Nasal compositions containing glp-1 receptor agonists and methods of making and using the same

Abstract

The application belongs to the technical field of medicine, and particularly relates to a composition for nasal administration containing a GLP-1 receptor agonist and a preparation method and use thereof. In-vivo and in-vitro tests show that the composition has good stability, has good bioavailability after nasal administration in rats, and has little drug distribution in brain tissue.

Description

Technical Field

[0001] This invention belongs to the field of pharmaceutical technology, specifically relating to nasal-administered compositions containing GLP-1 receptor agonists, their preparation methods, and uses. Background Technology

[0002] Diabetes mellitus (DM) is one of the most common chronic diseases. my country has the largest number of diabetes patients in the world, with a prevalence rate of 11.9% among adults. Type 2 diabetes mellitus (T2DM) accounts for approximately 90% of these cases, and the prevalence is significantly higher in obese and overweight individuals. T2DM is caused by the combined effects of pancreatic β-cell dysfunction, insulin resistance defects, and abnormal glucagon secretion. Glucagon-like peptide-1 (GLP-1) receptor agonists are a new class of hypoglycemic agents for the treatment of T2DM. They can significantly improve some key pathophysiological defects in T2DM and offer additional benefits beyond just lowering blood sugar, such as reducing cardiovascular mortality, improving atherosclerosis, reducing body weight, lowering systolic blood pressure, and improving lipid profiles, bringing new hope to patients with T2DM. The development of GLP-1 receptor agonists has progressed from short-acting (2-3 injections daily) to long-acting (1 injection daily) and then to ultra-long-acting (1 injection weekly). Currently, all GLP-1 receptor agonists available globally are systemic administrations, with most being administered by injection. However, semaglutide has achieved oral administration.

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

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

[0005] CN116942612A discloses a method for nasal spray absorption of GLP-1 receptor agonists such as smegglutide using polyethylene glycol glycerol octanoate and caprylic acid. However, the main functional delivery agent, polyethylene glycol glycerol octanoate, has a content as high as 5%, and special drug delivery devices are usually required to achieve spray drug delivery.

[0006] While brain targeting is one of the potential advantages of nasal administration—for example, US2021087250A states that nasal administration of peptide drugs is often aimed at achieving brain targeting—US2011129462A1 has demonstrated through experiments that nasal absorption enhancers such as DDM can achieve brain-targeted delivery of peptide compounds such as antibody fragments. The FDA-approved hypoglycemic version of semaglutide (trade name Ozempic) states that its hypoglycemic effect mainly relies on its regulation of insulin secretion, thus it does not need to enter the CNS to exert its hypoglycemic effect. Although the FDA-approved weight-loss version of semaglutide (trade name Wegovy) states that semaglutide may exert its weight-loss effect by activating GLP-1 receptors in the CNS that regulate appetite, it does not rule out the possibility of other mechanisms of action (such as delaying gastric emptying). Considering the potential adverse reactions and risk-benefit profile of drugs after entering the central nervous system, such as those observed by Jia-Rui Li et al. Semaglutide-associated depression: a report of two cases The article reported two cases of adverse reactions to depression caused by the use of semaglutide. The authors attributed this adverse reaction to the activation of GLP-1 receptors in the central nervous system by semaglutide. Therefore, avoiding or reducing the brain targeting of GLP-1 receptor agonists may help improve their risk-benefit profile.

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

[0008] One of the objectives of this invention is to provide a composition containing a GLP-1 receptor agonist for nasal administration, wherein the composition also avoids delivery targeting the central nervous system.

[0009] To achieve the above objectives, the present invention provides a composition containing a GLP-1 receptor agonist and a nasal absorption enhancer, wherein the GLP-1 receptor agonist is selected from liraglutide, smegglutide, and telposide; and the nasal absorption enhancer is selected from dodecyl-β-D-maltodextrin, nonyl-β-D-glucopyranoside, 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-ol, polyoxyethylene (10) hexadecyl ether, polyoxyethylene (10) octadecyl ether, polyoxyethylene (10) oil ether, and polyoxyethylene (20) oil ether. One or more of n-octyl-β-D-glucopyranoside, 1-O-decyl-β-D-maltodextrin, and polyoxyethylene (2) hexadecyl ether.

[0010] 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.

[0011] Another object of the present invention is to provide the use of the compositions of the present invention in the preparation of a medicament for treating a disease, wherein the disease is selected from one or more of type 2 diabetes, overweight or obesity.

[0012] Both in vivo and in vitro tests showed that the composition of the present invention has good stability, good bioavailability after intranasal administration to rats, and minimal drug distribution in brain tissue. Attached Figure Description

[0013] Figure 1 Example 3: HPLC chromatogram of formulation C1 measured at 50°C for 10 days; Figure 2 Example 3: HPLC chromatogram of formulation C2 measured at 50°C for 10 days; Figure 3 Example 3: HPLC chromatogram of formulation C3 measured at 50°C for 10 days; Figure 4 Example 3: HPLC chromatogram of formulation C4 measured at 50°C for 10 days; Invention Details

[0014] In a first aspect, the present invention provides a composition comprising a GLP-1 receptor agonist and a nasal absorption enhancer, wherein the GLP-1 receptor agonist is selected from liraglutide, smegglutide, and telposide; and the nasal absorption enhancer is selected from dodecyl-β-D-maltodextrin, nonyl-β-D-glucopyranoside, polyoxyethylene (7) dodecyl ether, polyoxyethylene (8) dodecyl ether, polyoxyethylene (9) dodecyl ether, and polyethylene glycol octyl ether. One or more of the following: phenyl ether, sodium dodecyl sulfate, polysorbate-80, diethylene glycol monoethyl ether, polyethylene glycol 400, polysorbate-20, ethoxylated C16-18-ol, polyoxyethylene (10) hexadecyl ether, polyoxyethylene (10) octadecyl ether, polyoxyethylene (10) oil ether, polyoxyethylene (20) oil ether, n-octyl-β-D-glucopyranoside, 1-O-decyl-β-D-maltodextrin and polyoxyethylene (2) hexadecyl ether.

[0015] According to some embodiments of the present invention, the composition is liquid.

[0016] According to some embodiments of the present invention, the composition uses water as a solvent.

[0017] According to some embodiments of the present invention, the GLP-1 receptor agonist is smegglutinin.

[0018] According to some embodiments of the present invention, the nasal absorption enhancer is selected from dodecyl-β-D-maltodextrin and compounds with the molecular formula CH3(CH2). n-1 [OCH2CH2] m One or more OH-type absorption promoters, wherein n and m are both integers. In some embodiments, n is an integer selected from 10 to 16; m is an integer selected from 4 to 10.

[0019] According to some embodiments of the present invention, the nasal absorption enhancer is selected from dodecyl-β-D-maltose, polyoxyethylene (7) dodecyl ether, polyoxyethylene (8) dodecyl ether, and polyoxyethylene (9) dodecyl ether. In some embodiments, the nasal absorption enhancer is dodecyl-β-D-maltose or polyoxyethylene (9) dodecyl ether.

[0020] 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.

[0021] 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%.

[0022] 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.

[0023] 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, butylparaben, propylparaben, benzyl chloride, chlorocresol, and benzoic acid.

[0024] On the other hand, the present invention provides a method for preparing the composition of the present invention, wherein the method includes mixing a GLP-1 receptor agonist with a nasal absorption enhancer.

[0025] In another aspect, the present invention provides the use of the compositions of the present invention in the preparation of a medicament for treating a disease, wherein the disease is selected from one or more of type 2 diabetes, overweight or obesity.

[0026] Example

[0027] 1.1. Experimental objective: To investigate the differences in in vitro permeability of 10 mM semaglutide in pH 7.4 PrismaHT buffer with the addition of different types and proportions of permeation enhancers using the PAMPA method.

[0028] 1.2. Test Methods

[0029] Measure an appropriate amount of 10mM smegglutide stock solution into a liquid chromatography vial. Weigh out the corresponding types and proportions of different promoters listed in Table 1, add a magnetic stir bar, turn on the magnetic stirrer and adjust the speed to 650 rpm. Keep the temperature of the magnetic stirrer at 37℃ and stir for later use.

[0030] Table 1

[0031] (2) PAMPA test The solution supplied is the formula 1-20 solution; The receiving solution is a pH 7.4 Prisma HT buffer. ① PAMPA Membrane Hydration: Prepare the hydration solution (Pion) (for use at room temperature). Carefully remove the supply end of the PAMPA plate and add 200 μl of hydration solution to each well. Observe whether each empty surface forms a curved surface due to surface tension. Carefully cover the receiving end of the PAMPA plate, avoiding air bubbles at the contact surface as much as possible. Keep it moist and allow it to hydrate overnight. The hydrated membrane will be translucent or have white spots. At this point, PAMPA experiments can begin.

[0032] ② Perform the operation on a PAMPA plate, with four replicates for each prescription, and add 200 μl of the corresponding supply end solution to the bottom of each.

[0033] ③ Gently cover the solution at the supply end with the membrane to avoid the formation of air bubbles in the middle (if a lot of air bubbles are generated, the membrane can be placed in a vacuum drying oven at 25°C and vacuumed 2-3 times).

[0034] ④ Add 200 μl of the corresponding receiver solution to the receiving end, being careful to add the solution onto the membrane to avoid forming cavities.

[0035] ⑤ After incubating at 37°C for 5 hours, take the supply solution and receiving solution and process the sample.

[0036] Take 100 μl each of the supply end, receiving end, and 0h sample, dilute with 900 μl of 50% acetonitrile, and then centrifuge at 13000 rpm for 5 min at 25°C. Take 200 μl of the supernatant and place it in a HPLC vial for analysis. The HPLC conditions are shown in Table 2.

[0037] Table 2

[0038] (4) Calculation formula

[0039] VA – Receiver volume (the volume in the acceptor wells (ml)); VD – Supply volume (the volume in the donor wells (ml)); A – Filtration area (cm²) 2 )); t – Incubation time (s); tLAG – steady-state time (s); CD(t) – The concentration of the compound in the feed well at time t (lM); CA(t) – The concentration of the compound in the acceptor well at time t (lM); CD(0) – The concentration of the compound indonor well at time 0 (lM) in the feed end.

[0040] As shown in Table 3.

[0041] Table 3 +0.25% Dodecyl-β-D-maltodextrin (DDM) 4.1613 3.860 0.034 92.8 0.8 -6.45 0.35 +0.25% Nonyl-β-D-glucopyranoside (NG) 4.1253 3.806 0.008 92.3 0.2 -7.06 0.09 +0.25% Polyoxyethylene (8) Dodecyl Ether (C12E8) 3.9326 3.585 0.041 91.2 1.0 -6.35 0.45 +0.50% Polyoxyethylene (8) Dodecyl Ether (C12E8) 4.0167 3.333 0.058 83.0 1.5 -6.16 0.69 +1.00% Polyoxyethylene (8) Dodecyl Ether (C12E8) 4.0557 3.480 0.037 85.8 0.9 -6.37 0.42 +0.25% polyethylene glycol octylphenyl ether (Triton X-100) 3.9121 3.556 0.017 90.9 0.4 -6.73 0.19 +0.25% Sodium Dodecyl Sulfate (SDS) 4.0913 3.669 0.009 89.7 0.2 -7.03 0.09 +0.25% Polysorbate-80 (Tween 80) 3.8413 3.635 0.008 94.6 0.2 -7.07 0.08 +0.25% Diethylene Glycol Monoethyl Ether (Transcutol HP) 4.0353 3.766 0.001 93.3 0.0 -7.87 0.01 +0.25% Polyethylene Glycol 400 (PEG400) 3.9531 3.686 0.003 93.2 0.1 -7.48 0.03 +0.25% Polysorbate-20 (Tween 20) 3.8963 3.603 0.035 92.5 0.9 -6.42 0.38 +0.25% ethoxylated C16-18-ol 3.7370 3.472 0.002 92.9 0.1 -7.60 0.03 +0.25% Polyoxyethylene (10) hexadecyl ether (Brij56) 4.0480 3.736 0.028 92.3 0.7 -6.53 0.29 +0.25% Polyoxyethylene (10) Octadecyl Ether (C18E10) 4.0810 3.701 0.017 90.7 0.4 -6.73 0.18 +0.25% Polyoxyethylene (10) Oil Ether (Brij O10) 3.9486 3.582 0.019 90.7 0.5 -6.65 0.22 +0.25% Polyoxyethylene (20) Oil Ether (Brij O2O) 4.0910 4.062 0.004 99.3 0.1 -7.36 0.04 +0.25% n-Octyl-β-D-glucopyranoside (OGP) 3.9294 3.670 0.003 93.4 0.1 -7.44 0.04 +0.25% 1-O-decyl-β-D-maltodextrin (DM) 4.0348 4.051 0.001 100.4 0.0 -7.84 0.01 +0.25% Polyoxyethylene (9) dodecyl ether (C12E9) 3.8997 3.535 0.024 90.7 0.6 -6.56 0.27 +0.25% Polyoxyethylene (2) hexadecyl ether (Brij 52) 4.0969 3.818 0.006 93.2 0.1 -7.21 0.06 +0.5% DDM 4.0514 3.788 0.036 93.5 0.9 -5.75 0.42 +1% DDM 3.9985 3.826 0.034 95.7 0.6 -7.52 0.15 +0.5% C12E9 4.0004 3.572 0.032 89.3 0.8 -5.45 0.31 +1% C12E9 3.9952 3.767 0.029 94.3 0.5 -7.85 0.11 Among the parameters in the table above, the smaller the supply percentage, the larger the permeation percentage, the larger the LogPe value (closer to a positive value), and the larger the Pe value, the greater the permeation, indicating that the prescription has better permeability.

[0042] 1.4. Conclusion According to Skin PAMPA in vitro permeation data, at the same concentration of 0.25% (w / v%) of permeation enhancer, dodecyl-β-D-maltodextrin (DDM), polyoxyethylene (8) dodecyl ether (C12E8), and polysorbate-20 (Tween 20) showed better permeation enhancement effects. Furthermore, appropriately increasing the amount of permeation enhancer, such as to 0.50% (w / v%), improved the permeation enhancement effect.

[0043] A permeation enhancer with good effect was selected from the PAMPA experiment and formulated into a nasal drop preparation. After nasal administration to SPF-grade rats, the drug concentration in rat plasma and brain tissue was measured over time.

[0044] Example 2. Pharmacokinetic Study - Determination of Bioavailability

[0045] (1) Animals: SD rats (SPF grade), 7-8 weeks old, females weighing 210-230g and males weighing 260-290g. There were 4 rats in each group, half male and half female.

[0046] (2) Test prescriptions: as shown in Table 4 Table 4 Note: a Each prescription contains 0.55g / 100ml of phenol as an antibacterial agent. b The concentrations of each nasal absorption enhancer are all w / v% relative to the entire prescription.

[0047] Administration: All medications were administered as a single dose. The route of administration and dosage are shown in Table 4.

[0048] sampling: Blood samples were collected before administration (0 h) and at 0.083, 0.5, 0.75, 1, 2, 4, 6, 8, 12, and 24 h after administration. Blood was collected from the posterior ocular venous plexus, with approximately 0.20 mL collected per sample. The blood samples were anticoagulated with EDTA and placed on ice after collection. The plasma was separated by centrifugation within 2 hours (centrifugation conditions: 3500 rpm, 10 minutes, 2-8°C).

[0049] The HPLC-MS conditions are shown in Table 5.

[0050] Table 5

[0051] Continued table

[0052] 2.3. Results The main pharmacokinetic parameters of each formulation are shown in Table 6.

[0053] Table 6 F4 1.0±0.0 89±153 869±1564 2.68 F5 1.3±0.5 170±230 1817±2727 5.60 F6 1.8±0.5 124±73.3 1296±838 3.99 F7 1.5±0.6 96.4±45.3 915±532 2.82 F8 1.0±0.0 194±48 1996±887 6.15 F9 2.0±0.0 45.5±89 469±936 1.44 F10 1.3±0.5 117±62.9 1076±571 3.31 F11 0.81±0.239 23.76±21.1 141±137 0.22 F12 0.54±0.529 14.98±5.18 72.60±43.0 0.11 F13 0.40±0.21 13.06±14.9 73.49±113 0.05 F14 1.00±0.00 11.52±3.63 59.53±25.18 0.04 F15 1.02±0.783 105±140 722±1022 1.11 F16 0.88±0.144 118±108 915±973 1.41 F18 0.94±0.13 23.51±14.78 112±78.90 0.34 F19 1.00±0.00 105.48±92 735±676 2.26 F20 0.94±0.13 73.90±33.3 527±319 1.62 F21 7.00±1.15 110±24.6 1926±375 N / A F22 4.00±1.63 157±30.8 2783±676 N / A F23 6.75±4.57 323±92.2 5804±1289 N / A *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 injection dosage; AUCR: Area under the subcutaneous injection blood drug concentration-time curve; DT: Dosage of nasal drops The results of the above animal experiments show that different concentrations of dodecyl-β-D-maltodextrin (DDM) and polyoxyethylene (9) dodecyl ether (C12E9) in different ratios with smegglutinin can be well absorbed into the blood.

[0054] Example 3. Compatibility / Stability Study The changes in semaglutide content and impurities in semaglutide solution, semaglutide plus different penetration enhancers solution, and semaglutide plus different antibacterial agents solution were determined by HPLC under the conditions of Example 1 and stored in the dark at 40℃ and 50℃ for 10 days (10D). The results are shown in Table 7.

[0055] Table 7

[0056] Continued table

[0057] Note: SEM is semaglutide. All formulations are aqueous solutions, and the concentration of SEM is 5.252 mg / ml.

[0058] The definitions of hydrophilic impurities 1 and 2 and hydrophobic impurities mentioned in Table 7 are as follows: Figures 1-4 The HPLC chromatograms of prescriptions C1-C4 shown are obtained after 10 days at 50°C.

[0059] As a control, the main peak intensities of compositions J and K of CN116942612A at 50°C for 5 days were determined using the same method, and both were below 95%.

[0060] Results: Both penetration enhancers protected semaglutide and inhibited its degradation; benzalkonium chloride, as an antibacterial agent in nasal spray formulations, also protected semaglutide in this product.

[0061] Example 4. Pharmacokinetic Study - Brain Tissue Distribution 4.1. Test Materials and Test Substances (1) Animals: Same as in Example 2. 40 rats in each test prescription group.

[0062] (2) Test formulations: F5, F8 and F23 in Example 2.

[0063] 4.2. Methods Administration: Same as in Example 2.

[0064] sampling: Five male and five female rats were euthanized by dislocation at 1, 8, 16, and 24 hours after drug administration. Brains were collected, quickly rinsed with water, accurately weighed, and then minced. The tissue was mixed with pre-cooled physiological saline at a ratio of 1 g: 3 mL, homogenized using an ultrasonic cell homogenizer, and centrifuged at 12000 rpm for 2 min. The supernatant was collected and stored at -20℃ for later analysis. The concentration of smegglutinin 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.

[0065] Testing: The conditions are the same as in Example 2.

[0066] 4.3. Results As shown in Table 8.

[0067] Table 8

[0068] BLQ = below the detection limit As shown in Table 8, the brain distribution of rats after intranasal administration of F5 and F8 described in this invention was significantly lower than that after injection administration of F23.

[0069] Example 5. Pharmacokinetics – Brain Tissue Distribution The method is the same as in Example 4, except that the types of test prescriptions are increased, namely, prescriptions selected from F6, F7, F9, F10, F13 and F14 in Table 6 are tested.

[0070] Prescriptions F6, F7, F9, and F10 showed detectable semaglutide in brain tissue at at least one time point; among them, the semaglutide dosage of F6, F7, and F10 was the same as that of F5 and F8, but the types of nasal absorption enhancers were different, indicating that there is a great deal of unpredictability in whether nasal absorption enhancers can avoid brain delivery.

[0071] In addition, no brain drug distribution was detected at any time point for the highest doses F13 and F14.

Claims

1. A composition containing a GLP-1 receptor agonist and a nasal absorption enhancer, characterized in that, The GLP-1 receptor agonist is smegglutinin; the nasal absorption enhancer is selected from dodecyl-β-D-maltodextrin or polyoxyethylene (9) dodecyl ether; wherein the mass ratio of the nasal absorption enhancer to the GLP-1 receptor agonist in the composition is 1:1.

05.

2. The composition according to claim 1, characterized in that, The composition contains a nasal absorption enhancer with a w / v% content of 0.5% and a GLP-1 receptor agonist with a content of 5.25 mg / mL.

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

4. The composition according to claim 1 or 3, characterized in that, The composition uses water as a solvent.

5. The composition according to claim 1 or 3, characterized in that, The composition further contains a preservative.

6. The composition according to claim 5, characterized in that, The preservative is phenol, benzalkonium chloride, benzyl ammonium chloride, 2-trichloromethyl-2-propanol, butyl p-hydroxybenzoate, propyl p-hydroxybenzoate, benzyl chloride, chlorocresol, or benzoic acid.

7. The composition according to claim 5, characterized in that, The preservative is benzalkonium chloride.

8. Use of the composition according to any one of claims 1-7 in the preparation of a medicament for treating diseases, characterized in that, The disease mentioned is selected from one or more of type 2 diabetes, overweight, or obesity.

9. A method for preparing the composition according to any one of claims 1-7, characterized in that, This includes mixing GLP-1 receptor agonists with nasal absorption enhancers.

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