Pharmaceutical composition comprising peptide derivative and albumin

Abstract

The present invention relates to technology for delivering a peptide drug and, specifically, to a pharmaceutical composition comprising albumin and a peptide comprising a fatty acid chain. Using the composition according to the present invention prevents a rapid increase in initial blood concentration compared to existing peptide drugs and minimizes blood concentration deviation, and thus can significantly reduce side effects compared to existing peptide derivatives. In addition, the dosing interval is extended, thus enhancing patient convenience.

Description

Pharmaceutical composition comprising a peptide derivative and albumin

[0001] This patent application claims priority to Republic of Korea Patent Application No. 10-2024-0076529, filed with the Korean Intellectual Property Office on June 12, 2024, the disclosure of which is incorporated herein by reference.

[0002] The present invention relates to a delivery technology for a peptide drug, and more particularly, to a pharmaceutical composition comprising a peptide including a fatty acid chain capable of controlling drug release and albumin.

[0003]

[0004] Numerous papers and patents are referenced and cited throughout this specification. The disclosures of these cited papers and patents are incorporated herein by reference in their entirety to provide a clearer understanding of the state of the art and the scope of the present invention.

[0005] Depending on the route of administration, drugs have different pharmacokinetic characteristics, especially the absorption rate and peak blood concentration (C max ) are closely related. The faster the absorption rate, the higher the peak blood concentration (C max ) to reach quickly (T max ) and is generally administered in the order of intravenous, subcutaneous or intramuscular, and oral administration. max is reduced, and T max is prolonged, and accordingly, the deviation of blood concentration (C) in the order of intravenous administration, subcutaneous or intramuscular administration, and oral administration occurs when repeated administration is administered. max Wow C min The difference between the peak trough ratio (PTR) and the peak trough ratio (PTR) decreases. The greater the deviation in blood concentration, especially C maxThe higher the Tc, the higher the therapeutic window, and the closer it is to the minimum toxic concentration (MTC), which increases the possibility of side effects. Accordingly, formulation technologies that slow the initial release of the drug and increase its duration have been developed in various ways (Ducharme et al., Pharmaceuticals 2020). This effect is a pharmacodynamic characteristic that applies equally to not only small-molecule drugs but also drugs with peptides or peptide derivatives as their main ingredients. In particular, since most peptide or peptide derivative drugs are very limited in absorption when administered orally, they are administered by subcutaneous or intravenous injection, so the initial absorption rate is very fast and the blood concentration deviation (Peak Trough Ratio) tends to be large when repeated administration. Therefore, the initial blood concentration is relatively high, which ultimately leads to adverse drug reactions, and in many cases, patients discontinue administration or reduce the dose to not achieve the desired drug effect.

[0006] Liraglutide (product name: Saxenda), a representative fatty acid derivative drug of peptides ® ), Semaglutide (brand name: Ozempic) ® , Wegovy ® ), Tirzepatide (trade name: Zepbound) ® , Mounjaro ®), insulin icodec, and insulin degludec are widely prescribed worldwide for their excellent effects on diabetes and obesity. These drugs, which are fatty acid derivatives of peptides, have very limited absorption when administered orally, so they are developed and marketed as subcutaneous injection formulations. However, side effects occur due to the rapid initial increase in blood concentration after administration, and many cases of discontinuation of administration occur due to this. Representative drugs such as liraglutide, semaglutide, and tirzepatide have a high rate of digestive side effects such as nausea and emesis, and it is known that approximately 7-25% of patients discontinue administration within 6 months. In particular, in order to reduce side effects caused by rapid increases in blood concentration during initial administration, a dose escalation period is required in which the dose is gradually increased for about 2 to 3 months, after which a constant maintenance dose is administered. However, this inconvenience also causes problems such as patients discontinuing administration, missing treatment opportunities, or increased drug costs.

[0007] These issues can become even more problematic when two peptide derivative drugs with different modes of action are developed and administered in combination or in a compounded form. This can make combination drug development or combined administration impossible, or require measures such as dose reduction. To address these issues, various technologies are being studied to minimize initial drug release, limit rapid increases in blood concentration, and extend the dosing cycle beyond the existing daily (liraglutide) or weekly (semaglutide, tirzepatide) dosing cycle.

[0008] Among them, the most actively researched drug delivery systems (DDS) include PLGA microspheres, PEG derivatization, and thermosensitive polymers. PLGA microspheres are made by forming particles of approximately 10 to 30 μm using a polymer composed of a copolymer of polylactic acid and polyglyconic acid through an appropriate method (solvent evaporation, microfluidic method, etc.), and encapsulating drugs during the particle formation process. However, this process has problems such as the initial burst effect, in which a large amount of unencapsulated drugs are initially released, making actual commercial application difficult. In particular, compared to liquid injections, the large particles increase the size of the syringe for administration, causing pain for patients, and making self-administration difficult. In addition, as microspheres decompose in the body, acidic substances such as glycolic acid and lactic acid are generated, making the area highly acidic, which poses a risk of tissue or skin necrosis (Korean Patent Publication No. 10-2022-0035361).

[0009] Drug delivery technology using polymers such as polyethylene glycol (PEG) extends the half-life in the body by covalently attaching PEGs of various molecular weights to peptides or peptide derivatives, and there are many commercially available products. However, when PEG is chemically bonded to create new substances, there is a risk of anaphylactic shock due to antibody production in the body, and concerns about reduced efficacy due to antibody production during long-term administration. Therefore, it is not suitable for patients with chronic diseases such as diabetes, obesity, and liver disease who require drug administration for several months or even years (Korean Patent Publication No. 10-2014-0083929).

[0010] In addition, technologies are being developed that mix peptides or peptide derivatives using thermosensitive polymers, and control drug release by changing viscosity depending on temperature. However, commercialization is very limited due to issues with the safety of the polymers (particularly immunogenicity), handling difficulties due to temperature-dependent viscosity changes, and difficulties in self-administration due to high viscosity (Korean Patent No. 10-1331136).

[0011] The method of conjugating peptides and albumin by covalent bonding is also commercialized as a technology to extend the half-life of the peptide itself, and a representative product is TANZEUM developed by GSK. ® (Active ingredient: Abliglutide). However, it was withdrawn from the market due to efficacy issues caused by delayed peptide release (separation from albumin), concerns about the generation of new antibodies due to albumin conjugation, and concerns about anaphylaxis.

[0012] In conclusion, there is an urgent need to develop a drug delivery technology that delays the initial drug release of peptide derivative drugs, lowers the initial absorption rate, controls rapid increases in blood concentration, extends the administration cycle, shortens the dose optimization period, and enables self-administration.

[0013]

[0014] Prior art literature

[0015] (Patent Document 01) Republic of Korea Patent Publication No. 10-2022-0035361 (Published on March 22, 2022)

[0016] (Patent Document 02) Republic of Korea Patent No. 10-1411349 (Registered on June 18, 2014)

[0017] (Patent Document 03) Republic of Korea Patent Publication No. 10-2014-0083929 (Published on July 4, 2014)

[0018] (특허문헌 04) 대한민국 등록특허 제10-1331136 (2013.11.13 등록)

[0019] (논문문헌 01) Petri 등, Exposure-response analysis for evaluation of semaglutide dose levels in type 2 diabetes,Diabetes Obes Metab. 20 (2018) 2238-2245.

[0020] (논문문헌 02) Brandon T. Milliken, Design and Evaluation of Peptide Dual-Agonists of GLP-1 and NPY2 Receptors for Glucoregulation and Weight Loss with Mitigated Nausea and Emesis, J. Med. Chem. 2021, 64, 1127-1138

[0021] (논문문헌 03) Buckley 등, Real world use of tirzepatide in the treatment of type 2 diabetes in an Arab population,Diabetes Obes Metab. 26 (2024) 3381-3391.

[0022] (논문문헌 04) Yamamoto 등, Pica in Mice as a New Model for the Study of Emesis,Methods Find Exp Clin Pharmacol, 24 (2002) 135-138.

[0023] (논문문헌 05) Kanoski 등, The role of nausea in food intake and body weight suppression by peripheral GLP-1 receptor agonists, exendin-4 and liraglutide,Neuropharmacol, 62 (2012) 1916-1927

[0024]

[0025]

[0026] The present inventors have made extensive research efforts to develop a technique that can delay the initial absorption of peptide derivative drugs during subcutaneous administration, thereby reducing side effects, decreasing blood concentration variation, shortening the dose escalation period, and extending the administration cycle. As a result, when a composition containing a peptide derivative drug and albumin is used, the maximum blood concentration (C max ) is lowered, and the time to reach peak blood concentration (T max ) was confirmed to be longer. That is, when a drug is combined with an appropriate biocompatible polymer or additive, and the drug exists in a state of being non-covalently bound to the additive, the apparent molecular size (apparent hydrodynamic diameter) becomes larger than that of the drug in a relatively unbound state (unbound or free drug), and accordingly, the time it takes to reach the blood vessels or the time it takes to be absorbed after administration can be delayed, thereby changing the pharmacodynamic characteristics of the drug.

[0027] Specifically, as a result of administering the pharmaceutical composition, the complex formed by electrostatic, non-covalent bonding of the drug and the additive had a larger apparent hydrodynamic diameter than the drug itself, and accordingly, the amount of unbound drug present in the formulation was greatly reduced, and when administered subcutaneously, release from the subcutaneous tissue was delayed, delaying drug absorption into the blood vessels. Through this, the time to reach the maximum blood concentration (T max ) is prolonged, and the peak blood concentration (C max ) was significantly lowered, and the variability of the blood concentration and trough blood concentration of the drug at steady state during repeated administration (i.e., C max / C min value, The Peak Trough Ratios (PTR) were reduced. As a result, the present invention was completed by finding that the effect of improving the efficacy and side effects can be obtained by maintaining the blood drug concentration at a constant level.

[0028] Accordingly, an object of the present invention is to provide a peptide delivery composition comprising a peptide comprising a fatty acid chain and albumin.

[0029] Another object of the present invention is to provide a method for preparing a composition for peptide delivery, comprising the step of mixing a peptide containing a fatty acid chain into a solution containing albumin.

[0030] Another object of the present invention is to provide a pharmaceutical composition for preventing or treating metabolic diseases, neurological diseases, cardiovascular diseases, endocrine diseases, digestive diseases, brain diseases, oncological diseases or combinations thereof, comprising a peptide comprising a fatty acid chain and a peptide delivery composition comprising albumin.

[0031] Other objects and advantages of the present invention will become more apparent from the detailed description, claims and drawings below.

[0032]

[0033] The present invention provides the following inventions 1 to 27.

[0034] 1. Composition for peptide delivery comprising a peptide containing a fatty acid chain and albumin:

[0035] (a) the concentration of the peptide is 0.1 mg / mL to 500 mg / mL; and

[0036] (b) The concentration of the albumin is 0.1 mg / mL to 500 mg / L.

[0037] 2. A composition for peptide delivery, wherein the weight ratio of the peptide and albumin in 1 is 1:0.6 to 1:500.

[0038] 3. A composition for peptide delivery, wherein in 1 or 2, the molar ratio of the peptide and albumin is 1:0.05 to 1:30.

[0039] 4. A composition for peptide delivery, wherein in any one of 1 to 3, the peptide and albumin are non-covalently bound to form a complex.

[0040] 5. A composition for peptide delivery, wherein in any one of 1 to 4, the unbound peptide among the peptides is 30 wt% or less of the total peptide weight.

[0041] 6. A composition for delivering a peptide, wherein in any one of 1 to 5, the albumin is composed of albumin bound to a peptide, free albumin, or a combination thereof.

[0042] 7. A composition for peptide delivery, wherein the peptide is composed of 15 to 250 amino acids in any one of 1 to 6.

[0043] 8. A composition for peptide delivery, wherein in any one of 1 to 7, the fatty acid chains are 1 to 5.

[0044] 9. A composition for peptide delivery, wherein the number of carbon atoms in the fatty acid chain included in the peptide is 8 to 30 in any one of 1 to 8.

[0045] 10. In any one of 1 to 9, the peptide is liraglutide, semaglutide, tirzepatide, dulaglutide, cotadutide, cagrilintide, cagrisema, amycretin, retatrutide, nisotirastide, pemvidutide, mazdutide, survodutide, ecnoglutide, insulin icodec, insulin degludec, somapacitan, dapiglutide, A composition for peptide delivery, wherein the composition is selected from the group consisting of petrelintide, SAR441255, NNC0165-1273, HM15136, HM15275, TE-8105, NNC0090-2746, DA-1726, Met-097i, BGM0504, VK2735 and pharmaceutically acceptable salts thereof.

[0046] 11. A composition for peptide delivery, wherein in any one of 1 to 10, the albumin is selected from the group consisting of human serum albumin (HSA), recombinant human serum albumin (rHSA), bovine serum albumin (BSA), ovalbumin (OVA), and combinations thereof.

[0047] 12. A composition for delivering a peptide, wherein the composition has the properties of reducing the peak blood concentration of the peptide, increasing the time to reach the peak blood concentration, or a combination thereof, compared to a peptide preparation alone.

[0048] 13. A composition for peptide delivery, wherein the peptide is a peptide selected from the group consisting of SEQ ID NOs: 1 to 8, in any one of 1 to 12.

[0049] 14. A composition for peptide delivery, wherein the composition comprises a complex comprising a peptide comprising a fatty acid chain and albumin, an unbound peptide, free albumin, or a combination thereof.

[0050] 15. A composition for peptide delivery, wherein the composition comprises two or more peptides comprising a fatty acid chain, in any one of 1 to 14.

[0051] 16. A method for producing a composition for peptide delivery, comprising the step of mixing a peptide containing a fatty acid chain into a solution containing albumin.

[0052] 17. A method for producing a composition for peptide delivery, wherein the weight ratio of the peptide and albumin in the composition is 1:0.8 to 1:500.

[0053] 18. A method for producing a composition for peptide delivery, wherein the concentration of the peptide and albumin in the composition is 0.1 mg / mL to 500 mg / mL in 16 or 17.

[0054] 19. A pharmaceutical composition for preventing or treating metabolic diseases, neurological diseases, cardiovascular diseases, endocrine diseases, digestive diseases, oncological diseases, or a combination thereof, comprising a peptide comprising a fatty acid chain and a peptide delivery composition comprising albumin:

[0055] The above peptide is a glucagon-like peptide-1 receptor agonist (GLP-1 RA), insulin icodec, or a combination thereof.

[0056] 20. In 19, the peptide is liraglutide, semaglutide, tirzepatide, dulaglutide, cotadutide, cagrilintide, cagrisema, amycretin, retatrutide, nisotirastide, pemvidutide, mazdutide, survodutide, ecnoglutide, insulin icodec, insulin degludec, somapacitan, dapiglutide, petrelintide, A pharmaceutical composition selected from the group consisting of SAR441255, NNC0165-1273, HM15136, HM15275, TE-8105, NNC0090-2746, DA-1726, Met-097i, BGM0504, VK2735 and pharmaceutically acceptable salts thereof.

[0057] 21. A pharmaceutical composition according to claim 19 or 20, wherein the peptide delivery composition is any one of compositions 1 to 15.

[0058] 22. A pharmaceutical composition according to any one of 19 to 21, wherein the disease is obesity, fatty liver disease, dyslipidemia, degenerative neurological disease, hypertension, heart failure, arteriosclerosis, diabetes, fatty liver disease, gastritis, inflammatory bowel disease, cancer, or a combination thereof.

[0059] 23. A pharmaceutical composition according to any one of 19 to 22, wherein the pharmaceutical composition is administered orally, subcutaneously, intramuscularly, or intravenously.

[0060] 24. A pharmaceutical composition according to any one of 19 to 23, wherein the pharmaceutical composition is administered repeatedly for one or more cycles.

[0061] 25. A pharmaceutical composition, wherein the interval between the cycles is from 1 day to 6 months, in any one of 19 to 24.

[0062] 26. A method for preventing or treating a disease, comprising administering to a subject any one of the compositions 1 to 15 or any one of the pharmaceutical compositions 19 to 25.

[0063] 27. A method for preventing or treating a disease, wherein the disease is a metabolic disease, a nervous system disease, a cardiovascular disease, an endocrine disease, a digestive system disease, a tumor disease, or a combination thereof.

[0064] 28. A method for preventing or treating a disease according to 26 or 27, wherein the disease is obesity, fatty liver disease, dyslipidemia, degenerative neurological disease, hypertension, heart failure, arteriosclerosis, diabetes, fatty liver disease, gastritis, inflammatory bowel disease, cancer, or a combination thereof.

[0065] In one aspect of the present invention, the present invention provides a peptide delivery composition comprising a peptide comprising a fatty acid chain and albumin:

[0066] (a) the concentration of the peptide is 0.1 mg / mL to 500 mg / mL; and

[0067] (b) The concentration of the albumin is 0.1 mg / mL to 500 mg / L.

[0068] The present inventors have made extensive research efforts to develop a technique that can delay the initial absorption of peptide derivative drugs during subcutaneous administration, thereby reducing side effects, decreasing blood concentration variation, shortening the dose escalation period, and extending the administration cycle. As a result, when a composition containing a peptide derivative drug and albumin is used, the maximum blood concentration (C max ) is lowered, and the time to reach peak blood concentration (T max ) was confirmed to be longer. That is, when a drug is combined with an appropriate biocompatible polymer or additive, and the drug exists in a state of being non-covalently bound to the additive, the apparent molecular size (apparent hydrodynamic diameter) becomes larger than that of the drug in a relatively unbound state (unbound or free drug), and accordingly, the time it takes to reach the blood vessels or the time it takes to be absorbed after administration can be delayed, thereby changing the pharmacodynamic characteristics of the drug.

[0069] In this specification, the peptide includes a therapeutic peptide, a variant thereof, or a derivative thereof designed to act on a specific receptor in a living body or exhibit physiological activity. The peptide derivative may be a peptide comprising a fatty acid chain. However, the present invention is not limited thereto, and includes any peptide derivative comprising a functional group capable of interacting with albumin.

[0070] In the present specification, the fatty acid chain can play a role in extending the half-life of the drug and increasing its stability in the body by enabling the peptide to bind to albumin in the body. The peptide is covalently bonded to one or more "fatty acid chains," and the fatty acid chains are generally covalently bonded to specific residues (e.g., Lys, Glu, etc.) of the peptide via a linker. These fatty acid chains provide a functional group that increases hydrophobic interactions and albumin binding affinity, and can selectively and non-covalently bind to the fatty acid binding pocket of albumin. In one embodiment of the present invention, the peptide comprising the fatty acid chain can be used in the same sense as a peptide derivative or a peptide fatty acid derivative. The peptide comprising the fatty acid chain can be provided by being artificially manufactured in vitro, or can be provided by artificially modifying or manipulating a natural peptide.

[0071] The term "peptide delivery composition" as used herein refers to a formulation designed to enable a peptide to stably reach a target site in a living body. The composition has a peak blood concentration (C) higher than that of a single formulation of the peptide or peptide derivative. max ) significantly lowered the time to peak blood concentration (T max ) can delay the absorption of the drug, thereby reducing side effects caused by rapid drug absorption. In addition, it can provide sustained drug efficacy by reducing the peak-trough ratio (PTR) of blood concentration during repeated administration, which is advantageous in extending the dosing interval or improving compliance.

[0072] Meanwhile, the composition of the present invention is distinct from "peptides covalently bound to albumin," such as fusion proteins or chemical conjugates. Conventional albumin fusion or covalent bonding methods involve the formation of genetic or chemical bonds between peptide derivatives and albumin, resulting in complex manufacturing processes, high production costs, and the potential for immunogenicity due to the modified albumin or fusion proteins. In particular, chemical conjugates have the disadvantage of being difficult to control the position or number of bonds, and low product homogeneity, making it difficult to ensure formulation consistency.

[0073] In contrast, the composition of the present invention forms a non-covalent complex by simply mixing the peptide derivative and albumin in a solution, resulting in a simple manufacturing process, low cost, and excellent structural stability and quality reproducibility. Furthermore, the presence of unbound albumin within the composition allows for more precise control of complex formation balance, drug release rate, and initial absorption delay.

[0074] This configuration is also advantageous in terms of pharmacodynamics, resulting in rapid C max Suppress the rise, T max By delaying the release of drugs, it is possible to simultaneously achieve sustained drug release and suppression of side effects, and it also has the potential to serve as a universal platform that can be easily applied to various peptide derivatives. In other words, the present invention has improved characteristics in various aspects, such as ease of manufacture, cost-effectiveness, biostability, and drug activity controllability, compared to existing covalent drugs.

[0075] The composition of the present invention can be prepared in various formulations, and the final product can be provided in a liquid or lyophilized state. In the case of a liquid formulation, the peptide derivative and albumin can be pre-mixed in an aqueous medium and present in a directly administrable state, and can be prepared as an injection, eye drop, inhalant, oral solution, etc., as needed. The lyophilized formulation can be used to ensure long-term storage stability and to reconstitute immediately before administration, and can be provided as an injectable powder (vial), a powder / solvent mixture kit, etc.

[0076] In addition, the composition of the present invention can be applied to various delivery systems such as microneedle patches, hydrogels, liposomes, nanoemulsions, nanoparticle systems, as well as water-soluble formulations, and can be administered by transdermal, mucosal, nasal, pulmonary, intravenous, intramuscular, subcutaneous, and other injection routes. Excipients, stabilizers, preservatives, buffers, etc. used in the formulation process can be selected from ingredients acceptable at a typical pharmaceutical level.

[0077] Therefore, the composition of the present invention is not limited to a specific formulation state, and can be implemented in various forms according to the purpose of maintaining efficacy, convenience of use, optimization of administration route, etc.

[0078] The concentration of the peptide or albumin in the above composition may be 0.1 mg / mL to 500 mg / mL. For example, 0.1 mg / mL to 500 mg / mL, 0.1 mg / mL to 200 mg / mL, 0.1 mg / mL to 100 mg / mL, 0.1 mg / mL to 50 mg / mL, 0.1 mg / mL to 20 mg / mL, 0.1 mg / mL to 10 mg / mL, 0.1 mg / mL to 5 mg / mL, 0.1 mg / mL to 2 mg / mL, 0.1 mg / mL to 1 mg / mL, 0.1 mg / mL to 0.5 mg / mL, 0.2 mg / mL to 500 mg / mL, 0.2 mg / mL to 200 mg / mL, 0.2 mg / mL to 100 mg / mL, 0.2 mg / mL to 50 mg / mL, 0.2 mg / mL to 20 mg / mL, 0.2 mg / mL to It may be, but is not limited to, 10 mg / mL, 0.2 mg / mL to 5 mg / mL, 0.2 mg / mL to 2 mg / mL, 0.2 mg / mL to 1 mg / mL, 0.5 mg / mL to 500 mg / mL, 0.5 mg / mL to 200 mg / mL, 0.5 mg / mL to 100 mg / mL, 0.5 mg / mL to 50 mg / mL, 0.5 mg / mL to 20 mg / mL, 0.5 mg / mL to 10 mg / mL, 0.5 mg / mL to 5 mg / mL, 0.5 mg / mL to 2 mg / mL, 0.5 mg / mL to 1 mg / mL.

[0079] The concentration of albumin in the above composition may be 0.5 mg / mL to 500 mg / mL. For example, 0.5 mg / mL to 500 mg / mL, 0.5 mg / mL to 450 mg / mL, 0.5 mg / mL to 400 mg / mL, 0.5 mg / mL to 350 mg / mL, 0.5 mg / mL to 300 mg / mL, 0.5 mg / mL to 250 mg / mL, 0.5 mg / mL to 200 mg / mL, 0.5 mg / mL to 150 mg / mL, 0.5 mg / mL to 100 mg / mL, 0.5 mg / mL to 50 mg / mL, 0.5 mg / mL to 10 mg / mL, 0.5 mg / mL to 1 mg / mL, 1 mg / mL to 500 mg / mL, 4 mg / mL to 500 mg / mL, 8 mg / mL to 500 mg / mL, 20 mg / mL to 500mg / mL, 40mg / mL to 500mg / mL, 50mg / mL to 500mg / mL, 60mg / mL to 500mg / mL, 80mg / mL to 500mg / mL, 100mg / mL to 500mg / mL, 120mg / mL to 500mg / mL, 140mg / mL to 500mg / mL, 160mg / mL to 500mg / mL, 180mg / mL to 500mg / mL, 200mg / mL to 500mg / mL, 300mg / mL to 500mg / mL, 8mg / mL to 320mg / mL, 8mg / mL to 160mg / mL, 8mg / mL to 140mg / mL, 8mg / mL to 120mg / mL, 8mg / mL to It may be, but is not limited to, 100 mg / mL, 8 mg / mL to 80 mg / mL, 8 mg / mL to 60 mg / mL, 8 mg / mL to 40 mg / mL, 8 mg / mL to 20 mg / mL, 40 mg / mL to 200 mg / mL, 60 mg / mL to 140 mg / mL, or 60 mg / mL to 100 mg / mL.

[0080] In one embodiment of the present invention, the weight ratio of the peptide derivative and albumin is 1:0.6 to 1:500.

[0081] In one embodiment of the present invention, the weight ratio of albumin to the peptide derivative of the composition is 1:0.6 to 1:500. For example, the weight ratio of the peptide derivative and albumin is 1: 0.6 to 1: 500, 1: 0.6 to 1: 450, 1: 0.6 to 1: 400, 1: 0.6 to 1: 350, 1: 0.6 to 1: 300, 1: 0.6 to 1: 250, 1: 0.6 to 1: 200, 1: 0.6 to 1: 150, 1: 0.6 to 1: 100, 1: 0.6 to 1: 80, 1: 0.6 to 1: 50, 1: 0.6 to 1: 16, 1: 0.6 to 1: 8, 1: 0.6 to 1: 1.6, 1: 0.8 to 1: 100, 1: 1 to 1: 100, 1: 1 to 1: 10, 1: 8 to 1: 100, 1: 16 to 1: 100, 1: 50 to 1: 100, 1: 80 to 1: 100, 1: 50 to 1: 500, 1: 100 to 1: 500, 1: 150 to 1: 500, 1: 200 to 1: 500, 1: 250 to 1: 500, 1: 300 to 1: 500, 1: 350 to 1: 500, 1: 400 to 1: 500, 1: 450 to 1: 500, 1: 50 to It may be, but is not limited to, 1:450, 1:100 to 1:400, 1:150 to 1:350, 1:200 to 1:300.

[0082] In one embodiment of the present invention, the molar ratio of the peptide and albumin is 1:0.05 to 1:30.

[0083] For example, the molar ratio of peptide to albumin is 1:0.05 to 1:30, 1:0.05 to 1:25, 1:0.05 to 1:20, 1:0.05 to 1:15, 1:0.05 to 1:10, 1:0.05 to 1:5, 1:0.05 to 1:1, 1:0.05 to 1:0.5, 1:0.3 to 1:30, 1:0.5 to 1:30, 1:1 to 1:30, 1:10 to 1:30, 1:20 to 1:30, 1:0.3 to 1:1, 1:0.5 to 1:1, 1:0.5 to 1:10, 1:0.5 to 1:15, 1:0.5 to It can be, but is not limited to, 1:20, 1:1 to 1:10, or 1:1 to 1:15.

[0084] In one embodiment of the present invention, the peptide and albumin are non-covalently bound to form a complex. In one specific embodiment of the present invention, the composition comprises a complex comprising a peptide comprising a fatty acid chain and albumin.

[0085] The term "complex comprising a peptide comprising a fatty acid chain and albumin" as used herein refers to a structure formed by non-covalent interactions between (i) a peptide comprising at least one fatty acid chain and (ii) albumin. Meanwhile, "albumin" does not covalently bind to the peptide within the complex, but can reversibly bind to it through hydrophobic interactions, electrostatic attraction, van der Waals forces, or ionic bonds.

[0086] This complex is generally formed in an aqueous solution, and the apparent molecular size (hydrodynamic diameter) at the polymer level is significantly increased compared to the peptide alone. When the peptide containing a fatty acid chain and albumin are mixed in an appropriate solvent for administration to a patient, the peptide and albumin electrostatically bind to form a complex, and this complex can have an apparent molecular size (apparent hydrodynamic diameter) more than 5 times that of the peptide derivative. Therefore, when administered subcutaneously, absorption into the capillaries is delayed, and the time to reach the peak blood concentration (T) is shortened. max ) and reduce the variability of blood drug concentration (i.e., the difference between the maximum and minimum concentrations, Peak Trough Ratios (PTR)) at steady state through repeated administration, thereby precisely controlling blood drug concentration, thereby improving efficacy and side effects.

[0087] The term "non-covalent bond" in this specification means a bonding form that is stabilized by physicochemical interactions such as electrostatic interaction, hydrophobic interaction, hydrogen bonding, van der Waals interaction, and ionic binding, rather than a chemical bonding through sharing of electron pairs like a covalent bond.

[0088] For example, a peptide derivative containing a fatty acid chain can bind to the fatty acid binding pockets of albumin by electrostatic or hydrophobic attraction, which is a representative example of a non-covalent bond. Since the albumin used in the present invention usually has at least 5, and in some cases at least 7, fatty acid binding pockets, a single albumin molecule can bind to multiple peptide derivatives simultaneously, and conversely, a single peptide derivative can bind to multiple albumin molecules simultaneously.

[0089] Furthermore, these complexes are not limited to a simple 1:1 combination (monocomplex), but can be formed in the form of oligomers such as dimers, trimers, tetramers, or more. This is because a single albumin molecule has multiple fatty acid binding sites, allowing it to bind to multiple peptide derivatives simultaneously. Conversely, peptide derivatives can also aggregate with each other through hydrophobic interactions, leading to mutual binding.

[0090] Because these complexes have a non-covalent macromolecular complex structure, their apparent hydrodynamic diameter can be several times larger than that of a single peptide derivative, which contributes to significantly delayed tissue absorption of the drug. Furthermore, when the complex is formed into a multimeric structure, its apparent hydrodynamic diameter is substantially larger than that of a simple 1:1 complex, and can extend to levels exceeding 100 kDa.

[0091] In addition, non-covalent complexes provide flexibility to be reversibly dissociated in vivo, thereby providing stability against metabolic enzymes in the body while maintaining a constant drug release rate, and effectively suppressing rapid increases in blood concentration that cause side effects.

[0092] In one embodiment of the present invention, the unbound peptide in the composition is 30 wt% or less of the total peptide weight. The unbound peptide may be used in the same sense as a peptide that does not form a complex with albumin. The unbound peptide may be 30 wt% or less, 25 wt% or less, 20 wt% or less, 15 wt% or less, 10 wt% or less, 5 wt% or less, 3 wt% or less, or 1 wt% or less of the total peptide weight of the composition, but is not limited thereto. The unbound peptide may be 0 to 30 wt%, 0 to 20 wt%, 0 to 10 wt%, or 0 to 5 wt% of the total peptide weight of the composition, but is not limited thereto.

[0093] In one embodiment of the present invention, the peptide is composed of 15 to 250 amino acids. For example, it may be composed of 15 to 250 amino acids, 15 to 200 amino acids, 15 to 150 amino acids, 15 to 100 amino acids, 15 to 50 amino acids, 50 to 250 amino acids, 100 to 250 amino acids, 150 to 250 amino acids, 200 to 250 amino acids, 20 to 100 amino acids, or 20 to 50 amino acids. Representative examples of the peptide are as shown in Table 1, but are not limited thereto.

[0094]

[0095] Examples of fatty acid derivative drugs of representative peptides Drug number (development name) Development company Number of amino acids Number of fatty acid chains Number of carbons in each fatty acid chain 1 Liraglutide Novo Nordisk 311162 Semaglutide Novo Nordisk 311183 Tirzepatide Eli Lilly 392204 Insulin detemir Novo Nordisk 511145 Insulin degludec Novo Nordisk 501166 Insulin icodec Novo Nordisk 501207 Somapacitan Novo Nordisk 1911188 Cagrilintide Novo Nordisk 372209 Retatrutide Eli Lilly 3922010 Survodutide BI / Zealand 2921811 Mazdutide Innovent / Lilly3911812CotadutideAstraZeneca3021813PemvidutideAltimmune2911814EcnoglutideSciwind / LG Chem3721815SAR441255Sanofi4811616DapiglutideZealand Pharma3411817PetrelintideZealand Pharma3612018NNC0165-1273Novo Nordisk3421819HM15136Hanmi Pharma2921820TE-8105 (tentative name)Immunwork312161821NNC0090-2746Novo Nordisk4011623DA-1726NeuroBo Pharmaceuticals39220

[0096] In the present invention, the peptide constituting the peptide derivative may include an amino acid sequence having about 30%, 40%, 50%, 60%, 70%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% homology with the amino acid sequence of the natural peptide. The peptide variant may include at least 5 or more, at least 10 or more, at least 15 or more, at least 20 or more, at least 30 or more, or at least 40 or more amino acids of the wild-type peptide sequence.

[0097] It should be understood that the peptides of the present invention may be provided in the form of salts or other derivatives. Salts include pharmaceutically acceptable salts, such as acid addition salts and basic salts. Examples of acid addition salts include salts of inorganic acids such as hydrochloride, sulfate, phosphate, and nitrate, and organic salts such as citrate, acetate, glycolic acid, and lactic acid. Examples of basic salts include salts in which the cation is selected from alkali metals such as sodium and potassium, alkaline earth metals such as calcium, and ammonium ion +N(R3)3(R4), wherein R3 and R4 independently represent optionally substituted C1-C6-alkyl, optionally substituted C2-C6-alkenyl, optionally substituted aryl, or optionally substituted heteroaryl. Other examples of pharmaceutically acceptable salts are described in Remington's Pharmaceutical Sciences, 17 th edition. Ed. Alfonso R. Gennaro (Ed.), Mark Publishing Company, Easton, PA, USA, 1985] and its recent revisions, and in the literature [Encyclopedia of Pharmaceutical Technology].

[0098] In one embodiment of the present invention, the composition comprises unbound albumin. The albumin included in the composition of the present invention may form a complex with a peptide, or may exist in the form of unbound albumin without forming a complex with the peptide.

[0099] The term "free albumin" as used herein refers to free albumin present in the composition, not noncovalently bound to a peptide, and in a form that does not contribute to complex formation. The presence of free albumin allows for greater control over the drug release properties of the composition and is advantageous in precisely regulating the structural stability and in vivo behavior of the complex.

[0100] In one embodiment of the present invention, the composition comprises a complex comprising a peptide comprising a fatty acid chain and albumin, an unbound peptide, free albumin, or a combination thereof.

[0101] In one embodiment of the present invention, the fatty acid chains are 1 to 5.

[0102] In one embodiment of the present invention, the number of carbon atoms in the fatty acid chain is 8 to 30. The number of carbon atoms in the fatty acid chain may be, for example, 8 to 30, 8 to 20, 8 to 10, 10 to 30, 20 to 30, 10 to 15, 15 to 20, or 10 to 20, but is not limited thereto.

[0103] In one embodiment of the present invention, the peptide derivative is liraglutide, semaglutide, tirzepatide, dulaglutide, cotadutide, cagrilintide, cagrisema, amycretin, retatrutide, nisotirastide, pemvidutide, mazdutide, survodutide, ecnoglutide, insulin icodec, insulin degludec, somapacitan, dapiglutide, petrelintide, A pharmaceutically acceptable salt thereof is selected from the group consisting of SAR441255, NNC0165-1273, HM15136, HM15275, TE-8105, NNC0090-2746, DA-1726, Met-097i, BGM0504, VK2735 and pharmaceutically acceptable salts thereof.

[0104] In one embodiment of the present invention, the peptide is a peptide selected from the group consisting of SEQ ID NOs: 1 to 8.

[0105] In one embodiment of the present invention, the albumin is selected from the group consisting of human serum albumin (HSA), recombinant human serum albumin (rHSA), bovine serum albumin (BSA), ovalbumin (OVA), and combinations thereof. However, the present invention is not limited thereto, and may include various albumins such as human-derived albumin, recombinant albumin, and animal-derived albumin. In one embodiment of the present invention, complex formation and pharmacodynamic effects with peptide derivatives were consistently confirmed for various types of albumin.

[0106] In one embodiment of the present invention, the albumin is purified from human plasma (human serum albumin), recombinantly produced from microorganisms such as yeast and E. coli, and encompasses fragments or variants of albumin having one or more biological activities of albumin. To manufacture the albumin of the present invention, an albumin solution having a certain concentration or higher may be used.

[0107] In one embodiment of the present invention, the composition has the properties of reducing the peak blood concentration of the peptide, increasing the time to reach the peak blood concentration, or a combination thereof, compared to a peptide preparation alone.

[0108] The composition comprising a peptide and albumin containing a fatty acid chain of the present invention has a maximum blood concentration (C) of a peptide-only preparation max ) lower than C max , time to reach maximum blood concentration (T max ) longer than T max It is characterized by having a low C at this time. max When the same dose was administered, the C of the peptide derivative administered alone maxIt means lower than, for example, the C of a conventional single-administered peptide derivative. max C 0.9, 0.8, 0.7, 0.6, 0.5, 0.4, 0.3, 0.2, 0.1 times or less max can have. Also, long T max When the same dose was administered, the T of the peptide administered alone max It means longer than, for example, the T of a conventional single-administered peptide derivative. max 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, 2, 3, 4, 5, 6, 7, 8, 9, 10, 20, 30, 40, 50, 60, 70, 80, 90, 100, 200, 300, 400, 500 times or more of T max can have

[0109] A composition comprising a peptide comprising a fatty acid chain of the present invention and albumin is characterized by having a blood half-life similar to or improved upon that of a peptide alone. In one embodiment, the composition may have a half-life that is improved by at least about 1.1, 1.2, 1.5, 2, 2, 3, 4, 5, 6, 7, 8, 9, 10, 20, 30, 40, 50, 60, 70, 80, 90, 100, 200, 500, 1,000, 2,000, 5,000, or 10,000 times greater than the half-life of a naturally occurring peptide.

[0110] In one embodiment of the present invention, the composition comprises two or more peptides comprising a fatty acid chain. When multiple peptides are included, the weight ratio of any first peptide to any second peptide is from 1:1 to 1:100. For example, the weight ratio of any first peptide and any second peptide may be, but is not limited to, 1:1 to 1:100, 1:1 to 1:80, 1:1 to 1:60, 1:1 to 1:40, 1:1 to 1:20, 1:1 to 1:10, 1:1 to 1:5, 1:1 to 1:3, 1:3 to 1:100, 1:5 to 1:100, 1:10 to 1:100, 1:20 to 1:100, 1:40 to 1:100, 1:60 to 1:100, 1:80 to 1:100, 1:5 to 1:20, 1:5 to 1:15, or 1:5 to 1:10.

[0111]

[0112] In one aspect of the present invention, the present invention provides a method for preparing a peptide delivery composition, comprising the step of mixing a peptide containing a fatty acid chain into a solution containing albumin.

[0113] The above mixing can be performed at room temperature (approximately 1 to 30°C), ambient temperature (approximately 15 to 25°C), or refrigerated (2 to 8°C), and a stable complex is generally formed by stirring for 30 minutes to 1 hour or more. The stirring conditions can be performed at a speed of, for example, 120 rpm, but are not limited thereto.

[0114] The solvent used can be a water-soluble solvent, preferably phosphate-buffered saline (PBS), and the pH range is preferably approximately 5.0 to 9.0. Under these conditions, the peptide derivative containing a fatty acid chain noncovalently interacts with albumin to form a complex, and the formed complex can be used directly as a pharmaceutical composition without a separate purification step. Furthermore, the present manufacturing method can be performed simply without chemical covalent bonding or complex modification steps, and thus has advantages in terms of process reproducibility and production efficiency.

[0115] In one embodiment of the present invention, the weight ratio of the peptide and albumin of the composition is 1:0.8 to 1:500. In one embodiment of the present invention, the concentration of the peptide and albumin of the composition is 0.1 mg / mL to 500 mg / mL. In one embodiment of the present invention, the molar ratio of the peptide and albumin is 1:0.05 to 1:30.

[0116]

[0117] In one aspect of the present invention, the present invention provides a pharmaceutical composition for preventing or treating metabolic diseases, neurological diseases, cardiovascular diseases, endocrine diseases, digestive diseases, brain diseases, oncological diseases or combinations thereof, comprising a peptide delivery composition comprising a complex comprising a peptide comprising a fatty acid chain and albumin:

[0118] The above peptide is a glucagon-like peptide-1 receptor agonist (GLP-1 RA), insulin icodec, or a combination thereof.

[0119] As used herein, the term "prevention" refers to the prevention or protective treatment of a disease or disease state. As used herein, the term "treatment" refers to the reduction, suppression, alleviation, or eradication of a disease state.

[0120] The pharmaceutical composition of the present invention can be manufactured in the form of a unit dose or can be manufactured by inserting it into a multi-dose container by formulating it using a pharmaceutically acceptable carrier and / or excipient according to a method that can be easily performed by a person having ordinary skill in the art to which the present invention pertains. In this case, the formulation may be in the form of a solution, suspension or emulsion in an oil or aqueous medium, or may be in the form of an extract, powder, suppository, powder, granules, tablet, capsule or injection, and may additionally include a dispersing agent or stabilizer.

[0121] The pharmaceutical composition of the present invention may include a pharmaceutically acceptable carrier. Pharmaceutically acceptable carriers are those commonly used in formulations, and include, but are not limited to, lactose, dextrose, sucrose, sorbitol, mannitol, starch, acacia gum, calcium phosphate, alginate, gelatin, calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, water, syrup, methyl cellulose, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate, and mineral oil. In addition to the above components, the pharmaceutical composition of the present invention may further include lubricants, wetting agents, sweetening agents, flavoring agents, emulsifiers, suspending agents, preservatives, and the like. Suitable pharmaceutically acceptable carriers and formulations are described in detail in Remington's Pharmaceutical Sciences (19th ed., 1995).

[0122] The pharmaceutical composition of the present invention can be administered orally or parenterally, and can be administered by, for example, intravenous injection, subcutaneous injection, intramuscular injection, intraperitoneal injection, intrasternal injection, intratumoral injection, local administration, intranasal administration, intrapulmonary administration, and rectal administration.

[0123] The appropriate dosage of the pharmaceutical composition of the present invention varies depending on factors such as the formulation method, administration method, patient age, weight, sex, pathological condition, food, administration time, administration route, excretion rate, and response sensitivity. A skilled physician can easily determine and prescribe a dosage effective for the desired treatment or prevention. According to a preferred embodiment of the present invention, the daily dosage of the composition of the present invention is 0.0001-100 mg / kg of the peptide derivative.

[0124] In one embodiment of the present invention, the composition of the present invention may further comprise a release controlling agent. Examples of the material used as the release controlling agent include, but are not limited to, one or more selected from butyric acid, valeric acid, caproic acid, enanthic acid, caprylic acid, pelargonic acid, capric acid, undecylic acid, lauric acid, tridecylic acid, myristic acid, pentadecylic acid, palmitic acid, heptadecylic acid, stearic acid, nonadecylic acid, arachidic acid, isocrotonic acid, oleic acid, elaidic acid, sorbic acid, linoleic acid, arachidonic acid, benzoic acid, hydroxynaphthoic acid, napadiylic acid, and pamoic acid.

[0125] The pharmaceutical composition according to the present invention may be administered in a therapeutically effective amount of a peptide derivative, for example, an effective amount for treating diabetes, specifically diabetes, beta-cell function preservation, hypertension, hyperlipidemia, obesity, non-alcoholic steatohepatitis, or degenerative neurological diseases such as Alzheimer's disease and Parkinson's disease, cardiovascular disease, tumor, brain disease, other metabolic diseases, and digestive diseases. The therapeutically effective amount of the active ingredient may be evaluated by a physician.

[0126] The pharmaceutical composition according to the present invention may be administered parenterally, for example, by subcutaneous injection. The pharmaceutical composition according to the present invention may be administered using a pre-filled syringe, for example, a pen-syringe. Such a pen-syringe may be a disposable syringe containing a single dose, or a metered dose syringe (syringe with a dose metering device) that injects only the single dose amount at a time.

[0127] In one embodiment of the present invention, the peptide is liraglutide, semaglutide, tirzepatide, dulaglutide, cotadutide, cagrilintide, cagrisema, amycretin, retatrutide, nisotirastide, pemvidutide, mazdutide, survodutide, ecnoglutide, insulin icodec, insulin degludec, somapacitan, dapiglutide, petrelintide, A pharmaceutically acceptable salt thereof is selected from the group consisting of SAR441255, NNC0165-1273, HM15136, HM15275, TE-8105, NNC0090-2746, DA-1726, Met-097i, BGM0504, VK2735 and pharmaceutically acceptable salts thereof.

[0128] In one embodiment of the present invention, the peptide delivery composition is the peptide delivery composition described above.

[0129] In one embodiment of the present invention, the disease is obesity, fatty liver disease, dyslipidemia, degenerative neurological disease, hypertension, heart failure, arteriosclerosis, diabetes, fatty liver disease, gastritis, inflammatory bowel disease, cancer, or a combination thereof.

[0130] In one embodiment of the present invention, the pharmaceutical composition is administered orally, subcutaneously, intramuscularly, or intravenously.

[0131] In one embodiment of the present invention, the pharmaceutical composition is administered repeatedly for one or more cycles.

[0132] In one embodiment of the present invention, the interval between the cycles is from 1 day to 6 months. For example, the interval between the above-mentioned speculation cycles may be 1 day to 6 months, 2 days to 6 months, 1 week to 6 months, 2 weeks to 6 months, 3 weeks to 6 months, 1 month to 6 months, 2 months to 6 months, 3 months to 6 months, 4 months to 6 months, 5 months to 6 months, 1 day to 5 months, 1 day to 4 months, 1 day to 3 months, 1 day to 2 months, 1 day to 1 month, 1 day to 2 weeks, 1 week to 5 months, 1 week to 4 months, 1 week to 3 months, 1 week to 2 months, 1 week to 1 month, 1 month to 5 months, 1 month to 4 months, or 1 month to 3 months, and is preferably 7 days to 1 month, but is not limited thereto.

[0133] Currently, peptide derivative drugs are mainly used with a dosing interval of 1 day to 1 week (e.g., liraglutide, insulin degludec, semaglutide, etc.). The pharmaceutical composition containing the peptide derivative and albumin of the present invention can extend the dosing cycle compared to when the existing peptide derivative drug is administered alone, and improve the pharmacokinetic characteristics to reduce the variation in blood concentration during repeated administration, thereby minimizing side effects (e.g., in the case of semaglutide and tirzepatide, particularly gastrointestinal side effects; nausea, vomiting, etc.), thereby improving the convenience and safety of administration for patients.

[0134] The present invention aims to improve the short half-life (administration at intervals of 1 day to 1 week) and high incidence of side effects due to rapid increase in initial blood concentration, which are the biggest limitations of existing peptide derivative drugs, and to solve the limitations of existing technology by delivering exogenous albumin together with peptide derivative drugs. Albumin is the most abundant substance transport protein in the body, and is currently utilized not only for therapeutic purposes but also as an additive in vaccines, antibody drugs, etc., so there are very few safety concerns. Recently, recombinant albumin, etc. can be utilized as a bio-derived substance with even fewer concerns about side effects.

[0135] Representative fatty acid derivative drugs of peptides such as liraglutide, semaglutide, and tirzepatide require a dose escalation period of approximately 3 months in which the dose is increased 10-fold to prevent serious side effects due to the initial rapid increase in drug concentration when administered by subcutaneous injection. According to previous studies, the dosage, blood concentration, and exposure of peptide derivative drugs are closely related to the efficacy and side effects, and in particular, the peak blood concentration (C max) is known as a key variable that can predict side effects (Diabetes Obes Metab (2018), Diabetes Obes Metab. (2018), J. Med. Chem. (2021),), Diabetes Obes Metab. 26 (2024)). Therefore, if the blood concentration below the minimum toxic concentration (MTC) can be precisely controlled through this technology, it can be seen that the optimal therapeutic effect can be achieved with minimal side effects. This ultimately means that by precisely controlling the drug in the blood within the effective concentration range, the required drug dosage can be reduced, providing greater medication convenience to patients. In addition, the dose escalation period of existing drugs can be shortened, which has the advantages of convenience and economy for patients. This can be seen as a very big advantage considering the characteristics of peptide derivative drugs that mainly treat chronic diseases.

[0136] The advantages of this technology, which can be expected by pre-loading peptide derivative drugs and albumin in vitro and then administering them as a pharmaceutical composition, can be further utilized when combined with other drug delivery technologies. For example, when an albumin-peptide derivative composition is loaded into a long-acting peptide derivative delivery technology using microspheres, it can be very helpful in maintaining a constant blood concentration by minimizing the initial burst and release amount deviation, which are drawbacks of long-acting microsphere technology. Similarly, this technology can be effectively utilized in the development of long-acting liposomes, long-acting hydrogels, microneedles, and nanoparticle drug delivery technologies because it provides an optimal release pattern.

[0137] In one embodiment of the present invention, the subject of administration of the composition or pharmaceutical composition may be a subject whose body albumin is excessively glycated. For example, it can be more useful in the case of diabetic patients or patients with albumin levels outside the normal range. In the case of diabetic patients, the albumin is glycated at a higher rate (2-30% or more) than in normal people (about 10%), and glycated albumin is unfavorable for binding to peptide derivative drugs (particularly fatty acid derivatives of peptides) administered in the body, which can result in a shortened half-life of the administered drug. Most peptide derivative drugs (particularly fatty acid derivatives of peptides) increase their half-life by binding to albumin in the body after administration. Therefore, when a peptide derivative drug and albumin are mixed and delivered in the form of a combined complex as in the present technology, the above-mentioned side effects can be reduced, efficacy reduction can be prevented, the administration cycle can be extended, and even a high therapeutic efficacy per dose can be achieved.

[0138]

[0139] The features and advantages of the present invention are summarized as follows:

[0140] (a) The present invention provides a peptide delivery composition comprising a peptide comprising a fatty acid chain and albumin.

[0141] (b) The present invention provides a method for preparing a composition for peptide delivery, comprising the step of mixing a peptide containing a fatty acid chain into a solution containing albumin.

[0142] (c) The present invention provides a pharmaceutical composition for preventing or treating metabolic diseases, neurological diseases, cardiovascular diseases, endocrine diseases, digestive diseases, oncological diseases or combinations thereof, comprising a peptide comprising a fatty acid chain and a peptide delivery composition comprising albumin.

[0143] (d) When using the composition of the present invention, compared to existing peptide drugs, the objectives of reducing peak blood concentration, delaying the time to reach peak blood concentration, reducing side effects resulting from this, maximizing efficacy by increasing the administered dose, and extending the administration cycle can be achieved.

[0144]

[0145] Figure 1 shows the ratio of unbound drug according to the mixed amount of liraglutide, semaglutide, tirzepatide, retatrutide, and cagrilintide with respect to albumin.

[0146] Figure 2 shows the pharmacokinetic profile of a single subcutaneous administration of semaglutide to rats according to the mixing amount of semaglutide with albumin.

[0147] Figure 3 shows the pharmacokinetic profile of a composition of albumin and semaglutide upon repeated subcutaneous administration in rats.

[0148] Figure 4 shows the pharmacokinetic profiles of semaglutide in rats when administered subcutaneously in combination with human serum albumin (HSA), recombinant human serum albumin (rHSA), and fatty acid free HSA.

[0149] Figure 5 shows the composition of semaglutide manufactured with polymers other than albumin and the results of pharmacokinetic profile analysis in rats in a test example.

[0150] Figure 6 shows the pharmacokinetic profile of a single subcutaneous administration in rats according to the mixed amount of Tirzepatide with respect to albumin.

[0151] Figure 7 shows the pharmacokinetic profile in rats of an albumin composition of semaglutide and Insulin Icodec when administered alone or as a single subcutaneous dose in combination.

[0152] Figure 8 shows a graph comparing the kaolin uptake and feed intake, which are indicators of nausea and vomiting, which are major digestive side effects, when an albumin and semaglutide composition is administered to rats.

[0153]

[0154] Hereinafter, the present invention will be described in more detail through examples. These examples are intended solely to illustrate the present invention more specifically, and it will be apparent to those skilled in the art that the scope of the present invention is not limited by these examples, in accordance with the gist of the present invention.

[0155]

[0156] Example

[0157] Example 1: Materials and methods.

[0158] The materials used in the present invention are as follows. The peptide derivatives as model drugs were liraglutide (Liraglutide, MediChemExpress), semaglutide (Semaglutide, Shenzhen JYMed Tech), tirzepatide (Tirzepatide, MediChemExpress), cagrilinitide (CagriChemExpress), retatrutide (Retatrutide, MediChemExpress), and insulin icodec (Insulin icodec, MediChemExpress). Albumin was human serum albumin (HSA), and Green Cross Albumin 20% injection (Green Cross, Korea) or Plasmumin. ®-25 (Grifols, Albumin USP 25% injection) was used, and recombinant human serum albumin (recombinant human serum albumin) was used. ® (Satorius) and fatty acid-free human serum albumin and rat albumin were from Merck. Monomethoxy poly(ethylene glycol)-b-poly(D,L)-lactide (mPEG-PDLLA, Samyang Holdings) and 2-hydroxypropyl beta cyclodextrin (2HP-β-CD, Merck) were used as model polymers. The centrifugal filter for pretreatment of unbound drugs was Nanosep. TM centrifugal device with Omega TM (Cytiva) was used.

[0159]

[0160] 1-1. Preparation of semaglutide-albumin composition

[0161] Human serum albumin powder was dissolved in phosphate-buffered saline (PBS) to prepare a concentration of 500 mg / mL as albumin (albumin stock solution). Semaglutide was dissolved in the albumin stock solution or diluted solution, and semaglutide and albumin were mixed to obtain the final concentrations shown in Table 2. The mixture was centrifuged at room temperature (15–25°C) for 1 hour at approximately 120 rpm to induce a binding reaction between albumin and semaglutide, and the final composition, a semaglutide-albumin complex, was obtained. The prepared test formulation was stored in a refrigerator (2–8°C) and then used in the experiment.

[0162]

[0163] 1-2. Isolation of unbound semaglutide

[0164] When a fatty acid derivative drug of a peptide electrostatically binds to albumin to form a complex, a dimer or polymer is formed, and the apparent hydrodynamic diameter becomes larger than the molecular weight of the drug itself. Therefore, when filtering using a centrifugal filter with an appropriate pore size (e.g., 10 kDa, 30 kDa, 50 kDa, 100 kDa, 300 kDa, etc.), the amount of unbound drug can be quantified by utilizing the principle that only unbound drugs with relatively low molecular weight are filtered. After testing the adsorption, permeation, and recovery rate of the drug according to the filter pore size in advance, a centrifugal filter with a permeation size of 100 kDa (Nanosep) is used. TM centrifugal device with Omega TM ) were selected. Before use, the centrifugal filter was hydrated with 1 mL of distilled water and phosphate buffer, respectively, and 400 μL of the semaglutide-albumin composition was placed in the centrifugal filter, followed by centrifugation at 20°C and 14,000 G for 10 minutes. The centrifuged solution was analyzed using HPLC by method 1-3, and the unbound drug ratio was calculated.

[0165]

[0166] 1-3. Analysis of semaglutide concentration

[0167] The composition of 1-1 or the centrifugal solution (unbound drug) of 1-2 were appropriately diluted with the mobile phase according to the concentration and analyzed using an HPLC-UV system (Agilent). The main analysis conditions are as follows.

[0168] - Column: CapcellPak UG120 (C18, 5μm, 4.6 x 150mm, Shiseido)

[0169] - Mobile phase: 45% acetonitrile aqueous solution (containing 0.1% TFA)

[0170] - Detection wavelength: UV 280 nm

[0171] - Flow rate: 1.2 mL / min

[0172] - Injection volume: 10 μL

[0173]

[0174] 1-4. Pharmacokinetic studies in rats

[0175] To study the pharmacokinetic properties of the present invention, rats, the most widely used laboratory animal and with a wealth of accumulated basic test data, were used as an animal model. Six-week-old SD rats (approximately 245–265 g) were assigned to three experimental groups after a quarantine and acclimatization period of at least one week, ensuring that the average body weight was distributed as evenly as possible. The control and test preparations were administered subcutaneously into the dorsal region of the rats using the same method, and blood samples were collected at prescribed intervals after administration. The collected blood was mixed with heparin as an anticoagulant, centrifuged at 10°C and 14,000 rpm, and the plasma was stored at -70°C or lower. The plasma was then thawed, and the drug concentration in the plasma was analyzed according to methods 1–5. No deaths were observed throughout the experiment, and no unusual symptoms related to the test substance administration were observed.

[0176]

[0177] 1-5. Analysis of blood semaglutide concentration

[0178] 40 μL of rat plasma was mixed with 200 μL of methanol (containing 50 ng / mL of Liraglutide as an internal standard), vortex-mixed for 1 minute, and centrifuged at 14,000 rpm for 5 minutes to remove proteins. 10 μL of the supernatant was injected into LC-MS / MS (SciEx).

[0179] Quantification was performed using LC-MS / MS, and the main conditions are as follows.

[0180] - Mobile phase: Gradient elution conditions of 0.3% formic acid and acetonitrile (containing 0.3% formic acid)

[0181] - Column: Capcell Pak C18 (3μm, 2 x 75 mm, Shiseido, Japan)

[0182] - Flow rate: 0.4 mL / min

[0183] - Injection volume: 10 μL

[0184] - Detection conditions: Semaglutide: 1029.4 → 136.0

[0185] Liraglutide (internal standard): 938.8 → 1064.0

[0186]

[0187] Example 2: Study on the in vitro binding properties of fatty acid derivatives of peptides and albumin.

[0188] Semaglutide, a model drug, and albumin were prepared according to the method of Example 1-1 in the ratios shown in Table 2, pretreated according to the method of Example 1-2, and analyzed according to the method of Example 1-3. The results are shown in Table 2.

[0189]

[0190] Unbound drug ratio according to semaglutide and albumin ratio Classification Semaglutide (mg / mL, nmol / mL) Albumin (mg / mL, nmol / mL) Semaglutide: Albumin ratio (weight ratio) Semaglutide: Albumin ratio (molar ratio) Unbound Semaglutide ratio (%) Comparative example 11 (243.1) 0 (0) 1:01:09 7.98 Test example 1-10.25 (60.8) 40 (601.5) 1:16 0 1:100.00 Test example 1-20.5 (121.5) 40 (601.5) 1:8 0 1:5 3.24 Test example 1-32.48 (602.9) 40 (601.5) 1:16 1:13.87 Test example 1-44.96(1205.8)40(601.5)1:81:0.51.90Test Example 2-11(243.1)0.81(12.1)1:0.81:0.0532.76Test Example 2-21(243.1)1.61(24.2)1:1.61:0.111.34Test Example 2-31(243.1)8.06(121.2)1:81:0.56.75Test Example 2-41(243.1)16.13(242.5)1:161:17.15Test Example 2-51(243.1)80.64(1214.3)1:801:50.79Test Example 2-61(243.1)161.29(2429.8)1:1611:100.00Example 2-71(243.1)250(3759.4)1:2501:15.50.00Example 2-81(243.1)500(7518.8)1:5001:310.00

[0191] As a result, the amount of unbound drug (or free drug) changed significantly as the weight ratio (molar ratio) of the model drug semaglutide and albumin changed. As the ratio of albumin increased, the ratio of unbound drug in the composition decreased significantly. When the ratio of semaglutide to albumin (molar ratio) was 1:0.05 or higher, more than 68% of the drug existed in a bound form, and approximately 32% existed in an unbound state. In addition, when the ratio of semaglutide to albumin was 1:0.5 (molar ratio) or higher, the unbound drug fell to less than 10%, and when the ratio was 1:1 (molar ratio) or higher, virtually no unbound drug was detected. Therefore, it was observed that most of the drug existed in a bound form when albumin was sufficiently present around the drug. In addition, different unbound drug ratios were observed depending on the drug concentration and albumin concentration in the composition, even though the ratio was the same.

[0192] Example 3: Single-dose pharmacokinetic (PK) study in vivo (rat)

[0193] To examine the pharmacodynamic properties of the composition in vivo, a pharmacodynamic experiment was conducted using rats as an experimental model. The control drug (semaglutide component, Wigobi) ® , Comparative Example 2) and the composition applying the present technology (Test Examples 3-1 to 3-4) were administered subcutaneously once, and the concentration of semaglutide in the blood was measured over time. The weight ratio of semaglutide and albumin in the manufactured test drug is shown in Table 3 below.

[0194]

[0195] Semaglutide and albumin concentration information by ratio of test and reference preparations Category Semaglutide (mg / mL, nmol / mL) Albumin (mg / mL, nmol / mL) Semaglutide: Albumin ratio (weight ratio) Semaglutide: Albumin ratio (molar ratio) Unbound Semaglutide ratio (%) Comparative example 20.5 (121.5) 01:01:09 7.98 Test example 3-10.5 (121.5) 40 (601.5) 1:80 1:52.04 Test example 3-20.5 (121.5) 80 (1203) 1:160 1:100.55 Test example 3-30.5 (121.5) 160 (2406) 1:320 1:200.00 Test example 3-40.5(121.5)250(3759)1:5001:30.90.00

[0196] The results of the pharmacokinetic test are shown in Figure 2 and Table 4.

[0197] C of semaglutide according to the binding ratio of albumin and semaglutide max , T max Changes in (single dose pharmacokinetics in rats). Formulation classification Dosage (mg / kg) Semaglutide: Albumin molar ratio C max * (ng / mL)Dose Normalized C max (ng / mL) ** T max (hr) Comparative Example 20.21: 01105±2745,5304.67 Test Example 3-10.31: 5994±613,31512.00 Test Example 3-20.31: 10801±932,67220.00 Test Example 3-30.31: 20524±901,74724.00 Test Example 3-40.31: 30.9439±341,46524.00

[0198] * Mean ± SD (n=3)

[0199] ** Dose Normalized C max : C max / dose (mg / kg)

[0200] As shown in Figure 2 and Table 4, the control preparation (Comparative Example 2) is Wigobi ®As a result of the experiment in which (a composition containing only semaglutide) was administered alone, T max The value was measured as 4.67 hours, C max was 1,105 ng / mL. This was similar to the results of the pharmacokinetic study of semaglutide in rats known from papers, etc. On the other hand, in the case of the test example, as the ratio of albumin bound to semaglutide increased, T max was delayed by up to 24 hours (approximately 5.4 times), and dose-normalized C max was significantly reduced to 26-60% of the control formulation. These results are consistent with the unbound drug ratio in Table 2, demonstrating the suitability of the albumin-semaglutide combination formulation as a drug delivery technology that delays initial absorption, thereby reducing side effects and extending the dosing cycle.

[0201] In summary, the test preparation was T compared to the control drug. max is delayed, C max The phenomenon of decreasing was confirmed reproducibly, and the tendency became more pronounced as the ratio of albumin increased. This shows that this technology can maintain a uniform blood concentration by minimizing fluctuations in blood concentration during repeated administration, and has the potential to increase the administration cycle.

[0202]

[0203] Example 4: Repeated-dose pharmacokinetic (PK) study in vivo (rat)

[0204] To observe the pharmacokinetic characteristics of the composition upon repeated administration in vivo, rats were used as experimental animals and a control drug (containing only semaglutide, Wigobi) was administered. ®, Comparative Example 2) or the test formulation (Test Example 3-1) using the present technology was administered subcutaneously, and the plasma concentration of semaglutide was measured over time. The administration was repeated five times at 24-hour intervals, and the pharmacokinetic analysis results of the control formulation and the test formulation after the final fifth administration are shown in Table 5 and Figure 3.

[0205] Changes in pharmacokinetic properties according to repeated administration of reference and test preparations Preparation type Dosage Pharmacokinetic parameters (n=3)T max (hr) 1 C max,ss 2 (ng / mL)C min,ss 3 (ng / mL)PTR 4 (Pear) Comparative example 20.2 mg / kg2 hr1470.43±186.80398.95±38.593.69 Test example 3-10.3 mg / kg12 hr1314.50 5 ±153.92728.98 5 ±19.601.80

[0206] 1. C max,ss : C in steady state max (mean±SD)

[0207] 2. C min,ss : C in steady state min (mean±SD)

[0208] 3. T max : Time to reach peak blood concentration (hr)

[0209] 4. PTR: Peak Trough Ratios (C max,ss / C min,ss )

[0210] 5. P < 0.01 (vs Comparative Example 2, one-way ANOVA)

[0211] As a result of analysis of pharmacokinetic characteristics at steady state after 5 repeated administrations, the test preparation (Test Example 3-1) was compared to the control drug (Wegovy ®, albumin-free, comparative example 2) T max It was confirmed that the delayed T was more than 6 times and the blood concentration deviation (PTR) was more than 50% lower. This means that even after reaching the steady state of the blood drug concentration through repeated administration, the delayed T was the same as the result of single administration. max While showing value, lower peak blood concentration (C) even at higher doses compared to the control drug max ) was shown, it was confirmed that it is a technology that can maintain a stable blood concentration through lower drug concentration variability in the steady state. This is a technology that enables precise control over the effective blood concentration range (therapeutic window) of semaglutide, and it is a technology that can maintain the same average blood concentration (C avg ) is a result showing that it can play a role in minimizing side effects by minimizing large blood concentration fluctuations (Fluctuation) and minimizing high-concentration drug exposure time.

[0212]

[0213] Example 5: Preparation and Evaluation of Various Albumin, Other Polymers, and Semaglutide Combination Compositions

[0214] In order to study the binding affinity of semaglutide to various albumins, the possibility of manufacturing compositions, and the characteristics, etc., test formulations were manufactured using the model drug semaglutide on human serum albumin, fatty acid-free human serum albumin (fatty acid-free HSA), recombinant human serum albumin (rHSA), and rat albumin, respectively, using the same method as in Example 1-1. For these compositions, the amount of unbound semaglutide drug was measured, and in vivo pharmacokinetic experiments were performed in rats.

[0215] As a result, regardless of the type of albumin, semaglutide showed a similar level of albumin binding affinity, and the unbound drug ratio in the manufactured composition was as shown in Table 6. In addition, when a single-dose pharmacokinetic test was performed on rats, similar pharmacokinetic characteristics were shown regardless of the type of albumin (Figure 4).

[0216] Meanwhile, in addition to albumin, polymers such as 2-hydroxypropyl-beta-cyclodextrin (2-HP-β-CD) and monomethoxy PEG-Poly, D, L lactic acid (mPEG-PDLLA), which are widely used as biocompatible polymers and drug delivery systems, were prepared as test formulations using the same method as in Example 1-2, and a single-dose pharmacokinetic test was conducted on rats. As a result, unlike albumin, the pharmacokinetic profile of semaglutide was not changed, and the control formulation, Wegovy ® C similar to (Comparative Example 2) max , T max (Table 6, Fig. 5).

[0217]

[0218] Evaluation of binding affinity and comparison of pharmacokinetic properties of semaglutide according to the type of excipient (albumin type, polymer, etc.) Type of formulation Type of excipient (polymer) Semaglutide: excipient ratio (molar ratio) Unbound drug (%) T max (hr) Comparative Example 3-1: 099.44.8 Test Example 4-1 Human serum albumin (Green Cross, 20%) 1: 100.524.0 Test Example 4-2 Human serum albumin (Grifols, 25%) 1: 100.124.0 Test Example 4-3 Fatty acid free HSA 1: 100.224.0 Test Example 4-4 Recombinant HSA 1: 100.220.6 Test Example 4-5 Rat Albumin 1: 101.824.0 Test Example 5-12-HP- β -Cyclodextrin 1: 1095.84.0 Test Example 5-2 mPEG-PDLLA 1: 1097.54.0

[0219]

[0220] Example 6: Preparation and evaluation of a combined composition of semaglutide and albumin at various concentrations.

[0221] The binding affinity of albumin, peptides, and peptide derivatives varies depending on the concentration of the final composition, and they exist in the form of monomers, dimers, and polymers. As a result, the amount of unbound drug changes, which in turn affects the pharmacokinetic characteristics. Therefore, the concentration of the final composition or final product immediately before administration to the patient is very important. To evaluate this effect, the amount of unbound drug was studied at various semaglutide and albumin concentrations in the composition, and the results are shown in Table 7. Human serum albumin (Green Cross, 20%) was used as the albumin.

[0222]

[0223] Unbound drug ratio according to drug and albumin concentration of the composition Classification Concentration of semaglutide (A) (mg / mL) Concentration of albumin (B) (mg / mL) A:B ratio (weight ratio) Unbound drug (%) Comparative example 4 10 1:09 5.5 Test example 6-10.10.11:147.6 Test example 6-20.10.5 1:52 5.7 Test example 6-30.11 1:10 21.4 Test example 6-40.15 1:50 17.7 Test example 6-50.11 0 1:100 10.1 Test example 6-6 10 5 0 1:51 4.2 Test example 6-7 20 5 0 1:2.5 10.8 Test example 6-8 20 10 0 1:58.7 Test example 6-9 5 0 5 0 1:10 6.8 Test example 6-101005001:55.5 Test Example 6-112005001:2.57.3

[0224] The results of a study on unbound drug in various drug and albumin concentrations showed an unbound drug ratio of less than approximately 30% in the concentration range of 0.1 to 200 mg / mL for drug and 0.5 to 500 mg / mL for albumin. Furthermore, even when the drug-to-albumin ratio was low, when the concentration of the drug itself or albumin itself was high, the unbound drug ratio was less than 15%, showing a relatively low ratio.

[0225]

[0226] Example 7: Preparation and evaluation of compositions of various peptide derivatives and albumin.

[0227] In addition to semaglutide, albumin-binding compositions were prepared and evaluated for fatty acid derivatives of various peptides, including Liraglutide, Tirzepatide, Cagrilintide, and Retatrutide.

[0228]

[0229] Example 7-1. Preparation and Evaluation of Albumin-Binding Composition of Liraglutide

[0230] Albumin was added to tertiary pH 7.4 phosphate buffered saline and stirred with a magnetic stirrer for 30 minutes to prepare albumin solutions with concentrations of 0.8, 1.6, 8, 16, 80, 160, 320, and 500 mg / mL, respectively. 10 mg of liraglutide was added to 10 mL of each concentration of albumin solution and stirred for 30 minutes to prepare albumin-bound liraglutide test formulations with different mass ratios. The prepared test formulations were stored in the refrigerator or used immediately in the experiment. For the evaluation of albumin-bound liraglutide, centrifugal filters (Nanosep® centrifugal device with Omega) were used. TM, Pall) was used to separate unbound liraglutide. Test preparations with different albumin mass ratios were placed in centrifugal filters, centrifuged at 20℃, 14,000 rcf, and 10 minutes, and the solution passing through the filter was analyzed. Unbound liraglutide was analyzed by HPLC, and the detector wavelength was 214 nm. The column used was ZORBAX Eclipse Plus-C18 (5 μm, 4.6 x 150 mm) and the column temperature was constant around 30℃. The analysis time was 50 minutes, the sample injection amount was 25 μL, the flow rate was 1.0 mL / min, the mobile phase was Mobile phase A: DW (0.1% TFA), Mobile phase B: 100% ACN (0.1% TFA), and the analysis was performed under the gradient conditions shown in Table 8 below. Unbound liraglutide according to albumin mass ratio is as shown in Table 9 below.

[0231] Liraglutide HPLC solvent conditions Time (min) Mobile phase A, % Mobile phase B, % 06535333565380100400100506535

[0232] Unbound liraglutide (%) Test results Classification Liraglutide weight (mg) Albumin weight (mg) Liraglutide: Albumin concentration ratio (mg / mL) Unbound liraglutide (%) Comparative example 5101:0105.0 Test example 7-110.81:0.825.0 Test example 7-211.61:1.613.5 Test example 7-3181:89.3 Test example 7-41161:165.1 Test example 7-51801:802.8 Test example 7-611601:1600.0 Test example 7-713201:3200.0 Test example 7-815001:5000.0

[0233]

[0234] Example 7-2. Preparation and Evaluation of Albumin-Binding Composition of Tirzepatide

[0235] Albumin was added to tertiary pH 7.4 phosphate-buffered saline and stirred with a magnetic stirrer for 30 minutes to prepare albumin solutions with concentrations of 0.8, 1.6, 8, 16, 80, 160, 320, and 500 mg / mL, respectively. 10 mg of tirzepatide was added to 10 mL of each concentration of albumin solution and stirred for 30 minutes to prepare albumin-bound tirzepatide test formulations with different mass ratios. The test formulations were stored in the refrigerator or used immediately in the experiment. For the evaluation of albumin-bound tirzepatide, centrifugal filters (Nanosep® centrifugal device with Omega TM , Pall) was used to separate unbound tirzepatide. Test preparations with different albumin mass ratios were placed in centrifugal filters, centrifuged at 20℃, 14,000 rcf, and 10 minutes, and the solution passing through the filter was analyzed. Unbound tirzepatide was analyzed by HPLC, and the detector wavelength was 220 nm. The column used was ZORBAX Eclipse Plus-C18 (5 μm, 4.6 x 150 mm) and the column temperature was constant around 30℃. The analysis time was 40 minutes, the sample injection amount was 10 μL, the flow rate was 1.0 mL / min, and the mobile phases were Mobile phase A: DW (0.1% TFA), Mobile phase B: 100% ACN (0.1% TFA), and the analysis was performed under gradient conditions as shown in Table 10 below. Unbound tirzepatide according to albumin mass ratio is as shown in Table 11 below.

[0236]

[0237] Tirzepatide gradient conditions Time (min) Mobile phase A, % Mobile phase B, % 06040304555386040406040

[0238]

[0239] Unbound Tirzepatide (%) Test result Classification Tirzepatide weight (mg) Albumin weight (mg) Tirzepatide: Albumin concentration ratio (mg / mL) Unbound Tirzepatide (%) Comparative example 6101:0111.0 Test example 8-110.81:0.827.0 Test example 8-211.61:1.618.2 Test example 8-3181:813.3 Test example 8-41161:167.7 Test example 8-51801:802.0 Test example 8-611601:1600.0 Test example 8-713201:3200.0 Test example 8-815001:5000.0

[0240] Example 7-3. Preparation and Evaluation of Albumin-Binding Composition of Cagrilintide

[0241] Albumin was added to tertiary pH 7.4 phosphate buffered saline and stirred with a magnetic stirrer for 30 minutes to prepare albumin solutions with concentrations of 0.8, 1.6, 8, 16, 80, 160, 320, and 500 mg / mL, respectively. 10 mg of kagrillintide was added to 10 mL of each concentration of albumin solution and stirred for 30 minutes to prepare albumin-bound kagrillintide test formulations with different mass ratios. Each test formulation was stored in the refrigerator or used immediately in the experiment. For the evaluation of albumin-bound kagrillintide, a centrifugal filter (Nanosep® centrifugal device with Omega TM, Pall) was used to separate unbound kagrillintide. Test preparations with different albumin mass ratios were placed in centrifugal filters, centrifuged at 20℃, 14,000 rcf, and 10 minutes, and the liquid passing through the filter was analyzed. Unbound kagrillintide was analyzed by HPLC, and the detector wavelength was 214 nm. The column used was ZORBAX Eclipse Plus-C18, 5 um, 4.6 x 150 mm, and the column temperature was constant around 30℃. The analysis time was 50 minutes, the sample injection amount was 25 μL, the flow rate was 1.0 mL / min, and the mobile phase was Mobile phase A: DW (0.1% TFA), Mobile phase B: 100% ACN (0.1% TFA). The analysis was performed under the gradient conditions shown in Table 12 below. The results of unbound kagrillintide according to the albumin mass ratio are shown in Table 13.

[0242]

[0243] Kagrilintide gradient conditions Time (min) Mobile phase A, % Mobile phase B, % 06535333565380100400100506535

[0244]

[0245] Unbound Kagrilintide (%) Test result Classification Kagrilintide weight (mg) Albumin weight (mg) Kagrilintide: Albumin concentration ratio (mg / mL) Unbound Kagrilintide (%) Comparative example 7 10 1:099.5 Test example 9-1 10.8 1:0.83 0.5 Test example 9-2 11.6 1:1.62 1.8 Test example 9-3 18 1:812.0 Test example 9-4 116 1:166.7 Test example 9-5 18 0 1:801.8 Test example 9-6 116 0 1:1600.0 Test example 9-7 132 0 1:3200.0 Test example 9-8 150 0 1:5000.0

[0246] Example 7-4. Preparation and Evaluation of Albumin-Binding Composition of Retatrutide

[0247] Albumin was added to pH 7.4 phosphate-buffered saline and stirred with a magnetic stirrer for 30 minutes to prepare albumin solutions with concentrations of 0.8, 1.6, 8, 16, 80, 160, 320, and 500 mg / mL, respectively. 10 mg of letatrutide was added to 10 mL of each concentration of albumin solution and stirred for 30 minutes to prepare albumin-bound letatrutide test formulations with different mass ratios. The test formulations were stored in the refrigerator or used immediately in the experiment. For the evaluation of albumin-bound letatrutide, a centrifugal filter (Nanosep® centrifugal device with Omega TM , Pall) was used to separate unbound letatrutide. Test preparations with different albumin mass ratios were placed in centrifugal filters, centrifuged at 20℃, 14,000 rcf, and 10 minutes, and the solution passing through the filter was analyzed. Unbound letatrutide was analyzed by HPLC, and the detector wavelength was 214 nm. The column used was ZORBAX Eclipse Plus-C18 (5 μm, 4.6 x 150 mm) and the column temperature was constant around 30℃. The analysis time was 50 minutes, the sample injection amount was 25 μL, the flow rate was 1.0 mL / min, and the mobile phase was Mobile phase A: DW (0.1% TFA), Mobile phase B: 100% ACN (0.1% TFA). The gradient conditions shown in Table 14 below were used for analysis. The results of unbound retatrutide according to the albumin mass ratio are shown in Table 15 below.

[0248]

[0249] Letatrutide gradient conditions Time (min) Mobile phase A, % Mobile phase B, % 06535333565380100400100506535

[0250]

[0251] Unbound Retatrutide (%) Test Results Classification Retatrutide Weight (mg) Albumin Weight (mg) Retatrutide: Albumin Concentration Ratio (mg / mL) Unbound Retatrutide (%) Comparative Example 8 10 1:096.3 Test Example 10-1 10.8 1:0.83 2.6 Test Example 10-2 11.6 1:1.6 19.8 Test Example 10-3 18 1:813.8 Test Example 10-4 116 1:167.2 Test Example 10-5 18 0 1:803.8 Test Example 10-6 116 0 1:1600.0 Test Example 10-7 132 0 1:3200.0 Test Example 10-8 150 0 1:5000.0

[0252] In conclusion, K calculated from the unbound rate according to the albumin ratio of the model drugs Semaglutide, Tirzepatide, Liraglutide, Cargrilintide, and Retatrutide 50 , K 30 (The ratios with unbound rates of 50% and 30%, respectively) are calculated as shown in Table 16. The ratio (weight ratio) of 50% or more binding by binding of fatty acid derivatives of peptides to albumin was in the range of 1:0.41 to 1:0.58, and the weight ratio of 70% or more binding (unbound rate of 30% or less) was observed in the range of 1:0.79 to 1:1.11.

[0253] Calculation results of the composition ratio of the albumin composition of the model drug with an unbound rate of 50 and 30%. Model drug Name drug: Albumin Molar ratio and concentration (weight) ratio (mg / mL) Unbound rate 50% Unbound rate 30% Semaglutide 1: 0.471: 0.86 Tirzepatide 1: 0.561: 1.01 Liraglutide 1: 0.471: 0.87 Cargrilintide 1: 0.581: 1.11 Retatrutide 1: 0.561: 1.08

[0254] Example 8: Pharmacokinetic study of Tirzepatide in vivo (rat)

[0255] In addition to semaglutide, Tirzepatide, a representative peptide fatty acid derivative drug, was prepared by applying Example 1-1, and a pharmacokinetic test was performed using rats as experimental animals in a similar manner to Example 3. The dosage was 0.2 mg / kg as Tirzepatide, and a single subcutaneous administration was performed. As a result, similar to semaglutide, T max Increase of C max A decrease was observed, and details are shown in Table 17 and Fig. 6.

[0256]

[0257] Results of the rat pharmacokinetic study of tirzepatide-albumin composition (n=3) Classification Tirzeptaide: Albumin Weight Ratio Tmax (hr) Cmax * (ng / mL) Comparative Example 91: 012.0585.1 Test Example 11-11: 10024.0 (twice the comparative example) 486.4 (83.1% of the comparative example) Test Example 11-21: 20024.0 (twice the comparative example) 479.5 (81.9% of the comparative example)

[0258] *Mean±SD

[0259]

[0260] Example 9: Pharmacokinetic study of combination therapy of Semaglutide and Insulin Icodec in vivo (rat)

[0261] In order to evaluate the pharmacodynamic effects when manufacturing a composition of two or more fatty acid derivatives and albumin, a combination drug was manufactured using semaglutide and insulin Icodec as model drugs. The manufacturing method was the same as Example 1-1, and the composition was manufactured so that semaglutide was 1 mg / mL, insulin Icodec was 10 mg / mL, and albumin was 200 mg / mL. As a comparative example, a control drug, Wigobi, was used. ®As a test example, a complex formulation containing albumin and a complex formulation not containing albumin were prepared, and the concentration of unbound semaglutide was analyzed for each (Table 18). The prepared control formulation (Wigobi) ® ) or the test preparation was administered subcutaneously as semaglutide at a single dose of 0.2 mg / kg to three rats, and blood samples were collected at appropriate intervals for 72 hours, and the drug concentration in the plasma was analyzed using LC-MS / MS in the same manner as in Example 1-5.

[0262] As a result of analyzing the ratio of unbound drugs and pharmacokinetic characteristics according to the manufacturing of combination drugs, in the case of the combination drug of semaglutide and insulin icodec, the concentration of unbound semaglutide increased rapidly as the amount of albumin decreased, and in particular, in the pharmacokinetic test, in the case of the combination drug without albumin binding, T was higher than that of semaglutide alone. max is short, C max In other words, in the case of fatty acid derivatives of peptides, when preparing a complex formulation, the drug with a relatively high unbound ratio was rapidly absorbed through the capillaries upon subcutaneous injection, resulting in a rapid increase in blood concentration. This result may lead to an increase in side effects, which may limit the use of the drug.

[0263]

[0264] Semaglutide, insulin icodec combination preparation, unbound ratio, pharmacokinetic test results (n=3) Classification Semaglutide: insulin icodec: albumin weight ratio Unbound semaglutide (%) T max (hr)C max * (ng / mL) Comparative Example 101: 0: 098.84.0954.4 Test Example 12-11: 10: 093.32.0996.0 Test Example 12-21: 10: 2048.5--Test Example 12-31: 10: 2000.624.0604.7

[0265] * Mean±SD In conclusion, the present invention, which is a fatty acid derivative composition of a peptide combined with albumin, is judged to be able to maximize the therapeutic effect by preventing a rapid increase in the blood concentration of the drug, especially when administered as a combination drug, thereby reducing side effects, improving medication compliance, and increasing the continuity of drug administration.

[0266]

[0267] Example 10: In vivo adverse effect test of semaglutide-albumin composition

[0268] The fatty acid derivative composition of the albumin-bound peptide according to the present invention has the advantage of reducing side effects, particularly by preventing a rapid increase in blood concentration after administration. To evaluate this effect, the pica behavior test was performed in a rodent model. Rodents do not experience nausea or emesis, and when such gastrointestinal discomfort is experienced, they tend to ingest kaolin instead of food. This has been well-established as a test model (Methods Find Exp Clin Pharmacol 2002, Neuropharmacol 2012).

[0269] Male SD rats, approximately 245-255 g and 6 weeks old, were used as animal models. In addition to general acclimatization, feed and kaolin were placed in separate compartments within the cage for 5 days prior to the experiment to acclimatize the rats to the feed and kaolin. The finally selected rats were divided into groups of 7 rats and fasted for 18 hours before the experiment. On the test day, the placebo (vehicle), control agent (Wigobi®, 0.2 mg / kg), and test agent (low dose (0.2 mg / kg), high dose (0.4 mg / kg)) were administered subcutaneously once, and the food intake and kaolin intake were evaluated after 4, 8, 12, 24, 36, 48, and 72 hours, respectively (Table 19, Fig. 8).

[0270]

[0271] Kaolin intake test according to administration of semaglutide-albumin composition (n=7) Classification (dosage) Semaglutide: Albumin weight ratio Cumulative feed intake (g) Cumulative kaolin intake (g) 8hr24hr8hr24hr Comparative example 11-1 (vehicle) 0: 20013.9±0.931.2±4.60.12±0.020.15±0.05 Comparative example 11-2 (0.2mg / kg) 1: 08.5±0.310.4±0.51.45±0.162.42±0.38 Test example 13-1 (0.2mg / kg) 1: 2009.7±0.614.3±1.10.72±0.12 * 1.65±0.28 * Test Example 13-2 (0.4 mg / kg) 1: 2008.6 ± 0.4 9.8 ± 1.3 0.89 ± 0.16 ** 2.44±0.42

[0272] * P<0.001 (vs Comparative Example 11-2, one-way ANOVA)

[0273] ** P<0.01 (vs Comparative Example 11-2, one-way ANOVA)

[0274] As a result of the experiment, in the vehicle group (administered with albumin solution), there was no significant change in feed intake or kaolin intake, and normal activity was observed. In other words, kaolin intake was almost nonexistent, and feed intake was approximately 31 g / day, which is the normal intake rate. However, in the control group (Wigobi) ® ) showed a significant decrease in feeding activity immediately after administration, and the kaolin intake increased rapidly proportionally. In particular, the kaolin intake for 8 hours and 24 hours after administration showed a statistically significant decrease in the test example. After about 48 hours, the feeding amount and speed almost returned to normal, and the kaolin intake also decreased. This was evaluated as a good demonstration of the effect of the invention technology that slows down the initial drug absorption. The high-dose test group (0.4 mg / kg, Test Example 13-2) showed a relatively decrease in feeding amount and an increase in kaolin intake compared to the low-dose test group. However, despite the high dose, the control group (Wegobi) ®, compared to Comparative Example 11-2), the amount of kaolin intake decreased over the initial 8 hours, and it was evaluated that side effects were significantly reduced due to the delay in the initial absorption rate despite the high dose. In conclusion, it is judged that the development technology of the present invention can prevent a rapid increase in the initial blood concentration by slowing down the initial absorption rate of a peptide derivative drug, and alleviate the resulting side effects.

[0275]

[0276] In summary, the composition of the present invention including albumin has a lower C at the same dosage compared to a fatty acid derivative of a peptide (control formulation) that does not include albumin. max and delayed T max By achieving this, we were able to significantly reduce side effects due to the initial rapid drug release, and confirmed that there is a possibility that the administration cycle can be extended beyond the existing once-a-week period. In particular, the peak trough ratio of the drug in the blood at steady state during repeated administration could be reduced to less than 50% compared to existing drugs, and through this, there is a very high possibility of achieving maximization of therapeutic effect through low incidence of side effects, improved patient compliance, and reduced possibility of discontinuation of administration. In particular, since side effects are low, it means that the dosage can be increased more than existing drugs, which means that the efficacy can be maximized.

[0277] In addition, the pharmacokinetic improvement characteristics of albumin were relatively verified through comparison with substances widely used to improve drug delivery efficacy. A mixed formulation of semaglutide and mPEG-PDLLA, which has been verified to form polymeric micelles (approximately 20 nm in size), and Hydroxypropyl β-Cyclodextrin (2-HP-β-CD), which is widely used as an additive, were prepared, and the pharmacokinetic characteristics in rats were compared and verified with the test drug (albumin-semaglutide mixed formulation). As can be seen in Figure 5, there was no difference in the pharmacokinetic characteristics of polymeric micelles (mPEG-PDLLA) and 2-HP-β-CD from the semaglutide control drug, and in particular, T max No delay was observed. On the other hand, in the case of the test drug of the present invention, a significant T max Delayed release (delayed release effect) and a gradual increase in blood concentration (C max Clear effects such as reduction of ) were observed. In conclusion, it was confirmed that the pharmacokinetic profile of fatty acid derivatives of peptides can be significantly improved when utilizing the composition under optimal conditions based on the biological efficacy of albumin.

[0278]

[0279] The present invention was developed with the goal of maximizing efficacy and minimizing side effects by utilizing exogenous albumin, which possesses excellent biocompatibility, as a delivery vehicle. This technology, as an efficient drug delivery technology, maximizes efficacy and safety by preventing the initial rapid increase in blood concentration of peptide fatty acid derivative drugs, reducing the resulting side effects, and extending the dosing cycle.

Claims

1. Composition for peptide delivery comprising a peptide containing a fatty acid chain and albumin: (a) the concentration of the peptide is 0.1 mg / mL to 500 mg / mL; and (b) The concentration of the albumin is 0.1 mg / mL to 500 mg / L.

2. A composition for peptide delivery, wherein the weight ratio of the peptide and albumin in the first paragraph is 1:0.6 to 1:

500.

3. A composition for peptide delivery, wherein the molar ratio of the peptide and albumin in paragraph 1 is 1:0.05 to 1:

30.

4. A composition for peptide delivery in the first paragraph, wherein the unbound peptide among the peptides is 30 wt% or less of the total peptide weight.

5. A composition for delivering a peptide, wherein the albumin in the first paragraph is composed of albumin bound to a peptide, free albumin, or a combination thereof.

6. A composition for peptide delivery according to claim 1, wherein the peptide is composed of 15 to 250 amino acids.

7. A composition for peptide delivery according to claim 1, wherein the fatty acid chains are 1 to 5.

8. A composition for peptide delivery, wherein the number of carbon atoms in the fatty acid chain included in the peptide in the first paragraph is 8 to 30.

9. In the first paragraph, the peptide is liraglutide, semaglutide, tirzepatide, dulaglutide, cotadutide, cagrilintide, cagrisema, amycretin, retatrutide, nisotirastide, pemvidutide, mazdutide, survodutide, ecnoglutide, insulin icodec, insulin degludec, somapacitan, dapiglutide, petrelintide, A composition for peptide delivery, wherein the composition is selected from the group consisting of SAR441255, NNC0165-1273, HM15136, HM15275, TE-8105, NNC0090-2746, DA-1726, Met-097i, BGM0504, VK2735 and pharmaceutically acceptable salts thereof.

10. A composition for peptide delivery according to claim 1, wherein the albumin is selected from the group consisting of human serum albumin (HSA), recombinant human serum albumin (rHSA), bovine serum albumin (BSA), ovalbumin (OVA), and combinations thereof.

11. A method for producing a composition for peptide delivery, comprising the step of mixing a peptide containing a fatty acid chain into a solution containing albumin.

12. A method for producing a composition for peptide delivery, wherein the weight ratio of the peptide and albumin in the composition is 1:0.8 to 1:500 in the 11th paragraph.

13. A method for producing a composition for peptide delivery, wherein the concentration of the peptide and albumin in the composition in claim 11 is 0.1 mg / mL to 500 mg / mL.

14. A pharmaceutical composition for preventing or treating metabolic diseases, neurological diseases, cardiovascular diseases, endocrine diseases, digestive diseases, oncological diseases or combinations thereof, comprising a peptide comprising a fatty acid chain and a peptide delivery composition comprising albumin: The above peptide is a glucagon-like peptide-1 receptor agonist (GLP-1 RA), insulin icodec, or a combination thereof.

15. In the 14th paragraph, the peptide is liraglutide, semaglutide, tirzepatide, dulaglutide, cotadutide, cagrilintide, cagrisema, amycretin, retatrutide, nisotirastide, pemvidutide, mazdutide, survodutide, ecnoglutide, insulin icodec, insulin degludec, somapacitan, dapiglutide, petrelintide, A pharmaceutical composition selected from the group consisting of SAR441255, NNC0165-1273, HM15136, HM15275, TE-8105, NNC0090-2746, DA-1726, Met-097i, BGM0504, VK2735 and pharmaceutically acceptable salts thereof.

16. A pharmaceutical composition according to claim 14, wherein the peptide delivery composition is a composition according to any one of claims 1 to 10.

17. A pharmaceutical composition according to claim 14, wherein the disease is obesity, fatty liver disease, dyslipidemia, degenerative neurological disease, hypertension, heart failure, arteriosclerosis, diabetes, fatty liver disease, gastritis, inflammatory bowel disease, cancer, or a combination thereof.

18. A pharmaceutical composition according to claim 14, wherein the pharmaceutical composition is administered orally, subcutaneously, intramuscularly, or intravenously.

19. A pharmaceutical composition according to claim 14, wherein the pharmaceutical composition is administered repeatedly for one cycle or more.

20. A pharmaceutical composition according to claim 19, wherein the interval between the cycles is from 1 day to 6 months.

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