Pharmaceutical composition of semaglutide and salt thereof for intranasal administration

Abstract

Pharmaceutical compositions of semaglutide or salts thereof for intranasal administration for the treatment of diabetes, obesity, non-alcoholic fatty liver disease (NAFLD) or neurodegenerative diseases. The method of delivering semaglutide through the intranasal route avoids repeated injections and improves systemic absorption of semaglutide as compared to oral pastilles.

Description

Technical Field

[0001] The present invention relates to pharmaceutical compositions of semaglutide and its salts for intranasal administration, and their use in the treatment of diabetes, obesity, non-alcoholic fatty liver disease (NAFLD) or neurodegenerative diseases. Background Art

[0002] Semaglutide (C 187 H 291 N 45 O 59 ) is a long-acting GLP-1 receptor agonist with 94% structural similarity to native GLP-1 (Knudsen et al., 2019), and it is also known as N6,26-{18-[N-(17-carboxyheptadecanoyl)-L-glutamyl]-10-oxo-3,6,12,15-tetraoxa-9,18-diazaoctadecanoyl}-[8-(2-amino-2-propanoic acid),34-L-arginine] human glucagon-like peptide 1 (7-37), as described in WO2020 / 084126. The preparation method of semaglutide is described in Example 4 of WO2006 / 097537. Semaglutide can exist in the composition in a fully or partially ionized form.

[0003] Semaglutide is an anti-diabetic drug used for the treatment of type 2 diabetes and long-term weight management. The action of semaglutide is similar to that of human glucagon-like peptide-1 (GLP-1), which increases insulin secretion, thereby increasing glucose metabolism. It has been found that semaglutide can reduce hyperglycemia, body weight, steatosis, and improve the cognitive ability of patients with neurodegenerative diseases (Mahapatra et al, 2022). Currently, two semaglutide products have been approved by the US Food and Drug Administration (USFDA), Health Canada, the European Medicines Agency, and the Japanese Health Ministry for the treatment of type 2 diabetes, including (subcutaneous injection, once a week; available in 0.5 and 1.0 mg specifications) and (oral tablets, once a day; available in 3, 7, and 14 mg specifications).

[0004] The use of the injection pen is relatively complex and requires experienced medical staff to train patients. Long-term and frequent injection of semaglutide will bring inconvenience and discomfort to patients and pose a risk of infection and adverse reactions (such as erythema) to the injection site (please refer to FDA Package Insert). Semaglutide oral tablets provide an alternative treatment option for patients who are unwilling or unable to self-inject antidiabetic drugs; however, despite the above advantages, due to the low permeability of the semaglutide molecule and extensive degradation and metabolism in the gastrointestinal (GI) tract, the oral bioavailability of semaglutide is very low (0.4 - 1%), and there is a large individual difference. To achieve a comparable systemic absorption to injected semaglutide, the weekly dose of oral semaglutide tablets is 49 to 98 mg, which is much higher than the weekly dose of 0.5 to 1.0 mg for injection (about 100 times); the oral tablets must be taken daily at least 30 minutes before the first meal or drink; at the same time, each tablet contains up to 300 mg of the permeation enhancer (SNAC), which may cause gastrointestinal adverse reactions such as nausea, abdominal pain, and vomiting (please refer to FDA Package Insert).

[0005] Given the disadvantages of commercially available semaglutide products, nasal administration is an alternative and promising option because the drug can directly enter the blood circulation from the absorption site, thus completely avoiding degradation in the gastrointestinal tract and hepatic metabolism, while being convenient for patients to use and more acceptable to the public. The advantages of intranasal semaglutide administration include but are not limited to: non-invasive, easy to use, and patients can self-administer; the drug directly enters the blood circulation through the nasal epithelial mucosa, is rapidly and completely absorbed, and has a rapid onset of action; the bioavailability is increased due to avoiding GI degradation and hepatic metabolism; compared with oral tablets, the intranasal dosage is small and the dosing frequency is reduced (i.e., 1 - 2 doses per week); local or systemic adverse reactions are effectively controlled; at the same time, there are no drug interactions and food effects; no dose adjustment is required for special populations.

[0006] WO2007 / 146488 discloses a nasal liquid composition of the GLP-1 agonist liraglutide. To maintain the physicochemical stability of the preparation, the pH is adjusted to 8.5 or higher, which is much higher than the pH of nasal mucus under normal physiological conditions (5.5 - 6.5), and long-term use will cause various local adverse reactions. In addition, liraglutide is a short-acting GLP-1 agonist, and the subcutaneous injection dose is 1.25 - 1.9 mg per day. According to the pharmacokinetic results of the drug in rabbits in the patent specification, the inventors concluded that to achieve the same absorption as injection, the intranasal preparation must reach a drug concentration of 50 mg / mL and be administered 2 - 4 doses per day. Frequent nasal administration may cause toxic effects and poor patient compliance.

[0007] WO2007 / 0611434 describes a pharmaceutical composition of exenatide for intranasal administration, which uses a formulation technique similar to that in WO2007 / 146488, plus a dipeptidyl peptidase (DPP) IV inhibitor to minimize the enzymatic degradation of exenatide. Due to the short half-life of exenatide (2.4 hours), the effective drug concentration in the blood circulation and the hypoglycemic effect can only be maintained for 3-4 hours. To maintain the therapeutic effect, at least 6-8 times of intranasal administration are required every day. Therefore, this composition is not suitable for clinical use in the treatment of metabolic syndrome in mammals. Summary of the Invention

[0008] The present invention relates to a pharmaceutical composition of semaglutide and its salts for intranasal administration, which is used to treat diabetes, obesity, non-alcoholic fatty liver disease (NAFLD) or neurodegenerative diseases by intranasal administration. Compared with commercially available injection pens or oral tablets, delivering semaglutide through the intranasal route avoids long-term injections and improves systemic absorption. Brief Description of the Drawings

[0009] Figure 1 . Mean plasma concentration-time curve of semaglutide in rats after intranasal administration of PT-N01, PT-N02, and PT-N03 at a single dose of 0.2 mg / animal.

[0010] Figure 2 . Mean plasma concentration-time curve of semaglutide in rabbits after intranasal administration of PT-N01, PT-N04, and PT-N05 at a single dose of 4 mg / animal and subcutaneous injection of PT-S01 at a single dose of 0.25 mg / animal.

[0011] Figure 3 . Mean blood glucose-time curve of rabbits after intranasal administration of PT-N01, PT-N04, and PT-N05 at a single dose of 4 mg / animal and subcutaneous injection of PT-S01 at a single dose of 0.25 mg / animal.

[0012] Figure 4 . Mean body weight-time curve of rabbits after intranasal administration of PT-N01, PT-N04, and PT-N05 at a single dose of 4 mg / animal and subcutaneous injection of PT-S01 at a single dose of 0.25 mg / animal.

[0013] Figure 5 . Mean food intake-time curve of rabbits after intranasal administration of PT-N01, PT-N04, and PT-N05 at a single dose of 4 mg / animal and subcutaneous injection of PT-S01 at a single dose of 0.25 mg / animal.

[0014] Figure 6. Mean plasma concentration-time curve of semaglutide after intranasal administration of PT-N06 and oral administration of tablets at a single dose of 7 mg / animal. Detailed implementation mode

[0015] The present invention relates to a pharmaceutical composition capable of delivering a sufficient dose of semaglutide through the intranasal route. The marketed oral semaglutide causes some adverse reactions, such as high first-pass effect and extremely low bioavailability, high inter-subject variability, drug-drug interactions and food effects, and gastrointestinal adverse reactions caused by penetration enhancers. The intranasal composition of the present invention enables rapid absorption of semaglutide through the nasal mucosa, and completely avoids gastrointestinal degradation and metabolism, thus well solving the above-mentioned disadvantages of oral administration. Intranasal drug absorption is rapid and complete, convenient to use, and has few side effects.

[0016] The composition according to the present invention comprises an active ingredient, namely semaglutide or a pharmaceutically acceptable salt thereof. The semaglutide used in the present invention includes the free base form or a pharmaceutically acceptable salt form. Pharmaceutically acceptable salts include, but are not limited to, sodium, potassium, calcium, magnesium, lithium, cesium, palladium, ammonium. The salt preferably used in the present invention is semaglutide sodium, which is formed by semaglutide and sodium in a mass ratio of 1:1 to 100:1.

[0017] The use of semaglutide or a pharmaceutically acceptable salt thereof according to the present invention includes a preparation in which a therapeutic dose of semaglutide is delivered to the nasal mucosa. The preferred dosage form is a liquid dosage form, including solutions, suspensions, emulsions, bioadhesives or in-situ gels, microspheres, nanoparticles, self-emulsifying drug delivery systems; or solid dosage forms, including powders, granules; or semi-solid dosage forms, including ointments, creams, hydrogels; or other forms suitable for intranasal delivery in the art.

[0018] Semaglutide is a hydrophilic compound with a relatively high molecular weight (MW: 4113.64), exceeding the upper cut-off molecular weight for intranasal mucosal delivery (Pathak K., 2011), so the nasal absorption of semaglutide is very low. In our pharmacokinetic and pharmacodynamic studies in rabbits, the intranasal bioavailability of semaglutide aqueous solution was only 0.09%, and even when a high dose of 4 mg was administered intranasally to each animal, the drug blood concentration was far below the minimum effective concentration; therefore, it is imperative to use various mucosal drug absorption promotion techniques to improve intranasal absorption.

[0019] Surprisingly, the present invention shows that the bioavailability of semaglutide can be effectively improved by comprehensively using a variety of absorption promoters. The absorption promoters include but are not limited to: i) cell-penetrating peptides (CPPs) as carriers to achieve transcellular drug transport; ii) tight junction modulators that increase paracellular penetration; iii) bioadhesive polymers that prolong the residence time in the nasal mucosa and minimize the clearance of drugs by nasal cilia.

[0020] An important aspect of the present invention is to promote the transcellular transport of semaglutide by means of cell-penetrating peptides (CPPs). It has been found that the nasal absorption of semaglutide can be significantly improved by physically mixing or covalently coupling the semaglutide molecule with cell-penetrating peptides. Cell-penetrating peptides are positively charged short peptides with a length of 5-30 amino acids, which can penetrate biological membranes and be used as a new type of carrier for intracellular delivery (Derakhshankhah H, Jafari S. 2018). Cell-penetrating peptides have been widely studied due to their high intracellular transport efficiency and low cytotoxicity and can be used to deliver macromolecules. Preferred cell-penetrating peptides of the present invention include but are not limited to TAT, R6, R8, penetratin, protamine, transportin and their derivatives. The cell-penetrating peptides can form complexes with semaglutide and significantly enhance the delivery across the nasal mucosal epithelial cells through the mechanism of endocytosis.

[0021] Another important aspect of the present invention is to further improve the paracellular transport of semaglutide across the nasal mucosa by tight junction modulators. The tight junction modulators of the present invention can be selected from: (i) phospholipid surfactants, including dodecylphosphocholine (DPC), 1,2-didecylphosphatidylcholine (DDPC), 1,2-distearoyl-sn-glycero-3-phosphocholine (DSPC), 1-didecanoyl 1-sn-glycero-3-phosphocholine (LLPC), 1,2-dioctanoyl-sn-glycero-3-phosphocholine (D8PC), 1-1-palmitoyl-2-glutaroyl-sn-glycero-3-phosphocholine (PGPC), and the preferred phospholipid surfactants are DPC and DSPC because of their better solubility and stability in liquid formulations and low mucosal toxicity; (ii) cyclodextrin derivatives, including α-cyclodextrin, β-cyclodextrin, dimethyl-β-cyclodextrin, hydroxypropyl-β-cyclodextrin, γ-cyclodextrin, and the preferred cyclodextrin derivatives are dimethyl-β-cyclodextrin and hydroxypropyl-β-cyclodextrin; (iii) chelating agents, including EDTA, EGTA, BAPTA, and preferably EDTA; (iv) bile salts, including sodium cholate, dehydrocholate, taurocholate; (v) fatty acids and phosphate esters, including oleic acid, sodium decanoate, palmitoyl carnitine, lysophosphatidic acid; (vi) cationic polymers, such as chitosan derivatives and protamine; (vii) surfactants, including sodium dodecyl sulfate (SDS), Tween 20; (viii) nitric oxide donors or bradykinin.

[0022] A major obstacle to nasal delivery is the rapid clearance of drug formulations (aqueous solutions or dry powders) from the nasal cavity by the rapidly moving nasal cilia, with a clearance half-life of approximately 15 minutes, resulting in low bioavailability (Merkus et al., 1998). Especially for semaglutide and other polypeptide drugs with high molecular weight and low nasal mucosa permeability, nasal ciliary clearance can lead to extremely low bioavailability. Therefore, components with mucoadhesive properties, including bioadhesives / mucoadhesives and / or in-situ gelling agents, should be considered to increase the residence time of the drug in the nasal cavity and prolong the drug release time to achieve a sustained therapeutic effect.

[0023] In some embodiments of the present invention, pharmaceutically acceptable bioadhesives / mucoadhesives are selected from methylcellulose, hydroxymethylcellulose, hydroxypropylcellulose, hydroxypropylmethylcellulose, carboxymethylcellulose, hyaluronic acid, sodium alginate, chitosan, gelatin, lectin, poly(acrylic acid), gum arabic, carbomer 934P, xanthan gum, guar gum, and carrageenan, and combinations thereof.

[0024] In-situ gelling agents are water-soluble polymers with mucoadhesive properties that can change their rheological behavior in response to relevant changes such as ions, pH, and temperature. The liquid drug forms a free-flowing non-Newtonian fluid after mixing in the spray device and forms a thick gel when mixed or atomized. In some embodiments of the present invention, pharmaceutically acceptable in-situ gelling agents are selected from poloxamer, gellan gum, pectin, carbomer, carrageenan, cellulose acetate phthalate, and combinations thereof.

[0025] The first aspect relates to a pharmaceutical composition comprising semaglutide in a dose of 0.01 mg to 100 mg for a suitable formulation. The composition is suitable for intranasal administration, typically by means of a nasal spray device.

[0026] Pharmaceutically acceptable buffers can be used to maintain optimal pH conditions to obtain physicochemical stability and minimize local irritation to the nasal mucosa. According to the present invention, a suitable pH range is from 3.0 to 9.0, preferably from 4.0 to 7.0. Preferred buffer systems include, but are not limited to, phosphate buffer, acetate buffer, borate buffer, citrate buffer, tartrate buffer, and tris buffer.

[0027] The composition also contains one of the drug preservatives, including but not limited to: benzalkonium chloride, benzethonium chloride, benzyl alcohol, chlorobutanol, chlorhexidine, methylparaben and propylparaben, phenethyl alcohol, phenylmercuric acetate, and thimerosal. Preservatives with no adverse reactions to the nasal mucosa are preferred, including but not limited to benzyl alcohol, benzalkonium chloride, chlorhexidine, and thimerosal.

[0028] Finally, the composition of the present invention may further comprise: (1) a chelating agent, namely sodium EDTA; (2) an antioxidant, namely sodium metabisulfite; (3) an osmotic pressure regulator, including glucose, glycerol, hydroxypropyl beta-cyclodextrin, mannitol, sorbitol, potassium chloride, and sodium chloride.

[0029] The semaglutide or its pharmaceutical salt composition, preferably in an aqueous or semi-solid form, is sprayed into the nasal cavity using an atmospheric pressure nasal spray device. Suitable spray devices include spray pumps and bottles, and can deliver a single dose or multiple doses of the drug by mechanical force. The spray volume for each nostril ranges from 10 to 200 μL, more preferably from 50 to 150 μL, and most preferably from 80 to 120 μL.

[0030] Another aspect of the present invention relates to the preparation of a therapeutic agent for the treatment of diabetes, obesity, non-alcoholic fatty liver disease (NAFLD), or neurodegenerative diseases, which comprises 0.1 to 50 mg of a therapeutic dose of semaglutide in a suitable pharmaceutical carrier for intranasal delivery.

[0031] The pharmacokinetic and pharmacodynamic profiles of semaglutide pharmaceutical compositions (PT-N01 to PT-N06) after intranasal administration to rats, rabbits, and beagle dogs are as described in Examples 3 to 5. To the inventors' knowledge, there are currently no patents or publications showing the delivery of semaglutide via the intranasal route. Compared with commercially available oral tablets The intranasal administration of semaglutide pharmaceutical compositions has unexpectedly rapid and significantly improved absorption. According to the animal pharmacokinetic results, the intranasal dose can be further reduced from 7 - 14 mg of oral semaglutide per day to 8 - 10 mg or less of intranasal semaglutide once a week, but the systemic absorption and efficacy will be comparable to or better than that of oral tablets, and it can control blood glucose and body weight faster, and there are no gastrointestinal toxicities, food effects, and drug interactions. Examples Example 1. Semaglutide Nasal Spray

[0032] PT-N01: Stir at ambient temperature to completely dissolve 200 mg of semaglutide in 10 mL of PBS 7.0 buffer solution. Then filter the solution through a 0.45 μm filter membrane and fill it into a glass bottle equipped with a metered spray pump for intranasal administration. Each spray will deliver 2 mg of semaglutide intranasally at a volume of 0.10 mL per spray.

[0033] PT-N02: In an exemplary composition of the present invention having the ingredients listed in Table 1, a cell-penetrating peptide - penetratin is used as a permeation enhancer to prepare a nasal spray composition. Table 1. Ingredients of PT-N02 Component Weight Unit Semaglutide 60 mg Permeatin 60 mg PBS 5.0 Buffer Q.s. to 6 mL mL

[0034] Preparation process: (a) Load 60 mg of semaglutide into a glass bottle equipped with a magnetic stir bar. (b) Add an appropriate amount of PBS 5.0 buffer and stir to dissolve the API at room temperature. (c) Add 60 mg of penetratin to the solution and stir the mixture until completely dissolved. (d) Check the pH and adjust the pH of the solution to 5.0 with HCl or NaOH solution. (e) Dilute to the required volume (5 mL) with PBS 5.0 buffer solution. (f) Filter the solution through a 0.45-micron filter. (g) Fill the solution into a 3.5-mL nasal spray bottle, and each press of the spray pump can deliver 1.2 mg of semaglutide (0.1 mL).

[0035] PT-N03: In an exemplary composition of the present invention, a nasal spray composition having the ingredients listed in Table 2 is prepared using penetration enhancers such as dodecylphosphocholine (DPC), 2,6-dimethyl-β-cyclodextrin, and disodium edetate (EDTA-2Na). Table 2. Ingredients of PT-N03 Component Weight Unit Semaglutide 100 mg Dodecylphosphocholine 300 mg 2,6-Dimethyl-β-cyclodextrin 1000 mg Disodium Edetate 100 mg Hydroxypropyl Methylcellulose 50 mg PBS 6.5 Buffer Q.s. to 10 mL mL

[0036] Preparation process: (a) Weigh semaglutide and load it into a glass bottle equipped with a magnetic stir bar. (b) Add an appropriate amount of PBS 6.5 buffer and stir until the API dissolves at room temperature. (c) Add dodecylphosphocholine to the solution and stir the mixture until completely dissolved. (d) Add 2,6-dimethyl-β-cyclodextrin and disodium edetate to the solution and stir until dissolved. (e) Add HPMC to the solution and dissolve the excipient by stirring. (f) Dilute to the required volume (10 mL) with PBS6.5 buffer solution. (g) Check the pH value and adjust the pH of the solution to 6.5 with HCl or NaOH solution. (h) Filter the solution through a 0.45-micron filter. (i) Fill the solution into a 3.5-mL nasal spray bottle, and each press of the spray pump can deliver 1 mg of semaglutide (0.1 mL).

[0037] PT-N04: In an exemplary composition of the present invention, a nasal spray composition having the ingredients listed in Table 3 is prepared using penetration enhancers such as dodecylphosphocholine (DPC), 2,6-dimethyl-β-cyclodextrin, and disodium edetate (EDTA-2Na). A bioadhesive polymer such as hydroxypropyl methylcellulose (HPMC K100LV) is applied to prolong the residence time of the drug in the nasal cavity. First, 200 mg of semaglutide is completely dissolved in about 8 mL of PBS 7.0 buffer solution at ambient temperature; then dodecylphosphocholine, 2,6-dimethyl-β-cyclodextrin, and disodium edetate are added and dissolved by stirring at ambient temperature; thereafter, HPMC is added and stirred until dissolved; finally, PBS 7.0 buffer solution is added to make up to 10 mL. The concentration of semaglutide in the final solution is 20 mg / mL. Table 3. Ingredients of PT-N04

[0038] After preparation, the drug solution is filtered through a 0.22 μm filter membrane and then filled into a metered nasal sprayer (0.10 mL per spray), and 2 mg of semaglutide is delivered by intranasal administration each time.

[0039] PT-N05: At ambient temperature, semaglutide and the absorption enhancer sodium N-(8-[2-hydroxybenzamido]) octanoate (SNAC) are completely dissolved in about 8 mL of PBS 7.0 buffer solution; then dimethyl-β-cyclodextrin and disodium edetate (EDTA-2Na) are added to the solution; hydroxypropyl methylcellulose (HPMC K100LV) is added to the solution and stirred at ambient temperature until completely dissolved; finally, an appropriate amount of PBS 7.0 buffer solution is added to the final solution containing the ingredients listed in Table 4 to make up to 10 mL. The concentration of semaglutide in the final spray solution is 20 mg / mL. Table 4. Ingredients of PT-N05

[0040] After preparation, the drug solution is filtered through a 0.22 μm filter membrane and then filled into a metered nasal sprayer (0.10 mL per spray), and 2 mg of semaglutide is delivered by intranasal administration each time.

[0041] PT-N06: Dissolve semaglutide and the absorption promoter dodecylphosphocholine (DPC) completely in approximately 8 mL of PBS 7.0 buffer solution at ambient temperature; then add disodium edetate (EDTA-2Na) to the solution; next, add hypromellose (HPMC K100LV) to the solution and stir at ambient temperature until completely dissolved; finally, add PBS 7.0 buffer solution to the final solution containing the components listed in Table 5 to 10 mL. The concentration of semaglutide in the final spray solution is 35 mg / mL. Table 5. Components of PT-N06

[0042] After preparation, filter the drug solution through a 0.22 μm filter membrane, and then fill it into a metered nasal sprayer (0.10 mL per spray), and deliver 3.5 mg of semaglutide intranasally each time. Example 2. Other Semaglutide Nasal Sprays

[0043] PT-B01: In a composition of the present invention having the components listed in Table 6, a bioadhesive semaglutide nasal spray is prepared using sodium carboxymethylcellulose (CMC-Na) as a thickening agent. Table 6. Components of PT-B01 Component Weight (per 100 mL) Semaglutide 2g Dodecylphosphocholine 3g Disodium Edetate 1g CMC-Na 0.5g PBS6.5 Buffer Q.s. to 100 mL

[0044] Preparation process: (a) Weigh semaglutide and place it in a glass bottle equipped with magnetic stirring. (b) Add phosphate buffer solution (pH 6.5) to the glass bottle and stir to dissolve the API at room temperature. (c) Add dodecylphosphocholine to the solution and stir the mixture at room temperature until dissolved. (d) Add disodium edetate and CMC-Na and stir to dissolve the excipients at room temperature. (e) Add PBS 6.5 buffer solution to the required volume (100 mL). (f) Check the pH and adjust the pH value of the solution to 6.0 - 7.0 with HCl or NaOH solution. (g) Filter the solution through a 0.45 μm filter. (h) Fill the solution into a 3.5 mL nasal sprayer, and deliver 2 mg of semaglutide intranasally each time (0.1 mL per spray).

[0045] PT-B02: In a composition of the present invention having the components listed in Table 7, a bioadhesive nasal spray is prepared using sodium hyaluronate. Table 7. Components of PT-B02 Component Weight (per 100 mL) Semaglutide 1g Dodecylphosphocholine 3g Disodium Edetate 1g Sodium Hyaluronate 0.2g PBS6.5 Buffer Q.s. to 100 mL

[0046] Preparation process: (a) Weigh dodecylphosphocholine (DPC) and place it in a glass bottle equipped with magnetic stirring. (b) Add an appropriate amount of PBS 6.5 buffer solution and stir the mixture at room temperature until dissolved. (c) Add semaglutide to the solution and stir at room temperature until completely dissolved. (d) Add disodium edetate to the solution and stir at room temperature to dissolve the excipient. (e) Add sodium hyaluronate to the solution and stir the mixed solution at room temperature until completely dissolved. (f) Add PBS 6.5 buffer solution to the required volume (100 mL). (g) Check the pH and adjust the pH of the solution to 6.0 - 7.0 with HCl or NaOH solution. (h) Filter the solution through a 0.45 - micron filter. (i) Load the solution tank into a 3.5 - mL nasal sprayer and administer 1 mg of semaglutide intranasally each time (0.1 mL per spray). Semaglutide nasal spray containing poloxamer:

[0047] PT - B03: In a composition of the present invention having the components listed in Table 8, a nasal spray composition is prepared using poloxamer. Table 8. Components of PT - B03 Component Weight (per 100 mL) Semaglutide 2g Dodecylphosphocholine 3g Disodium Edetate 1g Poloxamer 188 1g Poloxamer 407 18g PBS6.5 Buffer Q.s. to 100 mL

[0048] Preparation process: (a) Weigh semaglutide and place it in a glass bottle equipped with magnetic stirring. (b) Add an appropriate amount of PBS 6.5 buffer solution to the glass bottle and stir at room temperature to dissolve the API. (c) Add dodecylphosphocholine to the solution and stir the mixture at room temperature until completely dissolved. (d) Add disodium edetate to the solution and stir at room temperature to dissolve. (e) Add poloxamer 188 to the solution and stir in an ice bath until completely dissolved, then add Poloxamer 407 to the solution and stir in an ice bath to dissolve. (f) Add PBS 6.5 buffer solution to the required volume (100 mL). (g) Check the pH and adjust the pH of the solution to 6.0 - 7.0 with HCl or NaOH solution. (h) Place the solution at 4 °C overnight until a clear solution is obtained. (i) Filter the solution through a 0.45 μm filter. (j) Load the solution vial into a 3.5 mL nasal sprayer and administer 2 mg of semaglutide per nasal delivery (0.1 mL per spray). Semaglutide nasal spray containing Carbomer 934P.

[0049] PT-B04: In a composition of the present invention having the ingredients listed in Table 9, a nasal spray composition is prepared using Carbomer 934P. Table 9. Ingredients of PT-B04 Component Weight (per 100 mL) Semaglutide 2g Dodecylphosphocholine 3g Disodium Edetate 1g Carbomer 934P 0.2g PBS6.5 Buffer Q.s. to 100 mL

[0050] Preparation process: (a) Weigh semaglutide and place it in a glass bottle equipped with a magnetic stir bar. (b) Add an appropriate amount of PBS 6.5 buffer solution to the glass bottle and stir to dissolve the API at room temperature. (c) Add dodecylphosphocholine to the solution and stir the mixture until completely dissolved. (d) Add disodium edetate to the solution and stir the mixture until dissolved. (e) Add Carbomer 934P to the above solution and stir the mixture until completely dissolved. (f) Add PBS 6.5 buffer solution to the required volume (100 mL). (g) Check the pH and adjust the pH of the solution to 6.0 - 7.0 with HCl or NaOH solution. (h) Filter the solution through a 0.45 μm filter. (i) Load the solution vial into a 3.5 mL nasal sprayer and administer 2 mg of semaglutide per nasal delivery (0.1 mL per spray). Semaglutide nasal spray containing chitosan hydrochloride.

[0051] PT-B05: In a composition of the present invention having the ingredients listed in Table 10, a nasal spray composition is prepared using chitosan hydrochloride. Table 10. Ingredients of PT-B05 Component Weight (per 100 mL) Semaglutide 3.5g Dodecylphosphocholine 3g Disodium Edetate 1g Chitosan Hydrochloride 0.3g PBS6.5 Buffer Q.s. to 100 mL

[0052] Preparation process: (a) Weigh semaglutide and place it in a glass bottle equipped with a magnetic stir bar. (b) Add an appropriate amount of PBS 6.5 buffer solution to the glass bottle and dissolve the API by mixing at room temperature. (c) Add choline dodecyl phosphate and disodium edetate to the solution, and stir the mixture until completely dissolved. (d) Add chitosan·HCl to the solution, and stir to dissolve the excipient at room temperature. (e) Add PBS 6.5 buffer solution to the required volume (100 mL). (f) Check the pH, and adjust the pH of the solution to 6.0 - 7.0 with HCl or NaOH solution. (g) Pass the solution through a 0.45 - micron filter. (h) Load the solution tank into a 3.5 - mL nasal sprayer, and administer 3.5 mg of semaglutide intranasally each time (0.1 mL per spray). Semaglutide nasal spray containing hydroxypropyl cellulose.

[0053] PT - B06: In a composition of the present invention having the components listed in Table 11, a nasal spray composition is prepared using hydroxypropyl cellulose (HPC). Table 11. Components of PT - B06

[0054] Preparation process: (a) Weigh semaglutide and choline dodecyl phosphate, and place them in a glass bottle equipped with a magnetic stir bar. (b) Add an appropriate amount of PBS 6.5 buffer solution, and stir the mixture until completely dissolved at room temperature. (c) Add disodium edetate to the solution, and stir to dissolve. (d) Add hydroxypropyl cellulose to the solution, and stir to dissolve the excipient at room temperature. (e) Add phosphate buffer solution (pH 6.5) to the required volume (100 mL). (f) Check the pH, and adjust the pH of the solution to 6.0 - 7.0 with HCl or NaOH solution. (g) Pass the solution through a 0.45 - micron filter. (h) Load the solution tank into a 3.5 - mL nasal sprayer, and administer 3.5 mg of semaglutide intranasally each time (0.1 mL per spray). Semaglutide subcutaneous injection.

[0055] PT-S01: At ambient temperature, 20 mg of semaglutide was completely dissolved in 20 mL of PBS 7.0 buffer solution. Then the solution was filtered through a 0.22 μm filter membrane. Finally, the filtrate was filled into a glass bottle for subcutaneous injection. The concentration of the semaglutide injection was 1 mg / mL. Example 3. Pharmacokinetic study of semaglutide composition in rats.

[0056] This example aimed to study the nasal absorption of the semaglutide liquid preparation. SD rats were divided into 3 groups (3 rats per group), and each group was administered intranasally (0.2 mg / animal) with a solution composition (PT-N01, PT-N02, or PT-N03), which was prepared according to Example 1. During the experiment, multiple blood samples were collected from the rat tail vein until 24 hours. The concentration of semaglutide in rat plasma was determined by a validated LC-MS method. Pharmacokinetic parameters were generated by standard non-compartmental methods. Figure 1 Show the curves of the mean plasma concentration of semaglutide versus time after intranasal administration of PT-N01, PT-N02, and PT-N03. The pharmacokinetic parameters are summarized in Table 12. Compared with the extremely low nasal absorption of PT-N01 (semaglutide dissolved in PBS solution), incorporation of a penetration enhancer, Penetration (PT-N02) or dodecylphosphocholine (DPC), could achieve more rapid nasal absorption in rats and higher plasma concentrations of semaglutide, and at the same time, penetratin or DPC also significantly improved the bioavailability. Table 12. Pharmacokinetic parameters of semaglutide after intranasal administration of PT-N01, PT-N02, and PT-N03 in rats. (n = 3) Example 4. Pharmacokinetics and pharmacodynamics of intranasal or subcutaneous administration of semaglutide in rabbits.

[0057] The purpose of this study was to investigate the pharmacokinetics and pharmacodynamics of semaglutide after intranasal spray and subcutaneous injection. Three groups of white rabbits (n = 3 per dose) were administered a single intranasal dose (4 mg / animal) of three nasal spray compositions (PT-N01, PT-N04, PT-N05), which were prepared according to Example 1 of the present invention, or received a single subcutaneous injection of PT-S01 (0.25 mg / animal) prepared according to Example 2 of the present invention. Multiple blood samples were collected from the ear vein at 0 (before dosing) and 0.5, 1, 2, 4, 6, 8, 12, 24, 48, 72, 96, 144 hours after dosing. The concentration of semaglutide in rabbit plasma was determined using a validated LC / MS / MS method. Pharmacokinetic parameters were generated using standard non-compartmental methods. Blood glucose tests were performed at 0 (before dosing), 0.5, 1, 2, 4, 6, 8, 12, 24, 48, 72, 96, and 144 hours after dosing, while body weight and food intake were recorded once a day.

[0058] Figure 2 Plots of mean semaglutide plasma concentration versus time are shown after single-dose intranasal administration of PT-N01, PT-N04, PT-N05 at 4 mg / animal and single-dose subcutaneous administration of PT-S01 at 0.25 mg / rabbit. Pharmacokinetic parameters are summarized in Table 13. The results showed that intranasal administration of our optimal formulation (PT-N04) could achieve drug plasma concentrations comparable to those of a single subcutaneous dose solution, and the bioavailability of PT-N04 was approximately 28-fold that of the semaglutide PBS solution (PT-N01). In contrast, the enhancement achieved using SNAC as a penetration enhancer (PT-N05) was lower (about 9-fold). Table 13. Pharmacokinetic parameters of semaglutide in rats after intranasal administration of PT-N01, PT-N04, and PT-N05 at a dose of 4 mg / animal or subcutaneous injection of PT-S01. (n = 3)

[0059] Pharmacodynamic parameters including blood glucose, body weight, and food intake are shown as Figures 2 - 4 follows. The body weight of the animals decreased by more than 15% on the second day after single intranasal administration of PT-N04, comparable to that of the subcutaneous injection group of PT-S01; the daily food intake decreased by more than 70% on the second to third days after single intranasal administration of PT-N04, which was also comparable to that of the subcutaneous injection group of PT-S01. During the study, more stable blood glucose levels were observed in the PT-N05 and PT-S01 groups compared to the fluctuating and increasing blood glucose levels in the normal saline group and PT-N01 group. Example 5. Pharmacokinetics of semaglutide in beagle dogs after intranasal and oral administration.

[0060] The objective of this study was to compare the pharmacokinetics of semaglutide after a single intranasal spray of PT-N06 (7 mg) and a single oral tablet (7 mg). The pharmacokinetic parameters are shown in Table 14. Eight beagle dogs (4 males and 4 females) participated in this study. All were fasted overnight before dosing and had unrestricted access to water throughout the study. Food was provided 4 hours after dosing. Group 1 (2 males and 2 females) received a single intranasal dose (7 mg / animal) of nasal spray PT-N06 prepared according to Example 1 of the present invention; Group 2 (2 males and 2 females) received a single oral dose of tablet (7 mg / animal). Multiple blood samples were collected at 0 (before dosing), 0.167, 0.333, 0.5, 1, 2, 4, 8, 12, 24, 48, 72, 96, and 144 hours after dosing. The concentration of semaglutide in dog plasma was determined using a validated LC / MS / MS method. Pharmacokinetic parameters were generated using standard non-compartmental methods. Blood glucose, body weight, and food intake were also recorded during the study. Table 14. Pharmacokinetic parameters of semaglutide after intranasal spray of PT-N06 or oral tablet. (n = 3 - 4) References 1. Knudsen LB, Lau J. The Discovery and Development of Liraglutide and Semaglutide. Front Endocrinol (Lausanne). 2019;10:155. 2. Mahapatra MK, Karuppasamy M·Sahoo BM. Therapeutic Potential of Semaglutide, a Newer GLP 1 Receptor Agonist, in Abating Obesity, Non Alcoholic Steatohepatitis and Neurodegenerative diseases: A Narrative Review. Pharmaceutical Research. 2022;39:1233 - 1248. 3. Merkus FW, Verhoef JC, NG Schipper, E Marttin. Nasal mucociliary clearance as a factor in nasal drug delivery. Adv. Drug Deliv Rev. 1998;5;29(1-2):13-38. 4. Pathak K. Mucoadhesion; A prerequisite or a constraint in nasal drug delivery? Int J Pharm Investig. 2011;1(2):62-63. 5. Derakhshankhah H, Jafari S. Cell penetrating peptides: A concise review with emphasis on biomedical applications. Biomed Pharmacother. 2018;108:1090-1096.

Claims

1. A pharmaceutical composition for intranasal administration, comprising an active ingredient and essential excipients, wherein the active ingredient comprises 0.01% to 20% by weight (w / v) of semaglutide or a pharmaceutically acceptable salt thereof, and the pharmaceutical composition further comprises one or more penetration enhancers selected from cell-penetrating peptides, tight junction modulators, and bioadhesives.

2. The pharmaceutical composition according to claim 1, wherein: the pharmaceutical composition is a liquid solution, which comprises semaglutide or a pharmaceutically acceptable salt thereof, a penetration enhancer, a bioadhesive, a solubilizer, a buffer, a chelating agent, an osmotic pressure regulator, and a preservative.

3. The pharmaceutical composition according to claim 2, wherein: the penetration enhancer is selected from penetratin, dodecylphosphocholine, or dimethyl-β-cyclodextrin.

4. The pharmaceutical composition according to claim 3, wherein: the penetratin accounts for 0.1% to 10% (w / v) of the composition.

5. The pharmaceutical composition according to claim 3, wherein: the dodecylphosphocholine accounts for 0.1% to 10% (w / v) of the composition.

6. The pharmaceutical composition according to claim 3, wherein: the dimethyl-β-cyclodextrin accounts for 0.1% to 50% (w / v) of the composition.

7. The pharmaceutical composition according to claim 2, wherein: the bioadhesive is selected from hydroxypropylmethylcellulose, hydroxypropylcellulose, sodium carboxymethylcellulose, chitosan, sodium hyaluronate, poloxamer 188 / 405, and carbomer 934P.

8. The pharmaceutical composition according to claim 7, wherein: the hydroxypropylmethylcellulose accounts for 0.05% to 5% (w / v) of the composition.

9. The pharmaceutical composition according to claim 7, wherein: the hydroxypropylcellulose accounts for 0.05% to 5% (w / v) of the composition.

10. The pharmaceutical composition according to claim 7, wherein: the sodium carboxymethylcellulose accounts for 0.01% to 10% (w / v) of the composition.

11. The pharmaceutical composition according to claim 7, wherein: the chitosan accounts for 0.01% to 5% (w / v) of the composition.

12. The pharmaceutical composition according to claim 7, wherein: the sodium hyaluronate accounts for 0.01% to 5% (w / v) of the composition.

13. The pharmaceutical composition according to claim 7, wherein: the poloxamer 188 accounts for 0.01% to 5% (w / v) of the composition, and the poloxamer 405 accounts for 0.1% to 30% (w / v) of the composition.

14. The pharmaceutical composition according to claim 7, wherein: the carbomer 934P accounts for 0.05% to 10% (w / v) of the composition.

15. The pharmaceutical composition according to claim 2, wherein: the solubilizer is selected from cyclodextrin or cyclodextrin derivatives.

16. The pharmaceutical composition according to claim 15, wherein: The cyclodextrin or cyclodextrin derivative accounts for 0.05% to 50% (w / v) of the composition.

17. The pharmaceutical composition according to claim 2, wherein: the buffer is selected from sodium dihydrogen phosphate, disodium phosphate, sodium citrate and citric acid.

18. The pharmaceutical composition according to claim 2, wherein: the pharmaceutical composition has a pH in the range of 3.0 to 9.

0.

19. The pharmaceutical composition according to claim 2, wherein: the chelating agent is disodium edetate.

20. The pharmaceutical composition according to claim 19, wherein: the disodium edetate accounts for 0.1% to 5% (w / v) of the composition.

21. The pharmaceutical composition according to claim 2, wherein: the preservative is benzalkonium chloride.

22. The pharmaceutical composition according to claim 21, wherein: the benzalkonium chloride accounts for 0.01% to 0.1% (w / v) of the composition.

23. The pharmaceutical composition according to claim 2, wherein: the osmotic pressure regulator is selected from sodium chloride, mannitol and sorbitol.

24. The pharmaceutical composition according to claim 2, wherein: the liquid solution is formulated as a nasal spray or nasal drops for intranasal administration to a mammal.

25. The pharmaceutical composition according to claim 1, wherein: the intranasal administration device comprises a bottle and a metered multi-dose pump.

26. The pharmaceutical composition according to claim 2, wherein: the pharmaceutical composition is formulated and delivered intranasally in a volume of about 0.05 mL to 0.25 mL of the composition per spray.

27. The pharmaceutical composition according to claim 2, wherein: the intranasal administration is achieved by using a spraying device to deliver a dose of 0.005 mg to 50 mg of semaglutide per spray intranasally.

28. The pharmaceutical composition according to claim 1, wherein: the pharmaceutical composition is formulated as a suspension, emulsion, bioadhesive or in-situ gel, microspheres, nanoparticles or self-emulsifying drug delivery system.

29. A method of treating or preventing diabetes, obesity, non-alcoholic fatty liver disease (NAFLD) or neurodegenerative disease, the method comprising intranasally administering to a mammalian subject in need thereof a liquid solution of the pharmaceutical composition of claim 2 in an effective therapeutic dose.

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