Pharmaceutical formulation of glucagon-like peptide-1 receptor agonist peptides suitable for sublingual administration

Abstract

The present disclosure relates to a pharmaceutical formulation suitable for sublingual administration for use in the treatment of obesity, type II diabetes, or related diseases. The pharmaceutical formula comprises 5 to 10 milligrams of glucagon-like peptide-1 receptor agonist peptide (GLP-1 RA-P) which is dissolved in 1.5 to 2.0 milliliters of a buffer solution, and the pH (Potential of Hydrogen) value of the pharmaceutical formula is 5.5 to 7.5. The disclosure also discloses a method for treating obesity, type II diabetes, or related diseases. The method comprises administering the pharmaceutical formulation of the present disclosure to a subject in a sublingual administration, and allowing the pharmaceutical formulation to be continuously located sublingually of the subject for 5-10 minutes, where the pharmaceutical formulation has a bioavailability for GLP-1 RA-P of 2-10% compared to the administration to the subject through subcutaneous injection.

Description

[0001] Sequence list information reference

[0002] This application also includes, together with, an electronic sequence list file named 'P4375_SEQ_AF', created on January 10, 2025, with a file size of 4kB. The information contained in this electronic sequence list file is incorporated herein by reference.

[0003] Cross-reference to related applications

[0004] This application claims the full benefit of U.S. Provisional Application No. 63 / 626,512, filed on January 29, 2024, the contents of which are incorporated herein by reference. Background of the Invention 1. Technical Field

[0007] This disclosure pertains to the field of pharmaceutical formulations. Specifically, this disclosure relates to a pharmaceutical formulation suitable for sublingual administration and its use in the treatment of conditions related to overweight or hyperglycemia.

[0008] 2. Previous Technical Description

[0009] Diabetes and obesity are major diseases facing humanity.

[0010] Diabetes mellitus is an endocrine disorder characterized by high blood sugar, glucose in urine, and elevated levels of glycated hemoglobin (HbA1c) in the blood. It can lead to a wide range of macrovascular complications, including coronary artery disease, stroke, and peripheral artery disease, as well as small vessel complications affecting the eyes, kidneys, and nerves. Common symptoms of diabetes include high blood pressure, heart disease, fatty liver, and non-alcoholic steatohepatitis (NASH), as well as chronic ulcers in the legs and feet. Approximately 10-15% of diabetes cases are type 1 diabetes, caused by the pancreas's inability to produce enough insulin, and can be treated with insulin. In contrast, about 80% of all diabetes cases are type 2 diabetes, caused by insulin resistance. Treatment for type 2 diabetes generally involves metformin, GLP-1 receptor agonists, and lifestyle modifications such as exercise and weight management.

[0011] In recent years, the medical community and government healthcare institutions have gradually recognized that obesity is not only a lifestyle issue but also a major disease. Obesity is defined as a body mass index (BMI) greater than 30 kg / m², and is associated with a higher risk of type 2 diabetes, cardiovascular disease, obstructive sleep apnea, osteoarthritis, depression, and certain types of cancer. The prevalence of obesity and diabetes (usually not as comorbidities) is rising rapidly, affecting 10-30% of the total population in many countries. This situation creates significant demands on healthcare resources and a heavy economic burden.

[0012] GLP-1 inhibitors (RAs) are primarily used to treat type 2 diabetes and obesity.

[0013] Glucagon-like peptide-1 (GLP-1) is produced and secreted by L cells in the enteroendocrine system of the small intestine and certain neurons in the brainstem after food consumption. The initially produced GLP-1 (1-37) undergoes protein hydrolysis to produce the biologically active GLP-1 (7-36) amide and GLP-1 (7-37). GLP-1 plays important roles in regulating various organs, including the pancreas, stomach, liver, muscles, adipose tissue, kidneys, brain, and bones. One of GLP-1's main functions is to work with the gastric inhibitory peptide GIP (also known as glucose-dependent peptide) to stimulate insulin secretion and inhibit the release of glucagon from the pancreas, thereby lowering blood glucose levels. Furthermore, GLP-1 inhibits gastric emptying, gastric acid secretion, and gastric motility, collectively reducing appetite.

[0014] Many peptides that act as GLP-1 receptor agonists (referred to herein as GLP-1 RA-Ps) have been developed and approved for the treatment of type 2 diabetes and obesity. However, apart from semaglutide (brand name Rybelsus), which is an oral formulation, these peptides are all administered via subcutaneous injection. Many patients in the large population affected by type 2 diabetes or obesity are afraid of injections. An injection-free administration method could improve patient acceptance and compliance, especially for new patients. Such a delivery system would facilitate the promotion of self-administration or home use. Among all injection-free administration methods for therapeutic peptides and proteins (including GLP-1 RA-Ps), semaglutide is representative of this technology because it allows for the administration of an effective amount of Rybelsus orally once daily. However, Rybelsus has low bioavailability, only about 0.1-1%, and its absorption needs to be improved under optimized conditions.

[0015] Bioavailability is key to the feasibility of administering peptide drugs via sublingual administration.

[0016] For peptide and protein drugs, intravenous, subcutaneous, and intramuscular injections are the standard methods for parenteral administration. Because peptides and proteins are digested by various digestive enzymes in the stomach, oral administration is ineffective. Only a few peptide drugs are developed for oral or sublingual administration by crossing the mucosal epithelium.

[0017] Generally, the bioavailability of a drug administered to an individual can be defined as the percentage (%) of the administered drug remaining in the body of the individual after a given observation period. When drugs are administered via intradermal, subcutaneous, or intramuscular injection, relatively high bioavailability is usually achieved, sometimes even approaching 100%. This depends on the extent to which the drug is digested or inactivated in tissues as it diffuses from the injection site into the bloodstream. The relatively high bioavailability associated with parenteral administration also applies to protein and peptide drugs.

[0018] Many patients fear injections, and injections negatively impact patient compliance, thus desiring oral or sublingual administration. The convenience and comfort of oral and sublingual administration enhance medication adherence. However, only a very limited number of protein and peptide drugs are approved for this route of administration. One notable exception is oxytocin and synthetic desmopressin, which can be administered sublingually in addition to injection. Smegglutide has been approved as an oral tablet, designed with a unique formulation and specific timing of food and water intake. Furthermore, an oral formulation of human growth hormone is currently undergoing phase II clinical trials in humans. These proteins or peptides are hormones that function at low concentrations, contributing to their functional bioavailability.

[0019] Sublingual formulations of oxytocin showed a 10-fold difference in bioavailability in treated human individuals, ranging from 0.007% to 0.07% (see De Groot AN et al, J Pharm Pharmacol (1995) 47: 571-5). The absolute bioavailability of desmopressin administered sublingually varied between 0.21% and 0.31% in treated human individuals (see Electronic Medicines Compendium. Desmopressin. 120 microgram Sublingual Tablets. Molecule Pharma Ltd., Aug. 30, 2023; Oiso Y, et al. J Clinical Endocrinology & Metabolism (2013) 98 (10), 3958-3967). In human individuals receiving treatment, the bioavailability of the oral form of smegglutide (trade name: Rybelsus) is between 0.4% and 1%, and requires suitable conditions for absorption (see Novo Nordisk Inc. Rybelsus Prescribing Information, October 2019).

[0020] The need to improve sublingual delivery of GLP-1 RA-P

[0021] Currently, only two peptide hormones (oxytocin and desmopressin) have been developed, marketed, and used clinically via sublingual delivery. In contrast, sublingual delivery methods for other peptide and protein drugs have not yet been developed. Therefore, there is an urgent need in the art for an improved oral formulation of GLP-1 RA-P for the treatment of obesity or type II diabetes, wherein the sublingual formulation of GLP-1 RA-P disclosed herein can achieve a bioavailability of 7-10%. Summary of the Invention

[0022] This disclosure is based on the discovery that GLP-1 RA-P can be successfully delivered via the sublingual mucosa. Accordingly, this disclosure provides a pharmaceutical formulation for the treatment of obesity, type II diabetes, or related diseases via sublingual administration.

[0023] According to one embodiment of this disclosure, the pharmaceutical formulation comprises: 5-10 mg of glucagon-like peptide-1 receptor agonist peptide (GLP-1 RA-P) was dissolved in 1.5-2.0 mL of buffer solution; in GLP-1 RA-P is selected from the group consisting of TE-8105, semaglutide and tirzepatide, among which TE-8105 has the structure shown in Figure 1; The pH value of the pharmaceutical formulation is between 5.5 and 7.5, and it is suitable for sublingual administration; and Compared to administration to individuals via subcutaneous injection, the pharmaceutical formulation achieves a bioavailability of 2-10% for GLP-1 RA-P.

[0024] According to one embodiment of this disclosure, the pharmaceutical formulation is in the form of a liquid, tablet, or gel.

[0025] According to certain embodiments of this disclosure, the pharmaceutical formulation is in liquid form and contains 5-10 mg of TE-8105 dissolved in 1.5-2.0 mL of buffer solution.

[0026] According to other embodiments of this disclosure, the pharmaceutical formulation is in liquid form and contains 5-6 mg of smegglutinin dissolved in 1.5-2.0 mL of buffer solution.

[0027] According to another embodiment of this disclosure, the pharmaceutical formulation is in liquid form and contains 5-10 mg of telpoeptide dissolved in 1.5-2.0 mL of buffer solution.

[0028] According to an alternative embodiment of this disclosure, the pharmaceutical formulation further comprises 0.1% (w / v) peppermint oil.

[0029] According to the optional embodiments disclosed herein, the pharmaceutical formulation further comprises 5-10% gelatin, and the pharmaceutical formulation is in the form of a soft gel.

[0030] According to an alternative embodiment of this disclosure, the pharmaceutical formulation further comprises 1-10% crospovidone, and the pharmaceutical formulation is in the form of tablets.

[0031] According to the embodiments disclosed herein, the diseases associated with obesity and type II diabetes are overweight, fatty liver, non-alcoholic fatty liver disease, diabetic cardiomyopathy, or coronary atherosclerotic heart disease.

[0032] In another embodiment, this disclosure aims to provide a method for treating obesity, type II diabetes, or related diseases. The method comprises: An effective amount of a pharmaceutical formulation is administered to the individual sublingually and the formulation is maintained under the individual's tongue for 5-10 minutes. The pharmaceutical formulation contains glucagon-like peptide-1 receptor agonist peptide (GLP-1 RA-P). in, The GLP-1 RA-P is selected from the group consisting of TE-8105, smegglutinin and telpokinin, wherein TE-8105 has the structure shown in Figure 1. The pH value of the pharmaceutical formulation is between 5.5 and 7.5; The pharmaceutical formulation is administered to the individual at a frequency of once daily, twice daily, once every two days, once weekly, or twice weekly; and Compared to administration to individuals via subcutaneous injection, the pharmaceutical formulation achieves a bioavailability of 2-10% for GLP-1 RA-P.

[0033] According to the embodiments disclosed herein, the pharmaceutical formulation is in the form of a liquid, tablet, or gel.

[0034] According to certain embodiments of this disclosure, the pharmaceutical formulation is in liquid form and contains 5-10 mg of TE-8105 dissolved in 1.5-2.0 mL of buffer solution, and is administered to the individual at a frequency of twice daily, once daily, once every two days, once weekly, or twice weekly.

[0035] According to certain embodiments of this disclosure, the pharmaceutical formulation is in liquid form and contains 5-6 mg of smegglutinin dissolved in 1.5-2.0 mL of buffer solution, and is administered to the individual at a frequency of twice daily, once daily, or once every two days.

[0036] According to certain embodiments of this disclosure, the pharmaceutical formulation is in liquid form and contains 5-10 mg of telpolide dissolved in 1.5-2.0 mL of buffer solution, and is administered to the individual twice daily.

[0037] According to an alternative embodiment of this disclosure, the pharmaceutical formulation further comprises 0.1-0.5% (w / v) peppermint oil.

[0038] According to an optional embodiment of this disclosure, the pharmaceutical formulation further comprises 5-10% (w / v) gelatin, in the form of a soft gel.

[0039] According to the optional embodiments disclosed herein, the pharmaceutical formulation further comprises 1-10% crospovidone and is in the form of tablets.

[0040] According to the embodiments disclosed herein, the diseases associated with obesity and type II diabetes are overweight, fatty liver, non-alcoholic fatty liver disease, diabetic cardiomyopathy, or coronary atherosclerotic heart disease.

[0041] In all embodiments of this disclosure, the individual is a human being.

[0042] After reading the following embodiments, those skilled in the art will easily understand the basic spirit and other inventive objectives of the present invention, as well as the technical means and implementation methods adopted by the present invention.

[0043] Simple Explanation of the Diagram

[0044] To make the above and other objects, features, advantages and embodiments of the present invention more apparent and understandable, the accompanying drawings are described below: Figure 1 is a schematic diagram of the structure of TE-8105, which is disclosed herein; Figure 2 compares serum glucose levels in db / db mice treated with once-daily subcutaneous injection (SC) and twice-daily sublingual administration (SL) for 3 days. (A) Changes in serum glucose levels over time; (B) Serum glucose levels at three time points before treatment with different SL doses, compared with those treated with a single SC dose; (C) Area under the curve (AUC) of serum glucose levels over 168 hours; analysis was performed using two-dimensional ANOVA. P < 0.05; P < 0.01; P < 0.0010; P < 0.00001; Figure 3 is a three-line graph showing the change of blood glucose levels in the three mice of each group over time, based on Figure 2. Figure 4 illustrates the additional and cumulative efficacy of repeated sublingual administration of different doses of TE-8105 to db / db mice. (A) Comparison of the efficacy of six doses of TE-8105 administered sublingually once daily in hypoglycemic conditions at doses of 100 nmol / kg, 150 nmol / kg, and 250 nmol / kg, with the efficacy of a single subcutaneous injection of TE-8105 at doses of 30 nmol / kg; (B) Serum glucose levels after two single SL administrations, with a third glucose level determination performed before the third administration; (C) AUC of serum glucose levels over a 168-hour timeframe. Figure 5 compares the blood glucose levels in db / db mice that received semaglutide twice subcutaneously and twice daily for three days. (A) Changes in blood glucose levels over time; (B) Comparison of blood glucose levels at the first three time points after SL administration of different doses with those after a single SC administration; (C) AUC of blood glucose over a 144-hour time span. Figure 6 compares the serum glucose levels in db / db mice after two subcutaneous injections and twice-daily administration of telpolide for three days. (A) Changes in serum glucose levels over time; (B) Comparison of serum glucose levels at the first three time points after administration of different doses with SL and after one SC administration; (C) AUC of serum glucose over a 144-hour time span. Figure 7 illustrates the serum glucose levels in mice administered TE-8105 via sublingual or oral administration at a rate of db / db. (A) Changes in serum glucose levels over time; (B) AUC of serum glucose over a 192-hour timeframe; Figure 8 illustrates the body weight of db / db mice administered TE-8105 sublingually or orally. (A) Change in body weight of db / db mice over time; (B) AUC of body weight over a 192-hour timeframe; Figure 9 illustrates the food intake of db / db mice administered TE-8105 via sublingual or oral administration. (A) Changes in food intake over time; (B) AUC of food intake over a 192-hour timeframe; Figure 10 illustrates the water intake of db / db mice administered TE-8105 via sublingual or oral administration. (A) Changes in water intake of db / db mice over time; (B) AUC of water intake over a 168-hour timeframe; Figure 11 illustrates the serum glucose levels in db / db mice after sublingual administration of different concentrations of TE-8105 for 5 days. (A) Changes in serum glucose levels over time and the AUC of serum glucose levels over a 216-hour timeframe; and (B) Changes in serum glucose levels over 72 hours and the AUC of serum glucose levels over a 72-hour timeframe; Figure 12 illustrates the body weight of db / db mice after 5 days of sublingual administration of different concentrations of TE-8105. (A) Changes in body weight over time and AUC of body weight over a 216-hour timeframe; and (B) Changes in body weight over 72 hours and AUC of body weight over a 72-hour timeframe; Figure 13 illustrates the food intake of db / db mice administered different concentrations of TE-8105 sublingually for 5 days. (A) Changes in food intake over time and the AUC of food intake over a 216-hour timeframe; and (B) Changes in food intake over 72 hours and the AUC of food intake over a 72-hour timeframe. Figure 14 illustrates the water intake of db / db mice after 5 days of sublingual administration of different concentrations of TE-8105. (A) Changes in water intake over time and the AUC of water intake over a 216-hour period; and (B) Changes in water intake over 72 hours and the AUC of water intake over a 72-hour period. Figure 15 illustrates the serum glucose levels in db / db mice after 5 days of once-daily sublingual administration of TE-8105. (A) Changes in serum glucose levels over time and the AUC of serum glucose levels over a 216-hour timeframe; (B) Changes in serum glucose levels over 48 hours and the AUC of serum glucose levels over a 48-hour timeframe. Figure 16 illustrates the body weight of db / db mice after administration of TE-8105 once daily sublingually for 5 days at different time points following administration. (A) Changes in body weight over time and AUC of body weight over a 216-hour timeframe; and (B) Changes in body weight over 72 hours and AUC of body weight over a 72-hour timeframe. Figure 17 illustrates the food and water intake of db / db mice after administration of TE-8105 once daily sublingually for 5 days at different time points following administration. (A) Changes in food intake over time and the AUC of food intake over a 216-hour period; and (B) Changes in water intake over time and the AUC of water intake over a 216-hour period. Figure 18 illustrates the serum glucose levels in db / db mice administered once daily sublingually at different pH values ​​for 5 days. (A) Changes in serum glucose levels over time and the AUC of serum glucose levels over a 192-hour timeframe; and (B) Changes in serum glucose levels over 120 hours and the AUC of serum glucose levels over a 120-hour timeframe; Figure 19 illustrates the body weight of db / db mice administered once daily sublingually at different pH values ​​for 5 days. (A) Changes in body weight over time and AUC of body weight over a 192-hour timeframe; and (B) Changes in body weight over 120 hours and AUC of body weight over a 120-hour timeframe; Figure 20 illustrates the food intake of db / db mice administered TE-8105 formulations at different pH values ​​once daily via sublingual administration for 5 days. (A) Changes in food intake over time and the AUC of food intake over a 192-hour period; and (B) Changes in food intake over 120 hours and the AUC of food intake over a 120-hour period. Figure 21 illustrates the water intake of db / db mice administered TE-8105 formulations at different pH values ​​once daily via sublingual administration for 5 days. (A) Changes in water intake over time and the AUC of water intake over a 192-hour period; and (B) Changes in water intake over 120 hours and the AUC of water intake over a 120-hour period. Figure 22 illustrates the serum glucose levels in db / db mice administered TE-8105 or smegglutinin twice daily sublingually for 3 days. (A) Changes in serum glucose levels over time; and (B) AUC of serum glucose levels over a 128-hour timeframe. Figure 23 illustrates serum glucose levels in db / db mice administered different concentrations of TE-8105 or smegglutinin twice daily via sublingual administration for 3 days. (A) Changes in serum glucose levels over time; and (B) AUC of serum glucose levels over a 128-hour timeframe. Figure 24 illustrates the serum glucose levels in db / db mice administered TE-8105 or smegglutinin sublingually once daily or every two days for 7 days. (A) Changes in serum glucose levels over time; and (B) AUC of serum glucose levels over a 271-hour time span. Figure 25 illustrates the body weight of db / db mice administered TE-8105 or smegglutinin sublingually once daily or every two days for 7 days. (A) Changes in body weight over time; and (B) AUC of body weight over a 264-hour timeframe; Figure 26 illustrates the food and water intake of db / db mice administered TE-8105 or smegglutinin sublingually once daily or every two days for 7 days. (A) Changes in food intake over a 264-hour period; and (B) Changes in water intake over a 264-hour period; Figure 27 illustrates the serum glucose levels in ZFDM rats treated with TE-8105 twice daily sublingually for 10 days. (A) Changes in serum glucose levels over time; and (B) AUC of serum glucose levels over an 18-day timeframe. Figure 28 illustrates the body weight of ZFDM rats treated with TE-8105 twice daily sublingually for 10 days. (A) Changes in body weight over time; and (B) AUC of body weight over an 18-day timeframe. Figure 29 illustrates the food intake of ZFDM rats treated with TE-8105 twice daily sublingually for 10 days. (A) Changes in food intake over time; and (B) AUC of food intake over an 18-day period. Figure 30 illustrates the water intake of ZFDM rats treated with TE-8105 twice daily via sublingual administration for 10 days. (A) Changes in water intake over time; and (B) AUC of water intake over an 18-day period; Figure 31 illustrates the serum glucose levels in db / db mice administered twice daily sublingually in liquid form containing peppermint oil and glycerin for 2 days. (A) Changes in serum glucose levels over time; and (B) AUC of serum glucose levels over a 120-hour timeframe. Figure 32 illustrates the serum glucose levels in db / db mice administered the TE-8105 formulation in gel form twice daily for 3 days. (A) Changes in serum glucose levels over time; and (B) AUC of serum glucose levels over a 168-hour timeframe. Figure 33 illustrates the body weight of db / db mice administered the TE-8105 formulation in soft gel form twice daily for 3 days. (A) Change in body weight over time; and (B) AUC of body weight over a 168-hour timeframe; Figure 34 illustrates the food and water intake of db / db mice administered the TE-8105 formulation in soft gel form twice daily for 3 days. (A) Changes in food intake over time and the AUC of food intake over a 168-hour period; and (B) Changes in water intake over time and the AUC of water intake over a 168-hour period. Figure 35 illustrates the serum glucose levels in db / db mice administered semaglutide in softgel form twice daily via sublingual administration for 3 days. (A) Changes in serum glucose levels over time; and (B) AUC of serum glucose levels over a 168-hour timeframe. Figure 36 illustrates the body weight of db / db mice administered the smegglutinin formulation in softgel form twice daily for 3 days. (A) Change in body weight over time; and (B) AUC of body weight over a 168-hour timeframe; Figure 37 illustrates the body weight of db / db mice administered the smegglutinin formulation in softgel form twice daily for 3 days. (A) Change in body weight over time; and (B) AUC of body weight over a 168-hour timeframe; Figure 38 illustrates the serum glucose levels in db / db mice treated twice daily with the TE-8105 formulation sublingually for 3 days. (A) Changes in serum glucose levels over time; and (B) AUC of serum glucose levels over a 168-hour timeframe. Figure 39 illustrates the body weight of db / db mice treated with TE-8105 tablets twice daily via sublingual administration for 3 days. (A) Change in body weight over time; and (B) AUC of body weight over a 168-hour timeframe. Figure 40 illustrates the food and water intake of db / db mice treated twice daily with TE-8105 tablets sublingually for 3 days. (A) Changes in food intake over a 168-hour period; and (B) Changes in water intake over a 168-hour period. Figure 41 illustrates the serum glucose levels in db / db mice treated twice daily with sublingual semaglutide tablets for 3 days. (A) Changes in serum glucose levels over time; and (B) AUC of serum glucose levels over a 168-hour timeframe. Figure 42 illustrates the body weight of db / db mice treated twice daily with sublingual semaglutide tablets for 3 days. (A) Changes in body weight over time; and (B) AUC of body weight over a 168-hour timeframe; Figure 43 illustrates the food and water intake of db / db mice treated twice daily with sublingual semaglutide tablets for 3 days. (A) Changes in food intake over a 168-hour period; and (B) Changes in water intake over a 168-hour period; Figure 44 illustrates the serum glucose levels in db / db mice treated twice daily with TE-8105 peppermint oil tablets sublingually for 3 days. (A) Changes in serum glucose levels over time; and (B) AUC of serum glucose levels over a 144-hour timeframe. Figure 45 illustrates the body weight of db / db mice treated with TE-8105 peppermint oil tablets twice daily via sublingual administration for 3 days. (A) Changes in body weight over time; and (B) AUC of body weight over a 144-hour timeframe; Figure 46 illustrates the food intake of db / db mice treated with TE-8105 peppermint oil tablets twice daily sublingually for 3 days. (A) Changes in food intake over time; and (B) AUC of food intake over a 144-hour period; and Figure 47 illustrates the water intake of db / db mice treated with TE-8105 peppermint oil tablets twice daily via sublingual administration for 3 days. (A) Changes in water intake over time; and (B) AUC of water intake over a 144-hour period.

[0045] Invention Description

[0046] The present disclosure provides a pharmaceutical formulation and its uses. Part of the present invention is developed based on the discovery that GLP-1 RA-P can be absorbed through the sublingual mucosal epithelium.

[0047] For convenience, specific technical terms used in this specification, examples, and the appended claims are grouped here. Unless otherwise defined in this specification, the meanings of scientific and technical terms used herein are the same as those understood and commonly used by those of ordinary skill in the technical field to which the present invention pertains.

[0048] Ranges of values are disclosed herein. The ranges set a lower limit value and an upper limit value. Unless otherwise defined in this specification, the ranges include all values up to the minimum value (lower limit value or upper limit value) and the ranges between the values of the range.

[0049] In this specification and the claims, the singular form "a" includes plural reference values, unless otherwise indicated by the context.

[0050] As used herein, the term "effective amount" refers to an amount sufficient to produce a therapeutic or prophylactic effect. According to an embodiment of the present disclosure, administering one or more doses of the formulation of the present disclosure to a diabetic individual (e.g., db / db mice) can reduce the blood glucose level of the individual to a normal value.

[0051] In this disclosure, "Bioavailability of GLP-1 RA-P" refers to the estimated bioavailability achieved by sublingual administration of GLP-1 RA-P relative to subcutaneous injection. Accordingly, "Bioavailability of GLP-1 RA-P" is expressed as a percentage of the bioavailability achieved by subcutaneous GLP-1 RA-P. To estimate the bioavailability of sublingual administration relative to subcutaneous injection in db / db mice, the hypoglycemic effects of two different doses of the sublingual administration were compared with a specific dose of a single subcutaneous injection at a time point of 24 hours after administration (before the third sublingual dose). At this time point, the subcutaneous injection resulted in low (normal) blood glucose levels. Both sublingual administrations, containing specific doses, also resulted in low blood glucose levels. The relative bioavailability was estimated by comparing the efficacy of the two sublingual administrations with the subcutaneous injection. For example, if two 200 nmol / kg sublingual administrations and a 50 nmol / kg subcutaneous injection have the same hypoglycemic effect at a 24-hour time point, the bioavailability of the sublingual administration relative to the subcutaneous injection can be estimated to be approximately 50 / 400 (or 12.5%). According to the preferred embodiments of this disclosure, the bioavailability of the GLP-1 RA-P disclosed herein (i.e., TE-8105, smegglutinin, or telpoxetine) delivered sublingually in db / db diabetic and obese mice has been observed to range from 7-10%.

[0052] In this document, the terms "individual," "patient," and similar terms may be used interchangeably, referring to mammals, preferably humans. An individual may be diagnosed with symptoms. In some cases, the symptoms are obesity. In some cases, the symptoms are type 2 diabetes. In some cases, the symptoms are a condition associated with obesity or type 2 diabetes. The individual may be male or female. The individual may be a patient of any age. Typically, the individual is a patient or other individual who is receiving a treatment regimen or is being evaluated for a treatment regimen (e.g., weight loss or treatment to lower hyperglycemia). However, in some embodiments, the individual is not receiving a treatment regimen.

[0053] 1. Pharmaceutical formulation of GLP-1 RA-P for sublingual administration

[0054] This disclosure relates to a non-injectable pharmaceutical formulation for the treatment of obesity, type II diabetes, or related conditions. The pharmaceutical formulation comprises GLP-1 RA-P as its active ingredient, and a pharmaceutically acceptable carrier, which may be a buffer, polymer, or the like, depending on the final form of the pharmaceutical formulation (e.g., liquid, tablet, colloid, etc.).

[0055] 1.1 This disclosure contains GLP-1 RA-P

[0056] Natural human GLP-1 is a peptide with approximately 30 amino acid residues and an extremely short half-life of only about 2-5 minutes in humans. GLP-1 RA-Ps that have been approved or developed for clinical use generally have amino acid substitutions to minimize their sensitivity to serum proteases (e.g., dipeptidyl peptidase-4 (DPP-4)) and thus prolong their half-life in serum. Examples of GLP-1 RA-Ps applicable to this disclosure, according to embodiments thereof, include, but are not limited to, TE-8105 (as illustrated in Figure 1), semaglutide, and telpolide. TE-8105 can be prepared according to the process described in Example 1 of this disclosure or Example 2 of US Patent No. US2020 / 022498A1; semaglutide and telpolide are available from commercial sources. These GLP-1 RA-Ps all share a common structural feature: they are all modified and contain long-chain fatty acids with conjugation linkages. Smegglutinin and TE-8105 contain a peptide motif composed of 31 amino acid residues, derived from a fragment of natural human GLP-1 (residues #7-37), with artificially introduced amino acid substitutions to resist degradation by serum protease (DPP-4). In both cases, the 20th amino acid residue is a lysine residue, whose ε-amino group is conjugated to the long-chain C-18 diacid in smegglutinin and the fatty acid bundles of the C16 and C18 diacids in TE-8105. Conversely, telpolide is a peptide composed of 39 amino acid residues, designed to be homologous to human GLP-1 and human glucose-dependent peptide (GIP). The 20th residue of telpolide is also a lysine residue, which is conjugated to a long-chain C20 diacid. Modifying GLP-1 RA-P with long-chain fatty acids can produce the following physiological benefits: Strengthening the binding with serum albumin: Once GLP-1 RA-P crosses the sublingual mucosal epithelium, it reaches the lamina propria and diffuses into the bloodstream. Enhancing the binding of GLP-1 RA-P with albumin in the interstitial spaces slows its diffusion into the bloodstream, and the circulating albumin-bound form also prolongs its serum half-life, thereby improving bioavailability.

[0057] It improves the affinity of peptides for mucosal epithelial cell membranes, including components such as lipids and proteins, thus accelerating the passage of these peptides through epithelial cells.

[0058] 1.2 Sublingual administration formulation

[0059] The sublingual region in humans is relatively small and has limited space. Therefore, this disclosure aims to provide a sublingual administration formulation containing an optimal amount of GLP-1 RA-P within a maximum pharmaceutically acceptable carrier. Upon administration, compared to subcutaneous injection, the pharmaceutical formulation of this disclosure can relatively improve the bioavailability of GLP-1 RA-P, thereby producing a therapeutic effect.

[0060] (i) Liquid formulation

[0061] Designing liquid sublingual drug delivery formulations requires consideration of numerous factors to achieve the highest bioavailability of drug molecules and thus produce the desired therapeutic effect. These factors include, at a minimum, the maximum amount of drug molecules that can be absorbed by a single sublingual dose, the maximum volume of a single dose, and the maximum amount of drug that can be carried in a single dose (i.e., drug solubility).

[0062] According to embodiments of this disclosure, the maximum amount of GLP-1 RA-P that can be absorbed or transported across mucosal cell membranes, TE-8105, is approximately 3-5 mg / mL, and smegglutinin is 3.2 mg / mL. These maximum amounts are affected by the pH of the buffer solution. According to other embodiments of this disclosure, a suitable volume for a single sublingual administration is approximately 1.5-2.0 mL. In this case, when the volume is greater than 2.0 mL, the drug will spill from the sublingual region into other spaces in the oral cavity, and the drug components may enter the gastrointestinal tract with swallowing. Accordingly, the liquid formulation of this disclosure is designed to contain 5-10 mg of GLP-1 RA-P dissolved in 1.5-2.0 mL of buffer solution with a pH between 5.5 and 7.5.

[0063] According to a preferred embodiment of this disclosure, the pharmaceutical formulation is prepared as follows: In deionized water, a buffer (e.g., histidine), a specific amount of GLP-1 RA-P (i.e., TE-8105, smegglutide, or telpolide), and an osmotic pressure regulator (e.g., mannitol) are mixed until the GLP-1 RA-P is completely dissolved. The solution is then adjusted to a desired pH, preferably between 5.5 and 7.5. For example, 5-10 mg of GLP-1 RA-P (such as 5, 6, 7, 8, 9, or 10 mg of GLP-1 RA-P) can be dissolved in 1.5-2.0 mL of buffer solution, for example, 1.5, 1.6, 1.7, 1.8, 1.9, or 2.0 mL of buffer solution, adjusting the pH to between 5.5 and 7.5, for example, 5.5, 5.6, 5.7, 5.8, 5.9, 6.0, 6.1, 6.2, 6.3, 6.4, 6.5, 6.6, 6.7, 6.8, 6.9, 7.0, 7.1, 7.2, 7.3, 7.4, or 7.5. In some embodiments, GLP-1 RA-P is TE-8105, and the formulations disclosed herein contain 5-10 mg of TE-8105 dissolved in 1.5-2.0 mL of buffer solution. In other embodiments, GLP-1 RA-P is semaglutide, and the formulation disclosed herein contains 5-6 mg of semaglutide dissolved in 1.5-2.0 mL of buffer solution. In still other embodiments, GLP-1 RA-P is telpolide, and the formulation disclosed herein contains 5-10 mg of telpolide dissolved in 1.5-2.0 mL of buffer solution.

[0064] Alternatively or alternatively, in addition to the aforementioned buffer solution, any solution having an osmotic pressure equivalent to that of the human body can be used to prepare the liquid formulation of this disclosure, such as phosphate-buffered saline (PBS). Alternatively or additionally, preservatives (e.g., phenol), flavoring agents (e.g., peppermint oil), and / or emollients (e.g., glycerin) may be added to the aforementioned liquid formulation. According to certain embodiments of this disclosure, the liquid formulation further comprises 0.1-0.5% (w / v) peppermint oil, for example: 0.1, 0.2, 0.3, 0.4, or 0.5% (w / v) peppermint oil.

[0065] (ii) Soft rubber formulation

[0066] Optionally or additionally, the aforementioned liquid formulation may be formulated with gelatin to produce a soft gel formulation that melts within 1-2 minutes when delivered to the sublingual space.

[0067] According to embodiments of this disclosure, the liquid formulation is mixed with 5-10% (w / v) gelatin, for example, with 5, 6, 7, 8, 9, or 10% (w / v) gelatin to form a mixture, and forms a soft gel at a low temperature (e.g., 4°C) that can be easily picked up with tweezers. The gelatin suitable for this disclosure can be derived from cold-water fish species. In some embodiments, the liquid formulation of this disclosure is mixed with 5% (w / v) cold-water fish gelatin. In other embodiments, the liquid formulation of this disclosure is mixed with 10% (w / v) cold-water fish gelatin.

[0068] (iii) Tablet Formulation

[0069] Alternatively or additionally, the aforementioned liquid formulation may be formulated together with the disintegrating polymer into tablets that disintegrate immediately upon contact with water or saliva, releasing the drug molecules contained in the tablets.

[0070] According to this disclosure, the liquid formulation is mixed with 1-10% (w / v) of cross-linked povidone, such as 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10% (w / v) of cross-linked povidone, and after drying, it forms a tablet that can be easily picked up with tweezers and delivered to the sublingual area of ​​an individual.

[0071] 2. Treatment methods

[0072] The pharmaceutical formulations described in Part 1 of this specification are intended for the treatment of diseases or conditions related to hyperglycemia. Accordingly, another aspect of this disclosure aims to provide a method for treating obesity, type II diabetes, or related diseases. The method comprises: An effective amount of a pharmaceutical formulation is administered to an individual via sublingual administration and the pharmaceutical formulation is maintained under the individual's tongue for approximately 3-10 minutes, wherein the pharmaceutical formulation contains GLP-1 RA-P. in The GLP-1 RA-P is selected from the group consisting of TE-8105, smegglutinin and telpokinin; The pH value of the pharmaceutical formulation is between 5.5 and 7.5; Pharmaceutical formulations are administered once daily, twice daily, every other day, once weekly, or twice weekly; and Compared to administration to individuals via subcutaneous injection, the pharmaceutical formulation achieves a bioavailability of 2-10% for GLP-1 RA-P.

[0073] According to embodiments of this disclosure, the pharmaceutical formulation may be in liquid, tablet, or soft gel form and may be maintained under the tongue for 3-10 minutes, for example: 3, 4, 5, 6, 7, 8, 9, or 10 minutes; preferably 5-10 minutes, for example: 5, 6, 7, 8, 9, or 10 minutes. In some cases, the pharmaceutical formulation is maintained under the tongue for 3 minutes. In some cases, the pharmaceutical formulation is maintained under the tongue for 5 minutes. In some cases, the pharmaceutical formulation is maintained under the tongue for 7 minutes. In some cases, the pharmaceutical formulation is maintained under the tongue for 10 minutes. In all embodiments of this disclosure, the individual's blood glucose level, body weight, and food and water intake may decrease to low normal levels after administration of the pharmaceutical formulation.

[0074] In some embodiments, the pharmaceutical formulation is the aforementioned liquid formulation and comprises 5-10 mg of GLP-1RA-P dissolved in 1.5-2.0 mL of buffer solution. In some embodiments, the liquid formulation comprises 5-10 mg of TE-8105 dissolved in 1.5-2.0 mL of buffer solution, administered once daily, twice daily, every two days, once weekly, or twice weekly. In some embodiments, the liquid formulation comprises 5-6 mg of smegglutide dissolved in 1.5-2.0 mL of buffer solution, administered twice daily or every two days. In some embodiments, the liquid formulation comprises 1-10 mg of telpolide dissolved in 1.5-2.0 mL of buffer solution, administered twice daily.

[0075] In some embodiments, the pharmaceutical formulation is the aforementioned soft gel formulation, prepared by mixing the liquid formulation disclosed herein with 5-10% (w / v) gelatin. In some embodiments, the soft gel formulation is prepared by mixing the TE-8105 liquid formulation with 5% or 10% (w / v) gelatin and administered at the same administration frequency as the liquid formulation. In some embodiments, the soft gel formulation is prepared by mixing the smegglutinin liquid formulation with 5% or 10% (w / v) gelatin and administered at the same administration frequency as the liquid formulation.

[0076] In some embodiments, the pharmaceutical formulation is the aforementioned tablet formulation, prepared by mixing the liquid formulation disclosed herein with 1-10% (w / v) of crospovidone. In some embodiments, the tablet formulation is prepared by mixing the TE-8105 liquid formulation with 1% (w / v) of crospovidone and administered at the same administration frequency as the liquid formulation. In some embodiments, the tablet formulation is prepared by mixing the smegglutide liquid formulation with 1% (w / v) of crospovidone and administered at the same administration frequency as the liquid formulation.

[0077] According to certain embodiments of this disclosure, as demonstrated in diabetic and obese db / db mouse models, the bioavailability of sublingual TE-8105 is 7-8% that of subcutaneous injection. According to certain embodiments of this disclosure, the bioavailability of sublingual semaglutide is 7% that of subcutaneous injection. According to other embodiments of this disclosure, the bioavailability of sublingual semaglutide is 10% that of subcutaneous injection. Compared to two known and approved peptide hormone drugs for sublingual administration (oxytocin and desmopressin), whose bioavailability is less than 1%, the achievement of a 7-10% bioavailability by the pharmaceutical formulations and / or methods of this disclosure is an unexpected finding.

[0078] According to the embodiments disclosed herein, the diseases related to obesity or type II diabetes that can be treated by the methods disclosed herein are overweight, fatty liver, non-alcoholic fatty liver disease, diabetic cardiomyopathy, or coronary atherosclerotic heart disease.

[0079] Example

[0080] Materials and Methods

[0081] Preparation of TE-8105, smegglutinin and telposide samples for use in this disclosure

[0082] The TE-8105 sample was prepared from a GMP-grade pharmaceutical ingredient manufactured by CDMO Shanghai Hequan Pharmaceutical Co., Ltd. (Shanghai, China) for Phase I and Phase II human clinical trials. The pharmaceutical ingredient was initially in powder form and was reconstituted into a liquid with TE-8105 at a concentration of 5 mg / mL. The excipients used included histidine, phenol, mannitol, hydrochloric acid, and sodium chloride at pH 7.0.

[0083] In some trials, a liquid formulation of the drug, manufactured by CDMD, at a concentration of 5 mg / mL, was used, containing the same excipients as listed above. Additionally, Ozempic, a commercially available product suitable for clinical use, was purchased from a commercial source, with each 1.5 mL vial containing 2 mg of the drug (1.34 mg / mL). Ozempic formulation contains excipients such as disodium hydrogen phosphate dihydrate, propylene glycol, phenol, hydrochloric acid, sodium hydroxide (for pH adjustment), and water for injection, and has a final pH of 7.0.

[0084] The telpolide powder (batch No. V423401) used for research purposes was purchased from Invivo Chem (Libertyville, Illinois, USA). In this study, the telpolide powder was dissolved in a liquid formulation to a concentration of 5 mg / mL, containing the excipient disodium hydrogen phosphate heptahydrate, sodium chloride, concentrated hydrochloric acid, sodium hydroxide (to adjust pH), and water for injection. These excipients are the same as those used in the clinical product Mounjaro.

[0085] For each GLP-1 RA-P, it was diluted with an individual liquid formulation as a dilution buffer to prepare different doses.

[0086] Using the db / db mouse model to study sublingual delivery of GLP-1 RA-P and its bioavailability

[0087] db / db mice are inbred strains of mice carrying a homologous mutation in the leptin receptor gene, resulting in voracious eating behavior and obesity, typically reaching twice the normal body weight. These mice typically exhibit serum glucose levels that rise from below 200 mg / mL to above 400 mg / mL, and HbA1c levels that increase from below 6% to above 8%, or higher. These parameters are very similar to those of patients with type 2 diabetes and obesity. Furthermore, serum glucose levels in db / db mice are highly sensitive to changes in GLP-1 RA-P drug concentrations, making glucose measurement a valid indicator of drug availability in the blood. In the studies described in this specification, the db / db mouse model was used to measure the efficiency of sublingual administration, estimating bioavailability through sublingual and subcutaneous injections, and detecting the cumulative effect of repeated sublingual administration.

[0088] These experiments used 30 male db / db mice, aged 6 weeks and weighing between 35 and 40 grams. After a 2-week acclimatization period in the laboratory, the mice's weight and blood glucose levels were measured. Mice with blood glucose levels between 300 and 400 mg / mL were selected for the experiments. After receiving GLP-1 RA-P treatment, the mice were allowed to rest for at least one week before subsequent testing. When the mice matured to 20-25 weeks of age, their weight typically increased by 50-60 grams.

[0089] GLP-1 RA-P was administered sublingually using the following method: Mice were anesthetized with isoflurane vapor (2.5–3%, 700–800 mL / min) by placing a tube near the nose for 6–7 minutes. The mice were placed dorsally on a flexible polystyrene foam board. To expose the sublingual area, a leash was looped around the lower incisors, and the jaw was gently pulled down and secured to the foam board with tape. The liquid formulation of GLP-1 RA-P (20–50 μL) was slowly applied to the sublingual area over 5 minutes. Once the anesthesia wore off, the mice quickly regained their mobility. During the process, all or part of the GLP-1 RA-P was absorbed by the sublingual epithelium.

[0090] Using a Zucker rat model of fatty-fatty diabetes (ZFDM) to study sublingual delivery of GLP-1 RA-P and its bioavailability usage rate

[0091] This study investigated the feasibility of sublingual administration of TE-8105 in ZFDM rats, and its efficacy in glucose control and weight reduction. ZFDM rats are a subline with a missense mutation in the leptin receptor gene. These rats develop obesity between the first 6 and 12 weeks of age and diabetes between 10 and 20 weeks. Their weight can reach over 400 grams, and their fasting blood glucose levels do not rise to 400-500 mg / dL. Therefore, these rats serve as a valuable model for studying potential treatments for obesity and diabetes.

[0092] In the experiments disclosed herein, the ZFDM (fa / fa) rats used were 8-10 weeks old, weighed approximately 150-200 grams, and had a non-fasting blood glucose level of 120-150 mg / dL. The sublingual delivery procedure for GLP-1 RA-P was performed according to the procedure described above for db / db mice.

[0093] Example 1: Synthesis of TE-8105

[0094] TE-8105 is synthesized by combining two structural moieties via a copper(I)-catalyzed olefin-azide click reaction, wherein the structures of moieties I and II are as follows: Part I: GLP-1 agonists containing 2-amino-isobutyric acid (Aib) substituted with azidoides

[0095] Part II: Alkyne-2FA-C16-acid-C18-diacid

[0096] 1.1 Synthesis of GLP-1 agonists substituted with Aib containing azide

[0097] Part I of TE-8105 was synthesized according to the process described by Zhang et al. (US 2020 / 022498A1). In short, the peptide moiety (Sequence No. 1) was first synthesized using solid-phase peptide synthesis (SPPS), followed by the substitution of the alanine residue at position 2 of Sequence No. 1 with a 2-aminoisobutyric acid residue to produce an Aib-substituted GLP-1 agonist (Sequence No. 2). Notably, the Aib residue is included in the GLP-1 agonist to ensure the molecule's resistance to DPP-4 degradation. Next, the γ-carboxyl group of the glutamate residue is linked to the ε-amino group of the lysine residue in the Aib-substituted GLP-1 agonist molecule (Sequence No. 2). Finally, the α-amino group of the glutamate residue is modified with an azidoacetyl group to produce an Aib-substituted GLP-1 agonist molecule containing an azide (or Part I of TE-8105).

[0098] The peptide synthesis procedure comprises Fmoc SPPS performed step-by-step using an O-benzotriazole-N,N,N',N'-tetramethylurea hexafluorophosphate (HBTU) / N,N-diisopropylethylamine (DIEA) / N,N-dimethylformamide (DMF) coupling chemistry, wherein HBTU acts as an in-situ activator for the Fmoc-protected amino acid, and DIEA acts as an organic base in the coupling process. Nα-Fmoc is a side-chain protected amino acid, and 2-chlorotriacyl chloride resin (CTC resin) is used in the synthesis. Side-chain protection strategies are used for the following Fmoc-protected amino acids: Fmoc-Arg (Pbf), Fmoc-Trp (Boc), Fmoc-Thr (OtBu), Fmoc-Lys (N-Dde), Fmoc-Tyr (OtBu), Fmoc-Glu (OtBu), Fmoc-Gln (Trt), Fmoc-Ser (OtBu), and Fmoc-His (Trt). Other Fmoc-protected amino acids used in the synthesis of GLP-1 agonists include: Fmoc-Gly-OH, Fmoc-Val-OH, Fmoc-Leu-OH, Fmoc-Ala-OH, Fmoc-Ile-OH, Fmoc-Phe-OH, and Fmoc-Aib-OH. Abbreviations: Pbf, 2,2,4,6,7-pentamethyldihydrobenzofuran-5-sulfonyl chloride; Boc, tert-butyloxycarbonyl; tBu, tert-butyl ether; Dde, 1-(4,4-dimethyl-2,6-dioxocyclohexyl-1-ethylene)ethyl; Trt, triphenylmethyl; TIS, triisopropylsilane; Fmoc, 9-fluorenylmethoxycarbonyl.

[0099] In each coupling cycle except the first coupling cycle, the system uses 3 nmol of N.α The Fmoc protecting group on the α-amine was removed by using 6 nmol of DIEA and 2.85 nmol of the equivalent HBTU with a solution of 20% piperidine in dimethylformamide (DMF) three times the volume of the peptide resin.

[0100] Step (i): Initial Coupling

[0101] Peptide synthesis began with the covalent linking of the first amino acid to the resin. Fmoc-Gly-OH (1.0 mmol, 297.5 mg) and CTC resin (1.0 mmol, substitution = 1.0 mmol / g, 1.0 g) were dissolved in dichloromethane (DCM), followed by the addition of DIEA (4.0 nmol). The mixture was stirred at 20°C for 2 hours under nitrogen atmosphere. Methanol (MeOH, 1.0 mL) was added as a capping agent to react with the unreacted carbocations on the CTC resin, and the mixture was stirred at 20°C.

[0102] Step (ii): Washing

[0103] Remove the capping agent solution containing methanol and wash the resin three times with DMF.

[0104] Step (iii): Fmoc deprotection

[0105] The Fmoc protecting groups on the CTC resin were removed by treating it with a 20% piperidine DMF solution at 20°C for 0.5 hours.

[0106] Step (iv): Washing

[0107] After deprotection, the solution was removed and the resin was washed five times with DMF.

[0108] Step (v): Coupling the second amino acid

[0109] The second amino acid (tri-equivalent Fmoc-Arg(pbf))-OH) and the activator HBTU were added to the resin. The mixture was stirred under nitrogen for 1 hour. Steps (ii) to (v) were repeated sequentially for each amino acid in the peptide sequence (sequence number: 1). Each coupling reaction was monitored using the Ninghai standard assay.

[0110] Cutting and purification

[0111] To cleave the side-chain protected peptides from CTC resin, a cleavage buffer (40.0 mL; 2.5% TIS / 2.5% H2O / 2.5% 3-mercaptopropionic acid / 92.5% trifluoroacetic acid (TFA)) was prepared and added to a flask containing the resin. The mixture was stirred at room temperature for 2 hours. Cold isopropyl ether (500.0 mL) was added and the mixture was centrifuged at 6000 rpm for 3 minutes to precipitate the crude peptides. The crude peptides were then washed twice with isopropyl ether and dried under vacuum for 2 hours.

[0112] Aib-substituted GLP-1 agonists (or TE-8105 fraction I) containing azide were purified by reversed-phase HPLC on Gemini C18 columns (5 μm, 100 Å) and Luna columns (10 μm, 100 Å). The mobile phase A was water (containing 0.075% trifluoroacetic acid), and the mobile phase B was acetonitrile. A linear gradient of 17%–47% acetonitrile was applied over 60 minutes at a flow rate of 20 mL / min, and the column temperature was maintained at 25 °C. The HPLC chromatogram showed a residence time of 11.65 min for the major peak at OD254, indicating the presence of an Aib-substituted GLP-1 agonist containing azide.

[0113] The Aib-substituted GLP-1 agonist containing an azide (or part I of TE-8105) was purified by reversed-phase HPLC using a Gemini C18 column (5 μm, 100 Å) and Luna (10 μm, 100 Å). The mobile phase consisted of A: water containing 0.075% trifluoroacetic acid and B: acetonitrile. A linear gradient of 17%–47% acetonitrile was applied over 60 minutes at a flow rate of 20 mL / min, and the column temperature was maintained at 25 °C. The HPLC chromatograms showed that the major peak of the Aib-substituted GLP-1 agonist containing an azide was located at OD254, with a residence time of 11.65 min, confirming the identity of the molecule.

[0114] 1.2 Synthesis of alkyne-2FA-C16-acid-C18-diacid

[0115] In this embodiment, part II of the preparation of TE-8105, namely, the preparation of a linker unit containing alkyne, is characterized by having palmitoleic acid and octadecanoic acid (represented as "alkyn-2FA-C16-acid-C18-diacid"). The peptide core is alkynyl-ethyl-Xaa4-K(C16)-Xaa4-K(C18-acid)-OH, containing two lysine (K) residues and an alkynyl propionyl group at the N-terminus. The spacers between the two lysines, and between the alkynyl propionyl group and the adjacent lysine, are PEGylated amino acids (represented as "Xaa4") having four ethylene glycol repeating sequences. Two fatty acid chains with different lengths and terminal functional groups (C16-acid and C18-diacid) conjugate to the lysine residues by forming amide bonds between the carboxyl group (-CO2H) of the fatty acid and the amino group of the lysine. The novel molecule, alkyne-ethyl-Xaa4-K(C16)-Xaa4-K(C18-acid)-OH, was synthesized according to the following steps.

[0116] Step (i): Coupling the first amino acid

[0117] The synthesis step begins with the conjugation of the first amino acid to the resin. Fmoc-Lys(Dde)-OH (2.0 mmol, 1 equivalent) and CTC resin (2.0 mmol, 1 equivalent) are dissolved in dichloromethane (DCM, 10.0 mL). DIEA (8.0 mmol, 4 equivalent) is added to the mixture, and the mixture is expanded in nitrogen bubbles at 20 °C for 2 hours.

[0118] Step (ii): Washing

[0119] The peptide-resin mixture was drained and washed three times with DMK.

[0120] Step (iii): Deprotection of the Fmoc base

[0121] Treatment with a DMF solution containing 20% ​​piperidine for 3 hours was performed to remove the Fmoc protecting group.

[0122] Step (iv): Washing

[0123] Drain the resin and wash it five times with DMF.

[0124] Step (v): Coupling of Fmoc-Glu-OtBu

[0125] Fmoc-Glu-OtBu (4.0 mmol, 2 equivalents) was coupled by adding DIEA (8.0 mmol, 4 equivalents) and HATU (3.8 mmol, 1.9 equivalents) to the resin and expanding it in nitrogen bubbles at 20°C for 30 minutes. The product was washed five times with DMF.

[0126] Step (vi): Repeat step (vi) for subsequent coupling.

[0127] Repeat steps (ii) through (vi) to couple 8-(tert-tert-butyl)-18-oxadianoic acid (2 equivalents). After coupling, add 3% hydrazine / DMF solution to the peptide-resin mixture and treat for 15 minutes. Wash the product five times with 20 mL of DMF.

[0128] Coupling of other components

[0129] Repeat steps (ii) through (vi) for each of the following components: Fmoc-NH-PEG4-C H (1.5 equivalent) Fmoc-Lys(Dde)-OH (2.0 equivalents) Fmoc-NH-PEG4-C H (1.5 equivalent) 4-Pentyn-1-acid (2.0 equivalents) After completing the coupling of 4-pentyne-1-acid, treat with 3% hydrogen nitrogen / DMF for 20 minutes. Wash the product with DMF five times. Repeat steps (ii) to (vi) to couple FMOC-GLU-OTBU (2.0 equivalents) and palmitic acid (2.0 equivalents).

[0130] Cutting and purification

[0131] Side-chain protected peptides were cleaved from CTC resin using a cleavage buffer (1% TFA / 99% DCM), which was added to the peptide resin at room temperature and treated twice for 5 minutes. The crude peptides were concentrated under pressure and lyophilized to yield 1.27 g of a yellow solid powder. The synthesis was confirmed by liquid chromatography-mass spectrometry (LC-MS). The residence time of the synthesized molecule containing an alkyne linker with two aliphatic chains was 2.769 min, confirming the expected molecule.

[0132] 1.3 Synthesis of TE-8105 (or 2FA-GLP-1 receptor agonist)

[0133] In this embodiment, the GLP-1 receptor agonist described for synthesis refers to "2FA-GLP-1 receptor agonist" or TE-8105. TE-8105 is synthesized by coupling the Aib-substituted azide-containing GLP-1 agonist of Example 1.1, which contains an azide, to the alkyne-containing linker unit of Example 1.2, which contains a palmitoleic acid and an octadecanoic acid, through a copper-catalyzed azide-alkyne cycloaddition reaction (CuAAC) between the azide and alkyne functional groups.

[0134] In summary, a mixture of an Aib-substituted GLP-1 agonist (1.22 equivalents, TFA) containing an azide and an alkyne linker (1 equivalent, TFA) was prepared in DMF (4 mL). The solution was degassed and purged three times with nitrogen. Copper(I) (1 equivalent) and DIEA (4.0 equivalents) were added to the reaction mixture, followed by stirring at 20 °C under nitrogen for 10 minutes. The reaction was confirmed by LC-MS, where the residence time of the 2FA-GLP-1 receptor agonist (TE-8105) was observed to be 1.784 minutes.

[0135] The 2FA-GLP-1 receptor agonist was purified sequentially by reversed-phase HPLC on Luna C18 columns (10 μm, 100 Å) and Gemini C18 columns (5 μm, 100 Å). Mobile phase A, containing 0.075% trifluoroacetic acid, and mobile phase B, containing acetonitrile, were used for purification. A linear gradient of 30% to 60% acetonitrile was applied over 60 minutes at a flow rate of 20 mL / min, while the column temperature was maintained at 30°C. The HPLC chromatogram of the purified product showed a major peak at OD214 / OD254 nm with a residence time of 12.587 min, confirming the formation of TE-8105. The purified product was then lyophilized, yielding 18.3 g of a white powder (yield 10.1%, purity 96.4%, TFA).

[0136] Example 2: Bioavailability, additional and sustained efficacy of TE-8105 to db / db mice after repeated sublingual administration for 3 days via twice-daily (2QD) administration.

[0137] In this embodiment, the bioavailability of TE-8105 and its additive and sustained effects on serum glucose levels were assessed using the db / db mouse model described in the "Materials and Methods" section. To achieve this, the drug was administered sublingually (SL) twice daily (7 hours apart, or twice daily, 2 QD) for three consecutive days. Blood samples were collected before each TE-8105 administration, and serum glucose levels were measured. The results are illustrated in Figures 2, 3, and 4.

[0138] The results showed that blood glucose levels could be reduced to normal, and continuous sublingual administration of TE-8105 maintained normal blood glucose concentrations; when administration ceased, blood glucose levels rose to abnormal levels within 2-3 days (Figure 2A). Furthermore, blood glucose levels at the first three time points after sublingual administration of different doses were compared with those in the subcutaneous injection group (2 doses of 200 nmol / kg, total 400 nmol / kg) (Figure 2B). With a total sublingual administration of 400 nmol / kg, the blood glucose-reducing effect was similar to that of the subcutaneous injection group (34 hours) at 24 hours. Therefore, the bioavailability of sublingual TE-8105 is approximately 7-8% (30 / 400) of the bioavailability achievable with subcutaneous TE-8105.

[0139] The results showed the area under the curve (AUC) of serum glucose over 168 hours, comparing the efficacy of six doses of sublingual administration versus a single subcutaneous injection in lowering serum glucose levels. The results indicated that continuous sublingual administration at doses of 100 nmol / kg and 200 nmol / kg achieved better hypoglycemic effects than subcutaneous injection.

[0140] Statistical analysis revealed variability in blood glucose measurements at different time points. This variability arose from endogeneous differences among individual mice or from the bioassay itself. This conclusion is supported by smooth and consistent patterns of blood glucose variation observed in three independent mice across each group (Figure 3).

[0141] The additional and sustained effects of repeated sublingual administration of different doses of TE-8105 to db / db mice are illustrated in Figure 4. Figure 4A compares the efficacy of six sublingual doses of TE-8105 (100 nmol / kg, 150 nmol / kg, and 250 nmol / kg) administered over two QD, versus a single subcutaneous injection of 30 nmol / kg TE-8105 in reducing serum glucose levels. Figure 4B illustrates the effect after two sublingual administrations, with the third glucose time point obtained before the third administration. Figure 4C illustrates the dose-dependent effect on AUC.

[0142] Example 3: Bioavailability, additional and sustained efficacy of smegglutinin administered sublingually for 3 days in db / db mice via 2QD

[0143] In this embodiment, the bioavailability of smegglutinin and its additive and sustained effects on serum glucose levels were assessed using a db / db mouse model following a similar procedure to that described in Example 2. Smegglutinin was administered twice daily for three consecutive days, with a 7-hour interval between each administration. Blood samples were collected prior to each administration to measure serum glucose levels. The results are illustrated in Figure 5.

[0144] The results showed that continuous sublingual administration of semaglutide effectively reduced and maintained serum glucose levels within normal ranges. However, if administration was interrupted, serum glucose levels rapidly returned to abnormal levels within 1-2 days (Figure 5A). Furthermore, serum glucose levels at the first three time points after sublingual administration of different doses of semaglutide were compared with those achieved through subcutaneous injection (Figure 5B). The results showed that two sublingual doses of 350 nmol / kg (total dose 700 nmol / kg) had a similar glucose-lowering effect at 24 hours (the third time point) as a single subcutaneous injection of 50 nmol / kg. Therefore, the bioavailability of sublingual semaglutide was approximately 7% (50 / 700) of the subcutaneous formulation. The AUC values ​​shown in Figure 5C indicate the dose-dependent effect of sublingual semaglutide in lowering serum glucose.

[0145] Example 4: Bioavailability, additional and sustained efficacy of telpopritide administered sublingually for 3 days in db / db mice via 2QD

[0146] In this embodiment, the bioavailability of telperidine and its additive and sustained effects on serum glucose levels were assessed using a db / db mouse model following a similar procedure to that described in Example 2. Telperidine was administered twice daily for three consecutive days, with a 7-hour interval between each administration. Blood samples were collected prior to each administration to measure serum glucose levels. The results are illustrated in Figure 6.

[0147] As illustrated in Figure 6A, serum glucose levels can be reduced to normal, and continued administration of telpotetin can maintain serum glucose levels at normal levels; however, if administration is interrupted, serum glucose levels will rapidly return to abnormal levels within 1-2 days. Serum glucose levels at the first 3 time points after sublingual administration of different doses of telpotetin were compared with those achieved by subcutaneous injection. The results showed that sublingual administration of two doses of 500 nmol / kg (total dose 1000 nmol / kg) of telpotetin had a similar glucose-lowering effect on the 24-hour time point (3rd time point) as a single subcutaneous injection of 100 nmol / kg of telpotetin. Therefore, the bioavailability of sublingual telpotetin is approximately 10% (100 / 1000) of that of subcutaneous telpotetin. The AUC values ​​illustrated in Figure 6C indicate the dose-dependent effect of sublingual telpotetin in lowering serum glucose.

[0148] Example 5: Comparison of the effects of twice-daily sublingual or oral administration of TE-8105 to db / db mice for three days on serum glucose, body weight, and food and water intake.

[0149] In this embodiment, TE-8105 was administered sublingually (SL) or orally (PO) twice daily for three days, and the mice's blood glucose levels, body weight, and food and water intake were measured. For sublingual administration, mice were anesthetized and the liquid formulation of TE-8105 was maintained in the sublingual region for 5-10 minutes. For oral delivery, mice were kept upright and a liquid formulation of the same amount of TE-8105 but a different concentration was delivered into the mouth using a micropipette over 1-2 minutes; the mice quickly swallowed the liquid. The results are summarized in Figures 7-10.

[0150] The results in Figure 7 show that oral administration of TE-8105 did not reduce blood glucose levels in db / db mice, while sublingual administration effectively reduced blood glucose, with larger volumes of the same amount showing better results. Regarding body weight, oral administration of TE-8105 did not reduce body weight in db / db mice; however, sublingual administration of 200 nmol / kg and 1.4 mg / ml of TE-8105 significantly reduced body weight (Figure 8). Furthermore, the pattern of food intake reduction and body weight reduction in db / db mice via different routes (SL or PO) was very similar to that of body weight reduction (Figure 9). As for the effect on water intake, oral administration of TE-8105 did not affect water intake, while sublingual or subcutaneous administration reduced water intake (Figure 10).

[0151] Example 6: Comparison of the effects of sublingual administration of different concentrations of TE-8105 to db / db mice once daily for five days on serum glucose, body weight, and food and water intake.

[0152] In this embodiment, the concentrations and volumes of different liquid formulations of TE-8105 were tested. The sublingual regions of animals and humans are quite similar. To maximize the amount of drug transported across the sublingual mucosa, it is best to utilize as much of the sublingual mucosa as possible. Therefore, the volume of the liquid formulation is a relevant factor. Furthermore, transmucosal transport must involve the interaction between drug molecules in the liquid formulation and the surface components of the mucosal epithelial cells. Therefore, there must be an optimal concentration of drug molecules. In this embodiment, the same dose of TE-8105 (200 nmol / kg) was prepared into three concentrations (1.4, 3, and 5 mg / ml), and their absorption rates and subsequent pharmacokinetic effects were compared. The results are summarized in Figures 11 to 14.

[0153] The results showed that the liquid formulation of TE-8105 effectively reduced blood glucose at all three concentrations, with the 3 mg / ml concentration exhibiting the best efficacy (Figure 11). The kinetic pattern for the first three days is shown in Figure 9(B), which showed almost the same trend as the long-term pattern. Regarding weight reduction, all concentrations of the liquid formulation of TE-8105 demonstrated weight-reducing efficacy in obese db / db mice, with the 3 mg / ml concentration showing the best efficacy (Figure 12). In addition, all three concentrations of TE-8105 reduced food and water intake in db / db mice; however, there was no statistically significant difference among the three concentrations of TE-8105 (Figures 13 and 14).

[0154] Example 7: Comparison of the effects of once-daily sublingual administration of TE-8105 mice at different times and durations for five days on serum glucose, body weight, and food and water intake.

[0155] In typical practice, patients are instructed to remain still in the tongue and mouth for 5-10 minutes when taking medication sublingually to allow sufficient time for the drug to contact and be absorbed by the mucosal epithelial cells. Accordingly, this example investigates the efficiency of TE-8105 in penetrating the sublingual epithelium of db / db mice, where the TE-8105 liquid formulation was placed under the tongue of anesthetized mice for 1, 3, 5, 7, and 10 minutes before the mice recovered from anesthesia. The results are summarized in Figures 15-17.

[0156] The results showed that the effect of placing TE-8105 under the tongue for 1 minute in reducing blood glucose was substantially below the optimal level, and 3 minutes was not ideal for the absorption of TE-8105. The effect tended to stabilize after placing TE-8105 under the tongue for 5, 7, and 10 minutes (Figure 15). Weight loss was observed in db / db mice treated with TE-8105 under the tongue for 5, 7, and 10 minutes, but no weight loss was observed in the 1- or 3-minute treatment groups (Figure 16). Furthermore, when the liquid formulation of TE-8105 was administered sublingually to db / db mice for 5 or 7 minutes, it reduced the mice's food and water intake (Figure 17).

[0157] Example 8: Comparison of the effects of once-daily sublingual administration of different pH values ​​to TE-8105 mice (up to db / db) for five days on serum glucose, body weight, and food and water intake.

[0158] In this embodiment, the effect of the pH of the liquid formulation on the efficiency of sublingual mucosal absorption of TE-8105 was investigated. Accordingly, liquid formulations of TE-8105 with pH values ​​of 6.5, 7.0, 7.5, 8.0, and 8.5 were prepared and administered sublingually once daily to db / db mice for five days. The results are shown in Figures 18-21.

[0159] As shown in Figure 18, the TE-8105 liquid formulation at pH 7.0 was most effective in lowering blood glucose levels, followed by the pH 7.5 formulation, while formulations at pH 6.5, 8.0, and 8.5 were less effective. Furthermore, the TE-8105 liquid formulations at pH 7.0 and 7.5 were most effective for weight loss, while the formulations at pH 6.5, 8.0, and 8.5 were ineffective (Figure 19). In addition, TE-8105 at pH 7.0 was most effective in reducing food intake (Figure 20), and the TE-8105 liquid formulations at pH 7.0 and 7.5 showed statistically significant differences in reducing water intake compared to formulations at other pH levels (Figure 21).

[0160] Example 9: Effect of sublingual administration of TE-8105 and smegglutinin at 2 QD on the accumulation and decline of serum glucose in db / db mice.

[0161] As described in Examples 2 and 3, both TE-8105 and smegglutinin, administered sublingually, exhibited similar bioavailability of 7-8%. In this example, we investigated whether the relative potency or half-life of TE-8105 and smegglutinin affected their efficiency and kinetics in controlling serum glucose in db / db mice. To achieve this, db / db mice were administered TE-8105 or smegglutinin sublingually twice daily (2QD), and serum glucose levels were measured after 128 hours. The results are illustrated in Figure 22.

[0162] The results confirm that semaglutide can maintain blood glucose levels at normal levels, similar to the effect of frequent administration of TE-8105; however, during the gradual tapering period after discontinuation, the blood glucose levels in the semaglutide group returned to abnormal levels more quickly than in the TE-8105 group (Figure 22).

[0163] Example 10: Effect of different doses of TE-8105 or smegglutinin administered at 2QD on serum glucose levels in db / db mice

[0164] In this embodiment, the effects of twice-daily sublingual administration of different doses of TE-8105 and smegglutinin on serum glucose levels in db / db mice were investigated. The results are shown in Figure 23.

[0165] The results showed that a higher dose of semaglutide (350 nmol / kg) administered sublingually twice daily was required to lower blood glucose to the same level as TE-8105 (200 nmol / kg) administered twice daily (Figure 23). These observations are consistent with those of the two GLP-1 RA-Ps administered subcutaneously, as 50 nmol / kg semaglutide and 30 nmol / kg TE-8105 had similar efficacy in lowering blood glucose.

[0166] Example 11: Effects of prolonged once-daily (QD) or once-every-two-day (Q2D) sublingual administration of TE-8105 and smegglutinin on serum glucose control, body weight reduction, water and food intake in db / db mice

[0167] As described in Examples 2 and 3, one of the main objectives of this disclosure is to estimate the bioavailability between sublingual and subcutaneous delivery of GLP-1 RA-P. Since the first two sublingual doses on the first day are only a few hours apart (7 hours), drug clearance is not high. In this example, the feasibility and relative advantages of extended once-daily (QD) or once-every-two-day (Q2D) sublingual delivery of TE-8105 and semaglutide are explored, as semaglutide has been approved by regulatory agencies for the treatment of type 2 diabetes and obesity. An injection-free oral semaglutide formulation (trade name "Rebex") has been approved, setting a state-of-the-art benchmark in the field of GLP-1 RA-P drug delivery.

[0168] To simulate potential dosing regimens in human individuals, db / db mice were administered TE-8105 and semaglutide at a rate extended once daily (QD) or every two days (Q2D) for a total of 7 or 4 doses, respectively. Serum glucose levels were measured daily (Figure 24A) and the relative AUC of serum glucose levels over time was compared (Figure 24B). Body weight was measured between different groups (Figure 25), and food and water intake (Figure 26) was measured for 11 days, including the dose reduction period after the last administration. Results showed that extended once-daily (QD) administration of 200 nmol / kg of semaglutide and TE-8105 effectively controlled serum glucose levels, reduced body weight, and decreased water and food intake. However, every two days (Q2D) administration of TE-8105 still effectively controlled serum glucose levels, reduced body weight, and decreased water and food intake, while semaglutide did not exhibit these effects under the same administration conditions. Based on experiments with subcutaneous administration of GLP-1 RA-P, a smooth Q2D treatment profile in human individuals can be converted into a smooth QW treatment profile.

[0169] Example 12: Effects of TE-8105 administration for 10 days using the 2QD regimen on serum glucose, body weight, and food and water intake in ZFDM rats.

[0170] In this embodiment, the effects of TE-8105 on serum glucose levels, body weight, food and water intake in ZFDM rats were investigated. The results are shown in Figures 27 to 30.

[0171] The results showed that a single subcutaneous injection of TE-8105 at 15 nmol / kg reduced blood glucose by 25% within 2 days, and the blood glucose level gradually recovered after several days. Two administration regimens of TE-8105 are provided: (1) 6 and 14 doses of TE-8105 at 200 nmol / kg, and (2) 6 doses of TE-8105 at 200 nmol / kg and 14 doses of TE-8105 at 400 nmol / kg. Treatment was administered twice daily for 10 days, and rats were monitored for 8 days after the treatment.

[0172] The results showed that subcutaneous injection of TE-8105 caused a decrease in rat body weight within one day, followed by a sustained increase over the next 16 days. On the other hand, both sublingual administration regimens of TE-8105 caused a gradual decrease in rat body weight within 10 days of administration, followed by a recovery and increase in body weight 8 days after discontinuation of administration (Figure 27).

[0173] The changes in food and water intake after subcutaneous injection and two sublingual administration regimens almost mirrored the changes in body weight. AUC plots further showed that both sublingual administration regimens reduced blood glucose, body weight, and food and water intake, with better results than a single subcutaneous injection (Figures 28–30).

[0174] Example 13: Effect of sublingual administration of TE-8105 containing menthol and glycerol for 2 days according to the 2QD regimen on the in vivo glucose content of db / db mice.

[0175] In this embodiment, a liquid formulation of TE-8105, with or without additional peppermint oil and glycerin, was prepared according to the components listed in Table 1. The formulation prepared by the aforementioned method was then administered to db / db mice at a designed dose twice daily via sublingual or subcutaneous injection for two days, and serum glucose levels were measured over a 120-hour period. The results are illustrated in Figure 31.

[0176] Table 1. Liquid formulation of TE-8105 containing menthol and glycerin

[0177] The results showed that sublingual administration of the TE-8105 liquid formulation containing menthol and glycerin significantly reduced serum glucose levels compared to subcutaneous injection of TE-8105. However, whether the inclusion of menthol and glycerin truly has a statistically significant additional benefit on serum glucose levels requires further investigation.

[0178] Example 14: Effects of sublingual administration of TE-8105 soft gel on serum glucose levels, body weight, food and water intake in db / db mice

[0179] In this embodiment, the effects of TE-8105 in soft gel form on serum glucose, body weight, and food and water intake in db / db mice were investigated. To achieve this, TE-8105 in liquid form was mixed with 5% or 10% gelatin derived from cold-water fish or pig to form a disc-shaped TE-8105 soft gel. The TE-8105 soft gel prepared in this way was administered sublingually to db / db mice twice daily for 3 days, and serum glucose levels, body weight, and food and water intake were measured. The results are shown in Figures 32 to 34.

[0180] The results showed that when the soft gel was delivered into the sublingual space of db / db mice, the mice melted within 1-2 minutes. Furthermore, TE-8105 soft gel mixed with 5% or 10% gelatin significantly reduced blood glucose levels, with better results than the control group containing gelatin (without TE-8105). Data showed that TE-8105 containing 5% gelatin was more effective at reducing blood glucose levels than TE-8105 containing 10% gelatin (Figure 32). In addition, the pattern of weight loss reflected the pattern of glucose reduction (Figure 33); and the pattern of reduced food intake also reflected the pattern of reduced water intake (Figure 34).

[0181] Example 15: Effects of sublingual administration of a soft gel formed from a mixture of smegglutinin liquid formulation and gelatin on serum glucose levels, body weight, food and water intake in db / db mice.

[0182] In this embodiment, the effects of smegglutinin in the form of a soft gel on serum glucose levels, body weight, and food and water intake in db / db mice were investigated. The soft gel (i.e., a liquid formulation of smegglutinin and fish-derived gelatin) was prepared in the same manner as the TE-8105 soft gel described in Example 14. The soft gel was administered sublingually twice daily for 3 days, and serum glucose, body weight, and food and water intake were measured. The results are illustrated in Figures 35 to 37.

[0183] The results showed that the colloid composed of smegglutinin and 10% gelatin significantly lowered blood glucose levels compared to the control group (which used gelatin colloid without smegglutinin) (Figure 35). Furthermore, the pattern of weight loss reflected the pattern of glucose reduction (Figure 36); the pattern of reduced food intake also reflected the pattern of reduced water intake (Figure 37).

[0184] Example 16: Effects of sublingual administration of tablets made from TE-8105 liquid formulation and crospovidone on serum glucose levels, body weight, food and water intake in db / db mice

[0185] In this embodiment, TE-8105 tablets suitable for sublingual administration were prepared. To achieve this, a liquid formulation of TE-8105 was mixed with crospovidone (a highly cross-linked modified form of polyvinylpyrrolidone (PVP), commonly used as a disintegrant for tablets). The TE-8105 tablets prepared by the aforementioned method were administered sublingually to db / db mice twice daily for three days. However, since the db / db mice were anesthetized during tablet delivery and their tongues and mouths remained fixed, there was no saliva in the sublingual region to disintegrate the tablets. Accordingly, a small amount of distilled water was added to the sublingual region to assist in tablet disintegration. The results are illustrated in Figures 38-40.

[0186] The results showed that TE-8105 tablets containing 1% crospovidone could reduce high blood glucose levels in diabetic db / db mice to normal levels (Figure 38). Furthermore, TE-8105 tablets effectively reduced body weight and decreased food and water intake in obese db / db mice (Figures 39 and 40).

[0187] Example 17: Effects of sublingual administration of tablets made from smegglutinin liquid formulation and crospovidone on serum glucose levels, body weight, food and water intake in db / db mice

[0188] In this embodiment, semaglutide tablets (i.e., semaglutide liquid formulation mixed with 1% crospovidone) were prepared and administered according to a procedure similar to that described in Example 16, and the effect of the tablets on blood glucose control was monitored. The results are illustrated in Figures 41 to 43.

[0189] The results showed that semaglutide tablets effectively reduced blood glucose levels in db / db mice to normal levels (Figure 41). Furthermore, semaglutide tablets also effectively reduced body weight (Figure 42) and food and water intake (Figure 43).

[0190] Example 18: Effects of sublingual administration of TE-1805 menthol tablets on serum glucose levels, body weight, food and water intake in db / db mice

[0191] In this embodiment, TE-8105 peppermint tablets were prepared by adding 0.1%, 0.5%, or 1.0% peppermint oil to a TE-8105 liquid formulation containing 1% crospovidone, following the procedure described in Example 16. The effect of TE-8105 peppermint tablets on blood glucose control in db / db mice was monitored. The results are shown in Figures 44 to 47.

[0192] The results showed that TE-8105 tablets without peppermint oil and those containing 0.1%, 0.5%, or 1.0% peppermint oil all effectively reduced blood glucose levels, with the tablets containing 0.5% peppermint oil showing the best efficacy (Figure 44). Furthermore, TE-8105 tablets without peppermint oil and those containing 0.1%, 0.5%, or 1.0% peppermint oil all effectively reduced body weight, with the tablets containing 0.1% and 0.5% peppermint oil showing better efficacy (Figure 45). Regarding food intake, TE-8105 tablets containing 0.1% and 0.5% peppermint oil significantly reduced food intake in db / db mice; other groups also showed a trend towards reduced food intake (Figure 46). Regarding water intake, all TE-8105 tablets reduced water intake, with the tablets containing 0.1% and 0.5% peppermint oil showing the best efficacy (Figure 47).

[0193] Example 19: Bioavailability assessment of TE-8105, smegglutinin, and telpoxetine in human individuals based on a mouse model.

[0194] The results of Examples 2-18 confirm that when mice are maintained in anesthesia for approximately 5-7 minutes, the delivered liquid containing GLP-1 RA-P can be retained in the sublingual region of db / db mice. Currently, no publicly available procedures demonstrate that a liquid form of drug administered to the sublingual region can be retained intact in that region for an extended period. Literature review indicates that humans can retain small amounts of liquid in the sublingual cavity for a short period (5-10 minutes) without expelling the liquid due to oral or tongue movements. Furthermore, published literature indicates that 1.5 to 2.0 ml of liquid delivered to the sublingual cavity of a human individual is the maximum appropriate volume. In this disclosure, the optimal time for sublingual absorption of TE-8105 in db / db mice is confirmed to be 5 to 10 minutes (Example 7).

[0195] In the experiments described herein, a stock solution of TE-8105 and semaglutide at a concentration of 5 mg / mL was prepared from powder. The stock solution was then diluted to other concentrations using a liquid formulation buffer. TE-8105 drug product supplied by a contracted CDMO was also used in this study, with a liquid formulation concentration of 5 mg / mL. The results showed that the optimal concentration for promoting transmucosal transport in the sublingual region was 3–5 mg / mL (Example 6). The effect of concentration on transmucosal absorption was related to molecular interactions between GLP-1 RA-P and molecules on the surface of mucosal epithelial cells. These findings are consistent with the solubility of TE-8105 (5 mg / mL) and semaglutide (3.2 mg / mL) in aqueous buffer. Semaglutide exhibits very high solubility (30 mg / mL) in aqueous buffer due to its nine terminal amino acid residues. However, transmucosal transport may be limited to an optimal concentration of 3–5 mg / mL.

[0196] The solubility of TE-8105 is based on the drug product presented in liquid formulation (5 mg per mL) as demonstrated in Phase I and IIA clinical trials. The solubility of the semaglutide liquid formulation for subcutaneous injection is based on the highest known dose form (0.24 mg per 0.75 mL). Therefore, 2 mL of TE-8105 liquid formulation may contain 10 mg; 2 mL of semaglutide liquid formulation may contain 6.4 mg; and 2 mL of telpolide liquid formulation should contain 10 mg.

[0197] In Table 2, the required doses and frequencies for sublingual delivery of TE-8105, smegglutide, and telpoxetine are based on the expected range of maintenance doses for these drugs administered subcutaneously and in human individuals. Bold text indicates feasible conditions, assuming the expected bioavailability. For each GLP-1 RA-P with an expected bioavailability, only dosage conditions that do not require frequent dosing are listed. For example, if 10 mg once weekly is feasible, this condition is listed, while the options of 5 mg twice weekly or 1.4 mg daily are not listed; similarly, if 4.6 mg every two days is feasible, this condition is listed, while the options of 2.3 mg daily and 4.6 mg twice daily are not listed.

[0198] The frequency is labeled as follows: 2QD: Twice daily QD: Once a day Q2D: Once every two days 2QW: Twice a week QW: Once a week Q2W: Every two weeks Table 2. Dosage and frequency of sublingual administration of TE-8105, smegglutinin, and telpoxetine.

[0199] Note: Options in bold are feasible under the expected bioavailability conditions.

[0200] Expected bioavailability is the percentage of bioavailability achievable with subcutaneous delivery.

[0201] In summary, the data disclosed herein confirm that the sublingual formulation of this disclosure, when administered to an individual in need, unexpectedly achieves a high bioavailability of approximately 7-10% for GLP-1 RA-P, or 2-10% based on potential individual variability, where this bioavailability is relative to that achieved by subcutaneous injection. Furthermore, it reduces an individual's blood glucose levels, body weight, and food and water intake, thus making it suitable for the treatment of obesity, type II diabetes, or related conditions.

[0202] To make the description of this disclosure more detailed and complete, illustrative descriptions of embodiments and specific examples of the present invention are provided below; however, these are not the only forms of implementing or utilizing the specific examples of the present invention. The embodiments cover features of multiple specific examples and methods and steps for constructing and operating these specific examples, as well as their order. However, other specific examples may also be used to achieve the same or equivalent functions and order of steps.

Claims

1. A pharmaceutical formulation for treating obesity, type II diabetes, or related diseases, comprising: 5-10 mg of glucagon-like peptide-1 receptor agonist peptide (GLP-1 RA-P) was dissolved in 1.5-2.0 mL of buffer solution; in The GLP-1 RA-P is selected from the group consisting of TE-8105, smegglutinin and telposide, wherein the structure of TE-8105 is shown in Figure 1. The pH of this pharmaceutical formulation is between 5.5 and 7.5, and it is suitable for sublingual administration; and Compared to administration to an individual via subcutaneous injection, this pharmaceutical formulation achieves a bioavailability of 2-10% for GLP-1 RA-P.

2. The pharmaceutical formulation of claim 1, wherein the pharmaceutical formulation is in liquid, tablet or gel form.

3. The pharmaceutical formulation of claim 2, wherein the pharmaceutical formulation is in liquid form and contains 5-10 mg of TE-8105 dissolved in 1.5-2.0 mL of buffer solution.

4. The pharmaceutical formulation of claim 2, wherein the pharmaceutical formulation is in liquid form and contains 5-6 mg of smegglutide dissolved in 1.5-2.0 mL of buffer solution.

5. The pharmaceutical formulation of claim 2, wherein the pharmaceutical formulation is in liquid form and contains 5-10 mg of telpoide dissolved in 1.5-2.0 mL of buffer solution.

6. The pharmaceutical formulation of claim 2 further comprises 0.1% (w / v) peppermint oil.

7. The pharmaceutical formulation as described in claim 2 further comprises 5-10% gelatin, and the pharmaceutical formulation is in the form of a soft gel.

8. The pharmaceutical formulation of claim 2 further comprises 1-10% crospovidone, and the pharmaceutical formulation is in tablet form.

9. The pharmaceutical formulation of claim 1, wherein the disease associated with obesity and type II diabetes is overweight, fatty liver, non-alcoholic fatty liver disease, diabetic cardiomyopathy, or coronary atherosclerotic heart disease.

10. A method for treating an individual's obesity, type II diabetes, or related diseases, comprising: One effective dose of the pharmaceutical formulation, containing GLP-1 RA-P, is administered sublingually to the individual and maintained sublingually for 5-10 minutes. The GLP-1 RA-P is selected from the group consisting of TE-8105, smegglutinin and telposide, wherein TE-8105 has the structure shown in Figure 1. The pH value of this pharmaceutical formulation is between 5.5 and 7.5; The pharmaceutical formulation is administered to the individual at a frequency of once daily, twice daily, once every two days, once weekly, or twice weekly; and Compared to administration to an individual via subcutaneous injection, this pharmaceutical formulation achieves a bioavailability of 2-10% for GLP-1 RA-P.

11. The method of claim 10, wherein the pharmaceutical formulation is in the form of a liquid, tablet, or gel.

12. The method of claim 11, wherein This pharmaceutical formulation is in liquid form and contains 5-10 mg of TE-8105 dissolved in 1.5-2.0 mL of buffer solution; and The pharmaceutical formulation is administered to the individual at a frequency of twice daily, once daily, once every two days, twice weekly, or once weekly.

13. The method of claim 11, wherein This pharmaceutical formulation is in liquid form and contains 5-6 mg of smegglutide dissolved in 1.5-2.0 mL of buffer solution; and The pharmaceutical formulation is administered to the individual at a frequency of twice daily, once daily, or once every two days.

14. The method of claim 11, wherein This pharmaceutical formulation is in liquid form and contains 5-10 mg of telpolide dissolved in 1.5-2.0 mL of buffer solution; and The pharmaceutical formula is administered to the individual twice daily.

15. The method of claim 11, wherein the pharmaceutical formulation further comprises 0.1-0.5% (w / v) peppermint oil.

16. The method of claim 11, wherein the pharmaceutical formulation further comprises 5-10% (w / v) gelatin in a soft gel form.

17. The method of claim 11, wherein the pharmaceutical formulation further comprises 1-10% crospovidone and is in tablet form.

18. The method of claim 10, wherein the disease associated with obesity and type II diabetes is overweight, fatty liver, non-alcoholic fatty liver disease, diabetic cardiomyopathy, or coronary atherosclerotic heart disease.

19. The method of claim 10, wherein the individual is a human being.

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