, alginate oligomer and, if used, the gastrointestinal permeation enhancer.

[0074] Such a coating would be a layer of material, typically an outer or outermost layer, in or on the dosage form that provides substantial protection from the gastric environment to the portions of the dosage form internal to the coating, e.g., the polypeptide therapeutic agent, alginate oligomer, and / or, if used, the gastrointestinal permeation enhancer. Such a coating would be formed from a material that is protective against the gastric environment in addition to the polypeptide therapeutic agent, alginate oligomer, and, if used, the gastrointestinal permeation enhancer, although the presence of the polypeptide therapeutic agent, alginate oligomer, and, if used, the gastrointestinal permeation enhancer in the coating is not precluded. In other embodiments, the coating is not or does not include an alginate oligomer.

[0075] Thus, these compositions do not have a so-called "enteric coating," i.e., a coating (or barrier, layer, or film) that remains substantially undegraded in a subject's stomach for the time it takes for such compositions to pass through the stomach. Enteric coatings are insoluble in the stomach and prevent the passage of gastric juice components, such as acid.

[0076] Such coatings are typically prepared from polymers containing fatty acids, waxes, shellac, plastics, and plant fibers. Specific examples include, but are not limited to, methyl acrylate-methacrylic acid copolymer, methyl methacrylate-methacrylic acid copolymer, cellulose acetate succinate, cellulose acetate phthalate, hydroxypropyl methylcellulose phthalate, hydroxypropyl methylcellulose acetate succinate (hypromellose acetate succinate), polyvinyl acetate phthalate (PVAP), cellulose acetate trimellitate, and those available under the trade name Eudragit, and alginate polymers (i.e., alginates greater than 30, 25, 20, or 15 kDa). Such coatings may be applied in conventional manners and at conventional thicknesses / amounts.

[0077] In certain embodiments, the dosage form and / or its components, eg, alginate oligomers, are not conjugated with polyethylene glycol polymers (PEGylated).

[0078] In certain embodiments, the dosage form may comprise a therapeutic polypeptide, an alginate oligomer, and / or a gastrointestinal permeation enhancer.

[0079] It will be appreciated that the features of the invention as defined above should not be construed as excluding the possibility that the structure of the dosage form will still provide protection for the targeted agent or dosage form of which it is a part from the gastric environment and / or the deleterious effects of the gastric environment for a limited period of time or to a limited extent, for example for up to 60 minutes or for up to 55, 50, 45, 40, 35, 30, 25, 20, 10, or 5 minutes from entry into the stomach, but that subsequent exposure to the gastric environment and / or the deleterious effects of the gastric environment may occur prior to completion of gastric transit / retention / retention of the targeted agent or dosage form of which it is a part.

[0080] In certain specific embodiments, the dosage form may have a structure in which the alginate oligomer surrounds the polypeptide therapeutic agent, providing the polypeptide therapeutic agent with the limited protection described above for the time required for the dosage form to reach the surface of the gastric mucosa.

[0081] "Used together" or "administered together" or similar expressions means that at least two therapeutically active agents (i.e., a therapeutic polypeptide and an alginate oligomer), or at least three (if a gastrointestinal permeation enhancer is also used), are used in combination to achieve the effect of interest (e.g., increased systemic bioavailability, gastric uptake, absorption from the stomach, improved efficacy of a therapeutic polypeptide in treating or preventing a disease or condition responsive to a therapeutic polypeptide, or a complication thereof).

[0082] It particularly refers to administration of an effective (e.g., pharmaceutically effective) amount of alginate oligomer simultaneously or substantially simultaneously with, or before, an effective (e.g., pharmaceutically effective) amount of a therapeutic polypeptide, and, optionally, an effective (e.g., pharmaceutically effective) amount of a gastrointestinal permeation enhancer. In other embodiments, the oligomer is administered separately, before and after, the therapeutic polypeptide and / or the gastrointestinal permeation enhancer. One skilled in the art will readily be able to design a dosing regimen to maximize the effectiveness of the alginate oligomer and therapeutic polypeptide, and, optionally, the gastrointestinal permeation enhancer, used in the methods of the present invention. "Used together" does not implicitly indicate that the respective agents are present in the same dosage form, formulation, or composition; therefore, even when used or administered simultaneously or substantially simultaneously, the alginate oligomer and therapeutic polypeptide, and, optionally, the gastrointestinal permeation enhancer, need not be present in the same dosage form, composition, or formulation, but may be administered separately. Thus, "separate" use / administration includes use / administration at the same or substantially the same time or at different times, e.g., sequentially or with various time intervals, according to the desired dosage or regime. Thus, "simultaneous" administration includes administration of the alginate oligomer and therapeutic polypeptide, and optionally the gastrointestinal permeation enhancer, within the same dosage form composition or formulation, or within separate compositions / formulations administered simultaneously or substantially simultaneously. When at least three agents are used according to the present invention, two (e.g., the alginate oligomer and therapeutic polypeptide, or the alginate oligomer and gastrointestinal permeation enhancer, or the therapeutic polypeptide and the gastrointestinal permeation enhancer) may be administered in one dosage form / composition and the third in another. It may be advantageous to administer all three agents in a single dosage form composition, e.g., as a solution or in tablet or capsule form.

[0083] The term "substantially simultaneously" includes administration of the therapeutic polypeptide immediately before, immediately after, or nearly immediately before or after the alginate oligomer and / or, if desired, immediately before, immediately after, or nearly immediately before or after the gastrointestinal permeation enhancer. The term "almost immediately" may be read to include administration within 1 hour, preferably within 30, 20, 10, 5, 4, 3, 2, or 1 minute, of the previous administration. The therapeutic polypeptide may be administered in multiple applications before, with, or after the alginate oligomer and, if desired, the gastrointestinal permeation enhancer. The alginate oligomer may be administered in multiple applications before, with, or after the therapeutic polypeptide and, if desired, the gastrointestinal permeation enhancer.

[0084] Thus, the present invention further provides a dosage form adapted for delivery to the stomach of a human subject, or a non-human animal subject, said composition comprising: (i) a polypeptide therapeutic; and (ii) an alginate oligomer; (iii) a gastrointestinal permeation enhancer; and wherein the dosage form does not have a coating that provides substantial protection from the gastric environment in addition to the polypeptide therapeutic agent, the alginate oligomer, and the gastrointestinal permeation enhancer.

[0085] The above detailed discussion of dosage forms of the present invention applies mutatis mutandis to this aspect of the invention.

[0086] The dosage form comprising the therapeutic polypeptide, alginate oligomer, and optionally a gastrointestinal permeation enhancer used in the present invention may be administered to the stomach of the subject undergoing treatment by oral, oral gastric tube, nasogastric tube, or intragastric route. The same or different routes may be used for the various dosage forms administered by the present invention. Using the same route may be convenient. Oral administration is preferred due to its ease and comfort for the subject, thus increasing the likelihood of compliance.

[0087] It will be readily apparent that the methods of the present invention do not further involve the separate administration of the polypeptide therapeutic agent, the alginate oligomer and, if used, the gastrointestinal permeation enhancer, or more generally the use of a compound or device capable of protecting the dosage form from the gastric environment.

[0088] Gastric uptake of an active or reversibly reduced polypeptide therapeutic agent and / or absorption of an active or reversibly reduced polypeptide therapeutic agent from the stomach may be measured by analyzing the plasma level of the active or reversibly reduced polypeptide therapeutic agent at a time before gastric emptying. Alternatively or additionally, these phenomena may be measured by analyzing the comparative plasma levels of the active or reversibly reduced polypeptide therapeutic agent in the splenic vein, which drains the gastric cavity, and the portal vein, which drains the gastrointestinal system. Gastric uptake / absorption following administration to the stomach will indicate an increase in plasma concentration in the splenic vein compared to the portal vein.

[0089] The methods of the invention may include measuring the gastric uptake of the polypeptide therapeutic agent, or an active or reversibly reduced form of the polypeptide therapeutic agent, and / or measuring the absorption of the polypeptide therapeutic agent, or an active or reversibly reduced form of the polypeptide therapeutic agent, from the stomach after administration of the polypeptide therapeutic agent together with the alginate oligomer and, optionally, a gastrointestinal permeation enhancer. Any of the measurement techniques described above may be performed. These measurements may be performed within 360, 330, 300, 270, 240, 210, 180, 120, 60, 30, 20, 15, 10, or 5 minutes of administration of the last administered polypeptide therapeutic agent, alginate oligomer, or gastrointestinal permeation enhancer.

[0090] According to the present invention, gastric uptake / absorption from the stomach refers to a polypeptide therapeutic agent in an active form or in a reduced-activity form that may be reversed. This means that the polypeptide therapeutic agent is taken up from the gastric environment into the blood, e.g., via the splenic vein, in a form that retains the inherent therapeutic activity of the polypeptide or that reverts to a form that exhibits the inherent therapeutic activity of the polypeptide upon entering the subject's bloodstream or by the time it reaches its site of therapeutic action. In other words, the polypeptide therapeutic agent is taken up from the gastric environment into the blood, e.g., via the splenic vein, in a form that may be therapeutically effective or that reverts to a therapeutically effective form upon entering the subject's bloodstream or by the time it reaches its site of therapeutic action. Thus, the polypeptide therapeutic agent is taken up in a substantially undegraded, unreduced / inactive, and / or undenatured form, or at least in a form that is not irreversibly reduced / inactive and / or undenatured.

[0091] As noted above, alginates typically exist as polymers of at least 35,000 daltons average molecular mass, i.e., approximately 175 to approximately 190 monomer residues, although typically much higher, and alginate oligomers according to the present invention may be defined as materials obtained by fractionation (i.e., size reduction) of alginate polymers, usually native alginate. Alginate oligomers may be considered to be alginates of average molecular weight less than 15,000 daltons (i.e., approximately less than 100 monomer residues).

[0092] Viewed another way, oligomers for use according to the invention will typically contain from 2 to 100, more typically from 3, 4, 5 or 6 to 100 monomer residues, and may contain from 2, 3, 4, 5, or 6 to 75, 2, 3, 4, 5, or 6 to 50, 2, 3, 4, 5, or 6 to 40, 2, 3, 4, 5, or 6 to 35, or 2, 3, 4, 5, or 6 to 30 residues. Thus, alginate oligomers for use according to the invention will typically have an average molecular weight of from 350, 550, 700, 900, or 1000 to 15,000 daltons, from 350, 550, 700, 900, or 1000 to 10,000 daltons, from 350, 550, 700, 900, or 1000 to 8000 daltons, from 350, 550, 700, 900, or 1000 to 7000 daltons, or from 350, 550, 700, 900, or 1000 to 6,000 daltons.

[0093] In other words, the alginate oligomer may have a degree of polymerization (DP) or number average degree of polymerization (DPn) of from 2 to 100, preferably from 2 to 75, preferably from 2 to 50, more preferably from 2 to 40, 2 to 35, 2 to 30, 2 to 28, 2 to 25, 2 to 22, 2 to 20, 2 to 18, 2 to 17, 2 to 15, or 2 to 12.

[0094] Other representative ranges (whether number of residues, DP, or DPn) include any one of 3, 4, 5, 6, 7, 8, 9, 10, or 11 to any one of 50, 45, 40, 39, 38, 37, 36, 35, 34, 33, 32, 31, 30, 29, 28, 27, 26, 25, 24, 23, 22, 21, 20, 19, 18, 17, 16, 15, 14, 13, or 12.

[0095] Other representative ranges (whether number of residues, DP, or DPn) include any one of 8, 9, 10, 11, 12, 13, 14, or 15 to any one of 50, 45, 40, 39, 38, 37, 36, 35, 34, 33, 32, 31, 30, 29, 28, 27, 26, 25, 24, 23, 22, 21, 20, 19, 18, 17, or 16.

[0096] Other representative ranges (whether number of residues, DP, or DPn) include any one of 11, 12, 13, 14, 15, 16, 17, or 18 to any one of 50, 45, 40, 39, 38, 37, 36, 35, 34, 33, 32, 31, 30, 29, 28, 27, 26, 25, 24, 23, 22, 21, 20, or 19.

[0097] Generally speaking, alginate oligomers, as defined above, contain (or comprise) guluronate or guluronic acid (G) and / or mannuronate or mannuronic acid (M) residues or units. Alginate oligomers according to the present invention preferably consist exclusively or substantially exclusively (i.e., consist essentially of) uronate / uronic acid residues, more particularly G and M residues or G residues, so long as at least 30% of the monomer residues are G residues. In other words, in alginate oligomers for use in the present invention, at least 80%, more particularly at least 85, 90, 95, or 99% of the monomer residues may be uronate / uronic acid residues, or more particularly G and M residues or G residues, so long as at least 30% of the monomer residues are G residues. In other words, preferably, the alginate oligomer does not contain other residues or units (e.g., other sugar residues, or, more particularly, other uronic acid / uronate residues).

[0098] The alginate oligomer is preferably a linear oligomer.

[0099] As previously indicated, at least 30% of the monomer residues of an alginate oligomer are G residues (i.e., guluronate or guluronic acid). In other words, the alginate oligomer comprises at least 30% guluronate (or guluronic acid) residues. Accordingly, particular embodiments include alginate oligomers having (e.g., containing) 30 to 70% G (guluronate) residues or 70 to 100% G (guluronate) residues. Thus, a representative alginate oligomer for use according to the present invention may comprise at least 70% G residues (i.e., at least 70% of the monomer residues of the alginate oligomer are G residues).

[0100] Preferably, at least 40%, 45%, 50%, 55%, or 60%, more particularly at least 70% or 75%, and even more particularly at least 80, 85, 90, 91, 92, 93, 94, 95, 96, 97, 98, or 99% of the monomer residues are guluronate. In one embodiment, the alginate oligomer may be an oligoguluronate (i.e., a homo-oligomer of G, or 100% G).

[0101] In a further preferred embodiment, the alginates of the invention described above have a primary structure in which the majority of G residues are in so-called G-blocks. Preferably, at least 50%, more preferably at least 70 or 75%, and most preferably at least 80, 85, 90, 92, or 95% of the G residues are in G-blocks. A G-block is a string of at least two adjacent G residues, preferably at least three adjacent G residues, more preferably at least four or five adjacent G residues, and most preferably at least seven adjacent G residues.

[0102] Specifically, at least 90% of the G residues are in 1-4 linkages with other G residues, more specifically at least 95%, more preferably at least 98%, and most preferably at least 99% of the G residues in the alginate are in 1-4 linkages with other G residues.

[0103] The alginate oligomers used in the present invention are preferably 3-35mers, more preferably 3-28mers, particularly 4-25mers, such as 5-20mers, particularly 6-22mers, particularly 8-20mers, and particularly 10-15mers, and have molecular weights ranging from, for example, 350 to 6400 daltons, or 350 to 6000 daltons, preferably 550 to 5500 daltons, preferably 750 to 5000 daltons, and particularly 750 to 4500 daltons, or 2000 to 3000 daltons, or 900 to 3500 daltons. Other representative alginate oligomers include those having 5, 6, 7, 8, 9, 10, 11, 12, or 13 to 50, 45, 40, 35, 28, 25, 22, or 20 residues, as described above.

[0104] The alginic acid oligomer of the present invention has a degree of polymerization (DP) or number average degree of polymerization (DP) of 3 to 28, 4 to 25, 6 to 22, 8 to 20, or 10 to 15, or 5 to 18, or 7 to 15, or 8 to 12, particularly 10. n ) may be present.

[0105] The alginate trigomers of the present invention may have a degree of polymerization (DP) of 3 to 24, 4 to 23, 5 to 22, 6 to 21, 7 to 20, 8 to 19, 9 to 18, 10 to 17, 11 to 16, 12 to 15, or 13 to 14 (e.g., 13 or 14), or a number average degree of polymerization (DP n ) may be present.

[0106] The alginic acid oligomer of the present invention may have a degree of polymerization (DP) or number average degree of polymerization (DP) of 4 to 25, 5 to 24, 6 to 23, 7 to 22, 8 to 21, 9 to 20, 10 to 19, 11 to 18, 12 to 17, 13 to 16, 14 to 15 (e.g., 14 or 15). n ) may be present.

[0107] The alginate oligomer of the present invention may have a degree of polymerization (DP) or number average degree of polymerization (DP) of 5 to 26, 6 to 25, 7 to 24, 8 to 23, 9 to 22, 10 to 21, 11 to 20, 12 to 19, 13 to 18, 14 to 17, or 15 to 16 (e.g., 15 or 16). n) may be present.

[0108] The alginic acid oligomer of the present invention has a degree of polymerization (DP) or number average degree of polymerization (DP) of 4 to 50, 4 to 40, 4 to 35, 4 to 30, 4 to 28, 4 to 26, 4 to 22, 4 to 20, 4 to 18, 4 to 16, or 4 to 14. n ) may be present.

[0109] The alginic acid oligomer of the present invention has a degree of polymerization (DP) or number average degree of polymerization (DP) of 5 to 50, 5 to 40, 5 to 25, 5 to 22, 5 to 20, 5 to 18, 5 to 23, 5 to 20, 5 to 18, 5 to 16, or 5 to 14. n ) may be present.

[0110] The alginic acid oligomer of the present invention has a degree of polymerization (DP) or number average degree of polymerization (DP) of 6 to 50, 6 to 40, 6 to 35, 6 to 30, 6 to 28, 6 to 26, 6 to 24, 6 to 20, 6 to 19, 6 to 18, 6 to 16, or 6 to 14. n ) may be present.

[0111] The alginic acid oligomer of the present invention has a degree of polymerization (DP) or number average degree of polymerization (DP) of 8 to 50, 8 to 40, 8 to 35, 8 to 30, 8 to 28, 8 to 25, 8 to 22, 8 to 20, 8 to 18, 8 to 16, or 8 to 14. n ) may be present.

[0112] The alginic acid oligomer of the present invention has a degree of polymerization (DP) or number average degree of polymerization (DP) of 9 to 50, 9 to 40, 9 to 35, 9 to 30, 9 to 28, 9 to 25, 9 to 22, 9 to 20, 9 to 18, 9 to 16, or 9 to 14. n ) may be present.

[0113] The alginate oligomer of the present invention has a degree of polymerization (DP) or number average degree of polymerization (DP) of 10 to 50, 10 to 40, 10 to 35, 10 to 30, 10 to 28, 10 to 25, 10 to 22, 10 to 20, 10 to 18, 10 to 16, or 10 to 14. n ) may be present.

[0114] The alginic acid oligomer of the present invention has a degree of polymerization (DP) or number average degree of polymerization (DP) of 11 to 50, 11 to 40, 11 to 35, 11 to 30, 11 to 28, 11 to 25, 11 to 22, 11 to 20, 11 to 18, 11 to 16, or 11 to 14. n ) may be present.

[0115] The alginate oligomer of the present invention has a degree of polymerization (DP) or number average degree of polymerization (DP) of 12 to 50, 12 to 40, 12 to 35, 12 to 30, 12 to 28, 12 to 25, 12 to 22, 12 to 20, 12 to 18, 12 to 16, or 12 to 14. n ) may be present.

[0116] The alginic acid oligomer of the present invention has a degree of polymerization (DP) or number average degree of polymerization (DP) of 13 to 50, 13 to 40, 13 to 35, 13 to 30, 13 to 28, 13 to 25, 13 to 22, 13 to 20, 13 to 18, 13 to 16, or 13 to 14. n ) may be present.

[0117] The alginic acid oligomer of the present invention has a degree of polymerization (DP) or number average degree of polymerization (DP) of 14 to 50, 14 to 40, 14 to 35, 14 to 30, 14 to 28, 14 to 25, 14 to 22, 14 to 20, 14 to 18, 14 to 16, or 14 to 15. n ) may be present.

[0118] The alginate oligomer of the present invention has a degree of polymerization (DP) or number average degree of polymerization (DP) of 15 to 50, 15 to 40, 15 to 35, 15 to 30, 15 to 28, 15 to 25, 15 to 22, 15 to 20, 15 to 18, or 15 to 16. n ) may be present.

[0119] The alginate oligomer of the present invention has a degree of polymerization (DP) or number average degree of polymerization (DP) of 18 to 50, 18 to 40, 18 to 35, 18 to 30, 18 to 28, 18 to 25, 18 to 22, or 18 to 20. n ) may be present.

[0120] Preferably, the alginate oligomers of the present invention are substantially free, preferably essentially free, of alginate oligomers having a degree of polymerization outside the ranges disclosed herein. This may be expressed as the molecular weight distribution of the alginate oligomers of the present invention, e.g., the percentage of each mole of alginate oligomer used in accordance with the present invention that has a DP outside the conforming range. The molecular weight distribution preferably has no more than 10%, preferably no more than 9, 8, 7, 6, 5, 4, 3, 2, or 1% of the moles having a conforming DP. n Similarly, 10% or less, preferably 9, 8, 7, 6, 5, 4, 3, 2, or 1% molar or less of the DP is acceptable. n It is preferred to have a DP that is 3, 2, or 1 less than the lower limit for

[0121] Suitable alginate oligomers are described in WO 2007 / 039754, WO 2007 / 039760, WO 2008 / 125828 and WO 2009 / 068841, the disclosures of which are expressly incorporated herein by reference in their entireties.

[0122] Representative suitable alginate oligomers have a DP in the range of 5 to 30. n , a guluronate / galacturonate ratio (F) of at least 0.80 G ), mannuronate ratio (F) of 0.20 or less M ), and at least 95 mole % of the DP is 25 or less.

[0123] Further suitable alginic acid oligomers have a number average degree of polymerization ranging from 7 to 15 (preferably from 8 to 12), a guluronate / galacturonate ratio (F) of at least 0.85 (preferably at least 0.90). G ), a mannuronate ratio (F) of 0.15 or less (preferably 0.10 or less). M ), with at least 95% of the moles having a degree of polymerization of less than 17 (preferably less than 14).

[0124] Further suitable alginic acid oligomers have a number average degree of polymerization ranging from 5 to 18 (in particular from 7 to 15), a guluronate / galacturonate ratio (F) of at least 0.80 (preferably at least 0.85, in particular at least 0.92). G ), a mannuronate ratio (F) of 0.20 or less (preferably 0.15 or less, particularly 0.08 or less). M ), with at least 95% of the moles having a degree of polymerization of less than 20 (preferably less than 17).

[0125] Further suitable alginate oligomers have a number average degree of polymerization ranging from 5 to 18, a guluronate / galacturonate ratio (F) of at least 0.92. G ), mannuronate ratio of 0.08 or less (F M ) and at least 95% of the moles have a degree of polymerization less than 20.

[0126] Further suitable alginic acid oligomers have a number average degree of polymerization in the range of from 5 to 18 (preferably from 7 to 15, more preferably from 8 to 12, in particular about 10), a guluronate / galacturonate ratio (F) of at least 0.80 (preferably at least 0.85, more preferably at least 0.90, in particular at least 0.92, especially at least 0.95). G ), a mannuronate ratio (F) of 0.20 or less (preferably 0.15 or less, more preferably 0.10 or less, particularly 0.08 or less, and especially 0.05 or less). M ), with at least 95% of the moles having a degree of polymerization of less than 20 (preferably less than 17, more preferably less than 14).

[0127] Further suitable alginic acid oligomers have a number average degree of polymerization ranging from 7 to 15 (preferably from 8 to 12), a guluronate / galacturonate ratio (F) of at least 0.92 (preferably at least 0.95). G ), a mannuronate fraction (F) of 0.08 or less (preferably 0.05 or less). M ), with at least 95% of the moles having a degree of polymerization of less than 17 (preferably less than 14).

[0128] Further suitable alginic acid oligomers have a number average degree of polymerization ranging from 5 to 18, a guluronate / galacturonate ratio (F) of at least 0.80. G ), mannuronate ratio (F) of 0.20 or less M ) and at least 95% of the moles have a degree of polymerization less than 20.

[0129] Further suitable alginate oligomers have a number average degree of polymerization ranging from 7 to 15, a guluronate / galacturonate ratio (F) of at least 0.85. G ), mannuronate fraction (F) of 0.15 or less M ), and at least 95% of the moles have a degree of polymerization less than 17.

[0130] Further suitable alginic acid oligomers have a number average degree of polymerization ranging from 7 to 15, a guluronate / galacturonate ratio (F) of at least 0.92. G ), mannuronate fraction (F) of 0.08 or less M ), and at least 95% of the moles have a degree of polymerization less than 17.

[0131] Further suitable alginate oligomers have a number average degree of polymerization ranging from 5 to 20, a guluronate ratio (F) of at least 0.85. G ) and a mannuronate ratio (F) of 0.15 or less M )

[0132] Thus, in accordance with the present invention, it is recognized that a particular type of advantageous alginate oligomer is the so-called "high G" or "G-block" oligomer, i.e., alginate oligomer defined as having a high content of G residues or G-blocks (e.g., at least 70% of the monomer residues are G, preferably arranged in G-blocks). However, as further described below, other types of alginate oligomers may also be used, including in particular "high M" or "M-block" oligomers or M-block oligomers. Thus, it is an alginate oligomer in which a high proportion of a single monomer type is present, with the majority of said monomers of this type present in adjacent rows of that monomer type, representing particularly preferred oligomers, e.g., oligomers in which at least 70% of the monomer residues in the oligomer are G residues in 1-4 linkage with another G residue, or more preferably, at least 75%, and most preferably, at least 80, 85, 90, 92, 93, 94, 95, 96, 97, 98, 99% of the monomer residues in the oligomer are G residues in 1-4 linkage with another G residue. Alternatively, this 1-4 linkage of two G residues can be expressed as a guluronic unit linked to adjacent guluronic units.

[0133] In further embodiments, at least 50%, or more particularly, more than 50% of the monomer residues of the alginate oligomer may be M residues (i.e., mannuronate or mannuronic acid). In other words, the alginate oligomer comprises at least, or more than 50%, mannuronate (or mannuronic acid) residues. Specific embodiments therefore include alginate oligomers having (e.g., containing) 50 to 70% M (mannuronate) residues, or, for example, 70 to 100% M (mannuronate) residues. Further specific embodiments also include oligomers containing 71 to 85% M residues, or 85 to 100% M residues. Thus, a typical alginate oligomer for use according to this embodiment of the invention will comprise more than 70% M residues (i.e., more than 70% of the monomer residues of the alginate oligomer are M residues).

[0134] In other embodiments, at least 50%, or 60%, more particularly at least 70%, or 75%, and even more particularly at least 80, 85, 90, 95, or 99% of the monomer residues are mannuronate. In one embodiment, the alginate oligomer may be an oligomannuronate (i.e., a homo-oligomer of M or 100% M).

[0135] In a further embodiment, the alginates of the invention described above have a primary structure in which the majority of M residues are in so-called M-blocks. In this embodiment, preferably at least 50%, more preferably at least 70 or 75%, and most preferably at least 80, 85, 90, or 95% of the M residues are in M-blocks. An M-block is an adjacent sequence of at least two M residues, preferably at least three adjacent M residues, more preferably at least four or five adjacent M residues, and most preferably at least seven adjacent M residues.

[0136] Specifically, at least 90% of the M residues are in 1-4 bonds to other M residues. More specifically, at least 95%, more preferably at least 98%, and most preferably at least 99% of the M residues in the alginate are in 1-4 bonds to other M residues.

[0137] Other preferred oligomers are alginate oligomers in which at least 70% of the monomeric residues in the oligomer are M residues linked in 1-4 bond to another M residue, or more preferably at least 75%, and most preferably at least 80, 85, 90, 92, 93, 94, 95, 96, 97, 98, 99% of the monomeric residues in the oligomer are M residues linked in 1-4 bond to another M residue. Alternatively, this 1-4 bond between two M residues can be represented as a mannuronic unit linked to adjacent mannuronic units.

[0138] In another further embodiment, the alginate oligomer of the present invention comprises a string of alternating M and G residues. A string of at least three, preferably at least four, alternating M and G residues represents an MG block. Preferably, the alginate oligomer of the present invention comprises an MG block. More specifically, an MG block is a string of at least three adjacent residues consisting of G and M residues, in which each non-terminal (internal) G residue in the string is bonded to an M residue by 1-4 and 4-1 bonds, and each non-terminal (internal) M residue in the string is bonded to a G residue by 1-4 and 4-1 bonds. Preferably, the MG block is at least five or six adjacent residues, more preferably at least seven or eight adjacent residues.

[0139] In a further embodiment, the minority uronates (i.e., mannuronate or guluronate) in the alginate oligomer are found predominantly in the MG block. In this embodiment, preferably at least 50%, more preferably at least 70 or 75%, and most preferably at least 80, 85, 90, or 95% of the minority uronate monomers in the MG block alginate oligomer are present in the MG block. In another embodiment, the alginate oligomer is arranged so that at least 50%, at least 60%, at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 99%, e.g., 100%, of the G and M residues in the oligomer are located in the MG block.

[0140] In its broadest scope, the present invention extends to embodiments in which at least 1% but less than 100% of the monomer residues of the oligomer are G residues (i.e., guluronate or guluronic acid), but more particularly, and as further defined below, at least 30% of the monomer residues are G residues. Thus, in its broadest scope, an MG block-containing alginate oligomer may contain at least 1% but less than 100% guluronate (or guluronic acid) residues, but typically an MG block-containing alginate oligomer comprises at least 30% (or at least 35, 40, or 45% or 50% G), but less than 100% G. Thus, specific embodiments include MG block-containing alginate oligomers having (e.g., containing) 1 to 30% G (guluronate) residues, 30 to 70% G (guluronate) residues, or 70 to 99% G (guluronate) residues. Thus, a typical MG block-containing alginate oligomer for use according to the present invention may contain more than 30% but less than 70% G residues (i.e., more than 30% but less than 70% of the monomer residues of the MG block alginate oligomer are G residues).

[0141] Preferably, more than 30%, more particularly more than 35%, or 40%, and even more particularly more than 45, 50, 55, 60, or 65% of the monomeric residues of the MG block-containing alginic acid oligomer are guluronate, but in each case less than 70%. Alternatively, less than 70%, more preferably less than 65%, or 60%, and even more preferably less than 55, 50, 45, 40, or 35% of the monomeric residues of the MG block-containing alginic acid oligomer are guluronate, but in each case more than 30%. Ranges formed by any combination of these values ​​may also be selected. Thus, for example, an MG block-containing alginic acid oligomer can have, for example, between 35% and 65%, 40% and 60%, or 45% and 55% of G residues.

[0142] In another embodiment, the MG block-containing alginate oligomer may have approximately equal amounts of G and M residues (e.g., 65%G / 35%M and 35%G / 65%M, e.g., 60%G / 40%M and 40%G / 60%M, 55%G / 45%M and 45%G / 55%M, 53%G / 47%M and 47%G / 53%M, 51%G / 49%M and 49%G / 51%M, e.g., ratios between about 50%G and about 50%M), with these residues arranged predominantly, preferably entirely, or as completely as possible, in an alternating MG pattern (e.g., at least 50%, or at least 60, 70, 80, 85, 90, or 95%, or 100% of the M and G residues are in the alternating MG sequence).

[0143] In certain embodiments, the terminal uronic acid residue of the oligomer of the invention does not have a double bond, particularly a double bond between the C4 and C5 atoms. Such oligomers may be described as having a saturated terminal uronic acid residue. That is, in other embodiments, the alginic acid oligomer of the invention has an unsaturated terminal uronic acid residue. Those skilled in the art will be able to prepare oligomers with saturated terminal uronic acid residues without undue burden. This may be by using manufacturing techniques that produce such oligomers, or by converting (saturating) oligomers produced by processes that produce oligomers with unsaturated terminal uronic acid residues.

[0144] As noted above, the monomer residues in the alginate oligomer can be the same or different, and need not all contain charged groups, although it is preferred that a majority (e.g., at least 60%, preferably at least 80%, more preferably at least 90%) contain charged groups. It is preferred that a substantial majority, e.g., at least 80%, more preferably at least 90%, of the charged groups have the same polarity. In the alginate oligomer, the ratio of hydroxyl groups to charged groups is preferably at least 2:1, especially at least 3:1.

[0145] Alginate oligomers are typically charged, and therefore counterions for alginate oligomers can be any physiologically acceptable ion, particularly those commonly used for charged drug substances, such as sodium, potassium, ammonium, chloride, mesylate, meglumine, etc. Ions that promote alginate gelation, such as Group 2 metal ions, can also be used. Others can include salt-forming ions, mentioned below in the context of therapeutic polypeptides.

[0146] Although alginate oligomers may be synthetic materials resulting from the polymerization of an appropriate number of guluronate and mannuronate residues, alginate oligomers for use in the present invention may be conveniently obtained, manufactured or derived from natural sources such as those mentioned above, i.e., natural alginate source materials.

[0147] Cleavage of polysaccharides into oligosaccharides to produce alginate oligomers usable according to the invention may be carried out using conventional polysaccharide dissolution techniques such as enzymatic digestion and acid hydrolysis. In one advantageous embodiment, acid hydrolysis is used to prepare the alginate oligomers of the invention. In another embodiment, enzymatic digestion is used with an additional processing step to saturate the terminal uronic acids in the oligomers.

[0148] The oligomers may then be separated from the polysaccharide degradation products chromatographically using ion exchange resins, or by fractional precipitation, solubilization, or filtration. U.S. Pat. No. 6,121,441 and WO 2008 / 125828, both of which are expressly incorporated herein by reference in their entireties, describe suitable processes for preparing alginate oligomers for use in the present invention. Further information and discussion can be found, for example, in "Handbooks of Hydrocolloids," edited by Phillips and Williams, CRC, Boca Raton, Florida, USA, 2000, which is expressly incorporated herein by reference in its entirety.

[0149] Alginate oligomers may also be chemically modified, including, but not limited to, modifications to add charged groups (e.g., carboxylated or carboxymethylated glycans) and alginate oligomers modified to alter flexibility (e.g., by periodate oxidation).

[0150] Alginate oligomers (e.g., oligoglucuronic acid) suitable for use according to the present invention may be conveniently prepared by acid hydrolysis of alginate, including but not limited to, from Laminaria hyperborea and Lessonia nigrescens, dissolving at neutral pH, lowering the pH to 3.4 by adding mineral acid to precipitate the alginate oligomers (oligoglucuronic acid), washing with weak acid, resuspension at neutral pH, and lyophilization.

[0151] Alginate for the preparation of the alginate oligomers of the present invention may also be obtained directly from a suitable bacterial source, such as Pseudomonas aeruginosa or Azotobacter vinelandii.

[0152] In embodiments where alginate oligomers are desired that have a primary structure in which the majority of G residues are arranged in G-blocks rather than as single residues, algal sources are expected to be most suitable due to the fact that alginates produced in these organisms tend to have these structures, whereas bacterial sources may be more suitable for obtaining alginate oligomers of various structures.

[0153] The molecular machinery involved in alginate biosynthesis in Pseudomonas fluorescens and Azotobacter vinelandii has been cloned and characterized (WO 94 / 09124; Ertesvaeg, H. et al., Metabolic Engineering, 1, 262-269 (1999); WO 2004 / 011628; Gimmestad, M. et al., op. cit.; Remminghorst and Rehm, Biotechnology Letters, 28, 1701-1712 (2006); Gimmestad, M. et al., Journal of Bacteriology, 188, No. 15, 5551-5560 (2006)).

[0154] The G content of alginate (e.g., algal source material) can be increased by epimerization using, for example, A. vinelandii mannuronan C-5 epimerase or other epimerase enzymes. Thus, for example, in vitro epimerization can be performed using epimerases isolated from Pseudomonas or Azotobacter, such as AlgG from Pseudomonas fluorescens or Azotobacter vinelandii, or AlgE enzymes (AlgE1-AlgE7) from Azotobacter vinelandii. The use of epimerases from other organisms capable of producing alginate, particularly algae, is also specifically contemplated. In vitro epimerization of low G alginate using Azotobacter vinelandii AlgE epimerase has been described in detail by Eltesberg et al. (supra) and Strugala et al. (Gums and Stabilisers for the Food Industry, 2004, Vol. 12, Royal Society of Chemistry, pp. 84-94).

[0155] To obtain G-block-containing alginate or alginate oligomers, epimerization using one or more Azotobacter vinelandii AlgE epimerases other than AlgE4 is preferred because these enzymes are capable of producing G-block structures. On the other hand, AlgE4 epimerase can be used to create alginate or alginate oligomers with primary structures containing alternating M / G strings or single G residues because this enzyme has been found to epimerize individual M residues to preferentially produce single G residues linked to M residues rather than G blocks. Specific primary structures can be obtained by using various combinations of these enzymes.

[0156] It is specifically contemplated that mutant versions of these enzymes or homologs from other organisms may also be used. WO 94 / 09124, for example, describes recombinant or modified mannuronan C-5 epimerase enzymes (AlgE enzymes) encoded by epimerase sequences in which DNA sequences encoding various regions or modules of the epimerase have been shuffled or deleted and recombined. Alternatively, variants of naturally occurring epimerase enzymes (AlgG or AlgE) may be used, for example, obtained by site-directed or random mutagenesis of the AlgG or AlgE gene.

[0157] Another approach is to create Pseudomonas and Azotobacter organisms in which some or all of the epimerase genes have been mutated in such a way that the mutants produce alginate of the required structure, or even alginate oligomers of the required structure and size (or molecular weight), for subsequent alginate oligomer production. International Publication No. 2004 / 011628 and Gimmestad, M. et al., 2003 (supra) detail the generation of several Pseudomonas fluorescens organisms with mutated AlgG genes. Gimmestad, M. et al., 2006 (supra) disclose the generation of several Azotobacter vinelandii organisms with mutated AlgE genes. Those skilled in the art will be able to use this teaching to generate new mutants that can be used to generate the alginate oligomers of the present invention without undue effort.

[0158] A further approach is to delete or inactivate the endogenous epimerase gene from an Azotobacter or Pseudomonas organism and then introduce one or more exogenous epimerase genes. The genes may be mutated or non-mutated (i.e., wild-type or modified), and their expression may be controlled, for example, using an inducible or other "controllable promoter." By selecting the appropriate combination of genes, alginate of a predetermined primary structure can be produced.

[0159] Another further approach would be to introduce some or all of the alginate biosynthetic machinery of Pseudomonas and / or Azotobacter into non-alginate producing organisms (e.g., E. coli) and genetically induce alginate production from these modified organisms.

[0160] When these culture-based systems are used, the culture conditions can influence the primary structure of the alginate or alginic acid oligomer product. Adjusting culture parameters such as temperature, osmolality, nutrient levels / sources, and atmospheric parameters to manipulate the primary structure of the alginate produced by a particular organism is well within the capabilities of one skilled in the art.

[0161] References to "G residue / G" and "M residue / M" or guluronic acid or mannuronic acid, or guluronate or mannuronate should be read interchangeably with references to guluronic acid / guluronate and mannuronic acid / mannuronate (specifically α-L-guluronic acid / guluronate and β-D-mannuronic acid / mannuronate), and further include derivatives thereof in which one or more available side chains or groups have been modified without resulting in a substantially less improvement than that of the unmodified oligomer in systemic bioavailability, gastric uptake, ability to increase absorption from the stomach, or efficacy of a therapeutic polypeptide in treating or preventing a disease or condition responsive to the therapeutic polypeptide or its complications. Common saccharide modifying groups include acetyl, sulfate, amino, deoxy, alcohol, aldehyde, ketone, ester, and anhydride groups. Any of these groups may be used to modify alginate oligomers according to the present invention. Alginate oligomers may also be chemically modified to add charged groups (e.g., carboxylated or carboxymethylated glycans) and to alter flexibility (e.g., by periodate oxidation). Those skilled in the art will recognize additional chemical modifications that can be made to the monosaccharide subunits of oligosaccharides and can apply these to the alginate oligomers of the present invention. In other embodiments, the alginate oligomer is unmodified, i.e., consists of unmodified G and / or M residues.

[0162] The present invention encompasses the use of a single alginate oligomer or a mixture of different alginate oligomer(s). Accordingly, the methods, uses, and compositions of the present invention may comprise at least one type, or one or more types of alginate oligomer. Thus, for example, combinations of different alginate oligomers (e.g., 2, 3, 4, 5, or more) may be used. In this regard, a combination of alginate oligomers may be selected that together provide a profile of properties advantageous for practicing the invention. This may be a combination of different oligomer sizes, different G and M contents, and / or different monomer configurations. References herein to "an" or "the" alginate oligomer should be construed to cover such embodiments unless otherwise indicated. Specifically, a composition described herein as "consisting of" a set of components of which "an alginate oligomer" is a part includes the combination of alginate oligomers as said components.

[0163] In certain embodiments, the alginate oligomer is not an alginate that acts as a gastrointestinal epithelial barrier (epithelial) permeability enhancer in the stomach of a subject, or in any part of the GI tract of a subject.

[0164] Therapeutic polypeptides, including the terms therapeutic proteins and peptides, polypeptide therapeutics, and polypeptide therapeutics and the like, can be any peptide or polypeptide capable of exerting a therapeutic effect in a subject, either directly or indirectly. Thus, a polypeptide can be an activator of a signal transduction pathway that results in a physiological effect, or can be a mediator of such a pathway by acting on an intermediate of such a pathway. A polypeptide can exert its effect by promoting or interfering with a physiological reaction or interaction between biomolecules in a subject, or can be an enzyme that can promote a physiological effect by its activity on cellular components of the subject or biomolecules therein. This is not an exhaustive list.

[0165] The term polypeptide, including peptide, oligopeptide, and protein, can refer to a polypeptide found naturally in a subject or other animal, or to a fragment, analog, and / or derivative thereof. Polypeptides can be artificial constructs containing naturally occurring amino acid sequences or fragments thereof, or artificial amino acid sequences. The amino acid units can be those found in naturally occurring proteins, or they can be artificial amino acids or modified forms of naturally occurring amino acids, e.g., D-forms. Polypeptides can be proteins with natural physiological functions, or they can contain amino acid sequences that have other biological effects when introduced into a physiological system, such as antigens. Antigens can be derived from parasitic or infectious species, optionally complexed with an immunogenic carrier. Such species can be, for example, bacteria, viruses, yeast, or fungi.

[0166] A therapeutic polypeptide can be of any size, but typically it is at least 500 Da, e.g., at least 750 Da, 1 kDa, 1.5 kDa, 2 kDa, 5 kDa, 10 kDa, 20 kDa, 50 kDa, or 100 kDa. In other embodiments, it is less than 2 MDa, e.g., less than 1.5 MDa, 1 MDa, 500 kDa, or 200 kDa. Any range having endpoints constructed from any of these values ​​is expressly contemplated.

[0167] Thus, therapeutic polypeptides include peptide hormones / growth factors (e.g., insulin, glucagon, glucagon-like peptide-1, somatostatin, angiotensin II, endothelin, gastrin, growth hormone, gonadotrophins, erythropoietin, colony-stimulating factors (e.g., G-CSF), VEGF, EGF, HGF, PDGF, FGF, neurotrophins (e.g., nerve growth factor (NGF), brain-derived neurotrophic factor (BDNF), neurotrophin-3, neurotrophin-4), cytokines (e.g., TNF, IL-1, IL-6, IL-8, IL-10, IFN-γ, IFN-α, IFN-β, IFN-ε, IFN-κ, and IFN-ω), blood factors (e.g., Factor VII, Factor VIII, and Factor IX), and enzymes (e.g., glucocerebrosidase, Cas 9, Cas 12, Cas 16, Cas 18, Cas 20, Cas 21, Cas 22, Cas 23, Cas 24, Cas 25, Cas 26, Cas 27, Cas 28, Cas 29, Cas 30, Cas 31, Cas 32, Cas 33, Cas 34, Cas 35, Cas 36, Cas 37, Cas 38, Cas 39, Cas 40, Cas 41, Cas 42, Cas 43, Cas 44, Cas 45, Cas 46, Cas 47, Cas 48, Cas 49, Cas 50, Cas 51, Cas 52, Cas 53, Cas 54, Cas 55, Cas 56, Cas 57, Cas 58, Cas 59, Cas 60, Cas 61, Cas 62, Cas 63, Cas 64, Cas 65, Cas 66, Cas 67 13) or their analogs. Amino acid sequences for such peptides are available in the technical literature.

[0168] Further specific examples include adrenocorticotropic hormone (ACTH), corticotropin-releasing factor, angiotensin, calcitonin, insulin, glucagon, glucagon-like peptide-1, glucagon-like peptide-2, insulin-like growth factor, insulin-like growth factor-2, gastric inhibitory peptide, growth hormone-releasing factor, pituitary adenylate cyclase-activating peptide, secretin, enterogastrone, somatostatin, somatotropin, somatomedin, parathyroid hormone, thrombopoietin, follicle-stimulating hormone, erythropoietin, hypothalamic-releasing factor, prolactin, thyroid-stimulating hormone, endorphins, enkephalins, vasopressin, oxytocin, interferons, and analogs thereof, superoxide dismutase, Cas9, Cas12, Cas13, asparaginase, arginase, arginine deaminase, adenosine deaminase, and ribonuclease. In certain embodiments, the therapeutic polypeptide is not angiotensin, calcitonin, insulin, insulin-like growth factor, or insulin-like growth factor-2.

[0169] In certain embodiments, the polypeptide therapeutic agent is not a lysozyme, a lysosome-derived polypeptide, or a lysosomal enzyme such as L-asparaginase.

[0170] In other embodiments, the polypeptide may be or include an immunoglobulin amino acid sequence, e.g., an amino acid sequence derived from IgG, IgA, IgM, IgE, or IgD. The immunoglobulin amino acid sequence may be a fragment, e.g., Fc, Fv, Fab, Fab2, scFV, and variants. In certain embodiments, the therapeutic polypeptide may be a therapeutic antibody, particularly a monoclonal antibody. Examples of therapeutic antibodies include alemtuzumab, bevacizumab, cetuximab, ofatumumab, panitumumab, rituximab, trastuzumab, ipilimumab, nivolumab, pembrolizumab, atezolizumab, avelumab, durvalumab, cemiplimab, stimulimab, anifrolumab, bimekizumab, trolokinumab, evinacumab, aducanumab, ansuvimab, atortivimab, teprotumumab, eptinezumab, crizanlizumab, bromococcal including ecizumab, risankizumab, romosozumab, galcanezumab, erenumab, ibalizumab, emicizumab, benralizumab, ocrelizumab, sarilumab, dupilumab, bezlotoxumab, ixekizumab, alirocumab, vedolizumab, tocilizumab, canakinumab, infliximab, adalimumab, omalizumab, efalizumab, golimumab, ustekinumab, certolizumab pegol, ibritumomab, or tositumomab.

[0171] In other embodiments, the polypeptide may be a polypeptide antibiotic, such as an anti-infective and anti-tumor antibiotic, including, for example, actinomycin, gramicidin, tylocidin, bleomycin, bacitracin, colistin, and polymyxin B.

[0172] In other embodiments, the polypeptide may be an antimicrobial peptide or protein, such as a host defense peptide or an innate immunity peptide. Such peptides are naturally occurring or synthetic peptides with antibacterial, antiviral, antifungal, and / or antiparasitic activity. Naturally occurring examples may be found in the Antimicrobial Peptide Database maintained by the University of Nebraska Medical Center (aps.unmc.edu / home).

[0173] The term "polypeptide therapeutic agent" and the like includes peptides bearing non-peptide chemical groups, such as carbohydrate, lipid, nucleic acid, or polyalcohol groups. In certain embodiments, the polypeptide may be an acylated or PEGylated polypeptide designed to have an increased plasma half-life, such as an acylated or PEGylated form of any of those disclosed above.

[0174] In further embodiments, the polypeptide may be a fusion protein or chimeric protein of two or more polypeptides or fragments thereof, any of the specific examples of which may be fused to, for example, albumin or Fc, or fragments thereof that increase plasma half-life.

[0175] In certain specific embodiments, the therapeutic polypeptide may be an incretin peptide or analog / mimetic thereof, such as glucagon, glucagon-like protein 1, gastric inhibitory peptide (glucose-dependent insulinotropic polypeptide (GIP) and exendin-4 peptides. Incretins are a group of metabolic hormones that stimulate a decrease in blood glucose levels. Incretins are released after a meal and increase the secretion of insulin from the pancreatic beta cells of the islets of Langerhans through a blood glucose-dependent mechanism. Such peptides or their analogs / mimetic may be considered agonists of the glucagon receptor, the glucose-dependent insulinotropic polypeptide (GIP) receptor and / or the glucagon-like peptide-1 (GLP-1) receptor. In certain embodiments, the therapeutic polypeptide may be a dual incretin peptidomimetic compound that is an agonist of the receptors for GIP and GLP-1. The therapeutic polypeptide may be a triple incretin peptidomimetic compound that is an agonist of the receptors for GIP, GLP-1 and glucagon. In more preferred embodiments, the polypeptide is an agonist of the human forms of the above receptors.

[0176] Thus, in certain specific embodiments, the therapeutic polypeptide may be dulaglutide (Trulicity), exenatide (Byetta), liraglutide (Victoza), semaglutide (Ozempic, Ugovy, and Ryvelsus), tirzepatide (Manjaro), albiglutide (Epelsan / Tanzeum), lixisenatide (Lyxumia / Adrixin), polyethylene glycol loxenatide (Flymay), and LY3437943 (Retatortide).

[0177] Similarly, in certain specific embodiments, the therapeutic polypeptide is selected from the group consisting of cotadutide, taspoglutide, langrenatide, veinaglutide, efpeglenatide, LY3502970 (Eli Lilly), LY3537031 (Eli Lilly), LY3493269 (Eli Lilly), HM12525A / JNJ-64565111 (efmoglutide, Hanmi Pharmaceutical, MOD6030 / 1, Prolor / OPKO Biologicals), SAR425899 (Sanofi), MEDI0382 (MedImmune), MK8521 (Merck), ZP2929 / BI456906 (Zealand-Boehringer), NN9709 / NNC0090-2746 / MAR709 / RG7697 / R06 These include 811135 (Novo Nordisk), SAR441255 (Sanofi), C2816 (MedImmune), ZP3022 (Zealand), NNC9204-1177 (NN9277) (Novo Nordisk), LY3305677 (Eli Lilly), JNJ-54728518 (Janssen), LY2944876 / TT-401 (Transition Therapeutics), CPD86 (Eli Lilly), SAR438335 (Sanofi), ZP-I-98 (Zealand), ZP-DI-70 (Zealand), HM15211 (Hanmi Pharmaceutical), NN9423 / MAR423 (Novo Nordisk / Marcadia), PB-719 (PegBio), and DD01 (D&D Pharma).

[0178] In certain specific embodiments, the therapeutic polypeptide may be an incretin analog disclosed in WO2019 / 125929 and WO2019125938, which are incorporated herein by reference.

[0179] In certain specific embodiments, the therapeutic polypeptide may be an oxyntomodulin peptide or an analog / mimetic thereof, such as mazdutide (LY3305677).

[0180] In certain specific embodiments, the therapeutic polypeptide may be any one of the polypeptide incretin analogs referenced in WO 2015 / 185640, WO 2015 / 086733, WO 2015 / 155139, WO 2013 / 164483 or WO 2015 / 086728, which are incorporated herein by reference.

[0181] In certain specific embodiments, the therapeutic polypeptide is selected from the group consisting of polypeptides described in WO 93 / 19175, WO 96 / 29342, WO 98 / 08871, WO 99 / 43707, WO 99 / 43706, WO 99 / 43341, WO 99 / 43708, WO 2005 / 027978, WO 2005 / 058954, WO 2005 / 05895, each of which is incorporated herein by reference. 8, WO2006 / 005667, WO2006 / 037810, WO2006 / 037811, WO2006 / 097537, WO2006 / 097538, WO2008 / 023050, WO2009 / 030738, WO2009 / 030771 or WO2009 / 030774.

[0182] A polypeptide therapeutic may be a fusion protein formed with any one of the polypeptides referred to herein, particularly an incretin analog.

[0183] In these embodiments, the methods of the invention may be aimed at treating any disease / disorder in which GLP-1, GIP or glucagon is implicated (including any disease / disorder in which a deficiency of GLP-1, GIP or glucagon or a deficiency of GLP-1, GIP or glucagon signaling is involved or mediated by). Thus, in these embodiments, the methods of the invention are directed to the treatment of excess weight, obesity, diabetes type II, insulin resistance, pre-diabetes, gestational diabetes, diabetes type I, hyperlipidemia, metabolic syndrome, cardiovascular disease (including atherosclerosis, myocardial infarction, coronary heart disease, stroke, cardiac insufficiency, heart failure (acute or chronic), coronary artery disease, cardiomyopathy, reperfusion injury, cerebral ischemia, left ventricular hypertrophy, arrhythmias, cardiac dysrhythmias, syncope, angina, stenosis or restenosis coronary artery disease), hypertension, fatty liver disease, non-alcoholic steatohepatitis (NASH), chronic kidney disease and polycystic ovary syndrome (PCOS), neurological dysfunction (e.g., dementia, Alzheimer's disease and Parkinson's disease), and the treatment of symptoms and complications associated with (attributed to / caused by) such disorders.

[0184] Common side effects observed with polypeptide incretin analogs can be nausea, vomiting, diarrhea, abdominal pain, and constipation. These effects may be reduced by the present invention, as it is expected that the present invention will allow for the use of lower doses.

[0185] GIP is a 42-amino acid gastrointestinal regulatory peptide that plays a physiological role in glucose homeostasis by stimulating insulin secretion from pancreatic β cells and protecting them in the presence of glucose. GLP-1 is a 37-amino acid peptide that stimulates insulin secretion, protects pancreatic β cells, and inhibits glucagon secretion, gastric emptying, and food intake, leading to weight loss. GIP and GLP-1 are known as incretins. Incretin receptor signaling exerts physiologically relevant actions important for glucose homeostasis. In normal physiology, GIP and GLP-1 are secreted from the intestine after a meal, and these incretins enhance physiological responses to food, including satiety, insulin secretion, and nutrient processing. Patients with type 2 disease (T2D) have reduced incretin responses.

[0186] Human GLP-1 is a 37-amino acid residue peptide derived from preproglucagon, which is synthesized, inter alia, in L cells in the distal ileum, the pancreas, and the brain. Human GLP-1(7-37) has the sequence HAEGTFTSDVSSYLEGQAAKEFIAWLVKGRG. Processing of preproglucagon to produce GLP-1(7-36)amide, GLP-1(7-37), and GLP-2 occurs primarily in L cells. A simple system is used to describe fragments and analogs of this peptide; e.g., Gly8-GLP-1(7-37) designates a fragment of GLP-1 formally derived from GLP-1 by deleting amino acid residues 1 through 6 and substituting Gly for the natural amino acid residue at position 8 (Ala). Similarly, Lys34(N) is replaced by Lys(N). ε(-tetradecanoyl)-GLP-1(7-37) designates GLP-1(7-37) in which the ε-amino group of the Lys residue at position 34 has been tetradecanoylated. When a C-terminally extended GLP1 analog is referred to herein, the amino acid residue at position 38 is Arg unless otherwise specified, the optional amino acid residue at position 39 is also Arg unless otherwise specified, and the optional amino acid residue at position 40 is Asp unless otherwise specified. Also, when a C-terminally extended analog extends to positions 41, 42, 43, 44, or 45, the amino acid sequence of the extension is the same as in the corresponding sequence in human preproglucagon unless otherwise specified. Similarly, in an N-terminally extended GLP1 analog, the amino acid sequence of the extension is the same as in the corresponding sequence in human preproglucagon.

[0187] As used herein, the term "GLP-1 agonist" refers to a compound that fully or partially activates the human GLP-1 receptor, and is therefore equivalent to the term "GLP-1 receptor agonist." In some embodiments, a "GLP-1 agonist" is ... D ) or binds to the GLP-1 receptor with a potency (EC ) of less than 1 μM, e.g., less than 100 nM, as measured by methods known in the art (see, e.g., WO 98 / 08871). 50 ) and exhibit insulinotropic activity. Here, insulinotropic activity may be measured in an in vivo or in vitro assay known to those skilled in the art. For example, a GLP-1 agonist may be administered to an animal with elevated blood glucose levels (e.g., obtained using an intravenous glucose tolerance test (IVGTT)), and a person skilled in the art can determine an appropriate glucose dose and an appropriate blood sampling schedule depending on, for example, the animal species for the IVGTT and the plasma insulin concentration values ​​measured over time.

[0188] The terms "GIP agonist" and "glucagon agonist" may be interpreted analogously.

[0189] In some embodiments, the GLP-1 agonist is a GLP-1 analog, optionally containing one substituent. The term "analog," as used herein with reference to a GLP-1 peptide (hereinafter "peptide"), refers to a peptide in which at least one amino acid residue of the peptide has been replaced with another amino acid residue, and / or at least one amino acid residue has been deleted from the peptide, and / or at least one amino acid residue has been added to the peptide, and / or at least one amino acid residue of the peptide has been modified. Such addition or deletion of amino acid residues may occur at the N-terminus of the peptide and / or at the C-terminus of the peptide.

[0190] GLP-1 analogs include fusion proteins containing one or more copies of GLP-1 or substitution variants thereof and one or more additional polypeptides, such as albumin, including albiglutide, a peptide consisting of 645 proteinogenic amino acids and 17 disulfide bridges linking amino acids 113-122, 135-151, 150-161, 184-229, 228-237, 260-306, 305-313, 325-339, 338-349, 376-421, 420-429, 452-498, 497-508, 521-537, 536-547, 574-619, 618-627. Amino acids 1-30 and 31-60 constitute two copies of modified GLP-1, with an alanine replacing a glycine at position 2, and the remaining sequence encoding human albumin.

[0191] In some embodiments, the GLP-1 agonist is a derivative of GLP-1, i.e., a GLP-1 analog. Herein, the parent peptide from which such a derivative is formally derived is referred to in some places as the "GLP-1 portion" of the derivative. The GLP-1 portion can be a native GLP-1 amino acid sequence, a fragment thereof, or an analog of such an amino acid sequence. The term "derivative" is used in the present context of GLP-1 peptides and analogs thereof to designate a peptide in which one or more of the amino acid residues of the parent peptide have been chemically modified, for example, by alkylation, acylation, ester formation, or amide formation.

[0192] In some embodiments, the GLP-1 agonist is a derivative of exendin-4, including lixisenatide, a C-terminal amide modified peptide consisting of the first 39 amino acids of the sequence of Gila monster (Heloderma suspectum) exendin-4, with the proline at position 38 omitted and six lysine residues added at the C-terminus.

[0193] In some embodiments, the GLP-1 agonist is a derivative of GIP. Human GIP has the sequence YAEGTFISDYSIAMDKIHQQDFVNWLLAQKGKKNDWKHNITQ.

[0194] One such example is the peptide LY3437943 (retatortide), which consists of 39 amino acids engineered from the amino acid sequence of human GIP, with three unencoded amino acids substituted (Aib (α-aminoisobutyric acid) at positions 2 and 20 and αMeL (α-methyl-L-leucine) at position 13), and a C20 aliphatic diacid moiety linked to a lysine residue at position 17 via an AEE AγGlu linker (Coskun et al., Clinical and Translational Reports, Vol. 34, No. 9, pp. 1234-1247; WO 2019125938).

[0195] In certain specific embodiments, the therapeutic polypeptide may be liraglutide and structurally related analogs thereof. Liraglutide has the structure of Formula I, as shown in Figure 16A.

[0196] [ka]

[0197] In certain embodiments, the therapeutic polypeptide may be a GLP-1 derivative as described in WO 98 / 08871, e.g., a GLP-1 derivative to which a lipophilic substituent is attached, provided that at least one amino acid residue of the parent peptide has only one lipophilic substituent present and that, if this substituent is attached to an amino acid residue at the N-terminus or C-terminus of the parent peptide, this substituent is an alkyl group or a group bearing an ω-carboxylic acid group.

[0198] In another embodiment, the therapeutic polypeptide may be a GLP-1 derivative having only one lipophilic substituent.

[0199] In another embodiment, the therapeutic polypeptide may be a GLP-1 derivative having only one lipophilic substituent, which is an alkyl group or a group having an ω-carboxylic acid group, attached to the N-terminal amino acid residue of the parent peptide.

[0200] In another embodiment, the therapeutic polypeptide may be a GLP-1 derivative having only one lipophilic substituent, which is an alkyl group or a group having an ω-carboxylic acid group, attached to the C-terminal amino acid residue of the parent peptide.

[0201] In another embodiment, the therapeutic polypeptide may be a GLP-1 derivative having only one lipophilic substituent, which may be attached to any one amino acid residue that is neither the N-terminal nor the C-terminal amino acid residue of the parent peptide.

[0202] In another embodiment, the therapeutic polypeptide may be a GLP-1 derivative in which two lipophilic substituents are present.

[0203] In another embodiment, the therapeutic polypeptide may be a GLP-1 derivative in which two lipophilic substituents are present, one attached to the N-terminal amino acid residue and the other attached to the C-terminal amino acid residue.

[0204] In another embodiment, the therapeutic polypeptide may be a GLP-1 derivative in which two lipophilic substituents are present, one attached to the N-terminal amino acid residue and the other attached to an amino acid residue that is not the N-terminal or C-terminal amino acid residue.

[0205] In another embodiment, the therapeutic polypeptide may be a GLP-1 derivative in which two lipophilic substituents are present, one attached to the C-terminal amino acid residue and the other attached to an amino acid residue that is not the N-terminal or C-terminal amino acid residue.

[0206] In a further embodiment, the therapeutic polypeptide may be a derivative of GLP-1(7-C), where C is selected from the group consisting of 38, 39, 40, 41, 42, 43, 44 and 45, which derivative has only one lipophilic substituent attached to the C-terminal amino acid residue of the parent peptide.

[0207] In a further embodiment, the therapeutic polypeptide may be a GLP-1 derivative in which the lipophilic substituent comprises from 4 to 40 carbon atoms, for example from 8 to 25 carbon atoms.

[0208] In a further embodiment, the therapeutic polypeptide may be a GLP-1 derivative in which a lipophilic substituent is attached to an amino acid residue in such a way that the carboxyl group of the lipophilic substituent forms an amide bond with the amino group of the amino acid residue.

[0209] In a further embodiment, the therapeutic polypeptide may be a GLP-1 derivative in which a lipophilic substituent is attached to an amino acid residue in such a way that the amino group of the lipophilic substituent forms an amide bond with the carboxyl group of the amino acid residue.

[0210] In a further embodiment, the therapeutic polypeptide may be a GLP-1 derivative in which a lipophilic substituent is attached to the parent peptide using a spacer.

[0211] In a further embodiment, the therapeutic polypeptide may be a GLP-1 derivative in which a lipophilic substituent is attached, optionally via a spacer, to the ε-amino group of a Lys residue contained in the parent peptide.

[0212] In a further embodiment, the therapeutic polypeptide may be a GLP-1 derivative in which the lipophilic substituent is attached to the parent peptide using a spacer which is an unbranched alkane α,ω-dicarboxylic acid group having one to seven methylene groups, preferably two methylene groups, and which spacer forms a bridge between the amino group of the parent peptide and the amino group of the lipophilic substituent.

[0213] In further embodiments, the therapeutic polypeptide may be a GLP-1 derivative in which a lipophilic substituent is attached to the parent peptide using a spacer that is an amino acid residue other than Cys or a dipeptide such as Gly-Lys. As used herein, the expression "dipeptide such as Gly-Lys" is used to designate a dipeptide in which the C-terminal amino acid residue is Lys, His, or Trp, preferably Lys, and the N-terminal amino acid residue is selected from the group consisting of Ala, Arg, Asp, Asn, Gly, Glu, Gln, Ile, Leu, Val, Phe, and Pro.

[0214] In a further embodiment, the therapeutic polypeptide may be a GLP-1 derivative in which a lipophilic substituent is attached to a parent peptide using a spacer which is an amino acid residue other than Cys or a dipeptide such as Gly-Lys, and in which the carboxyl group of the parent peptide forms an amide bond with the amino group of the Lys residue or dipeptide containing a Lys residue, and the other amino group of the Lys residue or dipeptide containing a Lys residue forms an amide bond with the carboxyl group of the lipophilic substituent.

[0215] In further embodiments, the therapeutic polypeptide may be a GLP-1 derivative in which a lipophilic substituent is attached to a parent peptide using a spacer that is an amino acid residue other than Cys or a dipeptide such as Gly-Lys, and in which the amino group of the parent peptide forms an amide bond with the carboxyl group of the amino acid residue or dipeptide spacer, and the amino group of the amino acid residue or dipeptide spacer forms an amide bond with the carboxyl group of the lipophilic substituent.

[0216] In further embodiments, the therapeutic polypeptide may be a GLP-1 derivative in which the lipophilic substituent is attached to the parent peptide by a spacer which is an amino acid residue other than Cys or a dipeptide such as Gly-Lys, and in which the carboxyl group of the parent peptide forms an amide bond with the amino group of the amino acid residue spacer or dipeptide spacer, and the carboxyl group of the amino acid residue spacer or dipeptide spacer forms an amide bond with the amino group of the lipophilic substituent.

[0217] In a further embodiment, the therapeutic polypeptide may be a GLP-1 derivative in which the lipophilic substituent is attached to the parent peptide using a spacer which is an amino acid residue other than Cys or a dipeptide such as Gly-Lys, and the carboxyl group of the parent peptide forms an amide bond with the amino group of the spacer which is Asp or Glu or a dipeptide spacer containing an Asp or Glu residue, and the carboxyl group of the spacer forms an amide bond with the amino group of the lipophilic substituent.

[0218] In a further embodiment, the therapeutic polypeptide may be a GLP-1 derivative having a lipophilic substituent comprising a partially or fully hydrogenated cyclopentanophenanthrene backbone.

[0219] In a further embodiment, the therapeutic polypeptide may be a GLP-1 derivative having a lipophilic substituent that is a straight-chain or branched alkyl group.

[0220] In a further embodiment, the therapeutic polypeptide may be a GLP-1 derivative having a lipophilic substituent that is an acyl group of a straight-chain or branched fatty acid.

[0221] In a further embodiment, the therapeutic polypeptide comprises a CH3(CH2) n CO-, wherein n is an integer from 4 to 38, preferably an integer from 4 to 24, such as CH3(CH2)6CO-, CH3(CH2)8CO-, CH3(CH2) 10 CO-, CH3(CH2) 12 CO-, CH3(CH2) 14 CO-, CH3(CH2) 16 CO-, CH3(CH2) 18 CO-, CH3(CH2) 20 CO- and CH3(CH2) 22C There may be GLP-1 derivatives having a lipophilic substituent that is an acyl group selected from the group consisting of O-.

[0222] In a further embodiment, the therapeutic polypeptide may be a GLP-1 derivative having a lipophilic substituent that is an acyl group of a straight-chain or branched alkane α,ω-dicarboxylic acid.

[0223] In a further embodiment, the therapeutic polypeptide is HOOC(CH2) m CO-, where m is an integer from 4 to 38, preferably an integer from 4 to 24, such as HOOC(CH2) 12 CO-, HOOC(CH2) 14 CO-, HOOC(CH2)16 CO-, HOOC(CH2) 18 CO-, HOOC(CH2) 20 CO- and HOOC(CH2) 22 There may be GLP-1 derivatives having a lipophilic substituent that is an acyl group selected from the group consisting of CO-.

[0224] In a further embodiment, the therapeutic polypeptide has the formula CH3(CH2) p ((CH2) q In some embodiments, the GLP-1 derivative has a lipophilic substituent which is a group of formula: COOH)CHNHCO(CH2)2CO-, where p and q are integers and p+q is an integer from 8 to 33, preferably 12 to 28.

[0225] In a further embodiment, the therapeutic polypeptide has the formula CH3(CH2) r There may be GLP-1 derivatives having a lipophilic substituent that is a group of the formula CONHCH(COOH)(CH2)2CO-, where r is an integer from 10 to 24.

[0226] In a further embodiment, the therapeutic polypeptide has the formula CH3(CH2) s There may be GLP-1 derivatives having a lipophilic substituent that is a group of the formula CONHCH((CH2)2COOH)CO-, where s is an integer from 8 to 24.

[0227] In a further embodiment, the therapeutic polypeptide has the formula COOH(CH) t There may be GLP-1 derivatives having a lipophilic substituent that is a group of the formula: CO-, where t is an integer from 8 to 24.

[0228] In a further embodiment, the therapeutic polypeptide has the formula -NHCH(COOH)(CH)NHCO(CH) u The GLP-1 derivative may have a lipophilic substituent that is a group of the formula CH3, where u is an integer from 8 to 18.

[0229] In a further embodiment, the therapeutic polypeptide has the formula -NHCH(COOH)(CH2)4NHCOCH((CH2)2COOH)NH-CO(CH2) w The GLP-1 derivative may have a lipophilic substituent that is a group of formula CH3, where w is an integer from 10 to 16.

[0230] In a further embodiment, the therapeutic polypeptide has the formula -NHCH(COOH)(CH2)4NHCO(CH2)2CH(COOH)NHCO(CH2) X The GLP-1 derivative may have a lipophilic substituent that is a group of formula CH3, where x is an integer from 10 to 16.

[0231] In a further embodiment, the therapeutic polypeptide has the formula -NHCH(COOH)(CH2)4NHCO(CH2)2CH(COOH)NHCO(CH2) y The GLP-1 derivative may have a lipophilic substituent that is a group of the formula CH3, where y is 0 or an integer from 1 to 22.

[0232] In a further embodiment, the therapeutic polypeptide may be a GLP-1 derivative having a lipophilic substituent that can be negatively charged. Such a lipophilic substituent may be, for example, a substituent having a carboxyl group.

[0233] In a further embodiment, the therapeutic polypeptide may be a GLP-1 derivative, where the parent peptide is selected from the group consisting of GLP-1(1-45) or an analog thereof.

[0234] In a further embodiment, the therapeutic polypeptide may be a GLP-1 derivative or analogue thereof derived from a GLP-1 fragment selected from the group consisting of GLP-1(7-35), GLP-1(7-36), GLP-1(7-36)amide, GLP-1(7-37), GLP-1(7-38), GLP-1(7-39), GLP-1(7-40) and GLP1(7-41).

[0235] In further embodiments, the therapeutic polypeptide may be a GLP-1 analog or analog thereof derived from a GLP-1 analog selected from the group consisting of GLP-1(1-35), GLP-1(1-36), GLP-1(1-36)amide, GLP-1(1-37), GLP-1(1-38), GLP-1(1-39), GLP-1(1-40) and GLP-1(1-41).

[0236] In a further embodiment, the therapeutic polypeptide may be a GLP-1 derivative, where the designated analog comprises a derivative in which a total of up to 15, preferably up to 10, amino acid residues are replaced with any α-amino acid residue.

[0237] In a further embodiment, the therapeutic polypeptide may be a GLP-1 derivative, where the designated analog comprises a derivative in which a total of up to 15, preferably up to 10, amino acid residues are replaced with any α-amino acid residue that can be encoded by the genetic code.

[0238] In further embodiments, the therapeutic polypeptide may be a GLP-1 derivative, including derivatives in which a total of up to six amino acid residues have been replaced with alternative α-amino acid residues that can be encoded by the genetic code.

[0239] In a further embodiment, the therapeutic polypeptide may be a GLP-1(AB) derivative, where A is an integer from 1 to 7 and B is an integer from 38 to 45, or an analog thereof, which comprises one lipophilic substituent attached to the C-terminal amino acid residue and, optionally, a second lipophilic substituent attached to one of the other amino acid residues.

[0240] In a further embodiment, the parent peptide for the GLP-1 derivative of use in the present invention is Arg26-GLP-1(7-37), Arg34-GLP-1(7-37), Lys36-GLP-1(7-37), Arg26,34Lys36-GLP-1(7-37), Arg26,34Lys38GLP-1(7-38), Arg26,34Lys39-GLP-1(7-39) , Arg26, 34Lys40-GLP-1(7-40), Arg26Lys36-GLP-1(7-37), Arg34Lys36-GLP-1(7-37), Arg26Lys39-GL P1(7-39), Arg34Lys40-GLP-1(7-40), Arg26, 34Lys36, 39-GLP-1(7-39), Arg26, 34Lys36, 40-GLP-1(7-4 0), Gly8Arg26-GLP-1(7-37), Gly8Arg34-GLP-1(7-37), Gly8Lys36-GLP-1(7-37), Gly8Arg26, 34Lys36 -GLP-1(7-37), Gly8Arg26, 34Lys39-GLP-1(7-39), Gly8Arg26, 34Lys40-GLP-1(7-40), Gly8Arg26Lys3 and Gly8Arg26,34Lys36,40-GLP-1(7-40).

[0241] In a further embodiment, the parent peptide for the GLP-1 derivative for use in the present invention is Arg26, 34Lys38GLP-1(7-38), Arg26, 34Lys39GLP-1(7-39), Arg26, 34Lys40GLP-1(7-40), Arg26, 34Lys41GLP-1(7-41), Arg26, 34Lys42GLP-1(7-42), Arg26, 34Lys43GLP-1(7-43), Arg26, 34Lys44GLP-1(7-44), Arg26, 34Lys45GLP-1(7-45), Arg26 , 34Lys38GLP-1(1-38), Arg26, 34Lys39GLP-1(1-39), Arg26, 34Lys40GLP-1(1-40), Arg26, 34Lys41GLP-1(1-41), Arg26, 34Lys42GLP-1(1-42), Arg26 , 34Lys43GLP-1(1-43), Arg26, 34Lys44GLP-1(1-44), Arg26, 34Lys45GLP-1(1-45), Arg26, 34Lys38GLP-1(2-38), Arg26, 34Lys39GLP-1(2-39), Arg26 , 34Lys40GLP-1(2-40), Arg26, 34Lys41GLP-1(2-41), Arg26, 34Lys42GLP-1(2-42), Arg26, 34Lys43GLP-1(2-43), Arg26, 34Lys44GLP-1(2-44), Arg26 , 34Lys45GLP-1(2-45), Arg26, 34Lys38GLP-1(3-38), Arg26, 34Lys39GLP-1(3-39), Arg26, 34Lys40GLP-1(3-40), Arg26, 34Lys41GLP-1(3-41), Arg26 , 34Lys42GLP-1(3-42), Arg26, 34Lys43GLP-1(3-43), Arg26, 34Lys44GLP-1(3-44), Arg26, 34Lys45GLP-1(3-45), Arg26, 34Lys38GLP-1(4-38), Arg26 , 34Lys39GLP-1(4-39), Arg26, 34Lys40GLP-1(4-40), Arg26, 34Lys41GLP-1 (4-41), Arg26, 34Lys42GLP-1(4-42), Arg26, 34Lys43GLP-1(4-43), Arg26,34Lys44GLP-1(4-44), Arg26, 34Lys45GLP-1(4-45), Arg26, 34Lys38GLP-1(5-38), Arg26, 34Lys39GLP-1(5-39), Ar g26, 34Lys40GLP-1(5-40), Arg26, 34Lys41GLP-1(5-41), Arg26, 34Lys42GLP-1(5-42), Arg26, 34Lys43GLP-1(5-43 ), Arg26, 34Lys44GLP-1(5-44), Arg26, 34Lys45GLP-1(5-45), Arg26, 34Lys38GLP-1(6-38), Arg26, 34Lys39GLP-1( 6-39), Arg26, 34Lys40GLP-1(6-40), Arg26, 34Lys41GLP-1(6-41), Arg26, 34Lys42GLP-1(6-42), Arg26, 34Lys43GL P-1(6-43), Arg26, 34Lys44GLP-1(6-44), Arg26, 34Lys45GLP-1(6-45), Arg26Lys38GLP1(1-38), Arg34Lys38GLP-1 (1-38), Arg26, 34Lys36, 38GLP-1(1-38), Arg26Lys38GLP-1(7-38), Arg34Lys38GLP-1(7-38), Arg26, 34Lys36, 38G Selected from the group consisting of LP-1(7-38), Arg26,34Lys38GLP-1(7-38), Arg26Lys39GLP1(1-39), Arg34Lys39GLP-1(1-39), Arg26,34Lys36,39GLP-1(1-39), Arg26Lys39GLP-1(7-39), Arg34Lys39GLP-1(7-39) and Arg26,34Lys36,39GLP-1(7-39).

[0242] In a further embodiment, the therapeutic polypeptide is selected from the group consisting of the parent peptides Arg26-GLP-1(7-37), Arg34-GLP-1(7-37), Lys36-GLP-1(7-37), Arg26,34Lys36-GLP-1(7-37), Arg26Lys36-GLP-1(7-37), Arg34Lys36-GLP-1(7-37), Gly6Arg26 -GLP-1(7-37), Gly8Arg34-GLP-1(7-37), Gly8Lys36-GLP-1(7-37), Gly8Arg26,34Lys-GLP-1(7-37), Gly8Arg26Lys36-GLP-1(7-37) and Gly8Arg34Lys36-GLP-1(7-37).

[0243] In further embodiments, the therapeutic polypeptide may be a GLP-1 derivative wherein the parent peptide is selected from the group consisting of Arg26Lys38-GLP-1(7-38), Arg26,34Lys38-GLP-1(7-38), Arg26,34Lys36,38-GLP-1(7-38), Gly8Arg26Lys38-GLP-1(7-38) and Gly8Arg26,34Lys36,38-GLP-1(7-38).

[0244] In further embodiments, the therapeutic polypeptide may be a GLP-1 derivative wherein the parent peptide is selected from the group consisting of Arg26Lys39-GLP-1(7-39), Arg26,34Lys36,39-GLP-1(7-39), Gly8Arg26Lys39-GLP-1(7-39) and Gly8Arg26,34Lys36,39-GLP-1(7-39).

[0245] In further embodiments, the therapeutic polypeptide may be a GLP-1 derivative wherein the parent peptide is selected from the group consisting of Arg34Lys40-GLP-1(7-40), Arg26,34Lys36,40-GLP-1(7-40), Gly8Arg34Lys40-GLP-1(7-40) and Gly8Arg26,34Lys36,40-GLP-1(7-40).

[0246] In a further embodiment, the therapeutic polypeptide is Lys26(N ε -tetradecanoyl)-GLP-1(7-37), Lys34(N ε -tetradecanoyl)-GLP-1(7-37), Lys26,34-bis(N ε -tetradecanoyl)-GLP-1(7-37), Gly8Lys26(N ε -tetradecanoyl)-GLP-1(7-37), Gly8Lys34(N ε -tetradecanoyl)-GLP-1(7-37), Gly8Lys26,34-bis(N ε -tetradecanoyl)-GLP-1(7-37), Arg26Lys34(N ε -tetradecanoyl)-GLP-1(7-37), Lys26(N ε -tetradecanoyl)-GLP-1(7-38), Lys34(N ε -tetradecanoyl)-GLP-1(7-38), Lys26,34-bis(N ε -tetradecanoyl)-GLP-1(7-38), Gly8Lys26(N ε -tetradecanoyl)-GLP-1(7-38), Gly8Lys34(N ε -tetradecanoyl)-GLP-1(7-38), Gly8Lys26,34-bis(N ε -tetradecanoyl)-GLP-1(7-38), Arg26Lys34(N ε -tetradecanoyl)-GLP-1(7-38), Lys26(N ε -tetradecanoyl)-GLP-1(7-39), Lys34(N ε-tetradecanoyl)-GLP-1(7-39), Lys26,34-bis(N ε -tetradecanoyl)-GLP-1(7-39), Gly8Lys26(N ε -tetradecanoyl)-GLP-1(7-39), GlyaLys34(N ε -tetradecanoyl)-GLP-1(7-39), Gly8Lys26,34-bis(N ε -tetradecanoyl)-GLP-1(7-39), Arg26Lys34(N ε -tetradecanoyl)-GLP-1(7-39), Lys26(N ε -tetradecanoyl)-GLP-1(7-40), Lys34(N ε -tetradecanoyl)-GLP-1(7-40), Lys26,34-bis(N ε -tetradecanoyl)-GLP-1(7-40), Gly8Lys26(N ε -tetradecanoyl)-GLP-1(7-40), Gly8Lys34(N ε -tetradecanoyl)-GLP-1(7-40), Gly8Lys26,34-bis(N ε -tetradecanoyl)-GLP-1(7-40), Arg26Lys34(N ε -tetradecanoyl)-GLP-1(7-40), Lys26(N ε -tetradecanoyl)-GLP-1(7-36), Lys34(N ε -tetradecanoyl)-GLP-1(7-36), Lys26,34-bis(N ε -tetradecanoyl)-GLP-1(7-36), Gly8Lys26(N ε -tetradecanoyl)-GLP-1(7-36), Gly8Lys34(N ε -tetradecanoyl)-GLP-1(7-36), Gly8Lys26,34-bis(N ε -tetradecanoyl)-GLP-1(7-36), Arg26Lys34(N ε -tetradecanoyl)-GLP-1(7-36), Lys26(N ε -tetradecanoyl)-GLP-1(7-35), Lys34(N ε -tetradecanoyl)-GLP-1(7-35), Lys26,34-bis(N ε -tetradecanoyl)-GLP-1(7-35), Gly8Lys26(N ε -tetradecanoyl)-GLP-1(7-35), Gly8Lys34(N ε -tetradecanoyl)-GLP-1(7-35), Gly8Lys26,34-bis(N ε -tetradecanoyl)-GLP-1(7-35), Arg26Lys34(N ε -tetradecanoyl)-GLP-1(7-35), Lys26(N ε -tetradecanoyl)-GLP-1(7-36)amide, Lys34(N ε -tetradecanoyl)-GLP-1(7-36)amide, Lys26,34-bis(N ε -tetradecanoyl)-GLP-1(7-36)amide, Gly8Lys26(N ε -tetradecanoyl)-GLP-1(7-36)amide, Gly8Lys34(N ε -tetradecanoyl)-GLP-1(7-36)amide, Gly8Lys26,34-bis(N ε -tetradecanoyl)-GLP-1(7-36)amide, Arg26Lys34(Nε -tetradecanoyl)-GLP-1(7-36)amide, Gly8Arg26Lys34(N ε -tetradecanoyl)-GLP-1(7-37), Lys26(N ε -tetradecanoyl)Arg34-GLP-1(7-37), Gly8Lys26(N ε -tetradecanoyl)Arg34-GLP-1(7-37), Arg26,34Lys36(N ε -tetradecanoyl)-GLP-1(7-37), Gly8Arg26,34Lys36(N ε -tetradecanoyl)-GLP-1(7-37), Gly8Arg26Lys34(N ε -tetradecanoyl)-GLP-1(7-38), Lys26(N ε -tetradecanoyl)Arg34-GLP-1(7-38), Gly8Lys26(N ε -tetradecanoyl)Arg34-GLP-1(7-38), Arg26,34Lys36(N ε -tetradecanoyl)-GLP-1(7-38), Arg26,34 Lys38(N ε -tetradecanoyl)-GLP-1(7-38), Gly8Arg26,34Lys36(N ε -tetradecanoyl)-GLP-1(7-38), Gly8Arg26Lys34(N ε -tetradecanoyl)-GLP-1(7-39), Lys26(N ε -tetradecanoyl)Arg34-GLP-1(7-39), Gly8Lys26(N ε -tetradecanoyl)Arg34-GLP-1(7-39), Arg26,34Lys(N ε -tetradecanoyl)-GLP-1(7-39), Gly8Arg26,34Lys36(N ε -tetradecanoyl)-GLP-1(7-39), Gly8Arg26LysY(N ε -tetradecanoyl)-GLP-1(7-40), Lys26(N ε -tetradecanoyl)Arg34-GLP-1(7-40), Gly8Lys26(N ε -tetradecanoyl)Arg34-GLP-1(7-40), Arg26,34Lys36(N ε -tetradecanoyl)-GLP-1(7-40), Gly8Arg26.34(N ε -tetradecanoyl)-GLP-1(7-40), Lys26(N ε -(ω-carboxynonadecanoyl)-GLP-1(7-37), Lys34(N ε -(ω-carboxynonadecanoyl)-GLP-1(7-37), Lys26,34-bis(N ε -(ω-carboxynonadecanoyl)-GLP-1(7-37), Gly8Lys26(N ε -(ω-carboxynonadecanoyl)-GLP-1(7-37), Gly8Lys34(N ε -(ω-carboxynonadecanoyl)-GLP-1(7-37), Gyl8Lys26.34-bis(N ε -(ω-carboxynonadecanoyl)-GLP-1(7-37), Lys26(N ε -(ω-carboxynonadecanoyl)-GLP-1(7-38), Lys34(N ε -(ω-carboxynonadecanoyl)-GLP-1(7-38), Lys26,34-bis(N ε -(ω-carboxynonadecanoyl)-GLP-1(7-38), Gly8Lys26(N ε -(ω-carboxynonadecanoyl)-GLP-1(7-38), Gly2Lys34(N ε -(ω-carboxynonadecanoyl)-GLP-1(7-38), Gly8Lys26,34-bis(N ε -(ω-carboxynonadecanoyl)-GLP-1(7-38), Lys26(N ε -(ω-carboxynonadecanoyl)-GLP-1(7-39), Lys34(N ε -(ω-carboxynonadecanoyl)-GLP-1(7-39), Lys26,34-bis(N ε -(ω-carboxynonadecanoyl)-GLP-1(7-39), Gly8Lys26(N ε -(ω-carboxynonadecanoyl)-GLP-1(7-39), Gly8Lys34(N ε -(ω-carboxynonadecanoyl)-GLP-1(7-39), Gly8Lys26,34-bis(N ε -(ω-carboxynonadecanoyl)-GLP-1(7-39), Lys26(N ε -(ω-carboxynonadecanoyl)-GLP-1(7-40), Lys34(N ε -(ω-carboxynonadecanoyl)-GLP-1(740), Lys26,34-bis(N ε -(ω-carboxynonadecanoyl)-GLP-1(7-40), Gly8Lys26(N ε -(ω-carboxynonadecanoyl)-GLP-1(7-40), Gly8Lys34(N ε -(ω-carboxynonadecanoyl)-GLP-1(7-40), Gly8Lys26.34(N ε-(ω-carboxynonadecanoyl)-GLP-1(7-40), Lys26(N ε -(ω-carboxynonadecanoyl)-GLP-1(7-36), Lys34(N ε -(ω-carboxynonadecanoyl)-GLP-1(7-36), Lys26,34-bis(N ε -(ω-carboxynonadecanoyl)-GLP-1(7-36), Gly8Lys26(N ε -(ω-carboxynonadecanoyl)-GLP-1(7-36), Gly8Lys34(N ε -(ω-carboxynonadecanoyl)-GLP-1(7-36), Gly8Lys26,34-bis(N ε -(ω-carboxynonadecanoyl)-GLP-1(7-36), Lys26(N ε -(w-carboxynonadecanoyl)-GLP-1(7-36)amide, Lys34(N ε -(ω-carboxynonadecanoyl)-GLP-1(7-36)amide, Lys26,34-bis(N ε -(ω-carboxynonadecanoyl)-GLP-1(7-36)amide, Gly8Lys26(N ε -(ω-carboxynonadecanoyl)-GLP-1(7-36)amide, Gly8Lys(N ε -(ω-carboxynonadecanoyl)-GLP-1(7-36)amide, Gyl8Lys26,34-bis(N ε -(ω-carboxynonadecanoyl)-GLP-1(7-36)amide, Lys26(N ε -(ω-carboxynonadecanoyl)-GLP-1(7-35), Lys34(N ε -(ω-carboxynonadecanoyl)-GLP-1(7-35), Lys26,34-bis(N ε -(ω-carboxynonadecanoyl)-GLP-1(7-35), Gly8Lys26(N ε -(ω-carboxynonadecanoyl)-GLP-1(7-35), Gly8Lys34(N ε -(ω-carboxynonadecanoyl)-GLP-1(7-35), Gly8Lys26,34-bis(N ε -(ω-carboxynonadecanoyl)-GLP-1(7-35), Arg26Lys34(N ε -(ω-carboxynonadecanoyl)-GLP-1(7-37), Gly8Arg26Lys34(N ε -(ω-carboxynonadecanoyl)-GLP-1(7-37), Lys26(N ε -(ω-carboxynonadecanoyl))Arg34-GLP-1(7-37), Gly8Lys26(N ε -(ω-carboxynonadecanoyl))Arg34-GLP-1(7-37), Arg26,34Lys36(N ε -(ω-carboxynonadecanoyl)-GLP-1(7-37), Gly8Arg26,34Lys36(N ε -(ω-carboxynonadecanoyl)-GLP-1(7-37), Arg26Lys34(N ε -(ω-carboxynonadecanoyl)-GLP-1(7-38), Gly8Arg26Lys34(N ε -(ω-carboxynonadecanoyl)-GLP-1(7-38), Lys26(N ε -(ω-carboxynonadecanoyl))Arg34-GLP-1(7-38), Gly8Lys26(N ε -(O-carboxynonadecanoyl))Arg34-GLP-1(7-38), Arg26,34Lys36(N ε -(ω-carboxynonadecanoyl)-GLP-1(7-38), Arg26,34Lys36(N ε -(ω-carboxynonadecanoyl)-GLP-1(7-38), Gly8Arg26,34Lys36(N ε -(ω-carboxynonadecanoyl)-GLP-1(7-38), Arg26Lys34(N ε -(ω-carboxynonadecanoyl)-GLP-1(7-39), Gly8Arg26Lys34(N ε -(ω-carboxynonadecanoyl)-GLP-1(7-39), Lys26(N ε -(ω-carboxynonadecanoyl))Arg34-GLP-1(7-39), Gly8Lys26(N ε -(ω-carboxynonadecanoyl))Arg34-GLP-1(7-39), Arg26,34Lys36(N ε -(ω-carboxynonadecanoyl)-GLP-1(7-39), Gly8Arg26.34Lys38(N ε -(ω-carboxynonadecanoyl)-GLP-1(7-39), Arg26Lys34(N ε -(ω-carboxynonadecanoyl)-GLP-1(7-40), Gly8Arg26Lys34(N ε -(ω-carboxynonadecanoyl)-GLP-1(7-40), Lys28(N ε -(ω-carboxynonadecanoyl))Arg34-GLP-1(7-40), Gly8Lys26(N ε -(ω-carboxynonadecanoyl))Arg34-GLP-1(7-40), Arg26,34Lys36(N ε-(ω-carboxynonadecanoyl)-GLP-1(7-40), Gly8Arg26,34Lys36(N ε -(ω-carboxynonadecanoyl)-GLP-1(7-40), Lys26(N ε -(7-deoxycholoyl)-GLP-1(7-37), Lys34(N ε -(7-deoxycholoyl)-GLP-1(7-37), Lys26,34-bis(N ε -(7-deoxycholoyl)-GLP-1(7-37), Gly8Lys26(N ε -(7-deoxycholoyl)-GLP-1(7-37), Gly8Lys34(N ε -(7-deoxycholoyl)-GLP-1(7-37), Gly8Lys26,34-bis(N ε -(7-deoxycholoyl)-GLP-1(7-37), Arg26Lys34(N ε -(7-deoxycholoyl)-GLP-1(7-37), Lys26(N ε -(7-deoxycholoyl)-GLP-1(7-38), Lys34(N ε -(7-deoxycholoyl)-GLP-1(7-38), Lys26,34-bis(N ε -(7-deoxycholoyl)-GLP-1(7-38), Gly8Lys26(N ε -(7-deoxycholoyl)-GLP-1(7-38), Gly8Lys34(N ε -(7-deoxycholoyl)-GLP-1(7-38), Gly8Lys26,34-bis(N ε -(7-deoxycholoyl)-GLP-1(7-38), Arg26Lys34(N ε-(7-deoxycholoyl)-GLP-1(7-38), Lys26(N ε -(7-deoxycholoyl)-GLP-1(7-39), Lys34(N ε -(7-deoxycholoyl)-GLP-1(7-39), Lys26,34-bis(N ε -(7-deoxycholoyl)-GLP-1(7-39), Gly8Lys26(N ε -(7-deoxycholoyl)-GLP-1(7-39), Gly8Lys34(N ε -(7-deoxycholoyl)-GLP-1(7-39), Gly8Lys26,34-bis(N ε -(7-deoxycholoyl)-GLP-1(7-39), Arg26Lys34(N ε -(7-deoxycholoyl)-GLP-1(7-39), Lys26(N ε -(7-deoxycholoyl))-GLP-1(7-40), Lys34(N ε -(7-deoxycholoyl))-GLP-1(7-40), Lys26,34-bis(N ε -(7-deoxycholoyl))-GLP-1(7-40), Gly8Lys26(N ε -(7-deoxycholoyl))-GLP-1(7-40), Gly8Lys34(N ε -(7-deoxycholoyl))-GLP-1(7-40), Gly8Lys26,34-bis(N ε -(7-deoxycholoyl))-GLP-1(7-40), Arg26Lys34(N ε -(7-deoxycholoyl))-GLP-1(7-40), Lys26(N ε -(7-deoxycholoyl)-GLP-1(7-36), Lys34(N ε -(7-deoxycholoyl)-GLP-1(7-36), Lys26,34-bis(N ε -(7-deoxycholoyl)-GLP-1(7-36), Gly8Lys26(N ε -(7-deoxycholoyl)-GLP-1(7-36), Gly8Lys34(N ε -(7-deoxycholoyl)-GLP-1(7-36), Gly8Lys26,34-bis(N ε -(7-deoxycholoyl)-GLP-1(7-36), Arg26Lys34(N ε -(7-deoxycholoyl)-GLP-1(7-36), Lys26(N ε -(7-deoxycholoyl)-GLP-1(7-35), Lys34(N ε -(7-deoxycholoyl)-GLP-1(7-35), Lys26,34-bis(N ε -(7-deoxycholoyl)-GLP-1(7-35), Gly8Lys26(N ε -(7-deoxycholoyl)-GLP-1(7-35), Gly8Lys34(N ε -(7-deoxycholoyl)-GLP-1(7-35), Gly8Lys26,34-bis(N ε -(7-deoxycholoyl)-GLP-1(7-35), Arg26Lys34(N ε -(7-deoxycholoyl)-GLP-1(7-35), Lys26(N ε -(7-deoxycholoyl)-GLP-1(7-36)amide, Lys34(N ε -(7-deoxycholoyl)-GLP-1(7-36)amide, Lys26,34-bis(N ε-(7-deoxycholoyl)-GLP-1(7-36)amide, Gly8Lys26(N ε -(7-deoxycholoyl)-GLP-1(7-36)amide, Gly8Lys34(N ε -(7-deoxycholoyl)-GLP-1(7-36)amide, Gly8Lys26,34-bis(N ε -(7-deoxycholoyl)-GLP-1(7-36)amide, Arg26Lys34(N ε -(7-deoxycholoyl)-GLP-1(7-36)amide, Gly8Arg26Lys34(N ε -(7-deoxycholoyl)-GLP-1(7-37), Lys26(N ε -(7-deoxycholoyl))Arg34-GLP-1(7-37), Gly8Lys26(N ε -(7-deoxycholoyl))Arg34-GLP-1(7-37), Arg26,34Lys36(N ε -(7-deoxycholoyl)-GLP-1(7-37), Gly8Arg26,34Lys36(N ε -(7-deoxycholoyl)-GLP-1(7-37), Lys26(N ε -(Coloyl)-GLP-1(7-37), Lys34(N ε -(Coloyl)-GLP-1(7-37), Lys26,34-bis(N ε -(Coloyl)-GLP-1(7-37), Gly8Lys26(N ε -(Coloyl)-GLP-1(7-37), Gly8Lys34(N ε -(Coloyl)-GLP-1(7-37), Gly8Lys26,34-bis(N ε -(Coloyl)-GLP-1(7-37), Arg26Lys34(N ε -(Coloyl)-GLP-1(7-37), Gly8Arg26Lys34(N ε -(7-deoxycholoyl)-GLP-1(7-38), Lys26(N ε -(7-deoxycholoyl))Arg34-GLP-1(7-38), Gly8Lys26(N ε -(7-deoxycholoyl))Arg34-GLP-1(7-38), Arg26,34Lys36(N ε -(7-deoxycholoyl)-GLP-1(7-38), Arg26,34Lys36(N ε -(7-deoxycholoyl)-GLP-1(7-38), Gly8Arg26,34Lys36(N ε -(7-deoxycholoyl)-GLP-1(7-38), Lys26(N ε -(Coloyl)-GLP-1(7-38), Lys34(N ε -(Coloyl)-GLP-1(7-38), Lys26,34-bis(N ε -(Coloyl)-GLP-1(7-38), Gly8Lys26(N ε -(Coloyl)-GLP-1(7-38), Gly8Lys34(N ε -(Coloyl)-GLP-1(7-38), Gly8Lys26,34-bis(N ε -(Coloyl)-GLP-1(7-38), Arg26Lys34(N ε -(Coloyl)-GLP-1(7-38), Gly8Arg26Lys34(N ε -(7-deoxycholoyl)-GLP-1(7-39), Lys26(N ε-(7-deoxycholoyl))Arg34-GLP-1(7-39), Gly8Lys26(N ε -(7-deoxycholoyl))Arg34-GLP-1(7-39), Arg26,34Lys36(N ε -(7-deoxycholoyl)-GLP-1(7-39), Gly8Arg26,34Lys36(N ε -(7-deoxycholoyl)-GLP-1(7-39), Lys26(N ε -(Coloyl)-GLP-1(7-39), Lys34(N ε -(Coloyl)-GLP-1(7-39), Lys26,34-bis(N ε -(Coloyl)-GLP-1(7-39), Gly8Lys26(N ε -(Coloyl)-GLP-1(7-39), Gly8Lys34(N ε -(Coloyl)-GLP-1(7-39), Gly8Lys26,34-bis(N ε -(Coloyl)-GLP-1(7-39), Arg26Lys34(N ε -(Coloyl)-GLP-1(7-39), Gly8Arg26Lys34(N ε -(7-deoxycholoyl))-GLP-1(7-40), Lys26(N ε -(7-deoxycholoyl))Arg34-GLP-1(7-40), Gly8Lys26(N ε -(7-deoxycholoyl))Arg34-GLP-1(7-40), Arg26,34Lys36(N ε -(7-deoxycholoyl))-GLP-1(7-40), Gly8Arg26,34Lys38(N ε -(7-deoxycholoyl))-GLP-1(7-40), Lys26(N ε -(Coloil))-G LP-1(740), Lys34(N ε -(Coloil))-GLP-1(740), Lys26,34-bis(N ε -(Coloyl)-GLP-1(7-40), Gly8Lys26(N ε -(Coloyl)-GLP-1(7-40), Gly8Lys34(N ε -(Coloyl)-GLP-1(7-40), Gly8Lys26,34-bis(N ε -(Coloyl)-GLP-1(7-40), Arg26Lys34(N ε -(Coloyl)-GLP-1(7-40), Lys26(N ε -(Coloyl)-GLP-1(7-36), Lys34(N ε -(Coloyl)-GLP-1(7-36), Lys26,34-bis(N ε -(Coloyl)-GLP-1(7-36), Gly8Lys26(N ε -(Coloyl)-GLP-1(7-36), Gly8Lys34(N ε -(Coloyl)-GLP-1(7-36), Gly8Lys26,34-bis(N ε -(Coloyl)-GLP-1(7-36), Arg26Lys34(N ε -(Coloyl)-GLP-1(7-36), Lys26(N ε -(Coloyl)-GLP-1(7-35), Lys34(N ε -(Coloyl)-GLP-1(7-35), Lys26,34-bis(N ε -(Coloyl)-GLP-1(7-35), Gly8Lys26(N ε -(Coloyl)-GLP-1(7-35), Gly8Lys34(N ε -(Coloyl)-GLP-1(7-35), Gly8Lys26,34-bis(N ε -(Coloyl)-GLP-1(7-35), Arg26Lys34(N ε -(Coloyl)-GLP-1(7-35), Lys26(N ε -(choloyl)-GLP-1(7-36)amide, Lys34(N ε -(choloyl)-GLP-1(7-36)amide, Lys26,34-bis(N ε -(choloyl)-GLP-1(7-36)amide, Gly8Lys26(N-(choloyl))-GLP-1(7-36)amide, Gly8Lys34(N ε -(choloyl)-GLP-1(7-36)amide, Gly8Lys26,34-bis(N ε -(choloyl)-GLP-1(7-36)amide, Arg26Lys34(N ε -(choloyl)-GLP-1(7-36)amide, Gly8Arg26Lys34(N ε -(Coloyl)-GLP-1(7-37), Lys26(N ε -(Coloyl))Arg34-GLP-1(7-37), Gly8Lys26(N ε -(Coloyl))ArS34-GLP-1(7-37), Arg26,34Lys36(N ε -(Coloyl)-GLP-1(7-37), Gly8Arg26,34Lys36(N ε -(Coloyl)-GLP-1(7-37), Lys26(N ε-(lithocoloyl)-GLP-1(7-37), Lys34(N ε -(lithocoloyl)-GLP-1(7-37), Lys26,34-bis(N ε -(lithocoloyl)-GLP-1(7-37), Gly8Lys26(N ε -lithocoloyl))-GLP-1(7-37), Gly8Lys34(N ε -(lithocoloyl)-GLP-1(7-37), Gly8Lys26,34-bis(N ε -(lithocoloyl)-GLP-1(7-37), Arg26Lys34(N ε -(lithocoloyl)-GLP-1(7-37), Gly8Arg26Lys34(N ε -(Coloyl)-GLP-1(7-38), Lys26(N ε -(Coloyl))Arg34-GLP-1(7-38), Gly8Lys26(N ε -(Coloyl))Arg34-GLP-1(7-38), Arg26,34Lys36(N ε -(Coloyl)-GLP-1(7-38), Arg26,34Lys38(N ε -(Coloyl)-GLP-1(7-38), Gly8Arg26,34Lys36(N ε -(Coloyl)-GLP-1(7-38), Lys26(N ε -(lithocoloyl)-GLP-1(7-38), Lys34(N ε -(lithocoloyl)-GLP-1(7-38), Lys26,34-bis(N ε -(lithocoloyl)-GLP-1(7-38), Gly8Lys26(N ε -(lithocoloyl)-GLP-1(7-38), Gly8Lys34(N ε -(lithocoloyl)-GLP-1(7-38), Gly8Lys26,34-bis(N ε -(lithocoloyl)-GLP-1(7-38), Arg26Lys34(N ε -(lithocoloyl)-GLP-1(7-38), Gly8Arg26Lys34(N ε -(Coloyl)-GLP-1(7-39), Lys26(N ε -(Coloyl))Arg34-GLP-1(7-39), Gly8Lys26(N ε -(Coloyl))Arg34-GLP-1(7-39), Arg26,34Lys36(N ε -(Coloyl)-GLP-1(7-39), Gly8Arg26.34Lys36(N ε -(Coloyl)-GLP-1(7-39), Lys26(N ε -(lithocoloyl)-GLP-1(7-39), Lys34(N ε -(lithocoloyl)-GLP-1(7-39), Lys26,34-bis(N ε -(lithocoloyl)-GLP-1(7-39), Gly8Lys26(N ε -(lithocoloyl)-GLP-1(7-39), Gly8Lys34(N ε -(lithocoloyl)-GLP-1(7-39), Gly8Lys26,34-bis(N ε -(lithocoloyl)-GLP-1(7-39), Arg26Lys34(N ε -(lithocoloyl)-GLP-1(7-39), Gly8Arg26Lys34(N ε -(Coloyl)-GLP-1(7-40), Lys26(N ε -(Coloyl))Arg34-GLP-1(7-40), Gly8Lys26(N ε -(Coloyl))Arg34-GLP-1(7-40), Arg26,34Lys36(N ε -(Coloyl)-GLP-1(7-40), Arg26,34Lys38(N ε -(Coloyl)-GLP-1(7-40), Gly8Arg26,34Lys36(N ε -(Coloyl)-GLP-1(7-40), Lys26(N ε -(lithocoloyl)-GLP-1(7-40), Lys34(N ε -(lithocoloyl)-GLP-1(7-40), Lys26,34-bis(N ε -(lithocoloyl)-GLP-1(7-40), Gly8Lys26(N ε -(lithocoloyl)-GLP-1(7-40), Gly8Lys34(N ε -(lithocoloyl)-GLP-1(7-40), Gly8Lys26,34-bis(N ε -(lithocoloyl)-GLP-1(7-40), Arg26Lys34(N ε -(lithocoloyl)-GLP-1(7-37), Lys26(N ε -(lithocoloyl)-GLP-1(7-36), Lys34(N ε -(lithocoloyl)-GLP-1(7-36), Lys26,34-bis(N ε -(lithocoloyl)-GLP-1(7-36), Gly8Lys26(N ε -lithocoloyl))-GLP-1(7-36), Gly8Lys34(N ε-(lithocoloyl)-GLP-1(7-36), Gly8Lys26,34-bis(N ε -(lithocoloyl)-GLP-1-(7-36), Arg26Lys34(N ε -(lithocoloyl)-GLP-1(7-36), Lys26(N ε -(lithocoloyl)-GLP-1(7-35), Lys34(N ε -(lithocoloyl)-GLP-1(7-35), Lys26,34-bis(N ε -(lithocoloyl)-GLP-1(7-35), Gly8Lys26(N ε -(lithocoloyl)-GLP-1(7-35), Gly8Lys34(N ε -(lithocoloyl)-GLP-1(7-35), Gly8Lys26,34-bis(N ε -(lithocoloyl)-GLP-1(7-35), Arg26Lys34(N ε -(lithocoloyl)-GLP-1(7-35), Lys26(N ε -(lithocoloyl)-GLP-1-(7-36)amide, Lys34(N ε -(lithocoloyl)-GLP-1(7-36)amide, Lys26,34-bis(N ε -(lithocoloyl)-GLP-1(7-36)amide, Gly8Lys26(N ε -(lithocoloyl)-GLP-1(7-36)amide, Gly8Lys34(N ε -(lithocoloyl)-GLP-1(7-36)amide, Gly8Lys26,34-bis(N ε -(lithocoloyl)-GLP-1(7-36)amide, Arg26Lys34(N ε-(lithocoloyl)-GLP-1(7-36)amide, Gly8Arg26Lys34(N ε -(lithocoloyl)-GLP-1(7-37), Lys26(N ε -(lithocoloyl))Arg34-GLP-1(7-37), Gly8Lys26(N ε -(lithocoloyl))Arg34-GLP-1(7-37), Arg26,34Lys36(N ε -(lithocoloyl)-GLP-1(7-37), Gly8Arg26,34Lys36(N ε -(lithocoloyl)-GLP-1(7-37), Gly8Arg26Lys34(N ε -(lithocoloyl)-GLP-1(7-38), Lys26(N ε -(lithocoloyl))AFS34-GLP-1(7-38), Gly8Lys26(N ε -(lithocoloyl))Arg34-GLP-1(7-38), Arg26,34Lys36(N ε -(lithocoloyl)-GLP-1(7-38), Arg26,34Lys36(N ε -(lithocoloyl)-GLP-1(7-38), Gly8Arg26,34Lys36(N ε -(lithocoloyl)-GLP-1(7-38), Gly8Arg26Lys34(N ε -(lithocoloyl)-GLP-1(7-39), Lys26(N ε -(lithocoloyl))Arg34-GLP-1(7-39), Gly8Lys26(N ε -(lithocoloyl))Arg34-GLP-1(7-39), Arg26,34Lys36(N ε -(lithocoloyl)-GLP-1(7-39), Gly8Arg26,34Lys36(N ε -(lithocoloyl)-GLP-1(7-39), Gly8Arg26Lys34(N ε -(lithocoloyl)-GLP-1(7-40), Lys26(N ε -(lithocoloyl))Arg34-GLP-1(7-40), Gly8Lys26(N ε -(lithocoloyl))Arg34-GLP-1(7-40), Arg26,34Lys36(N ε -(lithocoloyl)-GLP-1(7-40) and Gly8Arg26,34Lys36(N ε -(lithocoloyl)-GLP-1(7-40), The GLP-1 derivative may be selected from the group consisting of:

[0247] In certain embodiments, the therapeutic polypeptide may be a GLP-1 agonist disclosed in WO 2012 / 080471 or WO 2013 / 189988, such as semaglutide and structurally related analogues thereof. Semaglutide has the structure shown in Formula II and Figure 16B.

[0248] [ka]

[0249] In some embodiments, a simplified nomenclature is used to describe GLP-1 agonists, e.g., [Aib8]GLP-1(7-37) refers to an analog of GLP-1(7-37) in which the native Ala at position 8 is replaced with Aib. In some embodiments, the GLP-1 agonist comprises up to 12 amino acids, e.g., up to 10, 8, or 6, that have been altered, e.g., by substitution, deletion, insertion, and / or modification, e.g., compared to GLP-1(7-37). In some embodiments, the analog comprises up to 10 substitutions, deletions, additions, and / or insertions, e.g., up to 9 substitutions, deletions, additions, and / or insertions, up to 8 substitutions, deletions, additions, and / or insertions, up to 7 substitutions, deletions, additions, and / or insertions, up to 6 substitutions, deletions, additions, and / or insertions, up to 5 substitutions, deletions, additions, and / or insertions, up to 4 substitutions, deletions, additions, and / or insertions, or up to 3 substitutions, deletions, additions, and / or insertions, e.g., compared to GLP-1(7-37). Unless otherwise specified, GLP-1 includes only L-amino acids.

[0250] In some embodiments, the terms "GLP-1 agonist," "GLP-1 receptor agonist," "GLP-1 analog," or "GLP-1 analog," as used interchangeably herein, refer to a peptide or compound that is a variant of human glucagon-like peptide-1 (GLP-1(7-37)). GLP-1(7-37) has the sequence HAEGTFTSDVSSYLEGQAAKEFIAWLVKGRG. In some embodiments, the term "variant" refers to a compound that contains one or more amino acid substitutions, deletions, additions, and / or insertions.

[0251] In one embodiment, the GLP-1 agonist exhibits at least 60%, 65%, 70%, 80%, or 90% sequence identity with GLP-1(7-37) over the entire length of GLP-1(7-37). As an example of a method for determining sequence identity between two analogs, two peptides, [Aib8]GLP-1(7-37) and GLP-1(7-37), are aligned. The sequence identity of [Aib8]GLP-1(7-37) compared to GLP-1(7-37) is given by the number of aligned identical residues minus the number of different residues, divided by the total number of residues in GLP-1(7-37). Thus, in the above example, the sequence identity is (31-1) / 31.

[0252] In one embodiment, the C-terminus of the GLP-1 agonist is an amide.

[0253] In some embodiments, the GLP-1 agonist is GLP-1(7-37) or GLP-1(7-36)amide. In some embodiments, the GLP-1 agonist is exendin-4, the sequence of which is HGEGTFTSDLSKQMEEEAVRLFIEWLKNGGPSSGAPPPS.

[0254] In some embodiments, the GLP-1 agonist comprises one substituent covalently attached to the peptide. In some embodiments, the substituent comprises a fatty acid or an aliphatic diacid. In some embodiments, the substituent comprises a C16, C18, or C20 fatty acid. In some embodiments, the substituent comprises a C16, C18, or C20 aliphatic diacid. In some embodiments, the substituent is

[0255] [ka]

[0256] In some embodiments, the substituent is represented by formula (X) and n is at least 13, e.g., n is 13, 14, 15, 16, 17, 18, or 19. In some embodiments, the substituent is represented by formula (X) and n is in the range of 13 to 19, e.g., in the range of 13 to 17. In some embodiments, the substituent is represented by formula (X) and n is 13, 15, or 17. In some embodiments, the substituent is represented by formula (X) and n is 13. In some embodiments, the substituent is represented by formula (X) and n is 15. In some embodiments, the substituent is represented by formula (X) and n is 17. In some embodiments, the substituent comprises one or more 8-amino-3,6-dioxaoctanoic acid (OEG), e.g., two OEGs.

[0257] In some embodiments, the substituent is [2-(2-{2-[2-(2-{2-[(S)-4-carboxy-4-(17-carboxyheptadecanoylamino)butyrylamino]ethoxy}ethoxy)acetylamino]ethoxy}ethoxy)acetyl].

[0258] In some embodiments, the substituent is [2-(2-{2-[2-(2-{2-[(S)-4-carboxy-4-({trans-4-[(19-carboxynonadecanoylamino)methyl]cyclohexanecarbonyl}amino)butyrylamino]ethoxy}ethoxy)acetylamino]ethoxy}ethoxy)acetyl].

[0259] In some embodiments, the GLP-1 agonist is semaglutide, also known as N-ε26-[2-(2-{2-[2-(2-{2-[(S)-4-carboxy-4-(17-carboxyheptadecanoylamino)butyrylamino]ethoxy}ethoxy)acetylamino]ethoxy}ethoxy)acetyl][Aib8, Arg34]GLP-1(7-37), which may be prepared as described in Example 4 of WO 2006 / 097537, which is incorporated herein by reference.

[0260] In some embodiments, the GLP-1 agonist is in the form of a pharmaceutically acceptable salt, amide, or ester thereof. In some embodiments, the GLP-1 agonist includes one or more pharmaceutically acceptable counterions.

[0261] In some embodiments, the dosage of the GLP-1 agonist is in the range of 0.01 mg to 100 mg. In some embodiments, the composition for administering a GLP-1 agonist comprises an amount of the GLP-1 agonist in the range of 0.1 to 40 mg or 1 to 20 mg. In some embodiments, the composition comprises an amount of the GLP-1 agonist in the range of 5 to 20 mg, such as in the range of 5 to 15 mg, such as 5 mg, for example 10 mg, for example 15 mg, for example 20 mg.

[0262] In some embodiments, the composition comprises an amount of the GLP-1 agonist in the range of 0.05 to 25 μmol, such as in the range of 0.5 to 2.5 pmol.

[0263] In some embodiments, the GLP-1 agonist is a GLP-1 agonist as described in WO 93 / 19175, WO 96 / 29342, WO 98 / 08871, WO 99 / 43707, WO 99 / 43706, WO 99 / 43341, WO 99 / 43708, WO 2005 / 027978, WO 2005 / 058954, WO 2005 / 058958, WO 2006 / 005667, WO 2006 / 037810, WO 2006 / 037811, WO 2006 / 097 537, WO2006 / 097538, WO2008 / 023050, WO2009 / 030738, WO2009 / 030771, WO2009 / 030774, WO2011 / 080102, WO2011 / 080103, WO2012 / 062803, WO2012 / 062804, WO2012 / 140117, WO2014 / 005858, WO2014 / 177683 and WO2015 / 000942.

[0264] In some embodiments, the GLP-1 agonist is, for example,

[0265] N-ε37{2-[2-(2-{2-[2-((R)-3-carboxy-3-{[1-(19-carboxynonadecanoyl)piperidine-4-carbonyl]amino}propionylamino)ethoxy]ethoxy}acetylamino)ethoxy]ethoxy}acetyl[desaminoHis7, Glu22, Arg26, Arg34, Lys37]GLP-1(7-37)amide,

[0266] N-ε26{2-[2-(2-{2-[2-((R)-3-carboxy-3-{[1-(19-carboxynonadecanoyl)piperidine-4-carbonyl]amino}propionylamino)ethoxy]ethoxy}acetylamino)ethoxy]ethoxy}acetyl[desaminoHis7,Arg34]GLP-1-(7-37),

[0267] N-ε37{2-[2-(2-{2-[2-((S)-3-carboxy-3-{[1-(19-carboxynonadecanoyl)piperidine-4-carbonyl]amino}propionylamino)ethoxy]ethoxy}acetylamino)ethoxy]ethoxy}acetyl[Aib8, Glu22, Arg26, Arg34, Lys37]GLP-1-(7-37)amide,

[0268] N-ε37-[2-(2-[2-(2-[2-(2-((R)-3-[1-(17-carboxyheptadecanoyl)piperidin-4-ylcarbonylamino]3-carboxypropionylamino)ethoxy)ethoxy]acetylamino)ethoxy]ethoxy)acetyl][desaminoHis7, Glu22, Arg26, Arg 34, Phe(m-CF3)28]GLP-1-(7-37)amide,

[0269] N-ε26-[(S)-4-carboxy-4-({trans-4-[(19-carboxynonadecanoylamino)methyl]cyclohexanecarbonyl}amino)butyryl][Aib8, Arg34]GLP-1-(7-37),

[0270] N-ε26-{4-[(S)-4-carboxy-4-({trans-4-[(19-carboxynonadecanoylamino)methyl]cyclohexanecarbonyl}amino)butyrylamino]butyryl}[Aib8, Arg34]GLP-1-(7-37),

[0271] N-ε26-[2-(2-{2-[(S)-4-carboxy-4-({trans-4-[(19-carboxynonadecanoylamino)methyl]cyclohexanecarbonyl}amino)butyrylamino]ethoxy}ethoxy)acetyl][Aib8, Arg34]GLP-1-(7-37),

[0272] N-ε26-[2-(2-{2-[2-(2-{2-[(S)-4-carboxy-4-({trans-4-[(19-carboxynonadecanoylamino)methyl]cyclohexanecarbonyl}amino)butyrylamino]ethoxy}ethoxy)acetylamino]ethoxy}ethoxy)acetyl][Aib8, Arg34]GLP-1-(7-37)amide,

[0273] N-ε37-[2-(2-{2-[2-(2-{2-[(S)-4-carboxy-4-({trans-4-[(19-carboxynonadecanoylamino)methyl]cyclohexanecarbonyl}amino)butyrylamino]ethoxy}ethoxy)acetylamino]ethoxy}ethoxy)acetyl][Aib8, Glu22, Arg26, Arg34, Lys37)]GLP-1-(7-37)amide,

[0274] N-ε37-[2-(2-{2-[2-(2-{2-[(S)-4-carboxy-4-({trans-4-[(19-carboxynonadecanoylamino)methyl]cyclohexanecarbonyl}amino)butyrylamino]ethoxy}ethoxy)acetylamino]ethoxy}ethoxy)acetyl][desaminoHis7, Glu22, Arg26, Arg34, Lys37]GLP-1-(7-37)amide,

[0275] N-ε37-[2-(2-{2-[2-(2-{2-[(S)-4-carboxy-4-({4-[(trans-19-carboxynonadecanoylamino)methyl]cyclohexanecarbonyl}amino)butyrylamino]ethoxy}ethoxy)acetylamino]ethoxy}ethoxy)acetyl][desaminoHis7, Arg26, Arg34, Lys37]GLP-1-(7-37)amide,

[0276] N-ε37-[2-(2-{2-[2-(2-{2-[(S)-4-carboxy-4-({trans-4-[(19-carboxynonadecanoylamino)methyl]cyclohexanecarbonyl}amino)butyrylamino]ethoxy}ethoxy)acetylamino]ethoxy}ethoxy)acetyl][desaminoHis7, Glu22, Arg26, Arg34, Lys37]GLP-1-(7-37),

[0277] N-ε26[2-(2-{2-[2-(2-{2-[(S)-4-carboxy-4-({4-[(19-carboxynonadecanoylamino)methyl]cyclohexanecarbonyl}amino)butyrylamino]ethoxy}ethoxy)acetylamino]ethoxy}ethoxy)acetyl[Aib8, Lys26]GLP-1(7-37)amide,

[0278] N-ε26[2-(2-[2-(2-[2-(2-(2-((S)-2-[trans-4-((9-carboxynonadecanoylamino]methyl)cyclohexylcarbonylamino]-4-carboxybutanoylamino)ethoxy)ethoxy]acetylamino)ethoxy]ethoxy)acetyl]GLP-1[Aib8, Lys26](7-37)amide,

[0279] N-ε37-[2-(2-{2-[2-(2-{2-[(S)-4-carboxy-4-({trans-4-[(19-carboxynonadecanoylamino)methyl]cyclohexanecarbonyl}amino)butyrylamino]ethoxy}ethoxy)acetylamino]ethoxy}ethoxy)acetyl][desaminoHis7, Arg26, Arg34, Lys37]GLP-1-(7-37),

[0280] N-ε37-[2-(2-{2-[2-(2-{2-[(S)-4-carboxy-4-({trans-4-[(19-carboxynonadecanoylamino)methyl]cyclohexanecarbonyl}amino)butyrylamino]ethoxy}ethoxy)acetylamino]ethoxy}ethoxy)acetyl][desaminoHis7, Glu22, Arg26, Glu30, Arg34, Lys37]GLP-1-(7-37),

[0281] N-ε26-[2-(2-{2-[(S)-4-carboxy-4-((S)-4-carboxy-4-{4-[4-(16-(1H-tetrazol-5-yl)hexadecanoylsulfamoyl)butyrylamino]butyrylamino}butyrylamino)butyrylamino]ethoxy}ethoxy)acetyl][Aib8, Arg34]GLP-1-(7-37),

[0282] N-ε26-[2-(2-{2-[(S)-4-carboxy-4-((S)-4-carboxy-4-{12-[4-(16-(1H-tetrazol-5-yl)hexadecanoylsulfamoyl)butyrylamino]dodecanoylamino}butyrylamino)butyrylamino]ethoxy}ethoxy)acetyl][Aib8, Arg34]GLP-1-(7-37),

[0283] N-ε26-[2-(2-{2-[(S)-4-carboxy-4-((S)-4-carboxy-4-{6-[4-(16-(1H-tetrazol-5-yl)hexadecanoylsulfamoyl)butyrylamino]hexanoylamino}butyrylamino)butyrylamino]ethoxy}ethoxy)acetyl][Aib8, Arg34]GLP-1-(7-37),

[0284] N-ε26-[2-(2-{2-[(S)-4-carboxy-4-((S)-4-carboxy-4-{4-[4-(16-(1H-tetrazol-5-yl)hexadecanoylsulfamoyl)butyrylamino]butyrylamino}butyrylamino)butyrylamino]ethoxy}ethoxy)acetyl][Aib8, Arg34]GLP-1-(7-34),

[0285] N-ε26-[2-(2-{2-[(S)-4-carboxy-4-((S)-4-carboxy-4-{12-[4-(16-(1H-tetrazol-5-yl)hexadecanoylsulfamoyl)butyrylamino]dodecanoylamino}butyrylamino)butyrylamino]ethoxy}ethoxy)acetyl][Aib8, Arg34]GLP-1-(7-34),

[0286] N-ε26-[2-(2-{2-[(S)-4-carboxy-4-((S)-4-carboxy-4-{6-[4-(16-(1H-tetrazol-5-yl)hexadecanoylsulfamoyl)butyrylamino]hexanoylamino}butyrylamino)butyrylamino]ethoxy}ethoxy)acetyl][Aib8, Arg34]GLP-1-(7-34),

[0287] N-ε26-[2-(2-{2-[(S)-4-carboxy-4-((S)-4-carboxy-4-{12-[4-(16-(1H-tetrazol-5-yl)hexadecanoylsulfamoyl)butyrylamino]dodecanoylamino}butyrylamino)butyrylamino]ethoxy}ethoxy)acetyl][Aib8, Arg34]GLP-1-(7-35),

[0288] N-ε26-[2-(2-{2-[(S)-4-carboxy-4-((S)-4-carboxy-4-{6-[4-(16-(1H-tetrazol-5-yl)hexadecanoylsulfamoyl)butyrylamino]hexanoylamino}butyrylamino)butyrylamino]ethoxy}ethoxy)acetyl][Aib8, Arg34]GLP-1-(7-35),

[0289] N-ε26-[2-(2-{2-[(S)-4-carboxy-4-((S)-4-carboxy-4-{6-[4-(16-(1H-tetrazol-5-yl)hexadecanoylsulfamoyl)butyrylamino]hexanoylamino}butyrylamino)butyrylamino]ethoxy}ethoxy)acetyl][Aib8, Arg34]GLP-1-(7-36)amide,

[0290] N-ε26-[2-(2-{2-[(S)-4-carboxy-4-((S)-4-carboxy-4-{6-[4-(16-(1H-tetrazol-5-yl)hexadecanoylsulfamoyl)butyrylamino]hexanoylamino}butyrylamino)butyrylamino]ethoxy}ethoxy)acetyl][Aib8, Arg34]GLP-1-(7-35),

[0291] N-ε26-[2-(2-{2-[(S)-4-carboxy-4-((S)-4-carboxy-4-{12-[4-(16-(1H-tetrazol-5-yl)hexadecanoylsulfamoyl)butyrylamino]dodecanoylamino}butyrylamino)butyrylamino]ethoxy}ethoxy)acetyl][Aib8, Lys33, Arg34]GLP-1-(7-34),

[0292] N-ε26-[2-(2-{2-[(S)-4-carboxy-4-((S)-4-carboxy-4-{12-[4-(16-(1H-tetrazol-5-yl)hexadecanoylsulfamoyl)butyrylamino]dodecanoylamino}butyrylamino)butyrylamino]ethoxy}ethoxy)acetyl][Aib8, Arg34]GLP-1-(7-36)amide,

[0293] N-ε26-[2-(2-{2-[2-(2-{2-[2-(2-{2-[2-(2-{2-[2-(2-{2-[2-(2-{2-[2-(2-{2-[2-(2-{2-[(S)-4-carboxy-4-((S)-4-carboxy-4-{12-[4-(16-(1H-tetrazol-5-yl)hexadecanoylsulfamoyl)butyrylamino]dodecanoylamino}butyrylamino)butyrylamino]ethoxy}ethoxy)acetylamino]ethoxy}ethoxy)acetylamino]ethoxy}ethoxy)acetylamino]ethoxy}ethoxy)acetylamino]ethoxy}ethoxy)acetylamino]ethoxy}ethoxy)acetylamino]ethoxy}ethoxy)acetyl][Aib8, Lys26, Arg34]GLP-1-(7-36)amide,

[0294] N-ε37-[2-(2-{2-[(S)-4-carboxy-4-((S)-4-carboxy-4-{12-[4-(16-(1H-tetrazol-5-yl)hexadecanoylsulfamoyl)butyrylamino]dodecanoylamino}butyrylamino)butyrylamino]ethoxy}ethoxy)acetyl][Aib8, Glu22, Arg26, Arg34, Lys37]GLP-1-(7-37)amide,

[0295] N-ε37-[2-(2-{2-[(S)-4-carboxy-4-((S)-4-carboxy-4-{12-[4-(16-(1H-tetrazol-5-yl)hexadecanoylsulfamoyl)butyrylamino]dodecanoylamino}butyrylamino)butyrylamino]ethoxy}ethoxy)acetyl][desaminoHis7, Glu22, Arg26, Arg34, Lys37]GLP-1-(7-37)amide,

[0296] N-ε37{2-[2-(2-{2-[2-((R)-3-carboxy-3-{[1-(19-carboxynonadecanoyl)piperidine-4-carbonyl]amino}propionylamino)ethoxy]ethoxy}acetylamino)ethoxy]ethoxy}acetyl[desaminoHis7, Glu22, Arg26, Arg34, Lys37]GLP-1(7-37)amide,

[0297] N-ε37{2-[2-(2-{2-[2-((S)-3-carboxy-3-{[1-(19-carboxynonadecanoyl)piperidine-4-carbonyl]amino}propionylamino)ethoxy]ethoxy}acetylamino)ethoxy]ethoxy}acetyl[Aib8, Glu22, Arg26, Arg34, Lys37]GLP-1-(7-37)amide,

[0298] N-ε37-[2-(2-[2-(2-[2-(2-((R)-3-[1-(17-carboxyheptadecanoyl)piperidin-4-ylcarbonylamino]3-carboxypropionylamino)ethoxy)ethoxy]acetylamino)ethoxy]ethoxy)acetyl][desaminoHis7, Glu22, Arg26, Arg34, Phe(m-CF3)28]GLP-1-(7-37)amide,

[0299] N-ε37-[2-(2-{2-[2-(2-{2-[(S)-4-carboxy-4-({trans-4-[(19-carboxy-nonadecanoylamino)methyl]cyclohexanecarbonyl}amino)butyrylamino]ethoxy}ethoxy)acetylamino]ethoxy}ethoxy)acetyl][Aib8, Glu22, Arg26, Arg34, Lys37]GLP-1-(7-37)amide,

[0300] N-ε37-[2-(2-{2-[2-(2-{2-[(S)-4-carboxy-4-({trans-4-[(19-carboxynonadecanoylamino)methyl]cyclohexanecarbonyl}amino)butyrylamino]ethoxy}ethoxy)acetylamino]ethoxy}ethoxy)acetyl][desaminoHis7, Glu22, Arg26, Arg34, Lys37]GLP-1-(7-37)amide,

[0301] N-ε37-[2-(2-{2-[2-(2-{2-[(S)-4-carboxy-4-({trans-4-[(19-carboxynonadecanoylamino)methyl]cyclohexanecarbonyl}amino)butyrylamino]ethoxy}ethoxy)acetylamino]ethoxy}ethoxy)acetyl][desaminoHis7, Glu22, Arg26, Arg34, Lys37]GLP-1-(7-37),

[0302] N-ε37-[2-(2-{2-[2-(2-{2-[(S)-4-carboxy-4-({trans-4-[(19-carboxynonadecanoylamino)methyl]cyclohexanecarbonyl}amino)butyrylamino]ethoxy}ethoxy)acetylamino]ethoxy}ethoxy)acetyl][desaminoHis7, Glu22, Arg26, Glu30, Arg34, Lys37]GLP-1-(7-37),

[0303] N-ε37-[2-(2-{2-[(S)-4-carboxy-4-((S)-4-carboxy-4-{12-[4-(16-(1H-tetrazol-5-yl)hexadecanoylsulfamoyl)butyrylamino]dodecanoylamino}butyrylamino)butyrylamino]ethoxy}ethoxy)acetyl][Aib8, Glu22, Arg26, Arg34, Lys37]GLP-1-(7-37)amide,

[0304] N-ε37-[2-(2-{2-[(S)-4-carboxy-4-((S)-4-carboxy-4-{12-[4-(16-(lH-tetrazol-5-yl)hexadecanoylsulfamoyl)butyrylamino]dodecanoylamino}butyrylamino)butyrylamino]ethoxy}ethoxy)acetyl][desaminoHis7, Glu22, Arg26, Arg34, Lys37]GLP-1-(7-37)amide,

[0305] N-ε37-(3-((2-(2-(2-(2-(2-hexadecyloxyethoxy)ethoxy)ethoxy)ethoxy)ethoxy)propionyl)[desaminoHis7, Glu22, Arg26, Arg34, Lys37]GLP-1(7-37)-amide,

[0306] N-ε37-{2-(2-(2-(2-[2-(2-(4-hexadecanoylamino-4-carboxybutyrylamino)ethoxy)ethoxy]acetyl)ethoxy)ethoxy)acetyl)}[desaminoHis7, Glu22, Arg26, Glu30, Arg34, Lys37]GLP-1-(7-37)amide,

[0307] N-ε37-{2-(2-(2-(2-[2-(2-(4-hexadecanoylamino-4-carboxybutyrylamino)ethoxy)ethoxy]acetyl)ethoxy)ethoxy)acetyl)}[desaminoHis7, Glu22(Arg26, Arg34, Lys37)]GLP-1-(7-37)amide,

[0308] N-ε37-(2-(2-(2-(2-(2-(2-(2-(2-(2-(2-(2-(2-octadecanoylamino)ethoxy)ethoxy)acetylamino)ethoxy)ethoxy)acetylamino)ethoxy)ethoxy)acetyl)[desaminoHis7, Glu22, Arg26, Arg34, Lys37]GLP-1(7-37)amide,

[0309] N-ε37-[4-(16-(1H-tetrazol-5-yl)hexadecanoylsulfamoyl)butyryl][desamino-His7, Glu22, Arg26, Arg34, Lys37]GLP-1-(7-37)amide,

[0310] N-ε37-[2-(2-{2-[2-(2-{2-[(S)-4-carboxy-4-(19-carboxynonadecanoylamino)butyrylamino]ethoxy}ethoxy)acetylamino]ethoxy}ethoxy)acetyl][desaminoHis7, Glu22, Arg26, Arg34, Lys37]GLP-1-(7-37),

[0311] N-ε37-(2-{2-[2-((S)-4-carboxy-4-{(S)-4-carboxy-4-[(S)-4-carboxy-4-(19-carboxynonadecanoylamino)butyrylamino]butyrylamino}butyrylamino)ethoxy]ethoxy}acetyl)[desaminoHis7, Glu22, Arg26, Arg34, Lys37]GLP-1-(7-37),

[0312] N-ε37-{2-[2-(2-{(S)-4-[(S)-4-(12-{4-[16-(2-tert-butyl-2H-tetrazol-5-yl)hexadecanoylsulfamoyl]butyrylamino}dodecanoylamino)-4-carboxybutyrylamino]-4-carboxybutyrylamino}ethoxy)ethoxy]acetyl}[desaminoHis7, Glu22, Arg26, Arg34, Lys37]GLP-1(7-37),

[0313] N-ε37-[2-(2-{2-[2-(2-{2-[(S)-4-carboxy-4-(17-carboxyheptadecanoylamino)butyrylamino]ethoxy}ethoxy)acetylamino]ethoxy}ethoxy)acetyl][Aib8, Glu22, Arg26, Arg34, Lys37]GLP-1-(7-37),

[0314] N-α37-[2-(2-{2-[2-(2-{2-[(S)-4-carboxy-4-(17-carboxyheptadecanoylamino)butyrylamino]ethoxy}ethoxy)acetylamino]ethoxy}ethoxy)acetyl][Aib8, Glu22, Arg26, Arg34, ε-Lys37]GLP-1-(7-37) peptide,

[0315] N-ε37-[2-(2-{2-[2-(2-{2-[(S)-4-carboxy-4-(17-carboxyheptadecanoylamino)butyrylamino]ethoxy}ethoxy)acetylamino]ethoxy}ethoxy)acetyl][desaminoHis7, Glu22, Arg26, Arg34, Lys37]GLP-1-(7-37),

[0316] N-ε36-[2-(2-{2-[2-(2-{2-[(S)-4-carboxy-4-(15-carboxypentadecanoylamino)butyrylamino]ethoxy}ethoxy)acetylamino]ethoxy}ethoxy)acetyl][desaminoHis7, Glu22, Arg26, Glu30, Arg34, Lys36]GLP-1-(7-37)-Glu-Lys peptide,

[0317] N-ε37-[2-(2-{2-[2-(2-{2-[(S)-4-carboxy-4-({trans-4-[(19-carboxynonadecanoylamino)methyl]cyclohexanecarbonyl}amino)butyrylamino]ethoxy}ethoxy)acetylamino]ethoxy}ethoxy)acetyl][Aib8, Glu22, Arg26, Arg34, Lys37]GLP-1-(7-37),

[0318] N-ε37-[2-(2-{2-[2-(2-{2-[(S)-4-carboxy-4-(17-carboxyheptadecanoylamino)butyrylamino]ethoxy}ethoxy)acetylamino]ethoxy}ethoxy)acetyl][Aib8, Glu22, Arg26, Arg34, Aib35, Lys37]GLP-1-(7-37),

[0319] N-ε37-[(S)-4-carboxy-4-(2-{2-[2-(2-{2-[2-(17-carboxyheptadecanoylamino)ethoxy]ethoxy}acetylamino)ethoxy]ethoxy}acetylamino)butyryl][Aib8, Glu22, Arg26, 34, Lys37]GLP-1(7-37),

[0320] N-ε37-[2-(2-[2-(2-[2-(2-[4-(17-carboxyheptadecanoylamino)-4(S)-carboxybutyrylamino]ethoxy)ethoxy]acetylamino)ethoxy]ethoxy)acetyl][ImPr7, Glu22, Arg26, 34, Lys37]GLP-1-(7-37),

[0321] N-ε37-{2-[2-(2-{2-[2-(2-{(S)-4-carboxy-4-[10-(4-carboxyphenoxy)decanoylamino]butyrylamino}ethoxy)ethoxy]acetylamino}ethoxy)ethoxy]acetyl}[Aib8, Arg34, Lys37]GLP-1(7-37)-OH,

[0322] N-ε26(17-carboxyheptadecanoyl)[Aib8,Arg34]GLP-1-(7-37) peptide,

[0323] N-ε26-(19-carboxynonadecanoyl)[Aib8, Arg34]GLP-1-(7-37),

[0324] N-ε26-(4-{[N-(2-carboxyethyl)-N-(15-carboxypentadecanoyl)amino]methyl}benzoyl[Arg34]GLP-1-(7-37),

[0325] N-ε26-[2-(2-[2-(2-[2-(2-[4-(17-carboxyheptadecanoylamino)-4(S)-carboxybutyrylamino]ethoxy)ethoxy]acetylamino)ethoxy]ethoxy)acetyl][Aib8, Arg34]GLP-l-(7-37),

[0326] N-ε26-[2-(2-[2-(2-[2-(2-[4-(19-carboxynonadecanoylamino)-4(S)-carboxybutyrylamino]ethoxy)ethoxy]acetylamino)ethoxy]ethoxy)acetyl][Aib8, Arg34]GLP-1-(7-37),

[0327] N-ε26-[2-(2-[2-(2-[2-(2-[4-(17-carboxyheptadecanoylamino)-4(S)-carboxybutyrylamino]ethoxy)ethoxy]acetylamino)ethoxy]ethoxy)acetyl][3-(4-imidazolyl)propionyl 7,Arg34]GLP-1-(7-37),

[0328] N-ε26-[2-(2-[2-(2-[2-(2-[4-(17-carboxyheptadecanoylamino)-carboxymethyl-amino)acetylamino]ethoxy)ethoxy]acetylamino)ethoxy]ethoxy)acetyl][Aib8, Arg34]GLP-1-(7-37),

[0329] N-ε26-[2-(2-[2-(2-[2-(2-[4-(17-carboxyheptadecanoylamino)-3(S)-sulfopropionylamino]ethoxy)ethoxy]acetylamino)ethoxy]ethoxy)acetyl][Aib8, Arg34]GLP-1-(7-37),

[0330] N-ε26-[2-(2-[2-(2-[2-(2-[4-(17-carboxyheptadecanoylamino)-4(S)-carboxybutyrylamino]ethoxy)ethoxy]acetylamino)ethoxy]ethoxy)acetyl][Gly8,Arg34]GLP-1-(7-37),

[0331] N-ε26-[2-(2-[2-(2-[2-(2-[4-(17-carboxyheptadecanoylamino)-4(S)-carboxybutyrylamino]ethoxy)ethoxy]acetylamino)ethoxy]ethoxy)acetyl][Aib8, Arg34]GLP-1-(7-37)-amide,

[0332] N-ε26-[2-(2-[2-(2-[2-(2-[4-(17-carboxyheptadecanoylamino)-4(S)-carboxybutyrylamino]ethoxy)ethoxy]acetylamino)ethoxy]ethoxy)acetyl][Aib8, Arg34, Pro37]GLP-1-(7-37)amide,

[0333] N-ε26-{2-(2-(2-(2-[2-(2-(4-pentadecanoylamino-4-carboxybutyrylamino)ethoxy)ethoxy]acetyl)ethoxy)ethoxy)acetyl)}[Aib8, Lys26, Arg34]GLP-1(7-37)-OH,

[0334] N-ε26-[2-(2-[2-(2-[2-(2-[4-{[N-(2-carboxyethyl-N-(17-carboxyheptadecanoyl)amino]methyl}benzoyl)amino]ethoxy)ethoxy]acetylamino)ethoxy]ethoxy)acetyl][Aib8,Arg34]GLP-1(7-37),

[0335] N-α7-formyl-N-ε26-[2-(2-[2-(2-[2-(2-[4-(17-carboxyheptadecanoylamino)-4(S)-carboxybutyrylamino]ethoxy)ethoxy]acetylamino)ethoxy]ethoxy)acetyl][Arg34]GLP-1-(7-37),

[0336] N-ε26-[2-(2-[2-(2-[2-(2-[4-(17-carboxyheptadecanoylamino)-4(S)-carboxybutyrylamino]ethoxy)ethoxy]acetylamino)ethoxy]ethoxy)acetyl][Aib8, Glu22, Arg34]GLP-1-(7-37),

[0337] N-ε26{3-[2-(2-{2-[2-(2-{2-[2-(2-[4-(15-(N-((S)-1,3-dicarboxypropyl)carbamoyl)pentadecanoylamino)-(S)-4-carboxybutyrylamino]ethoxy)ethoxy]ethoxy}ethoxy)ethoxy]ethoxy}ethoxy)ethoxy]propionyl}[Aib8, Arg34]GLP-1-(7-37),

[0338] N-ε26-[2-(2-[2-(2-[2-(2-[4-{[N-(2-carboxyethyl)-N-(17-carboxyheptadecanoyl)amino]methyl}benzoyl)amino]-(4-(S)-carboxybutyrylamino)ethoxy)ethoxy]acetylamino)ethoxy]ethoxy)acetyl][Aib8, Arg34]GLP-l(7-37),

[0339] N-ε26-{(S)-4-carboxy-4-((S)-4-carboxy-4-((S)-4-carboxy-4-((S)-4-carboxy-4-(19-carboxynonadecanoylamino)butyrylamino)butyrylamino)butyrylamino}[Aib8, Arg34]GLP-1-(7-37),

[0340] N-ε26-4-(17-carboxyheptadecanoylamino)-4(S)-carboxybutyryl [Aib8, Arg34]GLP-1-(7-37),

[0341] N-ε26-{3-[2-(2-{2-[2-(2-{2-[2-(2-[4-(17-carboxyheptadecanoylamino)-4(S)-carboxybutyrylamino]ethoxy)ethoxy]ethoxy}ethoxy)ethoxy]ethoxy}ethoxy)ethoxy]propionyl}[Aib8, Arg34]GLP-1-(7-37),

[0342] N-ε26-{2-(2-(2-(2-[2-(2-(4-(17-carboxyheptadecanoylamino)-4-carboxybutyrylamino)ethoxy)ethoxy]acetyl)ethoxy)ethoxy)acetyl)}[Aib8, 22, 27, 30, 35, Arg34, Pro37, Lys26]GLP-1(7-37)amide,

[0343] N-ε26-[2-(2-[2-[4-(21-carboxyuneicosanoylamino)-4(S)-carboxybutyrylamino]ethoxy]ethoxy)acetyl][Aib8, Arg34]GLP-1-(7-37) and

[0344] N-ε26-[2-(2-[2-(2-[2-(2-[4-(21-carboxyuneicosanoylamino)-4(S)-carboxybutyrylamino]ethoxy)ethoxy]acetylamino)ethoxy]ethoxy)acetyl][Aib8, Arg34]GLP-1-(7-37) It is a structural analogue of semaglutide selected from the group consisting of:

[0345] The therapeutic polypeptide may be a GIP analog. International Publication Nos. WO 2013 / 164483, WO 2014 / 192284, and WO 2011 / 119657, which are incorporated herein by reference, describe certain GIP analogs as exhibiting GIP and GLP-1 activity. Additional GIP analogs (i.e., agonists of the GLP-1 receptor and the GIP receptor) that exhibit GIP and GLP-1-like activity include those listed in International Publication No. WO 2016 / 111971, which is incorporated herein by reference. The therapeutic polypeptide may be any of these polypeptides. This includes tirzepatide and structurally related analogs thereof.

[0346] Thus, GIP-GLP-1 coagonist compounds for use in the present invention include those of formula III: YX1EGTFTSDYSIX2LDKIAQKAX3VQWLIAGGPSSGAPPPS (Formula III)

[0347] wherein X1 is Aib, X2 is Aib, and K at position 20 is ([2-(2-aminoethoxy)ethoxy]acetyl)2-(γGlu) a -CO-(CH2) b -COH, where a is 1 to 2 and b is 10 to 20, is attached to the ε-amino group of the K side chain; X is Phe or 1-Nal; and the C-terminal amino acid is optionally amidated as a C-terminal primary amide or a pharmaceutically acceptable salt thereof. Includes:

[0348] "Aib" is α-aminoisobutyric acid and "1-Nal" is 1-naphthylalanine.

[0349] In certain embodiments, b is 14 to 18, preferably 16 to 18, or 18.

[0350] In certain embodiments, the C-terminal amino acid is amidated as a C-terminal primary amide.

[0351] Tirzepatide is an example of an incretin peptide analog. Incretins, such as GIP, are a group of metabolic hormones that stimulate a decrease in blood glucose levels. Incretins are released after a meal and increase the secretion of insulin, which is released from the pancreatic beta cells of the islets of Langerhans by a blood glucose-dependent mechanism. Tirzepatide may be considered a dual agonist for the glucose-dependent insulinotropic polypeptide (GIP) receptor and the glucagon-like peptide-1 (GLP-1) receptor. Tirzepatide is marketed under the trade name Mounjaro and has the structure shown in Formula IV and Figure 16C.

[0352] [ka]

[0353] Tirzepatide is (2S)-2-[[20-[[(5S)-6-[[(2S,3S)-1-[[(2S)-1-[[(2S)-5-amino-1-[[(2S)-6-amino-1-[[(2S)-1-[[(2S)-1-[[(2S)-1-[[(2S)-5-amino-1-[[(2S)-1-[[(2S)-1-[[(2S,3S)-1-[[(2S)-1-[[2-[[2-[[(2S)-2-[[(2S)-1-[[(2S)-1-[[2-[[(2S)-1-[( 2S)-2-[(2S)-2-[(2S)-2-[[(2S)-1-amino-3-hydroxy-1-oxopropan-2-yl]carbamoyl]pyrrolidine-1-carbonyl]pyrrolidine-1-carbonyl]pyrrolidin-1-yl]-1-oxopropan-2-yl]amino]-2-oxoethyl]amino]-3-hydroxy-1-oxopropan-2-yl]amino]-3-hydroxy-1-oxopropan-2-yl]carbamoyl]pyrrolidin-1-yl]-2 -oxoethyl]amino]-2-oxoethyl]amino]-1-oxopropan-2-yl]amino]-3-methyl-1-oxopentan-2-yl]amino]-4-methyl-1-oxopentan-2-yl]amino]-3-(1H-indol-3-yl)-1-oxopropan-2-yl]amino]-1,5-dioxopentan-2-yl]amino]-3-methyl-1-oxobutan-2-yl]amino]-1-oxo-3-phenylpropan-2-yl]amino ]-1-oxopropan-2-yl]amino]-1-oxohexan-2-yl]amino]-1,5-dioxopentan-2-yl]amino]-1-oxopropan-2-yl]amino]-3-methyl-1-oxopentan-2-yl]amino]-5-[[(2S)-2-[[(2S)-2-[[2-[[(2S,3S)-2-[[(2S)-2-[[(2S)-2-[[(2S)-2-[[(2S,3R)-2-[[(2S)-2-[[(2S,3R)-2-[[2-[[(2S)-2-[[2-[[(2S)-2-amino-3-(4-hydroxyphenyl)propanoyl]amino]-2-methylpropanoyl]amino]-4-carboxybutanoyl]amino]acetyl]amino]-3-hydroxybutanoyl]amino]-3-phenylpropanoyl]amino]-3-hydroxybutanoyl]amino]-3-hydroxypropanoyl]amino]-3-carboxypropanoyl]amino]-3-(4-hydroxyphenyl)propanoyl It is also known as [2-[2-[2-[2-[2-(carboxymethoxy)ethoxy]ethylamino]-2-oxoethoxy]ethoxy]ethylamino]-5-oxopentanoic acid.

[0354] WO 2019 / 125929 and WO 2019 / 125938, both incorporated herein by reference, describe certain incretin mimetics as exhibiting GIP, GLP-1, and glucagon-like activity (i.e., agonists of the GLP-1 receptor, GIP receptor, and glucagon receptor). This includes retatortide (LY3437943) and its structurally related analogs. Retatortide may be considered a structural analog of tirzepatide, and vice versa. The therapeutic polypeptide may be any of these polypeptides.

[0355] Thus, GIP-GLP-1-glucagon triple co-agonist compounds (incretin analogs) for use in the present invention include those of Formula V: YX2QGTFTSDYSIX 13 LDKX 17 AX 19 X 20 AFIEYLLX 28 X 29 GPSSX 34 APPPS (Formula V)

[0356] During the ceremony, X2 is Aib, X 13 is L or αMeL, X 17 is any amino acid having a functional group available for conjugation, the functional group being attached to a C16-C22 fatty acid, optionally via a linker; X 19 is Q or A, X 20 is Aib, αMeK (α-methyl-L-lysine), Q or H, X 28 is E or A, X 29 is G or Aib, X 34 is G or Aib, and the C-terminal amino acid is optionally amidated; Includes:

[0357] In certain embodiments, position X 17 The amino acid having a functional group available for conjugation in is selected from the group consisting of K, C, E and D, preferably K.

[0358] In certain embodiments, the linker comprises 1 to 4 amino acids, preferably Glu or γGlu. In other embodiments, the linker comprises H-{NH—CH—CH—[O—CH—CH] m -O-(CH2) p -CO} n Further included are structures of -OH, wherein m is an integer of any one of 1 to 12, n is an integer of any one of 1 to 12, and p is 1 or 2.

[0359] In certain embodiments, the linker further comprises one to four (2-[2-(2-aminoethoxy)ethoxy]acetyl) (ie, AEEA) moieties.

[0360] In certain specific embodiments, X17 has the following structure: (2-[2-(2-aminoethoxy)ethoxy]acetyl)-(γGlu) b -CO-(CH2) c K chemically modified by attachment to the ε-amino group of the K side chain with —COH, where a is 0, 1, or 2, b is 1 or 2, and c is an integer between 16 and 18.

[0361] In certain specific embodiments, the triple agonist incretin analog is retatortide (LY3437943), the structure of which is shown in Formula VI and Figure 16D. YAibQGTFTSDYSIαMeLLDK K AQAibAFIEYLLEGGPSSGAPPPS-CONH2 (Formula VI)

[0362] During the ceremony, K is connected to C via a (AEEA)γGlu linker 20 Attached to an aliphatic diacid moiety.

[0363] Accordingly, the present invention further provides a solid formulation adapted for oral delivery to the stomach of a human or non-human animal subject, said composition comprising: (i) Dulaglutide, exenatide, liraglutide, semaglutide, tirzepatide, albiglutide, lixisenatide, polyethylene glycol oxenatide, retatortide (LY3437943), cotadutide, taspoglutide, langrenatide, veinaglutide, efpegrenatide, LY3502970, LY3537031, LY3493269, HM12525A / JNJ-64565111, MOD6030 / 1, SAR425899, MEDI0382, MK8521, ZP2929 / BI a polypeptide therapeutic selected from the group consisting of 456906, NN9709 / NNC0090-2746 / MAR709 / RG7697 / R06811135, SAR441255, C2816, ZP3022, NNC9204-1177 (NN9277), LY3305677, JNJ-54728518, LY2944876 / TT-401, CPD86, SAR438335, ZP-I-98, ZP-DI-70, HM15211, NN9423 / MAR423, PB-719, and DD01; (ii) an alginate oligomer; Including, The solid dosage form does not have a coating that provides substantial protection from the gastric environment in addition to the polypeptide therapeutic agent and alginate oligomer.

[0364] The above-described dosage forms may further comprise a gastrointestinal permeation enhancer as described herein.

[0365] In certain embodiments, the solid formulation (composition) is in the form of a tablet, pill, or powder- or granule-filled capsule. In certain embodiments, the solid formulation has dimensions that allow it to be swallowed by the subject to be treated. In certain embodiments, the solid formulation begins to dissolve or disperse in the gastric environment of the subject to be treated, for example, in the gastric fluid of the subject to be treated, or in a surrogate thereof, immediately after entering the stomach. The above detailed discussion of the dosage forms of the present invention applies mutatis mutandis to this aspect of the present invention.

[0366] Therapeutic polypeptides may be used in the methods described herein or provided in the compositions or dosage forms described herein in their free or pharmaceutically acceptable salt form. For example, corresponding pharmaceutically acceptable salts may be formed by protonation of an atom having a lone pair of electrons susceptible to protonation, such as an amino group, with an inorganic or organic acid, or as salts of carboxylic acid groups with physiologically acceptable cations, as is well known in the art. Exemplary base addition salts include, for example, alkali metal salts such as sodium or potassium salts; alkaline earth metal salts such as calcium or magnesium salts; zinc salts; ammonium salts; aliphatic amine salts such as trimethylamine, triethylamine, dicyclohexylamine, ethanolamine, diethanolamine, triethanolamine, procaine salts, meglumine salts, ethylenediamine salts, or choline salts; aralkylamine salts such as N,N-dibenzylethylenediamine salts, benzathine salts, benethamine salts; heterocyclic aromatic amine salts such as pyridine salts, picoline salts, quinoline salts, or isoquinoline salts; quaternary ammonium salts such as tetramethylammonium salts, tetraethylammonium salts, benzyltrimethylammonium salts, benzyltriethylammonium salts, benzyltributylammonium salts, methyltrioctylammonium salts, or tetrabutylammonium salts; and basic amino acid salts such as arginine salts, lysine salts, or histidine salts.Exemplary acid addition salts are, for example, inorganic acid salts such as hydrochloride, hydrobromide, hydroiodide, sulfate, nitrate, phosphate (e.g., phosphate, hydrogen phosphate, or dihydrogen phosphate), carbonate, bicarbonate, or perchlorate; acetate, propionate, butyrate, pentanoate, hexanoate, heptanoate, octanoate, cyclopentanepropionate, decanoate, undecanoate, oleate, stearate, lactate, maleate, oxalate, fumarate, tartrate, malate, citrate, succinate, glycolate, dihydroxybenzo ... Organic acid salts such as cotinate, benzoate, salicylate, ascorbate or pamoate (embonate); sulfonate salts such as methanesulfonate (mesylate), ethanesulfonate (esylate), 2-hydroxyethanesulfonate (isethionate), benzenesulfonate (besylate), p-toluenesulfonate (tosylate), 2-naphthalenesulfonate (napsylate), 3-phenylsulfonate or camphorsulfonate; and acidic amino acid salts such as aspartate or glutamate.

[0367] The uses, methods, and compositions of the invention may comprise several / multiple therapeutic polypeptides. Thus, the methods, uses, and compositions of the invention may comprise at least one, or one or more therapeutic polypeptides (e.g., at least two, three, or four therapeutic polypeptides, or two, three, four, or more therapeutic polypeptides). These may be structural or functional analogs of each other, or therapeutic polypeptides from different structural or functional classes. References herein to "a" or "the" polypeptide should be interpreted as covering such embodiments, unless otherwise indicated. In particular, compositions described herein as "consisting of" a set of components of which a "therapeutic polypeptide" is a part include a combination of therapeutic polypeptides as said components.

[0368] A gastrointestinal permeation enhancer (also known as a mucosal permeability enhancer or simply permeability enhancer), more specifically a gastrointestinal epithelial barrier (epithelial) permeability enhancer, is a compound that facilitates the movement of other compounds from the luminal (apical) side of the gastrointestinal epithelial layer to the basolateral side of said layer. Permeability enhancers may have a paracellular or transcellular mechanism of action. Permeability enhancers may also have properties that improve the ability of therapeutic polypeptides to penetrate mucus, although this is not required. Without wishing to be bound by theory, alginate oligomers provide such activity, and therefore, in certain embodiments, the gastrointestinal permeation enhancer is not an agent that affects the structure or viscosity of mucus.

[0369] The permeability enhancer is not an alginate oligomer. In certain embodiments, the permeability enhancer is not or does not include a polypeptide or peptide.

[0370] In certain embodiments, the gastrointestinal permeation enhancer may have pH buffering properties in a gastric environment (simulated or natural), such that a dosage form, e.g., a solid formulation containing the gastrointestinal permeation enhancer, when placed in a gastric environment (simulated or natural) causes the pH of its immediate vicinity to be closer to neutral than the gastric environment in which it is placed.

[0371] The permeability enhancer is C 8-20 Alkanoylcarnitine (preferably lauroylcarnitine, myristoylcarnitine or palmitoylcarnitine, for example, lauroylcarnitine chloride, myristoylcarnitine chloride or palmitoylcarnitine chloride), salicylic acid (preferably a salicylate, for example, sodium salicylate), salicylic acid derivatives (for example, 3-methoxysalicylic acid, 5-methoxysalicylic acid or homovanillic acid), C 8-20 Alkanoic acid (preferably C 8-20Alkanoates, more preferably caprates, caprylates, myristates, palmitates or stearates, such as sodium caprate, sodium caprylate, sodium myristate, sodium palmitate or sodium stearate, citric acid (preferably a citrate, such as sodium citrate), tartaric acid (preferably a tartrate), fatty acylated amino acids (such as sodium lauroylalanine, as described in U.S. Patent Application Publication No. 2014 / 0056953 A1, which is incorporated herein by reference), N-Dodecanoyl-L-alanine, Sodium Lauroyl Asparagine, N-Dodecanoyl-L-Asparagine, Sodium Lauroyl Aspartate, N-Dodecanoyl-L-Aspartic Acid, Sodium Lauroyl Cysteine, N-Dodecanoyl-L-Cysteine, Sodium Lauroyl Glutamate, N-Dodecanoyl-L-Glutamate, Sodium Lauroyl Glutamine, N-Dodecanoyl-L-Glutamine, Sodium Lauroyl Glycine, N-Dodecanoyl-L-Glycine, Sodium Lauroyl Histidine, N-Dodecanoyl-L-Histidine Stidine, Sodium Lauroyl Isoleucine, N-Dodecanoyl-L-Isoleucine, Sodium Lauroyl Leucine, N-Dodecanoyl-L-Leucine, Sodium Lauroyl Methionine, N-Dodecanoyl-L-Methionine, Sodium Lauroyl Phenylalanine, N-Dodecanoyl-L-Phenylalanine, Sodium Lauroyl Proline, N-Dodecanoyl-L-Proline, Sodium Lauroyl Serine, N-Dodecanoyl-L-Serine, Sodium Lauroyl Threonine, N-Dodecanoyl-L-Threonine, Sodium Lauroyl Triphosphate tryptophan, N-dodecanoyl-L-tryptophan, sodium lauroyl tyrosine, N-dodecanoyl-L-tyrosine, sodium lauroyl valine, N-dodecanoyl-L-valine, sodium lauroyl sarcosine, N-dodecanoyl-L-sarcosine, sodium caproyl alanine, N-decanoyl-L-alanine, sodium caproyl asparagine, N-decanoyl-L-asparagine, sodium caproyl aspartic acid, N-decanoyl-L-aspartic acid, sodium caproyl cysteine, N-decanoyl-L-cysteine,Sodium Caproyl Glutamate, N-Decanoyl-L-Glutamate, Sodium Caproyl Glutamine, N-Decanoyl-L-Glutamine, Sodium Caproyl Glycine, N-Decanoyl-L-Glycine, Sodium Caproyl Histidine, N-Decanoyl-L-Histidine, Sodium Caproyl Isoleucine, N-Decanoyl-L-Isoleucine, Sodium Caproyl Leucine, N-Decanoyl-L-Leucine, Sodium Caproyl Methionine, N-Decanoyl-L-Methionine, Sodium Caproyl Phenylalanine, N-Deca N-decanoyl-L-phenylalanine, sodium caproyl prolinate, N-decanoyl-L-proline, sodium caproyl serine, N-decanoyl-L-serine, sodium caproyl threonine, N-decanoyl-L-threonine, sodium caproyl tryptophan, N-decanoyl-L-tryptophan, sodium caproyl tyrosine, N-decanoyl-L-tyrosine, sodium caproyl valine, N-decanoyl-L-valine, sodium caproyl sarcosine, N-decanoyl-L-sarcosine, sodium oleoyl sarcosine, sodium Sodium N-decyl leucine, sodium stearoyl glutamine (e.g., Amisoft HS-11P), sodium myristoyl glutamine (e.g., Amisoft MS-11), sodium lauroyl glutamine (e.g., Amisoft LS-11), sodium cocoyl glutamine (e.g., Amisoft CS-11), sodium cocoyl glycine (e.g., Amisoft GCS-11), sodium N-decyl leucine, sodium cocoyl glycine, sodium cocoyl glutamine, sodium lauroyl alanine, N-dodeca N-dodecanoyl-L-alanine, sodium lauroyl asparagine, N-dodecanoyl-L-asparagine, sodium lauroyl aspartate, N-dodecanoyl-L-aspartate, sodium lauroyl cysteine, N-dodecanoyl-L-cysteine, sodium lauroyl glutamate, N-dodecanoyl-L-glutamate, sodium lauroyl glutamine, N-dodecanoyl-L-glutamine, sodium lauroyl glycine, N-dodecanoyl-L-glycine, sodium lauroyl histidine, N-dodecanoyl-L-histidine,Sodium lauroyl isoleucine, N-dodecanoyl-L-isoleucine, sodium lauroyl leucine, N-dodecanoyl-L-leucine, sodium lauroyl methionine, N-dodecanoyl-L-methionine, sodium lauroyl phenylalanine, N-dodecanoyl-L-phenylalanine, sodium lauroyl proline, N-dodecanoyl-L-proline, sodium lauroyl serine, N-dodecanoyl-L-serine, sodium lauroyl threonine, N-dodecanoyl-L-threonine, sodium lauroyl tryptophan, N-Dodecanoyl-L-tryptophan, Sodium Lauroyl Tyrosine, N-Dodecanoyl-L-Tyrosine, Sodium Lauroyl Valine, N-Dodecanoyl-L-Valine, N-Dodecanoyl-L-Sarcosine, Sodium Caproyl Alanine, N-Decanoyl-L-Alanine, Sodium Caproyl Asparagine, N-Decanoyl-L-Asparagine, Sodium Caproyl Aspartate, N-Decanoyl-L-Aspartate, Sodium Caproyl Cysteine, N-Decanoyl-L-Cysteine, Sodium Caproyl Glutamate, N-Deca N-decanoyl-L-glutamic acid, sodium caproyl glutamine, N-decanoyl-L-glutamine, sodium caproyl glycine, N-decanoyl-L-glycine, sodium caproyl histidine, N-decanoyl-L-histidine, sodium caproyl isoleucine, N-decanoyl-L-isoleucine, sodium caproyl leucine, N-decanoyl-L-leucine, leucine, sodium caproyl methionine, N-decanoyl-L-methionine, sodium caproyl phenylalanine, N-decanoyl-L-phenylalanine, sodium sodium caproylproline, N-decanoyl-L-proline, sodium caproylserine, N-decanoyl-L-serine, sodium caproylthreonine, N-decanoyl-L-threonine, sodium caproyltryptophan, N-decanoyl-L-tryptophan, sodium caproyltyrosine, N-decanoyl-L-tyrosine, sodium caproylvaline, N-decanoyl-L-valine, sodium caproylsarcosine, sodium oleoylsarcosine and pharmaceutically acceptable salts of any of the above compounds, or, for example, C,8-20 Alkanoyl sarcosine (e.g., lauroyl sarcosine such as sodium lauroyl sarcosine) or C 8-20 any of the 20 standard proteinogenic α-amino acids acylated with an alkanoic acid, including but not limited to, any of the fatty acid acylated amino acids), alkyl sugars (e.g., C 1-20 Alkyl sugars, such as C such as Multitrope™ 1620-LQ-(MV) 8-10 Alkyl polysaccharides, such as n-octyl-β-D-glucopyranoside, n-dodecyl-β-D-maltoside, n-tetradecyl-β-D-maltoside, tridecyl-β-D-maltoside, sucrose laurate, sucrose stearate, sucrose myristate, sucrose palmitate, sucrose cocoate, sucrose monododecanoate, sucrose monotridecanoate, sucrose monotetradecanoate, coco-glucoside, or any of the alkyl sugars described in U.S. Pat. No. 5,661,130 or WO 2012 / 112319, which are incorporated herein by reference), cyclodextrins (e.g., α-cyclodextrin, β-cyclodextrin, γ-cyclodextrin, methyl- β-cyclodextrin, hydroxypropyl β-cyclodextrin or sulfobutylether β-cyclodextrin), N-[8-(2-hydroxybenzoyl)amino]caprylic acid (preferably N-[8-(2-hydroxybenzoyl)amino]caprylate, more preferably sodium N-[8-(2-hydroxybenzoyl)amino]caprylate, also known as "SNAC"), N-[8-(2-hydroxybenzoyl)amino]caprylate salt derivatives (preferably sodium N-[8-(2-hydroxybenzoyl)amino]caprylate derivatives), calcium chelating compounds (e.g., ethylenediaminetetraacetic acid (EDTA), ethylene glycol tetraacetic acid (EGTA), Cremophor EL (also known as "Kolliphor EL"), CAS 911-100-1, 200-21 ...310-1, 200-310-1, 200-410-1, 200-410-1, 200-510-1, 200-510-1, 200-610-1, 200-710-1, 200-810-2, 200-910-2, 200-1000-3, 200-110-1, 200-210-2, 200-310-2, 200-410-3, 200-510-1, 200-610-1, 200-710-1, 200-810-2, 200-910-3, 200-110-1, 200-120-1, 2 No. 61791-12-6), chitosan, N,N,N-trimethylchitosan, benzalkonium chloride, bestatin, cetylpyridinium chloride, cetyltrimethylammonium bromide, C2-20 Alkanols (e.g., ethanol, decanol, lauryl alcohol, myristyl alcohol, or palmityl alcohol), C 8-20 Alkenols (e.g., oleyl alcohol), C 8-20 Alkenoic acids (e.g., oleic acid), dextran sulfate, diethylene glycol monoethyl ether (Transcutol), 1-dodecylazacycloheptan-2-one (Azone®), caprylocaproyl polyoxylglycerides (e.g., caprylocaproyl polyoxyl-8 glycerides, available as, for example, Labrasol® or ACCONON® MC8-2), ethyl caprylate, glyceryl monolaurate, lysophosphatidylcholine, menthol, C 8-20 Alkylamines, C 8-20 alkenylamines (e.g., oleylamine), phosphatidylcholine, poloxamers, polyethylene glycol monolaurate, polyoxyethylene, polypropylene glycol monolaurate, polysorbates (e.g., polysorbate 20 or polysorbate 80), cholic acid (preferably cholate salts, e.g., sodium cholate), deoxycholate salts (e.g., sodium deoxycholate), chenodeoxycholate salts (e.g., sodium chenodeoxycholate), sodium glycocholate, sodium glycodeoxycholate, sodium lauryl sulfate (SDS), sodium decyl sulfate, sodium octyl sulfate, sodium laureth sulfate, N-lauryl sarcosinate, decyltrimethylammonium bromide, benzyldimethyldodecylammonium chloride, myristyltrimethylammonium chloride, dodecylpyridinium chloride, decyldimethylammoniopropanesulfonate, myristyldimethylammoniopropanesulfonate, palmityldimethylammoniopropanesulfonate , ChemBetaine CAS, ChemBetaine Oleyl, Nonylphenoxypolyoxyethylene, Polyoxyethylene Sorbitan Monolaurate, Polyoxyethylene Sorbitan Monopalmitate, Sorbitan Monooleate, Triton X-100, Hexanoic Acid, Heptanoic Acid, Methyl Laurate, Isopropyl Myristate, Isopropyl Palmitate, Methyl Palmitate, Diethyl Sebacate, Sodium Oleate, Urea, Laurylamine, Caprolactam, Methylpyrrolidone, Octylpyrrolidone, Methylpiperazine, Phenylpiperazine, Carbopol 934P, Glycyrrhetinic Acid, Bromelain, Pinene Oxide, Limonene, Cineole, Octyldodecanol, Fenchon, Menthone, Examples of permeability enhancers include trimethoxypropylenemethylbenzene, macrogol-15-hydroxystearate, taurocholate (e.g., sodium taurocholate), taurodeoxycholate (e.g., sodium taurodeoxycholate), sulfoxides (e.g., decylmethyl sulfoxide or dimethyl sulfoxide), cyclopentadecalactone, 8-(N-2-hydroxy-5-chlorobenzoyl)-amino-caprylic acid (5-CNAC), N-(10-[2-hydroxybenzoyl]amino)decanoic acid (SNAD), dodecyl-2-N,N-dimethylaminopropionate (DDAIP), D-α-tocopheryl polyethylene glycol-1000 succinate (TPGS), arginine, and pharmaceutically acceptable salts of the foregoing compounds. Mixtures of two or more permeability enhancers, including any of the above-listed permeability enhancers, can also be used.

[0372] Alkyl glycosides, which may also be used as permeability enhancers in the present invention, include octyl-, nonyl-, decyl-, undecyl-, dodecyl-, tridecyl-, tetradecyl-, pentadecyl-, hexadecyl-, heptadecyl-, and octadecyl-α- or β-D-maltosides, -glucosides, or -sucroses; alkyl thiomaltosides, such as heptyl-, octyl-, dodecyl-, tridecyl-, and tetradecyl-β-D-thiomaltoside; alkyl thioglucosides, such as heptyl-, octyl-, dodecyl-, tridecyl-, and tetradecyl-β-D-thiomaltoside; These may include tyl- or octyl 1-thio α-β- or β-D-glucopyranoside, alkylthiosucrose, alkylmaltotriosides, long-chain aliphatic carbonic acid amides of sucrose β-amino-alkyl ethers, derivatives of palatinose and isomaltamine linked to the alkyl chain by an amide bond, derivatives of isomaltamine linked to the alkyl chain by a urea bond, long-chain aliphatic carbonic acid ureides of sucrose β-amino-alkyl ethers, and long-chain aliphatic carbonic acid amides of sucrose β-amino-alkyl ethers.

[0373] In another further embodiment, the permeability enhancer may be of formula (VII):

[0374] [ka]

[0375] During the ceremony, R 1 , R 2 , R 3 and R 4 are each independently hydrogen, -OH, or -NR 6 R 7 , halogen (e.g., -F, -Cl, -Br, or -I), C 1-4 Alkyl or C 1-4 alkoxy; R 5 is a substituted or unsubstituted C 2-16 Alkylene, substituted or unsubstituted C 2-16 Alkenylene, substituted or unsubstituted C 1-12Alkyl(arylene) [e.g., substituted or unsubstituted C 1-12 alkyl(phenylene)], or substituted or unsubstituted aryl (C 1-12 alkylene) [e.g., substituted or unsubstituted phenyl (C 1-12 alkylene)], and R 6 and R 7 are each independently hydrogen, oxygen, -OH or C 1-4 alkyl, or a pharmaceutically acceptable salt or solvate thereof, in particular a disodium salt, an alcohol solvate (e.g., a methanol solvate, an ethanol solvate, a propanol solvate, or a propylene glycol solvate, or such a solvate of the disodium salt, in particular an ethanol solvate or an ethanol solvate of the disodium salt), or a hydrate thereof (e.g., a monohydrate of the disodium salt). The above-mentioned "substituted" groups contained in formula (V) are preferably halogen (e.g., -F, -Cl, -Br, or -I), -OH, C 1-4 Alkyl or C 1-4 The compound may be substituted with one or more (eg, 1, 2, or 3) substituents independently selected from alkoxy.

[0376] For example, WO 00 / 59863 describes such compounds and methods for their preparation, which are incorporated herein by reference. Thus, the permeability enhancer may also be a "delivery agent" described in WO 00 / 59863. Preferred examples of compounds of formula (V) include N-(5-chlorosalicyloyl)-8-aminocaprylic acid, N-(10-[2-hydroxybenzoyl]amino)decanoic acid, N-(8-[2-hydroxybenzoyl]amino)caprylic acid, the monosodium or disodium salt of any one of the foregoing compounds, an ethanol solvate of the sodium salt (e.g., the monosodium or disodium salt) of any one of the foregoing compounds, the monohydrate of the sodium salt (e.g., the monosodium or disodium salt) of any one of the foregoing compounds, and any combination thereof. A particularly preferred compound of formula (I) is the disodium salt of N-(5-chlorosalicyloyl)-8-aminocaprylic acid or its monohydrate.

[0377] It is particularly preferred if the penetration enhancer is selected from sodium caprate, sodium caprylate, a mixture of sodium caprate and sodium caprylate, SNAC, sucrose laurate, Labrasol and polysorbates.

[0378] Additionally, the permeability enhancer may be a salt of a medium chain fatty acid. The salt of a medium chain fatty acid is preferably C 4-18 saturated fatty acids, preferably C═C with one, two or three C═C double bonds as appropriate 4-18 Linear or branched alkanoic acids, more preferably C═C ... 6-16 Linear or branched alkanoic acids, more preferably C═C, optionally with one, two or three C═C double bonds 6-14 The salts of linear or branched alkanoic acids are preferably C 4-18 Linear or branched alkanoic acids, more preferably C 6-16 Linear or branched alkanoic acids, more preferably C 6-14The salt of a medium-chain fatty acid is preferably selected from the group consisting of valeric acid, caproic acid, enanthic acid, caprylic acid, pelargonic acid, capric acid, undecylic acid, lauric acid, tridecylic acid, myristic acid, pentadecylic acid, palmitic acid, margaric acid, and / or stearic acid. More preferably, the salt of a medium-chain fatty acid is a salt of capric acid.

[0379] The salt of the medium-chain fatty acid is more preferably a sodium salt or a potassium salt. The salt of the medium-chain fatty acid is even more preferably a salt of capric acid. Capric acid is sometimes called decanoic acid (CH3(CH2)8COOH). A preferred salt of capric acid is sodium caprate (i.e., CH3(CH2)8COONa).

[0380] In certain embodiments, the paracellular permeability enhancer may be a chelating agent, such as EDTA, citric acid, and salicylate. Transcellular enhancers include medium-chain fatty acids (oleic acid, lauric acid, acylcarnitines, acylcholines, acylated amino acids), bile salts (e.g., sodium glycocholate, sodium deoxycholate, sodium taurocholate, sodium dihydrofusidate, sodium glycodihydrofusidate, sodium glycolate), ionic surfactants (e.g., sodium lauryl sulfate and dioctyl sodium sulfosuccinate), nonionic surfactants (e.g., sucrose fatty acid esters, lactose fatty acid esters, polysorbates, polyoxyethylene-8 lauryl ether (C 12 E8), Tween 80, sucrose laurate, macrogol-8 glyceride, lauroylcarnitine chloride, caprylic acid, sodium caprylate, sodium caprate (sodium decanoate, C 10), sodium cholate, choline geranate, 5-CNAC (N-(5-chlorosalicyloyl)-8-aminocaprylic acid), chitosan, chitosan glutamate, N-sulfant-N,O-carboxymethylchitosan (SNOCC), N-trimethylated chitosan chloride (TMC), ethanol, salicylate, penetratin, Zonula occludens toxin (Zot), polycarbophil-cysteine ​​complex (PCP-Cys).

[0381] More specifically, the gastrointestinal permeation enhancer may be any from the following non-exhaustive list:

[0382] Sodium N-(8-[2-hydroxybenzoyl]amino)caprylate (SNAC), decanoic acid and its salts (C10), octanoic acid and its salts (C8), dodecanoic acid and its salts (C12), icosanoic acid and its salts (C20), palmitoylcarnitine chloride, lauroylcarnitine, sodium salicylate, 3-methoxysalicylic acid, 5-methoxysalicylic acid, homovanillic acid, sodium lauroylalanine, N-dodecanoyl-L-glutamic acid, sodium caproylsarcosine, sodium stearoylglutamate (e.g., Amisoft 20 HS-11 P), C8-10 alkyl polysaccharides (e.g., Multitrope™ 1620-LQ-MV), N-dodecyl-β-D-maltoside, ethylene glycol tetraacetic acid (EGTA), polyoxyl-35 castor oil (also known as Cremophor EL and Koriphor EL, CAS number 61791-12-6);Trimethylchitosan, N,N,N-trimethylchitosan, benzalkonium chloride, cetylpyridinium chloride, cetyltrimethylammonium bromide, ethanol, oleyl alcohol, diethylene glycol monoethyl ether (Transcutol), 1-dodecylazacyclo-heptan-2-one (Azone®), Labrasol® or Aconon® MC8-2, phosphatidylcholine, sodium cholate, dodecylphosphocholine, rhamnolipid, Carbopol 934P, octylpyrrolidone, sodium taurocholate, 8-(N- 2-Hydroxy-5-chloro-benzoyl)aminocaprylic acid (5-CNAC), N-(10-[2-hydroxybenzoyl]amino)decanoic acid (SNAD), D-α-tocopheryl polyethylene glycol 1000 succinate (TPGS), sodium decanoate (sodium caprate), octyl gallate, sodium octanoate (sodium caprylate), lauryl gallate, dodecyl maltoside (DDM), tetradecyl maltoside (TDM), palmitoyl carnitine, sodium chenodeoxycholate (NaCDC), polyoxyl 15-hydroxystearate (Colifor Important enhancers that may be used alone or in combination in the present invention include HS 15, sodium deoxycholate, polyoxyethylene (10) oleyl ether (Brij 10), sodium tetraglycocholate, polyethylene glycol hexadecyl ether, sodium ursodeoxycholate, polyoxyethylene (10) cetyl ether, sodium glycocholate, Purebright PEG, sodium taurocholate, diethylene glycol monoethyl ether (Transcutol ES), chenodeoxycholic acid, lauroglycol FCC, ethyl gallate, propyl gallate (PG), chitosan, arachidonic acid, SNAC, and C10 (especially the sodium salt form, sodium caprate) are important enhancers that may be used alone or in combination in the present invention;

[0383] In certain embodiments, the delivery agent (penetration enhancer) may be a salt of N-(8-(2-hydroxybenzoyl)amino)caprylic acid.

[0384] The structural formula of N-(8-(2-hydroxybenzoyl)amino)caprylate is shown in formula (VIII) below.

[0385] [ka]

[0386] In some embodiments, the salt of N-(8-(2-hydroxybenzoyl)amino)caprylic acid includes one monovalent cation, two monovalent cations, or one divalent cation. In some embodiments, the salt of N-(8-(2-hydroxybenzoyl)amino)caprylic acid is selected from the group consisting of sodium, potassium, and calcium salts of N-(8-(2-hydroxybenzoyl)amino)caprylic acid. Salts of N-(8-(2-hydroxybenzoyl)amino)caprylic acid may be prepared using, for example, the methods described in WO 96 / 030036, WO 00 / 046182, WO 01 / 092206, or WO 2008 / 028859, which are incorporated herein by reference.

[0387] The salt of N-(8-(2-hydroxybenzoyl)amino)caprylic acid can be crystalline and / or amorphous. In some embodiments, the delivery agent comprises an anhydrate, monohydrate, dihydrate, trihydrate, solvate, or one-third hydrate of a salt of N-(8-(2-hydroxybenzoyl)amino)caprylic acid, as well as combinations thereof. In some embodiments, the delivery agent is a salt of N-(8-(2-hydroxybenzoyl)amino)caprylic acid described in WO 2007 / 121318, incorporated herein by reference.

[0388] In some embodiments, the delivery agent is sodium N-(8-(2-hydroxybenzoyl)amino)caprylate (referred to herein as "SNAC"), also known as sodium 8-(salicyloylamino)octanoate.

[0389] In some embodiments, the amount of the salt of N-(8-(2-hydroxybenzoyl)amino)caprylic acid in a composition or dosage form comprising a therapeutic polypeptide and optionally an alginate oligomer is at least 0.6 mmol, e.g., at least 0.65 mmol, at least 0.7 mmol, at least 0.75 mmol, at least 0.8 mmol, at least 0.8 mmol, at least 0.9 mmol, at least 0.95 mmol, and at least 1 mmol. Any range, which may be comprised of endpoints above, is expressly contemplated. In some embodiments, the amount of the salt of N-(8-(2-hydroxybenzoyl)amino)caprylic acid in the composition is in the range of 0.6 to 2.1 mmol, or 0.6 to 1.9 mmol. In some embodiments, the amount of the salt of N-(8-(2-hydroxybenzoyl)amino)caprylic acid in the composition is in the range of 0.7 to 1.7 mmol, or 0.8 to 1.3 mmol.

[0390] In some embodiments, the amount of salt of N-(8-(2-hydroxybenzoyl)amino)caprylic acid in a composition or dosage form comprising a therapeutic polypeptide and optionally an alginate oligomer is up to 2.1 mmol, e.g., up to 2.1 mmol, up to 2 mmol, up to 1.9 mmol, up to 1.8 mmol, up to 1.7 mmol, up to 1.6 mmol, up to 1.5 mmol, up to 1.4 mmol, up to 1.3 mmol, up to 1.2 mmol, and up to 1.1 mmol. Any range, which may be made up of endpoints of the above values, is expressly contemplated. In some embodiments, the amount of salt of N-(8-(2-hydroxybenzoyl)amino)caprylic acid is 1 mmol, e.g., 1.08 mmol.

[0391] In other embodiments, the amount of salt of N-(8-(2-hydroxybenzoyl)amino)caprylic acid in a composition or dosage form comprising a therapeutic polypeptide and optionally an alginate oligomer is less than 0.6 mmol, e.g., less than 0.55 mmol, less than 0.5 mmol, or less than 0.45 mmol. Any range that may be made up of endpoints above is expressly contemplated.

[0392] In some embodiments, the amount of salt of N-(8-(2-hydroxybenzoyl)amino)caprylic acid in a composition or dosage form comprising a therapeutic polypeptide and optionally an alginate oligomer is less than 50% wt / w, e.g., less than 45% wt / w, less than 40% wt / w, less than 35% wt / w, less than 30% wt / w, less than 25% wt / w, less than 20% wt / w, less than 15% wt / w, or less than 10% wt / w. Any range that may be made up of endpoints of the above values ​​is expressly contemplated.

[0393] In some embodiments, the amount of SNAC in the composition or dosage form is at least 30 mg, e.g., at least 50, 70, 90, 110, 130, 150, or 170 mg. In some embodiments, the amount of SNAC in the composition is at least 175 mg, e.g., an amount selected from the group consisting of at least 200 mg, at least 210 mg, at least 220 mg, at least 230 mg, at least 240 mg, at least 250 mg, at least 260 mg, at least 270 mg, and at least 280 mg. Any range that may be formed from the endpoints recited above is expressly contemplated.

[0394] In some embodiments, the amount of SNAC in a composition or dosage form is in the range of 30 to 200 mg, e.g., 50, 70, 90, 110, 130, 150, 170, or 190 mg to 200 mg, or 50 to 70, 90, 110, 130, 150, 170, 190, or 200 mg. In some embodiments, the amount of SNAC in a composition is in the range of 175 to 575 mg, e.g., 200 to 500 mg or 250 to 400 mg. In some embodiments, the amount of SNAC in a composition is an amount selected from the group consisting of up to 575 mg, e.g., up to 550 mg, up to 525 mg, up to 500 mg, up to 475 mg, up to 450 mg, up to 425 mg, up to 400 mg, up to 375 mg, up to 350 mg, and up to 325 mg. Any range formed by endpoints of the values ​​recited above is expressly contemplated.

[0395] In some embodiments, the amount of SNAC in the composition or dosage form is about 300, 200, 100, 50, or 30 mg. Any range made up of endpoints above is expressly contemplated.

[0396] In some embodiments, the molar ratio between the therapeutic polypeptide (eg, GLP-1 receptor agonist) and the permeability enhancer in the composition or dosage form is less than 10, such as less than 5 or less than 1.

[0397] In some embodiments, the amount of SNAC in a composition or dosage form containing a therapeutic polypeptide and optionally an alginate oligomer is less than 175 mg, e.g., less than 170 mg, 165 mg, 160 mg, 155 mg, 150 mg, 145 mg, 140 mg, 135 mg, 130 mg, 125 mg, 120 mg, 115 mg, 110 mg, 105 mg, 100 mg, 90 mg, 80 mg, 70 mg, 60 mg, 50 mg, or 40 mg. Any range formed by endpoints of the above values ​​is expressly contemplated.

[0398] In these embodiments, the composition or dosage form, e.g., in tablet form, has a weight within the range of 50 to 1000 mg, e.g., 100, 150, 200, 250, 300, 350, 400, 450, 500, 550, 600, 1000 mg, or 100 to 150, 650, 700, 750, 800, 850, 900, 950, 200, 250, 300, 350, 400, 450, 500, 550, 600, 650, 700, 750, 800, 850, 900, 950, or 1000 mg. Any range that can be made up of endpoints containing the above values ​​is expressly contemplated.

[0399] In these embodiments, the composition or dosage form, e.g., in tablet form, has a weight in the range of 175 mg to 1000 mg, e.g., 175-250 mg, 300-500 mg, or 500-900 mg, or e.g., about 200 mg, about 400 mg, or about 700 mg. In some embodiments, the tablet weight is in the range of 200 mg to 1000 mg, e.g., 500-700 mg, or 600-1000 mg, or e.g., about 200 mg, about 400 mg, about 600 mg, or about 800 mg.

[0400] Thus, the present invention further provides a dosage form adapted for delivery to the stomach of a human or non-human animal subject, said dosage form comprising: (i) a polypeptide therapeutic; and (ii) an alginate oligomer; (iii) EDTA, citric acid, salicylate, oleic acid, lauric acid, acylcarnitine, acylcholine and acylated amino acids, sodium glycocholate, sodium deoxycholate, sodium taurocholate, sodium dihydrofusidate, sodium glycodihydrofusidate, sodium glycolate, sodium lauryl sulfate, dioctyl sodium sulfosuccinate, sucrose fatty acid esters, lactose fatty acid esters, polysorbate, polyoxyethylene-8 lauryl ether, Tween 80, sucrose laurate, macrogol-8 glyceride, lauroyl carbohydrate a gastrointestinal permeation enhancer selected from the group consisting of nitine chloride, caprylic acid, sodium caprylate, sodium caprate (sodium decanoate, C10), sodium cholate, choline geranate, 5-CNAC (N-(5-chlorosalicyloyl)-8-aminocaprylic acid), sodium zalcaprozate (SNAC), chitosan, chitosan glutamate, N-sulfant-N,O-carboxymethylchitosan (SNOCC), N-trimethylated chitosan chloride (TMC), ethanol, penetratin, Zonula occludens toxin (Zot), and polycarbophil-cysteine ​​complex (PCP-Cys); Including, The dosage form does not have a coating that provides substantial protection from the gastric environment in addition to the polypeptide therapeutic agent, alginate oligomer, and gastrointestinal permeation enhancer.

[0401] In other words, the present invention further provides a dosage form adapted for delivery to the stomach of a human or non-human animal subject, said dosage form comprising: (i) a polypeptide therapeutic; and (ii) an alginate oligomer; (iii) EDTA, citric acid, salicylate, oleic acid, lauric acid, acylcarnitine, acylcholine, and acylated amino acids, sodium glycocholate, sodium deoxycholate, sodium taurocholate, sodium dihydrofusidate, sodium glycodihydrofusidate, sodium glycolate, sodium lauryl sulfate, dioctyl sodium sulfosuccinate, sucrose fatty acid esters, lactose fatty acid esters, polysorbate, polyoxyethylene-8 lauryl ether, Tween 80, sucrose laurate, macrogol-8 glyceride, lauroylcarnitine chloroform a gastrointestinal permeation enhancer selected from the group consisting of methylpropional, caprylic acid, sodium caprylate, sodium caprate (sodium decanoate, C10), sodium cholate, choline geranate, 5-CNAC (N-(5-chlorocylic acidoyl)-8-aminocaprylic acid, sodium zalcaprozate (SNAC), chitosan, chitosan glutamate, N-sulfant-N,O-carboxymethylchitosan (SNOCC), N-trimethylated chitosan chloride (TMC), ethanol, penetratin, Zonula occludens toxin (Zot), and polycarbophil-cysteine ​​complex (PCP-Cys), and, optionally, (iv) a pharmaceutically acceptable excipient, said excipient being not an additional compound that provides substantial protection from the stomach environment; consists of, and The dosage form does not have a coating that provides substantial protection from the gastric environment in addition to the polypeptide therapeutic agent, alginate oligomer and gastrointestinal permeation enhancer, and excipients, if used.

[0402] In other embodiments, any of the gastrointestinal permeation enhancers disclosed herein, such as sodium N-(8-[2-hydroxybenzoyl]amino)caprylate (SNAC), decanoic acid and its salts (C10), octanoic acid and its salts (C8), dodecanoic acid and its salts (C12), icosanoic acid and its salts (C20), palmitoylcarnitine chloride, lauroylcarnitine, sodium salicylate, 3-methoxysalicylic acid, 5-methoxysalicylic acid, homovanillic acid, sodium lauroylalanine, N-dodecanoyl-L-glutamic acid, sodium caproylsarcosine, sodium stearoylglutamine (e.g., Amisoft 20 HS-11), and the like, may be used. P), C8-10 alkyl polysaccharides (e.g., Multitrope™ 1620-LQ-(MV)), N-dodecyl-β-D-maltoside, ethylene glycol tetraacetic acid (EGTA), polyoxyl-35 castor oil (also known as Cremophor EL and Koriphor EL (CAS number 61791-12-6));Trimethylchitosan, N,N,N-trimethylchitosan, benzalkonium chloride, cetylpyridinium chloride, cetyltrimethylammonium bromide, ethanol, oleyl alcohol, diethylene glycol monoethyl ether (Transcutol), 1-dodecylazacycloheptan-2-one (Azone®), Labrasol® or Aconon® MC8-2, phosphatidylcholine, sodium cholate, dodecylphosphocholine, rhamnolipid carbopol 934P, octylpyrrolidone, sodium taurocholate, 8-(N-2 -Hydroxy-5-chloro-benzoyl)amino-caprylic acid (5-CNAC), N-(10-[2-hydroxybenzoyl]amino)decanoic acid (SNAD), D-α-tocopheryl polyethylene glycol-1000 succinate (TPGS), sodium decanoate (capric acid), octyl gallate, sodium octanoate (sodium caprylate), lauryl gallate, dodecyl maltoside (DDM), tetradecyl maltoside (TDM), palmitoyl carnitine, sodium chenodeoxycholate (NaCDC), polyoxyl 15-hydroxystearate (Colifor HS 15), sodium deoxycholate, polyoxyethylene (10) oleyl ether (Bridge 10), sodium tetraglycocholate, polyethylene glycol hexadecyl ether, sodium ursodeoxycholate, polyoxyethylene (10) cetyl ether, sodium glycocholate, Pure Bright PEG, sodium taurocholate, diethylene glycol monoethyl ether (Transcutol ES), chenodeoxycholic acid, lauroglycol FCC, ethyl gallate, propyl gallate (PG), chitosan, and arachidonic acid may be present in section (iii). SNAC and C10 (especially the sodium salt form, sodium caprate), alone or in combination, are important enhancers that may be present in section (iii).

[0403] The methods, uses, and compositions of the present invention may include several / multiple gastrointestinal permeation enhancers. Thus, the methods, uses, and compositions of the present invention may include at least one, or one or more gastrointestinal permeation enhancers (e.g., at least two, three, four, or five gastrointestinal permeation enhancers; or two, three, four, five, or more gastrointestinal permeation enhancers). These may be structural or functional analogs of each other, or gastrointestinal permeation enhancers from different structural or functional classes. References herein to "a" or "the" gastrointestinal permeation enhancer should be interpreted as covering such embodiments, unless otherwise indicated. In particular, compositions described herein as "consisting of" a set of components of which a "gastrointestinal permeation enhancer" forms a part include a combination of gastrointestinal permeation enhancers as said components.

[0404] The detailed discussion above regarding dosage forms of the present invention applies mutatis mutandis to these aspects of the invention.

[0405] In certain embodiments of the above, the alginate oligomer is a high G, e.g., at least 80% or 85% G, alginate oligomer. In certain embodiments of the above, the alginate oligomer has 2-40 monomer residues and a weight average molecular weight of, e.g., 2600 Da. In certain embodiments of the above, the gastrointestinal permeation enhancer is SNAC and / or sodium caprate. In more specific embodiments of the above, all of these features apply.

[0406] In a more specific embodiment of the above, the alginic acid oligomer has 5-20 monomer residues and a weight average molecular weight of, for example, 3200 Da. In a more specific embodiment of the above, the alginic acid oligomer has 90-95% G residues. In a more specific embodiment of the above, the gastrointestinal permeation enhancer is SNAC and / or sodium caprate. In a more specific embodiment of the above, all of these features apply.

[0407] As shown in the Examples, it was surprisingly found that the systemic bioavailability (gastric absorption) of tirzepatide and retatortide (LY3437943) can be significantly increased when these compounds are orally administered with C10 (sodium caprate). A similar combination of semaglutide and SNAC when administered orally had only a slight effect on bioavailability at 3 and 8 hours, and the actual inconspicuous uptake that occurred was slow. In contrast, tirzepatide and C10, and retatortide and C10, showed rapid and significant absorption in rats.

[0408] Thus, in a further aspect, the present invention provides a method for increasing the systemic bioavailability of tirzepatide, retatortide, or a structural analog thereof when administered orally, by oral gavage, by nasogastric gavage, or intragastric administration, said method comprising administering tirzepatide, and / or retatortide, and / or a structural analog thereof together with capric acid or a pharmaceutically acceptable salt thereof to the stomach of a human or non-human subject as part of one or more dosage forms, wherein said one or more dosage forms, in addition to tirzepatide, retatortide, or a structural analog thereof and capric acid or a pharmaceutically acceptable salt thereof, do not have a coating that provides substantial protection from the gastric environment.

[0409] The present invention further provides capric acid or a pharmaceutically acceptable salt thereof for use in a method for increasing the systemic bioavailability of tirzepatide, retatortide, or structural analogs thereof when administered orally, by oral gavage, nasogastric gavage, or intragastric administration, said method comprising administering tirzepatide, and / or retatortide, and / or structural analogs thereof together with said capric acid or pharmaceutically acceptable salts thereof to the stomach of a human or non-human animal as part of one or more dosage forms, wherein said one or more dosage forms, in addition to tirzepatide, retatortide, or structural analogs and capric acid or pharmaceutically acceptable salts thereof, do not have a coating that provides substantial protection from the gastric environment.

[0410] Similar to other aspects of the invention, this aspect of the invention provides: (i) a method for gastric uptake of [active or reversibly reduced in activity] tirzepatide, retatortide, or structural analogs thereof; (ii) a method for absorption of [active or reversibly reduced in activity] tirzepatide, retatortide, or structural analogs thereof from the stomach of a human subject or a non-human animal subject; (iii) a method for increasing absorption of [active or reversibly reduced in activity] tirzepatide, retatortide, or structural analogs thereof from the stomach of a human subject or a non-human animal subject; (iv) a disease or condition that is responsive to tirzepatide, retatortide, or structural analogs thereof or that is prevented by a polypeptide therapeutic agent, or a disease or condition thereof. and (v) a method for the systemic treatment or prevention of a disease or condition responsive to or prevented by tirzepatide, retatortide, or a structural analog thereof, or a complication thereof, the relevant detailed discussion of which applies mutatis mutandis to these aspects, to the extent that it relates to tirzepatide, retatortide, or a structural analog thereof and / or capric acid or a pharmaceutically acceptable salt thereof.

[0411] The present invention further provides a dosage form adapted for delivery to the stomach of a human or non-human animal subject, said dosage form comprising: (i) tirzepatide, and / or retatortide, and / or structural analogs thereof; (ii) capric acid or a pharmaceutically acceptable salt thereof; Including, The dosage form does not have a coating that provides substantial protection from the gastric environment in addition to tirzepatide, and / or retatortide, and / or structural analogs thereof, and capric acid or a pharmaceutically acceptable salt thereof.

[0412] In other words, the present invention further provides a dosage form adapted for delivery to the stomach of a human or non-human subject, said dosage form comprising: (i) tirzepatide, and / or retatortide, and / or structural analogs thereof; (ii) capric acid or a pharmaceutically acceptable salt thereof, and optionally (iii) a pharmaceutically acceptable excipient, said excipient being not an additional compound that provides substantial protection from the stomach environment; consists of, and The dosage form contains tirzepatide, retatortide or a structural analog thereof, capric acid or a pharmaceutically acceptable salt thereof, and excipients, if used, and does not have a coating that provides substantial protection from the gastric environment.

[0413] As discussed above, tirzepatide and retatortide are considered structural analogs of each other, and therefore component (i) of these aspects of the invention can be any of the compounds described above in connection with tirzepatide and retatortide (LY3437943).

[0414] Pharmaceutically acceptable salts of capric acid may be any of the base addition salts disclosed above in connection with therapeutic polypeptides, in particular alkali metal salts such as sodium or potassium salts, alkaline earth metal salts such as calcium or magnesium salts, zinc salts or ammonium salts.

[0415] The above detailed discussion of the dosage forms of the present invention applies mutatis mutandis to these aspects of the invention as they relate to tirzepatide, retatortide, or structural analogs thereof and / or capric acid or a pharmaceutically acceptable salt thereof.

[0416] The subject may be any human or non-human animal subject, particularly a human or non-human vertebrate, such as a non-human mammal, bird, amphibian, fish, or reptile. In a preferred embodiment, the subject is a mammalian subject. The animal may be livestock or commercially valuable livestock or animals, including laboratory animals or animals in zoos or game reserves. Thus, representative animals include dogs, cats, rabbits, mice, guinea pigs, hamsters, horses, pigs, sheep, goats, and cows. Thus, veterinary uses of the present invention are encompassed. The subject may be referred to as a patient. Preferably, the subject is a human. In some embodiments, the subject is not a ruminant mammal.

[0417] The vertebrate gastrointestinal (GI) tract, also known as the digestive tract or alimentary canal, is a continuous system of organs beginning at the mouth and ending at the anus. Specifically, this system consists of the mouth, pharynx, esophagus, stomach (or stomach in ruminant mammals), duodenum, small intestine, large intestine, and anus. For purposes of the present invention, these organs can be subdivided into an upper GI tract consisting of the mouth, pharynx, esophagus, and stomach, and a lower GI tract (intestinal tract) consisting of the duodenum, jejunum, ileum (together with the small intestine), cecum, colon, rectum (together with the large intestine), and anus. When the present invention applies to ruminants, references to the "stomach" should be considered references to the "abomasum."

[0418] "Treatment," as generally used in reference to the treatment of a disease or medical condition in a subject according to the present invention, is used broadly herein to include any therapeutic effect, i.e., any beneficial effect on the disease or condition. Accordingly, it includes not only eradication or elimination of the disease or condition, or curing the subject, but also improvement in the subject's disease or condition. Thus, it includes, for example, improvement in any signs or symptoms of the disease or condition, or in any clinically recognized indicator of the disease / condition. Thus, treatment includes both curative and palliative therapy, i.e., reactive treatment, of, for example, a known or diagnosed disease / condition.

[0419] "Prevention," as generally used herein, refers to any preventative or preventative effect. Thus, it includes delaying, limiting, reducing, or preventing a disease, symptom, or the onset of a disease or symptom, or one or more signs or indicators thereof, for example, compared to the disease, symptom, or sign or indicator before the prophylactic treatment. Thus, prophylaxis explicitly includes both absolute prevention of the occurrence or development of a disease or symptom, or its signs or indicators, and any delay in the onset or development of a disease, symptom, sign, or indicator, or reduction or limitation of the development or progression of a disease, symptom, sign, or indicator.

[0420] Those skilled in the art will be able to formulate the alginate oligomers and therapeutic polypeptides and gastrointestinal permeation enhancers for use in the present invention, alone or in various combinations, into pharmaceutical compositions and dosage forms suitable for use in the above-described aspects and embodiments of the present invention by any of the conventional methods known in the art and widely described in the literature.

[0421] More particularly, the alginate oligomers for use in the present invention may be incorporated together with a polypeptide therapeutic agent and / or a gastrointestinal permeation agent, if desired together with one or more conventional carriers, diluents and / or excipients, to produce conventional galenical formulations such as tablets, pills, granules, powders, troches, sachets, cachets, elixirs, suspensions, emulsions, solutions, syrups, soft and hard gelatin capsules and the like.

[0422] Similarly, the polypeptide therapeutic agents for use in the present invention may be incorporated with one or more conventional carriers, diluents and / or excipients, and, if desired, alginate oligomers and / or gastrointestinal permeation agents, to produce conventional galenic formulations such as tablets, pills, granules, powders, troches, sachets, cachets, elixirs, suspensions, emulsions, solutions, syrups, soft and hard gelatin capsules and the like.

[0423] Similarly, the gastrointestinal permeation agents for use in the present invention may be incorporated together with the alginate oligomer and / or polypeptide therapeutic agent and one or more conventional carriers, diluents and / or excipients, as appropriate, to produce conventional galenical formulations such as tablets, pills, granules, powders, troches, sachets, cachets, elixirs, suspensions, emulsions, solutions, syrups, soft and hard gelatin capsules and the like.

[0424] Such formulations may be, for example, any pharmaceutically acceptable composition in the form of a solution, dispersion, emulsion, powder, tablet, capsule, gel, and the like, containing any of the formulation ingredients specifically named herein. Conveniently, the formulation will be in the form of a tablet or liquid or powder / pellet filled capsule, although this is not an exhaustive list.

[0425] Examples of suitable carriers, excipients and diluents are lactose, dextrose, sucrose, sorbitol, mannitol, starch, gum arabic, calcium phosphate, tragacanth, gelatin, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, water syrup, water, water / ethanol, water / glycol, water / polyethylene, hypertonic saline, glycol, propylene glycol, methylcellulose, methyl hydroxybenzoates, propyl hydroxybenzoate, talc, magnesium stearate, mineral oil or fatty substances such as hard fats, or suitable mixtures thereof. Important excipients and diluents are mannitol and hypertonic saline (saline).

[0426] The compositions may further comprise lubricating agents, wetting agents, emulsifying agents, suspending agents, preservatives, sweetening agents, flavoring agents, buffers, and the like. In certain embodiments, the polypeptide therapeutic agent, alginate oligomer, and, if used, gastrointestinal permeation enhancers, and more generally, the dosage form does not include agents that provide substantial protection from the gastric environment, such as buffers and enzyme (protease) inhibitors.

[0427] Excipients for tablets, capsules, and solid or semi-solid mixed formulations may include diluents, binders, lubricants, disintegrants, flow agents, stabilizers, and surfactants. Sweeteners, flavoring agents, and coloring agents may also be added to obtain an acceptable product, even if they do not directly affect the performance of the formulation.

[0428] In solid dosage forms, cellulose, starch, monosaccharides, disaccharides, carbonates and polyvinylpyrrolidone may act as fillers, binders and disintegrants. Fatty acids, stearates and silicates may act as lubricants in solid dosage forms.

[0429] In addition to the water (e.g., purified or sterilized) vehicle, excipients for aqueous liquid formulations may include cosolvents, buffers, surfactants, rheology modifiers, preservatives, and antioxidants. Sweeteners, flavoring agents, and coloring agents may also be added to obtain an acceptable product, even if they do not directly affect the performance of the formulation.

[0430] Compositions or dosage forms comprising the subject polypeptide therapeutic agents, particularly the lipid-modified incretin analogs described herein, may be prepared using a variety of surfactants, including, for example, poly(ethylene glycol), including poly(ethylene glycol) having a molecular weight ranging from about 200 to about 5,000 Da, ethylene glycol, propylene glycol, non-ionic surfactants, tyloxapol, polysorbate 20, polysorbate 80, macrogol-15-hydroxystearate, phospholipids, lecithin, dimyristoylphosphatidylcholine, dipalmitoylphosphatidylcholine, distearoylphosphatidylcholine, cyclodextrins, α-cyclodextrin, β-cyclodextrin, γ-cyclodextrin, hydroxyethyl-β-cyclodextrin, hydroxypropyl-β-cyclodextrin, hydroxyethyl-γ-cyclodextrin, hydroxypropyl-γ-cyclodextrin, dihydroxypropyl-β ... The composition may include one or more solubility enhancers such as cyclodextrin, sulfobutylether-β-cyclodextrin, sulfobutylether-γ-cyclodextrin, glucosyl-α-cyclodextrin, glucosyl-β-cyclodextrin, diglucosyl-β-cyclodextrin, maltosyl-α-cyclodextrin, maltosyl-β-cyclodextrin, maltosyl-γ-cyclodextrin, maltotriosyl-β-cyclodextrin, maltotriosyl-γ-cyclodextrin, dimaltosyl-β-cyclodextrin, methyl-β-cyclodextrin, carboxyalkyl thioether, hydroxypropyl methylcellulose, hydroxypropyl cellulose, polyvinylpyrrolidone, vinyl acetate copolymer, vinylpyrrolidone, sodium lauryl sulfate, dioctyl sodium sulfosuccinate, or any combination thereof.

[0431] The alginate oligomer may be administered in a dose of 0.01 to 10 g, for example 0.05, 0.1, 0.2, 0.4, 0.6, 0.8, 1.0, 1.5, 2.0, 2.5, 3.0, 3.5, 4.0, 5.0, 6.0, 7.0, 8.0, 9.0 to 10 g, or 0.05 to 0.1, 0.2, 0.4, 0.6, 0.8, 1.0, 1.5, 2.0, 2.5, 3.0, 3.5, 4.0, 5.0, 6.0, 7.0, 8.0, 9.0 or 10 g, or 0.1 g to 10 g, 0.5 g to 5 g, 0.8 g to 3 g, or 1 g to 2 g, for example about 2 g.

[0432] A typical tablet used to administer alginate oligomer to the stomach in accordance with the present invention may contain up to 99%, up to 95%, 90%, 85% or 80%, e.g., 50-95%, 55-95%, 60-95%, 65-95%, 70-95%, 75-95%, 80-95%, 85-95%, 90-95%, 50-90%, 50-90%, 55-90%, 60-90%, 65-90%, 70-90%, 75-90%, 80-90%, 85-90%, 50-90%, 55-85%, 60-80%, or 65-75% w / w oligomer, with the remainder consisting of pharmaceutically acceptable excipients and / or other active agents, if used (e.g., therapeutic polypeptides and / or gastrointestinal permeation enhancers).

[0433] A typical powder for oral administration of alginate oligomers may have a saturation of up to 100%, e.g., 99%, 95%, 90%, 85%, 80%, 75% or 70%, e.g., 50 to 90%, 55 to 90%, 60 to 90%, 65 to 90%, 70 to 90%, 75 to 90%, 80 to 90%, 85 to 90%, 50 to 85%, 55 to 85%, 60 to 85%, 65 to 85%, 70 to 85%, 75 to 85%. , 80-85%, 50-80%, 55-80%, 60-80%, 65-80%, 70-80%, 75-80%, 50-70%, 55-70%, 60-70%, or 65-70% w / w alginate oligomer, with the remainder consisting of pharmaceutically acceptable excipients and / or other active agents (e.g., therapeutic polypeptides and / or gastrointestinal permeation enhancers) if used in the same composition.

[0434] Typical solutions used to administer alginate oligomers to the stomach in accordance with the present invention may contain up to 1 to 25%, 1 to 20%, e.g., 1 to 15%, 1 to 10%, 1 to 9%, 1 to 8%, 1 to 7% or 1 to 6%, 5 to 25%, 5 to 20%, 5 to 15%, 5 to 10%, 5 to 9%, 5 to 8%, 5 to 7%, 5 to 6%, 8 to 25%, 8 to 20%, 8 to 15%, 8 to 10%, 9 to 25%, 9 to 20%, or 9 to 15% weight / volume or weight / weight alginate oligomer, with the remainder consisting of pharmaceutically acceptable excipients, e.g., water and / or other active agents, if used (e.g., therapeutic polypeptides and / or gastrointestinal permeation enhancers).

[0435] A typical tablet used to administer a gastrointestinal permeation enhancer to the stomach in accordance with the present invention may contain up to 99%, up to 95%, 90%, 85%, or 80%, e.g., 50-95%, 55-95%, 60-95%, 65-95%, 70-95%, 75-95%, 80-95%, 85-95%, 90-95%, 50-90%, 50-90%, 55-90%, 60-90%, 65-90%, 70-90%, 75-90%, 80-90%, 85-90%, 50-90%, 55-85%, 60-80%, or 65-75% w / w gastrointestinal permeation enhancer, with the remainder consisting of pharmaceutically acceptable excipients and / or other active agents, if used (e.g., therapeutic polypeptide and / or alginate oligomer).

[0436] Representative powders for oral administration of gastrointestinal permeation enhancers after mixing with food or beverages may have a saturation of up to 100%, e.g., up to 99%, 95%, 90%, 85%, 80%, 75%, or 70%, e.g., 50 to 90%, 55 to 90%, 60 to 90%, 65 to 90%, 70 to 90%, 75 to 90%, 80 to 90%, 85 to 90%, 50 to 85%, 55 to 85%, 60 to 85%, 65 to 85%, 70 to 85%, 75 to 85% , 80-85%, 50-80%, 55-80%, 60-80%, 65-80%, 70-80%, 75-80%, 50-70%, 55-70%, 60-70%, or 65-70% w / w gastrointestinal permeation enhancer, with the remainder consisting of pharmaceutically acceptable excipients and / or other active agents (e.g., therapeutic polypeptides and / or alginate oligomers) if used in the same composition.

[0437] Typical solutions used to administer a gastrointestinal permeation enhancer to the stomach in accordance with the present invention may contain up to 1 to 25%, 1 to 20%, e.g., 1 to 15%, 1 to 10%, 1 to 9%, 1 to 8%, 1 to 7%, or 1 to 6%, 5 to 25%, 5 to 20%, 5 to 15%, 5 to 10%, 5 to 9%, 5 to 8%, 5 to 7%, 5 to 6%, 8 to 25%, 8 to 20%, 8 to 15%, 8 to 10%, 9 to 25%, 9 to 20%, or 9 to 15% weight / volume or weight / weight of gastrointestinal permeation enhancer, with the remainder consisting of pharmaceutically acceptable excipients, such as water and / or other active agents (e.g., therapeutic polypeptides and / or alginate oligomers, if used).

[0438] Exemplary tablets used to administer polypeptide therapeutic agents to the stomach in accordance with the present invention may contain up to 10%, up to 9.5%, 9.0%, 8.5%, 8.0%, 7.5%, 7.0%, 6.5%, 6.0%, 5.5%, 5.0%, 4.5%, 4.0%, 3.5%, 3.0%, 2.5%, 2.0%, 1.5%, 1.0%, 0.5%, or 0.1%, e.g., 0.1, 0.2, 0.5, 1.0, 1.5, 2.0, 2.5, 3.0, 3.5, 4.0, 4.5, 5.0, 5.5, 6.0, 6.5, 7.0, 7.5, 8.0, 8.5, 9.0, 9.5 to 10%, or 0.1 to 0.2, 0.5, 1.0, 1.5, 2.0, 2.5, 3.0, 3.5, 4.0, 4.5, 5.0, 5.5, 6.0, 6.5, 7.0, 7.5, 8.0, 8.5, 9.0, 9.5, or or 10%, or 5.0 to 9.5%, 5.5 to 9.5%, 6.0 to 9.5%, 6.5 to 9.5%, 7.0 to 9.5%, 7.5 to 9.5%, 8.0 to 9.5%, 8.5 to 9.5%, 9.0 to 9.5%, 5.0 to 9.0%, 5.0 to 9.0%, 5.5 to 9.0%, 6.0 to 9.0%, 6.5 to 9.0%, 7.0 to 9.0%, 7.5 to 9. The composition may contain 0%, 8.0 to 9.0%, 8.5 to 9.0%, 5.0 to 9.0%, 5.5 to 8.5%, 6.0 to 8.0%, or 6.5 to 7.5% weight / weight of polypeptide therapeutic agent, with the remainder consisting of pharmaceutically acceptable excipients and / or other active agents, if used (e.g., gastrointestinal permeation enhancers and / or alginate oligomers).

[0439] Representative powders for oral administration of polypeptide therapeutics after mixing with food or beverages may contain up to 10%, up to 9.5%, 9.0%, 8.5%, 8.0%, 7.5%, 7.0%, 6.5%, 6.0%, 5.5%, 5.0%, 4.5%, 4.0%, 3.5%, 3.0%, 2.5%, 2.0%, 1.5%, 1.0%, 0.5%, or 0.1%, e.g., 0.1, 0.2, 0.5, 1.0%. 0.0, 1.5, 2.0, 2.5, 3.0, 3.5, 4.0, 4.5, 5.0, 5.5, 6.0, 6.5, 7.0, 7.5, 8.0, 8.5, 9.0, 9.5 to 10%, or 0.1 to 0.2, 0.5, 1.0, 1.5, 2.0, 2.5, 3.0, 3.5, 4.0, 4.5, 5.0, 5.5, 6.0, 6.5, 7.0, 7.5, 8.0, 8.5, 9.0, 9.5, or 1 0%, or 5.0 to 9.5%, 5.5 to 9.5%, 6.0 to 9.5%, 6.5 to 9.5%, 7.0 to 9.5%, 7.5 to 9.5%, 8.0 to 9.5%, 8.5 to 9.5%, 9.0 to 9.5%, 5.0 to 9.0%, 5.0 to 9.0%, 5.5 to 9.0%, 6.0 to 9.0%, 6.5 to 9.0%, 7.0 to 9.0%, 7.5 to 9.0%, 8. It may contain 0 to 9.0%, 8.5 to 9.0%, 5.0 to 9.0%, 5.5 to 8.5%, 6.0 to 8.0%, or 6.5 to 7.5% weight / weight of polypeptide therapeutic agent, with the remainder consisting of pharmaceutically acceptable excipients and / or other active agents (e.g., gastrointestinal permeation enhancers and / or alginate oligomers) if used in the same composition.

[0440] Exemplary solutions used to administer polypeptide therapeutic agents to the stomach according to the present invention may contain up to 10%, up to 9.5%, 9.0%, 8.5%, 8.0%, 7.5%, 7.0%, 6.5%, 6.0%, 5.5%, 5.0%, 4.5%, 4.0%, 3.5%, 3.0%, 2.5%, 2.0%, 1.5%, 1.0%, 0.5%, or 0.1%, e.g., 0.1, 0.2, 0.5, 1.0%, , 1.5, 2.0, 2.5, 3.0, 3.5, 4.0, 4.5, 5.0, 5.5, 6.0, 6.5, 7.0, 7.5, 8.0, 8.5, 9.0, 9.5 to 10%, or 0.1 to 0.2, 0.5, 1.0, 1.5, 2.0, 2.5, 3.0, 3.5, 4.0, 4.5, 5.0, 5.5, 6.0, 6.5, 7.0, 7.5, 8.0, 8.5, 9.0, 9.5, or 10%, or or 5.0 to 9.5%, 5.5 to 9.5%, 6.0 to 9.5%, 6.5 to 9.5%, 7.0 to 95%, 7.5 to 9.5%, 8.0 to 9.5%, 8.5 to 9.5%, 9.0 to 9.5%, 5.0 to 9.0%, 5.0 to 9.0%, 5.5 to 9.0%, 6.0 to 9.0%, 6.5 to 9.0%, 7.0 to 9.0%, 7.5 to 9.0%, 8.0 to 9.0 %, 8.5 to 9.0%, 5.0 to 9.0%, 5.5 to 8.5%, 6.0 to 8.0%, or 6.5 to 7.5% weight / volume or weight / weight of polypeptide therapeutic agent, with the remainder consisting of pharmaceutically acceptable excipients, e.g., water, and / or other active agents, if used (e.g., gastrointestinal permeation enhancers and / or alginate oligomers).

[0441] A typical tablet used to administer an alginate oligomer, a gastrointestinal permeation enhancer, and a polypeptide therapeutic agent to the stomach in accordance with the present invention may contain 20 to 60% (e.g., 25, 30, 35, 40, 50, or 55 to 60%, or 25 to 30, 35, 40, 50, 55, or 60%) wt / wt alginate oligomer, 20 to 60% (e.g., 25, 30, 35, 40, 50, or 55 to 60%, or 25 to 30, 35, 40, 50, 55, or 60%) wt / volume or wt / wt gastrointestinal permeation enhancer, 1 to 10% (e.g., 2, 3, 4, 5, 6, 7, 8, 9 to 10%, or 2 to 3, 4, 5, 6, 7, 8, 9, or 10%) wt / volume or wt / wt polypeptide therapeutic agent, up to a maximum of 100%, with the remainder consisting of pharmaceutically acceptable excipients.

[0442] A typical powder for oral administration of an alginate oligomer, a gastrointestinal permeation enhancer, and a polypeptide therapeutic agent after admixture with a food or beverage may contain 20 to 60% (e.g., 25, 30, 35, 40, 50, or 55 to 60% or 25 to 30, 35, 40, 50, 55, or 60%) wt / vol or wt / wt alginate oligomer, 20 to 60% (e.g., 25, 30, 35, 40, 50, or 55 to 60% or 25 to 30, 35, 40, 50, 55, or 60%) wt / vol or wt / wt gastrointestinal permeation enhancer, 1 to 10% (e.g., 2, 3, 4, 5, 6, 7, 8, 9 to 10% or 2 to 3, 4, 5, 6, 7, 8, 9, or 10%) wt / wt polypeptide therapeutic agent, up to a maximum of 100%, with the remainder consisting of pharmaceutically acceptable excipients.

[0443] A typical solution used to administer an alginate oligomer, a gastrointestinal permeation enhancer, and a polypeptide therapeutic agent to the stomach in accordance with the present invention may comprise 20 to 60% (e.g., 25, 30, 35, 40, 50, or 55 to 60%) weight / volume or weight / weight alginate oligomer, 20 to 60% (e.g., 25, 30, 35, 40, 50, or 55 to 60%) weight / volume or weight / weight alginate oligomer, It may contain 5 to 60% or 25 to 30, 35, 40, 50, 55, or 60%) weight / volume or weight / weight of a gastrointestinal permeation enhancer, 1 to 10% (e.g., 2, 3, 4, 5, 6, 7, 8, 9 to 10%, or 2 to 3, 4, 5, 6, 7, 8, 9, or 10%) weight / volume or weight / weight of a polypeptide therapeutic agent, up to a maximum of 100%, with the remainder consisting of pharmaceutically acceptable excipients.

[0444] Tablets for use in the present invention may comprise a total of about 0.1% to 100% weight / weight of polypeptide therapeutic agent and alginate oligomer, and a total of 0 to about 99.9% weight / weight of gastrointestinal permeation enhancer and / or additional excipients. For example, the tablet may contain, in total, about 0.5, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, or 95% to 100%, or about 0.1% to about 0.5, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, or 95% polypeptide therapeutic agent and alginate oligomer, in total. and about 0.1, 0.5, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, or 95% to about 99%, or about 0.1% to about 0.5, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, or 99% of the gastrointestinal permeation enhancer and / or additional excipient. Any ranges with endpoints formed from any of these values ​​are expressly contemplated.

[0445] In these embodiments, the weight ratio of polypeptide therapeutic agent to alginate oligomer in the tablet is from about 1:0.5 to about 1:100, e.g., about 1:1, 1:2, 1:3, 1:4, 1:5, 1:6, 1:7, 1:8, 1:9, 1:10, 1:15, 1:20, 1:25, 1:30, 1:35, 1:40, 1:45, 1:50, 1:55, 1:60, 1:65, 1:70, 1:75, 1:80, 1:95, 1:100, 1:110, 1:120, 1:130, 1:140, 1:150, 1:160, 1:170, 1:180, 1:190, 1:200, 1:210, 1:220, 1:230, 1:240, 1:250, 1:260, 1:270, 1:280, 1:290, 1:300, 1:310, 1:320, 1:330, 1:340, 1:350, 1:360, 1:370, 1:380, 1:390, 1:410, 1:420, 1:430, 1:440, 1:450, 1:510, 1:520, 1:530, 1:540, 1:550, 1:560, 1:570, 1:580, 1:590, 1:610, 1:620, 1:6 It can be from 0, 1:85, 1:90, or 1:95 to about 1:100, or from about 1:0.5 to about 1:1, 1:2, 1:3, 1:4, 1:5, 1:6, 1:7, 1:8, 1:9, 1:10, 1:15, 1:20, 1:25, 1:30, 1:35, 1:40, 1:45, 1:50, 1:55, 1:60, 1:65, 1:70, 1:75, 1:80, 1:85, 1:90, or 1:95. Any ranges with endpoints formed from any of these values ​​are expressly contemplated.

[0446] In these embodiments, the weight ratio of gastrointestinal permeation enhancer to the total amount of polypeptide therapeutic agent and alginate oligomer in the tablet is from about 1:0.5 to about 1:100, e.g., about 1:1, 1:2, 1:3, 1:4, 1:5, 1:6, 1:7, 1:8, 1:9, 1:10, 1:15, 1:20, 1:25, 1:30, 1:35, 1:40, 1:45, 1:50, 1:55, 1:60, 1:65, 1:70, 1:80, 1:90, 1:100, 1:110, 1:120, 1:130, 1:140, 1:150, 1:160, 1:170, 1:180, 1:190, 1:200, 1:210, 1:220, 1:230, 1:240, 1:250, 1:260, 1:270, 1:280, 1:290, 1:300, 1:310, 1:320, 1:330, 1:340, 1:350, 1:360, 1:370, 1:380, 1:390, 1:410, 1:420, 1:430, 1:440, 1:450, 1:460, 1:470, 1:480, 1:490, 1:510, 1:520, 1:530, 1:540, 1:550, 1:560, 1 The range may be from about 1:0.5 to about 1:1, 1:2, 1:3, 1:4, 1:5, 1:6, 1:7, 1:8, 1:9, 1:10, 1:15, 1:20, 1:25, 1:30, 1:35, 1:40, 1:45, 1:50, 1:55, 1:60, 1:65, 1:70, 1:75, 1:80, 1:85, 1:90, or 1:95 to 1:100, or from about 1:0.5 to about 1:1, 1:2, 1:3, 1:4, 1:5, 1:6, 1:7, 1:8, 1:9, 1:10, 1:15, 1:20, 1:25, 1:30, 1:35, 1:40, 1:45, 1:50, 1:55, 1:60, 1:65, 1:70, 1:75, 1:80, 1:85, 1:90, or 1:95. Any ranges with endpoints formed from any of these values ​​are expressly contemplated.

[0447] In these embodiments, the tablets contain from about 0.1 to about 200 mg, for example, from about 0.5, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 105, 110, 115, 120, 125, 130, 135, 140, 145, 150, 155, 160, 165, 170, 175, 180, 185, 190, 195 to about 200 mg, or from about 0.1 to about 0.5, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 105, 110, 115, 120, 125, 130, 135, 140, 145, 150, 155, 160, 165, 170, 175, 180, 185, 190, 195 to about 200 mg. 7, 8, 9, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 105, 110, 115, 120, 125, 130, 135, 140, 145, 150, 155, 160, 165, 170, 175, 180, 185, 190 or 195 mg of a polypeptide therapeutic and about 0.1 to about 2000 mg, e.g., about 0.5, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 20, 30, 40, 50, 60, 70, 80, 90, 100 , 150, 200, 250, 300, 350, 400, 450, 500, 550, 600, 650, 700, 750, 800, 850, 900, 950, 1000, 1050, 1100, 1150, 1200, 1250, 1300, 1350, 1400, 1450, 1500, 1550, 1600, 1650, 1700, 1750, 1800, 1850, 1900, 1950 to about 2000 mg, or about 0.1 to about 0.5, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 20, 30, 40, 50, 6 0, 70, 80, 90, 100, 150, 200, 250, 300, 350, 400, 450, 500, 550, 600, 650, 700, 750, 800, 850, 900, 950, 1000, 1050, 1100, 1150, 1200, 1250, 1300, 1350, 1400, 1450, 1500, 1550, 1600, 1650, 1700, 1750, 1800, 1850, 1900, or 1950 mg of alginate oligomer, totaling 0 to about 2000 mg, e.g., about 0.1, 0.5, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 20, 30, 40, 50, 60, 70, 80, 90, 100, 150, 200, 250, 300, 350, 400, 450, 500, 550, 600, 650, 700, 750, 800, 850, 900, 950, 1000, 1050, 1100, 1150, 1200, 1250, 1300, 1350, 1400, 1450, 1500, 1550, 1600, 1650, 1700, 1750, 1800, 1850, 1900, 1950 to about 2000 mg, or 0 to about 0.1, 0.5, 1, 2, 3 , 4, 5, 6, 7, 8, 9, 10, 20, 30, 40, 50, 60, 70, 80, 90, 100, 150, 200, 250, 300, 350, 400, 450, 500, 550, 600, 650, 700, 750, 800, 850, 900, 950, 1000, 1050, 1100, 1150, 1200, 1250, 1300, 1350, 1400, 1450, 1500, 1550, 1600, 1650, 1700, 1750, 1800, 1850, 1900, or 1950 mg of gastrointestinal permeation enhancer and / or additional excipients.

[0448] Film-coated tablets for use in the present invention may comprise a core having the tablet parameters described above and a film coating the core, e.g., about 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 μm thick, comprising active or inactive film-forming ingredients, e.g., lipids, PEG, and colorants. In these embodiments, the coating comprises less than about 50%, e.g., less than 45, 40, 35, 30, 25, 20, 15, 10, or 5%, of the total weight of the formulation.

[0449] Slow- or extended-release tablets (depot tablets) for use in the present invention comprise a total of 0.1% to 100% w / w of polypeptide therapeutic agent and alginate oligomer, and a total of 0 to 99.9% w / w of gastrointestinal permeation enhancer and / or additional excipients, the components of which may be provided in a layered arrangement, such as those described above. The specific parameters for the tablets listed above apply mutatis mutandis to these slow- or extended-release tablets.

[0450] Slow- or extended-release tablets (depot tablets) for use in the present invention may or may not be further provided with an outer film layer, e.g., about 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 μm thick, comprising active or inactive film-forming ingredients, e.g., lipids, PEG, and colorants. In these embodiments, the coating comprises less than about 50%, e.g., less than about 45, 40, 35, 30, 25, 20, 15, 10, or 5%, of the total weight of the formulation.

[0451] Hard capsules for use in the present invention comprise a hard shell formed from gelatin and / or other hard-shell-forming polymeric compounds, e.g., about 10, 20, 30, 40, 50, 60, 70, 80, 90, or 100 μm thick, and may optionally further comprise active or inactive excipients, stabilizers, colorants and flavorings, and a filler having the parameters of an uncoated tablet as described above. The shell may essentially dissolve quickly in the gastric environment. The filler may be particulate, e.g., in the form of powder, granules, pellets, or microparticles or nanoparticles, or a combination thereof. The particulate filler may be monodisperse or polydisperse in size.

[0452] The soft capsules used in the present invention may comprise a formable shell, e.g., about 50, 60, 70, 80, 90, 100, 110, 120, 130, 140, or 150 μm thick, formed from gelatin and / or other easily formable shell-forming polymeric compounds and a suitable solvent that maintains formability, and optionally further comprising an active agent or inactive excipients, stabilizers, colorants, and flavorings, and a liquid or semisolid, e.g., gel, fill material, having the parameters of an uncoated tablet described above. The shell may dissolve essentially immediately in the gastric environment. In these embodiments, the fill material typically comprises a liquid excipient, particularly an aqueous or oil-based liquid excipient. The fill material may be a liquid or gel phase containing other ingredients in particulate form or particles formed from other ingredients, e.g., powders, granules, pellets, or microparticles or nanoparticles, or a combination thereof. The particulate components may be monodisperse or polydisperse in size. Gels may be formed from any gel-forming excipient, particularly hydrogel-forming excipients such as gelatin, cellulose or alginate polymers.

[0453] In other embodiments, the formulations for use in the present invention may be solid or semi-solid, e.g., gelled, mixtures of components having the parameters of the uncoated tablets described above. These mixtures may be in the form of powders, granules, pellets, or microparticles or nanoparticles, or a combination thereof. The particles may be monodisperse or polydisperse in size. Individual components of the formulation, such as the polypeptide therapeutic agent, alginic acid oligomer, gastrointestinal permeation enhancer, or other excipients, may be provided as or on separate particle types, or a combination of some or all of the components may be provided on different particle types. Gels may be formed from any gel-forming excipient, particularly hydrogel-forming excipients, such as gelatin, cellulose, or alginate polymers. The gel may contain the above-described particulate or non-particulate forms of the individual components of the formulation.

[0454] In certain embodiments, these solid or semi-solid mixtures may be designed to be dissolved, dispersed or suspended in a pharmaceutically acceptable liquid, such as water, prior to administration by oral, oral gavage, nasogastric gavage and intragastric routes.

[0455] Other formulations for use in the present invention may contain from about 0.1 to about 200 mg, for example, from about 0.5, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 105, 110, 115, 120, 125, 130, 135, 140, 145, 150, 155, 160, 165, 170, 175, 180, 185, 190, 195 to about 200 mg, or from about 0.1 to about 0.5, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 105, 110, 115, 120, 125, 130, 135, 140, 145, 150, 155, 160, 165, 170, 175, 180, 185, 190, 195 to about 200 mg. , 8, 9, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 105, 110, 115, 120, 125, 130, 135, 140, 145, 150, 155, 160, 165, 170, 175, 180, 185, 190, or 195 mg of a polypeptide therapeutic agent and about 0.1 to about 2000 mg, e.g., about 0.5, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 20, 30, 40, 50, 60, 70, 80, 90, 100, 105, 110, 115, 120, 125, 130, 135, 140, 145, 150, 155, 160, 165, 170, 175, 180, 185, 190, or 195 mg of a polypeptide therapeutic agent. 0, 150, 200, 250, 300, 350, 400, 450, 500, 550, 600, 650, 700, 750, 800, 850, 900, 950, 1000, 1050, 1100, 1150, 1200, 1250, 1300, 1350, 1400, 1450, 1500, 1550, 1600, 1650, 1700, 1750, 1800, 1850, 1900, 1950 to about 2000 mg, or about 0.1 to about 0.5, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 20, 30, 40, 50, 60, 70, 80, 90, 100, 150, 200, 250, 300, 350, 400, 450, 500, 550, 600, 650, 700, 750, 800, 850, 900, 950, 1000, 1050, 1100, 1150, 1200, 1250, 1300, 1350, 1400, 1450, 1500, 1550, 1600, 1650, 1700, 1750, 1800, 1850, 1900, or 1950 mg of alginate oligomer, totaling 0 to about 2000 mg, e.g., about 0.1, 0.5, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 20, 30, 40, 50, 60, 70, 80, 90, 100, 150, 200, 250, 300, 350, 400, 450, 500, 550, 600, 650, 700, 750, 800, 850, 900, 950, 1000, 105 0, 1100, 1150, 1200, 1250, 1300, 1350, 1400, 1450, 1500, 1550, 1600, 1650, 1700, 1750, 1800, 1850, 1900, 1950 to about 2000 mg, or 0 to about 0.1, 0.5, 1, 2, 3, 4, 5, 6 , 7, 8, 9, 10, 20, 30, 40, 50, 60, 70, 80, 90, 100, 150, 200, 250, 300, 350, 400, 450, 500, 550, 600, 650, 700, 750, 800, 850, 900, 950, 1000, 1050, 1100, 1150, 1200, 1250, 1300, 1350, 1400, 1450, 1500, 1550, 1600, 1650, 1700, 1750, 1800, 1850, 1900, or 1950 mg of a gastrointestinal permeation enhancer and / or additional excipients other than water.

[0456] In these embodiments, the weight ratio of polypeptide therapeutic agent to alginate oligomer in the solution or suspension is from about 1:0.5 to about 1:100, e.g., about 1:1, 1:2, 1:3, 1:4, 1:5, 1:6, 1:7, 1:8, 1:9, 1:10, 1:15, 1:20, 1:25, 1:30, 1:35, 1:40, 1:45, 1:50, 1:55, 1:60, 1:65, 1:70, 1:75 , 1:80, 1:85, 1:90, or 1:95 to about 1:100, or from about 1:0.5 to about 1:1, 1:2, 1:3, 1:4, 1:5, 1:6, 1:7, 1:8, 1:9, 1:10, 1:15, 1:20, 1:25, 1:30, 1:35, 1:40, 1:45, 1:50, 1:55, 1:60, 1:65, 1:70, 1:75, 1:80, 1:85, 1:90, or 1:95. Any range with endpoints formed from any of these values ​​is expressly contemplated.

[0457] In these embodiments, the weight ratio of the gastrointestinal permeation enhancer to the total amount of polypeptide therapeutic agent and alginate oligomer in the solution or suspension is from about 1:0.5 to about 1:100, e.g., about 1:1, 1:2, 1:3, 1:4, 1:5, 1:6, 1:7, 1:8, 1:9, 1:10, 1:15, 1:20, 1:25, 1:30, 1:35, 1:40, 1:45, 1:50, 1:55, 1:60, 1:65, 1:70, 1:80, 1:90, 1:100, 1:110, 1:120, 1:130, 1:140, 1:150, 1:160, 1:170, 1:180, 1:190, 1:200, 1:210, 1:220, 1:230, 1:240, 1:250, 1:260, 1:270, 1:280, 1:290, 1:300, 1:310, 1:320, 1:330, 1:340, 1:350, 1:360, 1:370, 1:380, 1:390, 1:410, 1:420, 1:430, 1:440, 1:450, 1:510, 1:520, 1:530, 1:540, 1:550, 1:560, 1:570, 1:580, 1:590, 1:610 1:70, 1:75, 1:80, 1:85, 1:90, or 1:95 to about 1:100, or about 1:0.5 to about 1:1, 1:2, 1:3, 1:4, 1:5, 1:6, 1:7, 1:8, 1:9, 1:10, 1:15, 1:20, 1:25, 1:30, 1:35, 1:40, 1:45, 1:50, 1:55, 1:60, 1:65, 1:70, 1:75, 1:80, 1:85, 1:90, or 1:95. Any range with endpoints formed from any of these values ​​is expressly contemplated.

[0458] A further example of a tablet for use in the present invention comprises a tablet comprising, in total, about 10% to about 50% weight / weight of an alginate oligomer and a polypeptide therapeutic agent, in total, about 50% to about 90% weight / weight of a gastrointestinal permeation enhancer; (i) Binders or Disintegrants (ii) lubricants, and / or (iii) fillers; and one or more excipients selected from the group consisting of: The tablet contains about 1% w / w or more polypeptide therapeutic agent, about 9% w / w or more alginic acid oligomer, and about 10% w / w or more gastrointestinal permeation enhancer.

[0459] Such tablets may be designed to dissolve essentially immediately in the gastric environment.

[0460] In certain embodiments, the tablet contains about 1% to about 20% weight / weight of the polypeptide therapeutic agent, e.g., about 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, or 19% to about 20% weight / weight, or about 1% to about 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, or 19% weight / weight. Any range, with endpoints formed from any of these values, is expressly contemplated.

[0461] In certain embodiments, the tablet contains about 9% to about 49% weight / weight alginate oligomer, for example, about 10, 15, 20, 25, 30, 35, 40, or 45% to about 49% weight / weight, or about 9% to about 10, 15, 20, 25, 30, 35, 40, or 45% weight / weight. Any range with endpoints formed from any of these values ​​is expressly contemplated.

[0462] In these embodiments, the weight ratio of polypeptide therapeutic agent to alginate oligomer in the tablet can be from about 1:0.25 to about 1:50, e.g., from about 1:0.5, 1:1, 1:2, 1:3, 1:4, 1:5, 1:6, 1:7, 1:8, 1:9, 1:10, 1:15, 1:20, 1:25, 1:30, 1:35, 1:40, or 1:45 to about 1:50, or from about 1:0.25 to about 1:0.5, 1:1, 1:2, 1:3, 1:4, 1:5, 1:6, 1:7, 1:8, 1:9, 1:10, 1:15, 1:20, 1:25, 1:30, 1:35, 1:40, 1:45, or 1:50. Any ranges with endpoints formed from any of these values ​​are expressly contemplated.

[0463] In certain embodiments, the tablet contains about 10% to about 70% weight / weight of the gastrointestinal permeation enhancer, e.g., about 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, or 65% to about 70% weight / weight, or about 10% to about 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, or 65% weight / weight. Any ranges with endpoints that may be formed from any of these values ​​are expressly contemplated.

[0464] In these embodiments, the weight ratio of gastrointestinal permeation enhancer to the combined amount of polypeptide therapeutic agent and alginate oligomer in the tablet can be from about 5:1 to about 1:9, e.g., from about 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, 1:6, 1:7, or 1:8 to about 1:9, or from 5:1 to about 4:1, 3:1, 2:1, 1:1, 1:2, 1:3, 1:4, 1:5, 1:6, 1:7, or about 1:8. Any ranges with endpoints formed from any of these values ​​are expressly contemplated.

[0465] In certain embodiments, the tablet contains about 10% to about 50% weight / weight filler, e.g., about 10, 15, 20, 25, 30, 35, 40, or 45% to about 50% weight / weight, or about 10% to about 15, 20, 25, 30, 35, 40, or 45% weight / weight. Any ranges with endpoints formed from any of these values ​​are expressly contemplated.

[0466] In certain embodiments, the tablet contains about 0.1% to about 20% weight / weight of lubricant, for example, about 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, or 19% to about 20% weight / weight, or about 0.1% to about 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, or 19% weight / weight. Any ranges with endpoints formed from any of these values ​​are expressly contemplated.

[0467] In certain embodiments, the tablet contains about 0.1% to about 20% weight / weight of binder or disintegrant, for example, about 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, or 19% to about 20% weight / weight, or about 0.1% to about 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, or 19% weight / weight. Any ranges with endpoints formed from any of these values ​​are expressly contemplated.

[0468] For the avoidance of doubt, in this aspect of the invention, the individual percentage amounts of each component sum to 100%. When the formulation is prepared, the components may hold solvent molecules, e.g., water molecules, in close association. For purposes of the present invention, residual solvent so associated is included proportionally in % wt / wt calculations and therefore does not affect the relative amounts of the components of the particles of the present invention.

[0469] "% wt / wt" (or "percent wt / wt") is a commonly used expression indicating the proportion of a solid composition made up of the compound in question. 1% wt / wt is equivalent to 1 gram of compound per 100 grams of solid composition, 2% wt / wt is equivalent to 2 grams of compound per 100 grams of solid composition, and so on. 1% wt / wt is also equivalent to 10 grams of compound per kilogram of solid composition.

[0470] "% wt / vol" (or "percent weight in volume") is a commonly used expression of the concentration of a solid solute in a liquid or semi-solid solution. 1% wt / vol is equal to 1 gram of solid per 100 ml of solvent, 2% wt / vol is equal to 2 g of solid per 100 ml of solvent, and so on. Thus, % wt / vol is expressed as g / 100 ml, grams per 100 milliliters, or g100ml -1 It is sometimes expressed as:

[0471] In these dosage form embodiments, the polypeptide therapeutic agent may be an incretin analog / mimetic, e.g., an agonist for the glucagon receptor, glucose-dependent insulinotropic polypeptide (GIP) receptor, and / or glucagon peptide-1 (GLP-1) receptor. In more preferred embodiments, the polypeptide is an agonist for the human forms of the above receptors.

[0472] Thus, in certain specific embodiments, the polypeptide therapeutic agent of the above-described formulations is dulaglutide (Trulicity), exenatide (Byetta), liraglutide (Victoza), semaglutide (Ozempic, Wegovy, and Ryvelsus), tirzepatide (Manjaro), albiglutide (Epelsan / Tanzeum), lixisenatide (Lyxumia / Adlyxin), polyethylene glycol loxenatide (Fulaimei), and LY3437943 (Retatortide), or structural analogs thereof.

[0473] The above-described formulations may be prepared by standard compounding techniques, so that one skilled in the art would be able to prepare the above-described formulations without undue burden. For example, the various ingredients may be dry-blended and pressed into tablet form or filled into capsules by standard means. Wet-blending of some or all of the ingredients may be performed prior to tableting or filling. In embodiments involving powders and / or pellets, various sizes may be used.

[0474] Powders / particles / pellets may be prepared by standard means such as milling, co-milling, spray drying, granulation (wet or dry), microdroplet gelation, microfluidic processing and the like. Powders and / or pellets may be administered neat or dissolved / suspended in a liquid, such as water or an aqueous solution (e.g., a solution of water and co-solvents and / or buffers as described above).

[0475] Formulations comprising layers and / or coatings may be prepared using conventional layering / coating techniques, such as spray coating or dip coating.

[0476] The invention will now be further described with reference to the following non-limiting examples. [Example]

[0477] Example 1 - Effect of gastrointestinal permeation enhancers on the diffusion of representative GLP-1 agonists in water

[0478] Diffusion of semaglutide, tirzepatide, and liraglutide in phosphate-buffered heavy water (DO) alone and in the presence of the gastrointestinal permeation enhancers SNAC or C10 1 H-NMR, 13 C was measured by HSQC and DOSY.

[0479] All samples were prepared in 10 mM phosphate buffer, pH 7.5, with 150 mM NaCl (PBS buffer) in 3 mm NMR tubes. Liraglutide, semaglutide, and tirzepatide (0.75–1.25 mM) were mixed with SNAC or C10. Measurements were performed at 25°C on a Bruker AVIIIHD 800 MHz system equipped with a 5 mm cryogenic CP-TCI. 1 H-NMR, 13 C HSQC and DOSY were recorded. Spectra were recorded using TopSpin 3.5 pL7 software (Bruker BioSpin) and processed and analyzed with TopSpin 4.0.7 software (Bruker BioSpin).

[0480] Semaglutide The DOSY and HQSC spectra of semaglutide and semaglutide with SNAC show that the addition of SNAC appears to slow the diffusion of semaglutide.

[0481] When SNAC was added 13 We observed changes in the chemical shifts of semaglutide in C-HSQC. Changes can also be seen in the 1D spectrum of semaglutide when increasing amounts of SNAC are added.

[0482] Tirzepatide The DOSY and HQSC spectra of tirzepatide and tirzepatide with SNAC and C10 show that the addition of SNAC increases the diffusion of tirzepatide, which increases further with the addition of C10.

[0483] When SNAC and C10 were added 13 Chemical shift changes of tirzepatide were observed in C-HSQC. Changes can also be seen in the 1D spectrum of tirzepatide when SNAC and C10 are added.

[0484] Liraglutide The DOSY and HQSC spectra of liraglutide and liraglutide with SNAC show that the diffusion of liraglutide is affected by SNAC, but to a lesser extent. The 1D spectrum of liraglutide also shows changes upon addition of C10.

[0485] Figure 1 shows a representative DOSY spectrum of tirzepatide (Tz) alone or in the presence of either SNAC or C10. The top three traces are, from top to bottom: tirzepatide light gray, tirzepatide + SNAC black, and tirzepatide + C10 dark gray.

[0486] Example 2 - Effect of gastrointestinal permeation enhancers and alginate oligomers on the transport of representative GLP-1 agonists through artificial mucus.

[0487] Time course of semaglutide diffusion through artificial mucus in a Transwell device in the presence of SNAC and / or Oligo G

[0488] Polycarbonate membrane Transwell plates (Costar) with 3 μm pore size and 96-well plates were used. Artificial mucus (Sigma) was 2.5% in Tris buffer, pH 7.2. 13 μL of mucus (approximately 900 μm thick) was added to the Transwell membrane. Semaglutide (SG) 200 μg / ml, Oligo G (OG-DP5-20, alginate oligomer with an average molecular weight of 3200 Da and 90-95% G residues) 6%, and SNAC 100 mM were added to the mucus layer. The samples were incubated at 37°C for 15, 30, 45, and 240 minutes before sampling from the acceptor chamber (bottom side). Samples were stored at -20°C until MS analysis. Each condition was performed in quadruplicate. Results are shown in Figure 2.

[0489] Diffusion of tirzepatide (TZ) through artificial mucus in a Transwell device after 60 min at 37°C, alone or in the presence of combinations of C10, SNAC and / or Oligo G (OG).

[0490] Polycarbonate Transwell plates (Costar) with 3 μm pore size and 24 wells. Artificial mucus (Sigma) was 2.5% in Tris buffer, pH 7.2. 30 μL of mucus (approximately 900 μm thick) was added to the Transwell membrane. Tirzepatide (TZ) 100 μg / ml, 6% oligo-G, and 100 mM each of SNAC and C10 were added to the mucus layer. The incubation was at 37°C for 60 minutes before sampling from the acceptor chamber (bottom side). Samples were stored at -20°C until MS analysis. Each condition was performed in quadruplicate. The results are shown in Figure 3.

[0491] Diffusion of liraglutide (LG) through artificial mucus in a Transwell device after 60 min at 37°C, alone or in the presence of a combination of SNAC and / or Oligo G (OG).

[0492] Polycarbonate Transwell plates (Costar) with 0.4 μm pore size and 24 wells. Artificial mucus (Sigma) was 2.5% in Tris buffer, pH 7.2. 10 μL of mucus (approximately 300 μm thick) was added on top of the Transwell membrane. Liraglutide (LG) 100 μg / ml, oligo-G (OG) 6%, and SNAC 100 mM were added to the mucus layer. The incubation was at 37°C for 60 minutes before sampling from the acceptor chamber (bottom side). Samples were stored at -20°C until ELISA analysis. Each condition was performed in quadruplicate. The results are shown in Figure 4.

[0493] Diffusion of semaglutide (SG) through artificial mucus in a Transwell device after 60 min at 37°C, alone or in the presence of SNAC and / or Oligo G (OG).

[0494] Polycarbonate Transwell plates (Costar) with 3 μm pore size and 14 wells. Artificial mucus (Sigma) was 2.5% in Tris buffer, pH 7.2. 10 μL of mucus (approximately 300 μm thick) was added on top of the Transwell membrane. Semaglutide (SG) 100 μg / ml, oligo G (OG) 6%, and SNAC 100 mM were added to the mucus layer. Incubation was at 37°C for 60 minutes before sampling from the acceptor chamber (bottom side). Samples were stored at -20°C until ELISA analysis. Each condition was run in quadruplicate. Results are shown in Figure 5.

[0495] Example 3 - Effect of gastrointestinal permeation enhancers on the systemic bioavailability of orally administered GLP-1 agonists and alginate oligomers

[0496] This study involved 50 Sprague-Dawley rats, 25 males and 25 females (2 spares), weighing approximately 300 g at the time of dosing. Animals were housed and maintained according to institutional procedures that conformed to animal welfare guidelines. Animals were fasted approximately 12 hours before dosing and until 2 hours after dosing.

[0497] Dose concentrations were prepared with a final gavage volume of 1.5 mL as defined in the table below. All doses were based on an assumed body weight of 300 grams for each animal. Formulations were prepared fresh on the day of dosing. Doses were administered directly into the stomach by gavage.

[0498] Blood samples (approximately 0.3 mL) were collected from the jugular vein into K2EDTA tubes at 0.25, 0.5, 0.75, 1, 1.5, 3, 6 and 8 hours after administration.

[0499] Samples were immediately inverted to ensure mixing with the anticoagulant and then placed on wet ice until processing to plasma. Plasma was generated by centrifugation (1500 × g, 10 min, +4°C) as soon as practical after blood collection, transferred to polypropylene Micronic tubes in a 96-well plate format, and stored in a freezer set to maintain temperatures below -65°C until analysis.

[0500] Samples were analyzed using appropriate LC-MS / MS and RGA2 methods for administered GLP-1 agonists, which are expected to detect intact and active GLP-1 agonists.

[0501] The results are shown in Figure 6 (semaglutide) and Figure 7 (tirzepatide). The AUC calculations are shown in the table below.

[0502] [Table 1]

[0503] [Table 2]

[0504] [Table 3]

[0505] [Table 4]

[0506] As can be seen, after oral administration of either the GLP-1 agonist semaglutide or tirzepatide with alginate oligomer and a gastrointestinal permeation enhancer, plasma concentrations of the GLP-1 agonist begin to rise almost immediately, reaching peak (Tz) or near-peak (80%, SG) levels essentially 15 minutes after administration. This indicates that uptake occurs primarily from the stomach, as the drug did not have time to pass through the stomach. Therefore, the increased bioavailability observed within 3 hours of administration can be attributed almost exclusively to enhanced uptake (absorption) from the stomach, since only a limited amount would have been able to reach the intestine over such a timescale. Such rapid uptake may provide an opportunity for improved oral dosing regimens.

[0507] The data also show a surprisingly increased systemic bioavailability (gastric absorption) for tirzepatide when administered orally with C10 (sodium caprate). The combination of oral semaglutide and SNAC had only a small effect on bioavailability at 3 and 8 hours, and what unnoticeable uptake did occur was slow. In contrast, tirzepatide and C10 showed rapid and significant absorption, although not as great as when these drugs were administered with alginate oligomers.

[0508] Data from bioavailability measurements in SD rats clearly show that alginate oligosaccharides can improve the bioavailability of orally administered polypeptides (represented by the GLP-1 agonists semaglutide and tirzepatide). This means that less polypeptide therapeutic agent can be administered to a subject to achieve the same pharmacological effect, which can help reduce manufacturing costs. The reduction in dose and subsequent reduction in formulation components can also reduce side effects caused by off-target exposure of formulation components, such as in the lower gastrointestinal tract.

[0509] Example 4 - Effect of alginate oligomers on the systemic bioavailability of orally administered semaglutide and SNAC

[0510] Introduction Patients with obesity and / or diabetes mellitus type II have been successfully treated with oral semaglutide together with sodium N-[8-(2-hydroxybenzoyl)amino]caprylate (also known as SNAC) when semaglutide is provided in tablet form. Such tablets are currently marketed by Novo Nordisk as Rybelsus. Buckley et al., 2018 ("Transcellular stomach absorption of a derivatized glucagon-like peptide-1 receptor agonist," Science Translational Medicine, Vol. 10, pp. 7047-7047) showed that systemic uptake occurs from the stomach and is a lengthy process that takes several hours (Figure 1B). Uptake appears to peak around 2 hours. This time course correlates with the dissolution rate of the tablet formulation used (Figure 1A). Absorption of semaglutide in gastric cell cultures also strongly depends on the concentration of SNAC present (Figure 2B), and the gradient of SNAC diffusing from the tablet was shown to be very steep. According to Buckley's data, suboptimal concentrations of SNAC, estimated to be present at 4 cm, were reached between 3 and 6 cm from the tablet (Figure 2C).

[0511] Therefore, the success of a patient's treatment with orally administered semaglutide is subject to complications from the presence or introduction of other gastric contents, such as food and beverages, and the patient's physical movements. These actions may dislodge the tablet from the stomach and / or interfere with the diffusion of tablet ingredients. Therefore, patients should not consume food within 30 minutes before or after administration of Ryvelsus tablets.

[0512] A means by which semaglutide could be absorbed from the stomach more quickly than Buckley tablets, for example in therapeutically effective amounts in the first 90 minutes after administration, would be advantageous. Experiments were carried out to determine whether administering alginate oligomer alongside semaglutide and SNAC could alter the absorption profile of semaglutide.

[0513] material and method The experiment was performed as described in Example 3. Dose concentrations were prepared as described below with a final gavage volume of 1.5 mL. All doses were based on an assumed weight of 300 grams for each animal. Formulations were prepared fresh on the day of dosing. Doses were administered directly into the stomach by gavage.

[0514] Semaglutide 6.67mg / kg High-dose Oligo-G 333 mg / kg Low-dose Oligo-G 100mg / kg Low dose SNAC 180mg / kg High dose SNAC 400mg / kg Oligo G (OG-DP5 to 20, alginate oligomer with an average molecular weight of 3200 Da and 90 to 95% G residues)

[0515] Samples were analysed using appropriate LC-MS / MS and RGA2 methods for semaglutide, which are expected to detect parent and active semaglutide.

[0516] Consideration The results are shown in Figures 8 to 10.

[0517] As can be seen, following oral administration of a liquid formulation of semaglutide and high levels of SNAC, consistent with the formulations of Ryvelsus tablets and Buckley tablets, semaglutide plasma concentrations remain unremarkable for up to 90 minutes. In contrast, the combination of Oligo G, semaglutide, and SNAC resulted in up to a 10-fold increase in semaglutide uptake (as measured by semaglutide plasma concentrations) during this time, with peak absorption occurring as early as 15 minutes. This suggests that orally administered solid formulations releasing semaglutide, Oligo G, and SNAC exhibit similar uptake profiles and may therefore deliver therapeutic levels of semaglutide to patients more quickly than Ryvelsus tablets. Such novel formulations would be less susceptible to food interference in a patient's stomach at the time of administration and to the consumption of food or beverages after administration.

[0518] Interestingly, uptake was enhanced during this period even when low levels of SNAC and Oligo G were used, suggesting that formulations containing reduced amounts of these active ingredients could be used clinically, thus conserving resources.

[0519] Example 5 - Effect of alginate oligomers on the systemic bioavailability of orally administered tirzepatide and C10

[0520] Introduction Tirzepatide has been successfully administered by subcutaneous injection to treat patients with obesity and / or type II diabetes mellitus. Oral administration has not been shown to be successful. While uptake of a similar therapeutic polypeptide, semaglutide, from the stomach following oral administration has been demonstrated using tablets that further contain the gastrointestinal permeation enhancer SNAC (marketed as Ryvelsus), successful treatment of patients with this orally administered tablet is susceptible to complications due to the presence or introduction of other stomach contents, such as food and beverages, and due to patient movement. These effects may dislodge the tablet from the stomach and / or interfere with the diffusion of tablet components. Therefore, patients should not consume food for 30 minutes before or after administration of Ryvelsus tablets.

[0521] If it can be shown that tirzepatide is absorbed from the stomach with the aid of a gastrointestinal permeation enhancer, a means by which therapeutically effective amounts of tirzepatide can be rapidly absorbed from the stomach, e.g., within the first 90 minutes after administration, would be advantageous. Experiments were conducted to determine whether administration of alginate oligomers together with tirzepatide and a prototypical gastrointestinal permeation enhancer, C10 (sodium caprate), would alter the absorption profile of tirzepatide.

[0522] material and method The experiment was performed as described in Example 3. Dose concentrations were prepared with a final gavage volume of 1.5 mL as defined below. All doses were based on an assumed weight of 300 grams per animal. Formulations were prepared fresh on the day of dosing. Doses were administered directly into the stomach by gastric gavage. Tirzepatide 7.65mg / kg Oligo G 333mg / kg C10 128.9 or 192 mg / kg Oligo G (OG-DP5 to 20, alginate oligomers with an average molecular weight of 3200 Da and 90-95% G residues)

[0523] Tirzepatide samples were analyzed using appropriate LC-MS / MS and RGA2 methods, which are expected to detect intact, active tirzepatide.

[0524] Consideration The results are shown in Figures 11 to 13.

[0525] As can be seen, after oral administration of a liquid formulation of tirzepatide with C10, tirzepatide concentrations remain unremarkable for up to 90 minutes. In contrast, combining oligo G with tirzepatide and C10 resulted in an 8-fold increase in tirzepatide uptake (measured by tirzepatide plasma concentrations) at specific time points during this period, and overall systemic bioavailability was almost 4-fold greater during this period, with uptake nearly peaking within 15 minutes and peaking by 30 minutes. This suggests that orally administered solid oral dosage forms that rapidly release tirzepatide, oligo G, and C10 will exhibit similar uptake profiles and thus be able to rapidly deliver therapeutic levels of tirzepatide to patients. Such novel formulations will be less susceptible to food disturbances in the patient's stomach at the time of administration and to the consumption of food or beverages after administration.

[0526] Example 6 - Effect of Alginate Oligomers on the Systemic Bioavailability of Orally Administered Retatortide (LY3437943) and C10

[0527] Sixteen male Sprague-Dawley rats weighing approximately 300 g at the time of dosing were used in this study. Animals were housed and maintained according to institutional protocols that conformed to animal welfare guidelines. Animals were fasted from 6 hours before dosing until 1.5 hours after dosing.

[0528] Dose concentrations were prepared with a final gavage volume of 0.75 mL as defined in the table below. All doses were based on an assumed body weight of 300 grams for each animal. Formulations were freshly prepared on the day of dosing. A single dose was administered directly to the stomach by gavage. Each formulation was administered to four rats.

[0529] [Table 5]

[0530] Blood samples (approximately 0.1 mL) were collected from the jugular vein into K2EDTA tubes 5, 10, 15, 30, 45, 60 and 90 minutes after administration.

[0531] The samples were immediately inverted to ensure mixing with the anticoagulant. The blood samples were then placed on ice after collection and centrifuged at approximately 3000xG for 5 minutes at 4°C. 20 μL of plasma was transferred to a separate low-binding protein tube, and the remaining plasma was transferred to a new tube. The tubes were frozen upright at approximately -20°C. The entire sample processing was completed within 30 minutes of sampling. The samples were then stored in a freezer set to maintain a temperature <-70°C until analysis.

[0532] Samples were analyzed using appropriate LC-MS / MS and RGA2 methods for retatortide, which are expected to detect intact and active retatortide.

[0533] The results are shown in Figures 14 and 15.

[0534] As can be seen, after oral administration of a liquid formulation of retatortide with C10, plasma concentrations of retatortide improved but remained unnoticeable for up to 90 minutes. In contrast, combining Oligo G with retatortide and C10 resulted in up to a five-fold increase in retatortide uptake (measured as retatortide plasma concentrations) at specific points during this period (specifically, 5 and 10 minutes). This suggests that orally administered, rapid-releasing solid oral dosage forms of retatortide, Oligo G, and C10 would exhibit similar uptake profiles and, therefore, be able to rapidly deliver therapeutic levels of retatortide to patients. Such novel formulations would be less susceptible to disturbance by food in a patient's stomach at the time of administration and to consumption of food or beverages after administration.

Claims

1. An alginate oligomer having 2 to 100 monomer residues for use in a method for increasing the systemic bioavailability of an orally, orally gavaged, nasogastric, or intragastrically administered polypeptide therapeutic agent, said method comprising administering said polypeptide therapeutic agent together with said alginate oligomer, and optionally a gastrointestinal permeation enhancer, to the stomach of a human or non-human animal as part of one or more dosage forms, said one or more dosage forms not bearing, in addition to said polypeptide therapeutic agent, said alginate oligomer, and, if used, said gastrointestinal permeation enhancer, a coating that provides substantial protection from the stomach environment.

2. 1. An alginate oligomer having 2 to 100 monomer residues for use in a method for the systemic treatment or prevention of a disease, condition, or complication thereof that is responsive to or prevented by a polypeptide therapeutic, said method comprising administering said polypeptide therapeutic, together with said alginate oligomer, and optionally a gastrointestinal permeation enhancer, to the stomach of a human or non-human subject having, suspected of having, or at risk of having said disease, condition, or complication thereof, as part of one or more dosage forms, said one or more dosage forms not bearing, in addition to said polypeptide therapeutic, said alginate oligomer, and, if used, said gastrointestinal permeation enhancer, a coating that provides substantial protection from the stomach environment.

3. 1. A dosage form adapted for delivery to the stomach of a human or non-human animal subject, said dosage form comprising: (i) a polypeptide therapeutic; and (ii) an alginic acid oligomer having 2 to 100 monomer residues; (iii) a gastrointestinal permeation enhancer; and wherein the dosage form does not have a coating that provides substantial protection from the gastric environment in addition to the polypeptide therapeutic agent, the alginate oligomer, and the gastrointestinal permeation enhancer.

4. 4. An alginate oligomer or dosage form for use according to any one of claims 1 to 3, wherein the alginate oligomer comprises: (i) 2 to 75, 2 to 50, 2 to 35, 2 to 30, 2 to 25, 2 to 22, 52 to 20, 2 to 18, 2 to 16, or 2 to 14; (i) 4 to 100, 4 to 75, 4 to 50, 4 to 35, 4 to 30, 4 to 25, 4 to 22, 4 to 20, 4 to 18, 4 to 16, or 4 to 14; (iii) 6 to 50, 6 to 35, 6 to 30, 6 to 25, 6 to 22, 6 to 20, 6 to 18, 6 to 16, or 6 to 14; (iv) 8 to 50, 8 to 35, 8 to 30, 8 to 25, 8 to 22, 8 to 20, 8 to 18, 8 to 16, or 8 to 14; or (v) a degree of polymerization (DP) or number average degree of polymerization (DP) of 10 to 50, 10 to 35, 10 to 30, 10 to 25, 10 to 22, 10 to 20, 10 to 18, or 10 to 14; ｎ ) an alginate oligomer having the following structure:

5. 5. The alginate oligomer or dosage form for use according to claim 1 , wherein the alginate oligomer is a dimer to 35-mer, a dimer to 30-mer, a trimer to 35-mer, a trimer to 28-mer, a 4-mer to 25-mer, a 5-mer to 20-mer, a 6-mer to 22-mer, an 8-mer to 20-mer, or a 10-mer to 15-mer.

6. 6. The alginate oligomer or dosage form for use according to claim 1 , wherein the alginate oligomer has at least 70%, at least 80%, at least 85%, at least 90% G, at least 95%, or 100% G residues.

7. 7. The alginate oligomer or dosage form for use according to claim 6, wherein at least 80% of the G residues are arranged in G-blocks.

8. 8. The alginate oligomer or dosage form for use according to claim 1 , wherein the alginate oligomer has at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, or 100% M residues.

9. 9. An alginate oligomer or dosage form for use according to claim 8, wherein at least 80% of the M residues are arranged in M ​​blocks.

10. 10. The alginate oligomer or dosage form for use according to any one of claims 1 to 9, wherein at least 70%, at least 80%, at least 85%, at least 90%, at least 95%, at least 99%, or 100% of the G and M residues in the alginate oligomer are arranged in MG blocks.

11. The alginate oligomer or dosage form for use according to any one of claims 1 to 10, wherein the polypeptide therapeutic agent is (i) to (iv) below or an analogue of (i) to (iv) below. (i) peptide hormones or growth factors (ii) cytokines (iii) enzyme or (iv) Blood factors

12. The polypeptide therapeutic agent may be selected from the group consisting of G-CSF, VEGF, EGF, HGF, PDGF, FGF, NGF, BDNF, neurotrophin-3, neurotrophin-4, TNF, IL-1, IL-6, IL-8, IL-10, IFN-γ, IFN-α, IFN-β, IFN-ε, IFN-κ and IFN-ω, Factor VII, Factor VIII and Factor IX, adrenocorticotropic hormone (ACTH), corticotropin-releasing factor, angiotensin, endothelin, calcitonin, insulin, glucagon, glucagon-like peptide-1, glucagon-like peptide-2, insulin-like growth factor, insulin-like growth factor-2, gastric inhibitory peptide, growth hormone-releasing factor, pituitary adenylate, and the like.

12. The alginate oligomer or dosage form for use according to claim 11, which is an acid cyclase-activating peptide, secretin, enterogastrin, somatostatin, somatotropin, somatomedin, parathyroid hormone, thrombopoietin, follicle-stimulating hormone, erythropoietin, gastrin, growth hormone, gonadotropin, hypothalamic releasing factor, prolactin, thyroid-stimulating hormone, endorphin, enkephalin, vasopressin, oxytocin, interferon, and analogs thereof, superoxide dismutase, asparaginase, arginase, arginine deaminase, adenosine deaminase, and ribonuclease, or an analog thereof.

13. 11. The alginate oligomer or formulation for use according to any one of claims 1 to 10, wherein the polypeptide therapeutic agent is or comprises an immunoglobulin amino acid sequence.

14. The polypeptide therapeutic agent is a therapeutic antibody, preferably alemtuzumab, bevacizumab, cetuximab, ofatumumab, panitumumab, rituximab, trastuzumab, ipilimumab, nivolumab, pembrolizumab, atezolizumab, avelumab, durvalumab, stimulimab, anifrolumab, bimekizumab, trolokinumab, evinacumab, aducanumab, ansuvimab, atortivimab, teprotumumab, eptinezumab, crizanlizumab, broecizumab, risankizumab , romosozumab, galcanezumab, erenumab, ibalizumab, emicizumab, benralizumab, ocrelizumab, sarilumab, dupilumab, bezlotoxumab, ixekizumab, alirocumab, vedolizumab, tocilizumab, canakinumab, infliximab, adalimumab, omalizumab, efalizumab, golimumab, ustekinumab, certolizumab pegol, ibritumomab, or tositumomab.

15. 11. The alginate oligomer or dosage form for use according to any one of claims 1 to 10, wherein the polypeptide therapeutic agent is a polypeptide antibiotic, preferably actinomycin, gramicidin, tylocidine, bleomycin, bacitracin, colistin and polymyxin B.

16. 11. The alginate oligomer or dosage form for use according to any one of claims 1 to 10, wherein the polypeptide therapeutic agent is an antimicrobial peptide or protein.

17. 11. The alginate oligomer or dosage form for use according to any one of claims 1 to 10, wherein the polypeptide therapeutic agent is an incretin peptide or an analogue / mimetic thereof, preferably glucagon-like protein 1, gastric inhibitory peptide (glucose-dependent insulinotropic polypeptide (GIP)), and / or exendin-4 peptide.

18. The polypeptide therapeutic agent includes dulaglutide, exenatide, liraglutide, semaglutide, tirzepatide, albiglutide, lixisenatide, polyethylene glycol loxenatide, retatortide (LY3437943), cotadutide, taspoglutide, langrenatide, veinaglutide, efpegrenatide, LY3502970, LY3537031, LY3493269, HM12525A / JNJ-64565111, MOD6030 / 1, SAR425899, MEDI0382, MK8521, ZP2929 / BI 456906, NN9709 / NNC0090-2746 / MAR709 / RG7697 / R06811135, SAR441255, C2816, ZP3022, NNC 9204-1177 (NN9277), LY3305677, JNJ-54728518, LY2944876 / TT-401, CPD86, SAR438335, ZP-I-98, ZP-DI-70, HM15211, NN9423 / MAR423, PB-719, and DD01.

19. The gastrointestinal permeation enhancer may be EDTA, citric acid, salicylic acid, oleic acid, lauric acid, acylcarnitine, acylcholine, and acylated amino acid, sodium glycocholate, sodium deoxycholate, sodium taurocholate, sodium dihydrofusidate, sodium glycodihydrofusidate, sodium glycolate, sodium lauryl sulfate, dioctyl sulfosuccinate sodium salt, sucrose fatty acid ester, lactose fatty acid ester, polysorbate, polyoxyethylene 11. The alginate oligomer or formulation for use according to any one of claims 1 to 10, wherein the alginate oligomer or formulation is selected from the group consisting of: cellulose acetate, cellulose acetate copolymer, cellulose acetate copolymer (cellulose acetate copolymer), ...

20. 1. A solid dosage form adapted for oral delivery to the stomach of a human or non-human animal subject, said solid dosage form comprising: (i) dulaglutide, exenatide, liraglutide, semaglutide, tirzepatide, albiglutide, lixisenatide, polyethylene glycol loxenatide, retatortide (LY3437943), cotadutide, taspoglutide, langrenatide, veinaglutide, efpegrenatide, LY3502970, LY3537031, LY3493269, HM12525A / JNJ-64565111, MOD6030 / 1, SAR425899, MEDI0382, MK8521, ZP2929 / BI 456906, NN9709 / NNC0090-2746 / MAR709 / RG7697 / R06811135, SAR441255, C2816, ZP3022, NNC 9204-1177 (NN9277), LY3305677, JNJ-54728518, LY2944876 / TT-401, CPD86, SAR438335, ZP-I-98, ZP-DI-70, HM15211, NN9423 / MAR423, PB-719, and DD01; (ii) an alginic acid oligomer having 2 to 100 monomer residues; wherein the solid dosage form does not have a coating that provides substantial protection from the gastric environment in addition to the polypeptide therapeutic agent and the alginate oligomer.

Images