Nasal administration composition containing GLP-1 receptor agonist and preparation method and application thereof

Abstract

The invention relates to a nasal administration composition containing a GLP-1 receptor stimulant and a preparation method and application thereof. In-vivo and in-vitro tests show that the composition has good stability, good bioavailability is achieved after nasal administration of rats, and little drug is distributed in brain tissue.

Description

Composition for nasal administration containing GLP-1 receptor agonist, preparation method and use thereof Technical Field

[0001] The present invention belongs to the field of medical technology, and in particular relates to a composition for nasal administration containing a GLP-1 receptor agonist, and a preparation method and use thereof. Background Art

[0002] Diabetes mellitus (DM) is one of the most common chronic diseases. my country has the highest diabetes prevalence in the world, with an adult diabetes prevalence as high as 11.9%. Type 2 diabetes mellitus (T2DM) accounts for approximately 90%, with a significantly higher prevalence in obese and overweight individuals. T2DM is caused by a combination of pancreatic β-cell dysfunction, impaired insulin action, and abnormal glucagon secretion. Glucagon-like peptide-1 (GLP-1) receptor agonists are a new class of glucose-lowering drugs for the treatment of T2DM. They significantly ameliorate key pathophysiological deficits in T2DM and offer additional benefits beyond glucose control, such as reduced cardiovascular mortality, improved atherosclerosis, reduced weight, lowered systolic blood pressure, and improved lipid profiles, offering new hope for patients with T2DM. The development of GLP-1 receptor agonists has evolved from short-acting (two to three daily injections) to long-acting (once daily) to ultra-long-acting (once weekly). Currently, all GLP-1 receptor agonists marketed globally are administered systemically, most of which are administered by injection, among which semaglutide can be administered orally.

[0003] Although the administration routes of GLP-1 receptor agonists have been greatly improved, the inherent defects of systemic administration still cause great inconvenience to diabetic or overweight / obese patients who need long-term medication, so there is still a need to develop administration routes that are more in line with patient compliance.

[0004] Nasal administration has the advantages of rapid absorption, fast onset of action, avoidance of first-pass effect in the liver, high bioavailability, ease of use, and the ability to bypass the blood-brain barrier and deliver drugs directly to the brain. Although a variety of nasal peptide products have been successfully marketed or entered clinical trials, nasal absorption of drugs is a very complex process. For example, Huang Zhuang et al. pointed out in the article "Research Progress on Nasal Administration of Peptide Drugs" that factors affecting nasal absorption of drugs include but are not limited to: drug properties (such as relative molecular weight, lipophilicity), formulation characteristics (e.g., formulation pH, osmotic pressure, viscosity), and nasal environment (e.g., enzyme activity, ciliary clearance, nasal mucus). Therefore, whether a specific nasal absorption enhancer can achieve nasal absorption of a certain peptide is highly unpredictable.

[0005] CN116942612A discloses a method for achieving nasal spray absorption of GLP-1 receptor agonists such as semaglutide using caprylic acid capric acid macrogol glyceride. However, the content of the main functional delivery agent caprylic acid capric acid macrogol glyceride is as high as 5%, and special drug delivery equipment is usually required to achieve spray delivery of the drug.

[0006] Although brain targeting is one of the potential advantages of nasal administration, for example, US2021087250A points out that nasal administration of peptide drugs usually has brain targeting as one of the main purposes. In addition, US2011129462A1 has proved through experiments that nasal absorption enhancers such as DDM can achieve brain-targeted delivery of peptide compounds such as antibody fragments. The instructions for the FDA-approved hypoglycemic version of semaglutide (trade name Ozempic) point out that the hypoglycemic effect of semaglutide mainly depends on its regulation of insulin secretion, so it does not need to enter the CNS to exert its hypoglycemic effect; although the instructions for the FDA-approved weight-loss version of semaglutide (trade name Wegovy) point out that semaglutide may exert a weight-loss effect by activating the GLP-1 receptor that regulates appetite in the CNS, it does not rule out the possibility that other mechanisms of action are involved (such as delaying gastric emptying, etc.). Taking into account the adverse reactions that may affect the risk-benefit profile after the drug enters the central nervous system, for example, Jia-Rui Li et al. reported two cases of adverse reactions of depression caused by the use of semaglutide in the article Semaglutide-associated depression: a report of two cases. The authors attributed these adverse reactions to semaglutide stimulating GLP-1 receptors located in the central nervous system. Therefore, avoiding or reducing the brain targeting of GLP-1 receptor agonist drugs will help improve their risk-benefit profile.

[0007] There is a need in the art for a technical solution to achieve nasal administration of GLP-1 receptor agonists, including semaglutide, and reduce / avoid their brain-targeted delivery.

[0008] SUMMARY OF THE INVENTION

[0009] One of the objects of the present invention is to provide a composition containing a GLP-1 receptor agonist for nasal administration, which can also avoid targeting the central nervous system.

[0010] In order to achieve the above object, the present invention provides a composition containing a GLP-1 receptor agonist and a nasal absorption enhancer, wherein the GLP-1 receptor agonist is one selected from liraglutide, semaglutide and tilpotide; the nasal absorption enhancer is one selected from dodecyl-β-D-maltoside, nonyl-β-D-pyranoside, polyoxyethylene (7) lauryl ether, polyoxyethylene (8) lauryl ether, polyoxyethylene (9) lauryl ether, polyethylene glycol One or more of octylphenyl alcohol ether, sodium lauryl sulfate, polysorbate 80, diethylene glycol monoethyl ether, polyethylene glycol 400, polysorbate 20, ethoxylated C16-18-alcohol, polyoxyethylene (10) hexadecyl ether, polyoxyethylene (10) stearyl ether, polyoxyethylene (10) oleyl ether, polyoxyethylene (20) oleyl ether, n-octyl-β-D-pyranoglucoside, 1-O-decyl-β-D-maltoside and polyoxyethylene (2) hexadecyl ether.

[0011] Another object of the present invention is to provide a method for preparing the composition of the present invention, wherein the method comprises mixing a GLP-1 receptor agonist with a nasal absorption enhancer.

[0012] Another object of the present invention is to provide a use of the composition of the present invention in the preparation of a medicament for treating a disease, wherein the disease is one or more selected from type 2 diabetes, overweight or obesity.

[0013] Both in vivo and in vitro tests show that the composition of the present invention has good stability, good bioavailability after nasal administration to rats, and very little drug distribution in brain tissue. BRIEF DESCRIPTION OF THE DRAWINGS

[0014] Figure 1: HPLC chart of formulation C1 in Example 3 measured at 50°C for 10 days;

[0015] Figure 2: HPLC chart of formulation C2 in Example 3 measured at 50°C for 10 days;

[0016] Figure 3: HPLC chart of formulation C3 in Example 3 measured at 50°C for 10 days;

[0017] Figure 4: HPLC chart of formulation C4 in Example 3 measured at 50°C for 10 days;

[0018] Detailed Description of the Invention

[0019] In a first aspect, the present invention provides a composition containing a GLP-1 receptor agonist and a nasal absorption enhancer, wherein the GLP-1 receptor agonist is one selected from liraglutide, semaglutide and tilpotide; the nasal absorption enhancer is one selected from dodecyl-β-D-maltoside, nonyl-β-D-pyranoglucoside, polyoxyethylene (7) lauryl ether, polyoxyethylene (8) lauryl ether, polyoxyethylene (9) lauryl ether, polyethylene glycol octyldodecyl ether, polyoxyethylene (10) lauryl ether, polyoxyethylene (11) lauryl ether, polyoxyethylene (12) lauryl ether, polyoxyethylene (13) lauryl ether, polyoxyethylene (14) lauryl ether, polyoxyethylene (15) lauryl ether, polyoxyethylene (16) lauryl ether, polyoxyethylene (17) lauryl ether, polyoxyethylene (18) lauryl ether, polyoxyethylene (19) lauryl ether, polyoxyethylene (20) lauryl ether, polyoxyethylene (21) lauryl ether, polyoxyethylene (22) lauryl ether, polyoxyethylene (23) lauryl ether, polyoxyethylene (24) lauryl ether, polyoxyethylene (25) lauryl ether, polyoxyethylene (26) lauryl ether, polyoxyethylene (27) lauryl ether, polyoxyethylene (28) lauryl ether, polyoxyethylene (29) lauryl ether, polyoxyethylene (30) lauryl ether, polyoxyethylene (31) lauryl ether, polyoxyethylene (32) lauryl ether, polyoxyethylene (33) lauryl ether, polyoxyethylene (34) lauryl ether, polyoxyethylene (35) lauryl ether, polyoxyethylene (36) lauryl ether, polyoxyethylene (37) lauryl ether, poly One or more of alkylphenyl ether, sodium lauryl sulfate, polysorbate 80, diethylene glycol monoethyl ether, polyethylene glycol 400, polysorbate 20, ethoxylated C16-18-alcohol, polyoxyethylene (10) hexadecyl ether, polyoxyethylene (10) stearyl ether, polyoxyethylene (10) oleyl ether, polyoxyethylene (20) oleyl ether, n-octyl-β-D-pyranoglucoside, 1-O-decyl-β-D-maltoside and polyoxyethylene (2) hexadecyl ether.

[0020] According to some embodiments of the invention, the composition is liquid.

[0021] According to some embodiments of the present invention, the composition uses water as a solvent.

[0022] According to some embodiments of the invention, the GLP-1 receptor agonist is semaglutide.

[0023] According to some embodiments of the present invention, the nasal absorption enhancer is selected from the group consisting of dodecyl-β-D-maltoside and maltoside having the molecular formula CH3(CH2) n-1 [OCH2CH2] m One or more OH-type absorption enhancers, wherein n and m are both integers. In some embodiments, n is an integer selected from 10-16; m is an integer selected from 4-10.

[0024] According to some embodiments of the present invention, the nasal absorption enhancer is selected from the group consisting of dodecyl-β-D-maltoside, polyoxyethylene (7) lauryl ether, polyoxyethylene (8) lauryl ether, and polyoxyethylene (9) lauryl ether. In some embodiments, the nasal absorption enhancer is dodecyl-β-D-maltoside or polyoxyethylene (9) lauryl ether.

[0025] According to some embodiments of the present invention, the content of the GLP-1 receptor agonist in the composition is 0.28 mg / mL to 170.0 mg / mL. In some embodiments, the content of the GLP-1 receptor agonist in the composition is 17.0 mg / mL to 70.0 mg / mL. In some embodiments, the content of the GLP-1 receptor agonist in the composition is 5.25 mg / mL to 22.2 mg / mL. In some embodiments, the content of the GLP-1 receptor agonist in the composition is 5.25 mg / mL to 10.5 mg / mL.

[0026] According to some embodiments of the present invention, the w / v% content of the nasal absorption enhancer in the composition is 0.25% to 2.5%. In some embodiments, the w / v% content of the nasal absorption enhancer in the composition is 0.25% to 1.0%. In some embodiments, the w / v% content of the nasal absorption enhancer in the composition is 0.25% to 0.5%.

[0027] According to some embodiments of the present invention, the mass ratio of the nasal absorption enhancer to the GLP-1 receptor agonist in the composition is 0.01 to 18. In some embodiments, the mass ratio of the nasal absorption enhancer to the GLP-1 receptor agonist in the composition is 0.03 to 1.50.

[0028] According to some embodiments of the present invention, the composition further comprises a preservative. In some embodiments, the preservative is selected from phenol, benzalkonium chloride, benzyl ammonium chloride, 2-trichloromethyl-2-propanol, butyl p-hydroxybenzoate, propyl p-hydroxybenzoate, benzethonium chloride, chlorocresol, and benzoic acid.

[0029] In another aspect, the present invention provides a method for preparing the composition of the present invention, wherein the method comprises mixing a GLP-1 receptor agonist with a nasal absorption enhancer.

[0030] In another aspect, the present invention provides use of the composition of the present invention in the preparation of a medicament for treating a disease, wherein the disease is one or more selected from type 2 diabetes, overweight or obesity. Example

[0031] Example 1: PAMPA (parallel artificial membrane permeability assay) test of the test formulation

[0032] 1.1. Objective: To investigate the in vitro permeability differences of 10 mM semaglutide in Prisma HT buffer, pH 7.4, with the addition of different types and ratios of permeation enhancers using the PAMPA method.

[0033] 1.2. Test methods

[0034] (1) Prescription 1-20 composition

[0035] An appropriate amount of 10 mM semaglutide stock solution was measured and placed in a liquid injection vial. Different penetration enhancers were weighed and added according to the types and proportions listed in Table 1. A magnetic stirrer was added, and the magnetic stirrer was turned on and the speed was adjusted to 650 rpm. The temperature of the magnetic stirrer was maintained at 37°C and stirred for later use.

[0036] Table 1

[0037] (2) PAMPA test

[0038] The supply end solution is the prescription 1-20 solution;

[0039] The receiving end solution is pH 7.4 Prisma HT buffer

[0040] ① Hydration of the PAMPA membrane: Prepare the Hydration Solution (Pion) (use at room temperature). Carefully remove the donor end of the PAMPA plate and add 200 μl of the solution to each well. Observe whether the surface of each well forms a curved surface due to surface tension. Carefully cover the receiving end of the PAMPA plate, minimizing the formation of air bubbles on the contact surface. Allow the membrane to remain moist and hydrate overnight. The hydrated membrane will appear translucent or have white spots. The PAMPA experiment can now begin.

[0041] ②Perform the operation on a PAMPA plate, with four replicates for each recipe, and add 200 μl of the corresponding supply end solution to the bottom of each plate.

[0042] ③ Gently cover the membrane on the supply end solution to avoid bubbles in the middle (if too many bubbles are generated, put it in a vacuum drying oven at 25℃ and vacuum it 2-3 times).

[0043] ④ Add 200 μl of the corresponding receiving end solution to the receiving end respectively. Be careful to add the solution to the membrane to avoid forming a cavity.

[0044] ⑤ After incubation at 37°C for 5 hours, take the supply solution and receiving solution and perform sample processing.

[0045] (3) Sample processing

[0046] Take 100 μl of each of the donor, receiver, and 0 h samples and dilute with 900 μl of 50% acetonitrile. Centrifuge at 13,000 rpm, 25°C, and 5 min. Transfer 200 μl of the supernatant to a liquid chromatography injection vial and submit for analysis by HPLC. The HPLC parameters are shown in Table 2.

[0047] Table 2

[0048] (4) Calculation formula

[0049] VA – the volume in the acceptor wells (ml);

[0050] VD – the volume in the donor wells (ml);

[0051] A – the filtration area (cm 2 ));

[0052] t – incubation time (s);

[0053] tLAG – the steady-state time (s);

[0054] CD(t) – the concentration of the compound in the donor well at time t (lM);

[0055] CA(t) – the concentration of the compound in the acceptor well at time t (lM);

[0056] CD(0) – the concentration of the compound in donor well at time 0 (lM).

[0057] 1.3. PAMPA test results statistics

[0058] As shown in Table 3.

[0059] Table 3

[0060] Among the parameters in the above table, the smaller the supply percentage, the larger the penetration percentage, the larger the LogPe value (closer to a positive value), and the larger the Pe value, the greater the penetration amount, and relatively speaking, the better the permeability of the prescription.

[0061] Conclusion

[0062] According to the in vitro permeation data of Skin PAMPA, at the same concentration of 0.25% (w / v%) permeation enhancer, dodecyl-β-D-maltoside (DDM), polyoxyethylene (8) lauryl ether (C12E8), and polysorbate 20 (Tween 20) have better permeation enhancement effects, and the permeation enhancement effect is improved by appropriately increasing the permeation enhancer dosage, such as increasing it to 0.50% (w / v%).

[0063] From the PAMPA experiment, a penetration enhancer with better permeation effect was selected and prepared into a nasal drop preparation. After nasal administration to SPF rats, the drug concentration in the rat plasma and brain tissue was measured over time.

[0064] Example 2. Pharmacokinetics - Determination of bioavailability

[0065] 2.1. Test materials and test substances

[0066] (1) Animals: SD rats (SPF grade), 7-8 weeks old, female weighing 210-230 g, male weighing 260-290 g. 4 rats per group, half male and half female.

[0067] (2) Test prescription: as shown in Table 4

[0068] Table 4 Note: a Each prescription contains 0.55g / 100ml of phenol as an antibacterial agent b The concentration of each nasal absorption enhancer is w / v% relative to the entire formulation

[0069] 2.2. Methods

[0070] Administration: All drugs were administered in a single dose. The administration route and dosage are shown in Table 4.

[0071] Sampling: Blood samples were collected before dosing (0 h) and at 0.083, 0.5, 0.75, 1, 2, 4, 6, 8, 12, and 24 h after dosing. Approximately 0.20 mL of blood was collected from the retroorbital venous plexus and anticoagulated with EDTA. Blood samples were placed on ice and centrifuged within 2 h to separate plasma (centrifugation conditions: 3500 rpm, 10 min, 2-8°C).

[0072] Determination conditions: The HPLC-MS parameters are shown in Table 5.

[0073] Table 5

[0074] Results

[0075] The main pharmacokinetic parameters of each prescription are shown in Table 6.

[0076] Table 6 *Note: “Relative bioavailability (F)” is defined as: F = (AUCT*DR) / (AUCR*DT)*100% AUCT: Area under the plasma concentration-time curve of nasal drops; DR: Subcutaneous dose; AUCR: Area under the plasma concentration-time curve of subcutaneous dose; DT: Subcutaneous dose

[0077] The above animal experiment results show that different concentrations of dodecyl-β-D-maltoside (DDM) and polyoxyethylene (9) dodecyl ether (C12E9) can be well absorbed into the blood when combined with different concentrations of semaglutide.

[0078] Example 3. Compatibility / stability studies

[0079] The HPLC conditions in Example 1 were used to determine the changes in semaglutide content and impurities in semaglutide solution, semaglutide plus different penetration enhancer solutions, and semaglutide plus different antibacterial agent solutions stored in the dark at 40° C. and 50° C. for 10 days (10 D). The results are shown in Table 7.

[0080] Table 7

[0081] Note: SEM is semaglutide. All prescriptions are aqueous solutions, and the concentration of SEM is 5.252 mg / ml.

[0082] The definitions of hydrophilic impurities 1 and 2 and hydrophobic impurities described in Table 7 are shown in the HPLC charts of formulations C1 to C4 measured at 50° C. for 10 days as shown in Figures 1 to 4 .

[0083] As a control, the main peak intensities of compositions J and K of CN116942612A were measured at 50° C. for 5 days using the same method, and both were found to be lower than 95%.

[0084] Results: Both penetration enhancers had a protective effect on semaglutide and inhibited its degradation. Benzalkonium chloride, as an antibacterial agent in nasal spray preparations, also had a protective effect on semaglutide in this product.

[0085] Example 4. Pharmacokinetics Test - Brain Tissue Distribution

[0086] 4.1. Test materials and test substances

[0087] (1) Animals: Same as Example 2. Each test prescription group consisted of 40 rats.

[0088] (2) Tested prescriptions: F5, F8 and F23 in Example 2.

[0089] 4.2. Methods

[0090] Administration: Same as Example 2.

[0091] Sampling: Five rats of both sexes were sacrificed by dislocation at 1, 8, 16, and 24 hours after administration. The brains were removed, quickly rinsed with clean water, and accurately weighed. The tissue was minced and mixed with pre-chilled saline at a ratio of 1 g:3 mL. The mixture was homogenized using an ultrasonic cell disruptor and centrifuged at 12,000 rpm for 2 minutes. The supernatant was stored at -20°C for testing. The assay concentration of semaglutide in the tissue homogenate (ng / mL) was divided by the tissue homogenate density (g tissue / mL) to calculate the tissue concentration in ng / g tissue.

[0092] Detection: Conditions are the same as in Example 2.

[0093] Results

[0094] As shown in Table 8.

[0095] Table 8 BLQ = below the detection limit

[0096] As shown in Table 8, the distribution of F5 and F8 in the rat brain after nasal administration was significantly lower than that of F23 after injection.

[0097] Example 5. Pharmacokinetics - Brain Tissue Distribution

[0098] The method is the same as that of Example 4, except that the types of prescriptions tested are increased, i.e., prescriptions F6, F7, F9, F10, F13, and F14 selected from Table 6 are tested.

[0099] Prescriptions F6, F7, F9, and F10 could detect semaglutide in brain tissue at at least one time point. Among them, the semaglutide doses of F6, F7, and F10 were the same as those of F5 and F8, but the types of nasal absorption enhancers were different, indicating that there is great uncertainty as to whether nasal absorption enhancers can avoid brain delivery.

[0100] In addition, no drug was detected in brain analysis of the highest doses of F13 and F14 at any time point.

Claims

1. A composition containing a GLP-1 receptor agonist and a nasal absorption enhancer, characterized in that: The GLP-1 receptor agonist is one selected from liraglutide, semaglutide and tilpotide; the nasal absorption enhancer is one selected from dodecyl-β-D-maltoside, nonyl-β-D-pyranoglucoside, polyoxyethylene (7) dodecyl ether, polyoxyethylene (8) dodecyl ether, polyoxyethylene (9) dodecyl ether, polyethylene glycol octylphenyl ether, sodium dodecyl sulfate, polysorbate 80, diethylene glycol monoethyl ether, polyethylene glycol 400, polysorbate 20, ethoxylated C16-18-alcohol, polyoxyethylene (10) hexadecyl ether, polyoxyethylene (10) octadecyl ether, polyoxyethylene (10) oleyl ether, polyoxyethylene (20) oleyl ether, n-octyl-β-D-pyranoglucoside, 1-O-decyl-β-D-maltoside and polyoxyethylene (2) hexadecyl ether.

2. The composition according to claim 1, characterized in that The composition is liquid.

3. The composition according to claim 2, characterized in that The composition uses water as solvent.

4. The composition according to claim 1, characterized in that The GLP-1 receptor agonist is semaglutide.

5. The composition according to claim 1, characterized in that The nasal absorption enhancer is selected from dodecyl-β-D-maltoside and CH3(CH2) n-1 [OCH2CH2] m One or more OH-type absorption enhancers, wherein n and m are both integers.

6. The composition according to claim 5, characterized in that n is an integer selected from 10-16; m is an integer selected from 4-10.

7. The composition according to claim 1, characterized in that The nasal absorption enhancer is selected from dodecyl-β-D-maltoside, polyoxyethylene (7) dodecyl ether, polyoxyethylene (8) dodecyl ether and polyoxyethylene (9) dodecyl ether, preferably dodecyl-β-D-maltoside or polyoxyethylene (9) dodecyl ether.

8. The composition according to claim 1, characterized in that The content of the GLP-1 receptor agonist in the composition is 0.28 mg / mL to 170.0 mg / mL.

9. The composition according to claim 8, characterized in that The content of the GLP-1 receptor agonist in the composition is 17.0 mg / mL to 70.0 mg / mL.

10. The composition according to claim 8, characterized in that The content of the GLP-1 receptor agonist in the composition is 5.25 mg / mL to 22.2 mg / mL, preferably 5.25 mg / mL to 10.5 mg / mL.

11. The composition according to claim 1, characterized in that The w / v% content of the nasal absorption enhancer in the composition is 0.25% to 2.5%.

12. The composition according to claim 11, characterized in that The w / v% content of the nasal absorption enhancer in the composition is 0.25% to 1.0%, preferably 0.25% to 0.5%.

13. The composition according to claim 1, characterized in that The mass ratio of the nasal absorption enhancer to the GLP-1 receptor agonist in the composition is 0.03 to 1.

50.

14. The composition according to claim 1, characterized in that The composition may further contain a preservative, such as phenol, benzalkonium chloride, benzyl ammonium chloride, 2-trichloromethyl-2-propanol, butyl p-hydroxybenzoate, propyl p-hydroxybenzoate, benzethonium chloride, chlorocresol, benzoic acid, preferably benzalkonium chloride.

15. Use of the composition according to any one of claims 1 to 14 in the preparation of a medicament for treating a disease, characterized in that: The disease is one or more selected from type 2 diabetes, overweight or obesity.

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