A composition for achieving oral absorption of a polypeptide

Abstract

The present application belongs to the field of biological medicine, and particularly relates to a polypeptide composition system for stable absorption in the gastrointestinal tract, and particularly discloses an oral polypeptide composition, which comprises a polypeptide molecule (A), a surfactant (B), and an amino acid single molecule or double molecule combination (C); the polypeptide drug for use as a medicine in the oral polypeptide composition has a molecular weight in the range of 0.1 kDa to 20 kDa. The action site of the composition system is the small intestine or the large intestine; the polypeptide composition of the present application has obvious improvement and increase in stability and absorbability in the gastrointestinal tract.

Description

TECHNICAL FIELD

[0001] The present application belongs to the field of biological medicine, and particularly relates to an oral polypeptide composition. BACKGROUND

[0002] Polypeptides are a class of compounds formed by the connection of multiple amino acids through peptide bonds, usually composed of 10-100 amino acid molecules, widely exist in biological bodies, and up to tens of thousands of polypeptides have been found in biological bodies so far. Polypeptides widely participate in and regulate the functional activities of systems, organs, tissues and cells in the body, and play an important role in life activities. Clinical treatment drugs of polypeptides show the advantages of high efficiency, selectivity and low toxicity, and have been widely used in many diseases including diabetes, cancer and metabolic diseases, and are more attractive than small molecule drugs (Fosgerau K, Hoffmann T Peptide therapeutics: current status and future directions. Drug Discov Today. 2015;20(1):122-128.). Compared with small molecule drugs, polypeptide drugs have the characteristics of high efficiency, low toxicity and strong specificity, and more than 80 polypeptide drugs have been approved for marketing by the US FDA. Polypeptides are macromolecules with good hydrophilicity, and have very poor permeability. In addition, the gastrointestinal tract is rich in proteases that degrade polypeptides, so most of the marketed polypeptide drugs are injections. Polypeptides are also easily degraded by peptidases in plasma, liver and kidney, have short half-lives, and have poor compliance for injection administration, which poses a challenge to the drug compliance of patients, especially for chronic patients who need long-term medication, which can cause long-term pain and injection aversion and other problems (Zizzari AT, et al. New perspectives in oral peptide delivery. Drug Discov Today. 2021;26(4):1097-1105.). In view of the above problems existing in the clinical use of polypeptide drugs, changing the administration mode and developing oral polypeptide drugs have become a promising solution. In addition, from the production point of view, compared with injection drugs, oral polypeptide drugs have greater production scale and lower production cost.

[0003] The gastrointestinal tract functions to digest carbohydrates, proteins and other nutrients into amino acids and monosaccharides, and to protect the body from pathogenic invasion. It is also the major obstacle for oral delivery of polypeptide drugs. The pH of gastric juice is 1-2 in the fasting state, and the strong acidic environment can induce polypeptide hydrolysis, deamination and oxidation, etc. to inactivate the polypeptide. In addition, the entire gastrointestinal tract is distributed with enzymes that can cleave polypeptides, including gastric protease secreted by gastric glands, pancreatic protease, chymotrypsin, carboxypeptidase and elastase secreted by pancreas, and aminopeptidase, endopeptidase and β-glutamine transpeptidase secreted by intestinal cells, etc. Guo et al. used bacitracin and leupeptin as protease inhibitors to improve the oral absorption of angiotensin 1 converting enzyme inhibitory peptide. The ideal polypeptide drug oral delivery system should be able to maintain the integrity of polypeptide molecules before reaching the absorption site, and release the drug at the target absorption site. The delivery system and the release site preferably have some interaction, and in addition, the system should be able to stay at the polypeptide release site to provide sufficient time and concentration gradient for the transmembrane transport of polypeptide drugs.

[0004] Based on the research progress of polypeptide drugs, there are currently two technical challenges for oral administration of polypeptide drugs. First, the stability of polypeptide drugs in the gastrointestinal tract. Due to the presence of proteases in the gastrointestinal tract, polypeptide drugs are usually quickly broken down into amino acids by proteases after oral administration, thereby losing drug activity (Wang J, et al. Toward oral delivery of biopharmaceuticals: an assessment of the gastrointestinal stability of 17 peptide drugs. Mol Pharm. 2015;12(3):966-973.). Proteases are ubiquitous in the gastrointestinal tract, and their species and concentration vary at different sites. The concentration and activity of proteases are higher near the front end, such as the stomach and duodenum, so polypeptide drugs are quickly broken down and inactivated after oral administration. In addition, the second challenge of oral administration of polypeptide drugs is the problem of gastrointestinal absorption efficiency. Generally speaking, drugs are absorbed from the gastrointestinal tract by penetrating into local venous vessels, completing the gastrointestinal absorption process, and entering the systemic circulation to exert efficacy. However, due to the structural characteristics of polypeptide drugs such as large molecular weight, it is difficult to penetrate through the mucus layer and epithelial cells of the gastrointestinal tract, so the gastrointestinal absorption rate of polypeptide drugs is very low (Verma S, Goand UK, Husain A, et al. Challenges of peptide and protein drug delivery by oral route: current strategies to improve the bioavailability. Drug Dev Res. 2021;82(7):927-944.).

[0005] Common polypeptide drug oral delivery technologies include:

[0006] Structural modification and cyclization of polypeptides: polypeptides are easily degraded by various enzymes in the body, but cyclization or covalent bonding of polypeptides can improve the enzyme stability of polypeptides, including stability in the whole body system, and prolong the half-life. Cyclosporine A (CsA) is a cyclic lipophilic 11 peptide, and its capsule (Sandimmune Ⓡ ) was approved for marketing by the US FDA in 1990, with an oral bioavailability of up to 25%-30%; the trans-isomer analog of CsA, voclosporin (Lupkynis Ⓡ) was approved for the treatment of lupus nephritis in January 2021, and its oral bioavailability is about 8% (HEO YA. Voclosporin: first approval [J]. Drugs, 2021, 81(5): 605-610.) Desmopressin acetate (Ddavp Ⓡ ) is a 9-peptide with certain hydrophilicity, which is a natural arginine vasopressin analogue obtained by deamination of the first amino acid and substitution of L-arginine with D-arginine at the 8th position. Although its oral bioavailability is only 0.08%-0.16%, it has low synthesis cost and high potency, so it can be taken orally (KOTTKE D, BURCKHAARDT BB, KNAAB TC, et al. Development and evaluation of a composite dosage form containing desmopressin acetate for buccal administration [J]. Int J Pharm X, 2021.3: 100082.). The polyethylene glycolated insulin analogue Tregopil insulin (IN-105) developed by the Indian Biocon company has completed phase III clinical trials, the growth hormone releasing peptide receptor agonist TZP-102 developed by the American Ocera company

[18] and SCY635 developed by the American Scynexis company for the treatment of hepatitis C infection have both completed phase II clinical trials. Although cyclization can improve stability, the large polar surface composed of a large number of amide bonds also limits the oral absorption of cyclic peptides. Nielsen et al. tested the physicochemical parameters and oral absorption of 125 cyclic peptides, most of which had poor bioavailability. How to improve the oral bioavailability of cyclic peptides needs further study.

[0007] Permeation enhancers (PEs) can transiently increase epithelial cell permeability and are one of the most widely used delivery strategies to improve oral absorption of polypeptides in clinic. Rybelsus® is the first oral glucagon-like peptide-1 (GLP-1) analogue to be marketed, which is achieved by the use of Eligen® technology developed by Emisphere Corporation, which uses a series of medium-chain fatty acid-based permeation enhancers, including N-2-hydroxybenzoyl-hexanoic acid sodium (SNAC), 8- [N-(2-hydroxy-5-chlorobenzoyl)amino] octanoic acid sodium (5-CNAC) and N-(4-salicyloyl chloride)-4-aminobutyrate (4-CNAB), etc. Among them, SNAC is the most widely used and is approved by the US FDA as a food additive. However, this technology chooses the stomach as the absorption site to solve the stability problem, and thus sacrifices the absorption efficiency of the drug. Buckley et al. determined that oral semaglutide is absorbed in the stomach by intragastric imaging, the effect of food on absorption, pylorus ligation and the distribution of semaglutide in the splenic and portal veins within 30 min after administration. SNAC (pKa1 = 4.5, pKa2 = 8.6) has strong buffering effect in the stomach, which can improve the enzyme stability of semaglutide by adjusting the pH, and also promote the penetration of semaglutide by promoting the depolymerization of semaglutide into monomers and interacting with lipid membranes. Since the stomach is not the main absorption organ compared to the intestine, and the effect of SNAC enhancer in the stomach is limited, the overall technology has the problem of low drug absorption rate. In the previous preclinical studies in beagle dogs and cynomolgus monkeys, the relative bioavailability of the oral formulation of this technology was in the range of 0.04-4.04%, and in subsequent human experimental studies, it only showed a bioavailability of 0.4-1%. Oral octreotide is the only oral somatostatin analogue for the treatment of acromegaly achieved by the TPE TM technology developed by Chiasma Corporation. TPE TMTechnology is to form a suspension by adding polypeptide, medium-chain fatty acid salt and polyvinylpyrrolidone (PVP) to hydrophobic medium, and deliver by enteric capsule. This technology has limited application range because it does not solve the technical challenge of stability, and only works for polypeptide drugs with very good stability, and is not a general delivery technology. In addition, because this technology only achieves the effect of transient penetration enhancement, the overall absorption rate is not high, and according to the results of human experiments, the absolute bioavailability is only 0.7% (Tuvia S, Atsmon J, Teichman SL, et al. Oral octreotide absorption in human subjects: comparable pharmacokinetics to parenteral octreotide and effective growth hormone suppression. J Clin Endocrinol Metab. 2012;97(7):2362-2369.). The sodium caprylate used in Mycapssa® can temporarily and reversibly open the paracellular tight junction, and the results of its phase III clinical study show good effectiveness and safety, with blood drug levels comparable to octreotide injection. Although the promotion of polypeptide oral absorption by PEs has been fully demonstrated, the spatiotemporal effect of polypeptides and PEs is often overlooked. After oral administration of semaglutide, high concentrations of semaglutide and SNAC can only be observed in the area directly below or on the surface of the tablet, and the absorption of semaglutide is also only in the local area where the tablet is located. The patent WO2013189988A1 published by Novo Nordisk shows that the formulation with faster SNAC release than semaglutide has higher bioavailability, which may be because SNAC dissolves first to form a buffer environment and enhance cell membrane permeability, reducing unnecessary loss of semaglutide. Therefore, the effect of gastrointestinal peristalsis on the co-delivery of polypeptides and PEs also affects the oral absorption of polypeptides. The movement of the empty stomach shows the characteristics of periodic and migratory complex movement, and the peristaltic ability of the duodenum and proximal jejunum is significantly higher than that of the ileum. Sladek et al. used insulin-related anionic polyelectrolyte nanoparticle complexes to encapsulate PEs and co-deliver them using enteric technology. In addition, for PEs that use the intercellular pathway, it may be necessary to appropriately increase the diffusion range of the formulation to open more tight junctions. In addition to the technologies that have been marketed, the GIPET® technology developed by Merrion uses sodium caprate as a PE, and in the phase II clinical study of the delivery of long-acting oral basal insulin "I338", it showed good blood glucose control ability and low risk of hypoglycemia.Oramed's POD technology. Oramed Pharma developed a Peptide Oral Delivery (POD) technology using EDTA as an absorption enhancer. The oral insulin developed by this technology failed in the clinical trial III. This technology selects the intestinal tract as the absorption site, overcomes the stability problem by adding protease inhibitor adjuvants (BBI or KTI), and overcomes the low absorption problem by using EDTA as an absorption enhancer. According to preclinical studies, this technology achieved a relative bioavailability of 5.41 ± 2.26% in beagle dog experiments (Li W-dw Y-Z, Zeng R, Greenberg-Shushlav Y, et al. Pharmacokinetic and pharmacodynamic profiles of orally, duodenally and subcutaneously delivered insulin in beagle canines. Paper presented at: American Diabetes Association (ADA); June 10-14, 2016; New Orleans, LA. 2016.). However, this technology has safety risks in the use ratio and long-term administration of chronic diseases due to the addition of protease inhibitors, which also partly explains the failure of the oral insulin developed by this technology in clinical trial III. Enteris Biopharma's Peptelligence technology. This technology selects the intestinal tract as the absorption site, and overcomes the stability problem by adding a large dose of organic acid (such as citric acid) to inhibit the protease activity in the local environment of the intestinal tract. In addition, this technology solves the absorption problem by adding a surfactant, L-lauroyl carnitine, to promote the intestinal permeation and absorption of polypeptides. The oral leuprolide developed by this technology showed an absolute bioavailability of 1.1 ± 0.18% in preclinical beagle dog studies, and has completed clinical trial II. However, the large amount of acid used by this technology to inhibit proteases in the intestinal tract (not less than 50 mg of dosage) has safety risks in long-term use. In addition, the overall absorption effect of this technology has not achieved high bioavailability.

[0008] (3) Nanotechnology: Nanoparticles (NPs) are solid particles with a diameter of 1-100 nm, which can not only protect polypeptides from being degraded by enzymes in the gastrointestinal tract, but also increase the transmembrane absorption of the intestinal epithelium, and can achieve targeted therapeutic effect through ligand modification. Oshadi Icp is a NPs delivery system developed by Oshadi Company for delivering insulin, proinsulin and C-peptide insulin composition, which uses pharmacologically inert hydrophobic silica nanoparticles as a carrier and can form a tight non-covalent bond with polypeptides and polysaccharides. The nanoparticles can protect polypeptides from the influence of gastrointestinal and brush border peptidases and promote the absorption of polypeptides. The published results of phase II clinical studies show that Oshadi Icp has good safety, tolerability and hypoglycemic effect in patients with type 1 diabetes. The hepatic directed vesicle insulin (HDV-I) developed by Diasome Company can directly deliver insulin to the liver after surface modification, with a particle size of <150 nm. After oral administration of HDV-I, it is taken up in the intestinal tract through the hepatic portal vein, which can achieve normal physiological response of insulin. The completed phase II clinical study results show that HDV-I has significant hypoglycemic effect in oral glucose tolerance test and diabetic meal, and phase III clinical study is in preparation. With the continuous progress of the physicochemical characterization technology and imaging technology of nanostructure, the biocompatibility, reproducibility and scalable production of NPs are still the difficulties of current research, and the core technology needs to be updated.

[0009] (4) Drug-in-Device: Drug-in-Device is a new direction for oral delivery of polypeptides, which can be applied to the delivery of a variety of different peptides, and has lower requirements for the size, stability and hydrophobicity of polypeptide molecules. Drug-in-Device includes microneedles, enteric patches, microcontainers, etc., among which microneedles are the most promising technology. There are no pain receptors in the intestines, so painless drug delivery can be achieved. Drug-in-Device can protect polypeptides from being degraded by enzymes in the gastrointestinal tract on the one hand, and achieve one-way co-delivery of polypeptides and PEs in time and space on the other hand. The self-orienting millimeter-scale applicator (SOMA) developed by MIT and Novo Nordisk is a drug delivery system that can automatically reset and adhere to the gastric mucosa according to the design of a leopard turtle turning over. The internal spring can insert the insulin needle into the gastric wall, but will not pierce the outer layer of the gastric wall. The team conducted in vivo studies of SOMA in rats and pigs using insulin as a model drug, and the blood drug levels were comparable to subcutaneous injection. The RaniPill developed by Rani Therapeutics consists of compartments filled with citric acid and sodium bicarbonate. When it enters the intestines, the compartment barrier is eroded, and after mixing, CO2 expands to push the dissolvable sugar-based microneedle through the outer layer of the capsule to penetrate the epithelium. RaniPill can deliver more than 10 kinds of antibodies, polypeptides and protein drugs. In large animal experiments, oral administration of 3 mg of insulin is equivalent to subcutaneous injection of 80 units, and the bioavailability is more than 50%. In the completed phase I clinical study of placebo delivery, it showed good safety. The luminal unfolding microneedle injector (LUMI) is another oral microneedle developed by the SOMA development team. The system consists of an enteric capsule, a spring and three degradable unfolding arms. When the pH is greater than or equal to 5.5, the surface coating of the capsule dissolves, and the compressed spring pushes the LUMI out of the capsule and unfolds, ensuring that the microneedle always adheres to the intestinal wall. By optimizing the unfolding force, this system has no risk of perforation in the isolated intestines of humans and pigs, and the oral bioavailability of pigs is more than 10% relative to subcutaneous injection. LUMI can also deliver vaccines, monoclonal antibodies, hormones and RNA and other macromolecular drugs that have oral barriers.

[0010] In summary, the existing technology has limitedly solved the difficult problems of oral delivery of polypeptide drugs, and a few oral polypeptide products have successfully developed and marketed. However, there are still problems in the scope of application, safety, absorption effect and stability, which need to be further solved and optimized.

[0011] Acylcarnitines are compounds generated by the combination of fatty acids and carnitines when fatty acids are transported to the inner membrane of mitochondria in vivo, and are generated from acyl-coenzyme A and carnitines due to the action of carnitine palmitoyltransferase I present in the outer membrane of mitochondria. It has been reported that acylcarnitines can be used as absorption enhancers to promote the absorption of drugs such as polypeptides at the cellular level (Anwer, W., Ratto Velasquez, A., & Tsoukanova, V. (2020). Acylcarnitines at the Membrane Surface: Insertion Parameters for a Mitochondrial Leaflet Model. Biophysical journal, 118(5), 1032-1043.). The Peptelligence® technology developed by Enteris Corporation uses citric acid as a pH adjuster, acylcarnitines as a penetration enhancer, and enteric delivery of polypeptides. There are dozens of products developed using this technology, including Ovarest, Tobrate, TbriaTM, etc. In addition, existing related technologies have proposed the use of acylcarnitines as absorption enhancers for the development of oral formulations of polypeptide molecules. However, as shown in the above description, the existing technology still has room for improvement and optimization in terms of stability, safety, and absorption effect improvement. SUMMARY

[0012] In a first aspect, the present application provides an oral polypeptide composition, which comprises a polypeptide molecule (A), a surfactant (B), and an amino acid single molecule or double molecule combination (C).

[0013] Further, the components of the oral polypeptide composition can interact to form nanoparticles, and the diameter of the nanoparticles is 0-1000 nm.

[0014] Further, the oral polypeptide composition further comprises a pH adjuster (D).

[0015] Further, the action site of the oral polypeptide composition is the small intestine and / or large intestine, and the composition can be stably absorbed in the small intestine and / or large intestine.

[0016] Further, the polypeptide molecule is a compound comprising a plurality of amino acids or containing at least one peptide bond and pharmaceutically acceptable salts thereof, and the molecular weight of the polypeptide molecule is 0.1 kDa-20 kDa, preferably 0.1 kDa-15 kDa, 0.1 kDa-10 kDa, 0.1 kDa-9.0 kDa, 0.1 kDa-5.0 kDa, 0.1 kDa-2.0 kDa; 0.5 kDa-20 kDa, 0.5 kDa-15 kDa, 0.5 kDa-10 kDa, 0.5 kDa-9.0 kDa, 0.5 kDa-5.0 kDa, 0.5 kDa-2.0 kDa; 1.0 kDa-20 kDa, 1.0 kDa-15 kDa, 1.0 kDa-10 kDa, 1.0 kDa-9.0 kDa, 1.0 kDa-5.0 kDa, 1.0 kDa-2.0 kDa; 5.0 kDa-20 kDa, 5.0 kDa-15 kDa, 5.0 kDa-10 kDa, 5.0 kDa-9.0 kDa; 10.0 kDa-20 kDa, 10.0 kDa-15 kDa;

[0017] Further, the polypeptide molecule is a compound comprising a plurality of amino acids or containing at least one peptide bond and pharmaceutically acceptable salts thereof, and the molecular weight of the polypeptide molecule is 0.1 kDa-20 kDa, preferably 0.1 kDa-15 kDa, 0.1 kDa-10 kDa, 0.1 kDa-9.0 kDa, 0.1 kDa-5.0 kDa, 0.1 kDa-2.0 kDa; 0.5 kDa-20 kDa, 0.5 kDa-15 kDa, 0.5 kDa-10 kDa, 0.5 kDa-9.0 kDa, 0.5 kDa-5.0 kDa, 0.5 kDa-2.0 kDa; 1.0 kDa-20 kDa, 1.0 kDa-15 kDa, 1.0 kDa-10 kDa, 1.0 kDa-9.0 kDa, 1.0 kDa-5.0 kDa, 1.0 kDa-2.0 kDa; 5.0 kDa-20 kDa, 5.0 kDa-15 kDa, 5.0 kDa-10 kDa, 5.0 kDa-9.0 kDa; 10.0 kDa-20 kDa, 10.0 kDa-15 kDa;

[0018] Further, the polypeptide molecule includes linear structure and cyclic structure.

[0019] Further, the polypeptide molecule includes modified peptides, derivatized peptides and peptidomimetics.

[0020] Further, the polypeptide molecules include, but are not limited to, glucagon-like peptide- 1 (GLP-1), GLP-1 analogs, GLP-1 agonists, semaglutide, liraglutide, exenatide, exenatide-4, lixisenatide, taspoglutide, langlenatide, GLP-1 (7-37), GLP-1 (7-36)NH2, dual agonists of GLP-1 receptor, glucagon receptor, oxyntomodulin, GLP-2, GLP-2 agonists or analogs, goserelin, buserelin, peptide YY (PYY), PYY analogs, glatiramer, leuprolide, deamidated vasopressin, glycopeptide antibiotics, bortezomib, corticotropin, somatorelin, luteinizing hormone-releasing hormone, calcitonin, pentagastrin, oxytocin, nesiride, enfuvirtide, eptifibatide, cyclosporine, glucagon, violamycin, thyrotropin-releasing hormone (TRH), leucine- enkephalin, methionine-enkephalin, substance P (CAS No. 33507-63-0), a parathyroid hormone (PTH) fragment, linaclotide, carfilzomib, atebant, cilengitide, and prostaglandin F2a receptor modulators, and pharmaceutically acceptable salts thereof.

[0021] In some embodiments, the GLP-1 analog is selected from acylated GLP-1 analogs, diacylated GLP-1 analogs, long-acting albumin-bound fatty acid derivatized GLP-1 analogs.

[0022] In some embodiments, the GLP-2 agonists or analogs include, but are not limited to, teduglutide and elsiglutide.

[0023] In some embodiments, the somatostatin analogs include, but are not limited to, octreotide and lanreotide or pasireotide.

[0024] In some embodiments, the goserelin includes, but is not limited to, goserelin acetate.

[0025] In some embodiments, the glatiramer includes, but is not limited to, glatiramer acetate.

[0026] In some embodiments, the leuprolide includes, but is not limited to, leuprolide acetate.

[0027] In some embodiments, the deamidated vasopressin includes, but is not limited to, deamidated vasopressin acetate and deamidated vasopressin monoacetate trihydrate.

[0028] In some embodiments, the glycopeptide antibiotics include, but are not limited to, glycosylated cyclic or polycyclic non-ribosomal peptides.

[0029] Furthermore, the glycosylated cyclic or polycyclic nonribosomal peptides include, but are not limited to, vancomycin, teicoplanin, tervacin, bleomycin, ramoranine or decaplanin, bortezomib, corticotropin, sermorelin, and luteinizing hormone-releasing hormone.

[0030] In some embodiments, the calcitonin includes, but is not limited to, salmon calcitonin.

[0031] In some embodiments, the α-parathyroid hormone (PTH) fragments include, but are not limited to, teriparatide, PTH(1-31), and PTH(2-34).

[0032] In some embodiments, the polypeptide molecule is a peptide drug, and the prostaglandin F2a receptor modulator is selected from PDC31.

[0033] Furthermore, the surfactant may be an acylcarnitine compound and / or have a carbon chain length of C8 to C90. 12 Alkyl glycosides and their pharmaceutically acceptable salts or solvates; the structural formula of the acylcarnitine compound is shown in Formula I:

[0034]

[0035] Formula I;

[0036] In formula I; R 1 It can be a carbon chain length of C6 to C6. 14 Alkyl compounds.

[0037] Furthermore, the surfactants include, but are not limited to, myristoyl-L-carnitine, decanoyl-L-carnitine, lauroyl-L-carnitine, dodecyl-beta-D-maltodextrin, tetradecyl-β-D-maltodextrin, and β-dodecyl-D-glucopyranoside.

[0038] Furthermore, the amino acid single molecule is selected from any one or more of glycine, alanine, valine, leucine, proline, tryptophan, serine, cysteine, phenylalanine, asparagine, glutamine, threonine, aspartic acid, glutamic acid, lysine, arginine, and citrulline.

[0039] Furthermore, the amino acid bimolecule is selected from any two amino acid monomolecules selected from glycine, alanine, valine, leucine, proline, tryptophan, serine, cysteine, phenylalanine, asparagine, glutamine, threonine, aspartic acid, glutamic acid, lysine, arginine, and citrulline linked by peptide bonds.

[0040] Further, the amino acid dimolecule can be selected from one or more of diglycine, di-alanine, di-valine, di-leucine, di-proline, di-tryptophan, di-serine, di-cysteine, di-phenylalanine, di-asparagine, di-glutamine, di-threonine, di-aspartic acid, di-glutamic acid, di-lysine, di-arginine and di-citrulline.

[0041] Further, the pH adjusting agent of the D component includes, but is not limited to, tartaric acid, oxalic acid, malic acid, citric acid, vitamin C, acetic acid, succinic acid, oxalic acid and succinic acid, and their hydrates and other chemical structures stable physical forms.

[0042] Further, the amount of the pH adjusting agent in the composition ranges from 1 to 150 mg, 5 to 145 mg, 10 to 140 mg, 15 to 130 mg, 20 to 120 mg, 25 to 110 mg, 20 to 100 mg, 30 to 90 mg, 40 to 80 mg.

[0043] Further, the mass ratio of the polypeptide molecule (A), the surfactant (B), and the amino acid monomolecule or dimolecule combination (C) in the composition is 0 to 5:0.1 to 20:0.5 to 20, preferably 2 to 5:5 to 20:3 to 20, further preferably 1:6:10; 1:20:3; 1:10:20; 1:15:7.5; 1:8:4; 5:20:15.

[0044] In one embodiment, the polypeptide composition is semaglutide, lauroyl-L- carnitine and diglycine; the mass ratio of the composition is 1:6:10.

[0045] In one embodiment, the composition is semaglutide, L-octanoyl carnitine and arginine; the mass ratio of the composition is 1:6:10.

[0046] In one embodiment, the composition is octreotide and L-octanoyl carnitine (10); the mass ratio of the composition is 1:10.

[0047] In one embodiment, the composition is thymopentin, diglycine and citric acid; the mass ratio of the composition is 1:20:3.

[0048] In one embodiment, the composition is thymopentin, lauroyl-L-carnitine, diglycine and citric acid; the mass ratio of the composition is 1:10:20:3.

[0049] In one embodiment, the composition is octreotide, L-octanoyl carnitine and diglycine; the mass ratio of the composition is 1:15:7.5.

[0050] In one embodiment, the composition is linaclotide, L-octanoyl carnitine, and diglycine; the mass ratio of the composition is 1 : 15 : 7.5.

[0051] In one embodiment, the composition is semaglutide and lauroyl-L-carnitine; the mass ratio of the composition is 1 : 0.1.

[0052] In one embodiment, the composition is semaglutide and lauroyl-L-carnitine; the mass ratio of the composition is 1 : 0.2.

[0053] In one embodiment, the composition is semaglutide and lauroyl-L-carnitine; the mass ratio of the composition is 1 : 0.3.

[0054] In one embodiment, the composition is semaglutide and lauroyl-L-carnitine; the mass ratio of the composition is 1 : 0.4.

[0055] In one embodiment, the composition is semaglutide and lauroyl-L-carnitine; the mass ratio of the composition is 1 : 0.6.

[0056] In one embodiment, the composition is semaglutide, lauroyl-L-carnitine, diglycine, and citric acid; the mass ratio of the composition is 1 : 8 : 4 : 4.

[0057] In one embodiment, the composition is semaglutide, lauroyl-L-carnitine, arginine, and citric acid; the mass ratio of the composition is 1 : 8 : 4 : 4.

[0058] In one embodiment, the composition is semaglutide, lauroyl-L-carnitine, diglycine, arginine, and citric acid; the mass ratio of the composition is 1 : 8 : 4 : 4 : 4.

[0059] In one embodiment, the composition is semaglutide, lauroyl-L-carnitine, and arginine; the mass ratio of the composition is 1 : 8 : 4.

[0060] In one embodiment, the composition is semaglutide, lauroyl-L-carnitine, diglycine, and citric acid; the mass ratio of the composition is 5 : 20 : 15 : 20.

[0061] In one embodiment, the composition is semaglutide, lauroyl-L-carnitine, diglycine, and citric acid; the mass ratio of the composition is 5 : 20 : 15 : 10.

[0062] In one embodiment, the composition is semaglutide, L-octanoyl carnitine, diglycine, and citric acid; the mass ratio of the composition is 5 : 20 : 15 : 20.

[0063] In a second aspect, the present application provides a pharmaceutical preparation, which comprises a combination of a polypeptide molecule (A), a surfactant (B) and a combination of amino acid monomers or dimers (C), and a pharmaceutically acceptable carrier.

[0064] Further, the polypeptide molecule (A), the surfactant (B) and the combination of amino acid monomers or dimers (C) are consistent with the first aspect of the present application.

[0065] Further, the oral preparation includes tablets, capsules, capsules within capsules, micro-patch systems within capsules, lozenges, tablets, Ovules, solutions, emulsions, suspensions, syrups, elixirs, powders and granules for reconstitution, dispersible powders and granules, pharmaceutically acceptable gums, chewable tablets, effervescent tablets and multi-particulate dosage forms.

[0066] Further, the pharmaceutically acceptable carrier can include fillers, glidants, excipients, granulation binders, lubricants, disintegrants and the like.

[0067] Further, the fillers include, but are not limited to, starches, sugars, celluloses and inorganic salts.

[0068] Further, the excipients include, but are not limited to, non-reducing sugars, microcrystalline cellulose, sodium citrate, calcium carbonate, calcium hydrogen phosphate and glycine, disintegrants such as starch (preferably corn, potato or tapioca starch), sodium starch glycolate, cross-linked sodium carboxymethyl cellulose and certain complex silicates.

[0069] Further, the granulation binders include, but are not limited to, polyvinylpyrrolidone, hydroxypropyl methylcellulose (HPMC), hydroxypropyl cellulose (HPC), sucrose and gum arabic.

[0070] Further, the lubricants include, but are not limited to, magnesium stearate, stearic acid, glyceryl behenate and talc. Similar types of solid compositions can also be used as fillers in hard capsules.

[0071] Further, the pharmaceutically acceptable carrier further includes various sweeteners, flavoring agents, colorants or dyes, combinations with emulsifying and / or suspending agents and combinations with diluents such as water, ethanol, propylene glycol and glycerol, and combinations thereof.

[0072] In a third aspect, a use of a composition in the manufacture of a medicament for treating a disease; the composition comprises a combination of a polypeptide molecule (A), a surfactant (B) and a combination of amino acid monomers or dimers (C), and a pharmaceutically acceptable carrier.

[0073] Further, the diseases include, but are not limited to, endocrine diseases including glucose metabolism disorders, diabetes, obesity, hormone deficiency and osteoporosis; neurodegenerative diseases including Alzheimer's disease and other forms of dementia, Parkinson's disease, multiple sclerosis and Huntington's disease; cardiovascular diseases including atherosclerosis, hyper-coagulable state and hypocoagulable state, coronary artery disease and cerebrovascular events; hypertension, metabolic disorders including obesity and vitamin deficiency; kidney diseases including renal failure; blood diseases including different entities of anemia; immunological and rheumatic diseases including autoimmune diseases and immunodeficiency; inflammatory diseases, infectious diseases including viral, bacterial, fungal and parasitic infections; neoplastic diseases; and multifactorial diseases including chronic pain, depression, different fibrotic states and short stature.

[0074] Advantages

[0075] The composition system of the present application significantly improves the stability of polypeptide drugs in small intestinal fluid and colon fluid; at the same time, in vivo, the composition system significantly promotes the absorption of polypeptide drugs. BRIEF DESCRIPTION OF DRAWINGS

[0076] Figure 1 Stability analysis of semaglutide and its combination system in the small intestine.

[0077] Figure 2 Stability analysis of octreotide and its combination system in the small intestine.

[0078] Figure 3 Stability analysis of thymopentin and its combination system in the small intestine.

[0079] Figure 4 Stability analysis of octreotide and its combination system in the colon fluid.

[0080] Figure 5 Stability analysis of linaclotide and its combination system in the colon fluid.

[0081] Figure 6 OLP-C102 TEM characterization.

[0082] Figure 7 OLP-C103 TEM characterization.

[0083] Figure 8 OLP-C104 TEM characterization.

[0084] Figure 9 OLP-C105 TEM characterization.

[0085] Figure 10 Rat intestinal absorption analysis of the composition system. The effect is based on the blood concentration of polypeptide drugs, the higher the concentration, the better the effect.

[0086] Figure 11 Analysis of the absorption effect of single / double amino acid components in the composition system on semaglutide.

[0087] Figure 12 Analysis of the absorption promoting effect of pH regulators in the composition system on semaglutide.

[0088] Figure 13 In vivo absorption analysis in rats.

[0089] Figure 14 In vivo absorption analysis in beagle dogs. DETAILED DESCRIPTION

[0090] The specific embodiments of the present application are described below. It should be noted that the description of these embodiments is intended to help understand the present application, but does not constitute a limitation on the present application. In addition, the technical features involved in the following described embodiments can be combined with each other as long as they do not conflict with each other.

[0091] In this document, the molecular weight of "polypeptide molecules", "peptides", "proteins", "protein drugs", "peptide molecule compounds" and "peptide molecule drugs" is specified in units of Daltons (Da), which is a unified alternative name for atomic mass units (u). The term "kDa" means 1000 Da.

[0092] The terms "polypeptide molecules", "peptides", "peptide molecule compounds" and "peptide molecule drugs", "polypeptides" and "proteins" described herein are used interchangeably herein to refer to polymers of amino acid residues, including amino acid chains of any length, including full-length proteins, in which the amino acid residues are connected by covalent peptide bonds, with a molecular weight range of about 0.1 kDa-20 kDa; also including 0.1 kDa, 0.5 kDa, 1 kDa, 1.5 kDa, 2 kDa, 2.5 kDa, 3 kDa, 3.5 kDa, 4 kDa, 4.5 kDa, 5 kDa, 5.5 kDa, 6 kDa, 6.5 kDa, 7 kDa, 7.5 kDa, 8 kDa, 8.5 kDa, 9 kDa, 9.5 kDa, 10 kDa, 15 kDa, 20 kDa.

[0093] The term "peptidomimetic" described herein refers to a small protein-like chain designed to mimic a peptide.

[0094] The peptide drug substance of the present application can be any peptide suitable for use as a pharmaceutical agent; for example, the peptide drug substance can be a linear peptide drug substance or a cyclic peptide drug substance (e.g., a cyclic peptide drug substance cyclized via at least one amide bond). It can also be a modified or derivatized peptide drug substance, such as a pegylated peptide drug substance or a fatty acid acylated peptide drug substance or a fatty diacid acylated peptide drug substance, or it can be an unmodified peptide drug substance. In particular, at its N-terminus and / or at its C-terminus, it can be unmodified, i.e., it can have a free N-terminus (-NH2) and / or a free C-terminus (-COOH) so that the drug substance can have a free (unmodified) N-terminus, or it can have a free (unmodified) C-terminus, or it can have a free N-terminus and a free C-terminus. Furthermore, the peptide drug substance can be free of histidine residues and / or free of cysteine residues.

[0095] In some embodiments, the polypeptide molecule comprises a protein, such as, but not limited to, a therapeutic agent, a nutraceutical, a glycosaminoglycan, a lipid, a carbohydrate, a steroid, a hormone, a growth hormone (GH), a growth hormone releasing hormone (GHRH), an epithelial growth factor, a vascular endothelial growth and permeability factor (VEGPF), a nerve growth factor, a cytokine, an interleukin, an interferon, a GMCSF, a hormonal product, a neurotrophic factor, a neurotrophin, a neurotransmitter, a neuromodulator, an enzyme, an antibody, a peptide, a protein fragment, a vaccine, an adjuvant, an antigen, an immunostimulatory or inhibitory factor, a hematopoietic factor, an anti-cancer product, an anti-inflammatory agent, an antiparasitic compound, an antimicrobial agent, a nucleic acid fragment, a plasmid DNA vector, a cell proliferation inhibitor or activator, a cell differentiation factor, a blood clotting factor, an immunoglobulin, an anti-angiogenic product, a negative selection marker, a toxic compound, an anti-angiogenic agent, a polypeptide, and an anticancer agent, a nucleotide, and the like, and structural analogs thereof.

[0096] In some embodiments, the polypeptide molecule includes, but is not limited to, glucagon-like peptide-1 (GLP-1), GLP-1 analogs, GLP-1 agonists (also referred to as "glucagon-like peptide-1 receptor agonists" or "GLP-1 receptor agonists"), semaglutide, liraglutide, exenatide, exenatide-4, lixisenatide, taspoglutide, langlenatide, GLP-1 (7-37), GLP-1 (7-36)NH2, dual agonists of GLP-1 receptor and glucagon receptor, oxyntomodulin, GLP-2, GLP-2 agonists or analogs, goserelin, buserelin, peptide YY (PYY), PYY analogs, glatiramer, leuprolide, deamidated vasopressin, glycopeptide antibiotics, bortezomib, corticotropin, somatorelin, luteinizing hormone-releasing hormone, calcitonin, pentagastrin, oxytocin, nesiride, enfuvirtide, eptifibatide, cyclosporine, glucagon, violamycin, thyrotropin-releasing hormone (TRH), leucine- enkephalin, methionine-enkephalin, substance P (CAS No. 33507-63-0), a parathyroid hormone (PTH) fragment, linaclotide, carfilzomib, ibatixant, cilengitide, and prostaglandin F2a receptor modulators, and pharmaceutically acceptable salts thereof.

[0097] In some embodiments, the GLP-1 analog is selected from acylated GLP-1 analogs, diacylated GLP-1 analogs, long-acting albumin-bound fatty acid derivatized GLP-1 analogs.

[0098] In some embodiments, the GLP-2 agonist or analog includes, but is not limited to, teduglutide and elsiglutide.

[0099] In some embodiments, the somatostatin analog includes, but is not limited to, octreotide and lanreotide or pasireotide.

[0100] In some embodiments, the goserelin includes, but is not limited to, goserelin acetate.

[0101] In some embodiments, the glatiramer includes, but is not limited to, glatiramer acetate.

[0102] In some embodiments, the leuprolide includes, but is not limited to, leuprolide acetate.

[0103] In some embodiments, the deamidated vasopressin includes, but is not limited to, deamidated vasopressin acetate and deamidated vasopressin monoacetate trihydrate.

[0104] In some embodiments, the glycopeptide antibiotic includes, but is not limited to, glycosylated cyclic or polycyclic non-ribosomal peptides.

[0105] Further, the glycosylated cyclic or polycyclic non-ribosomal peptide includes, but is not limited to, vancomycin, teicoplanin, telavancin, bleomycin, ramoplanin or decaplanin), bortezomib, corticotropin, sermorelin, luteinizing hormone-releasing hormone (LHRH; also known as "gonadorelin").

[0106] In some embodiments, the calcitonin includes, but is not limited to, salmon calcitonin.

[0107] In some embodiments, the alpha parathyroid hormone (PTH) fragment includes, but is not limited to, teriparatide (also known as "PTH (1-34)"), PTH (1-31), and PTH (2-34).

[0108] In some embodiments, the prostaglandin F2a receptor modulator is selected from PDC31.

[0109] The term "non-natural amino acid" as used herein refers to an amino acid that is not one of the 20 common amino acids (i.e., alanine, cysteine, aspartic acid, glutamic acid, phenylalanine, lysine, histidine, isoleucine, lysine, leucine, methionine, asparagine, proline, glutamine, arginine, serine, threonine, valine, tryptophan, and tyrosine), or pyrolysine or selenocysteine. Other terms that can be used synonymously with the term "non-natural amino acid" are "non-naturally encoded amino acid," "unnatural amino acid," and "non-naturally occurring amino acid." The term "non-natural amino acid" includes, but is not limited to, amino acids that are naturally occurring through modification of naturally encoded amino acids, including, but not limited to, the 20 common amino acids or pyrolysine and selenocysteine, but are not incorporated into growing polypeptide chains by the translation machinery itself. Examples of naturally occurring amino acids that are not naturally encoded include, but are not limited to, N-acetylglucosaminyl-L-serine, N-acetylglucosaminyl-L-threonine, and O-phosphotyrosine. In addition, the term "non-natural amino acid" includes, but is not limited to, amino acids that are not naturally occurring and can be obtained synthetically, or can be obtained by modification of a non-natural amino acid. Non-natural amino acids can include amino acids containing D-isomer configurations. In addition to natural amino acids, the amino acids can be D-amino acids or non-natural amino acids, and the molecular structure can further comprise other substituents or modifications. For example, if the peptide active ingredient is salmon calcitonin, the salmon calcitonin can be amidated at its C-terminus. Some peptides can be amidated at positions that are not naturally amidated, or can be otherwise modified.

[0110] The term "salt" in the term "pharmaceutically acceptable salt" as described herein is meant to include both water- and oil-soluble or dispersible salts, such as acetate, amsonate (4,4-diaminostilbene-2,2'-disulfonate), benzenesulfonate, benzoate, bicarbonate, bisulfate, bitartrate, borate, bromide, butyrate, calcium edetate, camsylate, carbonate, chloride, citrate, clavulariate, dihydrochloride, edetate, edisylate, estolate, esylate, fumarate, gluceptate, gluconate, glutamate, glycollylarsanilate, hexafluorophosphate, hexylresorcinate, hydrabamine, hydrobromide, hydrochloride, hydroxynaphthoate, iodide, isothionate, lactate, lactobionate, laurate, malate, maleate, mandelate, mesylate, methylbromide, methylnitrate, methylsulfate, mucate, napsylate, nitrate, N-methylglucamine ammonium salt, 3-hydroxy-2-naphthoate, oleate, oxalate, palmitate, pamoate (1,1-methene-bis-2-hydroxy-3-naphthoate, embonate), pantothenate, phosphate / diphosphate, picrate, polygalacturonate, propionate, p-toluenesulfonate, salicylate, stearate, subacetate, succinate, sulfate, sulfosaliculate, suramate, tannate, tartrate, teoclate, tosylate, triethiodide, and valerate.

[0111] The compositions of the present application can additionally comprise a pharmaceutically acceptable carrier, which is an aqueous or non-aqueous agent, e.g., alcoholic or oily, or mixtures thereof, and can contain surface active agents, emollients, lubricants, stabilizers, dyes, fragrances, preservatives, acids or bases for adjusting pH, solvents, emulsifiers, gelling agents, emollients, stabilizers, humectants, time release agents, wetting agents, or other ingredients normally included in pharmaceutical compositions in special form. Pharmaceutically acceptable carriers are known in the art and include, for example, aqueous solutions such as water or physiologically buffered saline or other solvents or carriers such as glycols, glycerol, and oils such as olive oil or injectable organic esters. The pharmaceutically acceptable carrier can contain physiologically acceptable compounds, e.g., compounds that act, e.g., to stabilize or increase the absorption of a particular inhibitor, such as carbohydrates, e.g., glucose, sucrose or dextrans, antioxidants, e.g., ascorbic acid or glutathione, chelating agents, low molecular weight proteins or other stabilizers or excipients. The pharmaceutically acceptable carrier can also be selected from substances such as distilled water, benzyl alcohol, lactose, starch, talc, magnesium stearate, polyvinylpyrrolidone, alginic acid, silicone, titanium dioxide, and flavoring agents.

[0112] The composition formulations described herein are intended to provide a composition system that can be used in an orally administrable dosage form, the form of which includes, for example, tablets (e.g., coated or uncoated tablets), capsules (e.g., gelatin capsules or HPM capsules), capsules within capsules, micro-patch systems within capsules, lozenges, troches, Ovules, solutions, emulsions, suspensions, syrups, elixirs, powders and granules for reconstitution, dispersible powders and granules, pharmaceutically acceptable gums, chewable tablets, effervescent tablets, and multiple particulate dosage forms.

[0113] The composition formulations described herein tablets, pills, capsules, and the like, can also contain binders such as acacia, arabic gum, corn starch, or gelatin; excipients such as dicalcium phosphate; disintegrating agents such as corn starch, potato starch, or alginic acid; lubricants such as magnesium stearate; and sweetening agents such as sucrose, lactose, or saccharin when a dosage unit form is a capsule, it can contain a liquid carrier such as fatty oil.

[0114] The diseases, conditions, or disorders contemplated by the present application include, but are not limited to, endocrine disorders, including disorders of glucose metabolism, diabetes, obesity, hormone deficiencies, and osteoporosis; neurodegenerative diseases, including Alzheimer's disease and other forms of dementia, Parkinson's disease, multiple sclerosis, and Huntington's disease; cardiovascular diseases, including atherosclerosis, hyper- and hypo-coagulable states, coronary artery disease, and cerebrovascular events; hypertension, metabolic disorders, including obesity and vitamin deficiencies; renal diseases, including renal failure; hematological diseases, including anemias of different entities; immunological and rheumatic disorders, including autoimmune diseases and immunodeficiencies; inflammatory diseases, infectious diseases, including viral, bacterial, fungal, and parasitic infections; neoplastic diseases; and multifactorial diseases, including chronic pain, depression, different fibrotic states, and short stature.

[0115] The term "subject" or "patient" as used herein can be an animal (e.g., a non-human animal), a vertebrate, a mammal, a rodent (e.g., a guinea pig, a hamster, a rat, a mouse), a murine animal (e.g., a mouse), a canine animal (e.g., a dog), a feline animal (e.g., a cat), a porcine animal (e.g., a pig), an equine animal (e.g., a horse), a primate, a simian animal (e.g., a monkey or an ape), a monkey (e.g., a marmoset, a baboon), or a human. It is also contemplated in the context of the present application that economically or agriculturally important animals are treated. Non-limiting examples of agronomically important animals are sheep, cattle, and pigs, while, for example, cats and dogs can be considered as economically important animals. Preferably, the subject / patient is a mammal; more preferably, the subject / patient is a human or a non-human mammal (e.g., a guinea pig, a hamster, a rat, a mouse, a rabbit, a dog, a cat, a horse, a monkey, an ape, a marmoset, a baboon, a gorilla, a chimpanzee, an orangutan, a gibbon, a sheep, a cow, or a pig).

[0116] Octreotide was first synthesized in 1979 and is an octapeptide that mimics the pharmacology of the natural somatostatin hormone, although it is a more potent inhibitor of growth hormone, glucagon, and insulin than the natural hormone. Octreotide or other somatostatin analogs can be administered in accordance with one or more embodiments of the present application for the treatment or prevention of a subject suffering from acromegaly, abnormal gastrointestinal motility, flushing episodes associated with carcinoid syndrome, portal hypertension, endocrine tumors (e.g., benign tumors, vasoactive intestinal peptide tumors), gastroparesis, diarrhea, pancreatic fistula, or pancreatic pseudocyst.

[0117] The experimental methods used in the following examples are conventional unless otherwise specified, and the experimental materials used in the following examples are commercially available unless otherwise specified.

[0118] Example 1 Stability Test

[0119] 1.1 Small Intestine Stability Test

[0120] Preparation of small intestine fluid: The small intestine was isolated directly from the animal, and the contents of the small intestine were squeezed out and collected.

[0121] Small intestine stability: The composition system was mixed with the small intestine fluid at a ratio of 1:10, and was shaken at 100 rpm in a constant temperature shaker at 37°C. Samples were taken at 0, 0.17, 0.33, 0.5, 1, 1.5, and 2 h. The mixed samples were centrifuged at 10,000 rpm at 4°C for 10 min. The supernatant was collected for quantitative analysis of the polypeptide drug.

[0122] 1.1.1 The composition system is configured using semaglutide solution with a concentration of 1 mg / ml. The content of semaglutide in all composition systems is the same. The numbers in parentheses are the mass percentage of each component in the system.

[0123] Table 1 List of semaglutide solution composition systems in small intestine

[0124]

[0125] From the above table, it can be seen that the composition systems OLP-A101 and OLP-A102 significantly improve the stability of semaglutide in small intestine fluid. Figure 1

[0126] 1.1.2 The composition system is configured using octreotide solution with a concentration of 1 mg / ml. The content of octreotide in all composition systems is the same. The numbers in parentheses are the mass percentage of each component in the system.

[0127] Table 2 List of octreotide solution composition systems in small intestine

[0128]

[0129] The above table results show that the composition system OLP-A201 significantly improves the stability of octreotide in small intestine fluid.

[0130] 1.1.3 The composition system is configured using thymopentin solution with a concentration of 1 mg / ml. The content of thymopentin in all composition systems is the same. The numbers in parentheses are the mass percentage of each component in the system.

[0131] Table 3 List of thymopentin solution composition systems in small intestine

[0132]

[0133] From the above table, it can be seen that the composition systems OLP-A301 and OLP-A302 significantly improve the stability of thymopentin in small intestine fluid. Figure 3

[0134] 1.2 Stability experiment in colon (large intestine):

[0135] 1. Base solution configuration method:

[0136] ​​Weigh out the peptone water and yeast extract into a glass flask containing distilled water. Weigh out 0.1 g NaCl, 0.04 g K2HPO4, 0.01 g MgSO4.7H2O and 0.01 g CaCl2.6H2O into the glass flask in turn and dissolve under stirring. After all the salts are dissolved, add 0.5 g L-cysteine, 0.05 g hematin chloride, 4 mL 0.025% resazurin solution and 2 g NaHCO3. Add distilled water to make the final volume reach 1 L, continue stirring for 20-30 minutes, and wait until the solution appears red to bright red for standby.

[0137] Colonic fluid preparation method:

[0138] Weigh out 2 g of large intestine contents into a 50 mL centrifuge tube, add the base solution to 20 g. Mix with a spatula to obtain a uniform 10% colonic fluid.

[0139] Colonic stability:

[0140] Mix the composition system with 10% colonic fluid at a ratio of 1:20, and blow it. The reaction system is incubated and cultured in an anaerobic workstation (nitrogen: hydrogen: carbon dioxide = 8:1:1, ambient temperature 37°C, humidity 75%). At 0, 0.5, 1, 2, 3, 4 h, samples are taken. Mix the samples at 10000 rpm, 4°C centrifuge for 10 min. Collect the supernatant for polypeptide drug quantitative analysis.

[0141] 1.2.1 Use octreotide drug solution with a concentration of 1 mg / ml to prepare the composition system. The content of somatostatin in all composition systems is the same. The numbers in parentheses are the mass percentage of each component in the system.

[0142] Table 4 List of octreotide drug solution composition systems in the colon

[0143]

[0144] Figure 4 The results show that the composition system OLP-B101 significantly improves the stability of octreotide in human colonic fluid.

[0145] 1.2.2 Use linaclotide drug solution with a concentration of 1 mg / ml to prepare the composition system. The content of somatostatin in all composition systems is the same. The numbers in parentheses are the mass percentage of each component in the system.

[0146] Table 5 List of linaclotide drug solution composition systems in the colon

[0147]

[0148] As Figure 5As shown, the composition system OLP-B201 significantly improves the stability of linaclotide in human colon fluid.

[0149] Example 2 Characterization of composition system

[0150] The characterization of the composition system was carried out for particle size and particle size. The specific DLS (dynamic light scattering) particle size study was to add the composition system into a cuvette, and the particle size and polydispersity index (PDI) of the composition system were determined using Zetasizer Nano ZS (Malvern Instruments). The composition system was prepared using a concentration of 5 mg / ml of semaglutide drug solution, and particle size characterization and particle size study were carried out. The numbers in parentheses are the mass percentage of each component in the system. As shown in Table 6

[0151] Table 6 Particle size characterization of composition system

[0152]

[0153] In addition, TEM (transmission electron microscope) analysis was carried out for the above composition system. The specific TEM particle size study was to freeze-dry the composition system and re-dissolve it with ultrapure water. 20 μL of the re-dissolved solution was added to a copper mesh; when the liquid was basically dried, an appropriate amount of 2% phosphotungstic acid aqueous solution was added to the copper mesh and incubated for 30 s; the excess liquid on the copper mesh was absorbed with a water-absorbing paper and dried; the particle morphology of the composition system was observed by transmission electron microscope (TEM, JEM-2010, JEOL), and the TEM results are shown in Figures 6-9

[0154] In summary, the DLS and TEM data indicate that the composition system of the present application can form nanoparticles with a particle size of 0-1000 nm. This characterization structure is specifically described in the present patent and belongs to the protection scope of the composition system.

[0155] Example 3 Composition system rat intestinal absorption experiment

[0156] In this study, SD rats were used, and the intestinal opening was performed on the abdomen of the rats by experimental operation, and a communicating vessel was used to keep the opening closed in the laboratory and non-experimental state. After the rat intestinal opening operation was completed, the rats were observed for 3 days, and after recovering normal physiological activity, they were used as experimental rats for subsequent absorption experiments.

[0157] Injection experiment: On the day of the experiment, the composition system was prepared, and the intestinal injection was given according to the drug dose of 2 mg / kg. After injection, the intestinal opening was closed, and the rat orbital blood was taken at the following time points: 15 min, 30 min, 1 h, 2 h, and 4 h. The blank blood before administration was taken as the 0-point blood sample for the experiment, and the blood volume at each blood taking point was 200 ul. ​

[0158] Sample processing: The whole blood of rats was collected and centrifuged. The supernatant serum sample was taken after centrifugation and stored at -20°C. The sample was used for subsequent mass spectrometry analysis.

[0159] 2.1 The content of semaglutide in all composition systems shown in the following table is the same. The numbers in parentheses are the mass percentage of each component in the system.

[0160] Table 7 Rat intestinal absorption composition system

[0161]

[0162] As Figure 10 , after administration, the absorption effect of composition systems OLP-D101 and OLP-D102 is better than that of the blank control group (OLP-D100). The data shows that the peak blood concentration of semaglutide in all groups is reached within 1 hour.

[0163] In addition, since the single / double amino acid component has a good promoting effect on the absorption of semaglutide, it is further explored whether the use of multiple single / double amino acid components can have a synergistic effect. A new system OLP-D103 is added; compared with OLP-D101 and OLP-D102 systems, there is no synergistic effect (as Figure 11 ).

[0164] In addition, a new composition system OLP-D104 is added, and the component ratio is optimized. The amount of CA is adjusted, i.e. no CA is used, compared with OLP-D102 (containing CA). The results show that (as Figure 12 ), when no CA is used, the intestinal absorption of semaglutide is slightly reduced, but compared with the semaglutide group alone, there is still a great advantage. That is, CA is an auxiliary excipient rather than a key excipient in the system, and the use concentration of CA shows a "bell-shaped" or "linear" effect.

[0165] Example 4 Composition system oral administration experiment

[0166] First, the composition system powder is prepared according to the following scheme. Specifically, each component is precisely weighed according to the mass configuration ratio shown in the following table, and then mixed uniformly.

[0167] Table 8 Oral composition system

[0168]

[0169] 4.1 Rat experiment

[0170] The above prepared composition system powder OLP-E100, OLP-E101 and OLP-E102 were implanted into the intestinal tract of rats by surgery, and the opening was sutured, and then the rats were taken blood from the eye socket at the following time points: 30 min, 1 h, 2 h, 3 h, 4 h. The blank blood was taken as the 0-point blood sample before administration, and the blood volume at each blood collection point was 200ul. Sample processing: the rat whole blood was collected and centrifuged, and the supernatant serum blood sample was stored at-20°C after centrifugation, and then used for mass spectrometry detection analysis.

[0171] The rat absorption experiment proved that the composition system OLP-E101 and OLP-E102 showed significant effect of promoting the absorption of polypeptide drugs. Figure 13 Table 9.

[0172] Table 9 Rat in vivo absorption effect

[0173]

[0174] 4.2 Beagle dog experiment

[0175] The above prepared composition system powder OLP-E100 and OLP-E103 were given to beagle dogs by gavage, and then the beagle dogs were taken blood from the forearm vein at the following time points after administration: 30 min, 1 h, 2 h, 3 h, 4 h, 6 h, 8 h, 12 h. The blank blood was taken as the 0-point blood sample before administration, and the blood volume at each blood collection point was 1ml. Sample processing: the beagle dog whole blood was collected and centrifuged, and the supernatant serum blood sample was stored at-20°C after centrifugation, and then used for mass spectrometry detection analysis.

[0176] The beagle dog oral administration experiment proved that the composition system OLP-E103 showed a sustained and significant absorption enhancement effect of semaglutide (see Table 10). Figure 14 Table 10.

[0177] Table 10 Beagle dog in vivo absorption effect

[0178]

Claims

1. A stable absorption oral polypeptide composition comprising a polypeptide molecule (A), a surfactant (B), an amino acid mono- and / or di-molecular combination (C) and / or a pH modifier (D); the polypeptide molecule (A) is semaglutide; the surfactant (B) is lauroyl-L-carnitine and / or L-octanoyl carnitine; the amino acid mono-molecule (C) is arginine; the amino acid di-molecule (C) is diglycine; the pH modifier (D) is citric acid.

2. The oral polypeptide composition of claim 1, the components of which form nanoparticles by interaction, the diameter of the nanoparticles being selected from the range of 0-1000 nm.

3. The oral polypeptide composition of claim 1, the site of action of which is the small intestine and / or the large intestine, the composition being stably absorbed in the small intestine and / or the large intestine.

4. A stable absorption pharmaceutical preparation comprising a polypeptide molecule (A), a surfactant (B), an amino acid mono- and / or di-molecular combination (C) and / or a pH modifier (D); the polypeptide molecule (A) is semaglutide; the surfactant (B) is lauroyl-L-carnitine and / or L-octanoyl carnitine; the amino acid mono-molecule (C) is arginine; the amino acid di-molecule (C) is diglycine; the pH modifier (D) is citric acid.

5. The pharmaceutical preparation of claim 4, the site of action of which is the small intestine and / or the large intestine, the pharmaceutical preparation being stably absorbed in the small intestine and / or the large intestine.

6. The pharmaceutical preparation of claim 4, the components of which form nanoparticles by interaction, the diameter of the nanoparticles being selected from the range of 0-1000 nm.

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