Pharmaceutical compositions for peptide delivery

Abstract

To provide pharmaceutical compositions for delivery of peptide.SOLUTION: The present invention relates to a pharmaceutical composition comprising: a pharmaceutically effective amount of at least one peptide; and a pharmaceutically acceptable amount of a combination of: (a) at least one metal in form of any or a combination of a salt thereof and a complex thereof; and (b) at least one reducing agent. The at least one metal is selected from any or a combination of: vanadium, chromium and manganese. The combination of (a) at least one metal in form of any or a combination of a salt and a complex and (b) at least one reducing agent affords protection, at least in part, to the at least one peptide from proteolytic degradation upon ingestion thereof.SELECTED DRAWING: Figure 1

Description

Technical Field

[0001] The present disclosure generally relates to the field of pharmaceuticals. Specifically, the present invention includes a combination of a peptide, a metal salt / complex, and a reducing agent, and is at least partially a pharmaceutical composition that can protect the peptide from proteolytic degradation during ingestion.

Background Art

[0002] The background information in this specification includes information helpful for understanding the present invention. However, the background information in this specification does not assert that the present invention is well-known technology or is irrelevant to the protection scope of the present invention. Also, any materials or documents disclosed in the background information of this specification, whether explicit or implicit, do not represent that the content thereof indicates that the present invention is well-known technology.

[0003] Proteins and peptides have long been used in drugs such as therapeutic drugs and diagnostic drugs. Also, related technologies have continued to develop rapidly up to the present. However, since the development potential of proteins and peptides has not yet been fully understood, their application has remained limited to use by parenteral injection.

[0004] Oral administration is the best administration route that is simple and convenient. However, since peptides are decomposed after digestion in the digestive tract, it is difficult to be completely absorbed. Therefore, the low oral bioavailability of proteins and peptides is mainly due to enzymatic degradation in the digestive tract and low permeability of epithelial cells.

[0005] Regarding how to improve the problem of the low oral bioavailability of proteins and peptides, many methods have been proposed so far. For example, soybean trypsin inhibitor, aprotinin, Bowman-Birk type trypsin inhibitor, bacitracin, camostat mesilate, inhibitor amastatin (Renukuntla J et Examples include al., Int J Pharm. 2013, 447, 75-93; US Patent Application Publication US20070087957A1), in which various absorption enhancers and protease inhibitors are used. However, due to toxicity and numerous side effects, there are currently no commercially available additives that allow protease inhibitors to be used in polypeptide drugs for peptide delivery.

[0006] The following are some of the few specific examples of protease inhibitors that can be used for peptide delivery: (a) Soy (i.e., soy trypsin inhibitor): It is widely considered an allergen, and since 1980, the number of people suffering from soy allergies has gradually increased, leading to restrictions on its use (Moroz LA et al., N Engl J Med. 1980, 302, 1126~8; Foucard T et al., Allergy, 1999, 54, 261~5; Ramesh S, Clin Rev Allergy Immunol. 2008, 34, 217~30). Acute allergic reactions caused by soy include cough, sneezing, runny nose, hives, diarrhea, facial swelling, rapid breathing, tongue swelling, dysphagia, hypotension, excessive sweating, syncope, and anaphylactic shock, and can be fatal. (b) Bowman (i.e., Bowman-Birk type trypsin inhibitor): A soy derivative with high bioavailability when taken orally, and can have high bioavailability when taken orally without absorption enhancers. However, it has been reported that after taking Bowman orally, unnecessary systemic protease inhibitory effects (for example, inhibition of systemic serine proteases such as plasmin may increase the risk of thrombosis) may occur. Also, Bowman is It may also form antibodies against itself (Wan XS et al., Nutr Cancer, 2002, 43, 167~73). (c) Aprotinin: It is known to cause allergic reactions when used at a ratio of 1:200 for the first time (Mahdy AM et al., 2004, 93, 842~58), and aprotinin has been reported to be associated with an increased risk of acute renal failure, myocardial infarction, heart failure, stroke, and encephalopathy in patients with heart disease / surgery (Mangano DT et al., N Engl J Med, 2006, 354, 353~65).

[0007] Due to the potential health hazards associated with protease inhibitors, their use is generally avoided. In addition to the limitations mentioned above, protease inhibitors face challenges such as high manufacturing costs, heterogeneity, regulatory barriers, and difficulties in selective inhibition. Furthermore, the fact that effective activity is often only achieved at high doses makes widespread application difficult (Renukuntla J et al., Int J Pharm. 2013, 447). Other examples include bacitracin (antibiotic activity), camostat mesylate (effective in treating pancreatitis), or the inhibitor amastamin (antibacterial activity) (Renukuntla J et al., Int J Pharm. 2013, 447, 75-93; US-made). Other protease inhibitors, such as the published patent US20070087957A1, also have similar side effects to those described above.

[0008] European registered patent EP 3006045B1 is for trace elements (such as copper and zinc) and pharmaceutically acceptable substances. While possible reducing agent compositions are disclosed, these can be selectively supplemented with transmucosal absorption enhancers, and these compositions have been found to have beneficial and high bioavailability orally for a variety of peptide or protein-based drugs. However, because copper or zinc is related to mammalian metabolic pathways, the use of these compositions in long-term treatment may lead to negative interactions.

[0009] Therefore, in this field, there is a need for research and development of simple, safe, effective, and cost-effective pharmaceutical compositions that can deliver peptides while protecting them, at least partially, from proteolytic degradation during ingestion. This disclosure is in line with current and other needs and can reduce the technical shortcomings of conventional pharmaceutical compositions and their delivery. [Overview of the project] [Problems that the invention aims to solve]

[0010] This disclosure aims to provide a pharmaceutical composition that can overcome the drawbacks related to the compositions reported in the background art.

[0011] Another object of this disclosure is to provide a pharmaceutical composition that effectively delivers peptides.

[0012] Another object of this disclosure is to provide a pharmaceutical composition for delivering peptides orally.

[0013] Another object of this disclosure is to provide a pharmaceutical composition that protects ingested peptides from proteolytic degradation, at least in part.

[0014] Another object of this disclosure is to provide pharmaceutical compositions that increase the oral bioavailability of peptides.

[0015] Another object of this disclosure is to provide safe pharmaceutical compositions.

[0016] Another object of this disclosure is to provide a pharmaceutical composition that is economically efficient in manufacturing.

[0017] Another object of this disclosure is to provide pharmaceutical compositions that are easy to prepare.

[0018] Another object of this disclosure is to provide pharmaceutical compositions with a long shelf life.

[0019] This disclosure generally relates to the pharmaceutical field, and more specifically, to a pharmaceutical composition comprising a combination of a peptide, a metal salt / complex, and a reducing agent, which at least in part protects the peptide from proteolytic degradation upon ingestion.

[0020] One aspect of this disclosure is, At least one peptide in a pharmaceutically effective dose, and A pharmaceutically acceptable dose comprising (a) at least one metal in the form of a salt, a complex, or a combination thereof, and (b) at least one reducing agent, The combination of the at least one metal, which is selected from vanadium, chromium, and manganese or a combination thereof, and (a) the at least one metal in the form of a salt and / or a complex, or a combination thereof, and (b) the at least one reducing agent, provides a pharmaceutical composition that can at least partially protect the at least one peptide from proteolytic degradation upon ingestion.

[0021] In one embodiment, the at least one metal is vanadium, and the pharmaceutical composition contains either a vanadium salt or a vanadium complex, or a combination thereof, in an amount ranging from about 0.01 mg to about 15 mg per unit dose. In one embodiment, either the vanadium salt or the vanadium complex is independently selected from the group comprising vanadium(V) oxide, sodium vanadate, vanadium sulfate, vanadyl sulfate, vanadium biguanide, bis(maltolato)oxovanadium(IV), vanadium acetate, vanadyl picolinate, and vanadyl citrate. In one embodiment, the at least one metal is chromium, and the pharmaceutical composition contains either a chromium salt or a chromium complex, or a combination thereof, in an amount ranging from about 0.02 mg to about 0.5 mg per unit dose. In one embodiment, either the chromium salt or the chromium complex is independently selected from the group comprising chromium picolinate, chromium polynicotinate, chromium nicotinate, chromium chloride, and chromium acetate. In one embodiment, the at least one metal is manganese, and the pharmaceutical composition contains either a manganese salt or manganese complex, or a combination thereof, in an amount ranging from about 0.1 mg to about 10 mg per unit dose. In one embodiment, either the manganese salt or manganese complex is independently selected from the group including manganese gluconate, manganese sulfate, potassium permanganate, and manganese chloride.

[0022] In one embodiment, the at least one peptide has a molecular weight of 60 kDa or less. In one embodiment, the at least one peptide is insulin, insulin analog, insulin lispro, insulin peglispro, insulin aspart, insulin glulisine, insulin glargine, insulin detemir, protamine-containing intermediate-type (NPH) insulin, insulin degludec, B29K(N(ε)hexadecanedioyl-γ-L-Glu) A14E B25H desB30 human insulin, B29K(N(ε)octadecanedioyl-γ-L-Glu-OEG-OEG) desB30 human insulin, B29K(N(ε)octadecanedioyl-γ-L-Glu) A14E B25H desB30 human insulin, B29K(N(ε)eicosanedioyl-γ-L-Glu) A14E B25H desB30 human insulin, B29K(N(ε)octadecanedioyl-γ-L-Glu-OEG-OEG) A14E B25H desB30 human insulin, B29K(N(ε)eicosanedioyl-γ-L-Glu-OEG-OEG) A14E B25H desB3 0 Human insulin, B29K(N(ε)eicosanedioyl-γ-L-Glu-OEG-OEG) A14E B16H B25H desB30 Human insulin, B29K(N(ε)hexadecanedioyl-γ-L-Glu) A14E B16H B25H desB30 Human insulin, B29K(N(ε)eicosanedioyl-γ-L-Glu-OEG-OEG) A14E B16H B25H desB30 Human insulin, B29K(N(ε)octadecanedioyl) A14E B25H desB30 Human insulin, GLP-1, GLP-1 analog, acylated GLP-1 analog, diacylated GLP-1 analog, semaglutide, liraglutide, exenatide, lixisenatide,Selected from the group consisting of dual agonists of GLP-1 receptor and glucagon receptor, amylin, amylin analogs, pramlintide, somatostatin analogs, octreotide, lanreotide, pasireotide, goserelin, buserelin, leptin, leptin analogs, metreleptin, peptide YY, peptide YY analogs, glatiramer, leuprorelin, teriparatide, desmopressin, human growth hormone, human growth hormone analogs, glycopeptide antibiotics, glycosylated cyclic or polycyclic non-ribosomal peptide antibiotics, vancomycin, teicoplanin, telavancin, bleomycin, ramoplanin, decaplanin, bortezomib, cosyntropin, chorionic gonadotropin, menotropin, sermorelin, luteinizing hormone releasing hormone, somatropin, calcitonin, salmon calcitonin, pentagastrin, oxytocin, nesiritide, anakinra, enfuvirtide, pegvisomant, dornase alfa, repirin, anidulafungin, eptifibatide, interferon alfa con-1, interferon α-2a, interferon α-2b, interferon β-1a, interferon β-1b, interferon γ-1b, peginterferon α-2a, peginterferon α-2b, peginterferon β-1a, fibrinogen, vasopressin, aldesleukin, epoetin alfa, darbepoetin alfa, epoetin beta, epoetin delta, epoetin omega, epoetin zeta, filgrastim, interleukin-11, cyclosporine, glucagon, urokinase, viomycin, thyroid stimulating hormone releasing hormone, leucine enkephalin, methionine enkephalin, substance P, adrenocorticotropic hormone, parathyroid hormone, or pharmaceutically acceptable salts thereof.

[0023] In one embodiment, the at least one type of peptide and the at least one type of metal, which is in the form of a salt or a complex or a combination thereof, are present in a physically separated state in the pharmaceutical composition. In one embodiment, the at least one type of peptide and the at least one type of metal, which is in the form of a salt or a complex or a combination thereof, are present in separate compartments. In one embodiment, the pharmaceutical composition is in the form of a capsule-in-capsule dosage form or a tablet-in-capsule dosage form.

[0024] In one embodiment, the at least one reducing agent is selected from any one of ascorbic acid, reduced glutathione, cysteine, uric acid, reducing sugars, glyceraldehyde, α-tocopherol, vitamin A, α-lipoic acid, dihydro-α-lipoic acid, glucose, galactose, lactose, maltose, thiol-containing compounds, thiomers, and pharmaceutically acceptable salts thereof or a combination thereof. In one embodiment, the pharmaceutical composition contains the at least one reducing agent in an amount ranging from about 1 mg to about 1000 mg per unit dose.

[0025] In one embodiment, the pharmaceutical composition further contains at least one absorption enhancer, and the absorption enhancer is present in an amount ranging from about 10 mg to about 1000 mg per unit dose. In one embodiment, the pharmaceutical composition is prepared in the form of either an oral solid preparation or an oral liquid preparation, and when the pharmaceutical composition is prepared in the form of an oral liquid preparation, the pharmaceutical composition contains water in an amount less than about 5% (v / v).

[0026] Hereinafter, the content of the present disclosure will be described in detail based on the preferred embodiments and drawings of the present disclosure, and various members, technical features, aspects, and advantages of the present disclosure will be clearly disclosed. Members with the same reference numerals in the drawings are the same members.

Brief Description of the Drawings

[0027] [Figure 1]This graph, based on the embodiments of the specification, shows a comparison of insulin glargine (mU / L) concentrations and time from different formulations. [Modes for carrying out the invention]

[0028] Hereinafter, specific embodiments of this disclosure will be described in detail and clearly, along with the figures, in order to explain this disclosure in more detail. The embodiments are described in detail to clearly communicate this disclosure. However, the amount of detail provided is not intended to limit to anticipated variations of the embodiments. On the contrary, the intention is to encompass all variations, equivalents, and substitutes that fall within the spirit and scope of this disclosure as defined by the appended claims.

[0029] Each of the attached claims defines each of the inventions, and the infringing object is recognized as including equivalents of the various elements or limitations specified in the claims. Depending on the context of the following specification, the term “invention” as used herein may in some cases refer only to specific embodiments of this disclosure, and in other cases refer to one, but not all, of the subject matter described in the claims.

[0030] In this specification and in the claims, unless otherwise explicitly defined in the context, “a,” “an,” and “the” include the plural. Similarly, as used herein, unless otherwise explicitly defined in the context, the term “in” includes the meanings of “in” and “on.”

[0031] Unless otherwise stated or the context clearly contradicts the information herein, the methods described herein may be carried out in any appropriate sequence. The examples or illustrative terms, i.e., “for example,” in any or all of the embodiments provided herein are merely describing a good solution of the disclosure and do not limit the scope of protection of the disclosure. Nothing in this specification should be construed as making any element not described in the claims essential for the practice of the disclosure.

[0032] The various terms used herein are defined below. Unless otherwise defined below, terms used in the claims shall be interpreted in the broadest sense as they are reflected in the printed publications and issued patents at the time of filing by a person skilled in the art.

[0033] This disclosure generally relates to the pharmaceutical field, and more specifically, the present invention relates to a pharmaceutical composition comprising a combination of a peptide, a metal salt / complex, and a reducing agent, which at least partially protects the peptide from proteolytic degradation upon ingestion.

[0034] Serine proteases are universally present in eukaryotes and cleave peptide bonds, with serine, histidine, and aspartic acid being the three main catalytic residues. The serine proteases identified in this disclosure include trypsin, chymotrypsin, carboxypeptidase B, and aminopeptidase M. Serine proteases play a role in the body's physiological functions, particularly in digestion (protein degradation), i.e., the hydrolysis of peptide bonds and amino acids. This disclosure provides pharmaceutical compositions that, by comprising a combination of a peptide, a metal salt / complex, and a reducing agent, can at least partially protect peptides from protein degradation upon ingestion. The purpose is to do that.

[0035] Based on this, as one aspect of the present disclosure, a pharmaceutical composition is provided comprising a pharmaceutically effective dose of at least one peptide and a pharmaceutically acceptable dose of at least one metal in either the form of a salt or a complex, or a combination thereof, and (b) at least one reducing agent, wherein the at least one metal is selected from vanadium, chromium, and manganese, or a combination thereof, and the combination of (a) at least one metal in either the form of a salt or a complex, or a combination thereof, and (b) at least one reducing agent can at least partially protect the at least one peptide from proteolytic degradation upon ingestion.

[0036] In one embodiment, the at least one metal is vanadium, and the pharmaceutical composition contains either a vanadium salt or a vanadium complex, or a combination thereof, in an amount ranging from about 0.01 mg to about 15 mg per unit dose. In one embodiment, either the vanadium salt or the vanadium complex is independently selected from the group comprising vanadium(V) oxide, sodium vanadate, vanadium sulfate, vanadyl sulfate, vanadium biguanide, bis(maltolato)oxovanadium(IV), vanadium acetate, vanadyl picolinate, and vanadyl citrate. In one embodiment, the at least one metal is chromium, and the pharmaceutical composition contains either a chromium salt or a chromium complex, or a combination thereof, in an amount ranging from about 0.02 mg to about 0.5 mg per unit dose. In one embodiment, either the chromium salt or the chromium complex is independently selected from the group comprising chromium picolinate, chromium polynicotinate, chromium nicotinate, chromium chloride, and chromium acetate. In one embodiment, the at least one metal is manganese, and the pharmaceutical composition contains either a manganese salt or manganese complex, or a combination thereof, in an amount ranging from about 0.1 mg to about 10 mg per unit dose. In one embodiment, either the manganese salt or manganese complex is independently selected from the group including manganese gluconate, manganese sulfate, potassium permanganate, and manganese chloride.

[0037] In one embodiment, the at least one peptide has a molecular weight of 60 kDa or less. In one embodiment, the at least one peptide is insulin, insulin analog, insulin lispro, insulin peglispro, insulin aspart, insulin glulisine, insulin glargine, insulin detemir, protamine-containing intermediate-type (NPH) insulin, insulin degludec, B29K(N(ε)hexadecanedioyl-γ-L-Glu) A14E B25H desB30 human insulin, B29K(N(ε)octadecanedioyl-γ-L-Glu-OEG-OEG) desB30 human insulin, B29K(N(ε)octadecanedioyl-γ-L-Glu) A14E B25H desB30 human insulin, B29K(N(ε)eicosanedioyl-γ-L-Glu) A14E B25H desB30 human insulin, B29K(N(ε)octadecanedioyl-γ-L-Glu-OEG-OEG) A14E B25H desB30 human insulin, B29K (N(ε)eicosanedioyl-γ-L-Glu-OEG-OEG) A14E B25H desB30 human insulin, B29K (N(ε)eicosanedioyl-γ-L-Glu-OEG-OEG) A14E B16H B25H desB30 human insulin, B29K (N(ε)hexadecanedioyl-γ-L-Glu) A14E B16H B25H desB30 human insulin, B29K (N(ε)eicosanedioyl-γ-L-Glu-OEG-OEG) A14E B16H B25H desB30 human insulin, B29K (N(ε)octadecanedioyl) A14E B25H desB30 Human insulin, GLP-1, GLP-1 analogs, acylated GLP-1 analogs, diacylated GLP-1 analogs, semaglutide, liraglutide, exenatide, lixisenatide Dual agonists for GLP-1 receptors and glucagon receptors, amylin, amylin analogs, pramulintide, somatostatin analogs, octreotide, lanreotide, pasireotide, goserelin, buserelin, leptin, leptin analogs, metreleptin, peptide YY, peptide YY analogs, glatiramer, leuprorelin, teriparatide, desmopressin, human growth hormone, human growth hormone analogs, glycopeptides Antibiotics, glycosylated cyclic or polycyclic non-ribosomal peptide antibiotics, vancomycin, teicoplanin, teravancin, bleomycin, lamoplanin, decaplanin, bortezomib, cosyntropin, chorionic gonadotropin, menotropin, selmorelin, luteinizing hormone-releasing hormone, somatropin, calcitonin, salmon calcitonin, pentagastrin, oxytocin, nesiritide, anakinra, enfuvirtide, pegviso Selected from the group comprising mantle, dorunase alfa, repiridine, anidurafungin, eptifibatide, interferon alpha-1, interferon α-2a, interferon α-2b, interferon β-1a, interferon β-1b, interferon γ-1b, pegylated interferon α-2a, pegylated interferon α-2b, pegylated interferon β-1a, fibrinolysin, vasopressin, aldesleukin, epoetin alfa, darbepoetin alfa, epoetin beta, epoetin delta, epoetin omega, epoetin zeta, filgrastim, interleukin-11, cyclosporine, glucagon, urokinase, biomycin, thyroid-stimulating hormone-releasing hormone, leucine enkephalin, methionine enkephalin, substance P, adrenocorticotropic hormone, parathyroid hormone, or pharmaceutically acceptable salts thereof.

[0038] In one embodiment, the at least one peptide and the at least one metal, either in the form of a salt, a complex, or a combination thereof, are physically separated in the pharmaceutical composition. In one embodiment, the at least one peptide and the at least one metal, either in the form of a salt, a complex, or a combination thereof, are present in separate compartments. In one embodiment, the pharmaceutical composition exists in the form of a capsule-in-capsule or a tablet-in-capsule.

[0039] In one embodiment, the at least one reducing agent is selected from ascorbic acid, reduced glutathione, cysteine, uric acid, reducing sugars, glyceraldehyde, α-tocopherol, vitamin A, α-lipoic acid, dihydro-α-lipoic acid, glucose, galactose, lactose, maltose, thiol-containing compounds, thiomers, and pharmaceutically acceptable salts thereof, or a combination thereof. In one embodiment, the pharmaceutical composition contains at least one reducing agent in an amount ranging from about 1 mg to about 1000 mg per unit dose.

[0040] In one embodiment, the pharmaceutical composition further comprises at least one absorption enhancer, the absorption enhancer present in an amount ranging from about 10 mg to about 1000 mg per unit dose. In one embodiment, the pharmaceutical composition is prepared in the form of either an oral solid or an oral liquid, and if the pharmaceutical composition is prepared in the form of an oral liquid, the pharmaceutical composition contains water in an amount of less than about 5% (v / v).

[0041] In one embodiment, the peptide is any peptide or protein suitable for use as a therapeutic or diagnostic agent. In one embodiment, the peptide is a linear or cyclic peptide. In one embodiment, the peptide is a modified or derivatized peptide such as a PEGylated peptide, a fatty acid acylated peptide, or a di-fatty acid acylated peptide. The peptide may be histidine-free and / or cysteine-free. Typically, the peptide is preferably water-soluble, and particularly preferably has at least one serine protease cleavage site at a neutral pH (i.e., pH about 7), i.e., the peptide is serine protease-cleaved (especially trypsin, chymotrypsin, aminopeptides) It contains one or more amino acid residues that are suitable or easily cleaved by intestinal serine proteases such as -ase, carboxypeptidase, elastase and / or dipeptidylpeptidase 4.

[0042] In one embodiment, the peptide is insulin (preferably human insulin), but not limited to sustained-release basal insulin analogs, stabilized protease sustained-release basal insulin analogs, insulin lispro, insulin pegrispro, insulin derivatives such as A14E B25H B29K (N(eps) octadecanedi oil-gGlu-OEG-OEG), desB30 human insulin, insulin aspart, insulin glulisine, insulin glargine, insulin detemir, protamine-containing intermediate-release (NPH) insulin, insulin degludec, and insulin derivatives such as insulin analogs / derivatives disclosed in U.S. Patent Application Publication US20140056953A1, GLP-1, GLP-1 analogs (acylated GLP-1 analogs or diacylated GLP-1 analogs), semaglutide, liraglutide, exenatide, lixisenatide,Dual agonists for GLP-1 receptors and glucagon receptors, amylin, amylin analogs, pramulintide, somatostatin analogs (octreotide, lanreotide, or pasireotide), goserelin, buserelin, leptin, leptin analogs (metreleptin), peptide YY (PYY), PYY analogs, glatiramer (glatilamer acetate), leuprorelin, teriparatide, abaloparatide, tetracosactide, corticorelin, etelcalcetide, elcatonin, desmopressin, human growth hormone (hGH), human growth hormone analogs, glycopeptide antibiotics (vancomycin, teicoplanin, teravancin, bleomycin, lamopranin, decaplanin, etc., glycosylated cyclic (or polycyclic non-ribosomal peptide antibiotics), bortezomib, cosyntropin, chorionic gonadotropin, menotropin, selmorelin, luteinizing hormone-releasing hormone (LHRH, also called gonadotropin-releasing hormone), somatropin, calcitonin (salmon calcitonin), pentagastrinn, oxytocin, nesiritide, anakinra, enfuvirtide, pegvisomant, dorunase alfa, repiridine, anidurafungin, eptifibatide, interferon alfacon-1, interferon α-2a, interferon α-2b, interferon β-1a, interferon β-1b, interferon γ-1b, pegylated interferon α-2a interferon alfa-2a), pegylated interferon alfa-2b, pegylated interferon beta-1aThe insulin analogs, such as beta-1a), fibrinolysin, vasopressin, aldesleukin, epoetin alfa, darbepoetin alfa, epoetin beta, epoetin delta, epoetin omega, epoetin zeta, filgrastim, interleukin-11, cyclosporine, glucagon, urokinase, biomycin, thyroid-stimulating hormone-releasing hormone (TRH), leucine enkephalin, methionine enkephalin, substance P (CAS number 33507-63-0), adrenocorticotropic hormone (ACTH), parathyroid hormone (PTH), or any combination thereof of pharmaceutically acceptable salts. However, any other peptide molecules known or recognized by those skilled in the art may be used to serve the intended purposes without departing from the scope and spirit of the invention, as presented in this disclosure.

[0043] In one embodiment, the human target peptide is selected from either or a combination thereof of endogenous peptides such as insulin or glucagon. In preferred embodiments, a peptide-corresponding human isoform obtained by recombinant expression or chemical synthesis is used. However, any other human isoform peptide, known to or recognized by those skilled in the art, may be used to serve the intended purpose without departing from the scope and spirit of the invention, as presented in this disclosure.

[0044] In one embodiment, the peptide is an insulin analog. In one embodiment, the insulin analog is selected from insulin detemir, insulin glargine, insulin degludec, and other human, porcine, or fish-derived insulin analogs, or a combination thereof. However, any other insulin analogs / derivatives known or recognized by those skilled in the art may also be used to serve the intended purposes without departing from the scope and spirit of the invention, as presented herein.

[0045] In one embodiment, a mixture of two or more peptides may be used. In one embodiment, a mixture of human insulin and a GLP-1 agonist (e.g., liraglutide, semaglutide, exenatide, or lixisenatide) may be used. However, any mixture of two or more peptides (including the above-mentioned peptides) known or recognized by those skilled in the art may be used to achieve the intended purpose without departing from the scope and spirit of the invention, as presented in this disclosure.

[0046] In one embodiment, at least one peptide has a molecular weight of 60 kDa or less. In one embodiment, at least one peptide has a molecular weight of 40 kDa or less. In one embodiment, at least one peptide has a molecular weight of 30 kDa or less. In one embodiment, at least one peptide has a molecular weight of 20 kDa or less. In one embodiment, at least one peptide has a molecular weight of 10 kDa or less. In one embodiment, at least one peptide has a molecular weight of 300 kDa or more to 50 kDa or less. However, peptides having molecular weights within any range known or recognized to those skilled in the art can be used to achieve the intended purposes without departing from the scope and spirit of the invention, as presented in this disclosure.

[0047] In one embodiment, the molecular weight of at least one peptide can be measured by any method, as presented in this disclosure, to serve the intended purpose without departing from the scope and spirit of the invention, such as mass spectrometry (e.g., electrospray ionization mass spectrometry (ESI-MS) or matrix-assisted laser desorption / ionization mass spectrometry (MALDI-MA)), gel electrophoresis (e.g., sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE)), hydrodynamic methods (e.g., gel filtration chromatography or gradient sedimentation), or static light scattering (e.g., multi-angle light scattering detector (MALS)), or any other method known or recognized by those skilled in the art.

[0048] In one embodiment, at least one metal is vanadium, and the pharmaceutical composition comprises either a vanadium salt or a vanadium complex, or a combination thereof. In one embodiment, either a vanadium salt or a vanadium complex, or a combination thereof, is independently selected from the group comprising vanadium(IV), vanadium(V) such as vanadyl(V02) salt complexes, and vanadium as an anion in vanadate salts / complexes. In one embodiment, the vanadium salts and vanadium complexes are selected from any or a combination thereof, including vanadium(V) oxide, vanadium pentoxide, vanadium dioxide, sodium vanadate, vanadium sulfate, vanadyl sulfate, sodium metavanadate, vanadium tetrachloride, vanadium(V) oxychloride, vanadium oxytrichloride, vanadyl chloride, vanadium trichloride, ammonium vanadate, ammonium vanadium oxide, vanadium monosulfide, vanadium sulfide, vanadium(IV) chloride, vanadium biguanide, bis(maltolato)oxovanadium(IV) oxychloride, vanadium acetate, vanadyl picolinic acid, vanadyl citrate, etc. In one embodiment, the vanadium salts and vanadium complexes are vanadium(V). The use of vanadium(V) salts and complexes is due to their good water These are advantageous compared to vanadium(IV) salts and complexes due to their solubility and better oxidation state stability. In one embodiment, the vanadium salts and complexes are vanadium(IV) salts and / or complexes in which vanadium is part of the anion as vanadate or part of the cation as vanadyl. However, any vanadium salts and complexes or combinations thereof that are known or recognized by those skilled in the art can be used to serve the intended purposes without departing from the scope and spirit of the invention, as presented in this disclosure.

[0049] In one embodiment, the pharmaceutical composition contains either a vanadium salt or a vanadium complex, or a combination thereof, in an amount ranging from about 0.01 mg to about 15 mg per unit dose.

[0050] In one embodiment, the chromium salts and chromium complexes are preferably selected from chromium(III) salts and / or complexes. In one embodiment, any or a combination thereof of chromium salts and chromium complexes may be chromium picolinate, chromium chloride, chromium nicotinate, chromium polynicotinate, chromium acetate, trivalent chromium, high-chromium yeast, chromium pyridine-2-carboxylate, chromium tripicolinate, chromium 2-pyridinecarboxylate salt, tris(picolinate). Selected from sodium chromium, etc. In one embodiment, chromium salts and chromium complexes are It is more preferable to select from chromium picolinate, chromium polynicotinate, chromium nicotinate, chromium chloride, chromium acetate, etc. However, any chromium salts and complexes or combinations thereof that are known or recognized by those skilled in the art can be used to achieve the intended purposes of the invention as presented herein, without departing from the scope and spirit of the invention.

[0051] In one embodiment, the pharmaceutical composition contains either a salt of chromium or a complex of chromium, or a combination thereof, in an amount ranging from about 0.02 mg to about 0.5 mg per unit dose.

[0052] In one embodiment, either a manganese salt or manganese complex, or a combination thereof, is selected from manganese(II) salts and / or complexes, manganese(III) salts and / or complexes, or manganese salts and / or complexes as permanganates (V02). In one embodiment, either a manganese salt or manganese complex, or a combination thereof, is selected from manganese(II) sulfate (MnSO4), manganese(II) chloride (MnCl2), manganese(III) acetate, potassium permanganate, sodium permanganate, manganese gluconate, etc. In one embodiment, it is more preferable that any manganese salt or manganese complex is selected from manganese(III) salts and / or complexes. In one embodiment, the manganese(III) salt and / or complex is any or a combination thereof, such as manganese gluconate, manganese sulfate, manganese chloride, etc. However, any chromium salts and complexes or combinations thereof, as known or recognized by those skilled in the art, can be used to achieve the intended purposes of the present invention without departing from the scope and spirit of the invention, as presented herein.

[0053] In one embodiment, the pharmaceutical composition contains either a manganese salt or a manganese complex, or a combination thereof, in an amount ranging from about 0.01 mg to about 50 mg, preferably from about 0.1 mg to about 10 mg, per unit dose.

[0054] In one embodiment, any vanadium, chromium, manganese salts and complexes known or recognized by those skilled in the art can be used to serve the intended purpose of the invention without departing from the scope and spirit of the invention, as presented in this disclosure.

[0055] In one embodiment, vanadium salts and complexes enhance the oral bioavailability of peptides. Because the toxicity can be significantly increased, vanadium salts and complexes are preferred over chromium and manganese salts and complexes. In one embodiment, chromium salts and complexes are preferred over manganese salts and complexes. In one embodiment, the use of chromium salts and complexes is advantageous in that they have lower toxicity. In one embodiment, the use of manganese salts and complexes is superior to vanadium and chromium salts and complexes in that manganese salts and complexes are safe for humans even at high doses.

[0056] In one embodiment, the reducing agent is selected from any or a combination thereof from ascorbic acid (preferably ascorbates such as sodium ascorbate), reduced glutathione (GSH), cysteine, uric acid, reducing sugars (reducing monosaccharides such as glucose, glyceraldehyde, or galactose, or reducing disaccharides such as lactose or maltose), mannitol, α-tocopherol, vitamin A, α-lipoic acid, dihydro-α-lipoic acid (DHLA), thiol-containing compounds, thiomers (including thiomers disclosed in the literature Laffleur F et al., Future Med Chem, 2012, 4, 2205-16). In one embodiment, a mixture of two or more reducing agents may be used, with ascorbates and reduced glutathione being preferred. However, any reducing agent or combination thereof known or recognized by those skilled in the art can be used to achieve the intended purpose without departing from the scope and spirit of the invention, as presented in this disclosure.

[0057] In one embodiment, the pharmaceutical composition contains a reducing agent in an amount ranging from about 1.0 mg to about 1000 mg per unit dose, preferably from about 50 mg to about 500 mg.

[0058] In one embodiment, the pharmaceutical composition further comprises at least one absorption enhancer (or permeation enhancer). It should be noted that, as used interchangeably and synonymously in this disclosure, the terms “absorption enhancer” and “permeation enhancer” include the meanings of absorption enhancers and permeation enhancers known or recognized to those skilled in the art. In one embodiment, administration of at least one absorption enhancer or permeation enhancer improves or enhances the mucosal absorption rate of peptides in the gastrointestinal tract, particularly when the peptide size is large. In one embodiment, the at least one absorption enhancer or permeation enhancer is selected from either zwitterionic or nonionic absorption enhancers, or a combination thereof.In one embodiment, at least one accelerator is a C8-C20 alkanoylcarnitine (preferably lauroylcarnitine, myristoylcarnitine, or palmitoylcarnitine such as lauroylcarnitine chloride, myristoylcarnitine chloride, or palmitoylcarnitine chloride), salicylic acid (preferably a salicylate such as sodium salicylate), salicylic acid derivatives (e.g., 3-methoxysalicylic acid, 5-methoxysalicylic acid, or homovanylic acid), a C8-C20 alkanoic acid (preferably a C8-C20 alkanoate, more preferably a caprine, caprylate, myristicate, palmitate, or stearate such as sodium caprate, sodium caprylate, sodium myristate, sodium palmitate, or sodium stearate), citric acid (preferably a citrate such as sodium citrate), fat Acid-acylated amino acids (fatty acid-acylated amino acids disclosed in U.S. Patent Application Publication US20140056953A1, including sodium lauroyl alaninate, N-dodecanoyl-L-alanine, sodium lauroyl asparaginate, N-dodecanoyl-L-asparagine, lauroyl aspartic acid, N-dodecanoyl-L-aspartic acid, sodium lauroyl cysteinate, N-dodecanoyl-L-cysteine, sodium lauroyl Glutamic acid, N-dodecanoyl-L-glutamic acid, sodium lauroyl glutamate, N-dodecanoyl-L-glutamine, sodium lauroyl glycinate, N-dodecanoyl-L-glycine, sodium lauroyl histidinate, N-dodecanoyl-L-histidine, sodium lauroyl isoleucinate, N-dodecanoyl-L-isoleucine, sodium lauroyl leucinate, N-dodecanoyl-L-leucine. Sodium lauroyl methionine, N-dodecanoyl L-methionine, sodium lauroyl phenylalaninate, N-dodecanoyl-L-phenylalanine, sodium lauroyl proline, N-dodecanoyl L-proline, sodium lauroyl serine, N-dodecanoyl L-serine, sodium lauroyl threoninate, N-dodecanoyl-L-threonine, sodium lauroyl tryptophanate, N-dodecanoyl-L-tryptophan, sodium lauroyl tyrosinate, N-dodecanoyl-L-tyrosine, sodium Sodium lauroyl valinate, N-dodecanoyl-L-valine, sodium lauroyl sarcosinate, N-dodecanoyl-L-sarcosine, sodium capric alaninate, N-decanoyl-L-alanine, sodium capric asparaginate, N-decanoyl-L-asparagine, sodium capric aspartate, N-decanoyl-L-aspartic acid, sodium capric cysteinate, N-decanoyl-L-cysteine, sodium capric glutamate, N-decanoyl-L-glutamic acid, sodium capric glutamate Capricates, N-decanoyl-L-glutamine, sodium capric glycinate, N-decanoyl-L-glycine, sodium capric histidinate, N-decanoyl-L-histidine, sodium capric isoleucinate, N-decanoyl-L-isoleucine, sodium capric leucinate, N-decanoyl-L-leucine, sodium capric methioninate, N-decanoyl-L-methionine, sodium capric phenylalaninate, N-decanoyl-L-phenylalanine, sodium capric prolinate N-decanoyl-L-proline, sodium capric serinate, N-decanoyl-L-serine, sodium capric threoninate, N-decanoyl-L-threonine, sodium capric tryptophanate, N-decanoyl-L-tryptophan, sodium capric tyrosinate, N-decanoyl-L-tyrosine, sodium capric varianate, N-decanoyl-L-valine, sodium capric sarcosinate, N-decanoyl-L-sarcosine, sodium oleoyl sarcosinate, sodium N-decylleucine,Sodium stearoyl glutamate (Amisoft HS-11 P), sodium myristoyl glutamate (Amisoft MS-11), sodium lauroyl glutamate (Amisoft LS-11), sodium cocoyl glutamate (Amisoft CS-11), sodium cocoyl glycinate (Amilite GCS-11), sodium N-decylleucine, sodium cocoyl glycine, sodium cocoyl glutamate, sodium lauroyl alaninate, N-dodecanoyl L-alanine, sodium lauroyl asparaginate, 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-glutamic acid, sodium lauroyl glutamine, N-dodecanoyl L-glutamine, sodium lauroyl glycinate, N-dodecanoyl L-glycine, sodium lauroyl histidinate, N-dodecanoyl-L-histidine, sodium lauroyl isoleucinate, N-dodecanoyl L-isoleucine, sodium lauroyl leucinate, N-dodecanoyl L-leucine, sodium lauroyl methionine, N-dodecanoyl L-methionine, sodium lauroyl phenylalaninate, N-dodecanoyl-L-phenylalanine, sodium lauroyl prolineate, N-dodecanoyl-L-proline, sodium lauroyl serine, N-dodecanoyl-L-serine, sodium lauroyl threoninate, N-dodecanoyl-L-threonine, sodium lauroyl tryptophanate, N-dodecanoyl-L-tryptophan, sodium lauroyl tyrosinate, N-dodecanoyl -L-tyrosine, sodium lauroyl valinate, N-dodecanoyl L-valine, N-dodecanoyl-L-sarcosine, sodium capric alaninate, N-decanoyl-L-alanine, sodium capric asparaginate, N-decanoyl-L-asparagine, sodium capric aspartate, N-decanoyl-L-aspartic acid, sodium capric cysteinate, N-decanoyl-L-cysteine, sodium capric glutamate,N-decanoyl-L-glutamic acid, sodium, Sodium capric glutamine, N-decanoyl-L-glutamine, sodium capric glycinate, N-decanoyl-L-glycine, sodium capric histidinate, N-decanoyl-L-histidine, sodium capric isoleucinate, N-decanoyl-L-isoleucine, sodium capric leucinate, N-decanoyl-L-leucine, sodium capric methioninate, N-decanoyl-L-methionine, sodium capric phenylalaninate, N-decanoyl-L-phenylalanine, sodium capric proline, N-decanoyl-L-proline, sodium capric serine, N-decanoyl-L-serine, sodium capric threoninate, N-decanoyl-L-threonine, sodium pharmaceutically acceptable salts of the above compounds, such as sodium capric tryptophanate, N-decanoyl-L-tryptophan, sodium capric tyrosinate, N-decanoyl-L-tyrosine, sodium capric valinate, N-decanoyl-L-valine, sodium capric sarcosinate, sodium oleoyl sarcosinate, and C8-C20 alkanoyl sarcosinates (lauroyl sarcosinates such as sodium lauroyl sarcosinate), or compounds in which one of the 20 standard α-amino acids constituting proteins is acylated with a C8-C20 alkanoic acid, but not limited to these), alkylsaccharides (e.g., C8-C10 alkyl polysaccharides, specifically Multitrope TM1620-LQ-(MV), C1-C20 alkyl saccharides such as n-octyl-β-D-glucopyranoside or n-dodecyl-β-D-maltoside, cyclodextrin (α-cyclodextrin, β-cyclodextrin, γ-cyclodextrin, methyl-β-cyclodextrin, hydroxypropyl-β-cyclodextrin or sulfobutyl ether-β-cyclodextrin), sodium N-[8-(2-hydroxybenzoyl)amino]caprylate (SNAC), thiomer (including thiomer disclosed by Laffleur F et.al., Future Med Chem. 2012, 4, 2205-16), calcium chelating compounds (ethylenediaminetetraacetic acid (EDTA), glycol etherdiaminetetraacetic acid (EGTA), sodium citrate and polyacrylic acid), Kolliphor EL ( CAS no. 61791-12-6), chitosan, N,N,N-trimethylchitosan, benzalkonium chloride, bestatin, cetylpyridinium chloride, cetyltrimethylammonium bromide, C2-C20 alkanols (ethanol, decanol, lauryl alcohol, myristyl alcohol, or palmityl alcohol, etc.), C8-C20 alkenols (oleyl alcohol, etc.), C8-C20 alkenic acid (oleic acid, etc.), dextran sulfate, diethylene glycol monoethyl ether (transcutol), 1-dodecyl azacycloheptan-2-one (AzoneR), ethyl caprylate, glyceryl monolaurate, lysophosphatidyl Tidylcholine, menthol, C8-C20 alkylamines, C8-C20 alkenylamines (such as oleylamine), phosphatidylcholine, poloxamer, polyethylene glycol monolaurate, polyoxyethylene, polypropylene glycol monolaurate, polysorbate (polysorbate 80), deoxycholic acid (sodium deoxycholate), sodium glycocholate, sodium glycodeoxycholate, sodium lauryl sulfate (SDS), taurocholic acid (such as sodium taurocholate), taurodeoxycholic acid (taurodeoxycholic acid) The following are selected from any or a combination thereof: sodium phosphate, sucrose laurate, sulfoxides (C1-C10 alkyl-C1-C10 alkyl-sulfoxides such as decylmethyl sulfoxide or dimethyl sulfoxide), cyclopentadecalactone, 8-(N-2-hydroxy-5-chlorobenzoyl)-aminocaprylic acid (5-CNAC), dodecyl-2-N,N-dimethylaminopropionic acid (DDAIP), D-α-tocopheryl polyethylene glycol 1000 succinate (TPGS), and pharmaceutically acceptable salts of the above compounds. In one embodiment, a mixture of any two or more absorption enhancers, including the above absorption enhancers, may be used. However, any absorption enhancer or combination thereof known or recognized by those skilled in the art is not within the scope of the present invention as presented herein. It can be used to achieve its intended purpose without deviating from its intent.

[0059] In one embodiment, the pharmaceutical composition selectively contains an absorption enhancer or a permeation enhancer in an amount ranging from about 10 mg to about 1000 mg, preferably about 50 mg to about 500 mg, per unit dose.

[0060] In one embodiment, the composition of the pharmaceutical composition is configured such that when the pharmaceutical composition is added to 10 milliliters of a 5% hydrochloric acid solution, the acid is neutralized, resulting in a pH higher than approximately 6. In another embodiment, the composition of the pharmaceutical composition is configured such that when the pharmaceutical composition is added to 10 milliliters of an aqueous solution, it results in a pH in the range of 6 to 9.

[0061] In one embodiment, a pharmaceutical composition comprising at least one peptide having a molecular weight of 50 kDa or less in a pharmaceutically effective dose, at least one vanadium salt and / or a vanadium complex, or a composition thereof, at least one reducing agent, and optionally an absorption enhancer, can be administered orally and can protect at least one peptide from proteolytic degradation at least partially upon ingestion.

[0062] In one embodiment, a pharmaceutical composition comprising at least one peptide having a molecular weight of 50 kDa or less in a pharmaceutically effective dose, at least one chromium salt and / or chromium complex, at least one reducing agent, and optionally an absorption enhancer, can be administered orally and protect at least one peptide from proteolytic degradation at least partially upon ingestion.

[0063] In one embodiment, a pharmaceutical composition comprising at least one peptide having a molecular weight of 50 kDa or less in a pharmaceutically effective dose, at least one manganese salt and manganese complex, or a combination thereof, at least one reducing agent, and optionally an absorption enhancer, can be administered orally and can protect at least one peptide from proteolytic degradation at least partially upon ingestion.

[0064] In one embodiment, the pharmaceutical composition may further include any or a combination of one or more pharmaceutically acceptable excipients, such as carriers, diluents, fillers, disintegrants, lubricants, binders, colorants, pigments, stabilizers, preservatives, antioxidants, and / or solubility enhancers. In one embodiment, the pharmaceutical composition may further include one or more pharmaceutically acceptable additives, such as vitamin E, histidine, microcrystalline cellulose (MCC), mannitol, starch, sorbitol, and / or lactose. In one embodiment, the pharmaceutical composition may be modified by any technique known or recognized to those skilled in the art to achieve its intended purpose without departing from the scope and spirit of the invention, as presented in this disclosure.

[0065] In one embodiment, at least one solubility enhancer having a molecular weight range of about 200 to 5,000 Da is polyethylene glycol, ethylene glycol, propylene glycol, nonionic surfactant, tyroxapol, polysorbate 80, macrogol-15-hydroxystearate, phospholipid, lecithin, dimyristoylphosphatidylcholine, dipalmitoylphosphatidylcholine, distearoylphosphatidylcholine, cyclodextrin, α-cyclodextrin, β-cyclodextrin, γ-cyclodextrin, hydroxyethyl-β-cyclodextrin, hydroxypropyl-β-cyclodextrin, hydroxyethyl-γ-cyclodextrin, hydroxypropyl-γ-cyclodextrin, dihydroxypropyl-β-cyclodextrin, sulfobutyl ether-β-cyclodextrin, sulfobutyl ether-γ-cyclodextrin, glucosyl-α-cyclodextrin, glucosyl-β-cyclodextrin, diglucosyl The following are selected from or in combination thereof: 1-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, hydroxypropylcellulose, polyvinylpyrrolidone, vinyl acetate copolymer, vinylpyrrolidone, sodium lauryl sulfate, or sodium dioctyl sulfosuccinate. However, any dissolution accelerator or combination thereof known or recognized by those skilled in the art can be used to achieve the intended purpose without departing from the scope and spirit of the present invention, as presented in this disclosure.

[0066] In one embodiment, the pharmaceutical composition is preferably prepared for oral administration, more preferably for peroral administration. In one embodiment, at least one peptide, at least one metal in the form of a metal salt and / or a metal complex, or a combination thereof, at least one reducing agent, and optionally an absorption enhancer are administered orally.

[0067] In one embodiment, the dosage form of the oral pharmaceutical composition is selected from any or a combination thereof, including tablets (with or without coating), capsules (soft gelatin capsules, hard gelatin capsules, HPMC capsules, or HPMCP capsules), capsule-in-capsules, tablet-in-capsules, lozenges / troches, suppositories, solutions, emulsions, suspensions, syrups, elixirs, redissolvable powders and granules, dispersible powders and granules, medicinal gums, chewable tablets, effervescent tablets, multi-particle dosage forms, etc. However, any dosage form or combination thereof of a pharmaceutical composition known or recognized by those skilled in the art can be used to achieve the intended purpose without departing from the scope and spirit of the present invention, as presented in this disclosure.

[0068] In one embodiment, the tablets may contain, but are not limited to, microcrystalline cellulose, lactose, sodium citrate, calcium carbonate, calcium hydrogen phosphate, glycine, disintegrants (e.g., starch, preferably corn, potato, or tapioca starch), sodium starch glycolate, croscarmellose sodium, and specific complex silicates, as well as granulation accelerators (such as polyvinylpyrrolidone, hydroxypropyl methylcellulose (HPMC), hydroxypropyl cellulose (HPC), sucrose, gelatin, and acacia gum), and lubricants (such as magnesium stearate, stearic acid, glyceryl behenate, and talc powder). However, any excipient or combination thereof known or recognized by those skilled in the art may be used to achieve the intended purpose without departing from the scope and spirit of the invention, as presented in this disclosure.

[0069] In one embodiment, the capsule may contain, but is not limited to, any or a combination thereof of excipients such as lactose, starch, cellulose, or high molecular weight polyethylene glycol. However, any excipient or combination thereof known or recognized to those skilled in the art may be used to achieve the intended purpose without departing from the scope and spirit of the invention, as presented in this disclosure.

[0070] In one embodiment, the aqueous suspension and / or elixir is, but is not limited, any or a combination thereof, of excipients such as sweeteners or flavorings, colorants or dyes, emulsifiers and / or suspending agents, and diluents (such as water, ethanol, propylene glycol, and glycerin). Excipients may be included. However, any excipients or combinations thereof that are known or recognized by those skilled in the art can be used to achieve the intended purposes of the invention as presented herein, without departing from the scope and spirit of the invention.

[0071] In one embodiment, the pharmaceutical composition is prepared in either an oral solid dosage form or an oral liquid dosage form, and if the pharmaceutical composition is prepared in the form of an oral liquid dosage form, the pharmaceutical composition contains water in an amount of about less than 5% (v / v), preferably less than 3% (v / v), more preferably less than 1% (v / v), even more preferably less than 0.5% (v / v), even more preferably less than 0.1% (v / v), and even more preferably no water. In one embodiment, the oral liquid dosage form is particularly advantageous in that it can enhance storage stability. In one alternative embodiment, the oral liquid dosage form can be prepared quickly before administration and should not be stored for long periods.

[0072] The amounts of vanadium, chromium, and / or manganese used in the embodiments of this disclosure are far lower than the recommended daily intake of these trace elements and can therefore be considered safe. Furthermore, the combination of vanadium, chromium, and / or manganese with a reducing agent not only inhibits the action of serine proteases in the gastrointestinal tract but also does not produce systemic effects, resulting in a further improvement in safety compared to the protease inhibitors described in the background art. Moreover, compared to the protease inhibitors proposed for the oral delivery of peptide or protein drugs described in the background art, vanadium, chromium, and / or manganese, along with reducing agents such as ascorbic acid or reduced glutathione, can be provided at a considerably lower manufacturing cost.

[0073] Typically, a physician determines the optimal dosage for each individual patient. The specific dosage and frequency of administration of any given drug will vary for any given patient and depend on various factors, including the activity of the specific peptide or protein used, the metabolic stability and reaction time of the specific peptide or protein compound, as well as the patient's age, weight, health status, sex, diet, method and timing of administration, excretion rate, drug composition, severity in special circumstances, and the patient's treatment status. The precise dosage is ultimately at the discretion of the attending physician or veterinarian. Individuals or patients requiring treatment or prevention may include animals (non-human animals, etc.), vertebrates, mammals, rodents (guinea pigs, hamsters, rats, or mice, etc.), murids (mice, etc.), canids (dogs, etc.), felines (cats, etc.), pigs (swimming pigs, etc.), equids (horses, etc.), primates, monkeys (monkeys or apes, etc.), monkeys (gorillas, chimpanzees, orangutans, gibbons, etc.), or humans. The present invention may also consider treating animals that are economically or agriculturally important. Non-limiting examples of agriculturally important animals include sheep, cattle, and pigs, but cats and dogs may also be considered as economically important animals. The individual / patient is preferably a mammal, more preferably a human or non-human mammal (such as a guinea pig, hamster, rat, mouse, rabbit, dog, cat, horse, monkey, ape, marmoset, baboon, gorilla, chimpanzee, orangutan, gibbon, sheep, cattle, or pig), and even more preferably a human.

[0074] In one embodiment, at least one peptide, at least one metal in the form of a metal salt, a metal complex, or a combination thereof, at least one reducing agent, and optionally an absorption enhancer can be administered simultaneously, concomitantly, or sequentially. In one embodiment, sequential administration involves first administering at least one metal in the form of a metal salt, a metal complex, or a combination thereof, and at least one reducing agent, followed by at least one peptide and optionally an absorption enhancer (for example, at least about 5 minutes after the first administration, preferably about 5 minutes to about 3 hours after the first administration, and more preferably about 10 minutes to about 3 hours after the first administration). This is approximately 1 hour later, which is particularly advantageous when the peptide is insulin (human insulin). In one embodiment, at least one metal, either in the form of a metal salt or a complex or a combination thereof, at least one reducing agent, and optionally an absorption enhancer are administered, followed by the administration of the peptide (e.g., at least approximately 5 minutes after the first administration, preferably approximately 5 minutes to 3 hours after the first administration, and more preferably approximately 10 minutes to 1 hour after the first administration), which is also advantageous when the peptide is insulin (human insulin). In one embodiment, at least one metal is selected from vanadium, chromium, and manganese or a combination thereof.

[0075] In one embodiment, in simultaneous administration, at least one metal in the form of a metal salt and / or complex, or a combination thereof, and at least one reducing agent may be administered first, followed by at least one peptide and optionally an absorption enhancer, in the same pharmaceutical composition, two or more different / separated pharmaceutical compositions, or two or more different / separated compartments of the same pharmaceutical dosage form. In one embodiment, the at least one metal is vanadium, chromium, and manganese, or a combination thereof.

[0076] In one embodiment, at least one peptide and at least one metal, either in the form of a salt, a complex, or a combination thereof, are physically separated in the pharmaceutical composition.

[0077] In one embodiment, the pharmaceutical dosage form includes at least two separate, physically separated compartments (e.g., via a physical separation layer). In one embodiment, the pharmaceutical dosage form includes a physical separation layer between at least one peptide and at least one metal, either in the form of a salt, a complex, or a combination thereof. In one embodiment, in the pharmaceutical dosage form, at least one peptide is present only in the first compartment, and at least one metal, either in the form of a salt, a complex, or a combination thereof, is present only in the second compartment. In one embodiment, in the pharmaceutical dosage form, the reducing agent may be present in the first compartment, or in the second compartment, or in both the first and second compartments, or in the third compartment. In one embodiment, at least one metal is selected from vanadium, chromium, and manganese or a combination thereof.

[0078] In one embodiment, the pharmaceutical composition is in the form of a capsule-in-capsule or a tablet-in-capsule, and the pharmaceutical composition comprises: at least one peptide having a molecular weight of 50 kDa or less, present in a first compartment of the pharmaceutical dosage form; at least one metal, present in the form of a salt or complex of a metal, or a combination thereof, present in a second compartment of the dosage form; and a reducing agent present in the first and / or second compartments of the dosage form.

[0079] In one embodiment, the present invention provides a pharmaceutical dosage form (e.g., a multi-particle pharmaceutical dosage form) comprising: at least one peptide having a molecular weight of 60 kDa or less, present in a first compartment of the pharmaceutical dosage form; at least one reducing agent, present in a second compartment of the dosage form; and at least one metal, present in either the form of a metal salt, a complex, or a combination thereof, present in a third compartment of the dosage form. In one embodiment, the pharmaceutical dosage form is a capsule-in-capsule or tablet-in-capsule dosage form. In one embodiment, if the pharmaceutical dosage form is a capsule-in-capsule dosage form, the larger outer capsule (from which the contents are released first) contains at least one metal, present in either the form of a metal salt, a complex, or a combination thereof, and a reducing agent. The small inner capsule (from which the contents are released with a delay) contains a peptide. In one embodiment, at least one metal is selected from vanadium, chromium, and manganese, or a combination thereof. In one embodiment, the pharmaceutical dosage form is a modified-release dosage form (such as a dosage form with an enteric coating (e.g., capsule, multiparticle, or tablet)), a dosage form coated with Eudragit L30D55 ​​or Eudragit FS30D (e.g., capsule, multiparticle, or tablet), or an HPMCP capsule (commercially known as AR Cap). (R) One or a combination thereof, such as acid-resistant capsules (also known as) Selected from the combination. However, any other agent known or recognized by those skilled in the art deviates from the scope and spirit of the present invention as presented herein. [Examples]

[0080] Serine proteases: trypsin, chymotrypsin, carboxypeptidase B, and aminopeptidase M. Serine proteases are responsible for the proteolytic degradation of peptide bonds and amino acids. This experiment tested the oxidative inactivation of serine proteases under different combinations of metal ions and reducing agents.

[0081] Enzyme activity testing: Using an ultraviolet spectrometer at a specific wavelength, the enzymatic activity of different types of enzymes in specific substrates was measured. These test results were used as a negative control. The formula for calculating enzyme activity is as follows:

number

[0082] Enzyme inhibition test: The samples were incubated in the presence of inhibitors for each enzyme, and the results of this test were used as the positive control.

[0083] Incubation in the presence of metal ions and reducing agents: In a 96-well microtiter plate, combinations of enzymes, metal salts, and reducing agents were incubated with the enzymes for a set period of time to test the oxidative inactivation of the enzymes in the presence of the substrates. For enzyme inactivation in the presence of metal ions and reducing agents, the original enzyme activity, which was used as a negative control in the presence of the substrates, was compared with the positive control in the presence of the inhibitors.

[0084] pH measurement: A Hanna combination pH electrode was used to observe the change in pH value after incubation of the enzyme with different combinations of metal ions and reducing agents.

[0085] Zymography: Zymography was performed on the tested enzymes. Zymography is an electrophoretic technique that detects hydrolytic enzymes based on the enzyme's substrate repertoire. Specifically, when an acrylamide gel is prepared, the enzyme's substrate is incorporated into the gel of the layer being analyzed, allowing observation of the enzyme's digestion of the substrate.

[0086] Kit-based analysis: Enzyme inactivation was verified using a protease fluorescence assay kit and a trypsin activity assay kit.

[0087] material:

[0088] Table 1A below shows different metals and metal complexes (in some cases, a combination of one or more reducing agents). This is a test material for evaluating the inhibitory effect on serine protease activity (which can be achieved). [Table 1A] TIFF0007886581000003.tif154156

[0089] Table 1B below shows the inhibitory effects on serine proteases of different metal salt / metal complex and reducing agent combinations. [Table 1B]

[0090] Measurement of trypsin enzyme activity using Nα-benzoyl-L-arginine ethyl ester (BAEE)

[0091] A cold HCl solution of 200 units / mL trypsin and a 0.25 mM BAEE substrate solution were prepared and incubated. The above prepared solutions were inverted and stirred to form a reaction mixture. A blank solution (without enzyme) and a test solution (reaction mixture) were prepared. 253 Increase in absorbance The maximum linear velocity (ma) was measured and recorded (using a minimum of 4 data points per minute). Using ximum linear rate, the A of both the blank solution and the test solution 253 I got / minutes. The formula for calculating the 3ml trypsin test is as follows:

number

[0092] Measurement of trypsin inactivation in the presence of inhibitors: Trypsin 1 mM / L was incubated with specific (known) inhibitors (listed in Table 1A) and metal salt / reducing agent combinations (listed in Table 2 below). Inhibitor-induced inactivation was used as a positive control. [Table 2]

[0093] Determination of oxidative inactivation (activity and pH) of trypsin in the presence of metal ions and reducing agents.

[0094] Approximately 10 μl of trypsin was incubated with the respective concentrations of metal salts and reducing agents (listed in Table 2, serial numbers 1 to 31) in a buffer solution in a 96-well microtiter plate at 37°C for 5, 15, and 30 minutes (Table 3 below provides details on the use of specific combinations from those provided in Table 2, serial numbers 1 to 31). Then, 90 μl of substrate (listed in Table 1A) was added to each solution. Enzyme activity was measured spectrophotometrically using a microplate reader and compared to the original activity assay. pH was measured using a Hanna Combination pH electrode (reaction with the enzyme). The response was measured three times. Table 4 below shows the enzyme activity of trypsin after a specific period of time has elapsed following treatment with the combination of metal salt / complex and reducing agent being evaluated. [Table 3] [Table 4]

[0095] Tables 5A to 5C below show the pH values ​​of trypsin measured at different time intervals (i.e., 5 minutes, 15 minutes, and 30 minutes). [Table 5A] [Table 5B] [Table 5C]

[0096] Measurement of chymotrypsin enzyme activity using Ala-Ala-Phe-7-amide-4-methylcoumarin

[0097] Approximately 2 units of chymotrypsin were prepared in a cold HCl solution, along with a 1.18 mM substrate solution, a 2 M calcium chloride solution, and an 80 mM Tris HCl buffer. The substrate solution and calcium chloride solution were added to 3 mL of buffer solution, and the mixtures were inverted and stirred at approximately 25°C to prepare a blank reaction mixture and a test reaction mixture (Table 6). Furthermore, the HCl solution was added to the blank reaction mixture, and the enzyme solution was added to the test reaction mixture, and the mixtures were immediately inverted and stirred to prepare A 256 The increased absorbance was recorded for 3-5 minutes. At least four absorbances were recorded at intervals of more than 1 minute. The maximum linear rate of luminosity was used to determine the A of both the blank reaction mixture and the test reaction mixture. 256 I got / minutes. [Table 6]

number

[0098] Measurement of chymotrypsin inactivation in the presence of inhibitors: 1 mM / L chymotrypsin was incubated with specific known inhibitors (listed in Table 1 above) and combinations of metal salts and reducing agents (listed in Table 2 above). Inhibitor-inactivated solutions were used as positive controls.

[0099] Measurement of oxidative inactivation (activity and pH) of chymotrypsin in the presence of combinations of metal ions and reducing agents.

[0100] Approximately 10 μl of chymotrypsin was incubated with the respective concentrations of metal salt and reducing agent combinations (listed in Table 2, serial numbers 1 to 31) in buffer in a 96-well microtiter plate at 37°C for 5, 15, and 30 minutes (Table 3 below provides details on the use of specific combinations from those provided in Table 2, serial numbers 1 to 31). Then, 90 μl of substrate (listed in Table 1A) was added to each. Enzyme activity was measured spectrophotometrically using a microplate reader and compared to the original activity assay. pH was measured using a Hanna Combination pH electrode (the reaction with the enzyme was performed three times). Table 7 below shows the enzymatic activity of chymotrypsin after a specific period following treatment with the metal salt / complex and reducing agent combinations under evaluation. [Table 7]

[0101] Tables 8A to 8C below show the pH values ​​of chymotrypsin measured at different time intervals (i.e., 5 minutes, 15 minutes, and 30 minutes). [Table 8A] [Table 8B] [Table 8C]

[0102] Enzyme activity of carboxypeptidase B using hyprylarginine

[0103] A cold deionized aqueous solution of approximately 4 units of carboxypeptidase B and a 1 mg hyprylarginine solution in 25 nM Tris HCl buffer (pH 7.65) containing 100 nm of sodium chloride were prepared at approximately 25°C. According to Table 9, 3 mL of reaction mixtures (test and blank mixtures) were prepared at 25°C using the solutions prepared above. Calculation formula:

number

[0104] Measurement of carboxypeptidase B inactivation in the presence of inhibitors: Carboxypeptidase B (1 mM / L) was incubated with specific known inhibitors (listed in Table 1A above) and metal salt / reducing agent combinations (listed in Table 2 above). Inhibitor-inactivated samples were used as positive controls.

[0105] Measurement of oxidative inactivation (activity and pH) of carboxypeptidase B in the presence of metal ions and reducing agents.

[0106] For 200 μl of reaction mixture, add approximately 10 μl of carboxypeptidase to 96 wells. The metal salt and reducing agent combinations (listed in Table 2, serial numbers 1 to 31) were incubated in a buffer at 37°C for 5, 15, and 30 minutes at their respective concentrations (Table 3 below provides details on the use of specific combinations from those provided in Table 2, serial numbers 1 to 31), followed by the addition of 90 μl of substrate (listed in Table 1A) to each. Enzyme activity was measured spectrophotometrically using a microplate reader and compared to the original activity assay. pH was determined using the Hanna Combination pH scale. The enzyme activity was measured using an electrode (the reaction with the enzyme was performed three times). Table 10 below shows the enzyme activity of carboxypeptidase after a specific period following treatment with the metal salt / complex and reducing agent combinations under evaluation. [Table 10] TIFF0007886581000021.tif91154

[0107] Tables 11A to 11C below show the pH values ​​of carboxypeptidase measured at different time intervals (i.e., 5 minutes, 15 minutes, and 30 minutes). [Table 11A] [Table 11B] [Table 11C]

[0108] Enzyme activity of aminopeptidase M using L-leucine-P-nitroanilide

[0109] A 1 mM tricine solution (prepared in 100 mL of deionized water, Reagent A) and a 50 mM pure methanol solution of L-leucine-p-nitroanilide (Reagent B) were prepared. Approximately 10 mM L-leucine-p-nitroanilide solution (Reagent C) was prepared by adding 0.1 mL of Reagent B to 4.9 mL of Reagent A. A 200 mM tricine buffer solution (Reagent D) in deionized water and a 200 mM tricine buffer solution containing 0.05% (w / v) BSA, with a pH of 8.0 at 25°C (Reagent E) were prepared. 0.04 units / mL of aminopeptidase (enzyme solution, Reagent F) was prepared in Reagent E. Reagent C, (Leu-NA, 2. 0 ml of reagent (0 ml), reagent D (200 mM tricine buffer, 1.0 ml), and deionized water (7.0 ml) were pipetteed and mixed by swirling in the container to obtain the reaction cocktail (reagent G). Reagent G, reagent E, and reagent F were immediately inverted and stirred to prepare the test solution and blank solution as shown in Table 12, and after stirring and mixing, ΔA was set for about 5 minutes. 405 nm was recorded. And the maximum linear velocity (maximum A405 / min of the blank solution and test solution was obtained using a UM linear rate. Calculation formula:

number

[0110] Measurement of aminopeptidase inactivation in the presence of an inhibitor: 1 mM / L aminopeptidase Tidase was incubated with specific known inhibitors (listed in Table 1 above) and metal salt / reducing agent combinations (listed in Table 2 above). Inhibitor-inactivated tidases were used as positive controls.

[0111] Measurement of oxidative inactivation (activity and pH) of aminopeptidase in the presence of metal ions and reducing agents.

[0112] For 200 μl of reaction mixture, approximately 10 μl of aminopeptidase was incubated in a 96-well microtiter plate at 37°C with the respective concentrations of metal salt and reducing agent combinations (listed in Table 2, serial numbers 1 to 31) in buffer for 5, 15, and 30 minutes (Table 3 below provides details on the use of specific combinations from those provided in Table 2, serial numbers 1 to 31). Then, 90 μl of substrate (listed in Table 1A) was added to each. Enzyme activity was measured spectrophotometrically using a microplate reader and compared to the original activity assay. pH was measured using a Hanna Combination pH electrode (the reaction with the enzyme was performed three times). Table 13 below shows the enzyme activity of aminopeptidase after specific periods following treatment with the metal salt / complex and reducing agent combinations under evaluation. [Table 13] TIFF0007886581000028.tif90154

[0113] Tables 14A to 14C below show the pH values ​​of aminopeptidase measured at different time intervals (i.e., 5 minutes, 15 minutes, and 30 minutes). [Table 14A] [Table 14B] [Table 14C]

[0114] Based on the experiments conducted and the experiments described above, it could be concluded that the highest level of inhibition of enzyme activity (protein degradation) was observed by using the following combinations of reducing agents and metal salts: sodium ascorbate-vanadium oxide, benzohydroxamic acid-vanadium sulfate, mannitol-vanadium sulfate, uric acid-manganese gluconate, reduced glutathione-chromium chloride, and reduced glutathione-vanadium oxide.

[0115] Bioavailability test

[0116] To protect the peptide from protease degradation due to the acidic pH environment of the stomach, we prepared a capsule-in-capsule formulation in which the enteric-coated capsule protects the peptide from the gastric environment, the MIRA granules on the outside can inactivate proteolytic enzymes, and the permeabilis enhancer can promote / increase the absorption of the peptide into the epithelial membrane through intestinal permeability.

[0117] Preparation of capsule-in-capsule dosage form formulations

[0118] Preparation of granules containing reducing agents and metal salts

[0119] Tables 15A to 15F below show granular formulations (referred to here as MIRA granules) containing different reducing agents and metal salts, which were prepared to test their bioavailability. [Table 15A] [Table 15B] [Table 15C] [Table 15D] [Table 15E] [Table 15F]

[0120] Method for preparing MIRA granules

[0121] The following is the preparation process for MIRA granules.

[0122] A) Preparation of binder solution: A 0.3% w / v HPMC E-5 solution was prepared by dissolving 75.0 mg of HPMC E-5 in 25.0 mL of deionized (DI) water. B) Preparation of powder mixture: Each component was placed in a suitable container and thoroughly mixed using a plastic bag to ensure uniformity of the mixture. C) Preparation of granules: Granules were prepared by manually adding 2.0 mL of the binder solution dropwise (2 mL of binder was required to granulate 2.5 g m of the powder mixture). D) Drying of granules: The granules were placed in a hot air oven and dried at 40°C for 12 hours. E) Sieving of granules: The dried granules were sieved using a 40# mesh stainless steel wire mesh, and the sieved granules were collected in a suitable glass container and stored at room temperature. (Note that the temperature was 23°C and the humidity was 39% RH throughout the entire granulation process.)

[0123] Preparation of granules containing peptides

[0124] Tables 16A to 16I below show granular formulations (referred to here as PA granules) containing different peptides prepared for testing their bioavailability. [Table 16A] [Table 16B] [Table 16C] [Table 16D] [Table 16E] [Table 16F] [Table 16G] [Table 16H] [Table 16I]

[0125] Method for granulating peptide granules

[0126] A. Preparation of binder solution: A 0.3% w / v HPMC E-5 solution was prepared by dissolving 75.0 mg of HPMC E-5 in 25 mL of deionized (DI) water. B. Preparation of powder mixture: All components except the liquid excipient were accurately weighed and mixed for 5 minutes using a plastic bag. C. Addition of binder: The weighed liquid excipient and peptide were added to the 0.3% w / v HPMC E-5 binder solution. The resulting mixture was added dropwise little by little to perform wet granulation. D. Drying of granules: The granules prepared by granulation were placed on a silica gel bed in a vacuum desiccator and dried overnight. E. Sieving of granules: The dried granules were sieved using a 40# mesh stainless steel wire mesh, and the sieved granules were collected in a suitable glass container and stored at room temperature. (Throughout the entire granulation process, care was taken to maintain a temperature of 22°C and a humidity of 35% RH.)

[0127] Capsule filling

[0128] MIRA and peptide granules were manually filled into capsules using a weighing balance. Tables 17A and 17B below show the capsule sizes used for filling for the rat and canine studies, respectively. [Table 17A] [Table 17B]

[0129] Capsule packaging and storage

[0130] After packaging the capsules in a plastic bag, the packaged capsules were transferred to an HDPE (high-density polyethylene) container equipped with a silica bag for humidity control.

[0131] Preparation of placebo granules for batch testing

[0132] To visually understand the disintegration and release of granules from the capsules, placebo batches were prepared using sunset yellow and blue. Dried granules were filled into capsules of size 3, and disintegration time was measured using a guide disc on an electrolab. The results showed a disintegration time of 3 ± 1 minutes in water and phosphate buffer (pH 6.8) at 37 ± 0.2°C.

[0133] Dissolution test of placebo granules (capsule-in-capsule)

[0134] Outer capsule: Capsule size 0, yellow granules.

[0135] Inner capsule: Capsule size 4, blue granules.

[0136] Dissolution tests were performed at 37±0.5℃ in 900 mL of 0.1N HCl solution (for 2 hours) and in pH 6.8 phosphate buffer using a degradation test apparatus (Electrolab Mumbai).

[0137] The outer enteric-coated capsule remained perfectly intact in 0.1N HCl solution (i.e., stable in the gastric matrix), but visually observed disintegration began 3 minutes after being placed in pH 6.8 phosphate buffer.

[0138] The inner capsule began to disintegrate after 8 minutes and was completely dissolved within 13 minutes. Similarly, MIRA granules began to dissolve 3 minutes after exposure to alkaline phosphate buffer, and the peptide was completely dissolved within 13 minutes.

[0139] Capsule evaluation

[0140] Rat assays of insulin glargine, octreotide acetate, and teriparatide were performed. 50 mg / 30 mg granules (extracted from one capsule) were accurately weighed and dissolved in the mobile phase. The dispersion was sonicated for 5 minutes, filtered using a 0.22 μm syringe filter, and injected into HPLC. Calibration curves for all APIs were plotted using multiple dilutions with each mobile phase as recommended in USP2017. The assays were then performed. Next, the drug content was calculated using a calibration curve. [Table 18]

[0141] Stability testing of insulin glargine capsules [Table 19]

[0142] Capsule filling mode: manual filling; capsule size: 2; granule filling weight: same as for each formulation shown above.

[0143] Storage of capsules and granules: Temperature: 25±3℃. Humidity: 35±5%RH. Containers: HDPE 60 cc for capsules / clear glass vial with rubber stopper for granules.

[0144] Analysis results: From the formula (y = 30.977x - 292.26 obtained from HPLC data), it was found that the insulin glargine content in the formulations was 105.2% and 115.8% for batch I and batch II, respectively.

[0145] 75-day stability test: Both batches were stored at 25°C for 85 days and then analyzed again.

[0146] Analysis results: From the formula (y = 30.977x - 292.26 obtained from HPLC data), it was found that the insulin glargine content in the formulations was 97.25% and 91.63% for batch I and batch II, respectively, as can be seen in Table 20 below. [Table 20]

[0147] Teriparatide stability test [Table 21]

[0148] Capsule filling mode: manual filling; capsule size: 2; granule filling weight: same as for each formulation shown above.

[0149] Storage of capsules and granules: Temperature: 25±3℃. Humidity: 35±5%RH. Container: HDPE 60 cc for capsules / Clear glass vial with rubber stopper for granules.

[0150] Analysis results: From the formula (y = 18.924x - 22.539 obtained from HPLC data), it was found that the content of teriparatide acetate in the formulation was 98.02% and 106% for batch I and batch II, respectively.

[0151] 75-day stability test: Two batches were stored at 25°C for 75 days and then analyzed again.

[0152] From the formula (y = 18.924x - 22.539 obtained from HPLC data), the formulation The content of teriparatide acetate present in the material was found to be 92.88% for batch I and 89.76% for batch II, respectively. [Table 22]

[0153] Rat test measurement of leuprorelin acetate

[0154] Preparation of a leuprorelin acetate HPLC calibration curve: Approximately 112 mg and 104 mg of granules (equivalent to 100 μg) were accurately weighed and diluted using 1 mL of mobile phase; the theoretical concentration of leuprorelin acetate in this dispersion was found to be 100 μg / mL. The dispersion was vortexed for 2 minutes and then filtered using a 0.2 μm syringe. 20.0 μL of the filtrate was injected into the HPLC system to measure the peptide content. Analytical results: From the formula y = 42.67x - 76.447 obtained from HPLC for peptides, the % peptide content of leuprorelin acetate was (A) leuprorelin acetate + labrasol ALF granules (PA2): 97.45%, and (B) leuprorelin acetate + labrasol ALF + piperine granules (PA2 + 3): 105.11%.

[0155] Dissolution test of leuprorelin acetate capsules

[0156] Based on the analysis results, a dissolution test was performed on granular (PA2) formulations containing Labrasol ALF and leuprorelin acetate.

[0157] Setting 1: Use of dialysis membrane - Substrate: pH 6.80 phosphate buffer; Volume: 10 mL; Discharge volume: 400 μl; Stirring speed: 100 RPM; Packing granules: 140 mg (equivalent to one capsule); Dialysis membrane specifications: HIMEDIA LM395-30MT; Pore size: 25 nm; Average planar width: 29.31 mm; Average diameter: 17.5 mm. [Table 23]

[0158] Setting 2: Use of a rotating basket (USP TYPE 1) - Substrate: Phosphate buffer at pH 6.80; Volume: 25 mL; Discharge volume: 400 μl; Stirring speed: 100 RPM; Filling granules: 140 mg in gelatin capsules. [Table 24]

[0159] Setting 3: Use of a rotating basket (USP TYPE 1)

[0160] Substrate: pH 6.80 phosphate buffer; Volume: 30 mL; Discharge volume: 400 μl; Stirring speed: 500 RPM; Filling granules: 140 mg in gelatin capsules. [Table 25]

[0161] Setting 4: Use of a rotating basket (USP TYPE 1)

[0162] Substrate: Phosphate buffer at pH 6.80; Volume: 30 mL; Discharge volume: 400 μl; Stirring speed: 500 RPM; Filling granules: Hypromellose capsule (size 0) containing 140 mg. [Table 26]

[0163] Setting 5: Use of rotating basket (USP TYPE 1)

[0164] Substrate: pH 6.80 phosphate buffer; Volume: 30 mL; Discharge volume: 500 μl; Stirring speed: 100 RPM; Filling granules: 140 mg of granules were placed directly into the rotating basket without being prepared into capsules. [Table 27]

[0165] Setting 6: Use of a magnetic stirrer

[0166] Substrate: Phosphate buffer at pH 6.80; Volume: 30 mL; Discharge volume: 500 μl; Stirring speed: 200 RPM; Filling granules: 140 mg prepared in gelatin capsules (capsule size 2). [Table 28]

[0167] Setting 7: Use of a magnetic stirrer

[0168] Substrate: Phosphate buffer at pH 6.80; Volume: 30 mL; Discharge volume: 500 μl; Stirring speed: 200 RPM; Filling granules: 140 mg prepared in gelatin capsules (capsule size 2). [Table 29]

[0169] According to the data in Tables 24-29, it was noted that leuprorelin acetate did not elute from the granules and pass through the dialysis membrane (Setting 1). In Setting 2, a rotating basket (USP TYPE 1) was used, but the rotation of the basket did not generate enough rotational force to move / rotate the material in the substrate, causing the material to settle with the gelatin, ultimately affecting elution. In Setting 3, the RPM of the basket was increased (from 100 to 500), but even with increased RPM, the CDR% remained at 24.75%. In Setting 4, the RPM of the rotating basket was increased again, and hypromellose capsules were used instead of gelatin capsules, resulting in a CDR% of 29.97%; in Setting 5, the capsules were eliminated. The experiment was conducted using a rotating basket, and the CDR% was 90%, confirming that the material (gelatin / hypromellose) increased viscosity and delayed the elution of leuprorelin acetate from the granules. In settings 6 and 7, a magnetic stirrer was used to ensure good rotation of the substrate throughout the analysis, and the CDR% was 108.8% and 102.6%, respectively. All experiments were conducted at a setting of 37°C, but changes in temperature affect capsule disintegration. The disintegration time at 25°C was 12 minutes, and at 37°C it was less than 2 minutes.

[0170] Measurement of liraglutide sodium using HPLC

[0171] Test 1: Procedure: Approximately 10.0 mg each of granular PA2 (Table 16F) and granular PA3+4 (Table 16G) were diluted in 10.0 mL of HPLC diluent (DI water 10% ACN). The theoretical concentration of sodium liraglutide in this dispersion was found to be 74 μg / mL. The dispersion was sonicated for 5.0 minutes using an ultrasonic bath, filtered with a 0.2 μm syringe, and 20.0 μL of the filtrate was injected into the HPLC system to measure the peptide content. Analytical results: From the formula y = 79.283x - 571.72 obtained from HPLC for peptides, the content of the sodium liraglutide preparation with labrasol ALF (PA2) was 44.4%, and the content of the sodium liraglutide preparation with piperine and solutol HS15 (PA3+4) was 25.4%. (a) In a 2-minute vortex test, the analysis result for liraglutide sodium preparations containing piperine and Solutol HS15 was 67.39%; (b) In a 2-minute vortex test, the analysis result for liraglutide sodium granule preparations containing Labrasol ALF was 80.43%; (c) In a 5-minute vortex test, the analysis result for liraglutide sodium preparations containing piperine and Solutol HS15 was 95.66%; (d) In a 5-minute vortex test, the analysis result for liraglutide sodium granules containing Labrasol ALF was 96.99%.

[0172] Liraglutide dissolution test

[0173] Substrate: pH 6.80 phosphate buffer; Volume: 30 mL; Discharge volume: 500 μl; Stirring speed: 200 RPM; [Table 30] [Table 31]

[0174] Analysis results: The CDR% of liraglutide granules PA and PA3+4 were 112.85% and 93.64%, respectively. More than 50% of liraglutide sodium eluted within 5-7 minutes.

[0175] Quantitative analysis of glucose and insulin glargine content in STZ-induced diabetic rat plasma.

[0176] After administering an insulin glargine preparation to the midjejunum of STZ-induced diabetic rats, the glucose and insulin glargine content in their plasma was quantified.

[0177] Test formulation I: Insulin glargine (Lantus (R) )-Appearance: Pen-type refillable ink Injectable solution in a prefilled pen; concentration: 100 IU / ml; storage conditions: 2-8℃; dosage: 0.2 U / kg; route of administration: SC.

[0178] Test formulation II: Insulin glargine preparation - granule MIRA1 (reduced glutathione / chromium picolinate, Table 15A) + permeabilizer 1 (PA1, Table 1) (oral solution in TRIS buffer) - Dosage: 1.7U / animal; Route of administration: mid-jejunum.

[0179] Test formulation III: Insulin glargine preparations - PA2 (as shown in Table 16B) and MIRA2 (sodium ascorbate / vanadium oxide as shown in Table 15B) (oral solution in buffer) - Dosage: 1.7 U / animal; Route of administration: Mid-jejunum.

[0180] Test results: No rat deaths occurred with either subcutaneous injection or mid-jejunal administration. Clinical symptoms were normal. Regarding sampling timing, blood was collected during administration at 0, 20, 40, 60, 120, and 150 minutes post-administration. Approximately 100 μl of blood was collected from the posterior orbital sinus via puncture into a pre-filled Na-EDTA Eppendorf container. The blood was centrifuged at 5000 rpm for 5 minutes at 4°C to obtain plasma. Blood glucose levels were measured immediately after collection using a blood glucose meter. [Table 32] [Table 33]

[0181] ELISA test

[0182] Test material: Insulin glargine ELISA kit (Invitron Ltd, Cat. No. MBS495369).

[0183] Principle: This glargine ELISA is a two-site immunoassay that uses monoclonal antibodies immobilized in the wells of a microtiter plate and horseradish peroxidase (HRP)-labeled soluble antibodies for measurement. Plasma samples were incubated together in the wells of a microtiter plate, washed, and then HRP-conjugated antibody solution was added. Before measurement, unbound HRP-conjugated antibody was washed off and then a secondary incubation was performed. Enzyme substrates were added to each well of the microtiter plate, incubated for a period of time, and then further reagents were added to stop the reaction. The intensity of the color developed in each well was quantified using a microtiter plate reader set to record transmitted light at a wavelength of 450 nm (kit protocol: the protocol of catalog number MBS495369 recommended by the manufacturer was used).

[0184] Procedure: Before use, leave all kit contents and samples at room temperature. Fix the required number of coated strips to the plate holder. Store all strips not to be used immediately in a polyethylene sealed bag containing silica gel desiccant. Make sure that the remaining spaces on the plate holder are filled with uncoated strips to ensure uniform heat transfer during incubation. Pipette 100 μl of sample buffer into each well. Pipette 25 μl of standard or sample into each well. It is recommended to perform each test of the standard and sample in duplicate. Cover the plate holder and incubate at room temperature (18 - 22 °C) for 2 hours. Remove the plate holder cover and use an automatic plate holder washer to perform 3 washing cycles with cooled working strength washing buffer (300 μl per cycle). Pipette 100 μl of working strength conjugate antibody into each well. Cover with a plate sealer and incubate at 4 °C (2 - 8 °C) for 4 hours. Remove the plate sealer and use an automatic plate holder washer to perform 3 washing cycles with cooled working strength washing buffer. Next, add 100 μl of substrate solution to each well and incubate in the dark at room temperature (18 - 22 °C) for 15 minutes. Add 100 μl of stop solution to each well. Measure the light transmittance using a microtiter plate reader set at 450 nm and, if possible, using background subtraction measured at an OD of 620 / 650 nm. Figure 1 shows a graph depicting the concentration-time profile of insulin glargine (mU / L) from different formulations.

[0185] The test formulation insulin glargine - MIRA1 (reduced glutathione / picolinic acid chromium) + permeation enhancer 1 (PA1) showed a relative bioavailability of 9.25%, and the test formulations insulin glargine - PA2 and MIRA2 (sodium ascorbate / vanadium oxide)) showed a relative bioavailability of 28.86% .

[0186] Quantitative Analysis of the Content of Leuprorelin Acetate in Canine Plasma by ELISA Kit

[0187] Standard Test Preparation: Leuprorelin Acetate: LUPRODEX; Depot) - Concentration: Each vial contains 3.75 mg of leuprorelin acetate; Manufacturing Date: November 2017; Expiration Date: October 2020; Storage Environment: Store at room temperature (less than 25°C) without freezing; Number of Vials: 1 vial with diluent.

[0188] Test Preparation FB: MIRA5 (Table 15E) + PA2 (Table 16H) - Appearance: Hard gelatin capsule with white capsule cap and white capsule body; Concentration: Each capsule contains 300 mg and 1.25 mg of leuprorelin acetate; Manufacturing Date: April 21, 2018; Expiration Date: Not provided; Storage Environment: Store at room temperature (less than 25°C); Number of Capsules for Test Items: 70 capsules in 1 container.

[0189] Test Preparation H: MIRA2 (Table 15F) + PA2 + 3 (Table 16I) - Appearance: Hard gelatin capsule with white cap and white capsule body; Concentration: Each capsule contains 300 mg and 1.25 mg of leuprorelin acetate; Manufacturing Date: April 19, 2018; Expiration Date: None; Storage Environment: Store at room temperature (less than 25°C); Number of Capsules for Test Items: 70 capsules in 1 container.

Table 34

[0190] During the dosing period, dogs (Canidae; Canine species; Beagle) were fasted overnight (water was allowed) from 12 hours before dosing to 4 hours after dosing. 720 hours after dosing, the presence or absence of adverse events in dogs was observed. The body weight of each dog was measured and recorded before administration.

[0191] Sampling Time of Samples

[0192] 0 hours, 1 hour, 2 hours, 6 hours, 12 hours, 24 hours, 48 ​​hours, 72 hours, 96 hours At intervals of 120, 240, 312, 360, 480, and 720 hours (a total of 15 sampling time points), approximately 2 mL of blood samples were collected from the jugular vein of each dog that received subcutaneous and oral administration, using sample collection tubes coated and labeled with K2EDTA.

[0193] ELISA test conditions

[0194] Test material: Catalog number S-1174 (Des-Gly10, D-Leu6, Pro -NHEt9)-LHRH (leuprolide). Kit protocol: The protocol recommended by the manufacturer used (catalog number S1174).

[0195] Test results: The relative bioavailability of test formulation FB was 56.53%, while the relative bioavailability of test formulation H was 16%.

[0196] Determination of liraglutide content in canine plasma using an ELISA kit.

[0197] Test formulation I: Liraglutide - Appearance: Pre-filled injection solution in a pen-type syringe; Concentration: 6 mg / ml; Manufacturing date: February 2017; Expiration date: July 2019; Storage environment: 2-8°C; Dosage: 0.6 mg per dog; Route of administration: SC

[0198] Test formulation II (FA): MIRA5 (Table 15E) + PA2 (Table 16F) - Dosage: 12 mg (1 capsule) / dog; Route of administration: Oral.

[0199] Test formulation III(G): MIRA5 (Table 15E) + PA3 + 4 (Table 16G) - Dosage: 12 mg (1 capsule) / dog; Route of administration: Oral. [Table 35]

[0200] No dogs died after subcutaneous and oral administration of liraglutide. Clinical symptoms were normal. The body weight of each dog was measured and recorded before the study. During the treatment period, dogs (Canidae; breed; Beagle) were fasted at night from 12 hours before administration to 4 hours after administration (water was permitted). During the treatment period, blood samples were collected from dogs at 0, 20, 30, 60, 120, 180, 240, and 480 minutes after administration. 2 mL of blood sample was collected from the jugular vein into a sample collection tube coated and labeled with K2EDTA. Glucose was measured immediately after sampling using a blood glucose meter.

[0201] ELISA test conditions

[0202] Test material: ELISA kit (Krishgen BioSystems, Cat.No.KBI5020 Ver2.0).

[0203] Kit Protocol: The protocol recommended by the manufacturing company was used (Cat.No.KBI5020 Ver2.0). (1) Determine the wells for the diluted standards, blank control, and samples, and prepare 5 wells for the standards and 1 well for the blank control. Add 50 μL of the standards (for reading reagent preparation), blank control, and samples into the appropriate wells respectively. Immediately add 50 μL of liraglutide-biotin into each of the above wells. Gently shake the plate (using a plate shaker is recommended). Cover the plate with a plate cover. Incubate at 37 °C for 1 hour. Liraglutide-biotin may be turbid. After leaving it at room temperature, mix gently until the solution becomes homogeneous. (2) Aspirate the solution in the microplate, and then use a spray bottle, multi-channel pipette, manifold dispenser, and automatic washer to wash each well with 350 μL of 1× washing solution and let it stand for 1 - 2 minutes. Invert the plate onto absorbent paper to press and remove the remaining liquid. Repeat 3 times. After the last wash, remove all the remaining washing solution by aspiration or pouring, and invert the microplate onto absorbent paper. (3) Add 100 μL of streptavidin HRP sample into each well. Cover with a plate sealer and incubate at 37 °C for 30 minutes. (4) Repeat the aspiration / washing procedure 5 times in total as performed in step (2). (5) Add 90 μL of substrate solution into each well. Cover with a new plate sealer. Incubate at 37 °C for 10 - 20 minutes (not exceeding 30 minutes). Protect from light. The solution changes to blue by adding the substrate solution. (6) Add 50 μL of stop solution into each well. The solution changes to yellow by adding the stop solution. Tap the side of the plate to mix the mixture. If the color change of the solution is uneven, gently tap the microplate to ensure sufficient mixing of the solution. (7) Remove the water droplets or fingerprints on the bottom of the microplate and ensure that there are no bubbles on the liquid surface in the wells. Measure at 450 nm using a microplate reader.

[0204] The relative bioavailability of the test formulation FA (Mira5 and liraglutide + Labrasol) was found to be 3.82%, while the relative bioavailability of the test formulation G (Mira5 and liraglutide + Solutol HS15 + Piperine) was found to be 3.57%.

[0205] Quantitative analysis of octreotide content in rat plasma using ELISA

[0206] Test Formulation I: Octreotide - Appearance: Injectable solution; Concentration: 0.1 mg / ml; Storage conditions: 2-8°C; Dosage: 10 μg / kg; Route of administration: SC

[0207] Test formulation II: MIRA3 (Table 15C) + PA1 (Table 16C) administered to the distal small intestine (ileum) of anesthetized SD rats; Dosage: 144 μg / animal; Route of administration: Distal small intestine (ileum) injection. The rats were not fasted during the test. [Table 36]

[0208] No rat deaths occurred after subcutaneous and distal small intestine (ileum) administration of octreotide. Clinical symptoms were normal. During the administration period, blood samples were collected from dogs at 0, 7, 15, 30, 45, 60, and 90 minutes after administration. Blood samples were collected by puncturing the posterior orbital venous plexus, and approximately 100 μl of blood was collected in an Eppend container pre-filled with Na-EDTA. Blood samples were collected in a refractory tube. The blood samples were centrifuged at 4°C and 5000 rpm for 5 minutes to obtain plasma.

[0209] ELISA test

[0210] Test equipment: ELISA kit (Peninsula Laboratories International, Inc., Cat. No. S-1341.0001).

[0211] Kit Protocol: Use the protocol recommended by the manufacturer (Cat. No. S-1341.0001) - Add 25 μl of antiserum (in EIA buffer) to each well of the immunoplate. Add 25 μl of EIA buffer to the blank wells and incubate at room temperature for 1 hour. Add 50 μl each of the standard or sample (in diluent). Do not wash the plate before adding. Add 50 μl of diluent to the blank wells and incubate at room temperature for 2 hours. Sensitivity may be lower with short pre-incubation. Rehydrate Bt-tracer (in EIA buffer) and add 25 μl to each well and incubate overnight at 4°C. For better results, return to RT before operation. Wash the immunoplate 5 times with 300 μl / well of EIA buffer. Take sufficient care to avoid cross-contamination between wells during the first wash / dispense. In each washing cycle, quickly and gently move your wrist to empty the contents of the plate, then gently blot the top of the plate with a paper towel to dry. Dispense 300 μl of EIA buffer into each well, then gently shake the microplate for at least a few seconds. The washing procedure is essential. Add 100 μl of streptavidin HRP to each well. Tap or centrifuge the SAHRP tube to collect all the liquid components at the bottom, dilute the liquid inside to 1 / 200 (60 μl / 12 mL) in EIA buffer, and vortex. Add 100 μl to all wells, including the blank. Incubate at room temperature for 1 hour. Wash the immunoplate 5 times (see procedure (7)). Add 100 μl / well of TMB solution. Add to all wells, including the blank. Incubate at room temperature (usually 30-60 minutes). Read the blue color development at 650 nm and perform calculations using the data. Add 100 μl of 2N HCl to each well to stop the reaction. Read the absorbance at 450 nm within 10 minutes.

[0212] The bioavailability of the test formulation MIRA3 (uric acid: sodium vanadate) + PA1 was 0.41%.

[0213] Determination of teriparatide content in rat plasma by ELISA.

[0214] Test formulation I: Teriparatide; Appearance: Solution for injection; Concentration: 600 μg / 2.4 ml; Storage conditions: 2-8°C; Dosage: 10 μg / animal; Route of administration: SC

[0215] Test formulations II: MIRA4 (Table 15D) and PA1 (Table 16D) were administered to the distal small intestine (ileum) of anesthetized rats at a dose of 240 μg / animal; Dose: 240 μg / animal; Route of administration: Distal small intestine (ileum).

[0216] Test formulations III: MIRA1 (Table 15A) and PA3 (Table 16E) were administered to the distal small intestine (ileum) of anesthetized SD rats at a dose of 240 μg / animal; Dose: 240 μg / animal; Route of administration: Distal small intestine (ileum). [Table 37]

[0217] During the experiment, the animals were not fasted. No rat deaths occurred after administration of teriparatide subcutaneously and distally into the small intestine (ileum). Clinical symptoms were normal. During the administration period, blood samples were collected from dogs at 0, 7, 15, 30, 45, 60, and 90 minutes after administration. Blood samples were collected by puncturing the posterior orbital venous plexus, and approximately 100 μl of blood was collected in an Eppendorf container pre-filled with Na-EDTA. The blood samples were centrifuged at 5000 rpm for 5 minutes at 4°C to obtain plasma.

[0218] ELISA test

[0219] Test material: ELISA kit (Immutopics Cat. No. 60-3900)

[0220] Kit Protocol: Use the manufacturer's recommended protocol (Cat. No. 60-3900) - Place a sufficient number of streptavidin-coated strips into the holder and test the parathyroid hormone (PTH) standard, control, and unknown sample; pipette 150 μl of the standard, control, and sample into the designated or mapped wells, respectively. Freeze the standard and control samples immediately after use. Pipette 50 μl of working antibody diluent, consisting of 1 part HRP antibody and 1 part biotinylated antibody, into each well; cover with a plate sealer, cover with aluminum foil to protect from light; incubate the plate at room temperature for 3 hours with a horizontal rotor set to 180-200 rpm. Remove the aluminum foil and plate sealer and aspirate the contents of each well using an automated microplate washer. Wash each well 5 times, dispensing 350 μL of working wash diluent, and aspirate the contents of each well completely. A suitable aspiration device can be used; pipette 200 μl of ELISA HRP substrate into each well; cover the plate again with a plate sealer and aluminum foil. Incubate at room temperature for 3 hours with a horizontal rotor set to 180-200 rpm; remove the aluminum foil and plate sealer. Within 5 minutes, read the absorbance at 620 nm (see note) with a 0 pg / mL standard well as a blank using a microplate reader; immediately pipette 50 μl of ELISA stop solution into each well. Mix for 1 minute with a horizontal rotor; within 10 minutes, use a microplate reader to read the absorbance at 450 nm with 200 μl of substrate and 50 μl of stop solution as reagent blanks; if dual wavelength correction is possible, set the measurement wavelength to 450 nm and the reference wavelength to the absorbance used in step #9.

[0221] The relative bioavailability of the test formulations of MIRA4 (sodium ascorbate:manganese gluconate) and PA1 was 0.89%. The bioavailability of the test formulations of MIRA1 (reduced glutathione / chromium picolinate) and PA3 was also 0.89%.

[0222] While various embodiments of the present invention have been described above, other and further embodiments of the present invention can be made without departing from its basic scope. The scope of the present invention is determined by the claims. The present invention is not limited to the embodiments, modifications or examples described, and when combined with information and knowledge available to those skilled in the art, those skilled in the art may make the present invention. This also includes things that make it possible to manufacture and use them.

[0223] This disclosure provides a pharmaceutical composition that can overcome the drawbacks related to compositions reported in the background art.

[0224] This disclosure provides pharmaceutical compositions for effectively delivering peptides.

[0225] This disclosure provides pharmaceutical compositions for delivering peptides orally.

[0226] This disclosure provides a pharmaceutical composition that, when taken orally, protects peptides from proteolytic degradation, at least partially.

[0227] This disclosure provides pharmaceutical compositions that increase the bioavailability of peptides.

[0228] This disclosure provides a pharmaceutical composition that is safe.

[0229] This disclosure provides a pharmaceutical composition that is cost-effective, easy to prepare, and has a long shelf life.

Claims

1. At least one peptide in a pharmaceutically effective dose, and A pharmaceutically acceptable dose comprising (a) at least one metal in the form of a salt or complex or a combination thereof, and (b) at least one reducing agent, The at least one metal, in the form of either a salt or a complex or a combination thereof, is independently selected from the group consisting of manganese sulfate, manganese acetate, potassium permanganate, sodium permanganate, and manganese gluconate, and the combination of (a) at least one metal, in the form of either a salt or a complex or a combination thereof, and (b) at least one reducing agent can at least partially protect the at least one peptide from proteolytic degradation upon ingestion. A pharmaceutical composition that is a dosage form for oral pharmaceutical compositions.

2. The pharmaceutical composition according to claim 1, comprising either the salt and the complex or a combination thereof in an amount ranging from about 0.1 mg to about 10 mg per unit dose.

3. The pharmaceutical composition according to claim 2, wherein at least one metal, in the form of either the salt or complex or a combination thereof, is independently selected from the group consisting of manganese gluconate, manganese sulfate, and potassium permanganate.

4. The pharmaceutical composition according to claim 1, wherein the at least one peptide has a molecular weight of 60 kDa or less.

5. The aforementioned at least one peptide is insulin, insulin analog, insulin lispro, insulin pegrispro, insulin aspart, insulin glulisine, insulin glargine, insulin detemir, protamine-containing intermediate-type (NPH) insulin, insulin degludec, B29K (N(ε) hexadecanedioil-γ-L-Glu) A14E B25H desB30 human insulin, B29K (N(ε) octadecanedioil-γ-L-Glu-OEG-OEG) desB30 human insulin, B29K (N(ε) octadecanedioil-γ-L-Glu) A14E B25H desB30 human insulin, B29K (N(ε) eicosanedioil-γ-L-Glu) A14E B25H desB30 Human insulin, B29K (N(ε)octadecanedioyl-γ-L-Glu-OEG-OEG) A14E B25H desB30 Human insulin, B29K (N(ε)eicosanedioyl-γ-L-Glu-OEG-OEG) A14E B25H desB30 Human insulin, B29K (N(ε)eicosanedioyl-γ-L-Glu-OEG-OEG) A14E B16H B25H desB30 Human insulin, B29K (N(ε)hexadecanedioyl-γ-L-Glu) A14E B16H B25H desB30 Human insulin, B29K (N(ε)eicosanedioyl-γ-L-Glu-OEG-OEG) A14E B16H B25H desB30 Human insulin, B29K (N(ε)octadecanedioyl) A14E B25H desB30 Human insulin, GLP-1, GLP-1 analog, acylated GLP-1 analog, diacylated GLP-1 analog, semaglutide, liraglutide, exenatide, lixisenatide,Dual agonists for GLP-1 receptors and glucagon receptors, amylin, amylin analogs, pramulintide, somatostatin analogs, octreotide, lanreotide, pasireotide, goserelin, buserelin, leptin, leptin analogs, metreleptin, peptide YY, peptide YY analogs, glatiramer, leuprorelin, teriparatide, desmopressin, human growth hormone, human growth hormone analogs, glycopeptide antibiotics Antibiotics of the form of glycosylated, cyclic or polycyclic non-ribosomal peptides, vancomycin, teicoplanin, teravancin, bleomycin, lamoplanin, decaplanin, bortezomib, cosyntropin, chorionic gonadotropin, menotropin, selmorelin, luteinizing hormone-releasing hormone, somatropin, calcitonin, salmon calcitonin, pentagastrin, oxytocin, nesiritide, anakinra, enfuvirtide, pegvisomant, dorner The pharmaceutical composition according to claim 1, selected from the group consisting of zealpha, repiridine, anidurafungin, eptifibatide, interferon alphacon-1, interferon α-2a, interferon α-2b, interferon β-1a, interferon β-1b, interferon γ-1b, peginterferon α-2a, peginterferon α-2b, peginterferon β-1a, fibrinolysin, vasopressin, aldesleukin, epoetin alfa, darbepoetin alfa, epoetin beta, epoetin delta, epoetin omega, epoetin zeta, filgrastim, interleukin-11, cyclosporine, glucagon, urokinase, biomycin, thyroid-stimulating hormone-releasing hormone, leucine enkephalin, methionine enkephalin, substance P, adrenocorticotropic hormone, parathyroid hormone, or pharmaceutically acceptable salts thereof.

6. The pharmaceutical composition according to claim 1, wherein the at least one peptide and the at least one metal, which is either in the form of a salt or a complex or a combination thereof, are physically separated in the pharmaceutical composition.

7. The pharmaceutical composition according to claim 1, wherein the at least one peptide and the at least one metal, which is either in the form of a salt or a complex or a combination thereof, are present in the separated compartments.

8. The pharmaceutical composition according to claim 1, wherein the pharmaceutical composition exists in the form of a capsule-in-capsule or a tablet-in-capsule.

9. The pharmaceutical composition according to claim 1, wherein the at least one reducing agent is selected from any or a combination thereof of ascorbic acid, reduced glutathione, cysteine, uric acid, reducing sugars, glyceraldehyde, α-tocopherol, vitamin A, α-lipoic acid, dihydro-α-lipoic acid, glucose, galactose, lactose, maltose, thiol-containing compounds, thiomers, and pharmaceutically acceptable salts thereof.

10. The aforementioned pharmaceutical composition contains at least one reducing agent in an amount of approximately 1 mg to approximately 1000 mg per unit dose. The pharmaceutical composition according to claim 1, comprising an amount in the range of mg.

11. The pharmaceutical composition according to claim 1, further comprising at least one absorption enhancer, wherein the absorption enhancer is present in an amount ranging from about 10 mg to about 1000 mg per unit dose.

12. The pharmaceutical composition according to claim 1, wherein the pharmaceutical composition is prepared in the form of either an oral solid or an oral liquid, and if the pharmaceutical composition is prepared in the form of an oral liquid, the pharmaceutical composition contains water in an amount of less than 5% (v / v).

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