Bremelanotide-containing pharmaceutical composition, formulation thereof, preparation method therefor, and use thereof

Abstract

A bremelanotide pharmaceutical composition, a formulation thereof, a preparation method therefor, and use thereof. The bremelanotide pharmaceutical composition comprises bremelanotide as a pharmaceutically active ingredient, a solvent, and a polymer. The solvent includes one or more of N-methylpyrrolidone, polyethylene glycol, dimethyl sulfoxide, and benzyl benzoate and enables the plasma concentration of bremelanotide to be stably greater than 0.1 ng / mL at 120-168 h, thereby achieving the target sustained release. The bremelanotide pharmaceutical composition can be directly injected and does not require the addition of a thickener, a gel-forming agent, or other excipients.

Description

A pharmaceutical composition containing Bremerlongdan, a formulation thereof, a method for its preparation and its application Technical Field

[0001] This invention belongs to the field of pharmaceutical formulation technology, specifically relating to a pharmaceutical composition containing Bremerlongdan, a formulation thereof, its preparation method and application. Background Technology

[0002] Bremerlantoide, also known as Bremerlantoide PT141, Bremerlantoide impurity, Bremerlantoide, and Bremerlantoide, is a cyclic heptapeptide compound with the molecular formula C. 50 H 68 N 14 O 10 Its chemical structural formula is shown in Formula 1:

[0003] Bremerlongtan, as a ligand for one or more melanocortin receptors, can bind well to melanocortin receptors and produce good effects. Its pharmacological characteristics are that the drug has the potential to treat or prevent acute or chronic inflammatory diseases, autoimmune diseases, transplant rejection, metabolic diseases with weight gain, metabolic diseases with weight loss, diabetes, diabetic complications, cancer, cancerous hyperplasia, reproductive system diseases, peripheral or central nervous system diseases, cardiovascular diseases, or respiratory system diseases.

[0004] Chinese invention patent application 202111481324.8 discloses a method for synthesizing Bremerlongan acetate, specifically: using the Fmoc solid-phase polypeptide synthesis method, with RinkAmide resin as a carrier, after modifying it with 4-trifluoromethylmandelic acid, according to the polypeptide sequence of Bremerlongan, amino acids with Fmoc-protecting groups are sequentially condensed from the C-terminus to the N-terminus under the action of a coupling agent to obtain a linear peptide resin; then, the Fmoc protecting group and side chain protecting agents such as Lys and Asp are sequentially removed; then, cyclization is formed on the resin under the action of a coupling agent; the resin is cleaved with a cleavage agent; the ester bond is broken under alkaline conditions to obtain crude Bemenotide peptide; finally, after purification, refined Bemenotide peptide is obtained. The method is simple, has a high yield, and high purification efficiency.

[0005] Research on the original Bremerlongdan drug-device combination on the market revealed that VYLEESI (AUTOINJECTOR), the original Bremerlongdan product, as a subcutaneous administration drug, achieves peak plasma concentration rapidly in 0.5-1.0 hours, has 100% bioavailability, a half-life of 1.9-4.0 hours, and an average clearance of 6.5 ± 1.0 L / hr. However, it has limitations in indications requiring long-term dosing. Furthermore, existing sustained-release formulations on the market often require mixing before use, which introduces additional inconvenience and potential medication errors for patients.

[0006] Given the limitations of existing technologies, there is an urgent need in the field to provide a long-acting sustained-release formulation of Bremerlongdan that can meet the needs of long-term administration through continuous release technology. Summary of the Invention

[0007] This invention addresses the problems existing in the prior art by providing a pharmaceutical composition containing Bremerlongdan, a formulation thereof, a method for its preparation, and its application.

[0008] To achieve the above objectives, the technical solution adopted by the present invention is as follows:

[0009] A pharmaceutical composition comprising Bremerlongdan, a solvent, and a polymer, said solvent comprising one or more of N-methylpyrrolidone, polyethylene glycol, dimethyl sulfoxide, and benzyl benzoate.

[0010] Preferably, the solvent is a mixed solution of dimethyl sulfoxide, N-methylpyrrolidone and benzyl benzoate, a mixed solution of dimethyl sulfoxide and benzyl benzoate, or a mixed solution of N-methylpyrrolidone and dimethyl sulfoxide.

[0011] More preferably, the solvent is a mixed solution of N-methylpyrrolidone and benzyl benzoate, wherein N-methylpyrrolidone accounts for 5-95% of the volume of the mixed solution.

[0012] More preferably, the solvent is a mixed solution of N-methylpyrrolidone and benzyl benzoate, wherein N-methylpyrrolidone accounts for 95% of the volume of the mixed solution.

[0013] More preferably, the solvent is a mixed solution of dimethyl sulfoxide and benzyl benzoate, wherein dimethyl sulfoxide accounts for 5-95% of the volume of the mixed solution.

[0014] More preferably, the solvent is a mixed solution of dimethyl sulfoxide and benzyl benzoate, wherein dimethyl sulfoxide accounts for 95% of the volume of the mixed solution.

[0015] More preferably, the solvent is a mixed solution of N-methylpyrrolidone and dimethyl sulfoxide, wherein N-methylpyrrolidone accounts for 5-95% of the volume of the mixed solution.

[0016] Preferably, the polymer is a copolymer of lactide and glycolide, carbomer, or sodium alginate.

[0017] More preferably, the polymer is a lactide-glycolic acid copolymer.

[0018] Preferably, the end-capping structure of the lactide-glycol copolymer is carboxyl-terminated or ester-terminated.

[0019] Preferably, the molar ratio of lactic acid monomer to glycolic acid monomer in the lactide-glycolic acid copolymer is 50-85:15-50.

[0020] More preferably, the molar ratio of lactic acid monomer to glycolic acid monomer in the lactide-glycolic acid copolymer is 50-75:20-50.

[0021] More preferably, the molar ratio of lactic acid monomer to glycolic acid monomer in the lactide-glycolic acid copolymer is 50:50 or 75:25.

[0022] Preferably, the average molecular weight of the lactide-glycolic acid copolymer is 20-50 kDa.

[0023] More preferably, the average molecular weight of the lactide-glycolic acid copolymer is 20-25 kDa.

[0024] More preferably, the average molecular weight of the lactide-glycolic acid copolymer is 23 kDa.

[0025] Preferably, the pharmaceutical composition comprises, by weight percentage, 0.1-15% Bremerlongtan, 10-70% polymer, and solvent to bring the total to 100%.

[0026] More preferably, the pharmaceutical composition comprises, by weight percentage, 1-5% Bremerlongtan, 30-50% polymer, and solvent to bring the total to 100%.

[0027] More preferably, the pharmaceutical composition comprises, by weight percentage, 1-1.5% Bremerlongtan, 35-40% polymer, and solvent to make up to 100%.

[0028] The present invention also provides a method for preparing the above-mentioned pharmaceutical composition, comprising directly mixing Bremerlongdan, solvent and polymer to obtain the composition.

[0029] The present invention also provides a pharmaceutical formulation comprising the above-described pharmaceutical composition or a pharmaceutical composition prepared by the above-described preparation method.

[0030] Preferably, the preparation is an injectable formulation.

[0031] The present invention also provides the use of the above-described pharmaceutical composition or the pharmaceutical composition or pharmaceutical preparation obtained by the above-described preparation method in the preparation of sustained-release Bremerlongdan.

[0032] Compared with the prior art, the present invention has the following beneficial effects:

[0033] (1) The pharmaceutical composition containing Bremerlongdan provided by the present invention enables the blood concentration of Bremerlongdan to remain stable at greater than 0.1 ng / mL for 120-168 h, thereby achieving the target sustained release.

[0034] (2) The drug composition containing Bremerlongdan prepared in this invention can be injected directly without mixing the drug solution before use.

[0035] (3) The pharmaceutical composition containing Bremerlongdan provided by the present invention has a simple preparation process, a single component, and can be mass-produced without the need to add thickeners, gelling agents or other excipients. It has low cost and good market application prospects. Detailed Implementation

[0036] The present invention will be further described below with reference to specific embodiments, and the advantages and features of the present invention will become clearer as a result of the description. However, these examples are merely exemplary and do not constitute any limitation on the scope of the present invention. Unless otherwise specified, the methods, ingredients, and dosages involved in the following embodiments are conventional methods known to those skilled in the art.

[0037] It is worth noting that the raw materials used in this invention are all commercially available products, and the sources of some raw materials are shown in Table 1.

[0038] Table 1 Raw Material Source Information

[0039] 1. The abbreviations used in this invention have the following meanings:

[0040] API: Key pharmaceutical ingredient;

[0041] NMP: N-methyl-2-pyrrolidone;

[0042] DMSO: Dimethyl sulfoxide;

[0043] PLGA: Lactide-glycolic acid copolymer;

[0044] PBS: Phosphate Buffered Salt;

[0045] BA: Benzyl alcohol;

[0046] BB: Benzyl benzoate.

[0047] 2. The specific method for the in vitro release experiment of the present invention is as follows:

[0048] Dissolution method: Add an appropriate amount of the prepared drug gel, weighing about 20 mg, dropwise into the medium and let it stand in a water bath at 37°C;

[0049] Dissolution medium: PBS at pH 7.4 and a concentration of 20 mg / mL;

[0050] Medium volume: 10 mL;

[0051] Sampling method: Take 1 mL each time, and then add 1 mL of fresh medium;

[0052] Sampling points: Samples were taken at 0.5h and 24h to determine the content of API Bremerlongtan.

[0053] 3. The API content of this invention is detected by HPLC, and the specific method is as follows:

[0054] 3.1 Solution Preparation

[0055] 3.1.1 Diluent: Weigh about 1g of bovine serum albumin into a blue-capped bottle, add 800mL of purified water to dissolve it, add 200mL of acetonitrile, add 1mL of trifluoroacetic acid, and shake well to obtain the product.

[0056] 3.1.2 Preparation of mobile phase: Measure 700 mL of purified water into a blue-capped bottle, add 300 mL of acetonitrile, mix well, add 1 mL of trifluoroacetic acid, mix well, filter through a 0.45 μm filter membrane, and degas by sonication to obtain the mobile phase.

[0057] 3.1.3 Preparation of control solution: Accurately weigh about 10 mg of API Bremerlongtan into a 20 mL volumetric flask, add diluent to dissolve and dilute to the mark, and shake well; measure 1.0 mL of the above solution into a 20 mL volumetric flask, add diluent to dilute to the mark, and shake well to obtain the control solution.

[0058] 3.1.4 Preparation of test solution: Weigh approximately 50 mg of the test sample accurately into a 20 mL volumetric flask, add 4 mL of DMSO and sonicate to dissolve. Add diluent and dilute to the mark, shake well, take an appropriate amount into a centrifuge tube, centrifuge at 8000 rpm for 10 min, and take the supernatant.

[0059] 3.2. Chromatographic conditions

[0060] Instrument: High-performance liquid chromatograph;

[0061] Column: Waters Xbridge C18, 150mm × 4.6mm × 3.5μm;

[0062] Mobile phase: Acetonitrile:water = 30:70 (v:v, containing 0.1% trifluoroacetic acid);

[0063] Column temperature: 35℃;

[0064] Flow rate: 1.0 mL / min;

[0065] Detection wavelength: 215nm;

[0066] Injection volume: 20 μL;

[0067] Running time: 10 minutes;

[0068] Elution method: isocratic elution.

[0069] Example 1

[0070] The composition of a pharmaceutical composition containing Bremerlongdan is shown in Table 2 below.

[0071] Table 2 Prescriptions

[0072] A method for preparing a pharmaceutical composition containing Bremerlongtan: accurately weigh the prescribed amounts of Bremerlongtan, PLGA and DMSO into a sample vial, then place the sample vial in a roller mixer and roll it until the Bremerlongtan and PLGA are completely dissolved.

[0073] Example 2

[0074] The composition of a pharmaceutical composition containing Bremerlongdan is shown in Table 3 below.

[0075] Table 3 Prescriptions

[0076] A method for preparing a pharmaceutical composition containing Bremerlongtan: accurately weigh the prescribed amounts of Bremerlongtan, PLGA, DMSO and BB into a sample vial, and then place the sample vial in a roller mixer until Bremerlongtan and PLGA are completely dissolved.

[0077] Experiment Example 1: In Vitro Release Experiment

[0078] 1. Solubility of API Bremerlongtan

[0079] To select a suitable solvent while minimizing the amount of API used while meeting the drug concentration requirements in the formulation, the concentration of the drug Bremerlongan was set at 40 mg / mL. The saturated solubility (mg / mL) of Bremerlongan in 5 mL of different aqueous or organic solvents was investigated. The aqueous solvent was water or PBS with a pH of 7.4 and a concentration of 20 mg / mL. The organic solvents were NMP, DMSO, a mixture of NMP and BB at a volume ratio of 1:0.05, or a mixture of NMP and BA at a volume ratio of 1:0.05. The test results are shown in Table 4.

[0080] As shown in Table 4, the saturation solubility of Bremerlongtan in PBS at pH 7.4 is relatively low.

[0081] Table 4. Saturated solubility results of API in different solvents

[0082] 2. Prescription screening

[0083] Because of the good water solubility of API Bremerlongtan, it is difficult to develop sustained-release formulations. Therefore, during the development process, we prioritized exploring the feasibility of extreme formulations, that is, using in vitro release rate as the evaluation index, and focusing on screening the average molecular weight of PLGA, the end-capped structure of PLGA, the molar ratio of lactic acid monomer and glycolic acid monomer in PLGA, and the solvent system.

[0084] 2.1 Average molecular weight of PLGA

[0085] PLGA with carboxyl-terminated, lactic acid, and glycolic acid monomers in a molar ratio of 50:50 and different average molecular weights (20, 23, 25, and 40 kDa) were selected, and the solvents were NMP or DMSO.

[0086] By weight percentage, 1.1% of Bremerlongtan, 36% of PLGA and 62.9% of solvent were mixed in a roller mixer to prepare a pharmaceutical composition containing Bremerlongtan. The molecular weight of PLGA was screened using the in vitro release rate as the evaluation index. The results are shown in Table 5.

[0087] As shown in Table 5, the in vitro release rates of the drug compositions prepared from PLGA with average molecular weights of 20, 23, and 25 kDa are low. The solvents and the combinations of average molecular weights of PLGA shown can all be used in the formulation of the Bremerlongdan composition.

[0088] Table 5. Results of in vitro release experiments of drug compositions prepared with different average molecular weights of PLGA

[0089] 2.2 PLGA End Cap Structure

[0090] PLGA with a molar ratio of lactic acid monomer to glycolic acid monomer of 50:50, an average molecular weight of 40 kDa, and different end-capping structures (carboxyl-capped and ester-capped) was selected, and NMP was used as the solvent.

[0091] By weight percentage, 1.1% of Bremerlongtan, 36% of PLGA and 62.9% of solvent were mixed in a roller mixer to prepare a pharmaceutical composition containing Bremerlongtan. The in vitro release rate was used as the evaluation index to screen the end-capped structure of PLGA. The results are shown in Table 6.

[0092] As shown in Table 6, both carboxyl-terminated and ester-terminated PLGA can be used in the formulation of Bremerlongdan compositions. However, the drug composition prepared from ester-terminated PLGA has a lower in vitro release rate.

[0093] Table 6. In vitro release test results of drug compositions prepared with different PLGA end-capped structures

[0094] 2.3 Molar ratio of lactic acid monomer to glycolic acid monomer in PLGA

[0095] PLGA with carboxyl-terminated groups, an average molecular weight of 23 kDa, and different molar ratios of lactic acid monomers and glycolic acid monomers were selected, and NMP was used as the solvent.

[0096] By weight percentage, 1.2% of Bremerlongtan, 39.5% of PLGA and 59.3% of solvent were mixed in a roller mixer to prepare a pharmaceutical composition containing Bremerlongtan. The molar ratio of lactic acid monomer and glycolic acid monomer in PLGA was screened using the in vitro release rate as the evaluation index. The results are shown in Table 7.

[0097] As shown in Table 7, PLGA with a molar ratio of lactic acid monomer to glycolic acid monomer of 75:25 or 50:50 can be used in the formulation of Bremerlongdan compositions. Among them, the drug composition prepared from PLGA with a molar ratio of lactic acid monomer to glycolic acid monomer of 50:50 has a lower in vitro release rate.

[0098] Table 7. In vitro release test results of drug compositions prepared from PLGA with different molar ratios of lactic acid monomer and glycolic acid monomer.

[0099] 2.4 Solvent System

[0100] PLGA with carboxyl-terminated ends, an average molecular weight of 23 kDa, and a molar ratio of lactic acid monomer to glycolic acid monomer of 50:50 was selected. The solvents were NMP, DMSO, a mixed solution of NMP and BB with a volume ratio of 0.95:0.05, and a mixed solution of DMSO and BB with a volume ratio of 0.95:0.05.

[0101] By weight percentage, the NMP system contains 1.2% Bremerlongtan, 39.5% PLGA, and 59.3% solvent; the DMSO system contains 1.4% Bremerlongtan, 39.4% PLGA, and 59.2% solvent. The above materials are mixed in a roller mixer to prepare a pharmaceutical composition containing Bremerlongtan.

[0102] Solvents were screened based on in vitro release rate, and the results are shown in Table 8.

[0103] As can be seen from Table 8, the solvents in the table can all be used in the formulation of Bremelanotide compositions. Among them, the in vitro release rate of the pharmaceutical compositions prepared from a mixed solution of DMSO, NMP and BB with a volume ratio of 0.95:0.05 or a mixed solution of DMSO and BB with a volume ratio of 0.95:0.05 is relatively low.

[0104] In vitro release experiment results of pharmaceutical compositions prepared with different solvents in Table 8

[0105] 3. Prescription design

[0106] Combined with the above PLGA and solvent screening, 4 prescriptions with relatively low in vitro release rates were obtained. The specific information is shown in Table 9. Two of the prescriptions were selected as Example 1 and Example 2.

[0107] Table 9 Prescriptions of pharmaceutical compositions containing Bremelanotide

[0108] Experimental Example 2 Pharmacokinetics (PK) experiment in rats

[0109] Experimental animals: 10 SD rats, SPF grade males, weighing 210 - 240 g, purchased from Guangzhou Ruige Biotechnology Co., Ltd., production license number SYXK (Guangdong) 2022 - 0296. Adapted to the environment for 3 - 5 days, with free access to food and water. The feed was Co 60 Irradiated experimental rat maintenance feed. The environment in the animal house was maintained at a temperature of 20 - 26 °C, humidity of 40 - 70%, 12 - hour light - dark cycle, ventilation of 10 - 20 times / h, and no more than 3 rats were housed in each cage.

[0110] Experimental method: 10 SD rats were randomly divided into 3 groups. Group 1 (n = 4) was subcutaneously administered with 3 mg / kg of the Bremelanotide - containing pharmaceutical composition prepared in Example 1; Group 2 (n = 3) was subcutaneously administered with 6 mg / kg of the Bremelanotide - containing pharmaceutical composition prepared in Example 1; Group 3 (n = 3) was subcutaneously administered with 6 mg / kg of the Bremelanotide - containing pharmaceutical composition prepared in Example 2. Pharmacokinetic blood samples were collected and serum was separated at 0 h (before dosing) and 0.5, 1, 1.5, 2, 3, 4, 6, 12, 24, 48, 72, 96, 120 and 168 h after dosing to detect plasma drug concentration.

[0111] Experimental Results and Analysis: The experimental results are shown in Table 10. Seven days (168 hours) after administration, no obvious abnormalities were found at the administration sites in all three groups of animals. The PK test results showed that the drug was released 24 hours after administration. In Groups 1 and 2, with administration doses of 3 mg / kg and 6 mg / kg respectively, the plasma drug concentration at 120 hours after administration was higher than 0.1 ng / mL. In Groups 2 and 3, with an administration dose of 6 mg / kg, the plasma drug concentration at 168 hours after administration was also higher than 0.1 ng / mL.

[0112] Table 10 Results of the rat PK experiment Note: BQL indicates below the lower limit of quantitation.

[0113] The pharmacokinetic properties of the pharmaceutical compositions containing Bremerlongdan prepared in Examples 1-2 of this invention were further investigated.

[0114] The pharmacokinetic data of Examples 1-2 were compared with the pharmacokinetic data of the original product VYLEESI (AUTOINJECTOR) in the FDA review report. The pharmacokinetic data are shown in Table 11.

[0115] VYLEESI: Each pre-filled syringe contains 1.75 mg bremeropeptide (equivalent to 1.89 mg bremeropeptide acetate) in 0.3 mL of solution. Inactive ingredients include 2.5% glycerol and sterile water for injection. Hydrochloric acid or sodium hydroxide is used to adjust the pH.

[0116] As shown in Table 11, the pharmaceutical compositions containing Bramerlongtan prepared in Examples 1-2 all exhibited good sustained-release effects and effectively reduced C. max , will T max It shifted backward and greatly prolonged T. 1 / 2 This effectively achieves the goal of slow release.

[0117] Table 11 Pharmacokinetic Data Note: * This indicates that the VYLEESI data comes from the FDA review report of the original product.

[0118] Finally, it should be noted that the above content is only used to illustrate the technical solution of the present invention, and is not intended to limit the scope of protection of the present invention. Simple modifications or equivalent substitutions made by those skilled in the art to the technical solution of the present invention do not depart from the essence and scope of the technical solution of the present invention.

Claims

1. A pharmaceutical composition, characterized in that, It includes the pharmaceutically active ingredient Bremerlongtan, a solvent, and a polymer, wherein the solvent includes one or more of N-methylpyrrolidone, polyethylene glycol, dimethyl sulfoxide, and benzyl benzoate.

2. The pharmaceutical composition according to claim 1, characterized in that, The solvent is a mixed solution of dimethyl sulfoxide, N-methylpyrrolidone and benzyl benzoate, a mixed solution of dimethyl sulfoxide and benzyl benzoate, or a mixed solution of N-methylpyrrolidone and dimethyl sulfoxide.

3. The pharmaceutical composition according to claim 1, characterized in that, The polymer is a copolymer of lactide and glycolide, carbomer, or sodium alginate.

4. The pharmaceutical composition according to claim 3, characterized in that, The polymer is a lactide-glycolic acid copolymer, and the end-capping structure of the lactide-glycolic acid copolymer is carboxyl-terminated or ester-terminated.

5. The pharmaceutical composition according to claim 4, characterized in that, The molar ratio of lactic acid monomer to glycolic acid monomer in the lactide-glycolic acid copolymer is 50-85:15-50.

6. The pharmaceutical composition according to claim 4, characterized in that, The average molecular weight of the lactide-glycolic acid copolymer is 20-50 kDa.

7. The pharmaceutical composition according to any one of claims 1-6, characterized in that, The pharmaceutical composition comprises, by weight percentage, 0.1-15% Bremerlongtan, 10-70% polymer, and solvent to bring the total to 100%.

8. A method for preparing a pharmaceutical composition according to any one of claims 1-7, characterized in that, This involves directly mixing the active pharmaceutical ingredient, Bremerlongtan, solvent, and polymer to obtain the final product.

9. A pharmaceutical preparation, characterized in that, The pharmaceutical composition includes the pharmaceutical composition according to any one of claims 1-7 or the pharmaceutical composition prepared by the preparation method according to claim 8, wherein the pharmaceutical preparation is of the type of injection.

10. The use of a pharmaceutical composition according to any one of claims 1-7, a pharmaceutical composition prepared by the preparation method according to claim 8, or a pharmaceutical formulation according to claim 9 in the preparation of a sustained-release Bramer-Landan drug.

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